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
28期
5589-5592
,共4页
背景:肌腱组织细胞具有分泌胶原的功能,在伤口愈合及粘连中有重要的作用.知之较少的是乳酸对肌腱细胞的生物学作用.目的:探讨兔屈趾肌腱腱鞘、腱外膜和腱内膜细胞增殖、胶原产生和乳酸对细胞的增殖、胶原产生和对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8分泌的影响.设计、时间和地点:随机对照动物实验,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.材料:选用6只成年清洁级新西兰大白兔,雌雄不拘.方法:①进行兔腱鞘成纤维细胞、腱外膜和腱内膜成纤维样细胞的分离和培养,亚甲蓝染色观察细胞形态并鉴定3种细胞类型.②以加入25 mmol/L乳酸培养细胞,测量细胞的数量.③比较加入25,50,100和200 mmol/L乳酸后细胞胶原产生量和对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8分泌量,并与不加乳酸培养为对照比较.主要观察指标:使用免疫组化检测Ⅰ、Ⅱ、Ⅲ型胶原的产生,通过酶联免疫吸附试验定量检测不同剂量乳酸对细胞Ⅰ型胶原产生的影响及对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8产生的影响.结果;①25 mmol/L乳酸使培养的细胞数量降低,3种细胞组间比较差异无显著性意义OP>0.05).②乳酸可以使实验组Ⅰ、Ⅱ和Ⅲ型胶原组织产生明显多于对照组,差异有显著性意义(P<0.05).③当乳酸浓度增加至50 mmol/L时,实验组3种细胞的Ⅰ型胶原量增加达最高峰,与乳酸浓度为0 mmol/L时比较,差异有非常显著性意义(t=4.58,3.97,3.16,P<0.01);当乳酸浓度增加到100 mmol/L和200 mmol/L时,Ⅰ型胶原产生量明显减少.④实验组乳酸浓度为25 mmol/L时对转化生长因子β1,基础纤维母细胞生长因子,白细胞介素8的分泌量增加,白细胞介素8的分泌量减少,与对照组比较差异均有显著性意义(P<0.05).结论:乳酸能增加腱鞘成纤维细胞、腱外膜细胞和腱内膜细胞的胶原产生量,增加的程度与乳酸的浓度有关,以50 mmol/L时最佳,而这种刺激作用可能与增加转化生长因子β1,基础纤维母细胞生长因子和减少白细胞介素8的分泌量有关.
揹景:肌腱組織細胞具有分泌膠原的功能,在傷口愈閤及粘連中有重要的作用.知之較少的是乳痠對肌腱細胞的生物學作用.目的:探討兔屈趾肌腱腱鞘、腱外膜和腱內膜細胞增殖、膠原產生和乳痠對細胞的增殖、膠原產生和對轉化生長因子β1,基礎纖維母細胞生長因子,白細胞介素8分泌的影響.設計、時間和地點:隨機對照動物實驗,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.材料:選用6隻成年清潔級新西蘭大白兔,雌雄不拘.方法:①進行兔腱鞘成纖維細胞、腱外膜和腱內膜成纖維樣細胞的分離和培養,亞甲藍染色觀察細胞形態併鑒定3種細胞類型.②以加入25 mmol/L乳痠培養細胞,測量細胞的數量.③比較加入25,50,100和200 mmol/L乳痠後細胞膠原產生量和對轉化生長因子β1,基礎纖維母細胞生長因子,白細胞介素8分泌量,併與不加乳痠培養為對照比較.主要觀察指標:使用免疫組化檢測Ⅰ、Ⅱ、Ⅲ型膠原的產生,通過酶聯免疫吸附試驗定量檢測不同劑量乳痠對細胞Ⅰ型膠原產生的影響及對轉化生長因子β1,基礎纖維母細胞生長因子,白細胞介素8產生的影響.結果;①25 mmol/L乳痠使培養的細胞數量降低,3種細胞組間比較差異無顯著性意義OP>0.05).②乳痠可以使實驗組Ⅰ、Ⅱ和Ⅲ型膠原組織產生明顯多于對照組,差異有顯著性意義(P<0.05).③噹乳痠濃度增加至50 mmol/L時,實驗組3種細胞的Ⅰ型膠原量增加達最高峰,與乳痠濃度為0 mmol/L時比較,差異有非常顯著性意義(t=4.58,3.97,3.16,P<0.01);噹乳痠濃度增加到100 mmol/L和200 mmol/L時,Ⅰ型膠原產生量明顯減少.④實驗組乳痠濃度為25 mmol/L時對轉化生長因子β1,基礎纖維母細胞生長因子,白細胞介素8的分泌量增加,白細胞介素8的分泌量減少,與對照組比較差異均有顯著性意義(P<0.05).結論:乳痠能增加腱鞘成纖維細胞、腱外膜細胞和腱內膜細胞的膠原產生量,增加的程度與乳痠的濃度有關,以50 mmol/L時最佳,而這種刺激作用可能與增加轉化生長因子β1,基礎纖維母細胞生長因子和減少白細胞介素8的分泌量有關.
배경:기건조직세포구유분비효원적공능,재상구유합급점련중유중요적작용.지지교소적시유산대기건세포적생물학작용.목적:탐토토굴지기건건초、건외막화건내막세포증식、효원산생화유산대세포적증식、효원산생화대전화생장인자β1,기출섬유모세포생장인자,백세포개소8분비적영향.설계、시간화지점:수궤대조동물실험,우2005-09/2006-07재청도대학의학원부속의원동물실험중심완성.재료:선용6지성년청길급신서란대백토,자웅불구.방법:①진행토건초성섬유세포、건외막화건내막성섬유양세포적분리화배양,아갑람염색관찰세포형태병감정3충세포류형.②이가입25 mmol/L유산배양세포,측량세포적수량.③비교가입25,50,100화200 mmol/L유산후세포효원산생량화대전화생장인자β1,기출섬유모세포생장인자,백세포개소8분비량,병여불가유산배양위대조비교.주요관찰지표:사용면역조화검측Ⅰ、Ⅱ、Ⅲ형효원적산생,통과매련면역흡부시험정량검측불동제량유산대세포Ⅰ형효원산생적영향급대전화생장인자β1,기출섬유모세포생장인자,백세포개소8산생적영향.결과;①25 mmol/L유산사배양적세포수량강저,3충세포조간비교차이무현저성의의OP>0.05).②유산가이사실험조Ⅰ、Ⅱ화Ⅲ형효원조직산생명현다우대조조,차이유현저성의의(P<0.05).③당유산농도증가지50 mmol/L시,실험조3충세포적Ⅰ형효원량증가체최고봉,여유산농도위0 mmol/L시비교,차이유비상현저성의의(t=4.58,3.97,3.16,P<0.01);당유산농도증가도100 mmol/L화200 mmol/L시,Ⅰ형효원산생량명현감소.④실험조유산농도위25 mmol/L시대전화생장인자β1,기출섬유모세포생장인자,백세포개소8적분비량증가,백세포개소8적분비량감소,여대조조비교차이균유현저성의의(P<0.05).결론:유산능증가건초성섬유세포、건외막세포화건내막세포적효원산생량,증가적정도여유산적농도유관,이50 mmol/L시최가,이저충자격작용가능여증가전화생장인자β1,기출섬유모세포생장인자화감소백세포개소8적분비량유관.
BACKGROUND: Tendon cells possess collagen-secreting function, which plays an important role in the wound healing and adhesion. Little is known about the biological effects of lactate on tendon cells. OBJECTIVE: This study was designed to study the proliferation and collagen production of tendon sheath fibroblasts,epitenon tenocytes, and endotenon tenocytes and investigate the effect of lactate on cell proliferation, collage production and secretion of transforming growth factor- β1(TGF- β 1),basic fibroblast growth factor (bFGF), and interleukin-8 (IL-8) by each of these 3 cell types in rabbit flexor tendon.DESIGN, TIME AND SETTING: This study, a randomized controlled animal experiment, was performed at the Animal Laboratory Center, Affiliated Hospital of Qingdao University Medical College between September 2005 and July 2006.MATERIALS: Six adult New Zealand clean rabbits of either gender were included in the present study.METHODS: Tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes were isolated from rabbit flexor tendon and cultured. Each cell type was identified by observing morphological structure through methylene blue staining.Three types of cells were cultured with media supplemented with 25 mol/L lactate to obtain cell number. The collagen production and secretion of TGF- β 1, b-FGF and FL-8 were compared after adding culture media supplemented with 25, 50,100, and 200 mmol/L lactate, respectively. At the same time, the above-mentioned indices measured after adding medium were compared with those measured without adding lactate.MAIN OUTCOME MEASURES: Type Ⅰ, Ⅱ, and Ⅲ collagen production was detected by immunohistochemistry; The effects of different concentrations of lactate on type Ⅰ collagen production as well as secretion of TGF- β 1,b-FGF, and IL-8by an enzyme linked immunosorbent assay (ELISA).RESULTS: 25 mmol/L lactate reduced 3 types of cultured cells. There was no significant difference in the cell number among the 3 types of cells (P > 0.05) Lactate produced more type Ⅰ, Ⅱ ,and Ⅲ collagen tissue compared to not adding lactate (P < 0.05) When lactate concentration increased to 50 mmol/L, type Ⅰ collagen reached its peak level in the 3 types of cells. There was significant difference in type Ⅰ collagen compared with lactate concentration was 0 mmol/L(t = 4.58, 3.97,3.16, P < 0.01 ) . When lactate concentration increased to 100 mmol/L and 200 mmol/L, type Ⅰ collagen was noticeably decreased. When lactate concentration was 25 mmol/L in the 3 types of cells, the secretion of TGF- β 1 and bFGF was increased and the secretion of IL-8 was decreased. There was significant difference compared with not adding lactate (P <0.05).CONCLUSION: Lactate can increase the collagen production of tendon sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes. The increasing degree of collagen production is related to lactate concentration, in particular at 50 mmol/L. Such a stimulation may be related to the increase of TGF- β 1 and bFGF and the decrease of IL-8 secretion.
