中国组织工程研究

中国组织工程研究

• 骨髓干细胞 • 上一篇    下一篇

离心力对兔骨髓基质细胞成骨分化的影响

徐南伟，张 煜，周 栋，孙荣彬   

  1. 南京医科大学附属常州市第二人民医院骨科，江苏省常州市 213003
  • 出版日期:2010-01-04 发布日期:2010-01-04
  • 作者简介:徐南伟，男，1963年生，江苏省常州市人，汉族，1985年苏州医学院毕业，主任医师，副教授，主要从事脊柱外科、计算机辅助骨科及骨组织工程方面的研究。 xunanwei1963@yahoo.com.cn

Effect of centrifugal force on osteoblastic differentiation of bone marrow stroma cells in vitro

Xu Nan-wei, Zhang Yu, Zhou Dong, Sun Rong-bin   

  1. Department of Orthopedics, Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou   213003, Jiangsu Province, China
  • Online:2010-01-04 Published:2010-01-04
  • About author:Xu Nan-wei, Chief physician, Associate professor, Department of Orthopedics, Changzhou Second People’s Hospital of Nanjing Medical University, Changzhou 213003, Jiangsu Province, China xunanwei1963@yahoo.com.cn

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416

被引次数

3

摘要：

背景：在骨髓基质细胞向成骨细胞分化过程中,离心力是一种促进因素。

目的：探讨离心力是否能促进复合于聚羟基乙酸共聚物材料上的兔骨髓基质细胞向成骨方向分化。

方法：全骨髓法分离培养兔骨髓基质细胞，贴壁法纯化，至细胞达80%融合时胰酶-EDTA消化传代，调整细胞浓度为1×    109 L-1。将聚羟基乙酸共聚物切割成5 mm×5 mm大小，预先浸泡入含血清培养基内24 h，然后将第3代骨髓基质细胞悬液按300 μL/块密度接种于支架材料表面，将复合物置入离心管底，细胞滴加面朝上，加入含抗坏血酸、β-甘油磷酸钠、地塞米松的成骨诱导培养基。设立2组：离心力刺激组每间隔12 h将离心培养管置于离心机上，按1 000 r/min离心30 min，相对离心力132 g；对照组培养管中细胞不作离心静态培养。通过光镜观察及检测碱性磷酸酶活性、骨钙素含量、组织钙含量，评估成骨细胞的分化水平。

结果与结论：体外培养第16天，离心力刺激组支架表面被多层细胞和矿化基质所覆盖，而对照组仅支架表层见一薄层细胞。与对照组比较，离心力刺激组培养后第2天碱性磷酸酶活性明显降低(P < 0.05)，第4天碱性磷酸酶活性明显升高(P < 0.05)。在整个培养期间对照组骨钙素质量浓度一直维持低水平，离心力刺激组第12，16天骨钙素质量浓度明显高于对照组(P < 0.05)。培养16 d后离心力刺激组的钙质量浓度明显高于对照组(P < 0.05)。提示离心力能促进接种于聚羟基乙酸共聚物的兔骨髓基质细胞成骨分化，并形成矿化产物。

关键词: 离心力, 骨髓基质细胞, 成骨分化, 组织工程, 兔

Abstract:

BACKGROUND: Centrifugal force is a contributing factor inducing osteoblastic differentiation from bone marrow stroma cells.

OBJECTIVE: To explore whether centrifugal force promote osteoblastic differentiation from rabbit marrow stroma cell seeded on polylactic-co-glycolic acid (PLGA) scaffolds.

METHODS: Rabbit bone marrow stroma cells were isolated and cultured by whole bone marrow method, purified by attachment method, and digested by trypsin-EDTA at 80% confluency. The cell concentration was adjusted to 1×109/L. PLGA was cut into pieces, 5 mm×5 mm, soaked in serum-conditioned culture medium for 24 hours. The third passage of bone stroma cell suspension at a density of 300 μL was respectively seeded into the PLGA material. The scaffold/cell compound was placed in centrifuge tube, with cell at the upper layer and cultured in osteoblastic induced medium containing antiscorbic acid, β-sodium glycerophosphate, and dexamethasone respectively under centrifugal force every 12 hours (1 000 r/min for 30 minutes, relative centrifugal force 132 g) and static condition. Alkaline phosphatase activity, osteocalcin content and calcium content as well as observation by light microscopy were used to evaluate osteoblastic differentiation.

RESULTS AND CONCLUSION: After 16 days of in vitro culture, the scaffolds of centrifugal force group were coated by multiplayer cells and mineralized matrix, but only a thin layer of cells were observed on the scaffold of control group. The centrifugal force system resulted in a significant decrease in alkaline phosphatase activity at day 2 (P < 0.05) but significant increase at day 4 compared with the static culture condition (P < 0.05). During the whole culture time, osteocalcin secretion remained low in control group. At days 12 and 16, a significant enhancement in osteocalcin secretion was observed for centrifugal force culture compared with static culture conditions (P < 0.05). Moreover, after 16 days of culture a significant increase in calcium deposition was observed in the scaffolds subjected to centrifugal force compared with static culture condition (P < 0.05). Centrifugal force can enhance osteoblastic differentiation and mineralized matrix production of bone marrow stroma cell seeded in PLGA.

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