title: Modulation of Bone-Specific Tissue Regeneration by Incorporating Bone Morphogenetic Protein and Controlling the Shell Thickness of Silk Fibroin/Chitosan/Nanohydroxyapatite Core-Shell Nanofibrous Membranes.
abstract: The presence of both osteoconductive and osteoinductive factors is important in promoting stem cell differentiation toward the osteogenic lineage. In this study, we prepared silk fibroin/chitosan/nanohydroxyapatite/bone morphogenetic protein-2 (SF/CS/nHAP/BMP-2, SCHB2) nanofibrous membranes (NFMs) by incorporating BMP-2 in the core and SF/CS/nHAP as the shell layer of a nanofiber with two different shell thicknesses (SCHB2-thick and SCHB-thin). The physicochemical properties of SCHB2 membranes were characterized and compared with those of SF/CS and SF/CS/nHAP NFMs. When tested in release studies, the release rate of BMP-2 and the concentration of BMP-2 in the release medium were higher for SCHB2-thin NFMs because of reduced shell thickness. The BMP-2 released from the nanofiber retained its osteoinductive activity toward human-bone-marrow-derived mesenchymal stem cells (hMSCs). Compared with SF/CS and SF/CS/nHAP NFMs, the incorporation of BMP-2-promoted osteogenic differentiation of hMSCs and the SCHB-thin NFM is the best scaffold during in vitro cell culture. Gene expression analysis by real-time quantitative polymerase chain reaction detected the evolution of both early and late marker genes of bone formation. The relative mRNA expression is in accordance with the effect of BMP-2 incorporation and shell thickness, while the same was reconfirmed through the quantification of bone marker protein osteocalcin. In vivo experiments were carried out by subcutaneously implanting hMSC-seeded SCHB2-thin NFMs and acellular controls on the back sides of nude mice. Immunohistochemical and histological staining confirmed ectopic bone formation and osteogenesis of hMSCs in SCHB2-thin NFMs. In conclusion, the SCHB2-thin NFM could be suggested as a promising scaffold for bone tissue engineering.
authors.affiliation: Department of Chemical and Materials Engineering, Chang Gung University , Kwei-San, Taoyuan 333, Taiwan Republic of China.
authors.forename: K T
authors.initials: KT
authors.lastname: Shalumon
authors.affiliation: Department of Chemical and Materials Engineering, Chang Gung University , Kwei-San, Taoyuan 333, Taiwan Republic of China.
authors.forename: Guo-Jyun
authors.initials: GJ
authors.lastname: Lai
authors.affiliation: Department of Chemical and Materials Engineering, Chang Gung University , Kwei-San, Taoyuan 333, Taiwan Republic of China.|Department of Plastic and Reconstructive Surgery, Chang Gung Memorial Hospital, Chang Gung University, College of Medicine , Kwei-San, Taoyuan 333, Taiwan Republic of China.
authors.forename: Chih-Hao
authors.initials: CH
authors.lastname: Chen
authors.affiliation: Department of Chemical and Materials Engineering, Chang Gung University , Kwei-San, Taoyuan 333, Taiwan Republic of China.
authors.forename: Jyh-Ping
authors.initials: JP
authors.lastname: Chen
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology*; D001861:Bone Regeneration / Q000187:drug effects*; D002113:Calcification, Physiologic / Q000187:drug effects; D002118:Calcium / Q000032:analysis; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D048271:Chitosan / Q000494:pharmacology*; D024042:Collagen Type I / Q000378:metabolism; D017886:Durapatite / Q000737:chemistry*; D005260:Female; D005349:Fibroins / Q000494:pharmacology*; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D008567:Membranes, Artificial; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D008819:Mice, Nude; D057139:Nanofibers / Q000737:chemistry* / Q000648:ultrastructure; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009928:Organ Specificity; D015675:Osteocalcin / Q000378:metabolism; D013052:Spectrometry, X-Ray Emission; D013194:Staining and Labeling
chemical_list: D055396:Bone Morphogenetic Protein 2; D024042:Collagen Type I; D008567:Membranes, Artificial; D015675:Osteocalcin; D005349:Fibroins; D048271:Chitosan; D017886:Durapatite; D000469:Alkaline Phosphatase; D002118:Calcium
keywords: bone morphogenetic protein; bone tissue engineering; chitosan; coaxial electrospinning; core−shell nanofibers; silk fibroin