Extracellular Matrix Based 3D Scaffold for Directing the Fate of Adult Stem Cells into Osteogenic Lineage

Abstract

Background Bone defect causes softening of bone, reduction of bone density and mass, and degenerating bone microstructure which directly leads to bone fractures. Tissue-derived engineered cell-seeded hydrogel scaffolds are considered a promising strategy for the treatment of such defects. Among the most important objectives is to develop a unique biological tissue derived scaffolding material that is easily manipulated, efficient for cell attachment, and biocompatible, as well as supports cell growth, proliferation, and differentiation along osteogenic lineage. Methods This research describes a technique for the preparation of decellularized hydrogels. The detergent-based procedure using Triton-X-100 reported here effectively eliminates cellular proteins and antigens, as well as nucleic acid, while causing minimal damage to the extracellular matrix (ECM). The final product contains natural ECM components that guide stem cell differentiation by simulating the natural tissue microenvironment. Umbilical cord-derived mesenchymal stem cells (MSCs) were seeded in the hydrogel. The cell-hydrogel constructs were cultured in basal or osteogenic media for 28 days. Thereafter, mRNA and protein expression of osteogenic markers were analyzed. Results The results indicated that the scaffold had a fibrous structure with a relatively homogenous exterior. The cells seeded hydrogel after 28 days of culture in basal media as well as in osteogenic inductive media displayed higher mRNA and protein expression of osteogenic markers. Moreover, calcium deposits were also observed which further confirmed osteogenic differentiation. Conclusion The present findings demonstrate that the hydrogel is biocompatible with MSCs and possesses osteo-conductive and osteo-inductive capability in vitro. It is potentially useful for bone tissue engineering.