玻璃化法与化学萃取法制备同种异体肌腱的生物学性能比较

The comparison study on the biological performances of allogeneic tendon prepared by vitrification and chemical extraction method

目的比较玻璃化法与化学萃取法制备同种异体肌腱的生物学性能。方法选取6个月月龄白色健康雄性来亨鸡48只,随机分为3组,每组16只。A组(玻璃化组)、B组(化学萃取组)和C组(空白对照组),分别进行组织形态学观察、生物力学测试和免疫原性检测。结果 (1)两种方法制备的肌腱与空白对照组肌腱在长度和横截面积上无明显变化,其外在结构保持一致。A、C组肌腱在细胞数目、大小及形态方面相似,B组肌腱几乎无腱细胞残存;(2)3组间的拉伸断裂强度(Pmax)、拉伸断裂功耗(Wmax)和拉伸断裂延伸率(&max)的结果差异无统计学意义(P>0.05);(3)3组间1、2周末CD+4、CD+8T淋巴细胞含量及CD+4/CD+8的比值差异有统计学意义(P<0.05);3、6周末A、B组间CD+4、CD+8T淋巴细胞含量及CD+4/CD+8的比值差异无统计学意义(P>0.05),AC、BC组间差异有统计学意义(P<0.05)。结论玻璃化法较化学萃取法制备的肌腱保留了部分原有肌腱细胞;玻璃化法和化学萃取法制备的肌腱明显降低了肌腱的免疫原性,均保留了良好的生物力学性能。

Objective To compare the biological performances of allogeneic tendon prepared by vitrification and chemical extraction method. Methods Forty-eight white healthy male Leghorns （6-month age） were randomly divided into three groups,group A （vitrification group）, group B （chemical extraction group） and group C （blank control group）, with 16 Leghorns in each group. The histo-morphological observation, biomechanics test and immunogenicity detection for the allogeneic tendon prepared by the two methods were carried out, respectively. Results There were no significant changes in length and cross sectional area of the allogeneic tendon prepared by both methods, as compared with those in blank control group, with consistent external structures. There were no significant differences in the cell number, size, shape of tendons between group A and group C, however, there was nearly no tendon cell remnant in group B. There were no significant differences in the tensile fracture strength （Pmax）, tensile fracture power consumption （Wmax） and tensile elongation at rupture rate （＆max） among the three groups （ P 〉 0.05 ）. However there were significant differences in the contents of CD4 , CDs＋ T lymphocytes and the ratio of CD4/CDs＊ at the end of 1,2 weeks among three groups （ P 〈0.05） ,and there were no significant differences in the contents of CD4, CDs＋ T lymphocytes and the ratio of CD4/CDs＋ at the end of 3, 6 weeks between group A and group B （ P 〉 0.05 ）, but there were significant differences between group A and group C and between group B and group C （ P 〈 0.05）. Conclusion Vitrification method is superior to chemical extraction method in the preparation of tendon, which retains original tendon cells, and both methods obviously reduce the immunogenicity of tendon and retain favourable biomechanics performance.