辐照氟银异体肌腱移植的实验研究
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摘要
目的 观察辐照氟银异体肌腱移植的效果，同时建立一种新的异体肌腱处理技术。方法 32只家兔随机分为4组：分别将制备好的辐照氟银异体肌腱（A组），单纯辐照异体肌腱（B组），深低温冷冻异体肌腱（C组）与自体肌腱（D组），行肌腱移植。术前进行各组微生物学检测；术前、术后对比观察生物力学及形态学组织学的变化；术后行各组免疫源性、羟脯氨酸含量测定。结果 ⑴与A组相比，单纯辐照组及深低温冷冻组抑菌作用极为微弱（P<0.01）。A组含银量高，而其余各组未检出。⑵术前辐照组最大载荷和抗张强度明显大于其它各组（P<0.01），但术后12周差别无统计学意义（P>.05）。⑶组织学变化：术后4周各处理组细胞增多，均多于自体组，A、B组细胞呈椭圆形，C组为线形或梭形；术后12周各处理组细胞减少，仍多于自体组，细胞以线形或梭形为主。⑷辐照组淋巴细胞死亡率与自体组无差异（P>0.05），均低于冷冻组（P<0.05）；术后12周辐照组羟脯氨酸含量与自体组无差异（P>.05），但高于冷冻组（P<0.05）。结论 辐照氟银异体肌腱具有明显的低抗原性、无菌且有强抑菌作用，可作为一种良好的自体肌腱替代材料。
Objective To observe the effect of homograft of γ-Irradiated tendon incorporated with norfloxacin and silver immersion and develop a new homograft tendon processing technique. Methods 32 rabbits were randomly divided into the following four groups: irradiated allogeneic tendon incorporated with norfloxacin and silver group(Group A), irradiated allogeneic tendon group(Group B),deep-frozen allogeneic tendon group(Group C)and autogeneic tendon group(Group D).They were then implanted into defects of Achilles tendons respectively. Ag+ level and bacteriostatic effect were detected in vitro. Biomechanic or histologic properties were observed preoperatively and postoperatilely. Immunologic property and hydroxyproline content were determined postoperatively. Results (1)No bacterial growth was investigated in Group A and B.By contrast,tendons of A group can effectively inhibit growth of bacteria(P<0.01).(2) Before implantation, the ultimate tensile stress of Group A and B was markedly greater than that of Group C and D（P<0.01）,but there was no significant difference 12 weeks after operation.(3)Histological findings:At four weeks after transplantation,there were more cells in all grafts than in normal control tendons.The cells of the irradiated grafts were mainly oblong while others were predominatly spindle-shaped or linear. At 12 weeks,cellularity of all grafts had decreased but remained greater than in normal control tendons.The cells were mainly spindle-shaped or linear .(4) The contents of lymphocytoxicity showed lower in Group A,B and D than in Group C（P<0.05）.The Hyp level was no significant between Group A ,Band D（P>0.05）, but it was lower in Group C（P<0.05）.Conclusion γ-Irradiated tendon incorporated with norfloxacin and silver immersion is a good substitute for autogenous grafts,which can remarkably decreased the antigenecity with sterility and strong bacteriostatic effect.
Objective To observe the effect of homograft of γ-Irradiated tendon incorporated with norfloxacin and silver immersion and develop a new homograft tendon processing technique. Methods 32 rabbits were randomly divided into the following four groups: irradiated allogeneic tendon incorporated with norfloxacin and silver group(Group A), irradiated allogeneic tendon group(Group B),deep-frozen allogeneic tendon group(Group C)and autogeneic tendon group(Group D).They were then implanted into defects of Achilles tendons respectively. Ag+ level and bacteriostatic effect were detected in vitro. Biomechanic or histologic properties were observed preoperatively and postoperatilely. Immunologic property and hydroxyproline content were determined postoperatively. Results (1)No bacterial growth was investigated in Group A and B.By contrast,tendons of A group can effectively inhibit growth of bacteria(P<0.01).(2) Before implantation, the ultimate tensile stress of Group A and B was markedly greater than that of Group C and D（P<0.01）,but there was no significant difference 12 weeks after operation.(3)Histological findings:At four weeks after transplantation,there were more cells in all grafts than in normal control tendons.The cells of the irradiated grafts were mainly oblong while others were predominatly spindle-shaped or linear. At 12 weeks,cellularity of all grafts had decreased but remained greater than in normal control tendons.The cells were mainly spindle-shaped or linear .(4) The contents of lymphocytoxicity showed lower in Group A,B and D than in Group C（P<0.05）.The Hyp level was no significant between Group A ,Band D（P>0.05）, but it was lower in Group C（P<0.05）.Conclusion γ-Irradiated tendon incorporated with norfloxacin and silver immersion is a good substitute for autogenous grafts,which can remarkably decreased the antigenecity with sterility and strong bacteriostatic effect.
引文
1 Seiler JG,Gelberman RH,Williams CS,etal.Autogenous flexor-tendon graft.J Bone Joint Surg(Am),1993,75:1004-1010
2 Kato YP,Dunn MG,Zawadsky JP,etal.Regeneration of Achilles tendon with a collagen tendon prosthesis.J Bone Joint Surg[Am],1991,73(4):561-573
3 Langer R,Vacanti JP.Science,1993,260:920-926
4 闫玉华.组织工程基质研究进展.国外医学:生物医学分册,,4:149-153
5 田立杰，王彦生，战杰,等. 同种异体肌腱移植的临床应用. 中华手外科杂志， 1999，15（2）：93-94
6 唐林俊，雷军，方光荣,等. 超深低温处理的异体肌腱移植. 中华手外科杂志， 2001，17，增刊：1-3
7陈星隆,高伟阳,洪建军,等.普通低温冷冻保存异体肌腱的应用研究. 中华手外科杂志,2001，17（4）：223-224
8 Shino Konsei,Kimura Tomoatsu,Hirose Hitoshi,etal.Reconstuction of the anterior cruciate ligament by allogeneic tendon graft. J Bone Joint Surg[Am].1986,68(5):739-746
9张友乐，杨克非，朱伟,等.异体肌腱移植的实验研究与临床应用.中华外科杂志，1995，33（9）：539-541
10 沈惠良,李旗,张杰,等.深低温保存的异体骨-髌腱-骨重建膝关节前交叉韧带. 中华创伤杂志,2001，17（11）：681-683
11唐林俊，程国良，方光荣，等. 反复冻融及超深低温处理的异体肌腱移植实验研究. 中国修复重建外科杂志，1998，12（4）：241-244
12唐林俊，程国良，方光荣等. 培养处理与冷冻保存同种异体肌腱的实验研究. 解放军医学杂志,1999，24（3）：196-199
13 鲁晓波,杨志明.冷冻小鼠 同种异体肌腱移植后IL-2生物活性水平的检测. 中华手外科杂志,1999，15（1）：56-58
14 Shoskes DA, Wood KJ. Indirect presentation of MHC antigens in transplantation. Immunol Today. 1994,15(1):32-40
15 Guidos C, Sinha AA, Lee KC. Functional differences and complementation between dendritic cells and macrophages in Tcell activation. Immunology. 1987,61:269-276
16 夏惠生,主编.器官移植学.第一版.上海:上海科技出版社,1995.43
17鲁晓波,杨志明.冷冻同种异体肌腱小鼠移植的细胞毒、IL-2水平组织学观察. 中华手外科杂志,1999，15（4）：275-277
18 Yukiyoshi Toritsuka, Konsei Shino, Shuji Horibe, etal. Effect of freeze-drying or r-irradiation on remodeling of tendon allograft in rat model. J Orthop Res 1997，15:294-300
19Grant RA,Cox RW,Kent CM.The effects of gamma irradiation on the structure and reactivity of native and cross-linked collagen fibres.J Anat,1973,115:29-43
20BaileyAJ,RhodesDN.Irradiation-inducedcrosslinkingof collagen.Radiat Res, 1964,22:606-621
21Bailey AJ.Effects of ionizing radiation on the ultrastructure of collagen fibrils. Radiat Res,1964,23:145-155
22 Salehpour A,Butler DL,Proch FS,etal.Dose-dependent response of gamma irradiation on echanical properties and related biochemical composition of goat bone-patellartendon-bone allografts.J Orthop Res.1995,13(6):898-906
23 Rasmussen TJ, Feder SM, D.L.Butler, etal. The effects of 4 Mrad of irradiation on the initial mechanical properties of bone-patellar tendon-bone grafts. Arthroscopy 1994,10(2):188-197.
24 Gibbons MJ, Butler DL, Grood ES, etal. Effects of gamma irradiation on the initial mechanical and material properties of goat bone-patellar tendon-bone allografts. J Orthop Res , 1991, 9:209-218
25 Jackson DW,Beach Long,Grood ES,etal.The effects of in situ freezing on the anterior cruciate ligament.J Bone Joint Surg[Am].1991,73(2):201-212
26 Jackson DW,Simon TM,Kurzweil PR,etal.Survival of cells after intra-articular transplantation of fresh allografts of the patellar and anterior cruciate ligaments. J Bone Joint Surg[Am].1992,74(1):112-118
27 Arnoczky SP,Warren RF,Ashlock MA.replacement of the anterior cruciate ligament using a patellar tendon allograft. J Bone Joint Surg[Am].1986,68(3):376-385
28 Horibe S,Shino K,Nagano J,etal.Replacing the medial collateral ligament with an allogeneic tendon graft. J Bone Joint Surg[Am].1990, 72(6):1044-1049
29Akira Maeda,Shuji Horibe,Norinao Matsumoto,etal.Solvent-dried and gamma-irradiated tendon allografts in rats. J Bone Joint Surg[Br].1998, 80-B:731-736
30胡蕴玉。骨库的建立与管理。中华骨科杂志。1995，15（1）：54-58
31李国辉，曹勇，吴燮卿，等.辐照氟银猪皮的实验研究.中华整形烧伤外科杂志，1994，10（3）：206-208
32薛广波主编.实用消毒学.北京:人民军医出版社.1986
2 Kato YP,Dunn MG,Zawadsky JP,etal.Regeneration of Achilles tendon with a collagen tendon prosthesis.J Bone Joint Surg[Am],1991,73(4):561-573
3 Langer R,Vacanti JP.Science,1993,260:920-926
4 闫玉华.组织工程基质研究进展.国外医学:生物医学分册,,4:149-153
5 田立杰，王彦生，战杰,等. 同种异体肌腱移植的临床应用. 中华手外科杂志， 1999，15（2）：93-94
6 唐林俊，雷军，方光荣,等. 超深低温处理的异体肌腱移植. 中华手外科杂志， 2001，17，增刊：1-3
7陈星隆,高伟阳,洪建军,等.普通低温冷冻保存异体肌腱的应用研究. 中华手外科杂志,2001，17（4）：223-224
8 Shino Konsei,Kimura Tomoatsu,Hirose Hitoshi,etal.Reconstuction of the anterior cruciate ligament by allogeneic tendon graft. J Bone Joint Surg[Am].1986,68(5):739-746
9张友乐，杨克非，朱伟,等.异体肌腱移植的实验研究与临床应用.中华外科杂志，1995，33（9）：539-541
10 沈惠良,李旗,张杰,等.深低温保存的异体骨-髌腱-骨重建膝关节前交叉韧带. 中华创伤杂志,2001，17（11）：681-683
11唐林俊，程国良，方光荣，等. 反复冻融及超深低温处理的异体肌腱移植实验研究. 中国修复重建外科杂志，1998，12（4）：241-244
12唐林俊，程国良，方光荣等. 培养处理与冷冻保存同种异体肌腱的实验研究. 解放军医学杂志,1999，24（3）：196-199
13 鲁晓波,杨志明.冷冻小鼠 同种异体肌腱移植后IL-2生物活性水平的检测. 中华手外科杂志,1999，15（1）：56-58
14 Shoskes DA, Wood KJ. Indirect presentation of MHC antigens in transplantation. Immunol Today. 1994,15(1):32-40
15 Guidos C, Sinha AA, Lee KC. Functional differences and complementation between dendritic cells and macrophages in Tcell activation. Immunology. 1987,61:269-276
16 夏惠生,主编.器官移植学.第一版.上海:上海科技出版社,1995.43
17鲁晓波,杨志明.冷冻同种异体肌腱小鼠移植的细胞毒、IL-2水平组织学观察. 中华手外科杂志,1999，15（4）：275-277
18 Yukiyoshi Toritsuka, Konsei Shino, Shuji Horibe, etal. Effect of freeze-drying or r-irradiation on remodeling of tendon allograft in rat model. J Orthop Res 1997，15:294-300
19Grant RA,Cox RW,Kent CM.The effects of gamma irradiation on the structure and reactivity of native and cross-linked collagen fibres.J Anat,1973,115:29-43
20BaileyAJ,RhodesDN.Irradiation-inducedcrosslinkingof collagen.Radiat Res, 1964,22:606-621
21Bailey AJ.Effects of ionizing radiation on the ultrastructure of collagen fibrils. Radiat Res,1964,23:145-155
22 Salehpour A,Butler DL,Proch FS,etal.Dose-dependent response of gamma irradiation on echanical properties and related biochemical composition of goat bone-patellartendon-bone allografts.J Orthop Res.1995,13(6):898-906
23 Rasmussen TJ, Feder SM, D.L.Butler, etal. The effects of 4 Mrad of irradiation on the initial mechanical properties of bone-patellar tendon-bone grafts. Arthroscopy 1994,10(2):188-197.
24 Gibbons MJ, Butler DL, Grood ES, etal. Effects of gamma irradiation on the initial mechanical and material properties of goat bone-patellar tendon-bone allografts. J Orthop Res , 1991, 9:209-218
25 Jackson DW,Beach Long,Grood ES,etal.The effects of in situ freezing on the anterior cruciate ligament.J Bone Joint Surg[Am].1991,73(2):201-212
26 Jackson DW,Simon TM,Kurzweil PR,etal.Survival of cells after intra-articular transplantation of fresh allografts of the patellar and anterior cruciate ligaments. J Bone Joint Surg[Am].1992,74(1):112-118
27 Arnoczky SP,Warren RF,Ashlock MA.replacement of the anterior cruciate ligament using a patellar tendon allograft. J Bone Joint Surg[Am].1986,68(3):376-385
28 Horibe S,Shino K,Nagano J,etal.Replacing the medial collateral ligament with an allogeneic tendon graft. J Bone Joint Surg[Am].1990, 72(6):1044-1049
29Akira Maeda,Shuji Horibe,Norinao Matsumoto,etal.Solvent-dried and gamma-irradiated tendon allografts in rats. J Bone Joint Surg[Br].1998, 80-B:731-736
30胡蕴玉。骨库的建立与管理。中华骨科杂志。1995，15（1）：54-58
31李国辉，曹勇，吴燮卿，等.辐照氟银猪皮的实验研究.中华整形烧伤外科杂志，1994，10（3）：206-208
32薛广波主编.实用消毒学.北京:人民军医出版社.1986