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Introduction: Metformin (MET), an old anti-diabetic drug, has proven unexpected anti-glioblastoma effects, by impacting cell proliferation, migration and invasion. However, its remarkable anti-cancer efficacy is mainly limited to the use of high millimolar concentrations in in vitro studies, which are hard to be attained in the clinical setting. Aim: The aim of this paper was to synthetize gold nanoparticles loaded with MET and to test if an enhanced drug delivery via nanotechnology could overcome the limitations of small drug concentrations. Materials and Methods: Gold nanoparticles were functionalized with chitosan (GNPc) and loaded with 80 µM of MET. Their size, zeta potential and stability were characterized and their internalization within tumor cells was assayed through dark field microscopy. Three primary glioblastoma stem cell lines were treated with 5, 10 and 20 µg/mL concentrations of nanoparticles and irradiated. The anti-tumoral effect was evaluated through the MTT cell viability assay. Results: MET-GNPc are easily synthetized and have a positive zeta potential, spherical shape and a median size of 26 nm. MET-GNPc have an increased cell internalization and affect the viability of all three glioblastoma cell lines used compared to control and free MET. However, their anti-cancer effect is not statistically different when compared to GNPc, although a slight tendency to a better response may be observed. Conclusion:Despite an increased cell internalization, the small micromolar concentrations of metformin does not bring an additional benefit to chitosan-based GNPs. Novel delivery methods being able to carry a higher drug concentration of metformin should be tested.

Bone marrow stromal cells (MSCs) represent a heterogeneous population derived from the non–blood-forming fraction of bone marrow that regulates hematopoietic cell development. In vitro, adult mesenchymal stem cells resident in this bone marrow fraction differentiate into bone, cartilage, and fat. Because MSCs can be easily obtained using a simple bone marrow aspiration and show extensive capacity for expansion in vitro, these cells have been considered as candidates for cell therapy. The aim of this study was to purify rat MSCs from adult bone marrow and to functionally characterise their abilities to differentiate along diverse lineages. Our data demonstrate that we successfully isolated, culture-expanded and differentiated a relatively homogeneous population of MPCs from adult rat bone marrow.

The antitumoral, antioxidant, and anti-inflammatory effects of flaxseed ethanol extract was screened. Phytochemical analysis was performed by measuring the total phenolic content and by HPLC-DAD-ESI MS. In vitro antiproliferative activity was appreciated by MMT test of four adenocarcinomas and two normal cell lines. In vitro, antioxidant activity was evaluated by DPPH, FRAP, H2O2, and NO scavenging tests. The in vivo growth inhibitory activity against Ehrlich ascites carcinoma (EAC) in female BALB/c mice was determined using the trypan blue test. In EAC mice serum and ascites total oxidative status, total antioxidant reactivity, oxidative stress index, malondialdehyde, total thiols, total nitrites, 3-nitrotyrosine, and NFkB were measured. The phytochemical analysis found an significant content of phenols, with lignans having the highest concentration. The extract had an significant in vitro antioxidant effect and different inhibitory effects on different cell lines. After treatment of EAC mice with flaxseeds extract, body weight, ascites volume and viable tumour cell count, serum and ascites oxidative stress, and inflammatory markers decreased significantly. The ethanol flaxseeds extract has potential antiproliferative activity against some ovary and endometrial malignant cells and EAC. This effect can be attributed to the phenols content, and its antioxidant and anti-inflammatory activity.

Inflammation is a major pathogenic pathway in pulmonary chronic obstructive disease (COPD). Interleukin-6 (IL-6) mediates the local and systemic immune response. The aim consisted in investigating the relationship between IL-6 serum levels and IL-6 -597A/G gene polymorphism (rs1800797) with COPD. Serum levels of IL-6 were determined using an enzyme-linked immune-sorbent assay, in 120 participants (60 COPD patients and 60 healthy subjects), from Transylvanian region. The IL-6 -597A/G gene polymorphism was investigated by high molecular weight genomic DNA extracted from the peripheral blood leukocytes, and subsequently analyzed by the Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) technique. Smoking history, the severity of the disease, expressed by the GOLD stages, and arterial blood partial pressure of oxygen (PaO 2 ) levels were also investigated. COPD patients had significantly elevated blood levels of IL-6 when compared to the control group ( p

Background and objective: The aim of the present study was to establish a new differentiation protocol using cannabidiol (CBD) and vitamin D3 (Vit. D3) for a better and faster osteogenic differentiation of dental tissue derived mesenchymal stem cells (MSCs). Materials and methods: MSCs were harvested from dental follicle (DFSCs), dental pulp (DPSCs), and apical papilla (APSCs) of an impacted third molar of a 17-year old patient. The stem cells were isolated and characterized using flow cytometry; reverse transcription polymerase chain reaction (RT-PCR); and osteogenic, chondrogenic, and adipogenic differentiation. The effects of CBD and Vit. D3 on osteogenic differentiation of dental-derived stem cell were evaluated in terms of viability/metabolic activity by alamar test, expression of collagen1A, osteopontin (OP), osteocalcin (OC), and osteonectin genes and by quantification of calcium deposits by alizarin red assay. Results: Stem cell characterization revealed more typical stemness characteristics for DFSCs and DPSCs and atypical morphology and markers expression for APSCs, a phenotype that was confirmed by differences in multipotential ability. The RT-PCR quantification of bone matrix proteins expression revealed a different behavior for each cell type, APSCs having the best response for CBD. DPSCs showed the best osteogenic potential when treated with Vit. D3. Cultivation of DFSC in standard stem cell conditions induced the highest expression of osteogenic genes, suggesting the spontaneous differentiation capacity of these cells. Regarding mineralization, alizarin red assay indicated that DFSCs and APSCs were the most responsive to low doses of CBD and Vit. D3. DPSCs had the lowest mineralization levels, with a slightly better response to Vit. D3. Conclusions: This study provides evidence that DFSCs, DPSCs, and APSCs respond differently to osteoinduction stimuli and that CBD and Vit. D3 can enhance osteogenic differentiation of these types of cells under certain conditions and doses.

Background: SLM (Selective Laser Melting)–manufactured Titanium (Ti) scaffolds have a significant value for bone reconstructions in the oral and maxillofacial surgery field. While their mechanical properties and biocompatibility have been analysed, there is still no adequate information regarding tissue integration. Therefore, the aim of this study is a comprehensive systematic assessment of the essential parameters (porosity, pore dimension, surface treatment, shape) required to provide the long-term performance of Ti SLM medical implants. Materials and methods: A systematic literature search was conducted via electronic databases PubMed, Medline and Cochrane, using a selection of relevant search MeSH terms. The literature review was conducted using the preferred reporting items for systematic reviews and meta-analysis (PRISMA). Results: Within the total of 11 in vitro design studies, 9 in vivo studies, and 4 that had both in vitro and in vivo designs, the results indicated that SLM-generated Ti scaffolds presented no cytotoxicity, their tissue integration being assured by pore dimensions of 400 to 600 µm, high porosity (75–88%), hydroxyapatite or SiO2–TiO2 coating, and bioactive treatment. The shape of the scaffold did not seem to have significant importance. Conclusions: The SLM technique used to fabricate the implants offers exceptional control over the structure of the base. It is anticipated that with this technique, and a better understanding of the physical interaction between the scaffold and bone tissue, porous bases can be tailored to optimize the graft’s integrative and mechanical properties in order to obtain structures able to sustain osseous tissue on Ti.

Despite recent advances in disease management and prevention, heart failure (HF) prevalence is still high. Hypertension, inflammation and oxidative stress are being investigated as important causative processes in HF. L. barbarum L. polysaccharides (LBPs) are widely used for their anti-inflammatory and antioxidant properties. Thus, the aim of the present study was to evaluate the effects of LBPs on inflammation and oxidative stress markers in a pressure overload-induced HF rat model, surgically induced by abdominal aorta banding in Wistar rats (AAB) (n = 28). Also, control rats (n = 10) were subjected to a sham operation. After echocardiographic confirmation of HF (week 24), AAB rats were divided into three groups: rats treated with LBPs for 12 weeks: 100 mg/kg body weight /day (AAB_100, n = 9), 200 mg/kg body weight /day (AAB_200, n = 7) and no-treatment group (control AAB, n = 12). After 12 weeks of treatment with LBPs, the decline of cardiac function was prevented compared to the control AAB rats. Treatment with 200 mg/kg body weight /day LBPs significantly reduced the inflammation as seen by cytokine levels (IL-6 and TNF-α) and the plasma lipid peroxidation, as seen by malondialdehyde levels. These results suggest that LBPs present anti-inflammatory and antioxidant effects with utility in a HF animal model and encourage further investigation of the cardioprotective effects of these polysaccharides.

Currently, there is no cure for the permanent vision loss caused by degenerative retinal diseases. One of the novel therapeutic strategies aims at the development of stem cells (SCs) based neuroprotective and regenerative medicine. The main sources of SCs for the treatment of retinal diseases are the embryo, the bone marrow, the region of neuronal genesis, and the eye. The success of transplantation depends on the origin of cells, the route of administration, the local microenvironment, and the proper combinative formula of growth factors. The feasibility of SCs based therapies for degenerative retinal diseases was proved in the preclinical setting. However, their translation into the clinical realm is limited by various factors: the immunogenicity of the cells, the stability of the cell phenotype, the predilection of SCs to form tumors in situ, the abnormality of the microenvironment, and the association of a synaptic rewiring. To improve SCs based therapies, nanotechnology offers a smart delivery system for biomolecules, such as growth factors for SCs implantation and differentiation into retinal progenitors. This review explores the main advances in the field of retinal transplantology and applications of nanotechnology in the treatment of retinal diseases, discusses the challenges, and suggests new therapeutic approaches in retinal transplantation.

GlioStat (patent pending) denotes an invention that addresses the current deficiencies of glioblastoma (GBM) chemotherapy by developing a molecularly imprinted drug reservoir that is specifically designed for post-surgical recovery. The goal of our research was to guarantee the long-term release of the antitumor drug ruxolitinib (RUX), which is specifically designed to target residual infiltrative cancer cells, while simultaneously reducing the risk of adverse effects. In order to achieve this goal, we have successfully developed and characterized four distinctive molecularly imprinted polymers (MIPs), with one of them advancing to in vivo experimentation. The selection of one MIP for further in vivo investigations was guided by the Alamar Blue viability assay on C6 GBM cell cultures, which took into account both the efficacy and the potential toxicity of residual monomers. Over the course of 96 hours, MIP 2 demonstrated the most favorable risk-benefit profile, providing superior efficacy against GBM C6 cells. In contrast, its non-imprinted counterpart (NIP 2) exhibited minimal toxicity. The protocol involved anesthetising the male Wistar rats, immobilising them on a corkboard, without the use of a stereotactic headholder. After shaving the head and local disinfection with iodine solution, we made a linear sagittal incision and exposed the parietal bones. A 3x3 mm burr hole was made with a pneumatic drill in the right parietal bone, revealing the dura mater. Subsequently, we injected 20,000 C6 GBM cells suspended in 5 microliters of solution in the cerebral parenchyma at a depth of 3 mm. Clinical and imaging control was performed. For the animals that developed tumors, a second therapeutic intervention was performed. The rats were anesthetised, disinfected, and readied in the same manner as before. The burr hole was exposed, with the tumor being partially resected via a “microscooping” technique. Next, 5-10 microliters of MIP2 solution (4 mg/mL) were injected into the parenchyma respective of the tumor bed. Five microliters of thrombin solution were then added (100 UI/mL) to form the fibrin network. In comparison to the untreated controls and animals treated with free RUX, animals treated with MIP2 exhibited a substantial increase in survival time during the in vivo evaluation, extending from 20 to 50 days. As such, our research had yielded a potentially life-changing drug delivery system in GBM, with the capacity to significantly increase postoperative survival. More in vivo tests should be conducted to validate our findings.

Abstract Background Cancer stem-like cells (CSCs) have been extensively researched as the primary drivers of therapy resistance and tumor relapse in patients with breast cancer. However, due to lack of specific molecular markers, increased phenotypic plasticity and no clear clinicopathological features, the assessment of CSCs presence and functionality in solid tumors is challenging. While several potential markers, such as CD24/CD44, have been proposed, the extent to which they truly represent the stem cell potential of tumors or merely provide static snapshots is still a subject of controversy. Recent studies have highlighted the crucial role of the tumor microenvironment (TME) in influencing the CSC phenotype in breast cancer. The interplay between the tumor and TME induces significant changes in the cancer cell phenotype, leading to the acquisition of CSC characteristics, therapeutic resistance, and metastatic spread. Simultaneously, CSCs actively shape their microenvironment by evading immune surveillance and attracting stromal cells that support tumor progression. Methods In this study, we associated in vitro mammosphere formation assays with bulk tumor microarray profiling and deconvolution algorithms to map CSC functionality and the microenvironmental landscape in a large cohort of 125 breast tumors. Results We found that the TME score was a significant factor associated with CSC functionality. CSC-rich tumors were characterized by an immune-suppressed TME, while tumors devoid of CSC potential exhibited high immune infiltration and activation of pathways involved in the immune response. Gene expression analysis revealed IFNG, CXCR5, CD40LG, TBX21 and IL2RG to be associated with the CSC phenotype and also displayed prognostic value for patients with breast cancer. Conclusion These results suggest that the characterization of CSCs content and functionality in tumors can be used as an attractive strategy to fine-tune treatments and guide clinical decisions to improve patients therapy response.

Many preclinical studies have shown that birth-associated tissues, cells and their secreted factors, otherwise known as perinatal derivatives (PnD), possess various biological properties that make them suitable therapeutic candidates for the treatment of numerous pathological conditions. Nevertheless, in the field of PnD research, there is a lack of critical evaluation of the PnD standardization process: from preparation to in vitro testing, an issue that may ultimately delay clinical translation. In this paper, we present the PnD e-questionnaire developed to assess the current state of the art of methods used in the published literature for the procurement, isolation, culturing preservation and characterization of PnD in vitro. Furthermore, we also propose a consensus for the scientific community on the minimal criteria that should be reported to facilitate standardization, reproducibility and transparency of data in PnD research. Lastly, based on the data from the PnD e-questionnaire, we recommend to provide adequate information on the characterization of the PnD. The PnD e-questionnaire is now freely available to the scientific community in order to guide researchers on the minimal criteria that should be clearly reported in their manuscripts. This review is a collaborative effort from the COST SPRINT action (CA17116), which aims to guide future research to facilitate the translation of basic research findings on PnD into clinical practice.

The potential clinical applications of human amniotic membrane (hAM) and human amniotic epithelial cells (hAECs) in the field of regenerative medicine have been known in literature since long. However, it has yet to be elucidated whether hAM contains different anatomical regions with different plasticity and differentiation potential. Recently, for the first time, we highlighted many differences in terms of morphology, marker expression, and differentiation capabilities among four distinct anatomical regions of hAM, demonstrating peculiar functional features in hAEC populations. The aim of this study was to investigate in situ the ultrastructure of the four different regions of hAM by means of transmission electron microscopy (TEM) to deeply understand their peculiar characteristics and to investigate the presence and localization of secretory products because to our knowledge, there are no similar studies in the literature. The results of this study confirm our previous observations of hAM heterogeneity and highlight for the first time that hAM can produce extracellular vesicles (EVs) in a heterogeneous manner. These findings should be considered to increase efficiency of hAM applications within a therapeutic context.

This work aimed to compare the effect of four new toothpastes (P1–P4) based on pure and biomimetic substituted nano-hydroxyapatites (HAPs) on remineralization of human enamel. Artificially demineralized enamel slices were daily treated for ten days with different toothpastes according to the experimental design. Tooth enamel surfaces were investigated using atomic force microscope (AFM) images and surface roughness (Ra) determined before and after treatment. The surface roughness of enamel slices was statistically analyzed by one-way ANOVA and Bonferroni’s multiple comparison test. X-ray diffraction (XRD) and Fourier transform infrared (FTIR) data revealed the HAP structure with crystal sizes between 28 and 33 nm and crystallinity between 29 and 37%. The average size of HAP particles was found to be between 30 and 40 nm. The Ra values indicated that P3 (HAP-Mg-Zn-Sr-Si) toothpaste was the most effective after 10 days of treatment, leading to the lowest mean roughness. The P3 and P2 (HAP) toothpastes were found to be effective in promoting remineralization. Specifically, their effectiveness can be ranked as follows: P3 = P2 > P4 (HAP-Mg-Zn-Si) > P1 (HAP-Zn), considering both the chemical composition and the size of their constitutive nanoparticles. The proposed toothpastes might be used successfully to treat early tooth decay.

In women, breast cancer is the most commonly diagnosed cancer (11.7% of total cases) and the leading cause of cancer death (6.9%) worldwide. Bioactive dietary components such as Sea buckthorn berries are known for their high carotenoid content, which has been shown to possess anti-cancer properties. Considering the limited number of studies investigating the bioactive properties of carotenoids in breast cancer, the aim of this study was to investigate the antiproliferative, antioxidant, and proapoptotic properties of saponified lipophilic Sea buckthorn berries extract (LSBE) in two breast cancer cell lines with different phenotypes: T47D (ER+, PR+, HER2−) and BT-549 (ER-, PR-, HER2−). The antiproliferative effects of LSBE were evaluated by an Alamar Blue assay, the extracellular antioxidant capacity was evaluated through DPPH, ABTS, and FRAP assays, the intracellular antioxidant capacity was evaluated through a DCFDA assay, and the apoptosis rate was assessed by flow cytometry. LSBE inhibited the proliferation of breast cancer cells in a concentration-dependent manner, with a mean IC50 of 16 µM. LSBE has proven to be a good antioxidant both at the intracellular level, due to its ability to significantly decrease the ROS levels in both cell lines (p = 0.0279 for T47D, and p = 0.0188 for BT-549), and at the extracellular level, where the ABTS and DPPH inhibition vried between 3.38–56.8%, respectively 5.68–68.65%, and 35.6 mg/L equivalent ascorbic acid/g LSBE were recorded. Based on the results from the antioxidant assays, LSBE was found to have good antioxidant activity due to its rich carotenoid content. The flow cytometry results revealed that LSBE treatment induced significant alterations in late-stage apoptotic cells represented by 80.29% of T47D cells (p = 0.0119), and 40.6% of BT-549 cells (p = 0.0137). Considering the antiproliferative, antioxidant, and proapoptotic properties of the carotenoids from LSBE on breast cancer cells, further studies should investigate whether these bioactive dietary compounds could be used as nutraceuticals in breast cancer therapy.

Progress in the understanding of the biology of perinatal tissues has contributed to the breakthrough revelation of the therapeutic effects of perinatal derivatives (PnD), namely birth-associated tissues, cells, and secreted factors. The significant knowledge acquired in the past two decades, along with the increasing interest in perinatal derivatives, fuels an urgent need for the precise identification of PnD and the establishment of updated consensus criteria policies for their characterization. The aim of this review is not to go into detail on preclinical or clinical trials, but rather we address specific issues that are relevant for the definition/characterization of perinatal cells, starting from an understanding of the development of the human placenta, its structure, and the different cell populations that can be isolated from the different perinatal tissues. We describe where the cells are located within the placenta and their cell morphology and phenotype. We also propose nomenclature for the cell populations and derivatives discussed herein. This review is a joint effort from the COST SPRINT Action (CA17116), which broadly aims at approaching consensus for different aspects of PnD research, such as providing inputs for future standards for the processing and in vitro characterization and clinical application of PnD.

Glioblastoma multiforme (GBM) is one of the deadliest brain tumors. Current standard therapy includes tumor resection surgery followed by radiotherapy and chemotherapy. Due to the tumors invasive nature, recurrences are almost a certainty, giving the patients after diagnosis only a 12–15 months average survival time. Therefore, there is a dire need of finding new therapies that could potentially improve patient outcomes. Ferroptosis is a newly described form of cell death with several implications in cancer, among which GBM. Agents that target different molecules involved in ferroptosis and that stimulate this process have been described as potentially adjuvant anti-cancer treatment options. In GBM, ferroptosis stimulation inhibits tumor growth, improves patient survival, and increases the efficacy of radiation and chemotherapy. This review provides an overview of the current knowledge regarding ferroptosis modulation in GBM.

Bone regeneration is a central focus of maxillofacial research, especially when dealing with dental implants or critical sized wound sites. While bone has great regeneration potential, exogenous delivery of growth factors can greatly enhance the speed, duration, and quality of osseointegration, making a difference in a patient’s quality of life. Bone morphogenic protein 2 (BMP-2) is a highly potent growth factor that acts as a recruiting molecule for mesenchymal stromal cells, induces a rapid differentiation of them into osteoblasts, while also maintaining their viability. Currently, the literature data shows that the liposomal direct delivery or transfection of plasmids containing BMP-2 at the bone wound site often results in the overexpression of osteogenic markers and result in enhanced mineralization with formation of new bone matrix. We reviewed the literature on the scientific data regarding BMP-2 delivery with the help of liposomes. This may provide the ground for a future new bone regeneration strategy with real chances of reaching clinical practice.

Circulating tumor cells (CTCs) are nowadays one of the most promising tumor biomarkers. It is well correlated with overall survival and progression-free survival in breast cancer, as well as in many other types of human cancer. In addition, enumeration and analysis of CTCs could be important for monitoring the response to different therapeutic agents, thus guiding the treatment of cancer patients and offering the promise of a more personalized approach. In this article, we present a new method that could be used for the automatic detection of CTC in blood, based on the microscopic appearance of unstained cells. The proposed method is based on the evaluation of image characteristics and boosting techniques. A dataset of 263 dark field microscopy images was constructed and used for our tests, containing blood spiked with three different types of tumor cells. An overall sensitivity of 92.87% and a specificity of 99.98% were obtained for the detection of CTC, performances which proved to be comparable to those obtained by human experts.