Preparation and characterization of the ostrich eggshells membrane for osteogenic induction of human mesenchymal stromal cells

In spite of advanced innovative therapeutic strategies using mesenchymal stromal cells (MSCs) to improve bone regeneration in recent years, finding a feasible and efficient extracellular matrix (ECM) to boost the healing of bone fractures with the proper migration and osteogenic capability is still critical in regenerative medicine. Natural scaffolds supposed to provide an appropriate ۳D microenvironment for bone regeneration. Present study aimed to evaluate the potency of using ostrich eggshells as scaffold for the growing human mesenchymal stromal cells and induction of the osteogenic differentiation.Materials and Methods: The structural properties of the composites were characterized by Fourier transform infrared (FTIR), Field emission scanning electron microscopy (FESEM), Atomic force microscopy (AFM), Brunauer Emmett Teller (BET) after preparation steps, cell adhesion, cell proliferation,cytotoxicity, and biodegradation was assessed. as well as evaluate osteogenic differentiation adipose-derived mesenchymal stem seeded on a scaffold. were estimated by alizarin red staining, and alkaline phosphatase activity measurement Also, cells gene expression analysis such as COLIA۱, ALP, OCN, and RunX۲ was evaluated using real time PCR. Results:structural analysis of the scaffolds showed that ostrich eggshell membrane is capable of inducing bone differentiation in terms of surface roughness, mechanical properties, and interconnected pores as well as low toxicity. Proposed natural matrix exhibited a proper osteocyte differentiation of the MSCs determined with increased cell's ALP activity and mineralization. Significant increase in expression of the osteogenic markers, including Runx۲, ALP, collagen-۱ (Col-I), and osteocalcin genes in the cells cultured on ostrich eggshell membrane compare to that of cultured on the culture plate have demonstrated its immense bone regenerative potential. Conclusion: Data presented here reports using ostrich eggshell membrane as ECM is a safe and cost-effectivestrategy for the first time to optimize MSC-based bone regeneration.

Keywords:

Bone Regeneration , Ostrich Eggshell Membrane , Tissue Engineering , Mesenchymal Stromal Cells