Arpornmaeklong P, Suwatwirote N, Pripatnanont P, Oungbho K
Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Prince of Songkla University, Hat Yai, 90112 Songkhla, Thailand.
Int J Oral Maxillofac Surg. 2007 Apr;36(4):328-37. doi: 10.1016/j.ijom.2006.09.023. Epub 2007 Jan 12.
The aim of this study was to investigate the effects of collagen on the microstructure and biocompatibility of chitosan-collagen composite sponges fabricated by a freezing and drying technique. The study was categorized into four groups: Group I: collagen; Group II: chitosan; Group III: 1:1 (by wt) chitosan-collagen and Group IV: 1:2 (by wt) chitosan-collagen sponges. A mouse osteoblast cell line, MC3T3-E1, was cultivated on the sponges in a mineralized culture medium for 21 days. Microstructure of scaffolds and growth of cells on the sponges were examined using scanning electron and confocal laser scanning electron microscopes. Pore size was analysed from scanning electron microscope images using Image-Pro Plus image analysis software. Cell viability (MTT assay), alkaline phosphatase activity and levels of osteocalcin and calcium were monitored every 3 days and on days 15 and 21, respectively. It was found that the sponges were porous with average pore sizes of 80-100 microm. A combination of chitosan and collagen matrixes created a well defined porous microstructure and biocompatible scaffolds. Chitosan-collagen composite sponges promoted growth and differentiation of osteoblasts into the mature stage. To optimize application of the composite sponges in bone regeneration, the fabrication process must be improved to increase the pore size of the scaffolds.
本研究的目的是调查胶原蛋白对采用冷冻干燥技术制备的壳聚糖 - 胶原蛋白复合海绵微观结构和生物相容性的影响。该研究分为四组：第一组：胶原蛋白；第二组：壳聚糖；第三组：1:1（重量比）壳聚糖 - 胶原蛋白；第四组：1:2（重量比）壳聚糖 - 胶原蛋白海绵。将小鼠成骨细胞系MC3T3 - E1在矿化培养基中的海绵上培养21天。使用扫描电子显微镜和共聚焦激光扫描电子显微镜检查海绵支架的微观结构和细胞生长情况。使用Image - Pro Plus图像分析软件从扫描电子显微镜图像分析孔径。每3天以及分别在第15天和第21天监测细胞活力（MTT法）、碱性磷酸酶活性以及骨钙素和钙的水平。结果发现，海绵具有多孔结构，平均孔径为80 - 100微米。壳聚糖和胶原蛋白基质的组合形成了结构明确的多孔微观结构和生物相容性支架。壳聚糖 - 胶原蛋白复合海绵促进成骨细胞生长并分化至成熟阶段。为优化复合海绵在骨再生中的应用，必须改进制备工艺以增加支架的孔径。
