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尼古丁对兔骨髓基质干细胞增殖及向软骨细胞方向分化的影响
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作者Author单位UnitE-Mail
应晓洲 YING Xiao-zhou 温州医学院附属第二医院创伤骨科,浙江 温州 325000 Department of Orthopaedics Surgery, the Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang, China  
彭磊 PENG Lei 温州医学院附属第二医院创伤骨科,浙江 温州 325000
海南医学院附属医院创伤急救中心
Department of Orthopaedics Surgery, the Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang, China xiaobo197518@163.com 
程少文 CHENG Shao-wen 温州医学院附属第二医院创伤骨科,浙江 温州 325000 Department of Orthopaedics Surgery, the Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang, China  
陈庆玉 CHEN Qing-yu 温州医学院附属第二医院创伤骨科,浙江 温州 325000 Department of Orthopaedics Surgery, the Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang, China  
张伟 ZHANG Wei 温州医学院附属第二医院创伤骨科,浙江 温州 325000 Department of Orthopaedics Surgery, the Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang, China  
寇冬泉 KOU Dong-quan 温州医学院附属第二医院创伤骨科,浙江 温州 325000 Department of Orthopaedics Surgery, the Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang, China  
沈跃 SHEN Yue 温州医学院附属第二医院创伤骨科,浙江 温州 325000 Department of Orthopaedics Surgery, the Second Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, Zhejiang, China  
期刊信息:《中国骨伤》2011年24卷,第11期,第935-938页
DOI:10.3969/j.issn.1003-0034.2011.11.015


目的:探讨不同浓度尼古丁对单层培养的骨髓基质干细胞及向软骨方向分化的影响。

方法:获取兔骨髓基质干细胞,倒置显微镜下观察细胞形态。取第3代细胞,加入不同浓度(0、1×10-7、1×10-6、1×10-5 M)的尼古丁,分别于1、4、7、14 d用MTT法检测其对骨髓基质干细胞增殖的影响。取第3代细胞进行诱导分化,用逆转录聚合酶链式反应(RT- PCR )法检测Ⅱ型胶原和蛋白聚糖的表达。

结果:镜下见细胞形态由圆形向梭形转化;1×10-7、 1×10-6 M尼古丁浓度能够促进骨髓基质干细胞的增殖,1×10-5 M尼古丁则会抑制骨髓基质干细胞的增殖活性。1×10-7、 1×10-6 M(特别是1×10-6 M)的尼古丁能够增加诱导后Ⅱ型胶原的表达,并随时间延长而增加。而1×10-7 M尼古丁在第7天时能促进诱导后蛋白聚糖的表达,1×10-5 M尼古丁则能抑制Ⅱ型胶原及蛋白聚糖mRNA 表达。

结论:低浓度的尼古丁能够促进骨髓基质干细胞的增殖及向软骨细胞方向分化,高浓度的尼古丁则对其有抑制作用,由此推测在软骨组织工程中,局部适时适量的应用尼古丁是一种能够促进骨髓基质干细胞向软骨细胞方向分化的有前景的治疗方法。
[关键词]:干细胞,骨髓  尼古丁  胶原Ⅱ型  蛋白聚糖类  软骨
 
Effects of nicotine on bone marrow stromal cells proliferation and differentiation of chondrocyte in vitro
Abstract:

Objective: To examine the effects of various concentration of nicotine on bone marrow stromal cells (BMSCs) proliferation and differentiation of cartilaginous in vitro.

Methods: BMSCs was obtained from femoral bone and tibia of New-Zealand albino rabbit. The cells of the 3rd generation were used in study. Different concentration of nicotine(0, 1×10-7, 1×10-6, 1×10-5 M) were added into BMSCs. BMSCs proliferation was analyzed by MTT assay at the 1, 4, 7, 14 days. The expression of collagen typeⅡand aggrecan as the marker genes of cartilaginous differentiation from BMSCs were detected by reverse transcriptase-polymerase chain reaction(RT-PCR).

Results: Microscope showed that BMSCs transformed from round to fusiform shape. The concentration of nicotine in 1×10-7, 1×10-6 M had a significant positive effect on cell proliferation and the expression of typeⅡcollagen in a time-dependent manner when supplemented in commonly used induction media(P<0. 05). Concentrations of nicotine in 1×10-7 can promote the expression of aggrecan at the 7th day after induction, and in 1×10-5 M may inhibit the expression of typeⅡcollagen and aggrecan.

Conclusion: It was implied that local application of nicotine at an appropriate concentration may be a promising approach for enhancing cartilaginous differentiation capacity of BMSCs in cartilage tissue engineering.
KEYWORDS:Stem cells, myeloid  Nicotine  Collagen typeⅡ  Proteoglycans  Cartilage
 
引用本文,请按以下格式著录参考文献:
中文格式:应晓洲,彭磊,程少文,陈庆玉,张伟,寇冬泉,沈跃.尼古丁对兔骨髓基质干细胞增殖及向软骨细胞方向分化的影响[J].中国骨伤,2011,24(11):935~938
英文格式:YING Xiao-zhou,PENG Lei,CHENG Shao-wen,CHEN Qing-yu,ZHANG Wei,KOU Dong-quan,SHEN Yue.Effects of nicotine on bone marrow stromal cells proliferation and differentiation of chondrocyte in vitro[J].zhongguo gu shang / China J Orthop Trauma ,2011,24(11):935~938
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