Calcium-Silicate-Incorporated Gellan-Chitosan Induced Osteogenic Differentiation in Mesenchymal Stromal Cells

Krishnamurithy Genasan; Mohammad Mehrali; Tarini Veerappan; Sepehr Talebian; Murali Malliga Raman; Simmrat Singh; Sasikumar Swamiappan; Mehdi Mehrali; Tunku Kamarul; Hanumantha Rao Balaji Raghavendran

doi:10.3390/polym13193211

Calcium-Silicate-Incorporated Gellan-Chitosan Induced Osteogenic Differentiation in Mesenchymal Stromal Cells

Polymers (Basel). 2021 Sep 22;13(19):3211. doi: 10.3390/polym13193211.

Authors

Affiliations

¹ National Orthopaedic Centre of Excellence in Research and Learning (NOCERAL), Tissue Engineering Group (TEG), Department of Orthopaedic Surgery, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia.
² Faculty of Engineering Technology, Department of Thermal and Fluid Engineering (TFE), University of Twente, 7500 AE Enschede, The Netherlands.
³ Faculty of Engineering, School of Chemical and Biomolecular Engineering, The University of Sydney, Sydney, NSW 2006, Australia.
⁴ Nano Institute (Sydney Nano), The University of Sydney, Sydney, NSW 2006, Australia.
⁵ Materials Chemistry Division, School of Advanced Sciences, VIT University, Vellore 632014, Tamil Nadu, India.
⁶ Department of Mechanical Engineering, Technical University of Denmark, 2800 Kgs Lyngby, Denmark.
⁷ Advanced Medical and Dental Institute (AMDI), University Sains Malaysia, Bertam, Kepala Batas 13200, Penang, Malaysia.
⁸ Faculty of Clinical Research, Central Research Facility, Sri Ramachandra Institute of Higher Education and Research Porur, Chennai 600116, Tamil Nadu, India.

Abstract

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.

Keywords: bone; bone tissue engineering; calcium silicate; chitosan; gellan; osteoinduction.

Grants and funding

FRGS-FP016, TR001-2016/This study was funded by FRGS-FP016, Fundamental Research Grant Scheme (FRGS), and Trans-Disciplinary Research Grant Scheme (TR001-2016).