Design and Fabrication of Demineralized and Decellularized Hydrogels Loaded with Titanium Dioxide Nanoparticles and Curcumin for Osteoblastic Differentiation of Adipose-Derived Mesenchymal Stem Cells

Abstract

Abstract Background: Bone tissue engineering (BTE) is a prospective method for providing effective scaffolds for the treatment of bone injuries. A novel hydrogel-based composite can be used as functional biomimetic and biodegradable scaffold that amends osteoblastic differentiation of adipose-derived mesenchymal stem cells (ADMSCs). Nanostructured materials such as titanium dioxide nanoparticles (nTiO2) have been used as bone substitute materials because of their remarkable physicochemical features such as biological compatibility, great stability, and low toxicity. Curcumin has been shown to regulate osteogenic activities. However, curcumin could regulate influence osteogenesis remain to be explained. Methods: In this study, titanium dioxide nanoparticles (TiO2) and curcumin (Cur) coembedded extracellular matrix (ECM) hydrogel to fabricate Hy/Ti/Cur composite. In this way, the fresh bovine femur was demineralized and decellularized, then by digestion of these matrices, ECM hydrogel was obtained. The fabricated hydrogels were characterized to investigate morphological features and chemical configuration. In vitro cytotoxicity and osteogenic differentiation of ADMSCs on these composite were also evaluated. The experiments were performed in triplicate. Analyses of the outcomes were evaluated in GraphPad Prism Software (version 9.0.0 121). Results: Our results showed that the hydrogels were cytocompatible, and the cell viability on the hydrogels was elevated compared to the control. The synergistic effect of TiO2 and Cur co-embedded on ECM hydrogel (Hy/Ti/Cur) stimulates bone differentiation markers, such as Runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), type I collagen (Col), Bone morphogenetic protein 2 (BMP2) and Osterix (OSX) in ADMSCs cultured in normal and osteogenic medium. Moreover, Alkaline Phosphatase (ALP) activity and calcium deposition of ADMSCs cultured on engineered hydrogels were increased, where it is recommended as an attractive scaffold in bone tissue engineering. Titanium as an effective combination in the hydrogel structure increased scaffold’ mechanical property. Conclusions: SEM micrographs of Hy/Ti/Cur revealed that the Cur and titanium were distributed and loaded into the hydrogel networks, representing a mesh-like network structure that mimics the native architecture of bECM. Therefore, it can be expected the huge cell proliferation inside the materials.