Enhanced osteogenic differentiation of human bone marrow-derived mesenchymal stem cell on polycaprolactone/ brushite nanofibrous scaffold
Enhanced osteogenic differentiation of human bone marrow-derived mesenchymal stem cell on polycaprolactone/ brushite nanofibrous scaffold
Seyedeh Sara Shafiei,1,*yeganeh Nikakhtar,2Mehrnoush Fathi-roudsari,3
1. National institute of genetic engineering and biotechnology 2. National institute of genetic engineering and biotechnology 3. National institute of genetic engineering and biotechnology
Introduction: Bone tissue engineering (BTE) aims to develop effective methods for repairing or replacing damaged bone tissue based on using three factors including biological molecules, cells, and biocompatible scaffolds, simultaneously.
Methods: In this study, composite scaffolds consisting of dicalcium phosphate dihydrate (DCPD, brushite) as a bone phase mineral precursor with different weight percentages (0%, 1%, 3%, 5%, and 10%) in combination with polycaprolactone (PCL) were fabricated by electrospinning technique. To assess the properties of the fabricated electrospun scaffolds, different techniques were employed. The morphology and mechanical behavior of scaffolds were characterized using scanning electron microscopy (SEM) and tensile strength test, respectively. The bioactivity of scaffolds was assessed in simulated body fluid (SBF). Adhesion, viability, proliferation, and differentiation of mesenchymal stem cells derived from the human bone marrow (hMSC) on scaffolds were investigated using electron microscopy (SEM), MTT assay, live-dead assay, alizarin red staining, alkaline phosphatase activity and, gene expression analysis by real-time PCR.
Results: The results showed that PCL/DCPD (3 Wt %) had the highest tensile strength (15.35 MPa) which indicates a significant increase compared to the pure PCL. Furthermore, hMSC seeded on scaffolds showed over 80% viability after 1, 3, 7 days of incubation. Also, the results showed that the addition of DCPD to the PCL significantly increased the alkaline phosphatase activity. The osteocalcin (OCN) gene expression in the composite scaffold showed a 6.1-fold increase compared to the pure PCL.
Conclusion: It is concluded that electrospun PCL/DCPD scaffolds with optimum concentration can be a proper candidate for bone tissue engineering applications.