21,314 results
Search Results
2. Spheroids formation in large drops suspended in superhydrophobic paper cones.
- Author
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Mohapatra, Omkar, Gopu, Maheshwar, Ashraf, Rahail, Easo George, Jijo, Patil, Saniya, Mukherjee, Raju, Kumar, Sanjay, and Mampallil, Dileep
- Subjects
- *
SUPERHYDROPHOBIC surfaces , *CELL culture , *MEDICAL screening , *DRUG development , *OVARIAN cancer , *CANCER cells - Abstract
The utilization of 3D cell culture for spheroid formation holds significant implications in cancer research, contributing to a fundamental understanding of the disease and aiding drug development. Conventional methods such as the hanging drop technique and other alternatives encounter limitations due to smaller drop volumes, leading to nutrient starvation and restricted culture duration. In this study, we present a straightforward approach to creating superhydrophobic paper cones capable of accommodating large volumes of culture media drops. These paper cones have sterility, autoclavability, and bacterial repellent properties. Leveraging these attributes, we successfully generate large spheroids of ovarian cancer cells and, as a proof of concept, conduct drug screening to assess the impact of carboplatin. Thus, our method enables the preparation of flexible superhydrophobic surfaces for laboratory applications in an expeditious manner, exemplified here through spheroid formation and drug screening demonstrations. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
3. High sensitivity and automatic chemiluminescence detection of glucose and lactate using a spin-disc paper-based device.
- Author
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Tong, Wenqiang, Shi, Jiaming, Yu, Zhihang, Ran, Bin, Chen, Huaying, and Zhu, Yonggang
- Subjects
- *
CHEMILUMINESCENCE , *LACTATES , *ARTIFICIAL saliva , *LACTATION , *GLUCOSE , *SPIN labels , *SIGNAL detection , *CELL culture , *MONOCARBOXYLATE transporters - Abstract
This paper reports a spin-disc paper-based device with 10 individual detection units containing electromagnetic modules controlling the sample incubation time before chemiluminescence (CL) signal detection. After the sample was added to the top paper chip and incubated with the enzyme, the electromagnet was turned off to allow contact between the top and bottom paper. The H2O2 generated by the sample flowed vertically to the bottom paper and initiated the oxidase of the luminol to generate the CL signal. After one detection the disc was automatically rotated to the next position to repeat the above detection. The advantage of using the device over the lateral flow and the in situ detection was firstly proved using the detection of H2O2 and the glucose/lactate sample with 5 minute incubation. The CL intensity was increased 300 times/1000 times as the glucose/lactate was incubated for 5 minutes compared to the non-incubated samples. Afterward, the device was employed to separately detect glucose and lactate diluted in PBS, artificial sweat, artificial saliva, and fresh cell culture media. Finally, the device was employed to detect the glucose and lactate in the media collected over the 24 hour culture of PC3 cells. The uptake and production rates of glucose and lactate were correspondingly determined as 0.328 ± 0.015 pmol h−1 per cell and 1.254 ± 0.053 pmol h−1 per cell, respectively. The reported device has wide application potential due to its capabilities in automatic detection of multiple samples with very high sensitivity and small sample volume (down to 0.5 μL). [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
4. Paper-based plasmonic substrates as surface-enhanced Raman scattering spectroscopy platforms for cell culture applications
- Author
-
J.M. Romo-Herrera, K. Juarez-Moreno, L. Guerrini, Y. Kang, N. Feliu, W.J. Parak, and R.A. Alvarez-Puebla
- Subjects
Plasmonic nanoparticles ,Paper-based substrates ,SERS ,Cell culture ,Plasmonic papers ,Medicine (General) ,R5-920 ,Biology (General) ,QH301-705.5 - Abstract
The engineering of advanced materials capable of mimicking the cellular micro-environment while providing cells with physicochemical cues is central for cell culture applications. In this regard, paper meets key requirements in terms of biocompatibility, hydrophilicity, porosity, mechanical strength, ease of physicochemical modifications, cost, and ease of large-scale production, to be used as a scaffold material for biomedical applications. Most notably, paper has demonstrated the potential to become an attractive alternative to conventional biomaterials for creating two-dimensional (2D) and three-dimensional (3D) biomimetic cell culture models that mimic the features of in vivo tissue environments for improving our understanding of cell behavior (e.g. growth, cell migration, proliferation, differentiation and tumor metastasis) in their natural state. On the other hand, integration of plasmonic nanomaterials (e.g. gold nanoparticles) within the fibrous structure of paper opens the possibility to generate multifunctional scaffolds equipped with biosensing tools for monitoring different cell cues through physicochemical signals. Among different plasmonic based detection techniques, surface-enhanced Raman scattering (SERS) spectroscopy emerged as a highly specific and sensitive optical tool for its extraordinary sensitivity and the ability for multidimensional and accurate molecular identification. Thus, paper-based plasmonic substrates in combination with SERS optical detection represent a powerful future platform for monitoring cell cues during cell culture processes. To this end, in this review, we will describe the different methods for fabricating hybrid paper-plasmonic nanoparticle substrates and their use in combination with SERS spectroscopy for biosensing and, more specifically, in cell culture applications.
- Published
- 2021
- Full Text
- View/download PDF
5. Paper-based cell culture microfluidic system
- Author
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Tao, Fang Fang, Xiao, Xia, Lei, Kin Fong, and Lee, I-Chi
- Published
- 2015
- Full Text
- View/download PDF
6. Selecting the appropriate indirect viability assay for 3D paper-based cultures: a data-driven study.
- Author
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Sitte, Zachary R., Larson, Tyler S., McIntosh, Julie C., Sinanian, Melanie, and Lockett, Matthew R.
- Subjects
- *
FLUORESCENT proteins , *PROPIDIUM iodide , *ANTINEOPLASTIC agents , *RESEARCH questions , *PROTEIN expression , *QUINAZOLINONES , *CELL culture - Abstract
Cellular viability measurements quantify decreased proliferation or increased cytotoxicity caused by drug candidates or potential environmental toxins. Direct viability measures count each cell to provide an accurate readout. This approach can prove analytically challenging and time-consuming when cells are maintained in 3D structures akin to tissues or solid tumors. While less labor-intensive, indirect viability measures can be less accurate due to the heterogeneous structural and chemical microenvironment that arises when cells are maintained in tissue-like architectures and in contact with extracellular matrices. Here we determine the analytical figures of merit of five indirect viability assays in the paper-based cell culture platform we continue to develop in our laboratory: calcein-AM staining, the CellTiter-Glo assay, imaging fluorescent protein expression, propidium iodide staining, and the resazurin assay. We also determined the compatibility of each indirect assay with hypoxic conditions, intra-experimental repeatability, inter-experimental reproducibility, and ability to predict a potency value for a known antineoplastic drug. Our results show that each assay has benefits and drawbacks to consider when choosing the appropriate readout to answer a particular research question. We also highlight that only one indirect readout is unaffected by hypoxia, a commonly overlooked variable in cell culture that likely yields inaccurate viability measures. [ABSTRACT FROM AUTHOR]
- Published
- 2023
- Full Text
- View/download PDF
7. Anionic Polysaccharide-Modified Filter Papers for Rapid Isolation of Extracellular Vesicles from Diverse Samples in a Simple Bind–Wash–Elute Manner
- Author
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Kaiyi Li, Qiang Liu, Zhi Geng, Shanglin Li, Peng Liu, and Tian Zhao
- Subjects
Lung Neoplasms ,Chromatography ,Filter paper ,Elution ,010401 analytical chemistry ,RNA ,010402 general chemistry ,01 natural sciences ,0104 chemical sciences ,Analytical Chemistry ,ErbB Receptors ,Extracellular Vesicles ,chemistry.chemical_compound ,chemistry ,Polysaccharides ,Cell culture ,Spin column-based nucleic acid purification ,Mutation ,Nucleic acid ,Humans ,Ultracentrifuge ,Protein Kinase Inhibitors ,DNA - Abstract
Extracellular vesicles (EVs) play a significant role in the pathophysiological process of many diseases, highlighting their values in medical diagnosis and disease monitoring. However, the current EV isolation methods are time-consuming, inconvenient to operate, and incompatible with downstream analyses. Here, we present a novel isolation method employing anionic polysaccharide-modified filter papers for the isolation of EVs (AppiEV) via electrostatic adsorption. A disc of glass fiber-based filter modified with sodium alginate was assembled into a spin column to function as the solid capture phase. In the acidic condition, EVs were induced to carry more positively charged ions, which enable the capture of EVs by the negatively charged filter paper. After a wash, the EVs were released from the spin column using an alkaline elution buffer, which induces the EVs to carry more negative charges. The EVs isolated by AppiEV from cell culture supernatants, plasma, and urine are similar to or even better than those isolated by ultracentrifugation in terms of EV size distribution, protein distribution, and nucleic acid contents. Due to the interference removal of the EV-free RNA and DNA attributed to the negatively charged capture medium, the eluate of AppiEV could be directly used for genetic analysis, including the stem-loop RT-PCR analysis of miR-21 and the allele-specific PCR analysis of mutation genes of EGFR p.L858R and EGFR p.T790M. We believe that AppiEV offers a simple and efficient approach for the isolation of high-quality EVs from various liquid specimens.
- Published
- 2021
8. Paper based microfluidic platform for single-step detection of mesenchymal stromal cells secreted VEGF
- Author
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Enrique Azuaje-Hualde, Marian M. de Pancorbo, Fernando Benito-Lopez, Lourdes Basabe-Desmonts, and European Commission
- Subjects
Paper ,Vascular Endothelial Growth Factor A ,structure switching signaling aptamer ,cell culture ,vascular endothelial growth factor ,Vascular Endothelial Growth Factors ,Microfluidics ,paper microfluidic device ,Mesenchymal Stem Cells ,Biosensing Techniques ,Microfluidic Analytical Techniques ,cell secretion ,Biochemistry ,Analytical Chemistry ,Environmental Chemistry ,Spectroscopy - Abstract
[EN] Low cost and user-friendly paper microfluidic devices, combined with DNA-based biosensors with binding capacities for specific molecules, have been proposed for the developing of novel platforms that ease and speed-up the process of cell secretion monitoring. In this work, we present the first cellulose microfluidic paper-based analytical device for the single-step detection of cell secreted Vascular Endothelial Growth Factor through a self-reporting Structure Switching Signaling Aptamer. A three-part Structure Switching Signaling Aptamer was designed with an aptameric sequence specific for VEGF, which provides a quantifiable fluorescent signal through the displacement of a quencher upon VEGF recognition. The VEGF biosensor was integrated in cellulose paper, enabling the homogenous distribution of the sensor in the paper substrate and the detection of as low as 0.34ng of VEGF in 30min through fluorescence intensity analysis. As a proof-of-concept, the biosensor was incorporated in a microfluidic paper-based analytical device format containing a VEGF detection zone and a control zone, which was applied for the detection of cell secreted VEGF in the supernatant of mesenchymal stem cells culture plates, demonstrating its potential use in cell biology research. Authors acknowledge funding support from Basque Government, under Grupos Consolidados with Grant No. IT1271-19, from “Ministerio de Ciencia y Educación de España” under grant PID2020-120313GB-I00 /AIE /10.13039/501100011033, from the University of the Basque Country and the Spanish Government under the program “Margarita Salas” funded by “Unión Europea-Next Generation EU” . LBD and FBL personally acknowledge funds from the DNASURF (H2020-MSCA-RISE-778001) project. The authors thank for technical and human support provided by PhD Maite Alvarez from DNA Bank Service (SGIker) of the University of the Basque Country (UPV/EHU) and European funding (ERDF and ESF). Authors also wish to thank the intellectual and technical assistance from the ICTS “NANBIOSIS”, more specifically by the Drug Formulation Unit (U10) of the CIBER in Bioengineering, Biomaterials & Nanomedicine (CIBER-BBN). Authors thank Dr. Alberto Gorrochategui from “Clínica Dermatológica Ercilla (Bilbao)” for providing the hair follicles from where hHF-MSCs were extracted. Finally, authors also wish to thank the technical assistance from the Nanopharmagene research group at the University of the Basque Country (UPV/EHU). FB-L and LBD acknowledge the “Red de Microfluídica Española” RED2018-102829-T.
- Published
- 2022
9. Paper-Based Cell Culture: Paving the Pathway for Liver Tissue Model Development on a Cellulose Paper Chip
- Author
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Mimi R. Borrelli, Tapas K. Maiti, Milad Ashrafizadeh, Tarun Agarwal, and Pooyan Makvandi
- Subjects
congenital, hereditary, and neonatal diseases and abnormalities ,education ,Biochemistry (medical) ,Biomedical Engineering ,General Chemistry ,Paper based ,Chip ,humanities ,Cell biology ,Biomaterials ,chemistry.chemical_compound ,chemistry ,Cell culture ,Liver tissue ,Model development ,Cellulose - Abstract
The present review provides a comprehensive outlook toward the possibilities of developing a functional in vitro liver tissue model on a paper platform. To this end, we first addressed the suitabil...
- Published
- 2022
10. In situ paper-based 3D cell culture for rapid screening of the anti-melanogenic activity.
- Author
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Pupinyo, Naricha, Chatatikun, Moragot, Chiabchalard, Anchalee, and Laiwattanapaisal, Wanida
- Subjects
CELL culture ,MELANOGENESIS ,BIOTECHNOLOGY research - Abstract
Recently, paper has gained traction in the biotechnology research field due to its ability to be a substrate for 3D cell culture. In this work, we demonstrate the application of paper-based 3D cell culture for rapid and easy screening of the effect of natural compounds on melanin production. Whatman No. 1 filter paper was used as the substrate for B16F10 melanoma cell culture. The use of paper is beneficial for supporting the 3D structure of cells, which makes the result more reliable due to the similarity to in vivo conditions. Furthermore, paper is beneficial for melanin observation due to melanin's black color, which is easily in situ visualized after it is cultured on white paper. Matrigel was used to encapsulate cells before being pipetted onto the paper to prevent the passing of cells through paper pores. The intensity of melanin can then be observed with the naked eye and analyzed by scanning the paper. The analysis process took only 20 minutes, which is faster than that of the conventional absorbance spectroscopy, owing to the elimination of centrifugation, melanin solubilization, and the absorbance measurement step. The color intensity on the paper showed a direct proportion with increased α-MSH concentrations, confirming that the color on the paper was melanin. The 3D structure of cells was confirmed by using a scanning electron microscope. To demonstrate the application of the paper-based scaffold, paper-based 3D cell culture was used for screening the effects of Kojic acid and Arbutin on melanin production, which showed increased anti-melanogenesis effects with increased concentrations of natural compounds. High cell viability was observed over 120 hours. In conclusion, the developed paper-based scaffold can be used for screening the effect of natural compounds on melanin production, as a rapid and simple method with low cost. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
11. Transparent Microcrystalline Cellulose/Polyvinyl Alcohol Paper as a New Platform for Three-Dimensional Cell Culture
- Author
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Fei Li, Ruihua Tang, Li Na Liu, Yaowei Yang, Min Li, Xue Yao, and Sufeng Zhang
- Subjects
Paper ,Biocompatibility ,Surface Properties ,Chemistry ,Water ,Biocompatible Materials ,Polyvinyl alcohol ,Analytical Chemistry ,Microcrystalline cellulose ,chemistry.chemical_compound ,Chemical engineering ,Cell culture ,Cell Line, Tumor ,Polyvinyl Alcohol ,Cell Adhesion ,Humans ,Cellulose ,Porosity ,Hydrophobic and Hydrophilic Interactions ,Cells, Cultured ,Cell Proliferation ,Mechanical Phenomena - Abstract
Multilayered and stacked cellulose paper has emerged as a promising platform for construction of three-dimensional (3D) cell culture because of its low cost, good biocompatibility, and high porosity. However, its poor light transmission makes it challenging to directly and clearly monitor cell behaviors (e.g., growth and proliferation) on the paper-based platform using an optical microscope. In this work, we developed a transparent microcrystalline cellulose/polyvinyl alcohol (MCC/PVA) paper with irregular pores through dissolution and regeneration of microcrystalline nanocellulose, addition of a porogen reagent (NaCl), and subsequently dipping in PVA solutions. The transparent MCC paper displays high porosity (up to 90%), adjustable pore size (between 23 and 46 μm), large thickness (from 315 to 436 μm), and high light transmission under water (95%). Through further modification of the transparent MCC paper with PVA, the obtained transparent MCC/PVA paper shows enhanced mechanical properties (dry and wet strengths), good hydrophilicity (with a contact angle of 70.8°), and improved biocompatibility (cell viability up to 90%). By stacking and destacking multiple layers of the transparent MCC/PVA paper, it has been used for both two-dimensional and three-dimensional cell culture platforms. The transparent MCC/PVA paper under water enables both direct observation of cell morphology by an optical microscope via naked eyes and fluorescence microscope after staining. We envision that the developed transparent MCC/PVA paper holds great potential for future applications in various bioanalytical and biomedical fields, such as drug screening, tissue engineering, and organ-on-chips.
- Published
- 2020
12. Low-cost electrochemical paper-based device for exosome detection.
- Author
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Kasetsirikul, Surasak, Tran, Kim Thinh, Clack, Kimberley, Soda, Narshone, Shiddiky, Muhammad J. A., and Nguyen, Nam-Trung
- Subjects
- *
EXOSOMES , *EXTRACELLULAR matrix , *DETECTION limit , *CELL culture , *OVARIAN cancer , *CANCER cells - Abstract
Exosomes are vesicles released by healthy and cancer cells into the extracellular matrix and bodily fluid. Cancer cell-derived exosomes have attracted much attention in early-stage detection and prognostication of treatment response. Thus, detecting exosomes is of great interest to biology and medicine. However, many conventional detection methods require high-cost equipment and centralized laboratory facilities, making diagnostics inaccessible in limited-resource settings. This study reports a proof-of-concept low-cost electrochemical paper-based analytical device to quantify both the total bulk and cancer cell-derived exosomes in cell culture media. The device employs a sandwich immune assay design, where exosomes are initially captured using the electrode-bound generic antibodies (i.e. CD9) and subsequently detected via ovarian cancer-specific CA125 antibodies. Our proposed device quantifies the total bulk exosome concentration with a detection limit of 9.3 × 107 exosomes per mL and ovarian cancer cell-derived exosomes with a detection limit of 7.1 × 108 exosomes per mL, with a relative standard deviation of <10% (n = 3). We suggest that this low-cost and simple electrochemical paper-based device could be an alternative tool for detecting disease-specific exosomes in biological samples with the potential to be further developed for point-of-care diagnosis. [ABSTRACT FROM AUTHOR]
- Published
- 2022
- Full Text
- View/download PDF
13. Multilayer three-dimensional filter paper constructs for the culture and analysis of aortic valvular interstitial cells.
- Author
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Sapp, Matthew C., Fares, Hannelle J., Estrada, Ana C., and Grande-Allen, K. Jane
- Subjects
FILTER paper ,INTERSTITIAL cells ,CELL culture ,COLLAGEN ,AORTIC valve diseases ,DISEASE progression - Abstract
Culturing aortic valvular interstitial cells in an environment that models the aortic valve is an essential step towards understanding the progression of calcific aortic valve disease. Here the adaption of a three-dimensional (3-D) stacked paper-based culture system is presented for analyzing valve cells in a thick collagen gel matrix. Filter paper layers, modeled after a 96-well plate design, were printed with a wax well-plate template and then seeded with valve cell and collagen mixtures that quickly gelled into 3-D cultures. Stacking these layers permitted extensive customization of culture thickness and cell density profiles to model the full thickness of native valve tissue. Aortic valvular interstitial cells seeded into the paper-based constructs consistently demonstrated high survival up to 14 days of culture with significant increases in cell number through the first 3 days of culture. After 4 days following seeding, valve cells in single layer cultures showed reduced smooth muscle α-actin expression with a stabilized cell density, suggesting a transition from an activated phenotype to a more quiescent state. Valve cells in multilayer cultures demonstrated the ability to migrate from layer to layer and had the highest smooth muscle α-actin expression in areas with predicted low oxygen tensions. These results establish the filter-paper-based method as a viable culture system for analyzing valve cells in an in vitro 3-D model of the aortic valve. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
14. Paper-based Transwell assays: an inexpensive alternative to study cellular invasion
- Author
-
Adam Loeser, Nathan A. Whitman, Matthew R. Lockett, and Rachael M. Kenney
- Subjects
Paper ,02 engineering and technology ,01 natural sciences ,Biochemistry ,Article ,Analytical Chemistry ,Extracellular matrix ,Cell Movement ,Cell Line, Tumor ,Electrochemistry ,Animals ,Humans ,Environmental Chemistry ,Neoplasm Invasiveness ,Spectroscopy ,Chemistry ,010401 analytical chemistry ,Exogenous factor ,Disease progression ,Reproducibility of Results ,Cell movement ,Paper based ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Cell biology ,Cell culture ,Biological Assay ,Cattle ,0210 nano-technology - Abstract
Cellular movement is essential in the formation and maintenance of healthy tissues as well as in disease progression such as tumor metastasis. In this work, we describe a paper-based Transwell assay capable of quantifying cellular invasion through an extracellular matrix. The paper-based Transwell assays generate similar datasets, with equivalent reproducibility, to commercially available Transwell assays. With different culture configurations, we quantify invasion: upon addition of an exogenous factor or in the presence of medium obtained from other cell types, in an indirect or direct co-culture format whose medium composition is dynamically changing, and in a single-zone or parallel (96-zone) format.
- Published
- 2019
15. In situ paper-based 3D cell culture for rapid screening of the anti-melanogenic activity
- Author
-
Naricha Pupinyo, Anchalee Chiabchalard, Wanida Laiwattanapaisal, and Moragot Chatatikun
- Subjects
Paper ,Cell Survival ,Cell Culture Techniques ,Drug Evaluation, Preclinical ,02 engineering and technology ,01 natural sciences ,Biochemistry ,Analytical Chemistry ,Melanin ,Absorbance ,Mice ,chemistry.chemical_compound ,3D cell culture ,Limit of Detection ,Cell Line, Tumor ,Electrochemistry ,Animals ,Environmental Chemistry ,Spectroscopy ,Melanins ,Matrigel ,Filter paper ,010401 analytical chemistry ,Arbutin ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,chemistry ,Pyrones ,alpha-MSH ,Cell culture ,Biophysics ,0210 nano-technology ,Kojic acid - Abstract
Recently, paper has gained traction in the biotechnology research field due to its ability to be a substrate for 3D cell culture. In this work, we demonstrate the application of paper-based 3D cell culture for rapid and easy screening of the effect of natural compounds on melanin production. Whatman No. 1 filter paper was used as the substrate for B16F10 melanoma cell culture. The use of paper is beneficial for supporting the 3D structure of cells, which makes the result more reliable due to the similarity to in vivo conditions. Furthermore, paper is beneficial for melanin observation due to melanin's black color, which is easily in situ visualized after it is cultured on white paper. Matrigel was used to encapsulate cells before being pipetted onto the paper to prevent the passing of cells through paper pores. The intensity of melanin can then be observed with the naked eye and analyzed by scanning the paper. The analysis process took only 20 minutes, which is faster than that of the conventional absorbance spectroscopy, owing to the elimination of centrifugation, melanin solubilization, and the absorbance measurement step. The color intensity on the paper showed a direct proportion with increased α-MSH concentrations, confirming that the color on the paper was melanin. The 3D structure of cells was confirmed by using a scanning electron microscope. To demonstrate the application of the paper-based scaffold, paper-based 3D cell culture was used for screening the effects of Kojic acid and Arbutin on melanin production, which showed increased anti-melanogenesis effects with increased concentrations of natural compounds. High cell viability was observed over 120 hours. In conclusion, the developed paper-based scaffold can be used for screening the effect of natural compounds on melanin production, as a rapid and simple method with low cost.
- Published
- 2019
16. Paper-based plasmonic substrates as surface-enhanced Raman scattering spectroscopy platforms for cell culture applications
- Author
-
L. Guerrini, Yanan Kang, K. Juarez-Moreno, José M. Romo-Herrera, R.A. Alvarez-Puebla, Wolfgang J. Parak, N. Feliu, and Publica
- Subjects
Medicine (General) ,Materials science ,Biocompatibility ,QH301-705.5 ,Biomedical Engineering ,Nanoparticle ,Bioengineering ,Nanotechnology ,Review Article ,Nanomaterials ,Biomaterials ,R5-920 ,Biology (General) ,Plasmonic nanoparticles ,Molecular Biology ,Plasmon ,SERS ,Paper-based substrates ,Cell Biology ,Plasmonic papers ,Colloidal gold ,Cell culture ,Biosensor ,Biotechnology - Abstract
The engineering of advanced materials capable of mimicking the cellular micro-environment while providing cells with physicochemical cues is central for cell culture applications. In this regard, paper meets key requirements in terms of biocompatibility, hydrophilicity, porosity, mechanical strength, ease of physicochemical modifications, cost, and ease of large-scale production, to be used as a scaffold material for biomedical applications. Most notably, paper has demonstrated the potential to become an attractive alternative to conventional biomaterials for creating two-dimensional (2D) and three-dimensional (3D) biomimetic cell culture models that mimic the features of in vivo tissue environments for improving our understanding of cell behavior (e.g. growth, cell migration, proliferation, differentiation and tumor metastasis) in their natural state. On the other hand, integration of plasmonic nanomaterials (e.g. gold nanoparticles) within the fibrous structure of paper opens the possibility to generate multifunctional scaffolds equipped with biosensing tools for monitoring different cell cues through physicochemical signals. Among different plasmonic based detection techniques, surface-enhanced Raman scattering (SERS) spectroscopy emerged as a highly specific and sensitive optical tool for its extraordinary sensitivity and the ability for multidimensional and accurate molecular identification. Thus, paper-based plasmonic substrates in combination with SERS optical detection represent a powerful future platform for monitoring cell cues during cell culture processes. To this end, in this review, we will describe the different methods for fabricating hybrid paper-plasmonic nanoparticle substrates and their use in combination with SERS spectroscopy for biosensing and, more specifically, in cell culture applications., Graphical abstract Image 1
- Published
- 2021
17. A Novel Wick-Like Paper-Based Microfluidic Device for 3D Cell Culture and Anti-Cancer Drugs Screening
- Author
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Shu‐xia Fu, Bang-Ce Ye, and Peng Zuo
- Subjects
Paper ,Materials science ,Microfluidics ,Cell Culture Techniques ,Nanotechnology ,Antineoplastic Agents ,General Medicine ,Paper based ,Substrate (printing) ,Microfluidic Analytical Techniques ,Applied Microbiology and Biotechnology ,3D cell culture ,Cell culture ,Lab-On-A-Chip Devices ,Anti cancer drugs ,Miniaturization ,Drug production ,Molecular Medicine ,Colorimetry - Abstract
Paper is increasingly recognized as a portable substrate for cell culture, due to its low-cost, flexible, and special porous property, which provides a native cellular 3D microenvironment. Therefore, paper-based microfluidics has been developed for cell culture and biomedical analysis. However, the inability of continuous medium supply limits the wide application of paper devices for cell culture. Herein, a paper-based microfluidic device is developed with novel folded paper strips as wick-like structure, which is used for medium self-driven perfusion. The paper with patterns of hydrophilic channel, culture areas, and hydrophobic barrier could be easily fabricated through wax-printing. After printing, the hydrophilic paper strip at the periphery of the lower layer is then folded at 90° and extended into the medium container for continuous automatic supply of medium to the cell culture area. Tumor cells cultured in the paper device are tested for anti-cancer drug screening. Visualized cell viability and chemical sensitivity testing can be achieved by colorimetry combined with simple smartphone imaging, effectively reducing precision instrument dependence. The wick paper-based microfluidic device for cell culture endows the method the advantages of lower cost, ease-of-operation, miniaturization, and shows a great potential for large-scale cell culture, antibody drug production, and efficient screening.
- Published
- 2020
18. A pH-Sensing Optode for Mapping Spatiotemporal Gradients in 3D Paper-Based Cell Cultures
- Author
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Brenden P. Kromhout, Nathan A. Whitman, Matthew R. Lockett, Rachael M. Kenney, and Matthew W. Boyce
- Subjects
Paper ,Polyurethanes ,Cell Culture Techniques ,Oligosaccharides ,Chitin ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Article ,Analytical Chemistry ,Cell Line, Tumor ,Extracellular ,Humans ,Fluorescent Dyes ,Anthracenes ,Chitosan ,Tumor microenvironment ,Chemistry ,Aggressive cancer ,Hydrogels ,Paper based ,Hydrogen-Ion Concentration ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Microscopy, Fluorescence ,Cell culture ,Biophysics ,Ph sensing ,Fluorescein ,Optode ,0210 nano-technology - Abstract
Paper-based cultures are an emerging platform for preparing 3D tissue-like structures. Chemical gradients can be imposed upon these cultures, generating microenvironments similar to those found in poorly vascularized tumors. There is increasing evidence that the tumor microenvironment is responsible for promoting drug resistance and increased invasiveness. Acidosis, or the acidification of the extracellular space, is particularly important in promoting these aggressive cancer phenotypes. To better understand how cells respond to acidosis there is a need for 3D culture platforms that not only model relevant disease states but also contain sensors capable of quantifying small molecules in the extracellular environment. In this work, we describe pH-sensing optodes that are capable of generating high spatial and temporal resolution maps of pH gradients in paper-based cultures. This sensor was fabricated by suspending microparticles containing pH-sensitive (fluorescein) and pH-insensitive (diphenylanthracene) dyes in a polyurethane hydrogel, which was then coated onto a transparent film. The pH-sensing films have a fast response time, are reversible, stable in long-term culture environments, have minimal photobleaching, and are not cytotoxic. These films have a pKa of 7.61 ± 0.04 and are sensitive in the pH range corresponding to normal and tumorigenic tissues. With these optodes, we measured the spatiotemporal evolution of pH gradients in paper-based tumor models.
- Published
- 2018
19. Aggregate Removal Nanofiltration of Human Serum Albumin Solution Using Nanocellulose-Based Filter Paper
- Author
-
Lulu Wu, Athanasios Mantas, Simon Gustafsson, Levon Manukyan, and Albert Mihranyan
- Subjects
virus removal filtration ,Cladophora cellulose ,plasma-derived biologics ,cell culture ,cell therapies ,Biology (General) ,QH301-705.5 - Abstract
This study is dedicated to the rapid removal of protein aggregates and viruses from plasma-derived human serum albumin (HSA) product to reduce the risk of viral contamination and increase biosafety. A two-step filtration approach was implemented to first remove HSA aggregates and then achieve high model virus clearance using a nanocellulose-based filter paper of different thicknesses, i.e., 11 μm (prefilter) and 22 μm (virus filter) at pH 7.4 and room temperature. The pore size distribution of these filters was characterized by nitrogen gas sorption analysis. Dynamic light scattering (DLS) and size-exclusion high performance liquid chromatography (SE-HPLC) were performed to analyze the presence of HSA aggregates in process intermediates. The virus filter showed high clearance of a small-size model virus, i.e., log10 reduction value (LRV) > 5, when operated at 3 and 5 bar, but a distinct decrease in LRV was detected at 1 bar, i.e., LRV 2.65–3.75. The throughput of HSA was also dependent on applied transmembrane pressure as was seen by Vmax values of 110 ± 2.5 L m−2 and 63.6 ± 5.8 L m−2 at 3 bar and 5 bar, respectively. Protein loss was low, i.e., recovery > 90%. A distribution of pore sizes between 40 nm and 60 nm, which was present in the prefilter and absent in the virus filter, played a crucial part in removing the HSA aggregates and minimizing the risk of virus filter fouling. The presented results enable the application of virus removal nanofiltration of HSA in bioprocessing as an alternative to virus inactivation methods based, e.g., on heat treatment.
- Published
- 2020
- Full Text
- View/download PDF
20. A Hybrid Nanofiber/Paper Cell Culture Platform for Building a 3D Blood–Brain Barrier Model.
- Author
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Huang, Kaixiang, Castiaux, Andre D., Podicheti, Ram, Rusch, Douglas B., Martin, R. Scott, and Baker, Lane A.
- Subjects
- *
CELL culture , *BLOOD-brain barrier , *PLURIPOTENT stem cells , *TIGHT junctions , *CENTRAL nervous system , *BASAL lamina , *PHENOTYPES - Abstract
The blood–brain barrier (BBB) protects the central nervous system from toxins and pathogens in the blood by regulating permeation of molecules through the barrier interface. In vitro BBB models described to date reproduce some aspects of BBB functionality, but also suffer from incomplete phenotypic expression of brain endothelial traits, difficulty in reproducibility and fabrication, or overall cost. To address these limitations, a 3D BBB model based on a hybrid paper/nanofiber scaffold is described. The cell culture platform utilizes lens paper as a framework to accommodate 3D culture of astrocytes. An electrospun nanofiber layer is coated onto one face of the paper to mimic the basement membrane and support growth of an organized 2D layer of endothelial cells (ECs). Human induced pluripotent stem cell‐derived ECs and astrocytes are co‐cultured to develop a human BBB model. Morphological and spatial organization of model are validated with confocal microscopy. Measurements of transendothelial resistance and permeability demonstrate the BBB model develops a high‐quality barrier and responds to hyperosmolar treatments. RNA‐sequencing shows introduction of astrocytes both regulates EC tight junction proteins and improves endothelial phenotypes related to vasculogenesis. This model shows promise as a model platform for future in vitro studies of the BBB. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
21. Paper-Based 3D Scaffold for Multiplexed Single Cell Secretomic Analysis
- Author
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Chunyue Miao, Ruihan Bai, Shiqiang Yan, Bingcheng Lin, Linmei Li, Tingjiao Liu, Yibing Ji, Yao Lu, Yong Luo, Ruijun Li, and Meimei Liu
- Subjects
Paper ,0301 basic medicine ,Scaffold ,Cell ,02 engineering and technology ,Matrix (biology) ,Analytical Chemistry ,03 medical and health sciences ,Tumor Cells, Cultured ,medicine ,Extracellular ,Humans ,Secretion ,U87 ,Chemistry ,021001 nanoscience & nanotechnology ,030104 developmental biology ,medicine.anatomical_structure ,Membrane ,Cellular Microenvironment ,Cell culture ,Printing, Three-Dimensional ,Biophysics ,Polyvinyls ,Single-Cell Analysis ,0210 nano-technology - Abstract
Despite rapid progresses in single-cell analysis technologies, efforts to control the three-dimensional microenvironment for single cell measurements have been lacking. Here, we report a simple method to incorporate three-dimensional scaffolds, including polyvinylidene fluoride (PVDF) membranes and PVDF membrane replicated analog polydimethylsiloxane, into multiplexed single cell secretomic analysis platforms (including a microwell array and a single cell barcode microchip) to mimic the extracellular physical matrix and mechanical support for single cells. Applying this platform to brain tumor cell line U87 to investigate single cell protein secretion behavior on different substrates, we revealed that single cell protein secretions were regulated differently in three-dimensional (3D) microenvironments. This finding was further verified with intracellular cytokine staining, highlighting the significance of 3D single cell microenvironments. This new single cell biomimetic platform can be easily adaptable to other three-dimensional cell culture scaffolds or other single cell assays and may become a broadly applicable three-dimensional single cell analysis system to study the effect of microenvironment conditions on cellular functional heterogeneity in vitro.
- Published
- 2018
22. Exosome aggregation mediated stop-flow paper-based portable device for rapid exosome quantification
- Author
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Leena Suntornsuk, Nantana Nuchtavorn, Yong Zeng, and Boonta Chutvirasakul
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Paper ,Clinical Biochemistry ,Cytological Techniques ,Nanoparticle tracking analysis ,02 engineering and technology ,Exosomes ,01 natural sciences ,Biochemistry ,Exosome ,Analytical Chemistry ,Tetraspanin 28 ,Limit of Detection ,Cell Line, Tumor ,Humans ,Chemistry ,Stop flow ,Vesicle ,010401 analytical chemistry ,Reproducibility of Results ,Paper based ,Microfluidic Analytical Techniques ,021001 nanoscience & nanotechnology ,Microvesicles ,Polyacetylene Polymer ,0104 chemical sciences ,Cell culture ,Biophysics ,Linear Models ,Naked eye ,0210 nano-technology - Abstract
Exosome quantification is important for estimation of informative messengers (e.g., proteins, lipids, RNA, etc.) involving physiological and pathological effects. This work aimed to develop a simple and rapid distance-based paper portable device using exosome-capture vesicles (polydiacetylene conjugated with antiCD81) for exosome quantification in cell cultures. This novel concept relied on distinct aggregation of exosomes and exosome-capture vesicles leading to different solvent migration. Distances of the migration were used as signal readouts, which could be detected by naked eye. PDA-antiCD81 as exosome-capture vesicles were optimized (e.g., size, reaction ratio, and concentration) and the paper designs were investigated (e.g., diameter of sample reservoir and lamination layer) to enhance the solvent stop-flow effects. Finally, exosome screening on three cell culture samples (COLO1, MDA-MB-231, and HuR-KO1 subclone) was demonstrated. The method could linearly measure exosome concentrations in correlation with solvent migration distances in the range of 106 -1010 particles/mL (R2 > 0.98) from the cell culture samples. The exosome concentration measurements by the developed device were independently assessed by nanoparticle tracking analysis. Results demonstrated no statistically significant difference (p > 0.05) by t-test. This low-cost and rapid device allows a portable platform for exosome quantification without the requirement of expensive equipment and expertise of operation. The developed device could potentially be useful for quantification of other biomarker-related extracellular vesicles.
- Published
- 2019
23. Paper-based cell culture platform and its emerging biomedical applications
- Author
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Xiaohui Zhang, Zedong Li, Xin Zhao, Kar Wey Yong, Feng Xu, Yuhui Li, Belinda Pingguan-Murphy, Meng Shi, Kelvin Ng, Bin Gao, and Hui Yang
- Subjects
business.industry ,Computer science ,Mechanical Engineering ,Nanotechnology ,02 engineering and technology ,Paper based ,010402 general chemistry ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,01 natural sciences ,Intervention studies ,Cell Microenvironment ,0104 chemical sciences ,3D cell culture ,Materials Science(all) ,Cell culture ,Mechanics of Materials ,General Materials Science ,0210 nano-technology ,business ,Biomedicine - Abstract
Paper has recently attracted increasing attention as a substrate for various biomedical applications. By modifying its physical and chemical properties, paper can be used as an alternative to conventional cell culture substrates. Because it can be stacked into a three-dimensional (3D) structure, which can better mimic the in vivo cell microenvironment. Paper has shown great potential as a 3D cell culture platform for developing normal and diseased models. This platform gives precise control over extracellular matrix (ECM) composition as well as cell distribution and precise analysis of the interactions between cells. Paper-based platforms have been applied for pathophysiological studies and therapeutic intervention studies. In this paper, we first discuss the modifications of the physical and chemical properties of paper to develop various 2D and 3D cell culture platforms. We then review the applications of paper-based cell culture platforms for the construction of in vitro disease models, drug screening, and cell cryopreservation applications. Because of its advantages such as biocompatibility, eco-friendliness, cost efficiency, and ease of large-scale production, we believe that paper-based cell culture platforms would play an important role in the fields of biomedicine.
- Published
- 2017
- Full Text
- View/download PDF
24. Redefining Chinese calligraphy rice paper: an economical and cytocompatible substrate for cell biological assays
- Author
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Ying Shuai Liu, Yuejun Kang, Ying Zhou, Ling Yu, Chang Ming Li, and Jing Jing Fu
- Subjects
0301 basic medicine ,Filter paper ,Cell growth ,Chemistry ,General Chemical Engineering ,Petri dish ,Cell ,food and beverages ,Substrate (chemistry) ,Nanotechnology ,02 engineering and technology ,General Chemistry ,021001 nanoscience & nanotechnology ,law.invention ,03 medical and health sciences ,Cellulose fiber ,030104 developmental biology ,medicine.anatomical_structure ,DU145 ,Cell culture ,law ,medicine ,0210 nano-technology - Abstract
Paper is a permeable porous material composed of a solid network of fibers. It is cheap, abundant, disposable and recyclable and has self-powered fluid wicking properties that are useful in building analytical devices. Paper-based cell assays are still in their infancy compared with enzyme- and protein-based analyses. For the first time, we show the potential of rice paper (an organic paper specifically used in Chinese calligraphy) for building cell analysis platforms. Rice paper's solution wicking and surface characterizations prove that it has a similar chemical configuration as that of a standard Whatman filter paper. Moreover, lactate dehydrogenase (LDH) release assay and WST-1 cell growth assay show that rice paper has better cell-compatibility features and improved cell distribution. The cell anchors and spreads along the cellulose fiber of the rice paper, whereas the porous rice paper matrix provides a sufficient surface area for cell growth. Cell-based immunohistochemistry was conducted to measure the expression of O-linked N-acetylglucosamine (O-GlcNAc) protein on prostate cancer cell DU145. An enhanced colorimetric signal was observed from cells grown on rice paper-based cell culture platform than those grown on 2D culture dish. The feasibility of fabricating rice paper with both direct crafting and wax printing—as well as on-paper cell immunoassays for on-demand applications—confirms the potential of rice paper as a new substrate for building paper devices for cell biology studies.
- Published
- 2017
25. Simple Paper-based Liver Cell Model for Drug Screening
- Author
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Christina Loera, Kattika Kaarj, Patarajarin Akarapipad, Jennifer Ngo, Soohee Cho, and Jeong Yeol Yoon
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Drug ,Chemistry ,Liver cell ,media_common.quotation_subject ,010401 analytical chemistry ,Biomedical Engineering ,Antifungal drug ,Bioengineering ,02 engineering and technology ,Pharmacology ,021001 nanoscience & nanotechnology ,Cell morphology ,01 natural sciences ,Organ-on-a-chip ,0104 chemical sciences ,Drug development ,Cell culture ,Toxicity ,Electrical and Electronic Engineering ,0210 nano-technology ,Biotechnology ,media_common - Abstract
Investigation of the potential adverse effects of chemicals and drugs is essential during the drug development process. In vitro cell model systems have been developed over the past years towards such toxicity investigation. 96-well plate is the common platform for screening drug toxicity due to its simplicity. However, this platform only offers 2D cell culture environment and lacks the flow of solutions, which fails to provide the suitable environment for the cells to adequately metabolize the drugs, for the media to replenish, and for the metabolites and wastes to be removed. Microfluidic chips populated with human or animal cells, known as organ-on-a-chip (OOC), can reconcile many issues of in vitro cell models, such as the lack of extracellular matrix and flow as well as the species difference. However, OOC can be complicated to fabricate and operate. To bridge this gap, we utilized paper as a primary substrate for OOC, considering its fibrous structure that can mimic natural extracellular matrix, as well as a syringe pump and filter that are commonly available in most laboratories. Paper microfluidic model was designed and fabricated by wax printing on nitrocellulose paper, seeded and proliferated with liver cells (primary rat hepatocytes and HepG2 cells), and two paper substrates were stacked together to complete the paper model. To this paper-based liver cell model, the following drugs were added: Phenacetin (pain reliever and fever reducer), Bupropion (antidepressant), Dextromethorphan (antidepressant), and phosphate-buffered saline (PBS) as a control, all under a physiologically relevant flow rate. The combination of these drugs with Fluconazole (antifungal drug) was also investigated. Cell count, cell morphology, protein production, and urea secretion after drug treatment confirmed that the model successfully predicted toxicity within 40 minutes. This simple, paper-based liver cell model provided enhanced and faster cell response to drug toxicity and showed comparable or better behavior than the cells cultured in conventional 2D in vitro models.
- Published
- 2020
26. Analysis of the Growth Process of Neural Cells in Culture Environment Using Image Processing Techniques
- Author
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Mirsafianf, Atefeh S., Isfahani, Shirin N., Kasaei, Shohreh, Mobasheri, Hamid, Sarbazi-Azad, Hamid, editor, Parhami, Behrooz, editor, Miremadi, Seyed-Ghassem, editor, and Hessabi, Shaahin, editor
- Published
- 2009
- Full Text
- View/download PDF
27. Evaluation of Hybrid Vesicles in an Intestinal Cell Model Based on Structured Paper Chips
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Noga Gal, Isabella Nymann Westensee, Paula De Dios Andres, Brigitte Städler, Miguel A. Ramos-Docampo, and Edit Brodszkij
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Cell type ,Polymers and Plastics ,Chemistry ,Vesicle ,Cell ,Nanoparticle ,Bioengineering ,Methacrylate ,Mucus ,Biomaterials ,Intestines ,medicine.anatomical_structure ,Cell culture ,Amphiphile ,Materials Chemistry ,medicine ,Biophysics ,Humans ,Caco-2 Cells ,Intestinal Mucosa ,HT29 Cells - Abstract
Cell culture-based intestinal models are important to evaluate nanoformulations intended for oral drug delivery. We report the use of a floating structured paper chip as a scaffold for Caco-2 cells and HT29-MTX-E12 cells that are two established cell types used in intestinal cell models. The formation of cell monolayers for both mono- and cocultures in the paper chip are confirmed and the level of formed cell-cell junctions is evaluated. Further, cocultures show first mucus formation between 6-10 days with the mucus becoming more pronounced after 19 days. Hybrid vesicles (HVs) made from phospholipids and the amphiphilic block copolymer poly(cholesteryl methacrylate)-block-poly(2-carboxyethyl acrylate) in different ratios are used as a representative soft nanoparticle to assess their mucopenetration ability in paper chip-based cell cultures. The HV assembly is characterized, and it is illustrated that these HVs cross the mucus layer and are found intracellularly within 3 h when the cells are grown in the paper chips. Taken together, the moist three-dimensional cellulose environment of structured paper chips offers an interesting cell culture-based intestinal model that can be further integrated with fluidic systems or online read-out opportunities.
- Published
- 2021
28. Paper-based microreactor array for rapid screening of cell signaling cascades
- Author
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Kin Fong Lei, Ngan-Ming Tsang, and Chia-Hao Huang
- Subjects
Paper ,0301 basic medicine ,Cell signaling ,Time Factors ,Biomedical Engineering ,Bioengineering ,Biology ,01 natural sciences ,Biochemistry ,03 medical and health sciences ,Bioreactors ,Cell Line, Tumor ,Neoplasms ,Humans ,Transcription factor ,Cell Proliferation ,Immunoassay ,Kinase ,Liver Neoplasms ,010401 analytical chemistry ,Hydrogels ,Equipment Design ,General Chemistry ,Paper based ,Microarray Analysis ,0104 chemical sciences ,Cell biology ,030104 developmental biology ,Cell culture ,Immunology ,Computer-Aided Design ,Microreactor ,Signal transduction ,Cell signaling pathways ,Signal Transduction - Abstract
Investigation of cell signaling pathways is important for the study of pathogenesis of cancer. However, the related operations used in these studies are time consuming and labor intensive. Thus, the development of effective therapeutic strategies may be hampered. In this work, gel-free cell culture and subsequent immunoassay has been successfully integrated and conducted in a paper-based microreactor array. Study of the activation level of different kinases of cells stimulated by different conditions, i.e., IL-6 stimulation, starvation, and hypoxia, was demonstrated. Moreover, rapid screening of cell signaling cascades after the stimulations of HGF, doxorubicin, and UVB irradiation was respectively conducted to simultaneously screen 40 kinases and transcription factors. Activation of multi-signaling pathways could be identified and the correlation between signaling pathways was discussed to provide further information to investigate the entire signaling network. The present technique integrates most of the tedious operations using a single paper substrate, reduces sample and reagent consumption, and shortens the time required by the entire process. Therefore, it provides a first-tier rapid screening tool for the study of complicated signaling cascades. It is expected that the technique can be developed for routine protocol in conventional biological research laboratories.
- Published
- 2016
29. Real-time imaging of cancer cell chemotaxis in paper-based scaffolds
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Rachael M. Kenney, Matthew R. Lockett, Andrew S. Truong, Matthew W. Boyce, and C. Robert Bagnell
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Paper ,0301 basic medicine ,Time Factors ,Nanotechnology ,02 engineering and technology ,Biochemistry ,Analytical Chemistry ,03 medical and health sciences ,Gentamicin protection assay ,In vivo ,Cell Line, Tumor ,Electrochemistry ,Fluorescence microscope ,Humans ,Environmental Chemistry ,Neoplasm Invasiveness ,Spectroscopy ,Migration Assay ,Chemistry ,Chemotaxis ,Cell migration ,Hypoxia-Inducible Factor 1, alpha Subunit ,021001 nanoscience & nanotechnology ,Molecular Imaging ,Oxygen ,Protein Transport ,030104 developmental biology ,Cell culture ,Cancer cell ,Biophysics ,0210 nano-technology - Abstract
Cellular migration is the movement of cells, cultured as a monolayer; cellular invasion is similar to migration, but requires the cells to move through a three-dimensional material such as basement membrane extract or a synthetic hydrogel. Migration assays, such as the transwell assay, are widely used to study cellular movement because they are amenable to high-throughput screens with minimal experimental setup. These assays offer limited information about cellular responses to gradients in vivo because they oversimplify the threedimensional (3D) environment of a tissue. There are a number of invasion assays that support 3D cultures, some of which provide experimental control over the spatial and temporal gradients imparted on the culture. These assays, in their current form, are difficult to setup and maintain, and often require specialized laboratory equipment or engineering expertise. Here we describe a paper-based invasion assay in which cellular movement can be monitored in real-time with fluorescence microscopy. These assays are easily prepared and utilize materials commonly found in any laboratory: a single sheet of paper. These sheets are wax patterned to contain channels in which cells suspended in a hydrogel are seeded and cultured. Cell-containing sheets of paper are placed in a custom-built holder that allows gradients to form along the length of the channels. In this work, we compare the invasion of cells cultured in the presence and absence of an oxygen gradient. Our result support previous findings that oxygen is a chemoattractant, and selectively directs cellular movement in a 3D culture environment.
- Published
- 2016
30. A Hybrid Nanofiber/Paper Cell Culture Platform for Building a 3D Blood-brain Barrier Model
- Author
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Ram Podicheti, R. Scott Martin, Andre D Castiaux, Lane A. Baker, Douglas B. Rusch, and Kaixiang Huang
- Subjects
Basement membrane ,Tight junction ,Chemistry ,General Chemistry ,Blood–brain barrier ,Article ,law.invention ,Cell biology ,3D cell culture ,medicine.anatomical_structure ,Vasculogenesis ,Confocal microscopy ,law ,Cell culture ,medicine ,General Materials Science ,Induced pluripotent stem cell - Abstract
The blood brain barrier (BBB) protects the central nervous system from toxins and pathogens in the blood by regulating permeation of molecules through the barrier interface. In vitro BBB models described to date reproduce some aspects of BBB functionality, but also suffer from incomplete phenotypic expression of brain endothelial traits, difficulty in reproducibility and fabrication, or overall cost. To address these limitations, we describe a three-dimensional (3D) BBB model based on a hybrid paper/nanofiber scaffold. The cell culture platform utilizes lens paper as a framework to accommodate 3D culture of astrocytes. An electrospun nanofiber layer is coated onto one face of the paper to mimic the basement membrane and support growth of an organized two-dimensional layer of endothelial cells (ECs). Human induced pluripotent stem cell-derived ECs and astrocytes are co-cultured to develop a human BBB model. Morphological and spatial organization of model are validated with confocal microscopy. Measurements of transendothelial resistance and permeability demonstrate the BBB model develops a high-quality barrier and responds to hyperosmolar treatments. RNA-sequencing shows introduction of astrocytes both regulates EC tight junction proteins and improves endothelial phenotypes related to vasculogenesis. This model shows promise as a model platform for future in vitro studies of the BBB.
- Published
- 2021
31. Quantifying oxygen in paper-based cell cultures with luminescent thin film sensors
- Author
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Andrew S. Truong, Matthew W. Boyce, Rachael M. Kenney, and Matthew R. Lockett
- Subjects
Paper ,Fluorescence-lifetime imaging microscopy ,Luminescence ,010401 analytical chemistry ,Cell Culture Techniques ,Spheroid ,chemistry.chemical_element ,Nanotechnology ,02 engineering and technology ,021001 nanoscience & nanotechnology ,01 natural sciences ,Biochemistry ,Oxygen ,0104 chemical sciences ,Analytical Chemistry ,Oxygen tension ,chemistry.chemical_compound ,chemistry ,Cell culture ,Biophysics ,Polystyrene ,Thin film ,0210 nano-technology ,Oxygen sensor - Abstract
Paper-based scaffolds are an attractive material for generating 3D tissue-like cultures because paper is readily available and does not require specialized equipment to pattern, cut, or use. By controlling the exchange of fresh culture medium with the paper-based scaffolds, we can engineer diffusion-dominated environments similar to those found in spheroids or solid tumors. Oxygen tension directly regulates cellular phenotype and invasiveness through hypoxia-inducible transcription factors and also has chemotactic properties. To date, gradients of oxygen generated in the paper-based cultures have relied on cellular response-based readouts. In this work, we prepared a luminescent thin film capable of quantifying oxygen tensions in apposed cell-containing paper-based scaffolds. The oxygen sensors, which are polystyrene films containing a Pd(II) tetrakis(pentafluorophenyl)porphyrin dye, are photostable, stable in culture conditions, and not cytotoxic. They have a linear response for oxygen tensions ranging from 0 to 160 mmHg O2, and a Stern-Volmer constant (K sv) of 0.239 ± 0.003 mmHg O2 −1. We used these oxygen-sensing films to measure the spatial and temporal changes in oxygen tension for paper-based cultures containing a breast cancer line that was engineered to constitutively express a fluorescent protein. By acquiring images of the oxygen-sensing film and the fluorescently labeled cells, we were able to approximate the oxygen consumption rates of the cells in our cultures.
- Published
- 2015
32. An animal cell culture monitoring system using a smartphone-mountable paper-based analytical device
- Author
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Seong Hyun Im, Ka Ram Kim, Jung Woo Hong, Hyun C. Yoon, Yoo Min Park, and Jae Ho Yoon
- Subjects
Capillary action ,Microfluidics ,Analytical chemistry ,02 engineering and technology ,01 natural sciences ,Signal ,Horseradish peroxidase ,Materials Chemistry ,Electrical and Electronic Engineering ,Instrumentation ,Detection limit ,Chromatography ,biology ,Chemistry ,010401 analytical chemistry ,Metals and Alloys ,Monitoring system ,Paper based ,021001 nanoscience & nanotechnology ,Condensed Matter Physics ,0104 chemical sciences ,Surfaces, Coatings and Films ,Electronic, Optical and Magnetic Materials ,Cell culture ,biology.protein ,0210 nano-technology - Abstract
We developed a simple and low-cost cell culture monitoring system utilizing a paper-based analytical device (PAD) and a smartphone. The PAD simultaneously analyses glucose and lactate concentrations in the cell culture medium. Focusing on the fact that animal cells consume glucose and produce lactate under anaerobic conditions, oxidase- and horseradish peroxidase (HRP) enzyme-mediated colorimetric assays were integrated into the PAD. The PAD was designed to have three laminated layers. By using a double-sided adhesive tape as the middle layer and wax coating, a bifurcated fluidic channel was prepared to manipulate sample flow. At the inlet and the outlets of the channel, a sample drop zone and two detection zones for glucose and lactate, respectively, were positioned. When sample solution is loaded onto the drop zone, it flows to the detection zone through the hydrophilic fluidic channel via capillary force. Upon reaching the detection zone, the sample reacts with glucose and lactate oxidases (GOx and LOx) and HRP, immobilized on the detection zone along with colorless chromophores. By the Trinder’s reaction, the colorless chromophore is converted to a blue-colored product, generating concentration-dependent signal. With a gadget designed to aid the image acquisition, the PAD was positioned to the smartphone-embedded camera. Images of the detection zones were acquired using a mobile application and the color intensities were quantified as sensor signals. For the glucose assay using GOx/HRP format, we obtained the limit of detection (LOD ∼0.3 mM) and the limit of quantification (LOQ ∼0.9 mM) values in the dynamic detection range from 0.3 to 8.0 mM of glucose. For lactate assay using LOx/HRP, the LOD (0.02 mM) and the LOQ (0.06 mM) values were registered in the dynamic detection range from 0.02 to 0.50 mM of lactate. With the device, simultaneous analyses of glucose and lactate in cell culture media were conducted, exhibiting highly accurate and reproducible results. Based on the results, we propose that the optical sensing system developed is feasible for practical monitoring of animal cell culture.
- Published
- 2016
33. Title of presented paper: Isolation, breeding and characteristic of cells taken from the pulp of the tooth use in regeneration medicine.
- Author
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Jeżewski, Dawid, Nowak, Nicole, and Bubak, Joanna
- Subjects
REGENERATIVE medicine ,PAPER industry ,CELL transplantation ,ORGANOIDS ,EMBRYONIC stem cells - Abstract
Introduction and aim. Regenerative medicine includes various types of therapy, including transplantation of cells derived from embryonic stem cells, induced pluripotent stem cells, mobilization of endogenous stem cells, tissue engineering using natural scaffolds or organoids made from adult stem cells and pluripotent cells. The aim of the work will be to multiply stem cells located in the pulp, which can then be used in regenerative processes in the body. Material and methods. Isolation of cell line was conducted by collecting the material, and then via mechanical fragmentation of the material, cleaning the collected dental pulp. The next step was transferring the material to the test tubes with the medium NutriStem F Basal Medium with FBS in concentration 20% that is stored in the incubator in a humidified atmosphere of 95% air and 5% CO2 for 3 h at 37 °C. Results. In this research dental pulp stem cell line was established, by harvesting tissue samples from human volunteers and conducting isolation and cell culture. Conclusion. Established dental pupl stem cell line gives many possibilities in regenerative medicine, because it can be used in the regeneration of bone defects, corneal regeneration and neurodegenerative diseases. [ABSTRACT FROM AUTHOR]
- Published
- 2023
34. Engineering Cell-Compatible Paper Chips for Cell Culturing, Drug Screening, and Mass Spectrometric Sensing
- Author
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Ziyi He, Qiushui Chen, Wu Liu, Jin-Ming Lin, Xuexia Lin, Jing Wu, and Haifang Li
- Subjects
Paper ,Fluorescence-lifetime imaging microscopy ,Materials science ,Cell ,Cell Culture Techniques ,Drug Evaluation, Preclinical ,Biomedical Engineering ,Pharmaceutical Science ,Biocompatible Materials ,Nanotechnology ,Drug efficiency ,Mass spectrometry ,Mass Spectrometry ,Biomaterials ,Mice ,Adherent cell ,Cell Adhesion ,medicine ,Animals ,Cell adhesion ,Cell Engineering ,Polycarboxylate Cement ,Mass spectrometric ,medicine.anatomical_structure ,Cell culture ,NIH 3T3 Cells ,Biological Assay - Abstract
Paper-supported cell culture is an unprecedented development for advanced bioassays. This study reports a strategy for in vitro engineering of cell-compatible paper chips that allow for adherent cell culture, quantitative assessment of drug efficiency, and label-free sensing of intracellular molecules via paper spray mass spectrometry. The polycarbonate paper is employed as an excellent alternative bioscaffold for cell distribution, adhesion, and growth, as well as allowing for fluorescence imaging without light scattering. The cell-cultured paper chips are thus amenable to fabricate 3D tissue construction and cocultures by flexible deformation, stacks and assembly by layers of cells. As a result, the successful development of cell-compatible paper chips subsequently offers a uniquely flexible approach for in situ sensing of live cell components by paper spray mass spectrometry, allowing profiling the cellular lipids and quantitative measurement of drug metabolism with minimum sample pretreatment. Consequently, the developed paper chips for adherent cell culture are inexpensive for one-time use, compatible with high throughputs, and amenable to label-free and rapid analysis.
- Published
- 2015
35. A paper-based invasion assay: Assessing chemotaxis of cancer cells in gradients of oxygen
- Author
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Karl Patrick Gilbert, David Newsome, Howard Li, George M. Whitesides, Matthew R. Lockett, Kyaw Thu Minn, Hillier Shawn M, Karen A. Simon, Brenda K. Eustace, Diane M. Boucher, Amy B. Hall, and Bobak Mosadegh
- Subjects
Male ,Paper ,Cell Survival ,Cell Culture Techniques ,Biophysics ,Analytical chemistry ,Mice, Nude ,chemistry.chemical_element ,Bioengineering ,Oxygen ,Hydrogel, Polyethylene Glycol Dimethacrylate ,Permeability ,Biomaterials ,Mice ,Gentamicin protection assay ,Stack (abstract data type) ,Cell Line, Tumor ,Neoplasms ,Animals ,Humans ,Neoplasm Invasiveness ,Neoplasm Metastasis ,Chemotactic Factors ,Chemotaxis ,Models, Theoretical ,HEK293 Cells ,Phenotype ,chemistry ,Mechanics of Materials ,Permeability (electromagnetism) ,Cell culture ,Cancer cell ,Ceramics and Composites ,Biological Assay ,Layer (electronics) - Abstract
This work describes a 3D, paper-based assay that can isolate sub-populations of cells based on their invasiveness (i.e., distance migrated in a hydrogel) in a gradient of concentration of oxygen (O2). Layers of paper impregnated with a cell-compatible hydrogel are stacked and placed in a plastic holder to form the invasion assay. In most assays, the stack comprises a single layer of paper containing mammalian cells suspended in a hydrogel, sandwiched between multiple layers of paper containing only hydrogel. Cells in the stack consume and produce small molecules; these molecules diffuse throughout the stack to generate gradients in the stack, and between the stack and the bulk culture medium. Placing the cell-containing layer in different positions of the stack, or modifying the permeability of the holder to oxygen or proteins, alters the profile of the gradients within the stack. Physically separating the layers after culture isolates sub-populations of cells that migrated different distances, and enables their subsequent analysis or culture. Using this system, three independent cell lines derived from A549 cancer cells are shown to produce distinguishable migration behavior in a gradient of oxygen. This result is the first experimental demonstration that oxygen acts as a chemoattractant for cancer cells.
- Published
- 2015
36. Paper-based cell culture microfluidic system
- Author
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Kin Fong Lei, Xia Xiao, Fang Fang Tao, and I-Chi Lee
- Subjects
Computer science ,Cell growth ,Cell culture ,Untrained Personnel ,Microfluidics ,Biomedical Engineering ,Bioengineering ,Nanotechnology ,Paper based ,Electrical and Electronic Engineering ,Cell morphology ,Biotechnology - Abstract
In the past decades, glass/PDMS-based microfluidic systems have been rapidly developed to provide homogenous and stable microenvironment for culturing cells. Although these excellent demonstrations involve much simplified operations than traditional cell culture protocol, but they are still not readily accessible to untrained personnel and not appropriate to operate in conventional biological laboratories. In this work, cellulose filter papers were used for the substrates of the cell culture microfluidic system, which provides a convenient tool for cell-based assay. A paper was patterned with culture areas and channels by wax printing technique. Medium or tested substance can be passively perfused to the culture areas. Analyses of cyto-compatibility, cell proliferation, cell morphology, and cell chemosensitivity were performed to confirm the possibility of the paper-based system. The culture system could provide a platform for a wide range of cell-based assays with applications in drug screening and quantitative cell biology. This work demonstrated a paper-based cell culture microfluidic system and the system is inexpensive, disposable, and compatible to the existing culture facility.
- Published
- 2015
37. Paper-based Microreactor Integrating Cell Culture and Subsequent Immunoassay for the Investigation of Cellular Phosphorylation
- Author
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Kin Fong Lei and Chia-Hao Huang
- Subjects
Paper ,Materials science ,medicine.medical_treatment ,Cell Culture Techniques ,Biosensing Techniques ,Sensitivity and Specificity ,Cell Line, Tumor ,medicine ,Humans ,General Materials Science ,Immunoassay ,medicine.diagnostic_test ,Reproducibility of Results ,Equipment Design ,Neoplasms, Experimental ,Microfluidic Analytical Techniques ,Equipment Failure Analysis ,Systems Integration ,Blot ,Cytokine ,Biochemistry ,Cell culture ,Cancer cell ,Cytokines ,Phosphorylation ,Microreactor ,Signal transduction - Abstract
Investigation of cellular phosphorylation and signaling pathway has recently gained much attention for the study of pathogenesis of cancer. Related conventional bioanalytical operations for this study including cell culture and Western blotting are time-consuming and labor-intensive. In this work, a paper-based microreactor has been developed to integrate cell culture and subsequent immunoassay on a single paper. The paper-based microreactor was a filter paper with an array of circular zones for running multiple cell cultures and subsequent immunoassays. Cancer cells were directly seeded in the circular zones without hydrogel encapsulation and cultured for 1 day. Subsequently, protein expressions including structural, functional, and phosphorylated proteins of the cells could be detected by their specific antibodies, respectively. Study of the activation level of phosphorylated Stat3 of liver cancer cells stimulated by IL-6 cytokine was demonstrated by the paper-based microreactor. This technique can highly reduce tedious bioanalytical operation and sample and reagent consumption. Also, the time required by the entire process can be shortened. This work provides a simple and rapid screening tool for the investigation of cellular phosphorylation and signaling pathway for understanding the pathogenesis of cancer. In addition, the operation of the paper-based microreactor is compatible to the molecular biological training, and therefore, it has the potential to be developed for routine protocol for various research areas in conventional bioanalytical laboratories.
- Published
- 2014
38. Polyamidoamine starburst dendrimer-activated chromatography paper-based assay for sensitive detection of telomerase activity
- Author
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Zhenxin Wang, Zhen Lei, Hua Zhang, and Rongrong Tian
- Subjects
Paper ,Telomerase ,Dendrimers ,010402 general chemistry ,01 natural sciences ,Gene Expression Regulation, Enzymologic ,Analytical Chemistry ,HeLa ,Limit of Detection ,Dendrimer ,Cell Line, Tumor ,Polyamines ,Humans ,Enzyme Inhibitors ,Enzyme Assays ,Chromatography ,biology ,Chemistry ,010401 analytical chemistry ,Substrate (chemistry) ,biology.organism_classification ,0104 chemical sciences ,Paper chromatography ,Biochemistry ,Cell culture ,Cancer cell ,Primer (molecular biology) - Abstract
Telomerase is extensively expressed in various cancer cells and recognized as a target for cancer drug discovery. In the present study, a simple and amplification-free fluorescence assay based on polyamidoamine starburst dendrimer (PAMAM dendrimer)-activated paper device is proposed for sensitive detection of telomerase activity through hybridization of Cy5 modified single strand DNA probes with telomerase extension products. The paper substrate is fabricated by hand drawing according to a template, which is low cost, instrument free and easy operation. PAMAM is rich in amino groups on its surface and employed to immobilize the telomerase substrate (TS) primer. Highly sensitive detection of telomerase activity in HeLa cell lysate of 10 cells is achieved since the PAMAM dendrimer-activated paper surface can provide high density of binding sites for immobilization of TS primer. The experimental results also demonstrate that the assay can be employed to evaluate telomerase activity levels of various cell lines and screen telomerase inhibitors.
- Published
- 2017
39. In Situ Detection of Hydrogen Sulfide in 3D-Cultured, Live Prostate Cancer Cells Using a Paper-Integrated Analytical Device.
- Author
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Kim, Jae-Hyung, Lee, Young-Ju, Ahn, Yong-Jin, Kim, Minyoung, and Lee, Gi-Ja
- Subjects
CYSTEINE ,CANCER cells ,HYDROGEN sulfide ,PROSTATE cancer ,CELL culture ,CELL survival - Abstract
In this study, a paper-integrated analytical device that combined a paper-based colorimetric assay with a paper-based cell culture platform was developed for the in situ detection of hydrogen sulfide (H
2 S) in three-dimensional (3D)-cultured, live prostate cancer cells. Two kinds of paper substrates were fabricated using a simple wax-printing methodology to form the cell culture and detection zones, respectively. LNCaP cells were seeded directly on the paper substrate and grown in the paper-integrated analytical device. The cell viability and H2 S production of LNCaP cells were assessed using a simple water-soluble tetrazolium salt colorimetric assay and H2 S-sensing paper, respectively. The H2 S-sensing paper showed good sensitivity (sensitivity: 6.12 blue channel intensity/μM H2 S, R2 = 0.994) and a limit of quantification of 1.08 μM. As a result, we successfully measured changes in endogenous H2 S production in 3D-cultured, live LNCaP cells within the paper-integrated analytical device while varying the duration of incubation and substrate concentration (L-cysteine). This paper-integrated analytical device can provide a simple and effective method to investigate H2 S signaling pathways and drug screening in a 3D culture model. [ABSTRACT FROM AUTHOR]- Published
- 2022
- Full Text
- View/download PDF
40. Nanocomposite (Janus) paper as 3D cell culture system.
- Author
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Kehr, N.S. and Motealleh, A.
- Subjects
- *
NANOCOMPOSITE materials , *CELL culture , *CELL adhesion , *CELL differentiation , *AGGLOMERATION (Materials) , *THREE-dimensional imaging - Abstract
Nanocomposite (NC) papers using bifunctional nanoparticles are prepared for 3D cell growth experiments. Cells show better affinity to NC papers than to paper itself and differentiate the chemical group on the external surface of the nanoparticles. Cells possess spherical shaped morphology and form agglomerates onto the (NC) papers like as observed by cell growth in 3D materials. Furthermore, the preparation of “Janus” paper and the differentiation of its two distinct faces by cells is presented. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
41. Personalized Medicine: Engineered Paper-Based Cell Culture Platforms (Adv. Healthcare Mater. 22/2017)
- Author
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Darlin Lantigua, Barış Ünal, Gulden Camci-Unal, and Yan Ni Kelly
- Subjects
Engineering ,business.industry ,Biomedical Engineering ,Pharmaceutical Science ,Nanotechnology ,02 engineering and technology ,Paper based ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Biomaterials ,Tissue engineering ,Cell culture ,Engineering ethics ,Personalized medicine ,0210 nano-technology ,business - Published
- 2017
42. Towards a high throughput impedimetric screening of chemosensitivity of cancer cells suspended in hydrogel and cultured in a paper substrate
- Author
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Kin Fong Lei, Tai-Kun Liu, and Ngan-Ming Tsang
- Subjects
Paper ,Cell Survival ,Cell ,Biomedical Engineering ,Biophysics ,Cell Culture Techniques ,Antineoplastic Agents ,02 engineering and technology ,01 natural sciences ,Hydrogel, Polyethylene Glycol Dimethacrylate ,3D cell culture ,Cell Line, Tumor ,Neoplasms ,Electrochemistry ,medicine ,Electric Impedance ,Humans ,Doxorubicin ,Viability assay ,Etoposide ,Chemistry ,010401 analytical chemistry ,Substrate (chemistry) ,General Medicine ,Equipment Design ,Hep G2 Cells ,021001 nanoscience & nanotechnology ,Molecular biology ,0104 chemical sciences ,Cell biology ,High-Throughput Screening Assays ,medicine.anatomical_structure ,Cell culture ,Cancer cell ,Drug Screening Assays, Antitumor ,0210 nano-technology ,Biotechnology ,medicine.drug - Abstract
In order to achieve high predictive value of cell chemosensitivity test for clinical efficacy, cancer cells were suggested to be encapsulated and cultured in hydrogel to mimic the natural microenvironment of tumors. However, handling 3D cells/hydrogel culture construct is tedious and cellular response is difficult to be quantitatively analyzed. In the current study, a novel platform for conducting 3D cell culture and analyzing cell viability has been developed towards a high throughput drug screening. Cells encapsulated in the hydrogel were cultured in the microwells of a paper substrate. The substrate was then immersed in the culture medium containing drug for 2 days. At 24 and 48h during the culture course, the paper substrate was placed on the measurement electrodes for conducting the impedance measurement in order to quantify the cell viability in the hydrogel. Cell viability of two human hepatoma cell lines (Huh7 and Hep-G2) was quantitatively investigated under the treatment of two drugs (doxorubicin and etoposide). The results represented by IC50 values revealed that Huh7 cells had a higher drug resistance than Hep-G2 cells and doxorubicin had a higher efficacy than etoposide for treating hepatocellular carcinoma. The current work has demonstrated a high throughput, convenient, and quantitative platform for the investigation of chemosensitivity of cells cultured in the 3D environment.
- Published
- 2017
43. Toxicity of food contact paper evaluated by combined biological and chemical methods.
- Author
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Kejlová, Kristina, Dvořáková, Markéta, Vavrouš, Adam, Ševčík, Václav, Kanďárová, Helena, Letašiová, Silvia, Sosnovcová, Jitka, and Jírová, Dagmar
- Subjects
- *
FOOD toxicology , *HEALTH risk assessment , *ANDROGEN receptors , *ANALYTICAL chemistry , *CELL culture - Abstract
The study was focused on assessment of potential health risks of paper-based food contact materials (FCMs) in a step-wise approach using three toxicological bioassays in vitro and chemical analyses of migrating contaminants. 3T3 NRU cytotoxicity test showed high sensitivity to detect basal toxicity of FCMs extracts and served as a first-line test for selection of samples for further testing. The reconstructed human intestine model EpiIntestinal showed more realistic tissue response than cell culture monolayer and higher resistance despite prolonged exposure to the selected 6 samples, i.e. negligible decrease of viability and intestinal penetration, nevertheless an increase of IL-8 after exposure to black printed sample extract. Yeast based assays identified weak agonistic/antagonostic activity to human androgen receptor of the black printed sample. In accordance with the biological effects, the targeted LC and GC analytical methods confirmed the presence of high amounts of phthalates, photoinitiators and PAHs that could justify the hazard of the black printed sample. Heavily printed uncoated FCMs are recognized not to be suitable for direct contact with food. The selected bioassays and chemical analyses might be useful tools to detect targeted biological effects of xenobiotics suspected to contribute to human exposure from food. • An effect-directed approach combining bioassays and chemical analyses was proved as useful to assess possible impact of FCMs on human health. • bioassays included cytotoxicity, toxicity and penetration using reconstructed human intestinal model and endocrine disruption screen. • Extensively printed napkin samples exhibited highest toxicological risks and released phthalates, photoinitiators and PAHs. • The black printed paper extract caused IL-8 increase and agonistic activity to human androgen receptor. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
44. A Floating Capsule Electrochemical System for In Situ and Multichannel Ion-Selective Sensing.
- Author
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Yang, Jie, Ding, Ao, Zhou, Jia-Le, Yan, Bing-Yong, Gu, Zhen, and Wang, Hui-Feng
- Subjects
OXYGEN detectors ,ELECTROCHEMICAL sensors ,BIOTECHNOLOGICAL process monitoring ,CALCIUM ions ,CELL culture ,PHYSICAL measurements ,INDICATORS & test-papers ,FLOW velocity - Abstract
Free-floating electrochemical sensors are promising for in situ bioprocess monitoring with the advantages of movability, a lowered risk of contamination, and a simplified structure of the bioreactor. Although floating sensors were developed for the measurement of physical and chemical indicators such as temperature, velocity of flow, pH, and dissolved oxygen, it is the lack of available electrochemical sensors for the determination of the inorganic ions in bioreactors that has a significant influence on cell culture. In this study, a capsule-shaped electrochemical system (iCapsuleEC) is developed to monitor ions including K
+ , NH4 + , Na+ , Ca2+ , and Mg2+ based on solid-contact ion-selective electrodes (SC-ISEs). It consists of a disposable electrochemical sensor and signal-processing device with features including multichannel measurement, self-calibration, and wireless data transmission. The capacities of the iCapsuleEC were demonstrated not only for in situ measurement of ion concentrations but also for the optimization of the sensing electrodes. We also explored the possibility of the system for use in detection in simulated cell culture media. [ABSTRACT FROM AUTHOR]- Published
- 2023
- Full Text
- View/download PDF
45. Simultaneous Identification and Antimicrobial Susceptibility Testing of Multiple Uropathogens on a Microfluidic Chip with Paper-Supported Cell Culture Arrays
- Author
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Dayu Liu, Xiao-Xia Wen, Bin Chen, Banglao Xu, Jinqiong Lin, Bowen Shu, Guangtie Liang, Yan Du, and Mingyue Qi
- Subjects
Paper ,Staphylococcus aureus ,Cell Culture Techniques ,Antimicrobial susceptibility ,Nanotechnology ,02 engineering and technology ,Microbial Sensitivity Tests ,01 natural sciences ,Analytical Chemistry ,chemistry.chemical_compound ,Enterococcus faecalis ,Escherichia coli ,Humans ,Dimethylpolysiloxanes ,Chromatography ,Polydimethylsiloxane ,Chromogenic ,010401 analytical chemistry ,Microfluidic Analytical Techniques ,021001 nanoscience & nanotechnology ,Antimicrobial ,0104 chemical sciences ,Anti-Bacterial Agents ,Microfluidic chip ,chemistry ,Cell culture ,0210 nano-technology ,Urine sample - Abstract
A microfluidic chip was developed for one-step identification and antimicrobial susceptibility testing (AST) of multiple uropathogens. The polydimethylsiloxane (PDMS) microchip used had features of cell culture chamber arrays connected through a sample introduction channel. At the bottom of each chamber, a paper substrate preloaded with chromogenic media and antimicrobial agents was embedded. By integrating a hydrophobic membrane valve on the microchip, the urine sample can be equally distributed into and confined in individual chambers. The identification and AST assays on multiple uropathogens were performed by combining the spatial resolution of the cell culture arrays and the color resolution from the chromogenic reaction. The composite microbial testing assay was based on dynamic changes in color in a serial of chambers. The bacterial antimicrobial susceptibility was determined by the lowest concentration of an antimicrobial agent that is capable of inhibiting the chromogenic reaction. Using three common uropathogenic bacteria as test models, the developed microfluidic approach was demonstrated to be able to complete the multiple colorimetric assays in 15 h. The accuracy of the microchip method, in comparison with that of the conventional approach, showed a coincidence of 94.1%. Our data suggest this microfluidic approach will be a promising tool for simple and fast uropathogen testing in resource-limited settings.
- Published
- 2016
46. A Method to Efficiently Cryopreserve Mammalian Cells on Paper Platforms
- Author
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Muhammedin Deliorman, Roaa Alnemari, Mohammad A. Qasaimeh, and Pavithra Sukumar
- Subjects
Slow freezing ,Materials science ,Cryoprotectant ,Biocompatibility ,Dimethyl sulfoxide ,Strategy and Management ,Mechanical Engineering ,Metals and Alloys ,Spheroid ,Industrial and Manufacturing Engineering ,Cryopreservation ,chemistry.chemical_compound ,chemistry ,Cell culture ,Methods Article ,Fiber ,Biological system - Abstract
This protocol describes a simple method to cryopreserve mammalian cells within filter papers as an alternative to conventional slow-freezing approach. The method involves treating paper fibers with fibronectin, using low concentrations of the cryoprotectant dimethyl sulfoxide (DMSO), and slow freezing cells to -80 °C at a 1 °C min(-1) rate. In our method, the biocompatibility, large surface area, 3D porosity and fiber flexibility of the paper, in combination with the fibronectin treatment, yield recovery of cells comparable to conventional approaches, with no additional fine-tuning to freezing and thawing procedures. We expect that the paper-based cryopreservation method will bring several advantages to the field of preserving mammalian cells, including accommodation of a higher number of cells within a unit volume and no cell loss after release. The method requires a minimal storage space, where paper platforms with large areas can be rolled and/or folded and stored in stocks, and allows for efficient transportation/distribution of cells in an on-demand manner. Moreover, an additional feature of this method includes the formation and cryopreservation of cellular spheroids and 3D cell cultures.
- Published
- 2020
47. Integrated paper-based 3D platform for long-term cell culture and in situ cell viability monitoring of Alzheimer's disease cell model
- Author
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Yu Zhong, Li Shanhong, Xinhua Lin, Zi-Zhen Guo, Yao Chen, Ai-Lin Liu, Wei Chen, Meng-Meng Liu, Yun Lei, and Hui Liu
- Subjects
Amyloid beta ,Cell Survival ,Cell Culture Techniques ,Apoptosis ,02 engineering and technology ,01 natural sciences ,PC12 Cells ,Analytical Chemistry ,chemistry.chemical_compound ,Alzheimer Disease ,Animals ,Viability assay ,Hydrogen peroxide ,chemistry.chemical_classification ,Reactive oxygen species ,Amyloid beta-Peptides ,biology ,Superoxide ,010401 analytical chemistry ,Reproducibility of Results ,Hydrogen Peroxide ,021001 nanoscience & nanotechnology ,Cytoprotection ,Peptide Fragments ,0104 chemical sciences ,Rats ,chemistry ,Cell culture ,Curcumin ,Biophysics ,biology.protein ,0210 nano-technology - Abstract
Reactive oxygen species including superoxide anion, hydrogen peroxide (H2O2) and hydroxyl radicals, as a conflicting class of biological metabolites in living organism, act crucial effect on Alzheimer's disease (AD). In this work, a facile integrated platform composed of a paper-based three-dimension (3D) cell culture system and an electrochemical sensor was developed for the construction of AD cell model in third dimensional structure and in situ cell viability monitoring by H2O2 released from PC12 cells cultured on paper scaffold were divided into three groups containing control group, amyloid beta peptide 25–35 (Aβ25-35) group and Aβ25-35+curcumin (Aβ25-35+cur) group, respectively. In addition, the paper-based 3D platform displayed excellent properties, such as sensitivity, selectivity, reproducibility and stability. The levels of H2O2 expressed in PC12 cells of the three groups were monitored through a paper-based 3D platform. The viability of cells cultured on the 96-well plate was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Results of this paper-based platform are consistent with those of MTT, both displaying improved cell viability and decreased H2O2 production in Aβ25-35+cur group compared to Aβ25-35 group, which indicates that curcumin has effective cytoprotection. The paper-based 3D platform provides a convenient, economic and universal platform for in situ cell activity monitoring by key small molecules released from living cells.
- Published
- 2020
48. Low magnitude high frequency vibrations expedite the osteogenesis of bone marrow stem cells on paper based 3D scaffolds
- Author
-
Engin Ozcivici, Gulistan Mese, and Ozge Karadas
- Subjects
biology ,Chemistry ,0206 medical engineering ,Mesenchymal stem cell ,Biomedical Engineering ,Bone Marrow Stem Cell ,02 engineering and technology ,Bone tissue ,020601 biomedical engineering ,01 natural sciences ,010309 optics ,Extracellular matrix ,medicine.anatomical_structure ,Cell culture ,0103 physical sciences ,medicine ,Biophysics ,Osteocalcin ,biology.protein ,Alkaline phosphatase ,Original Article ,Stem cell - Abstract
Anabolic effects of low magnitude high frequency (LMHF) vibrations on bone tissue were consistently shown in the literature in vivo, however in vitro efforts to elucidate underlying mechanisms are generally limited to 2D cell culture studies. Three dimensional cell culture platforms better mimic the natural microenvironment and biological processes usually differ in 3D compared to 2D culture. In this study, we used laboratory grade filter paper as a scaffold material for studying the effects of LHMF vibrations on osteogenesis of bone marrow mesenchymal stem cells in a 3D system. LMHF vibrations were applied 15 min/day at 0.1 g acceleration and 90 Hz frequency for 21 days to residing cells under quiescent and osteogenic conditions. mRNA expression analysis was performed for alkaline phosphatase (ALP) and osteocalcin (OCN) genes, Alizarin red S staining was performed for mineral nodule formation and infrared spectroscopy was performed for determination of extracellular matrix composition. The highest osteocalcin expression, mineral nodule formation and the phosphate bands arising from the inorganic phase was observed for the cells incubated in osteogenic induction medium with vibration. Our results showed that filter paper can be used as a model scaffold system for studying the effects of mechanical loads on cells, and LMHF vibrations induced the osteogenic differentiation of stem cells.
- Published
- 2020
49. Paper as a scaffold for cell cultures: Teaching an old material new tricks
- Author
-
Kierra Walsh, Sanika Suvarnapathaki, Gulden Camci-Unal, and Xinchen Wu
- Subjects
0301 basic medicine ,Scaffold ,Materials science ,Nanotechnology ,02 engineering and technology ,021001 nanoscience & nanotechnology ,3d shapes ,Biocompatible material ,03 medical and health sciences ,030104 developmental biology ,Tissue engineering ,Cell culture ,General Materials Science ,0210 nano-technology - Abstract
Paper-based cell culture platforms have emerged as a promising approach for a myriad of biomedical applications, such as tissue engineering, disease models, cancer research, biotechnology, high-throughput testing, biosensing, and diagnostics. Paper enables the generation of highly flexible, biocompatible, inexpensive, porous, and three-dimensional (3D) constructs and devices. These systems have been used to culture mammalian cells, bacteria, algae, and fungi. Studies have shown that paper is an exceptional material for applications in life sciences, materials sciences, engineering, and medicine. Paper has been employed for creating biomimetic cell culture environments by folding or stacking it into the desired 3D shapes and structures. This review discusses the use of paper-based platforms for cellular applications and provides a diverse range of examples.
- Published
- 2018
50. Biocompatibility evaluation of cigarette and carbon papers used in repair of traumatic tympanic membrane perforations: experimental study
- Author
-
Zeynep Sümer, Emine Elif Altuntaş, and [Altuntas, Emine Elif] Cumhuriyet Univ, Dept Otorhinolaryngol, Sch Med, TR-58140 Sivas, Turkey -- [Altuntas, Emine Elif] Univ Cumhuriyet, Dept Ear Nose & Throat Educ, Sivas, Turkey -- [Altuntas, Emine Elif] Univ Cumhuriyet, Fac Med, Res Hosp, Sivas, Turkey -- [Sumer, Zeynep] Univ Cumhuriyet, Dept Microbiol, Fac Med, Sivas, Turkey
- Subjects
Paper ,medicine.medical_specialty ,Turkey ,Biocompatibility ,Scanning electron microscope ,Tympanic membrane ,Tetrazolium Salts ,Biocompatible Materials ,MTT test ,In Vitro Techniques ,Statistics, Nonparametric ,Mice ,Cell Adhesion ,medicine ,Animals ,Cell adhesion ,Cytotoxicity ,Cell Proliferation ,Paper patches ,Tympanic Membrane Perforation ,Cell growth ,business.industry ,Traumatic perforation ,General Medicine ,Fibroblasts ,Carbon ,In vitro ,Surgery ,Thiazoles ,Membrane ,Otorhinolaryngology ,Cell culture ,Microscopy, Electron, Scanning ,business ,Scanning electron microscopy ,Biomedical engineering - Abstract
WOS: 000313019700012, PubMed ID: 22271281, The purposes of this study were to investigate the biocompatibility of two different paper patches (carbon and cigarette papers) and compare the adhesion and proliferation features of L929 fibroblast cells by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT Test) test and scanning electron microscopy (SEM). In this study, time-dependent cytotoxic effects of cigarette and carbon papers used in repairing small traumatic TM perforations were investigated in vitro by using MTT test. And also adhesion and spreading of cells over disk surface were observed by SEM. Cytotoxicity test carried out by MTT analysis on leakage products collected from two types of paper patches at the end of 24 and 48 h revealed no cytotoxicity (P > 0.05). In SEM studies, it was observed that cells started to proliferate over disk surface as a result of 48-h incubation, and SEM revealed that the cell proliferation over cigarette paper was more compared to the one over carbon paper. We believe that this is the first study where biocompatibility and adhesion features of carbon and cigarette paper have been studied by using L929 fibroblast cell culture. As a result, biocompatibility of cigarette paper and also whether cigarette paper was superior to carbon paper in cell attachment and biocompatibility were studied. It was found, by MTT test and SEM test, that cigarette paper had a higher biocompatibility and cell attachment, and thus cigarette paper should be the patch to be preferred in cases where TM perforations are repaired by paper-patch method.
- Published
- 2012
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