中华烧伤杂志
中華燒傷雜誌
중화소상잡지
16
2010年
4期
304-308
,共5页
林樾%梁红伟%李云剑%燕辛%谭谦
林樾%樑紅偉%李雲劍%燕辛%譚謙
림월%량홍위%리운검%연신%담겸
血小板源性生长因子%基因%转染%肌腱%组织粘连
血小闆源性生長因子%基因%轉染%肌腱%組織粘連
혈소판원성생장인자%기인%전염%기건%조직점련
Platelet-derived growth factor%Genes%Transfection%Tendon%Tissue adhesion
目的 了解血小板源性生长因子BB(PDGF-BB)基因转染大鼠肌腱细胞对肌腱愈合及肌腱粘连的影响. 方法 将90只SD大鼠制成跟腱损伤模型,按随机数字表法分为3组,每组30只:实验组,肌腱断端注射20μL转染PDGF-BB基因的大鼠肌腱细胞(1×108个/mL);对照组,肌腱断端注射20μL未行转染的大鼠肌腱细胞(1×108个/mL);空白对照组,不做任何处理.6-0丝线行改良Kessler法缝合跟腱,管型石膏固定1周.通过基因测序及RT-PCR鉴定转染PDGF-BB基因的大鼠肌腱细胞.分别于术后3 d和1、2、4、8周取各组大鼠肌腱组织样本,行大体、组织学观察以及生物力学检测,对比各组肌腱粘连度、组织中Fb数量与胶原纤维含量、肌腱最大抗拉力及最大滑动距离、组织中PDGF-BB的浓度.对数据行t检验. 结果(1)转染的肌腱细胞经RT-PCR以及基因测序证实在体外稳定表达PDGF-BB mRNA.(2)各组大鼠术后3 d肌腱均出现较明显肿胀及炎性细胞浸润,实验组改变较其他组明显轻微;随后各组情况均逐渐好转.术后4、8周肌腱粘连度分级组间比较未见明显差异.(3)实验组Fb数在术后2、4、8周显著低于对照组和空白对照组(t值分别为2.94、4.26、5.76和4.00、3.83、6.12,P＜0.05或P＜0.01).(4)实验组术后4周胶原纤维含量为(43±6)%,较对照组[(55±8)%]与空白对照组[(61±8)%]显著下降(t值分别为2.94和4.41,P＜0.05或P＜0.01).(5)术后4、8周实验组肌腱最大滑动距离为(3.25±0.33)、(3.65±0.21)mm,显著高于对照组的(2.29±0.40)、(2.21±0.37)mm和空白对照组的(2.01±0.23)、(1.89±0.24)mm(t值分别为4.53、8.29和7.55、13.52,P值均小于0.01),但其肌腱最大抗拉力与另2组比较差异无统计学意义(t值分别为0.41、0.41和0.77、0.72,P值均大于0.05).(6)术后3 d和2、4周,实验组肌腱组织中PDGF-BB浓度为(12.95±1.36)、(8.32±0.94)、(9.10±1.06)ng/mL,均显著高于对照组的(1.13±0.21)、(2.07±0.48)、(3.85±0.39)ng/mL(t值分别为21.04、14.50、11.39,P值均小于0.01). 结论 转染PDGF-BB基因肌腱细胞有促进肌腱内源性愈合、减轻肌腱粘连的作用.
目的 瞭解血小闆源性生長因子BB(PDGF-BB)基因轉染大鼠肌腱細胞對肌腱愈閤及肌腱粘連的影響. 方法 將90隻SD大鼠製成跟腱損傷模型,按隨機數字錶法分為3組,每組30隻:實驗組,肌腱斷耑註射20μL轉染PDGF-BB基因的大鼠肌腱細胞(1×108箇/mL);對照組,肌腱斷耑註射20μL未行轉染的大鼠肌腱細胞(1×108箇/mL);空白對照組,不做任何處理.6-0絲線行改良Kessler法縫閤跟腱,管型石膏固定1週.通過基因測序及RT-PCR鑒定轉染PDGF-BB基因的大鼠肌腱細胞.分彆于術後3 d和1、2、4、8週取各組大鼠肌腱組織樣本,行大體、組織學觀察以及生物力學檢測,對比各組肌腱粘連度、組織中Fb數量與膠原纖維含量、肌腱最大抗拉力及最大滑動距離、組織中PDGF-BB的濃度.對數據行t檢驗. 結果(1)轉染的肌腱細胞經RT-PCR以及基因測序證實在體外穩定錶達PDGF-BB mRNA.(2)各組大鼠術後3 d肌腱均齣現較明顯腫脹及炎性細胞浸潤,實驗組改變較其他組明顯輕微;隨後各組情況均逐漸好轉.術後4、8週肌腱粘連度分級組間比較未見明顯差異.(3)實驗組Fb數在術後2、4、8週顯著低于對照組和空白對照組(t值分彆為2.94、4.26、5.76和4.00、3.83、6.12,P＜0.05或P＜0.01).(4)實驗組術後4週膠原纖維含量為(43±6)%,較對照組[(55±8)%]與空白對照組[(61±8)%]顯著下降(t值分彆為2.94和4.41,P＜0.05或P＜0.01).(5)術後4、8週實驗組肌腱最大滑動距離為(3.25±0.33)、(3.65±0.21)mm,顯著高于對照組的(2.29±0.40)、(2.21±0.37)mm和空白對照組的(2.01±0.23)、(1.89±0.24)mm(t值分彆為4.53、8.29和7.55、13.52,P值均小于0.01),但其肌腱最大抗拉力與另2組比較差異無統計學意義(t值分彆為0.41、0.41和0.77、0.72,P值均大于0.05).(6)術後3 d和2、4週,實驗組肌腱組織中PDGF-BB濃度為(12.95±1.36)、(8.32±0.94)、(9.10±1.06)ng/mL,均顯著高于對照組的(1.13±0.21)、(2.07±0.48)、(3.85±0.39)ng/mL(t值分彆為21.04、14.50、11.39,P值均小于0.01). 結論 轉染PDGF-BB基因肌腱細胞有促進肌腱內源性愈閤、減輕肌腱粘連的作用.
목적 료해혈소판원성생장인자BB(PDGF-BB)기인전염대서기건세포대기건유합급기건점련적영향. 방법 장90지SD대서제성근건손상모형,안수궤수자표법분위3조,매조30지:실험조,기건단단주사20μL전염PDGF-BB기인적대서기건세포(1×108개/mL);대조조,기건단단주사20μL미행전염적대서기건세포(1×108개/mL);공백대조조,불주임하처리.6-0사선행개량Kessler법봉합근건,관형석고고정1주.통과기인측서급RT-PCR감정전염PDGF-BB기인적대서기건세포.분별우술후3 d화1、2、4、8주취각조대서기건조직양본,행대체、조직학관찰이급생물역학검측,대비각조기건점련도、조직중Fb수량여효원섬유함량、기건최대항랍력급최대활동거리、조직중PDGF-BB적농도.대수거행t검험. 결과(1)전염적기건세포경RT-PCR이급기인측서증실재체외은정표체PDGF-BB mRNA.(2)각조대서술후3 d기건균출현교명현종창급염성세포침윤,실험조개변교기타조명현경미;수후각조정황균축점호전.술후4、8주기건점련도분급조간비교미견명현차이.(3)실험조Fb수재술후2、4、8주현저저우대조조화공백대조조(t치분별위2.94、4.26、5.76화4.00、3.83、6.12,P＜0.05혹P＜0.01).(4)실험조술후4주효원섬유함량위(43±6)%,교대조조[(55±8)%]여공백대조조[(61±8)%]현저하강(t치분별위2.94화4.41,P＜0.05혹P＜0.01).(5)술후4、8주실험조기건최대활동거리위(3.25±0.33)、(3.65±0.21)mm,현저고우대조조적(2.29±0.40)、(2.21±0.37)mm화공백대조조적(2.01±0.23)、(1.89±0.24)mm(t치분별위4.53、8.29화7.55、13.52,P치균소우0.01),단기기건최대항랍력여령2조비교차이무통계학의의(t치분별위0.41、0.41화0.77、0.72,P치균대우0.05).(6)술후3 d화2、4주,실험조기건조직중PDGF-BB농도위(12.95±1.36)、(8.32±0.94)、(9.10±1.06)ng/mL,균현저고우대조조적(1.13±0.21)、(2.07±0.48)、(3.85±0.39)ng/mL(t치분별위21.04、14.50、11.39,P치균소우0.01). 결론 전염PDGF-BB기인기건세포유촉진기건내원성유합、감경기건점련적작용.
Objective To study the effect of platelet-derived growth factor-BB (PDGF-BB) gene transfected rat tendon cells on the healing and adhesion of rat tendon. Methods A model of heel tendon injury was reproduced in 90 rats. They were randomly divided into three groups: experiment group [with injection of 20 μL rat tendon cells(1 × 108 cell/mL) transfected with PDGF-BB gene into the injured tendon ends], control group [with injection of 20 μL non-transfected rat tendon cells (1 × 108 cell/mL) into the injured tendon ends] , and blank control group(without treatment) , with 30 rats in each group. Heel tendon ends were sutured with 6-0 thread by modified Kessler method and immobilized with tube-type plaster of Paris cast for one week. Rat tendon cells transfected with PDGF-BB gene were identified with gene sequencing and RT-PCR. Tendon tissue sample was harvested 3 days or 1, 2, 4, 8 week(s) after operation (POD or POW) for morphology and histology observation, and bio-mechanical test. The degree of tendon adhesion,the number of Fb and collagen fiber content in tissue, maximum tensile strength and sliding distance of tendon, and concentration of PDGF-BB in tendon tissue among groups were compared. Data were processed with t test. Results (1) PDGF-BB mRNA expressed stably in PDGF-BB gene transfected tendon cells as testified by RT-PCR and gene sequencing. (2) Obvious edema and inflammatory cells infiltration were observed in each group on POD 3, but they were less pronounced in experiment group. And the changes in all groups were ameliorated gradually. The difference in grading of tendon adhesion was not obvious among groups in POW 4 and 8. (3) Fb number in experiment group in POW 2, 4, 8 was respectively fewer than that of control group and blank control group(with t value respectively 2.94, 4. 26, 5.76 and 4.00, 3.83,6.12, P ＜0.05 orP ＜0.01). (4) Collagen fiber content in rat tendon of experimental group in POW 4was (43 ± 6) %, which was significantly lower as compared with that of control group [(55 ± 8) %] and blank control group [(61 ± 8) %](with t value respectively 2.94 and 4.41, P ＜ 0. 05 or P ＜ 0. 01). (5)The largest sliding distance of tendon in experiment group in POW 4 and 8 were (3.25 ±0.33) and (3.65 ±0.21) mm, which were significantly longer than those in control group [(2.29 ±0.40), (2.21 ±0.37)mm] and blank control group [(2.01 ± 0.23),(1.89 ± 0. 24) mm](with t value respectively 4.53, 8.29and 7.55, 13.52, P values all below 0.01). There was no statistical significant difference among the three groups in the maximum tensile strength of tendon (with t value respectively 0. 41,0.41,0.77, 0.72, P values all above 0. 05). (6) Content of PDGF-BB in tendon tissue of experimental group on POD 3 and in POW 2, 4 were(12.95 ± 1.36),(8.32 ± 0. 94), (9.10 ± 1.06) ng/mL, all significantly higher than those in control group [(1.13 ±0.21),(2.07 ± 0.48),(3.85 ± 0.39) ng/mL](with t value respectively 21.04,14. 50, 11.39, P values all below 0. 01). Conclusions PDGF-BB gene transfected rat tendon cells can promote endogenous healing of tendon and prevent tendon adhesion.
