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目的:探讨层析法新工艺和原有的低温乙醇工艺制备纤维蛋白原在大鼠跟腱断裂模型中促恢复的效果差异。方法:构建大鼠跟腱部位断裂模型,将其分为空白组(未剪跟腱)、模型组(跟腱断裂未给药)、实验组(自制纤维蛋白原2 mg/mL)与对照组(市售纤维蛋白原2 mg/mL),观察各组大鼠在手术后三周跟腱部位的最大滑动距离、弹性模量和最大抗拉力差异。结果:采用冷沉淀溶解、酸沉除杂、S/D灭活病毒、MacroCap Q柱层析、过滤等流程可从人血浆冷沉淀组分Ⅰ中成功分离纯度为90.9%纤维蛋白原。试验中构建的大鼠跟腱部位断裂模型无感染、且均存活至试验终止。生物力学结果显示,四组大鼠跟腱在最大滑行距离上无明显统计学差异(P>0.05);在跟腱部位弹性模量及最大抗压力比较上,实验组及对照组均优于模型组,但与空白组仍有一定差距,且差异均具有统计学意义(P<0.05)。结论:采用柱层析法分离人纤维蛋白原,不仅能有效提高分离效率,减少蛋白损失,还可增强纤维蛋白原在断裂跟腱中的促恢复效果。
OBJECTIVE: To investigate the differences between the new chromatographic technique and the original low-temperature ethanol preparation for the recovery of fibrinogen in rat Achilles tendon rupture model. Methods: The model of Achilles tendon rupture was established and divided into blank group (without Achilles tendon), model group (not ruptured Achilles tendon), experimental group (homemade fibrinogen 2 mg / mL) and control group (Commercially available fibrinogen 2 mg / mL). The maximal sliding distance, elastic modulus and maximal tensile force of Achilles tendon were observed in each group three weeks after operation. Results: The purity of 90.9% fibrinogen was successfully separated from cryoprecipitate Ⅰ of human plasma by cold sedimentation, acid precipitation, S / D inactivated virus, MacroCap Q column chromatography and filtration. The rat Achilles tendon rupture model was not infected in the experiment and all survived till the end of the experiment. Biomechanical results showed that there was no significant difference in the maximal taxiing distance between the four groups of rats (P> 0.05). Compared with the models of the experimental group and the control group, the elastic modulus and the maximum pressure resistance of the Achilles tendon were superior to those of the model Group, but there was still a gap with the blank group, and the differences were statistically significant (P <0.05). Conclusion: The separation of human fibrinogen by column chromatography can not only effectively improve the separation efficiency, reduce protein loss, but also promote the recovery of fibrinogen in rupture of Achilles tendon.