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In this contribution, a novel, low-cost, high throughput, and ultra-selective electrochemical DNA nanobiosensor was developed for accurate on-site detection of Mycobacterium avium subspecies paratuberculosis (MAP) in real media for practical diagnosis of Johne's disease (JD). The method was designed based on the immobilization of graphene oxide and chitosan biopolymer on the surface of a glassy carbon electrode, modified by electrochemical immobilization of graphene oxide and chitosan biopolymer, followed by activation of biopolymer via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxy succinimide (EDC/NHS) coupling system. Afterward, the commercial probe DNA (ssDNA) was stabilized on the activated electrode surface to prepare an ultra-selective ssDNA-stabilized nanobiosensor for MAP sensing called "ssDNA-stabilized GO-CH-EDC/NHS-modified electrode". Several characterization methods distinguished the bioelectrode. The DNA hybridization between the nanobiosensor and target DNA was confirmed by cyclic voltammetry and differential pulse voltammetry. "At optimal experimental conditions, the nanobiosensor showed a linear range of 1.0 × 10 -15 -1.0 × 10 -12  mol L -1 , a detection limit as low as 1.53 × 10 -13  mol L -1 , and a repeatability with a relative standard deviation (%RSD) of 4.7%. The reproducibility was also appropriate, with a %RSD of about 10%. It was used to diagnose MAP in real samples with highly accurate results. Therefore, the developed nanobiosensor can be used for clinical diagnosis of MAP., (© 2024. The Author(s).)

Nanoparticles (NPs) based therapies for Alzheimer's disease (AD) attract interest due to their ability to pass across or bypass the blood-brain barrier. Chitosan (CS) NPs or graphene quantum dots (GQDs) are promising drug carriers with excellent physicochemical and electrical properties. The current study proposes the combination of CS and GQDs in ultrasmall NP form not as drug carriers but as theranostic agents for AD. The microfluidic-based synthesis of the CS/GQD NPs with optimized characteristics makes them ideal for transcellular transfer and brain targeting after intranasal (IN) delivery. The NPs have the ability to enter the cytoplasm of C6 glioma cells in vitro and show dose and time-dependent effects on the viability of the cells. IN administration of the NPs to streptozotocin (STZ) induced AD-like models lead to a significant number of entrances of the treated rats to the target arm in the radial arm water maze (RAWM) test. It shows the positive effect of the NPs on the memory recovery of the treated rats. The NPs are detectable in the brain via in vivo bioimaging due to GQDs as diagnostic markers. The noncytotoxic NPs localize in the myelinated axons of hippocampal neurons. They do not affect the clearance of amyloid β (Aβ) plaques at intercellular space. Moreover, they showed no positive impact on the enhancement of MAP2 and NeuN expression as markers of neural regeneration. The memory improvement in treated AD rats may be due to neuroprotection via the anti-inflammation effect and regulation of the brain tissue microenvironment that needs to be studied., (© 2023 Wiley-VCH GmbH.)

Zoledronic acid (ZA) is known as a potent bisphosphonate in osteogenic differentiation, but at high doses, it possesses toxic effects and causes decreased proliferation and differentiation of osteoblasts. Therefore, encapsulation of ZA into nanoparticles and control of its release is expected to promote differentiation of stem cells into osteoblasts. The present work aimed to develop a simple method for synthesis of monodisperse ZA-loaded chitosan (CS) nanoparticles. In this regard, we proposed a microfluidic synthesis of nanoparticles through the ionic cross-linking of CS in the presence of ZA without a crosslinker. The main advantages of these microfluidic generated nanoparticles were narrow size distribution and fine spherical shape. Conversely, the nanoparticles that were synthesized using a bulk mixing method had an irregular shape with a broad size distribution. Real-time PCR assay as well as alizarin red staining were used to evaluate the in-vitro osteogenic potential of the nanoparticles. The results indicated that the controlled release of ZA from the microfluidic system generated uniform nanoparticles, improving the osteogenic differentiation of mesenchymal stem cells. Additionally, this microfluidic device provided the well-controlled synthesis of novel nanoparticles with a modified CS macromolecular polymer for targeted drug delivery systems., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2022. Published by Elsevier B.V.)

The present study evaluated the potential of nanochitosan coating enriched with ethanol-water extract of the walnut green husk (WGHE) on spoilage and rancidity of rainbow trout (Oncorhynchus mykiss) during six-day refrigerated storage. Hence, we have considered fresh trout fillets without any treatment as control (C), immersed in 2% solution of chitosan nanoparticles (CN), combination with 1.5% and 3% WGHE with nanochitosan coating (CN + WGHE 1.5 and CN + WGHE3), for physicochemical, microbial and sensorial assays. The highest levels and total volatile nitrogen were observed after day 6 in C, while the lowest was found in CN + WGHE3 groups. Thiobarbituric acid reactive substance (TBARS) and peroxide value of untreated fillets on day 6 of the study were significantly higher than NC + WGHE3 with 0.08 mg/g and 3.27 mEq/kg, respectively. The total microbial population was: C ˃ CN ˃ CN + WGHE 1.5 > CN + WGHE3, which expresses the effect of the extract on the total microbial population. Overall, the combination of WGHE with CN increased the extract's efficiency in reducing peroxide value, TBARS, and total volatile nitrogen and delayed the pH increase, improving the overall acceptability of rainbow trout fillets stored in refrigerated conditions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Aims: Progenitor cells-based regenerative strategy has shown promise to repair cartilage, an avascular tissue in which cells experience hypoxia. Hypoxia is known to improve the early chondrogenic differentiation of stem cells. Therefore, this study aimed to determine whether hypoxia preconditioning could be used to enhance the regenerative potential of the combination of buccal fat pad stem cells (BFPSCs) and bilayer chitosan-based hydrogel scaffold for articular cartilage repair., Materials and Methods: Human BFPSCs were seeded on the bilayer chitosan-based hydrogel scaffolds in the culture medium. The viability and proliferation of cells on the scaffolds were monitored using scanning electron microscopy (SEM), MTT assay, and DAPI staining. Hypoxia preconditioned BFPSCs-seeded scaffolds were transplanted into rabbit articular cartilage knee defects for 12 weeks. The newly formed tissue was evaluated by cartilage-specific immunohistological analysis and histological staining., Key Findings: It was found that the chondrogenic differentiation and osteochondral conjunction in articular cartilage defect via BFPSCs-seeded bilayer scaffolds was enhanced by hypoxic preconditioning compared to a normoxic environment., Significance: Based on our study, the integrity with subchondral bone in osteochondral defect was enhanced by BFPSCs on bilayer scaffold. Thus, this study provides evidence on the design of preconditioned cell-seeded bilayer hydrogels for articular cartilage regeneration., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Mesenchymal stem cells (MSCs) on injectable hydrogels are mostly used to regenerate articular cartilage, which would have a variety of outcomes. Chondrocyte extracellular vesicles (EVs) have attracted many attentions for their chondrogenic differentiation capacity; however, the roles of EVs in both chondrogenic differentiation of MSCs and cartilage regeneration are poorly understood yet. In the current study, to investigate the differentiation effects of human articular chondrocyte EVs on adipose-derived MSCs, they were cultured in injectable chitosan-hyaluronic acid (CS-HA) hydrogel and then treated with chondrocyte EVs for 21 days. The continuous treatment of EVs performed on MSCs increased chondrogenic genes' expressions of SOX9 and COL2A1 and induced expression of Col II protein. In addition, glycosaminoglycans secretion was detected in the EV-treated MSCs after about 14 days. The therapeutic efficiency of this hydrogel and EVs was studied in a rabbit osteochondral defect model. MRI results revealed that the cartilage regeneration capacity of EV-treated MSCs with CS-HA hydrogel was greater than the untreated MSCs or the EV-treated MSCs without hydrogel. Moreover, histological results showed hyaline-like cartilage in the CS-HA/MSC and CS-HA/EV/MSC groups in the cartilage defect sites. These findings suggested that the chondrocyte-EVs and CS-HA hydrogel could provide the preferable niche for chondrogenic differentiation of MSCs and cartilage regeneration in osteoarthritis cartilage injuries., (© 2021 IOP Publishing Ltd.)

Electrospun composite scaffolds show high ability to be used in regenerative medicine and drug delivery, due to the nanofibrous structure and high surface area to volume ratio. In this study, we used nanofibrous scaffolds fabricated by chitosan (CS), poly(vinyl alcohol) (PVA), carbopol, and polycaprolactone using a dual electrospinning technique while curcumin (Cur) incorporated inside of the CS/PVA fibers. Scaffolds were fully characterized via scanning electron microscopy, water contact angle, tensile measurement, hydration, protein adsorption, and wrinkled tests. Furthermore, viability of the buccal fat pad-derived mesenchymal stem cells (BFP-MSCs) was also investigated using MTT assay for up to 14 days while cultured on these scaffolds. Cell cycle assay was also performed to more detailed evaluation of the stem cells growth when grown on scaffolds (with and without Cur) compared with the culture plate. Results demonstrated that Cur loaded nanofibrous scaffold had more suitable capability for water absorption and mechanical properties compared with the scaffold without Cur and it could also support the stem cells viability and proliferation. Cur release profile showed a decreasing effect on BFP-MSCs viability in the initial stage, but it showed a positive effect on stem cell viability in a long-term manner. In general, the results indicated that this nanofibrous scaffold has great potential as a delivery of the Cur and BFP-MSCs simultaneously, and so holds the promising potential for use in various regenerative medicine applications., (© 2019 Wiley Periodicals, Inc.)

In this study, a unique stimuli-responsive hydrogel nanocomposite was prepared via surface reversible addition fragmentation chain transfer (RAFT) copolymerization of acrylic acid and N‑isopropyl acrylamide onto chitosan and subsequent in situ synthesis of magnetic Fe 3 O 4 nanoparticles. In the next age, the structure and morphology of synthetic magnetic nanocomposite were characterized by FTIR, XRD, VSM, SEM, and TEM techniques. The modified hydrogel nanocomposite was employed as an excellent carrier for controlled releasing of anticancer drug doxorubicin (DOX). The results indicated that the maximum of DOX loading efficiency of nanocomposite was 89%. Notably, in vitro drug release studies showed that DOX was released in a sustained-release manner for the nanocomposites. From the results of in vitro release studies, dual temperature and pH responsiveness of the nanocomposite was demonstrated, and 82% of total DOX was released from the hydrogel within 2 days. Due to the unique structures and properties, the chitosan-based nanocomposite may be utilized as a promising drug carrier for controlled and sustained release of anticancer drugs., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The current study was aimed to investigate the combined effects of chitosan (Ch) and pomegranate peel extract (PPE) on the overall quality of rainbow trout (Oncorhynchus mykiss) during the frozen storage. Fresh gutted fishes were dipped for 60s either in sterile distilled water (Control) or in chitosan combined with 0 (Ch), 1% (Ch+PPE1), 2% (Ch+PPE2), or 4% (Ch+PPE4) (w/v) PPE. Microbial, chemical, textural and sensory properties of the samples were intermittently checked during six months. At the end of storage, the highest and lowest values of psychrophilic, lactic acid bacteria and mold counts in the control and Ch+PPE4 groups were respectively, 5.14±0.19, 2.91±0.00; 3.11±0.03, 0.33±0.57 and 2.86±0.19, 2.24±0.13. Our results showed the best performance to prevent the oxidation of fats and protein was in the chitosan combined with 4% PPE, however, due to its undesirable color, the chitosan combined with 2% PPE was preferred. A considerable improvement in the chemical, microbial, sensory and textural properties was shown in the Ch+PPE2 groups. Therefore, this active coverage was suggested as safe to store rainbow trout up to 6 months frozen storage., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Migration of fibroblasts into wound area is a critical phenomenon in wound healing process. We used an appropriate system to fabricate an electrospun bioactive scaffold with controlled release of PDGF-BB in order to induce migration of primary skin fibroblast cells. First of all, protein-loaded chitosan nanoparticles based on ionic gelation interaction between chitosan and sodium tripolyphosphate were prepared. Then polycaprolactone electrospun fibers containing chitosan nanoparticles or PDGF-BB-loaded chitosan nanoparticles were prepared. Cellular attachment and morphology of cells seeded on scaffolds with or without PDGF-BB were evaluated by using a fluorescence microscope and scanning electron microscopy. Cells were well-oriented 72 h after seeding on the scaffolds containing PDGF-BB. The mean aspect ratio of populations on scaffold containing PDGF-BB-loaded chitosan nanoparticles was significantly greater than those on the scaffold containing chitosan nanoparticles but no PDGF-BB. Furthermore, the Arp2 gene, which is involved in cell protrusion formation, showed about three times more expression at mRNA level, in cells seeding on PDGF-BB-containing scaffold compared to cells seeding on scaffold containing only chitosan nanoparticles, using Real Time PCR test. Finally, under agarose migration assay results demonstrated that cells' chemotaxic behavior was more toward scaffold containing PDGF-BB compared to the PDGF-BB alone or FBS group. In addition, in terms of distance, the cell mass could grow faster, in response to scaffold containing PDGF-BB compared to FBS or PDGF-BB alone; however, the number of migrating cells might be the same or significantly higher in the latter groups.

The effects of essential oil (EO) of oregano and chitosan on the microbial quality and growth inhibition of Listeria monocytogenes and Escherichia coli O157:H7 in the Iranian traditional ready-to-barbecue chicken was evaluated. Thus, three groups of samples were prepared. One of them was considered to evaluate for aerobic plate count (APC), lactic acid bacteria (LAB), psychrophilic and enterobacteriacae counts and the other two groups were inoculated with E. coli O157:H7 and L. monocytogenes 4b to investigate the effect of oregano EO and chitosan on pathogenic bacteria. All groups were stored at 3, 8 and 20°C. Oregano showed antibacterial effects against APC, LAB, psychrophilics, enterobacteriacae and E. coli O157:H7, whereas, such an effect was not observed against L. monocytogenes. Chitosan individually did not show an inhibitory effect on the spoilage-inducing bacteria and E. coli, but was effective against L. monocytogenes. Using chitosan and oregano EO in combination can reduce the number of spoilage and safety indicators and also the two food-borne pathogens in ready-to-barbecue chicken meat., (Copyright © 2015 Elsevier B.V. All rights reserved.)