Wang Xiao Tian, Liu Paul Y, Tang Jin Bo
Department of Surgery, Roger Williams Medical Center, Boston University School of Medicine, Providence, RI 02908-4735, USA.
J Hand Surg Am. 2004 Sep;29(5):884-90. doi: 10.1016/j.jhsa.2004.05.016.
Promotion of collagen production can increase tendon healing strength and reduce repair ruptures. Transfer of an exogenous growth factor gene to tenocytes of intrasynovial tendons may enhance the capacity of cells to produce collagen. We transferred the platelet-derived growth factor B (PDGF-B) gene to tenocytes and investigated its effects on the expression of the PDGF gene and the type I collagen gene in an in vitro tenocyte culture model.
Tenocytes obtained from explant cultures of rat intrasynovial tendons were treated for 12 hours with the plasmid containing the PDGF complementary deoxyribonucleic acid (cDNA) with liposome and were then cultured for 6 additional days. The control tenocytes did not receive the exogenous gene and liposome. Efficiency of the gene transfer was evaluated by using reverse transcription polymerase chain reactions (RT-PCR) to detect the presence of the transferred gene in the tenocytes. Enhancement of the expression of the target gene was assessed by RT-PCR with primers effective to amplify both internal and transferred genes. Expression of the type I collagen gene was determined by quantitative analysis of the products of RT-PCR.
Levels of expression of the type I collagen gene by tenocytes were increased significantly by transfer of the exogenous PDGF gene to the tenocytes. Efficiency of the gene transfer was confirmed by the presence of exogenous PDGF cDNA in the tenocytes receiving the transferred gene. Expression of the PDGF gene increased significantly in the cells treated with exogenous PDGF cDNA.
Exogenous PDGF genes can be transferred effectively into intrasynovial tenocytes and the transfer increases significantly the expression of genes for PDGF and type I collagen. Transfer of the PDGF gene may offer a novel way of effectively promoting healing of intrasynovial flexor tendons. The findings warrant future in vivo study to test the effectiveness of gene therapy to promote flexor tendon healing.
促进胶原蛋白生成可增强肌腱愈合强度并减少修复破裂。将外源性生长因子基因转移至滑膜内肌腱的腱细胞可能会增强细胞产生胶原蛋白的能力。我们将血小板衍生生长因子B（PDGF - B）基因转移至腱细胞，并在体外腱细胞培养模型中研究其对PDGF基因和I型胶原蛋白基因表达的影响。
从大鼠滑膜内肌腱的外植体培养物中获取的腱细胞，用含有PDGF互补脱氧核糖核酸（cDNA）的质粒与脂质体处理12小时，然后再培养6天。对照腱细胞未接受外源性基因和脂质体。通过逆转录聚合酶链反应（RT - PCR）检测腱细胞中转移基因的存在来评估基因转移效率。使用对扩增内部和转移基因均有效的引物，通过RT - PCR评估靶基因表达的增强情况。通过对RT - PCR产物的定量分析来确定I型胶原蛋白基因的表达。
将外源性PDGF基因转移至腱细胞后，腱细胞中I型胶原蛋白基因的表达水平显著增加。接受转移基因的腱细胞中存在外源性PDGF cDNA，证实了基因转移效率。用外源性PDGF cDNA处理的细胞中PDGF基因的表达显著增加。
外源性PDGF基因可有效转移至滑膜内腱细胞，且这种转移显著增加了PDGF和I型胶原蛋白基因的表达。PDGF基因的转移可能为有效促进滑膜内屈肌腱愈合提供一种新方法。这些发现值得未来进行体内研究以测试基因治疗促进屈肌腱愈合的有效性。
