CAJ | 학술논문

背景:研究表明,转化生长因子β1在肌腱损伤愈合过程中增加了肌腱细胞胶原的合成和术后粘连的形成.目的:观察转化生长因子β1抗体对转化生长因子β诱导的肌腱细胞胶原产生及术后粘连形成的影响.设计、时间及地点:随机分组观察实验,于2005-09/2006-06在同济医学院实验动物中心完成.材料:选择2-5月龄新西兰大白兔,体质量3.5～4.5 kg.转化生长因子由美国SantaCruz B iotechnology公司提供.方法:取兔屈指肌腱分离肌腱成纤维细胞、腱外膜细胞和腱内膜细胞,将细胞随机分成2组,实验组加入1 μg/L转化生长因子β后,再加入0.1,0.5,1.0mg/L转化生长因子β1中和抗体,对照组不添加任何试剂,酶联免疫吸附实验测定Ⅰ型胶原.取84只兔行中趾屈指肌腱切断吻合术,将其中36只兔随机分成3组,腱鞘内分别注入生理盐水、1.0,2.0 mg/L转化生长因子β1中和抗体,4,8周后取出肌腱行肌腱粘连检测、生物力学测定、组织学观察和扫描电镜观察;余48只兔随机分成2组,腱鞘内分别注入生理盐水和1.0mg/L转化生长因子β1中和抗体,1,2,4,8周后取出肌腱,原位杂交方法测定肌腱转化生长因子β1和Ⅰ型胶原mRNA的表达.主要观察指标:各组兔肌腱细胞胶原产生及术后粘连情况.结果:酶联免疫吸附实验显示,转化生长因子β1能明显提高肌腱细胞Ⅰ型胶原的产生;转化生长因子β1抗体能降低3种细胞Ⅰ型胶原的产生,随抗体质量浓度增加,Ⅰ型胶原水平逐渐降低,且呈剂量依赖性;术后4,8周,与1.0,2.0mg/L转化生长因子β1组比较,生理盐水组屈趾肌腱滑动距离较短,模拟主动屈曲度明显受限(P<0.05),最大抗断裂载荷各组间比较差异无显著性意义(P>0.05).扫描电镜和组织学观察结果显示,术后4,8周生理盐水组胶原纤维排列紊乱,1.0,2.0 mg/L转化生长因子β1组胶原纤维排列整齐.原位杂交结果显示,术后各时间点1.0 mg/L转化生长因子β1组转化生长因子β1和Ⅰ型胶原mRNA表达均低于生理盐水组(P<0.05).结论:转化生长因子β1抗体能有效抑制转化生长因子β1在肌腱损伤修复中的作用,减少粘连形成. 揹景:研究錶明,轉化生長因子β1在肌腱損傷愈閤過程中增加瞭肌腱細胞膠原的閤成和術後粘連的形成.目的:觀察轉化生長因子β1抗體對轉化生長因子β誘導的肌腱細胞膠原產生及術後粘連形成的影響.設計、時間及地點:隨機分組觀察實驗,于2005-09/2006-06在同濟醫學院實驗動物中心完成.材料:選擇2-5月齡新西蘭大白兔,體質量3.5～4.5 kg.轉化生長因子由美國SantaCruz B iotechnology公司提供.方法:取兔屈指肌腱分離肌腱成纖維細胞、腱外膜細胞和腱內膜細胞,將細胞隨機分成2組,實驗組加入1 μg/L轉化生長因子β後,再加入0.1,0.5,1.0mg/L轉化生長因子β1中和抗體,對照組不添加任何試劑,酶聯免疫吸附實驗測定Ⅰ型膠原.取84隻兔行中趾屈指肌腱切斷吻閤術,將其中36隻兔隨機分成3組,腱鞘內分彆註入生理鹽水、1.0,2.0 mg/L轉化生長因子β1中和抗體,4,8週後取齣肌腱行肌腱粘連檢測、生物力學測定、組織學觀察和掃描電鏡觀察;餘48隻兔隨機分成2組,腱鞘內分彆註入生理鹽水和1.0mg/L轉化生長因子β1中和抗體,1,2,4,8週後取齣肌腱,原位雜交方法測定肌腱轉化生長因子β1和Ⅰ型膠原mRNA的錶達.主要觀察指標:各組兔肌腱細胞膠原產生及術後粘連情況.結果:酶聯免疫吸附實驗顯示,轉化生長因子β1能明顯提高肌腱細胞Ⅰ型膠原的產生;轉化生長因子β1抗體能降低3種細胞Ⅰ型膠原的產生,隨抗體質量濃度增加,Ⅰ型膠原水平逐漸降低,且呈劑量依賴性;術後4,8週,與1.0,2.0mg/L轉化生長因子β1組比較,生理鹽水組屈趾肌腱滑動距離較短,模擬主動屈麯度明顯受限(P<0.05),最大抗斷裂載荷各組間比較差異無顯著性意義(P>0.05).掃描電鏡和組織學觀察結果顯示,術後4,8週生理鹽水組膠原纖維排列紊亂,1.0,2.0 mg/L轉化生長因子β1組膠原纖維排列整齊.原位雜交結果顯示,術後各時間點1.0 mg/L轉化生長因子β1組轉化生長因子β1和Ⅰ型膠原mRNA錶達均低于生理鹽水組(P<0.05).結論:轉化生長因子β1抗體能有效抑製轉化生長因子β1在肌腱損傷脩複中的作用,減少粘連形成.
배경:연구표명,전화생장인자β1재기건손상유합과정중증가료기건세포효원적합성화술후점련적형성.목적:관찰전화생장인자β1항체대전화생장인자β유도적기건세포효원산생급술후점련형성적영향.설계、시간급지점:수궤분조관찰실험,우2005-09/2006-06재동제의학원실험동물중심완성.재료:선택2-5월령신서란대백토,체질량3.5～4.5 kg.전화생장인자유미국SantaCruz B iotechnology공사제공.방법:취토굴지기건분리기건성섬유세포、건외막세포화건내막세포,장세포수궤분성2조,실험조가입1 μg/L전화생장인자β후,재가입0.1,0.5,1.0mg/L전화생장인자β1중화항체,대조조불첨가임하시제,매련면역흡부실험측정Ⅰ형효원.취84지토행중지굴지기건절단문합술,장기중36지토수궤분성3조,건초내분별주입생리염수、1.0,2.0 mg/L전화생장인자β1중화항체,4,8주후취출기건행기건점련검측、생물역학측정、조직학관찰화소묘전경관찰;여48지토수궤분성2조,건초내분별주입생리염수화1.0mg/L전화생장인자β1중화항체,1,2,4,8주후취출기건,원위잡교방법측정기건전화생장인자β1화Ⅰ형효원mRNA적표체.주요관찰지표:각조토기건세포효원산생급술후점련정황.결과:매련면역흡부실험현시,전화생장인자β1능명현제고기건세포Ⅰ형효원적산생;전화생장인자β1항체능강저3충세포Ⅰ형효원적산생,수항체질량농도증가,Ⅰ형효원수평축점강저,차정제량의뢰성;술후4,8주,여1.0,2.0mg/L전화생장인자β1조비교,생리염수조굴지기건활동거리교단,모의주동굴곡도명현수한(P<0.05),최대항단렬재하각조간비교차이무현저성의의(P>0.05).소묘전경화조직학관찰결과현시,술후4,8주생리염수조효원섬유배렬문란,1.0,2.0 mg/L전화생장인자β1조효원섬유배렬정제.원위잡교결과현시,술후각시간점1.0 mg/L전화생장인자β1조전화생장인자β1화Ⅰ형효원mRNA표체균저우생리염수조(P<0.05).결론:전화생장인자β1항체능유효억제전화생장인자β1재기건손상수복중적작용,감소점련형성.
BACKGROUND: Studies have showed that transforming growth factor-β1 (TGF-β1) could yield to the collagen synthesis and adhesion formation of tendon cells at the process of healing. OBJECTIVE: To investigate the preventive effect of TGF-β1 neutralizing antibody on the collagen production and adhesion formation of flexor tendon. DESIGN, TIME AND SETTING: Randomized grouping observational experiments were performed in the Experimental Animal Center of Tongji Medical College between September 2005 and June 2006. MATERIALS: New Zealand white rabbits aged 2-5 months, weighing 3.5-4.5 kg. TGF was offered by Santa Cruz Biotechnology, USA. METHODS: Sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes were obtained from rabbit flexor tendons. Cells were divided into two groups at random. In the experiment group, each cell culture was supplemented with 1 μg/L of TGF-β at increasing dose (0.1, 0.5, 1.0 mg/L) of TGF-β1 neutralizing antibody. No reagents were given in the control group. Collagen Ⅰ production was measured by enzyme-linked immunoabsorbent assay. Eighty-four adult New Zealand white rabbit forepaws underwent sharp transection of middle toe flexor digitorum profundus, followed by immediate repair. Thirty-six adult New Zealand white rabbit were divided into three groups randomly (n=12), injecting with the saline, 1.0 mg/L TGF-β1 neutralizing antibody and 2.0 mg/L TGF-β1 neutralizing antibody into tendon sheath respectively. Tendons were harvested at 4 and 8 weeks to conduct adhesion detection, biomechanical testing, histological evaluation and scanning electron microscopy observation. The remaining 48 New Zealand white rabbits were divided into two groups randomly (n=24), undergoing the saline and 1,0 mg/L TGF-β1 neutralizing antibody injection in tendon sheath respectively. Tendons were harvested at an increasing time interval (1, 2, 4, 8 weeks) and analyzed by in situ hybridization to determine the mRNA expression of TGF-β1 and collagen Ⅰ. MAIN OUTCOME MEASURES: Collagen production and adhesion of rabbit tendon cells. RESULTS: ELISA exhibited that TGF-β1 increased collagen Ⅰ production and the addition of neutralizing antibody significantly reduced TGF-β-induced collagen Ⅰ production in all cell cultures. The effect between antibody and collagen Ⅰ was dose dependent. At 4 and 8 weeks after operation, the gliding excursion ratio of the tendon was shortened and the simulated active flexion ratio were less in saline group compared with 1.0 and 2.0 mg/L TGF-β1 groups (P < 0.05). The tendon anastomosis breaking strength was shown no significant differences among 3 groups (P > 0.05). Scanning electron microscopy and histological observation showed that collagen fibers arranged irregularly in saline group, but arranged regularly in 1.0 and 2.0 mg/L TGF-β1 groups at 4 and 8 weeks after operation. The in situ hybridization examination revealed that TGF-β1 and collagen Ⅰ mRNA expression in 1.0 mg/L TGF-β1 group was lower than that in saline group at each time (P < 0.05). CONCLUSION: TGF-β1 neutralizing antibody can inhibit the function of the TGF-β1 effectively following the flexor tendon injury and repair, and can prevent adhesion formation.