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

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

doi:10.3390/polym13193211

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.

Highlights

As lifespan increases, injuries related to bones are common among elderly individuals, compromising their activities of daily living (ADLs) and posing lifestyle and economic challenges [1]
The attachment of human bone-marrow mesenchymal stromal cells (hBMSCs) seeded in GC-0 calcium silicate (CS), GC-10 CS, GC20 CS, and GC-40 CS scaffolds was evaluated using Scanning electron microscopy (SEM) imaging
Cell attachment was evidenced in all composite scaffolds (Figure 1b,d,f,h), and the cells demonstrated fibroblast-like appearance, as indicated by the red arrow

Summary

Introduction

Injuries related to bones are common among elderly individuals, compromising their activities of daily living (ADLs) and posing lifestyle and economic challenges [1]. Despite the fact that advancements have been achieved in bone tissue engineering to date, there are drawbacks to the current treatments [2]. This has led to the development of the biotechnological sector, with the aim of establishing biomaterials for bone tissue engineering. A considerable amount of attention has been paid to the use of biodegradable polymers for bone tissue engineering. These polymers were proven to induce the osteogenic differentiation in bone marrow derived cells [5]. Much focus has been given to polymers derived from natural sources owing to its chemical versatility and extracellular matrix that support excellent cellular interactions [6]

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Discussion

Conclusion