外源性Ⅰ型胶原对人胚骨膜成骨细胞生物学特性的影响

The Influence of Type Ⅰ Collagen on the Cell Behavior o f Human Embryoni c Periosteous Osteoblasts

目的 研究 型胶原对人胚骨膜来源成骨细胞生物学特性的影响 ,为 型胶原在组织工程人工骨中的应用提供依据。方法 在不同浓度 型胶原涂层上培养成骨细胞 ,用细胞计数法研究成骨细胞粘附情况 ,3 H- Td R掺入实验观察成骨细胞增殖能力 ,通过检测成骨细胞胶原、骨钙素和碱性磷酸酶的合成情况 ,以不涂层 型胶原为对照组 ,比较骨膜来源成骨细胞成骨能力的改变。结果 型胶原对成骨细胞发挥以下作用 :1增加粘附细胞数量 ,且在 2 5 μg/ m l浓度达最大效应 ;2减弱成骨细胞增殖能力 ,且以 12 .5 μg/ ml以上浓度作用显著 ( P<0 .0 5 ) ;3轻度减弱成骨细胞 型胶原合成能力 ,以 2 5 μg/ ml以上浓度作用显著 ( P<0 .0 5 ) ;4增加骨钙素合成 ,以6 .2 5 μg/ m l以上浓度作用显著 ( P<0 .0 5 ) ,2 5 μg/ m l浓度达最大效应 ;5增加成骨细胞碱性磷酸酶活性 ,以 12 .5 μg/ml以上浓度作用显著 ( P<0 .0 5 )。结论 型胶原能促进成骨细胞的粘附与分化 ;支架材料上复合 型胶原涂层可增强成骨细胞的成骨能力 ; 型胶原最佳复合浓度为 2 5 μg/

Objective To study the influence of type Ⅰ coll agen(COL Ⅰ) on the cell behavior of human periosteous osteoblasts(OB) and the application of type Ⅰ collagen in constructing bioactive artifical bone.Methods OB were cultured on dishes coated with bovine type Ⅰ collage n in different final concentrations. The cell adhesion was examined by the meth o ds of cell count, the proliferation of OB was studied by 3H TdR, and the ost eoblastic ability was assessed by the synthesis of collagen, osteocalcin and alk aline phosphatase (ALP). Results OB cultured on type Ⅰ collage n layer had the following characteristicis: ① The amounts of adhesive cells we re maximal top in 25 μg/ml final concentration; ② The proliferation of OB was decreased above 12.5 μg/ml final concentration ( P <0.05); ③ The synthesis of type Ⅰ collagen was reduced slightly (above 25 μg/ml, P <0.05); ④ The secretion of osteocalcin was increased markedly (above 6.25 μg/ml, P <0.05, w hich reached maximally in 25μg/ml); ⑤ The ALP activity was also increased (ab ove 12.5μg/ml, P <0.05). Conclusion Type Ⅰ collagen promot es the expression of osteoblastic phenotype and cell adhesion. When the scaffold materials for bone tissue engineering are coated with type Ⅰ collagen, the ost eogenesis of OB is enhanced to accelerate the tranformation course from artifici al bone to biological bone, the best final concentration is 25μg/ml.