Liu Yan, Fu Yu, Liu Shuai, Zhou Yan-heng
Department of Orthodontics, Peking University School and Hospital of Stomatology, Beijing 100081, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2014 Feb 18;46(1):19-24.
To evaluate the effects of the microstructure of mineralized collagen scaffolds on cell morphology of MG 63.
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).
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.
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.
评估矿化胶原支架的微观结构对MG 63细胞形态的影响。
分别采用传统矿化方法和仿生自下而上矿化方法制备纤维外矿化胶原（EMC）支架和纤维内矿化胶原（IMC）支架。采用扫描电子显微镜（SEM）检查矿化胶原支架的微观结构及细胞与支架的相互作用。通过激光扫描显微镜（LSM）分析矿化方法对细胞黏附于胶原支架表面的影响。
通过SEM成像，两种矿化胶原支架呈现出不同的微观结构，包括胶原纳米纤维中磷灰石的大小、形态和位置。在EMC支架中，花状聚集体随机沉积在胶原纳米纤维周围，而纳米纤维表面未观察到磷灰石。使用与SEM联用的能量色散X射线光谱（EDS）证实了IMC支架中存在纤维内磷灰石矿相，钙磷比约为1.48。这种化学成分与天然骨组织相似。LSM结果表明，与EMC支架相比，IMC支架可促进细胞铺展。此外，在IMC支架上培养的细胞（18.54±2.71）与在EMC支架上培养的细胞（14.29±1.32）相比，纽蛋白染色密度更高。从SEM检查来看，两种矿化胶原支架均表现出良好的生物相容性。然而，细胞在不同支架上呈现出不同的形态。
矿化胶原支架的微观结构可影响细胞的初始黏附和形态。此外，IMC支架可促进细胞黏附和铺展。本研究将有助于我们制备用于牙槽骨再生的新型仿生材料。
