Calcium-Silicate-incorporated Gellan-Chitosan induced osteogenic differentiation in mesenchymal stromal cells

• Main Authors: Genasan, Krishnamurithy, Mehrali, Mohammad, Veerappan, Tarini, Talebian, Sepehr, Malliga Raman, Murali, Singh, Simmrat, Swamiappan, Sasikumar, Mehrali, Mehdi, Kamarul, Tunku, Balaji Raghavendran, Hanumantha Rao
• Format: Article
• Published: MDPI 2021
• Subjects: QR Microbiology; R Medicine
• Online Access: http://eprints.um.edu.my/34343/
• Institution: Universiti Malaya

Gellan-chitosan (GC) incorporated with CS: 0% (GC-0 CS), 10% (GC-10 CS), 20% (GC-20 CS) or 40% (GC-40 CS) w/w was prepared using freeze-drying method to investigate its physicochemical, biocompatible, and osteoinductive properties in human bone-marrow mesenchymal stromal cells (hBMSCs). The composition of different groups was reflected in physicochemical analyses performed using BET, FTIR, and XRD. The SEM micrographs revealed excellent hBMSCs attachment in GC-40 CS. The Alamar Blue assay indicated an increased proliferation and viability of seeded hBMSCs in all groups on day 21 as compared with day 0. The hBMSCs seeded in GC-40 CS indicated osteogenic differentiation based on an amplified alkaline-phosphatase release on day 7 and 14 as compared with day 0. These cells supported bone mineralization on GC-40 CS based on Alizarin-Red assay on day 21 as compared with day 7 and increased their osteogenic gene expression (RUNX2, ALP, BGLAP, BMP, and Osteonectin) on day 21. The GC-40 CS-seeded hBMSCs initiated their osteogenic differentiation on day 7 as compared with counterparts based on an increased expression of type-1 collagen and BMP2 in immunocytochemistry analysis. In conclusion, the incorporation of 40% (w/w) calcium silicate in gellan-chitosan showed osteoinduction potential in hBMSCs, making it a potential biomaterial to treat critical bone defects.