Key Laboratory for Oral Biomedical Engineering of Ministry of Education, School and Hospital of Stomatology, Wuhan University, Wuhan, 430079, People's Republic of China.
J Mater Sci Mater Med. 2010 Feb;21(2):489-96. doi: 10.1007/s10856-009-3931-x. Epub 2009 Nov 12.
The development of suitable bioactive three-dimensional scaffold for the promotion of cellular proliferation and differentiation is critical in periodontal tissue engineering. In this study,porous beta-tricalcium phosphate/chitosan composite scaffolds were prepared through a freeze-drying method. These scaffolds were evaluated by analysis of microscopic structure, porosity, and cytocompatibility. The gene expression of bone sialoprotein (BSP) and cementum attachment protein (CAP) was detected with RT-PCR after human periodontal ligament cells (HPLCs) were seeded in these scaffolds. Then cell-scaffold complexes were implanted subcutaneously into athymic mice. The protein expression of alkaline phosphatase (ALP) and osteopontin (OPN) was detected in vivo. Results indicated that composite scaffolds displayed a homogeneous three-dimensional microstructure; suitable pore size (120 microm) and high porosity (91.07%). The composite scaffold showed higher proliferation rate than the pure chitosan scaffold, and up-regulated the gene expression of BSP and CAP. In vivo, HPLCs in the composite scaffold not only proliferated but also recruited vascular tissue ingrowth. The protein expression of ALP and OPN was up-regulated in the composite scaffold. Therefore, it was suggested that the composite scaffold could promote the differentiation of HPLCs towards osteoblast and cementoblast phenotypes.
在牙周组织工程中,开发合适的具有生物活性的三维支架以促进细胞增殖和分化是至关重要的。本研究通过冷冻干燥法制备了多孔β-磷酸三钙/壳聚糖复合支架。通过分析微观结构、孔隙率和细胞相容性对这些支架进行了评估。将人牙周膜细胞(HPLCs)接种到这些支架上后,通过 RT-PCR 检测骨涎蛋白(BSP)和牙骨质附着蛋白(CAP)的基因表达。然后将细胞-支架复合物皮下植入无胸腺小鼠体内。体内检测碱性磷酸酶(ALP)和骨桥蛋白(OPN)的蛋白表达。结果表明,复合支架呈现均匀的三维微观结构;合适的孔径(120 微米)和高孔隙率(91.07%)。与纯壳聚糖支架相比,复合支架显示出更高的增殖率,并上调了 BSP 和 CAP 的基因表达。在体内,复合支架中的 HPLCs 不仅增殖,而且还招募了血管组织的生长。复合支架中 ALP 和 OPN 的蛋白表达上调。因此,该复合支架可促进 HPLCs 向成骨细胞和成牙骨质细胞表型分化。