In silico analysis, preparation and evaluation of exosomes for scaffold-based tissue repair

Abstract

Exosomes derived from mesenchymal stem cells (MSCs) are highly researched therapeutic cell-free vesicles because of their ability to induce tissue repair by delivering bioactive molecules such as proteins, lipids, messenger RNAs and microRNAs. Their minimal immune rejection, increased stability, and lower risk of tumorigenesis provide an edge over the conventional stem cell therapy for the treatment of musculoskeletal injuries. In the present work we blended computational thinking with experimental work, to explore molecular targets linked to exosome biogenesis and tissue regeneration. We also aimed to prepare and characterize MSC-derived exosomes, and to design a bioactive corneal scaffold. More specifically, bioinformatics analyses were done, including differential gene expression, functional enrichment, protein–protein interaction network analysis, and molecular docking, in order to spot key molecular targets that relate to osteogenesis, myogenesis, and exosome biogenesis. In this work, bone marrow derived mesenchymal stem cells (BM-MSCs) were isolated from BALB/c mice and grown in standard laboratory conditions. Further, exosomes were isolated from the conditioned medium and characterized with respect to particle size, concentration, and expression of exosomal markers. For scaffold fabrication, polyvinyl alcohol–polyvinyl pyrrolidone (PVA–PVP) and polyvinyl alcohol–polycaprolactone (PVA–PCL), were incorporated with retinoic acid, raloxifene, dimethyl sulfoxide (DMSO), and graphite, to help enhance both their physico-chemical features, and biological responses. The scaffolds were evaluated for transparency, mechanical strength, swelling, degradation, contact angle, suturability, antibacterial activity, and biocompatibility using human corneal epithelial (HCE) cells. The current research uses bioinformatics tools along with experimental studies to advance the knowledge about tissue regeneration, regenerative medicine applications and enhances our knowledge of exosome-mediated tissue healing.