摘要
目的:制备去细胞周围神经移植物并评测其生物相容性。方法:实验于2005-05/12在解放军第四军医大学口腔医学院组织病理教研室和组织工程实验中心完成。①将猪肋间神经经低渗、-80℃反复冻融和NaOH裂解等方法制备去细胞神经异体移植物;光、电镜观察其结构特征。②材料细胞毒性试验按照GB/T16886.5-2003《医疗器械生物学评价》体外细胞毒性试验的试验方法进行,毒性程度的评估标准参见美国药典。③大鼠皮下植入试验,取成年SD大鼠9只,于脊柱中线两侧约2cm处平行各取2点作为皮下植入点,植入1cm去细胞异体神经移植物。于手术后第1,2,4周时各取1只大鼠麻醉状态下处死,作组织切片检查。结果:去细胞神经异体移植物的制备:①应用正常猪的周围神经,清除许旺细胞、神经外膜和束膜细胞以及神经纤维的髓鞘和轴突,保存由许旺细胞基底膜管以及神经外膜和束膜的细胞外基质构成的三维支架结构。②生物相容性测试结果:显示去细胞神经异体移植物细胞毒性试验为1级。③SD大鼠皮下植入试验结果:去细胞神经异体移植物无明显的免疫排斥反应。结论:该方法所制备的去细胞神经移植物神经膜管结构保存完好,有良好的组织相容性,可能成为自体神经移植的替代材料。
AIM： To prepare acellular peripheral nerve graft and evaluate its biocompatibility.
METHODS： The experiment was carried out in the Department of Oral Histology and Pathology, and Tissue Engineering Center, Stomatological College of the Fourth Military Medical University of Chinese PLA from May to December 2005.①The acellular nerve allografts were prepared with fresh pig intercostal nerve by hypotension, freezing-thawing at -80℃ repeatedly and NaOH splitting; And the characteristics of structure were observed under light and electron microscope. ② The cytotoxicity of acellular nerve was tested according to GB/T16886.5-2003 Biological evaluation of medical devices： Tests for cytotoxicity in vitro, and the levels of cytotoxicity were evaluated with U.S. Pharmacopeia. ③.Subdermal implantation in rats： Nine adult SD rats were assigned to select two implanted points parallel in 2 cm distance from spinal middle line for acellular nerve allograft. Three rats were executed respectively on postoperative 1, 2 and 4 weeks to conduct tissue section examination.
RESULTS： ①Preparation of acellular nerve allografts： Schwann cells, the cells of epineurium or perineurium, myelin sheath and axon of nerve fibers were eliminated from normal pig peripheral nerves. However, the three dimensional supports of basement membrane tube of Schwann cells and extracellular matrix of epineurium and perineurium were reserved. ②The cytotoxicity of acellular nerve allograft was proved to be first grade by biocompatibility test.③Tbere was no obvious immunologic rejection reaction of the acellular nerve allografts in subdermal implantation of SD rats.
CONCLUSION： The acellular nerve grafts possess well-conserved structure of neurilemma tube and excellent histocompatibility, and is the possible replaced material for autologous nerve graft.
出处
《中国临床康复》
CSCD
北大核心
2006年第21期82-84,共3页
Chinese Journal of Clinical Rehabilitation
