The osteogenic differentiation of Wharton’s jelly derived mesenchymal stem cell seeded on chitosan/hyaluronic acid multilayers to treat Osteogenic Imperfecta (OI)

Abstract

Osteogenesis Imperfecta (OI), a rare genetic condition characterized by brittle bones and an increased risk of fractures, currently has limited treatment options, primarily focusing on symptom management rather than providing a cure. Recognizing this challenge, bone tissue engineering has gained prominence as a potential solution. In this study, our aim is to investigate skeletal diseases, with a specific focus on Osteogenesis Imperfecta (OI), utilizing the bone tissue engineering triad. We adopted a comprehensive approach for bone regeneration in Osteogenesis Imperfecta (OI). This approach entailed the use of a combination of Chitosan-Hyaluronic Acid (CHI-HA) scaffolds and growth factors, including Dexamethasone, Beta-Glycerophosphate, and L-Ascorbic Acid, in conjunction with Wharton's jelly extraction. Our methodology included confirming the origin of mesenchymal cells and initiating osteogenic induction while constructing a CHI-HA scaffold. Subsequent analyses, including Alizarin Red staining, Real-time Polymerase Chain Reaction (RT-PCR), and Flow Cytometry, enabled us to assess osteogenic marker expression and mineralization. To assess mineralization, we conducted Alizarin Red staining, which provided confirmation of the presence of calcium deposits in our induced WJ-MSCs. Additionally, the RT-PCR results revealed a clear shift towards osteogenic differentiation, confirming the successful commitment to the osteogenic lineage, as indicated by a substantial increase in Alkaline Phosphatase (ALP) expression (19.47). Furthermore, it was noteworthy that we observed the expression of CD34 markers in osteoblasts in 19% of the cells on induced WJ-MSCs on CHI/HA scaffolds. The outcomes of this study indicate that the chitosan-hyaluronic acid (CHI-HA) blend, as natural polymers for scaffolds, shows promise in promoting osteogenic differentiation.