Zhang Yu-Feng, Cheng Xiang-Rong, Chen Yun, Shi Bin, Chen Xiao-Hui, Xu Dong-Xuan, Ke Jin
Ministry Education Key Laboratory for Oral Biomedical Engineering School of Stomatology, Wuhan University, Wuhan 430079, PR China.
J Biomater Appl. 2007 Apr;21(4):333-49. doi: 10.1177/0885328206063853. Epub 2006 Mar 16.
The development of suitable three-dimensional scaffold for the maintenance of cellular viability and differentiation is critical for applications in periodontal tissue engineering. In this work, different ratios of porous nanohydroxyapatite/chitosan (HA/chitosan) scaffolds are prepared through a freeze-drying process. These scaffolds are evaluated in vitro by the analysis of microscopic structure, porosity, and cytocompatibility. The expression of type I collagen and alkaline phosphatase (ALP) activity are detected with real-time polymerase chain reaction (RT-PCR). Human periodontal ligament cells (HPLCs) transfected with enhanced green fluorescence protein (EGFP) are seeded onto the scaffolds, and then these scaffolds are implanted subcutaneously into athymic mice. The results indicated that the porosity and pore diameter of the HA/chitosan scaffolds are lower than those of pure chitosan scaffold. The HA/chitosan scaffold containing 1% HA exhibited better cytocompatibility than the pure chitosan scaffold. The expression of type I collagen and ALP are up-regulated in 1% HA/chitosan scaffold. After implanted in vivo, EGFP-transfected HPLCs not only proliferate but also recruit surrounding tissue to grow in the scaffold. The degradation of the scaffold significantly decreased in the presence of HA. This study demonstrated the potential of HA/ chitosan scaffold as a good substrate candidate in periodontal tissue engineering.
开发适用于维持细胞活力和分化的三维支架对于牙周组织工程应用至关重要。在这项工作中，通过冷冻干燥工艺制备了不同比例的多孔纳米羟基磷灰石/壳聚糖（HA/壳聚糖）支架。通过对微观结构、孔隙率和细胞相容性的分析对这些支架进行体外评估。用实时聚合酶链反应（RT-PCR）检测I型胶原蛋白的表达和碱性磷酸酶（ALP）活性。将转染了增强型绿色荧光蛋白（EGFP）的人牙周膜细胞（HPLCs）接种到支架上，然后将这些支架皮下植入无胸腺小鼠体内。结果表明，HA/壳聚糖支架的孔隙率和孔径低于纯壳聚糖支架。含1%HA的HA/壳聚糖支架比纯壳聚糖支架表现出更好的细胞相容性。I型胶原蛋白和ALP的表达在1%HA/壳聚糖支架中上调。体内植入后，转染EGFP的HPLCs不仅增殖，还能募集周围组织在支架中生长。在HA存在下，支架的降解显著降低。本研究证明了HA/壳聚糖支架作为牙周组织工程中良好候选基质的潜力。
