胰岛素样生长因子1、血小板源性生长因子、转化生长因子β1对人退变纤维环细胞生物学活性的影响  

Effects of insulin-like growth factor-1, platelet-derived growth factor and transforming growth factor-beta 1 on biologic activity of human degenerative annulus fibrosus cells

作　　者：林旺[1] 王万明[1] 庄颜峰[1] 魏梅洋[1] 李杰[1] 林增平[1] 

机构地区：[1]解放军南京军区福州总医院骨二科,福建省福州市350025

出　　处：《中国组织工程研究与临床康复》2009年第37期7309-7313,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：南京军区医药卫生科研资助项目~~

摘　　要：背景:生长因子可调整椎间盘细胞的增殖、分化及基质代谢,用于椎间盘退变的生物学治疗,既往的研究偏重于成骨性生长因子对髓核细胞的作用,对纤维环细胞的研究相对较少。目的:观察体外培养条件下胰岛素样生长因子1、血小板源性生长因子、转化生长因子β1对人退变纤维环细胞生物学活性的影响。设计、时间及地点:对照观察实验,于2007-11/2008-12在解放军南京军区福州总医院完成。材料:腰椎间盘手术患者的纤维环组织。方法:结合酶消化法和组织块培养法,体外单层培养人退变纤维环原代细胞。传代后通过免疫组织化学染色鉴定细胞。对传3代细胞分别采用不同生长因子干预,分为100μg/胰岛素样生长因子1组、10μg/L血小板源性生长因子组、10μg/转化生长因子β1组、100μg/L胰岛素样生长因子1+10μg/L血小板源性生长因子组、100μg/L胰岛素样生长因子1+10μg/L转化生长因子β1组、10μg/L血小板源性生长因子+10μg/L转化生长因子β1组、100μg/L胰岛素样生长因子1+10μg/L血小板源性生长因子+10μg/L转化生长因子β1组,以不加生长因子为对照组。主要观察指标:免疫组织化学染色观察传3代细胞。干预3,6d后,采用MTT法测定传3代细胞增殖,ELISA法测定细胞培养上清液中Ⅰ、Ⅰ型胶原和聚集蛋白聚糖质量浓度。结果:传3代细胞Ⅰ、Ⅰ型胶原免疫组织化学染色阳性。胰岛素样生长因子1促进人退变纤维环细胞增殖,轻度抑制细胞合成Ⅰ型胶原,促进合成Ⅰ型胶原和聚集蛋白聚糖,促Ⅰ型胶原合成作用轻度强于转化生长因子β1。血小板源性生长因子促进细胞增殖,作用强于胰岛素样生长因子1,其抑制细胞合成Ⅰ型胶原,轻度促进合成Ⅰ型胶原,无促合成聚集蛋白聚糖作用。转化生长因子β1抑制细胞增殖,促进合成Ⅰ、Ⅰ型胶原及聚集蛋白聚糖,促聚集蛋白聚糖合成作用强于胰岛素样�BACKGROUND： Growth factors are usually used for the biological treatment of degenerative intervertebral disc （IVD）, for they can contribute to the regulation of the proliferation, differentiation and matrix metabolism of IVD cells. However, previous studies focused more on effects of osteogenic growth factors on nucleus pulposus cells, instead of annulus fibrosus （AF） cells. OBJECTIVE： To study the in vitro effects of insulin-like growth factor-1（IGF-1）, platelet-derived growth factor （PDGF） and transforming growth factor-β1（TGF-β1） on the biologic activity of human degenerative AF cells. DESIGN, TIME AND SETTING： A controlled observational experiment was carried out in Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA between November 2007 and December 2008. MATERIALS： AF tissues were obtained from patients with lumbar disc surgeries. METHODS： Primary cells of human degenerative AF were obtained through in vitro monolayer culture combined with enzyme digestion method and tissue pieces culture method. After passage, cells were identified through immunohistochemical staining. Cells of passage 3 were treated separately with different growth factors .They were divided into many groups as following： 100 μg/L IGF-1 group, 10 μg/L PDGF group, 10 μg/L TGF-β1 group, 100 μg/L IGF-1＋10 μg/L PDGF group, 100 μg/L IGF-1＋ 10 μg/L TGF-β1 group, 10 μg/L PDGF＋10 μg/L TGF-β1 group, 100 μg/L IGF-1＋10 μg/L PDGF＋10 μg/L TGF-β1 group and the control group without any growth factor. MAIN OUTCOME MEASURES： Cells of passage 3 were identified by immunohistochemical staining. At day 3 and day 6 post the treatment with growth factors ,the cell proliferation was detected by MTT assay; the concentration of collagen typeⅠ, typeⅡand aggrecan in the cell culture supernatant were detected by enzyme-linked immunosorbent assay （ELISA）. RESULTS： Collagens typeⅠand typeⅡ immunohistochemical staining of passage 3 cells were positive. IGF-1 had the effects

关 键 词：胰岛素样生长因子1 血小板源性生长因子 转化生长因子Β1 纤维环细胞 

分 类 号：R392.114[医药卫生—免疫学]