>><< 10 g/l 10 g/l 2 0 DNAIB DNA B BDNAI / d 10 TBDSCIENCE SIGMA SIGMASIGMA Hanks 10 min Hanks10 ml 10 ml 160 r/min40

Size: px

Start display at page:

Download ">><< 10 g/l 10 g/l 2 0 DNAIB DNA B BDNAI / d 10 TBDSCIENCE SIGMA SIGMASIGMA Hanks 10 min Hanks10 ml 10 ml 160 r/min40"

弁符蔺
7 years ago
Views:

1 Journal of Clinical Rehabilitative Tissue Engineering Research January 8, 2008 Vol.12, No Dose-effect relationship of insulin-like growth factor and the proliferation of tenocytes in tissue engineering Qiu Nan-hai 1, Xia Ying-peng 2, Li Ming-xin 3, Li Rui-hua 3, Kan Shi-lian 3 Abstract AIM: Insulin-like growth factor (IGF-) is a potent mitogen and powerful stimulator for the division and growth of tenocytes. In this study, we cultured the tenocytes of the second generation with IGF-at different doses to verify the effects of IGF-on cell proliferation and the dose effect. METHODS: The experiment was performed in the laboratory of Tianjin Hospital from November 2004 to April France Lohmann fertilized eggs were provided by the laboratory of Tianjin Hospital. On day 19, the tendons of 10 embryo chickens were obtained. Tenocytes were isolated and cultured. The cell morphous and growth rules were observed. The second generation cells were seeded onto 6-well culture plate and added with fetal calf serum (FCS) of different concentrations. The effects of serum concentration on cell attachment and growth were observed. The second generation tenocytes were seeded onto 96-well culture plate and divided into 7 groups. The former 5 groups were cultured in 0.02 volume fraction FCS culture medium containing IGF-at doses of 1, 5, 10, 50, 200g/L; the sixth group was cultured in 0.05 volume fraction FCS culture medium as positive control, and the seventh group was cultured in 0.02 volume fraction FCS culture medium as negative control. The effects of IGF-on cell proliferation were observed with MTT test and Swiss-gimmsa taint test. RESULTS: The tenocytes grew very fast after attaching to the wall. The primary cells started to passage in about 1 week. The proliferative rate of tenocytes was positively correlated with the concentration of FCS in culture medium. The proliferative rate of tenocytes was accelerated with the increase in IGF-dose. There was no difference among the 1g/L and 5g/L IGF-groups and negative control group on days 2 and 4 (P > 0.05); There was no difference among the 10g/L, 50g/L and 200g/L IGF-groups (P > 0.05). On the second day, the proliferative rate of tenocytes in the positive control group was significantly higher than that in 1g/L and 5g/L IGF-groups but lower than in 10g/L, 50g/L and 200g/L IGF-groups (P < or < 0.006). On the fourth day, the proliferative rate of tenocytes in the positive control group was significantly higher than that in each IGF-group and negative control group (P < or < 0.006). On days 2 and 4, the proliferative rate of tenocytes in 1g/L and 5g/L IGF-groups was significantly lower than that in 10g/L, 50g/L and 200g/L IGF-groups (P < 0.000); the positive control group was remarkably higher than the negative control group (P < 0.000). CONCLUSION: IGF-promotes the proliferation of tenocytes in a dose-dependent manner. 10g/L is found to be the most effective concentration. Meanwhile, the proliferative rate of tenocytes is accelerated when the cells are cultured in higher concentration of FCS. 1 Department of Orthopedics, Tianjin Haihe Hospital, Tianjin , China; 2 Tianjin People's Hospital, Tianjin , China; 3 Tianjin Hospital, Tianjin , China Qiu Nan-hai, Master, Attending physician, Department of Orthopedics, Tianjin Haihe Hospital, Tianjin , China qiunanhai@126.com Received: Accepted: Qiu NH, Xia YP, Li MX, Li RH, Kan SL.Dose-effect relationship of insulin-like growth factor and the proliferation of tenocytes in tissue engineering.zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(2): (China) [ / d g/l g/l 5 g/l (P > 0.05)10 g/l 50 g/l 200 g/l (P > 0.05) 2 1 g/l 5 g/l 10 g/l 50 g/l 200 g/l (P < < 0.006) 4 (P < < 0.006) 2 4 1g/L 5g/L 10g/L 50g/L 200g/L (P < 0.000)(P < 0.000) 10 g/l /.[J] (2): [ qiunanhai@126.com :R329.4 :A : (2008) ( /W A) ISSN CN /R CODEN: ZLKHAH 221

2 >><< 10 g/l 10 g/l 2 0 DNAIB DNA B BDNAI / d 10 TBDSCIENCE SIGMA SIGMASIGMA Hanks 10 min Hanks10 ml 10 ml 160 r/min40 min 1mL0.1 5 ml, r/min5 min 5 ml L 20 L10 L 10 / =4/ ml CO 2 80% 1 g/l 10 ml 50 ml 21 g/l 0.15 ml L L g/l P.O.Box 1200,Shenyang kf @sina.com

3 mlhanks 13~ min x _ s SPSS g/l g/l (P < 0.000< 0.006)4 (P < 0.000< 0.006) g/l g/l (P < 0.000) Table MTT test of tenocytes cultured with insulin-like growth factor at different concentrations on days 2 and 4 x _ s Group 2 nd day 4 th day Insulin-like growth factor 1 g/l 5 g/l 10 g/l 50 g/l 200 g/l Positive Negative ab ac ac a ab ac ac a a P0.000, vs. 1 g/l group, 5 g/l group and negative group; b P 0.006, c P=0.000, vs. positive group a: First day b: Second day a: 1 g/l b: 5 g/l c: Thind day d: Seventh day Figure 1 1 Tenocyte apperance and reproduction (200) (200) c: 10 g/l d: 50 g/l g/l P > g/l200 g/l P > Figure 2 2 e: 200 g/l g: Negative control f: Positive control MTT test of cells after insulin-like growth factor is added in the medium for 2 days(100) 2 ISSN CN /R CODEN: ZLKHAH 223

4 a: 1 g/l b: 5 g/l c: 10 g/l d: 50 g/l 1976Grcenlee [1] [2] e: 200 g/l f: Positive control a: 1 g/l b: 5 g/l Figure 3 3 g: Negative control MTT test of cells after insulin-like growth factor is added in the medium for 4 days(100) 4 (100) c: 10 g/l d: 50 g/l 10 g/l 6 1 g/l5g/l10 g/l 50 g/l200 g/l4 e: 200 g/l f: Positive control Figure 4 4 g: Negative control Swiss-gimmsa taint test of cells after insulin-like growth factor of different concentrations is added (100) (100) P.O.Box 1200,Shenyang kf @sina.com

5 [3] [4] [5],: Redaelli [6] DNA [7] mrna [8] mrna [9] [10] [11] [12] [13] [14] 10 g/l 10 g/l [15] g/l [16] [17] [18] [19] nm ISSN CN /R CODEN: ZLKHAH 225

6 H [20] Zhang AYPham HHo Fet alinhibition oftgf-beta-induced collagen production in rabbit flexor tendonsj Hand Surg (2) Zhai HL,Xu F,Cao DJ,et al.shanghai Dier Yike Daxue Xuebao 2004;24(4): [J].,2004,24(4): Zhang QF,Yang ZM,Peng WZ. Zhongguo Xiufu Zhongjian Waike Zazhi 1997;11(1): [J] (1): Xiang Z Yang ZM,Xia QJ,et al. Zhongguo Xiufu Zhongjian Waike Zazhi 1997;11(6):369.mRNA [J] (6):369 5 Liu W, Chen B, Deng D,et al. Repair of tendon defect with dermal fibroblast engineered tendon in a porcine model.tissue Eng 2006;12(4): Redaelli AVesentini SSoncini Met a1possible role of decorin glycosaminoglycans in fibril to fibril force transfer in relative mature tendons -a computational study from molecular to microstructoral levej Biomech 2003;36(10) Yang ZM,Xiang Z,Zou LQ.Zhongguo Xiufu Zhongjian Waike Zazhi 1997;11(5): [J] (5): Zhai HL.Guowai Yixue:Shengwu Yixue Gongcheng Fenche 2004;27(2): [J].,200427(2): Xu F,Liu W,Cao DJ,et al.shanghai Dier Yike Daxue Xuebao 2004;24(4): [J] (4): Xu Y,Tang JB.Zhonghuang Chuangshang Zazhi 2003;19(6): BIB [J]. 2003,19(6): Zou YT,Xiang Z,Yang FC,et al.zhongguo Xiufu Zhongjian Waike Zazhi 2003;17(2): [J] (2): Zhang ZF,Shang QX,Cao YL.Zhongguo Linchuang Kangfu 2002;6(22): [J] (22): Wang W,Wang ZC,Bao YY,et al.diyi Junyi Daxue Xuebao 2001;21(7): [J] (7): Beanes SR, Dang C, Soo C,et a1down-regulation of decorin, a transforming growth factor-beta modulator, is associated with scarless fetal wound healingj Pediatr Surg 2001;36(11): Yang ZM,Xiang Z,Zou LQ,et al. Zhongguo Xiufu Zhongjian Waike Zazhi 1998;12(3): [J] (3): Tsubone T, Moran SL, Amadio PC, et a1expression of growth factors in canine flexor tendon after laceration in vivo.ann Plast Surg 2004;53(4): Harrison RK,Mudem VGrobbelaar AOet a1synovial sheath cell migratory response to flexor tendon injuryan experimental study in ratsj Hand Surg 2003;28(6) Klein MBPham HYalam anchi Net a1flexor tendon wound healin in vitrothe effect of lactate on tendon cell proliferation and collageproductionj Hand Surg Am 2001;26(5) Samiric T, Ilic MZ, Handley CJCharacterisation of proteoglycans and their catabolic products in tendon and explant cultures of tendon.matrix Biol 2004;23(2): Skutek MVan Griensven MZeichen Jet a1cyclic mechanical stretching mod ulates secretion pattern of growt h factors in human tendon fibmblastseurj Appl Physiol 2001;86(1) P.O.Box 1200,Shenyang kf @sina.com

instillation therapy combined with Western medicine can reduce the levels of ALT, TBA, ALP, TBIL, DBIL and GGT, and improve the anti-cmv-igm negative

instillation therapy combined with Western medicine can reduce the levels of ALT, TBA, ALP, TBIL, DBIL and GGT, and improve the anti-cmv-igm negative 在线投稿 : http://www.baishideng.com/wcjd/ch/index.aspx 帮助平台 : http://www.wjgnet.com/esps/helpdesk.aspx DOI: 10.11569/wcjd.v23.i28.4568 世界华人消化杂志 2015 年 10 月 8 日 ; 23(28): 4568-4573 ISSN 1009-3079