两种矿化胶原的显微结构对成骨样细胞MG 63形貌的影响

摘要/Abstract

摘要： 目的：研究两种矿化胶原的显微结构对人成骨样细胞MG 63的黏附力和形貌的影响。方法：采用传统矿化法和生物仿生矿化法分别制备纤维外矿化胶原（extrafibrillarly-mineralized collagen, EMC）与纤维内矿化胶原（intrafibrillarly-mineralized collagen, IMC）支架材料，人成骨样细胞MG 63作为实验用细胞，用扫描电子显微镜（scanning electron microscopy，SEM）检测两种支架材料的显微结构及细胞与材料间相互作用。采用激光共聚焦显微镜（laser scanning microscope，LSM）检测接种在材料上细胞的黏附力与形貌。结果：不同矿化方式影响胶原的显微结构。EMC组可见花样团簇状的晶体无规则沉积在纳米纤维束表面，而IMC组纤维束表面光滑，无晶体沉积。能谱分析结果显示其内部含有晶体，为含碳的钙缺乏型羟基磷灰石，这与天然骨矿化胶原成分类似。LSM结果显示，IMC比EMC支架材料更能促进MG 63细胞伸展。荧光定量分析结果表明MG 63细胞对IMC支架材料的黏附力（18.54 ± 2.71）明显高于EMC支架材料（14.29 ± 1.32）。SEM结果显示两种矿化胶原均具有良好的生物相容性，细胞在不同支架上呈现不同的形貌。结论：矿化胶原的显微结构影响细胞的形貌，IMC支架材料更能促进MG 63细胞的黏附与伸展，这将有助于研制新的牙槽骨再生的生物仿生材料。

关键词: 矿物质, 胶原, 组织支架, 细胞黏附, 骨再生

Abstract: Objective：To evaluate the effects of the microstructure of mineralized collagen scaffolds on cell morphology of MG 63. Methods:The extrafibrillarly mineralized collagen (EMC) and intrafibrillarly-mineralized collagen (IMC) scaffolds were fabricated separately by a conventional mineralization approach and a biomimetic, bottom-up mineralization approach. Scanning electron microscopy (SEM) was employed to examine the microstructure of the mineralized collagen scaffolds and cell-scaffold interactions. The effects of the mineralization methods on cell adhesion to the surface of the collagen scaffolds were analyzed by laser scanning microscope (LSM). Results:The two mineralized collagen scaffolds exhibited different microstructures, including the size, morphology and location of the apatites in collagen nanofibers by SEM imaging. In the EMC scaffold, flower-like aggregates randomly deposited around the collagen nanofibers, while no apatite was observed on the surface of the nanofibers. The presence of an intrafibrillar apatite mineral phase in the IMC scaffold was confirmed using energy dispersive X-ray spectroscopy (EDS) coupled to SEM, with the Ca ∶P ratio of approximately 1.48.This chemical composition was similar to natural bone tissue. The LSM results showed that the IMC scaffold could promote cell spreading compared with the EMC scaffold. Furthermore, the cells cultured on the IMC scaffold (18.54±2.71) showed higher density of vinculin staining than those on the EMC scaffold (14.29±1.32). From the SEM examination, both mineralized collagen scaffolds showed good biocompatibility. However, the cells exhibited different morphology on different scaffolds. Conclusion:The microstructure of the mineralized collagen scaffolds can affect the initial cell adhesion and morphology. Furthermore, the IMC scaffold can promote cell adhesion and spreading. The present study will help us to fabricate novel biomimetic materials for alveolar bone regeneration.

Key words: Mineralized collagen, Scaffold, Cell adhesion, Cell morphology, Alveolar bone regeneration

刘燕，付玉，刘帅，周彦恒. 两种矿化胶原的显微结构对成骨样细胞MG 63形貌的影响[J]. 北京大学学报（医学版）, 2014, 46(1): 19-24.

LIU Yan, FU Yu, LIU Shuai, ZHOU Yan-heng. Effects of microstructure of mineralized collagen scaffolds on cell morphology of MG 63[J]. Journal of Peking University(Health Sciences), 2014, 46(1): 19-24.

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