Osteogenic Differentiation of MSCs on Fibronectin-Coated and nHA-Modified Scaffolds

Abstract

The increasing demand for biocompatible bone substitutes has made it a priority to tissue engineering and regenerative medicine scientists. Combination of minerals, growth factors, and extracellular matrix (ECM) proteins with nanofibrous scaffolds is a potential promising strategy for bone reconstruction and clinical applications. In this study, nanohydroxyapatite (nHA) was incorporated in electrospun nanofibrous polycaprolactone (PCL) scaffolds coated with fibronectin (Fn). The potential bone regeneration capacities of these scaffolds were evaluated in vitro and in vivo using mouse mesenchymal stem cells (mMSCs). The interconnected pores and proper mechanical characteristics of the fabricated electrospun PCL mats in combination with nHA and Fn provided suitable environment for cell attachment, proliferation, and enhanced osteogenic differentiation. The synergistic effect of Fn and nHA on the both in vitro and in vivo increase of calcium deposition was assessed by biochemical analysis. In addition, alkaline phosphatase (ALP) activity in nHA-incorporated PCL scaffold (PCL/nHA) and Fn-coated PCL/nHA (PCL/nHA/Fn) were significantly higher in comparison to the control group. Quantitative real-time reverse transcription polymerase chain reaction (RT-PCR) analyses of important bone-related genes (ALP, osteocalcin, osteopontin, and Runx2) revealed that Fn has additive effect on promoting the osteogenic differentiation. The aforementioned results indicated that nanofibrous PCL/nHA scaffold coated with Fn is a promising candidate for bone-tissue engineering applications.

The Osteogenic Differentiation Effect of the FN Type 10-Peptide Amphiphile on PCL Fiber

“…Particularly, 5 μg·mL −1 of FNIII10-PA increased protein adhesion 3-fold compared with that of FNIII10. In many studies, PCL scaffold coated with FN remarkably enhanced cell adhesion, proliferation, and differentiation [ 23 , 24 , 25 ]. Drevelle et al found that the PCL film functionalized with RGD peptide (Ac-CGGNGEPRGDTYRAY-NH 2 derived from the bone sialoprotein) and easily adsorbed the serum adhesive proteins FN and vitronectin (VN), leading to increased cell adhesion and spreading by activating focal adhesion kinase (FAK) signaling [ 26 ].…”

“…FN was shown to have a powerful effect on both cell adhesion and differentiation of stem cells. For instance, Mohamadyar-Toupkanlou et al reported that nanofibrous PCL/HA scaffold coated with FN provided a suitable environment for proliferation as well as attachment and differentiation in mouse MSCs (mMSCs) [ 23 ]. Shekaran et al found that ECM-coated PCL microcarriers promoted human early mesenchymal stem cell growth, leading to cell expansion [ 24 ].…”

Bone tissue engineering: Adult stem cells in combination with electrospun nanofibrous scaffolds

“…In two separate studies that conducted by our group, we demonstrated that PCL and pulsed electromagnetic field have a synergistic effect on osteogenic differentiation potential of MSCs and human induced pluripotent stem cells (iPSCs) (Ardeshirylajimi & Khojasteh, ; Arjmand, Ardeshirylajimi, Maghsoudi, & Azadian, ). In another study, we showed that nano‐HA and fibronectin (Fn) could increase osteoinductivity and osteoconductivity of PCL nanofibrous scaffold by culturing of MSCs and ectopic new bone formation (; Mohamadyar‐Toupkanlou et al, ; Mohamadyar‐Toupkanlou, Vasheghani‐Farahani, Bakhshandeh, Soleimani, & Ardeshirylajimi, ). Thompson et al developed the PCL performance by magnesium ferrite (Mg‐ferrite) based composite nanofibers was synthesized using an Electrospinning technique.…”

Recent Achievements in Bone and Skin Tissue Engineering in Iran

“…Considering the fact that stem cell research in Iran has grown quiet well, the research in the field of bone tissue engineering has grown as well using bone marrow mesenchymal , adipose , and Wharton's jelly derived stem cells for regeneration of bone.…”