中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2014年
19期
1494-1498
,共5页
周盛源%朱巍%陈雄生%许政%钱闯%刘晓东
週盛源%硃巍%陳雄生%許政%錢闖%劉曉東
주성원%주외%진웅생%허정%전틈%류효동
韧带%创伤和损伤%成纤维细胞生长因子,碱性%干细胞
韌帶%創傷和損傷%成纖維細胞生長因子,堿性%榦細胞
인대%창상화손상%성섬유세포생장인자,감성%간세포
Ligaments%Wound and injuries%Fibroblast growth factor,basic%Stem cells
目的 评价转染碱性成纤维细胞生长因子(bFGF)基因的骨髓间充质干细胞(BMSCs)对严重韧带损伤的修复作用.方法 通过腺病毒将bFGF基因转入BMSCs,并标记增强型绿色荧光蛋白(eGFP).建立新西兰大白兔髌韧带损伤模型,在断裂韧带缝合后,根据局部细胞移植情况将手术组分为转基因BMSCs组(组1)、BMSCs组(组2)、无细胞组(组3)、正常韧带组(组4).术后不同时间点观察韧带损伤修复区转基因BMSCs的分布,bFGF表达及韧带特异性细胞外基质:Ⅰ型胶原、Ⅲ型胶原、fibronectin、α-SMA和vimentin的表达;组织学观察各组韧带损伤愈合情况;生物力学检测评价各组的生物力学强度.结果 转基因BMSCs组术后1个月内在韧带修复区内可持续观察到转基因BMSCs的存在,组1术后3～14 d bFGF表达明显高于对照组(P ＜0.001),其中移植后7d为表达高峰.术后1个月内转基因BMSCs组各韧带特异性细胞外基质基因表达明显强于对照组(P＜0.001),免疫组化呈强阳性表现.术后3个月HE染色见转基因BMSCs组韧带损伤修复区胶原纤维排列及致密度均优于对照组,生物力学检测表明转基因BMSCs组拉力载荷明显强于对照组[组1～4分别为(13.5±1.3)、(8.6±2.0)、(8.4±2.7)、(20.5±4.9)N,P＜0.05].结论 转染bFGF基因的BMSCs能明显促进韧带损伤愈合,增加韧带愈合强度.
目的 評價轉染堿性成纖維細胞生長因子(bFGF)基因的骨髓間充質榦細胞(BMSCs)對嚴重韌帶損傷的脩複作用.方法 通過腺病毒將bFGF基因轉入BMSCs,併標記增彊型綠色熒光蛋白(eGFP).建立新西蘭大白兔髕韌帶損傷模型,在斷裂韌帶縫閤後,根據跼部細胞移植情況將手術組分為轉基因BMSCs組(組1)、BMSCs組(組2)、無細胞組(組3)、正常韌帶組(組4).術後不同時間點觀察韌帶損傷脩複區轉基因BMSCs的分佈,bFGF錶達及韌帶特異性細胞外基質:Ⅰ型膠原、Ⅲ型膠原、fibronectin、α-SMA和vimentin的錶達;組織學觀察各組韌帶損傷愈閤情況;生物力學檢測評價各組的生物力學彊度.結果 轉基因BMSCs組術後1箇月內在韌帶脩複區內可持續觀察到轉基因BMSCs的存在,組1術後3～14 d bFGF錶達明顯高于對照組(P ＜0.001),其中移植後7d為錶達高峰.術後1箇月內轉基因BMSCs組各韌帶特異性細胞外基質基因錶達明顯彊于對照組(P＜0.001),免疫組化呈彊暘性錶現.術後3箇月HE染色見轉基因BMSCs組韌帶損傷脩複區膠原纖維排列及緻密度均優于對照組,生物力學檢測錶明轉基因BMSCs組拉力載荷明顯彊于對照組[組1～4分彆為(13.5±1.3)、(8.6±2.0)、(8.4±2.7)、(20.5±4.9)N,P＜0.05].結論 轉染bFGF基因的BMSCs能明顯促進韌帶損傷愈閤,增加韌帶愈閤彊度.
목적 평개전염감성성섬유세포생장인자(bFGF)기인적골수간충질간세포(BMSCs)대엄중인대손상적수복작용.방법 통과선병독장bFGF기인전입BMSCs,병표기증강형록색형광단백(eGFP).건립신서란대백토빈인대손상모형,재단렬인대봉합후,근거국부세포이식정황장수술조분위전기인BMSCs조(조1)、BMSCs조(조2)、무세포조(조3)、정상인대조(조4).술후불동시간점관찰인대손상수복구전기인BMSCs적분포,bFGF표체급인대특이성세포외기질:Ⅰ형효원、Ⅲ형효원、fibronectin、α-SMA화vimentin적표체;조직학관찰각조인대손상유합정황;생물역학검측평개각조적생물역학강도.결과 전기인BMSCs조술후1개월내재인대수복구내가지속관찰도전기인BMSCs적존재,조1술후3～14 d bFGF표체명현고우대조조(P ＜0.001),기중이식후7d위표체고봉.술후1개월내전기인BMSCs조각인대특이성세포외기질기인표체명현강우대조조(P＜0.001),면역조화정강양성표현.술후3개월HE염색견전기인BMSCs조인대손상수복구효원섬유배렬급치밀도균우우대조조,생물역학검측표명전기인BMSCs조랍력재하명현강우대조조[조1～4분별위(13.5±1.3)、(8.6±2.0)、(8.4±2.7)、(20.5±4.9)N,P＜0.05].결론 전염bFGF기인적BMSCs능명현촉진인대손상유합,증가인대유합강도.
Objective To evaluate the efficacies of transgenetic bone mesenchymal stem cells (BMSCs) carring basic fibroblast growth factor (bFGF) gene on repairing severe ligament injury.Methods The genes of bFGF and enhanced green fluorescent protein (eGFP) were transfected into BMSCs through adenovirus vector.According to different cell transplantations,the operated rabbits were divided into 3groups of transgenetic BMSCs,BMSCs and no cells.And group 4 was normal ligament group.The distribution of transgenetic BMSCs in ligament injury area,the expression of bFGF and ligament specific extracellular matrix,i.e.type Ⅰ collagen,type Ⅲ collagen,fibronectin,alpha-smooth muscle actin and vimentin,histology and biomechanics of ligament healing were recorded at different timepoints.Results The transgenetic BMSCs could be observed in injury area within one month.The expression of bFGF in the transgenetic BMSCs group was significantly higher than that in the control groups at Days 3-14 (P ＜0.001) and peaked at Day 7.In the transgenetic BMSCs group,the gene expression of ligament specific extracellular matrix was significantly higher than that in the control groups within one month (P ＜ 0.001).And immunohistochemistry showed strong positive expression.After 3 months,in group 1,hematoxylin and eosin staining showed the arrangement and density of collagen fibers of healing area were better than that in the control groups.And biomechanical testing showed that tensile load were stronger than that in the control groups (P ＜ 0.05).Conclusion Transgenetic BMSCs carrying bFGF gene can obviously promote ligament injury healing and increase ligament healing strength.
