中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2008年
7期
1372-1375
,共4页
背景:近年来,生长因子在肌腱愈合与粘连形成中的作用受到关注,其中转化生长因子与组织粘连与瘢痕形成的关系更倍受重视.目的:观察兔屈趾肌腱Ⅱ区伤口愈合过程中转化生长因子β1基因表达的变化.设计:随机对照动物实验.单位:青岛大学医学院附属医院骨科.材料:选用清洁级成年新西兰大白兔60只,体质量4.0～4.5 kg,雌雄不拘,由青岛市实验动物中心提供.所有动物的左前肢作为实验侧,同一动物右前肢作为对照侧.按术后1,7,14,21,28和56 d 6个时间点进行观察,每个时间点10只.其中6只进行原位杂交实验,4只进行免疫组织化学染色.实验过程中对动物的处置均符合动物伦理学标准.方法:实验于2005-09/2006-07在青岛大学医学院附属医院动物实验中心完成.麻醉后将所有动物左前中趾Ⅱ区屈趾深肌腱切断并用使用标准Kessler缝合法修复,对照侧不进行干预.分别于术后1,7,14,21,28和56 d麻醉后处死动物,实验侧沿原切口切开皮肤,切取肌腱与腱鞘.对照侧采取相同措施.主要观察指标:将肌腱与腱鞘组织进行原位杂交和免疫组织化学染色,观察转化生长因子β1的表达情况.结果:纳入的60只动物全部进入结果分析.①原位杂交结果:实验侧肌腱损伤后1 d,转化生长因子β1 Mrna的表达明显升高,在肌腱损伤后的14～21 d,转化生长因子β1 Mrna的表达持续升高达到最高峰,28 d开始下降,56d时仍保持较高水平.修复部位周围的腱鞘组织转化生长因子β1 Mrna的表达水平更高,在相同时间点,腱鞘细胞内的转化生长因子β1 Mrna的表达均高于肌腱组织.对照侧肌腱组织和腱鞘内均存在着转化生长因子β1 Mrna的表达,但是水平较低.实验侧各时间点腱鞘与肌腱细胞转化生长因子β1 Mrna的表达明显高于对照组,差异有显著性意义(P ＜ 0.05).②免疫组织化学染色结果:实验侧动物转化生长因子β1蛋白信号的表达术后第1 天开始增加,14～21 d达到高峰,56 d仍保持较高的水平.对照侧动物存在转化生长因子β1蛋白信号的表达,但表达水平较低.结论:正常无损伤的肌腱和腱鞘细胞能产生转化生长因子β1,当肌腱损伤后,细胞因子被激活,增加的细胞因子主要由肌腱细胞与腱鞘细胞产生,与肌腱的内、外源性愈合机制是一致的.
揹景:近年來,生長因子在肌腱愈閤與粘連形成中的作用受到關註,其中轉化生長因子與組織粘連與瘢痕形成的關繫更倍受重視.目的:觀察兔屈趾肌腱Ⅱ區傷口愈閤過程中轉化生長因子β1基因錶達的變化.設計:隨機對照動物實驗.單位:青島大學醫學院附屬醫院骨科.材料:選用清潔級成年新西蘭大白兔60隻,體質量4.0～4.5 kg,雌雄不拘,由青島市實驗動物中心提供.所有動物的左前肢作為實驗側,同一動物右前肢作為對照側.按術後1,7,14,21,28和56 d 6箇時間點進行觀察,每箇時間點10隻.其中6隻進行原位雜交實驗,4隻進行免疫組織化學染色.實驗過程中對動物的處置均符閤動物倫理學標準.方法:實驗于2005-09/2006-07在青島大學醫學院附屬醫院動物實驗中心完成.痳醉後將所有動物左前中趾Ⅱ區屈趾深肌腱切斷併用使用標準Kessler縫閤法脩複,對照側不進行榦預.分彆于術後1,7,14,21,28和56 d痳醉後處死動物,實驗側沿原切口切開皮膚,切取肌腱與腱鞘.對照側採取相同措施.主要觀察指標:將肌腱與腱鞘組織進行原位雜交和免疫組織化學染色,觀察轉化生長因子β1的錶達情況.結果:納入的60隻動物全部進入結果分析.①原位雜交結果:實驗側肌腱損傷後1 d,轉化生長因子β1 Mrna的錶達明顯升高,在肌腱損傷後的14～21 d,轉化生長因子β1 Mrna的錶達持續升高達到最高峰,28 d開始下降,56d時仍保持較高水平.脩複部位週圍的腱鞘組織轉化生長因子β1 Mrna的錶達水平更高,在相同時間點,腱鞘細胞內的轉化生長因子β1 Mrna的錶達均高于肌腱組織.對照側肌腱組織和腱鞘內均存在著轉化生長因子β1 Mrna的錶達,但是水平較低.實驗側各時間點腱鞘與肌腱細胞轉化生長因子β1 Mrna的錶達明顯高于對照組,差異有顯著性意義(P ＜ 0.05).②免疫組織化學染色結果:實驗側動物轉化生長因子β1蛋白信號的錶達術後第1 天開始增加,14～21 d達到高峰,56 d仍保持較高的水平.對照側動物存在轉化生長因子β1蛋白信號的錶達,但錶達水平較低.結論:正常無損傷的肌腱和腱鞘細胞能產生轉化生長因子β1,噹肌腱損傷後,細胞因子被激活,增加的細胞因子主要由肌腱細胞與腱鞘細胞產生,與肌腱的內、外源性愈閤機製是一緻的.
배경:근년래,생장인자재기건유합여점련형성중적작용수도관주,기중전화생장인자여조직점련여반흔형성적관계경배수중시.목적:관찰토굴지기건Ⅱ구상구유합과정중전화생장인자β1기인표체적변화.설계:수궤대조동물실험.단위:청도대학의학원부속의원골과.재료:선용청길급성년신서란대백토60지,체질량4.0～4.5 kg,자웅불구,유청도시실험동물중심제공.소유동물적좌전지작위실험측,동일동물우전지작위대조측.안술후1,7,14,21,28화56 d 6개시간점진행관찰,매개시간점10지.기중6지진행원위잡교실험,4지진행면역조직화학염색.실험과정중대동물적처치균부합동물윤리학표준.방법:실험우2005-09/2006-07재청도대학의학원부속의원동물실험중심완성.마취후장소유동물좌전중지Ⅱ구굴지심기건절단병용사용표준Kessler봉합법수복,대조측불진행간예.분별우술후1,7,14,21,28화56 d마취후처사동물,실험측연원절구절개피부,절취기건여건초.대조측채취상동조시.주요관찰지표:장기건여건초조직진행원위잡교화면역조직화학염색,관찰전화생장인자β1적표체정황.결과:납입적60지동물전부진입결과분석.①원위잡교결과:실험측기건손상후1 d,전화생장인자β1 Mrna적표체명현승고,재기건손상후적14～21 d,전화생장인자β1 Mrna적표체지속승고체도최고봉,28 d개시하강,56d시잉보지교고수평.수복부위주위적건초조직전화생장인자β1 Mrna적표체수평경고,재상동시간점,건초세포내적전화생장인자β1 Mrna적표체균고우기건조직.대조측기건조직화건초내균존재착전화생장인자β1 Mrna적표체,단시수평교저.실험측각시간점건초여기건세포전화생장인자β1 Mrna적표체명현고우대조조,차이유현저성의의(P ＜ 0.05).②면역조직화학염색결과:실험측동물전화생장인자β1단백신호적표체술후제1 천개시증가,14～21 d체도고봉,56 d잉보지교고적수평.대조측동물존재전화생장인자β1단백신호적표체,단표체수평교저.결론:정상무손상적기건화건초세포능산생전화생장인자β1,당기건손상후,세포인자피격활,증가적세포인자주요유기건세포여건초세포산생,여기건적내、외원성유합궤제시일치적.
BACKGROUND: We have paid more attention on the effects of growth factors on tendon healing and adhesion formation, especially on the correlation of transforming growth factor with tissue adhesion and scar formation. OBJECTIVE: To investigate the expression of transforming growth factor beta-1 mRNA in the zone Ⅱ flexor tendon of wound-healing rabbit models. DESIGN: Randomized controlled animal study. SETTING: Department of Orthopaedics, Affiliated Hospital of Medical College, Qingdao University. MATERIALS: Sixty clean adult New Zealand white rabbits weighting 4.0-4.5 kg, of either sex, were provided by Qingdao Animal Experimental Center. Left forelimbs of each animal were as experimental side, and right forelimbs of each animal were as control. There were 6 time points, namely at days 1, 7, 14, 21, 28 and 56, 10 rabbits in each time point. Of the 10 rabbits, 6 rabbits received the in situ hybridization and 4 rabbits received the immunohistochemical staining. Animal intervention met the animal ethical standard. METHODS: Experiments were performed at the Animal Experimental Center of Hospital Affiliated to Medical College of Qingdao University from September 2005 to July 2006. After anesthesia, each rabbit underwent complete transection of the profundus middle flexor tendon in zone Ⅱ, and then the tendon was repaired by the Kessler method. Rabbits in the control group did not receive any intervention. Rabbits were anesthetized and killed 1, 7, 14, 21, 28 and 56 days after the surgery. Skin was incised along the original incision at the experimental sides to obtain tendons and tendon sheaths. The same measurements were performed in the control group. MAIN OUTCOME MEASURES: Tenocytes and tendon sheath cells were detected with the in situ hybridization and the immunohistochemical staining to observe the expression of transforming growth factor beta-1. RESULTS: Sixty rabbits were involved in the result analysis. ①The in situ hybridization results: Expression of transforming growth factor beta-1 mRNA was increased at day 1 after tendon injury in the experimental group, reached a peak at days 14-21 after tendon injury, reduced at day 28 and was still in a high level at day 56. Expression of transforming growth factor beta-1 mRNA was high in tendon sheath cells around the repaired region. At the same time point, the expression of transforming growth factor beta-1 mRNA was higher in tendon sheath cells than in tenocytes. Low expression of transforming growth factor beta-1 mRNA was found in tenocytes and tendon sheath cells in the control group. The expression of transforming growth factor beta-1 mRNA in tenocytes and tendon sheath cells was higher in the experimental group than in the control group at each time point (P ＜ 0.05). ②Immunohistochemical staining results: Expression of transforming growth factor beta-1 protein was elevated at day 1 after the surgery, reached the peak at days 14-21 and was still in a high level at day 56 in the experimental group. Low expression of transforming growth factor beta-1 protein was seen in the control group. CONCLUSION: The normal uninjured tenocytes and tendon sheath cells produce transforming growth factor beta-1. The cytokine is activated in the injured tendon. The increase of this cytokine in both tenocytes and tendon sheath fibroblasts are coincidence with both extrinsic and intrinsic mechanisms for tendon repair.
