Shin Yong Cheol, Lee Jong Ho, Jin Linhua, Kim Min Jeong, Kim Chuntae, Hong Suck Won, Oh Jin Woo, Han Dong-Wook
J Nanosci Nanotechnol. 2015 Oct;15(10):7907-12. doi: 10.1166/jnn.2015.11214.
Recently, there has been considerable effort to develop suitable scaffolds for tissue engineering applications. Cell adhesion is a prerequisite for cells to survive. In nature, the extracellular matrix (ECM) plays this role. Therefore, an ideal scaffold should be structurally similar to the natural ECM and have biocompatibility and biodegradability. In addition, the scaffold should have biofunctionality, which provides the potent ability to enhance the cellular behaviors, such as adhesion, proliferation and differentiation. This study concentrates on fabricating cell-adhesive matrices composed of RGD peptide-displaying M13 bacteriophage (RGD-M13 phage) and poly(lactic-co-glycolic acid, PLGA) nanofibers. Long rod-shaped M13 bacteriophages are non-toxic and can express many desired proteins on their surface. A genetically engineered M13 phage was constructed to display RGD peptides on its surface. PLGA is a biodegradable polymer with excellent biocompatibility and suitable physicochemical property for adhesive matrices. In this study, RGD-M13 phage/PLGA hybrid nanofiber matrices were fabricated by electrospinning. The physicochemical properties of these matrices were characterized by scanning electron microscopy, atomic force microscopy, Raman spectroscopy, and contact angle measurement. In addition, the cellular behaviors, such as the initial attachment, proliferation and differentiation, were analyzed by a CCK-8 assay and immunofluorescence staining to evaluate the potential application of these matrices to tissue engineering scaffolds. The RGD-M13 phage/PLGA nanofiber matrices could enhance the cellular behaviors and promote the differentiation of C2C12 myoblasts. These results suggest that the RGD-M13 phage/PLGA nanofiber matrices are beneficial to myoblast differentiation and can serve as effective tissue engineering scaffolds.
近年来,人们付出了巨大努力来开发适用于组织工程应用的支架。细胞黏附是细胞存活的先决条件。在自然界中,细胞外基质(ECM)发挥着这一作用。因此,理想的支架在结构上应与天然ECM相似,并具有生物相容性和生物可降解性。此外,支架应具有生物功能性,即具备增强细胞行为(如黏附、增殖和分化)的强大能力。本研究专注于制备由展示RGD肽的M13噬菌体(RGD-M13噬菌体)和聚乳酸-乙醇酸共聚物(PLGA)纳米纤维组成的细胞黏附基质。长杆状的M13噬菌体无毒,且能在其表面表达多种所需蛋白质。构建了一种基因工程M13噬菌体,使其在表面展示RGD肽。PLGA是一种生物可降解聚合物,具有优异的生物相容性和适合黏附基质的物理化学性质。在本研究中,通过静电纺丝制备了RGD-M13噬菌体/PLGA混合纳米纤维基质。通过扫描电子显微镜、原子力显微镜、拉曼光谱和接触角测量对这些基质的物理化学性质进行了表征。此外,通过CCK-8测定法和免疫荧光染色分析了细胞行为,如初始附着、增殖和分化,以评估这些基质在组织工程支架中的潜在应用。RGD-M13噬菌体/PLGA纳米纤维基质可增强细胞行为并促进C2C12成肌细胞的分化。这些结果表明,RGD-M13噬菌体/PLGA纳米纤维基质有利于成肌细胞分化,可作为有效的组织工程支架。