壳聚糖支架复合重组人成骨蛋白1对体外培养兔髓核和纤维环细胞蛋白聚糖合成的影响  

Chitosan scaffold combined with recombinant human osteogenic protein-1 promotes proteoglycan synthesis of rabbit anulus fibrosus and nucleus pulposus cells

作　　者：田海泉 李放[2] 马舟涌[2] 陈晓斌[2] 王晓伟 

机构地区：[1]西医科大学第二临床医院骨科,山西省太原市030001 [2]解放军北京军区总医院全军骨科研究所,北京市100700

出　　处：《中国组织工程研究与临床康复》2008年第27期5209-5213,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:成骨蛋白1有效刺激软骨细胞蛋白聚糖和胶原蛋白合成已被证实,椎间盘内注射成骨蛋白1可增加椎间盘高度也在动物模型中得到证实。目的:体外评价重组人成骨蛋白1对培养于壳聚糖凝胶中的兔椎间盘细胞增殖和蛋白聚糖合成的作用。设计、时间及地点:成组设计,实验于2007-08/2008-02在北京军区总医院骨科实验室完成。材料:清洁级4月龄日本大耳兔5只。重组人成骨蛋白1为Sigma分装。医用级壳聚糖,脱乙酰度95.11%,黏度155mPa.s,Mr870000,由北京纵横洋洲医药生物科技有限公司提供。方法:无菌条件下取日本大耳兔椎间盘10个,分离其髓核和纤维环细胞并接种于自制壳聚糖凝胶支架,在培养液中加入重组人成骨蛋白1,并分为3组:对照组(无重组人成骨蛋白1)、100μg/L重组人成骨蛋白1组、200μg/L重组人成骨蛋白1组。主要观察指标:培养7,14,21d时对培养物DNA含量、蛋白聚糖合成量(35SO42-摄入量、硫酸盐黏多糖含量)进行检测。结果:100,200μg/L重组人成骨蛋白1组的DNA含量没有减少,对照组DNA含量逐渐下降。100,200μg/L重组人成骨蛋白1组蛋白聚糖合成量明显高于对照组,200μg/L重组人成骨蛋白1组升高更为显著。100,200μg/L重组人成骨蛋白1组在3个时间点均呈现上升趋势,培养14d和21d的差异有显著性意义(P<0.001)。结论:重组人成骨蛋白1可有效促进体外培养的兔髓核和纤维环细胞增殖及蛋白聚糖合成。BACKGROUND： Osteogenic protein-1 （OP-1） can effectively stimulate the proteoglycan and collagen synthesis by articular chondrocytes, and a single injection of OP-1 into intervertebral disc has been proved to raise the disc height in animal models. OBJECTIVE： To in vitro determine the effect of recombinant human OP-1 （rhOP-1） on stimulating the cell proliferation and proteoglycan metabolism of rabbit intervertebral disc cells cultured in chitosan-gel. DESIGN, TIME AND SETTING： The experiments at grouping design were carried out at the Orthopaedic Laboratory in General Hospital of Beijing Military Area Command of Chinese PLA （Beijing, China） from August 2007 to February 2008. MATERIALS： Five Japanese rabbits of clean grade and four months old, were adopted in this study. The rhOP-1 was a product of Sigma Company （USA） in divided package. Medical chitosan, deacetylation degree 95.11%, viscosity mPa.s, M~=870 000, was offered by Beijing Transglobal Biomedical Technology Co,. Ltd （China）. METHODS： Ten rabbit intervertebral discs were harvested, then nucleus pulposus and anulus fibrosus cells were isolated from the discs and inoculated on the self-made chitosan-gel scaffold. Culture solution was divided into three groups： control group without rhOP-1,100 μ g/L rhOP-1 group and 200 μg/L rhOP-1 group. MAIN OUTCOME MEASURES： The synthesis and accumulation of proteoglycans （35SO4^2- intake and sulfate glycosaminoglycan content） and the DNA content were biochemically assessed at days 7, 14 and 21 of the culture. RESULTS： The DNA content was unchanged in two rhOP-1 groups, while that in the control group was reduced. Treatment with rhOP-1, compared with the control condition, significantly upregulated proteoglycan synthesis and accumulation in all cases tested. A longer exposure over 14 days to rhOP-1 resulted in a pronounced response and there were significant differences between day 14 and day 21 （P 〈 0.001）. CONCLUSION： The rhOP-1 is effective to promote the cell prolifera

关 键 词：髓核细胞 纤维环细胞 成骨蛋白1 壳聚糖 蛋白聚糖 生物材料 

分 类 号：R318[医药卫生—生物医学工程]