Human bone marrow mesenchymal stem cell behaviors on PCL/Gelatin nanofibrous scaffolds modified with a collagen IV-derived RGD-containing peptide

We introduce an RGD [Arg-Gly-Asp]-containing peptide of collagen 4 origin that possesses potent cell adhesion and proliferation properties. In this experimental study, the peptide was immobilized on an electrospun nanofibrous polycaprolactone/gelatin [PCL/Gel] hybrid scaffold by a chemical bonding approach to improve cell adhesion properties of the scaffold. An iodine- modified phenylalanine was introduced in the peptide to track the immobilization process. Native and modified scaffolds were characterized with scanning electron microscopy [SEM] and fourier transform infrared spectroscopy [FTIR]. We studied the osteogenic and adipogenic differentiation potential of human bone marrow-derived mesenchymal stem cells [hBMSCs]. In addition, cell adhesion and proliferation behaviors of hBMSCs on native and peptide modified scaffolds were evaluated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide [MTT] assay and 4',6-diamidino-2-phenylindole [DAPI] staining, and the results compared with tissue culture plate, as the control. FTIR results showed that the peptide successfully immobilized on the scaffold. MTT assay and DAPI staining results indicated that peptide immobilization had a dramatic effect on cell adhesion and proliferation. This peptide modified nanofibrous scaffold can be a promising biomaterial for tissue engineering and regenerative medicine with the use of hBMSCs

[Study of hBMSC adhesion and proliferation on RGD-modified polycaprolactone/gelatin nanofibrous scaffold]

In this study we introduced an RGD-containing peptide of collagen IV origin that possesses potent cell adhesion and proliferation properties. This peptide was immobilized on a nanofibrous polycaprolactone/gelatin scaffold after which we analyzed human bone marrow-derived mesenchymal stem cells [hBMSCs] adhesion and proliferation on this peptide-modified scaffold. Nanofibrous scaffold was prepared by electrospinning. The peptide was synthesized by solid-phase peptide synthesis and immobilized on electrospun nanofibrous a polycaprolactone/gelatin scaffold by chemical bonding. Native and modified scaffolds were characterized with Scanning Electron Microscope [SEM] and Fourier-Transform Infra-red Spectroscopy [FTIR]. Adhesion and proliferation of hBMSCs on native and modified scaffolds were analyzed by the Methylthiazol Tetrazolium [MTT] assay. SEM images showed that electrospun scaffolds had homogenous morphology and were 312 +/- 89 nm in diameter. There was no significant difference in scaffold morphology before and after peptide immobilization. FTIR results showed that the peptide was successfully immobilized on the scaffold. Based on MTT assay, cell adhesion studies indicated that peptide immobilization improved cell adhesion on RGD-modified scaffolds at all corresponding time points [p<0.05]. RGD immobilization led to increased cell proliferation potential of the scaffold compared with tissue culture plate and native scaffold [P<0.05]. This novel peptide and modified nanofibrous scaffold, having improved cell adhesion and proliferation properties, can be used for tissue engineering and regenerative medicine by using hBMSCs.