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Background To address the limitations of Cultivated Limbal Epithelial Transplantation (CLET) and the use of amniotic membrane (AM) in treating Limbal Stem Cell Deficiency (LSCD), we aimed to develop a Collagen/Silk Fibroin (Co/SF) scaffold enriched with Platelet-Rich Growth Factor (PRGF) to support the proliferation, maintenance, and differentiation of Wharton's jelly-derived mesenchymal stem cells (WJMSCs) into corneal epithelial cells (CECs). Method Scaffolds loaded with PRGF were evaluated through release studies, cytotoxicity assays, and cell differentiation. The proliferation and differentiation of WJMSCs and Limbal Epithelial Stem Cells (LESCs) were investigated using MTT assays, real-time PCR and immunostaining. Results The PRGF-loaded Co/SF scaffold significantly promoted the proliferation of both WJMSCs and LESCs in a concentration-dependent manner. Real-time PCR and immune staining revealed a significant increase in the expression of P63, ABCG2, and cytokeratin 3/12 markers in WJMSCs, a significant decrease in the expression of P63 and ABCG2, and a significant increase in the expression of cytokeratin 3/12 markers indicating successful differentiation into CECs. Conclusion The WJMSC cultured on PRGF-enriched Co/SF scaffold demonstrates potential as a viable alternative to conventional CLET, offering a promising strategy for corneal tissue regeneration. [ABSTRACT FROM AUTHOR]

Rapid wound healing using platelet‐rich growth factors (PRGF), stem cells, and probiotics is important due to their positive effects on skin regeneration and antibacterial activities. A novel method was employed to fabricate a biomaterial multi‐layered electrospun scaffold composed of polyurethane, PRGF, and gelatin fibers. This scaffold was used to induce mesenchymal stem cells with fibroblast cells (co‐cultured cells) and Lactobacillus plantarum. The scaffolds were analyzed for physical, mechanical, and biological characteristics. The results showed that the nanocomposite scaffolds had a higher Young's modulus and decreased contact angle. Scanning electron microscope and MTT assays demonstrated improved spreading, density, and proliferation of co‐culture cells and L. plantarum on the PU/PRGF/gelatin/PU scaffolds. Additionally, L. plantarum showed antibacterial effects on Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli. Furthermore, in vivo studies showed that the PU/PRGF/gelatin/PU scaffold with Co and L.p was implanted on the full‐thickness skin wound of the rat's dorsal area, resulting in high pathological scores and aesthetic macroscopic results that demonstrated greater wound repair efficacy compared to the control groups. Based on the results, PU/PRGF/gelatin/PU containing the co‐culture cells and L. plantarum showed better potential for cell attachment, angiogenesis, proliferation, prevention of infection, and accelerated wound healing in rats. Highlights: Multi‐layered scaffold designed to mimic the architectures found in tissue layers.Co‐culture involves diverse cell populations with varying levels of interaction.Antibacterial activity inhibits target bacteria growth without harming nearby tissues.Full‐thickness wounds penetrate beyond the dermis and epidermis layers of the skin.Wound healing involves a number of stage‐specific molecular pathways. [ABSTRACT FROM AUTHOR]

Purpose: A hemocompatible substrate can offer a wonderful facility for nitric oxide (NO) production by vascular endothelial cells in reaction to the inflammation following injuries. NO inhibits platelet aggregation this is especially critical in small-diameter vessels. Methods: The substrate films were made of polyurethane (PU) in a casting process and after plasma treatments, their surface was chemically decorated with polyethylene glycol (PEG) 2000, gelatin, gelatin-aspirin, gelatin-heparin and gelatin-aspirin-heparin. The concentrations of these ingredients were optimized in order to achieve the biocompatible values and the resulting modifications were characterized by water contact angle and Fourier transform infra-red (FTIR) assays. The values of NO production and platelet adhesion were then examined. Results: The water contact angle of the modified surface was reduced to 26 ± 4⸰ and the newly developed hydrophilic chemical groups were confirmed by FTIR. The respective concentrations of 0.05 mg/ml and 100 mg/mL were found to be the IC50 values for aspirin and heparin. However, after the surface modification with aspirin, the bioactivity of the substrate increased in compared to the other experimental groups. In addition, there was a synergistic effect between these reagents for NO synthesis. While, heparin inhibited platelet adhesion more than aspirin. Conclusion: Because of the highly hydrophilic nature of heparin, this reagent was hydrolyzed faster than aspirin and therefore its influence on platelet aggregation and cell growth was greater. Taken together, the results give the biocompatible concentrations of both biomolecules that are required for endothelial cell proliferation, NO synthesis and platelet adhesion. [ABSTRACT FROM AUTHOR]

Introduction: Migration of fibroblast cells in wound areas is a critical aspect of the wound healing process. Employment of enhanced green fluorescent protein (EGFP) labeled fibroblast cells facilitates real-time monitoring and functional evaluation of these cells in both in vitro and in vivo settings. Plasma rich in growth factor (PRGF) is a potent accelerator of wound healing; therefore, in this study, a novel method to fabricate an electrospun bioactive scaffold containing PRGF was employed to induce in vitro cell proliferation and migration. Methods: First, the EGFP reporter gene was integrated into the AAVS1 locus of fibroblast cells using CRISPR/Cas9 system. Then, PRGF was obtained from platelet-rich plasma, and a multi-layered scaffold was fabricated using polyurethane-cellulose acetate (PU-CA) fibers as the outer layers and PRGF-containing gelatin fibers were located in the internal layer like a central strip. Scanning electron microscopy (SEM), tensile, water contact angle, and FTIR tests were performed to assess the characteristics of the scaffolds. The EGFP targeted cells were cultured on scaffolds with or without PRGF to investigate their viability, toxicity, and migration pattern in response to the release profile. Results: Fluorescence images showed that the number of migrating cells on scaffold containing PRGF was more significant than PU-CA scaffold up to day 6. Increased expression of SGPL1 , DDR2 , and VEGF genes was also observed on the scaffold containing PRGF compared to PU-CA using real-time polymerase chain reaction (PCR) analysis with around 3-, 2-, and 2-fold enhancement, respectively. Conclusion: The current scaffold provides the appropriate template for cell attachment and migration. In addition, the present results highlight the potential of reporter gene targeting for the in vitro analysis of biological processes such as migration. [ABSTRACT FROM AUTHOR]

Background: Regenerative endodontics requires an innovative delivery system to release antibiotics/growth factors in a sequential trend. This study focuses on developing/characterizing a thermoresponsive core-shell hydrogel designed for targeted drug delivery in endodontics., Methods: The core-shell chitosan-alginate microparticles were prepared by electrospraying to deliver bone morphogenic protein-2 for 14 days and transforming growth factor-beta 1 (TGF-β1) for 7-14 days. Methylcellulose (MC) and gelatin were utilized to create the core-shell hydrogel to load a modified triple antibiotic combination (penicillin G/metronidazole/ciprofloxacin (PMC)) and growth factor-loaded microparticles in the shell and the core compartments, respectively. Morphological assessment, core-shell structural analysis, FTIR analysis, rheological analysis, swelling, and degradation rate studies were conducted for characterization. The viability of dental pulp stem cells (DPSCs) upon antibiotic exposure, antibacterial activity, and release studies of PMC and growth factors were investigated. Cellular studies (cell viability, alkaline phosphatase (ALP) activity, osteo/odontoblast gene expression (using Reverse transcription-polymerase chain reaction (RT-PCR)) and in vivo studies (inflammatory response and differentiation potential of the developed hydrogel by subcutaneous implantation in rats via histological examination) were assessed., Results: The hydrogel showed a porous microstructure with interconnected pores. Core-shell structure analysis confirmed the successful extrusion of the MC hydrogel to the surface. FTIR analysis revealed interactions between MC and gelatin. Rheological analysis indicated time-dependent gel formation, supporting thermosensitivity at 37 °C. Swelling occurred rapidly, and degradation reached 62.42% on day 45. Further, antibiotics exhibited no cytotoxicity on DPSCs. Sequential release of antibiotics and growth factors was observed for up to 5 and 14 d, respectively. The hydrogel showed antibacterial activity. DPSCs exhibited increased proliferation, ALP activity, and odontoblast gene expression. In vivo studies showed that the biocompatible drug-loaded hydrogel exhibited more mineralization than the control., Conclusions: The developed core-shell hydrogel containing PMC and growth factor-loaded core-shell microparticles provided a versatile and biocompatible platform for sequential drug delivery in regenerative endodontics. The system demonstrates promising characteristics for dentin regeneration, making it a potential candidate for clinical applications., (© 2024 The Author(s). Published by IMR Press.)

Different types of stem cells have remarkable characteristics such as high proliferation rate, multi/pluripotency, self-renewal, and broad differentiation that can effectively treat diseases, cancers, and damage. Despite abundant therapeutic applications of stem cells in medical science, numerous risks threaten stem cell transplantation. Tumor development, immune response, cellular senescence, dosage effects, and administration timing are critical risks that should be considered in stem cell therapy. Hence, an investigation of possible risks is required before utilizing stem cell-based medicinal products in the clinical phase and human trials. This review aims to survey the literature and perspectives on the advantages and risks associated with pluripotent and multipotent stem cells., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)