Apigenin Release from Chitosan/Gelatin Membranes Promotes Osteogenic Differentiation of Adipose-Derived Mesenchymal Stem Cells

Abstract

Purpose

Apigenin is a widespread phytochemical with beneficial effects on osteoblastic differentiation. However, short half-life and unstable chemical structure restrict apigenin application in bone tissue engineering applications. Here, we investigated the impact of apigenin-loaded chitosan/gelatin (Api. Cs/Gel) membranes on the osteogenic differentiation of human adipose-derived mesenchymal stem cells (hADMSCs).

Methods

Api10. Cs/Gel and Api25. Cs/Gel membranes were fabricated using the solution casting method, followed by characterizing their physicochemical and biological properties. hADMSCs were isolated from healthy donors and characterized using flow cytometry. hADMSCs were seeded onto Api. Cs/Gel membranes and cultured under osteogenic differentiation for 7 and 21 days. The expression of osteogenic markers ALP, RUNX2, OCN, and COL1 was assessed using real-time PCR, and calcium mineralization was analyzed using the quantitative Alizarin red S staining.

Results

The Api. Cs/Gel membranes were successfully fabricated, and characterization data confirmed their structural uniformity, chemical homogeneity, cross-linking, and apigenin incorporation. Membranes exhibited favorable degradation, swelling ratio, and long-term apigenin release. The membranes were non-toxic and supported hADMSCs attachment, viability, and proliferation. The expression of ALP, RUNX2, OCN, and COL1 and cellular mineralization increased on day 21, and significant overexpression was observed in hADMSCs seeded onto Apigenin-loaded membranes.

Conclusion

Cs/Gel membranes provided an approving matrix for cellular interactions and apigenin inclusion, followed by a sustained release associated with enhanced osteogenic differentiation of hADMSCs and matrix mineralization.