Effect of Electrospun Collagen/Chitosan Composite Nanofibrous Membrane on Osteogenesis for Bone Regeneration
Li Xiaojing1,2 ,Gao Bo1, Dong Yan1* ,Gou Zhongru3, Miao Yuwen1
1Department of Prosthetic Dentistey, Second Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310009, China; 2Department of Stomatology, Children′s Hospital, College of Medicine, Zhejiang University, Hangzhou 310006, China 3Zhejiang-California International Nanosystems Institute, Zhejiang University, Hangzhou 310029, China
Abstract：The aims of this study are to prepare collagen/chitosan composite nanofibrous membranes, and to examine their biocompatibility and effect of osteogenic differentiation for bone marrow mesenchymal stem cells(BMSCs). Take acetic acid as solvent,poly(ethylene oxide) (PEO) as plasticizer, the collagen nanofibrous membranes (the mass rate of collagen/PEO, 4∶1) and various collagen/chitosan nanofibrous membranes (the mass rate of collagen/chitosan/PEO, 5∶1∶4,5∶2∶3,5∶4∶1) were fabricated through electrospinning technique. The surface morphology of membranes were observed by scanning electron microscopy (SEM). BMSCs were cultured on the collagen nanofibrous membranes and the optimized collagen/chitosan nanofibrous membranes, the cell proliferation and osteogenic makers including alkaline phosphate(ALP), collagen content, osteocalcin (OCN) and mineralized matrix deposits were detected respectively. SEM showed collagen nanofibers and collagen/chitosan nanofibers of mass ratio of 5∶1∶4 had a mean fiber diameter. Both collagen nanofibrous membranes and collagen/chitosan nanofibrous membranes supported adhesion, proliferation and osteogenic differentiation of the BMSCs. The collagen/chitosan nanofibrous membranes induced higher proliferation of BMSCs than collagen nanofibrous membranes. The collagen contents were higher in cells cultured on the collagen/chitosan nanofibrous membranes for 14 days as compared to collagen nanofibrous membranes (p<0.05). Similarly, cells cultured on collagen/chitosan nanofibrous membranes had the higher expression of ALP, OCN and mineralized matrix deposits than that on collagen nanofibrous membranes. Our results demonstrated that the collagen/chitosan composite nanofibrous membranes promoted adhesion, proliferation and osteogenic differentiation of BMSCs,implying their promising applications for bone regeneration.
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