北京生物医学工程2024,Vol.43Issue(5) :458-463.DOI:10.3969/j.issn.1002-3208.2024.05.003

硬脑膜修复生物膜的大鼠原位脑植入局部神经毒性评价方法

Local neurotoxicity evaluation method of dura mater repairing biomenbrane in rat brain in situ implantation

邵安良 王小蕾 张勇杰 陈亮 李静莉
北京生物医学工程2024,Vol.43Issue(5) :458-463.DOI:10.3969/j.issn.1002-3208.2024.05.003

硬脑膜修复生物膜的大鼠原位脑植入局部神经毒性评价方法

Local neurotoxicity evaluation method of dura mater repairing biomenbrane in rat brain in situ implantation

邵安良 1王小蕾 1张勇杰 2陈亮 1李静莉1
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作者信息

  • 1. 中国食品药品检定研究院 北京 102629
  • 2. 陕西佰傲再生医学有限公司 西安 710024
  • 折叠

摘要

目的 采用大鼠原位脑植入模型评价硬脑膜修复生物膜的局部神经毒性,旨在建立脑部原位植入局部神经毒性评价方法.方法 选择硬脑膜修复生物膜为研究对象,以上市同类产品为对照品,经大鼠原位脑植入,获取包含颅骨、硬脑膜、植入材料及其周围大脑组织的标本.采用苏木精-伊红(hematoxylin-eosin,HE)染色、Fluoro-Jade C(FJC)特殊染色与胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)免疫荧光考察植入物周围脑组织的局部组织学反应、神经元降解退行性变与星形胶质细胞增生来综合评价植入物的潜在神经毒性风险.结果 植入4周、13周、26周后,与对照品相比,供试品的局部组织学反应均为无刺激反应;FJC染色单位面积阳性细胞数量不存在显著性差异(P>0.05),提示供试品的神经元降解程度与对照组相比不存在显著性差异;GFAP染色单位面积阳性细胞数量不存在显著性差异(P>0.05),提示供试品的星形胶质细胞增生程度与对照组相比不存在显著性差异.结论 可以采用HE染色、FJC特殊染色与GFAP免疫荧光染色联合评价硬脑膜修复生物膜经大鼠原位脑植入后的局部神经毒性.

Abstract

Objective To evaluate the local neurotoxicity of dura mater repairing biomembrane in rat brain in situ implantation model,and to establish a method for local neurotoxicity evaluation in brain in situ implantation.Methods Dura mater repairing biomembrane was selected as test sample,while the same kind of product on the market was used as control.After in situ implantation in rat brain,the specimens including skull,dura mater,implants and surrounding brain tissues were obtained.Hematoxylin-eosin (HE) staining,Fluoro-Jade C (FJC) special staining and glial fibrillary acidic protein (GFAP) immuno-fluorescence staining were used to investigate the local histological response of brain tissue around the implants,neuronal degeneration and astrocyte proliferation for evaluating the potential neurotoxic risk of the implants comprehensively.Results After 4 weeks,13 weeks and 26 weeks in rat brain in situ implantation,the results showed there was no stimulation of the local histological response of the sample compared with control;there was no significant difference in the degree of neuronal degradation between experimental group and control (P>0.05);there was no significant difference in the degree of star Gliosis between experimental group and control (P>0.05) .Conclusions HE staining,FJC special staining and GFAP immune-fluorescence staining could be used to evaluate the local neurotoxicity of the dura mater repairing biomembrane after orthotopic brain implantation in rats.

关键词

硬脑膜修复生物膜/大鼠原位脑植入/局部神经毒性/苏木精-伊红染色/Fluoro-Jade/C特殊染色/胶质纤维酸性蛋白免疫荧光染色

Key words

dura mater repairing biomembrane/rat brain in situ implantation/local neurotoxicity/HE staining/FJC special staining/GFAP immuno-fluorescence

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基金项目

中检院中青年基金(2019C5)

出版年

2024
北京生物医学工程
北京市心肺血管疾病研究所

北京生物医学工程

CSTPCD
影响因子:0.474
ISSN:1002-3208
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