中国组织工程研究 ›› 2012, Vol. 16 ›› Issue (11): 1901-1905.doi: 10.3969/j.issn.1673-8225.2012.11.001

• 骨组织构建 bone tissue construction •    下一篇

下颌骨牵引成骨过程中基因干预对牵引区转化生长因子β1表达的影响?****

何小川,李绍兰,胡纯兵,刘  震,高志丹,尹  康,吴国平,郭  力   

  1. 泸州医学院附属医院整形外科,四川省泸州市  646000
  • 收稿日期:2011-07-30 修回日期:2011-10-11 出版日期:2012-03-11
  • 通讯作者: 吴国平,博士,主任医师,泸州医学院附属医院整形外科,四川省泸州市 646000 drgpwu@yahoo.com.cn
  • 作者简介:何小川,男,1964年生,四川省泸州市人,汉族,1989年华西医科大学毕业,副主任医师,主要从事整形外科基础与临床的研究。 lzmchxc@sina.com 并列第一作者:李绍兰★,女,1983年生,四川省泸州市人,汉族,2011年泸州医学院毕业,硕士,医师,主要从事整形外科基础与临床研究。 liyixing.good@163.com
  • 基金资助:

    国家自然科学基金资助项目(30600653);四川省应用基础研究计划项目(2006J13-128);四川省卫生厅科研课题(060044,100285)。

     

Effects of gene therapy on the expression of transforming growth factor beta 1 during mandible distraction osteogenesis   

He Xiao-chuan, Li Shao-lan, Hu Chun-bing, Liu Zhen, Gao Zhi-dan, Yin Kang, Wu Guo-ping, Guo Li   

  1. Department of Plastic Surgery, Affiliated Hospital of Luzhou Medical College, Luzhou  646000, Sichuan Province, China
  • Received:2011-07-30 Revised:2011-10-11 Online:2012-03-11
  • Contact: author: Wu Guo-ping, Doctor, Chief physician, Department of Plastic Surgery, Affiliated Hospital of Luzhou Medical College, Luzhou 646000, Sichuan Province, China drgpwu@yahoo.com.cn
  • About author:He Xiao-chuan, Associate chief physician, Department of Plastic Surgery, Affiliated Hospital of Luzhou Medical College, Luzhou 646000, Sichuan Province, China lzmchxc@sina.com Li Shao-lan★, Master, Physician, Department of Plastic Surgery, Affiliated Hospital of Luzhou Medical College, Luzhou 646000, Sichuan Province, China liyixing.good@163.com
  • Supported by:

    National Natural Science Foundation of China, No.30600653*; Projects of Basic Research Application in Sichuan Province, No.2006J13-128*; Research Projects of Sichuan Provincial Health Bureau, No.060044*, 100285*

摘要:

 

背景:局部基因治疗能促进牵引区新骨的生成,但关于基因治疗后对局部生长因子表达的影响目前尚不清楚。
目的:观察电穿孔介导的基因治疗对兔下颌骨牵引成骨过程中转化生长因子β1表达的影响。
方法:新西兰大白兔双侧下颌骨截骨后3 d开始下颌骨牵引,0.8 mm/d,连续牵引7 d后,随机分为5组,分别在牵引区注射2 μg(0.1 g/L)重组质粒pIRES-hVEGF165-hBMP2、pIRES-hBMP2、pIRES-hVEGF165、空质粒pIRES及相同剂量的生理盐水。之后施加电穿孔刺激。
结果与结论:免疫组织化学染色发现转化生长因子β1主要在细胞胞浆中表达,给药7 d时骨端骨细胞、编织骨痂骨细胞、骨痂表面成骨细胞呈转化生长因子β1染色阳性;14 d时新生成的编织骨痂骨细胞、骨痂表面成骨细胞、肉芽组织中的间质细胞、单核巨细胞、多核巨细胞转化生长因子β1染色阳性;28 d时转化生长因子β1阳性细胞明显减少。其中注射重组质粒pIRES-hVEGF165-hBMP2、pIRES-hBMP2、pIRES-hVEGF165后转化生长因子β1的表达明显多于注射空质粒pIRES及生理盐水(P < 0.05或P < 0.01)。说明基因治疗能促进转化生长因子β1的表达,促进牵引区细胞基质的形成和新骨生成。
 

关键词: 电穿孔, 基因疗法, 骨生成, 牵张, 转化生长因子&beta, 1

Abstract:

BACKGROUND: Although recent studies have shown that the local gene therapy can promote the formation of new bone in distraction gap, the effect of gene therapy on the expression of local growth factors is unclear.
OBJECTIVE: To investigate the effect of electroporation-mediated gene therapy on the expression of transforming growth factor-β1 (TGF-β1) during mandibular distraction osteogenesis in a rabbit model.
METHODS: Mandibular distraction of 0.8 mm/d was performed in New-Zealand rabbits at 3 days after bilateral mandibular osteotomy and lasted for 7 days. After the completion of distraction, the rabbits were randomly divided into five groups. 2 μg      (0.1 μg/μL) recombinant plasmid pIRES-hVEGF165-hBMP2, pIRES-hBMP2, pIRES-hVEGF165, pIRES and normal saline was injected into the distraction area respectively. After injection, every group employed electroporation.
RESULTS AND CONCLUSION: Immunohistochemical staining showed that TGF-β1 was mainly expressed in the cytoplasm. Seven days after transfection, the staining of TGF-β1 was positive on osteocytes of bone edge, osteocytes of woven bone callus and osteoblasts on the surface of bone callus. At 14 days, the expression of TGF-β1 was found on osteocytes of newborn woven bone callus, osteoblasts on the surface of bone callus, stromal cells in granulation tissue, mononuclear giant cells and multinucleated giant cells. At 28 days, the positive expression of TGF-β1 was decreased obviously. Compared with the injection of pIRES and normal saline, the expression of TGF-β1 was significantly higher after the injection of recombinant plasmid pIRES-hVEGF165-hBMP2, pIRES-hBMP2 and pIRES-hVEGF165 (P < 0.05 or P < 0.01). It suggests that gene therapy can promote TGF-β1 expression effectively and promote the formation of new bone and cell matrix in distraction gap.

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