4种冷冻保护剂对人指固有神经的玻璃化保存效果  

Experimental Study on Cryopreservation of Human Proper Digital Nerve by Vitrification with Four Cryoprotective Agents Abstract

作　　者：李智 戴依娜 王成 陈世玖 LI zhi;DAI Yina;WANG Cheng(Department of Plastic and Hand Surgery,The Fifth Affiliated Hospital of Zunyi Medical University,Zhu-hai,Guangdong 519100,China)

机构地区：[1]遵义医科大学第五附属(珠海)医院烧伤整形手外科,519100

出　　处：《医学研究杂志》2022年第7期105-108,共4页Journal of Medical Research

基　　金：广东省珠海市医学科研基金资助项目(20191208A010071)。

摘　　要：目的 评估二甲基亚砜(dimethyl sulfoxide,DMSO)、甘油、丙二醇、乙二醇玻璃化法保存人指固有神经中的保护效果,筛选合适的人指固有神经冷冻保护剂。方法 取截肢手术后患者放弃肢体的指固有神经,分为实验组(30%DMSO组、甘油组、乙二醇组、丙二醇组)和空白对照组进行玻璃化法于液氮(-196℃)中保存3周。复温后进行光学显微镜观察、共聚焦显微镜(laser scanning confocal microscope,LSCM)检测、指固有神经片段的神经生长因子(nerve growth factor,NGF)检测。结果 光镜观察:空白对照组神经鞘膜结构模糊,神经纤维排列疏松、紊乱;实验组神经鞘膜较为完整,神经纤维排列相对整齐、紧凑。LSCM结果:采用钙黄绿素-AM (calcein-AM)和碘化丙啶(propidium iodide,PI)双荧光染色后空白对照组神经纤维绿色荧光强度较弱,红色荧光强度较强。实验组绿色荧光较空白对照组均增强,红色荧光强度均减弱。ELISA检测结果:空白对照组12.95±3.03pg/ml、乙二醇112.64±3.54pg/ml、30%DMSO109.51±9.98pg/ml、甘油61.47±3.34pg/ml、丙二醇62.53±3.64pg/ml。结论4种冷冻保护剂对人指固有神经的玻璃化法保存均有保护作用,其中乙二醇效果最佳,更适合用于人指固有神经玻璃化保存。Objective To evaluate the protective effect of dimethyl sulfoxide(DMSO),glycerol,propylene glycol and ethylene glycol vitrification in the preservation of human finger proper nerve,and to screen a suitable cryoprotective agent for human finger proper nerve.Methods The proper digital nerves of the limbs abandoned after amputation were divided into experimetal group(30% DMSO group,glycerol group,ethylene glycol group,propylene glycol group) and blank control group.They were vitrified and stored in liquid nitrogen(-196℃) for 3 weeks.After rewarming,optical microscope observation,confocal microscope(LSCM) detection and nerve growth factor(NGF) detection of finger intrinsic nerve segments were carried out.Results Light microscope observation:in the blank control group,the structure of nerve sheath was fuzzy,and the arrangement of nerve fibers was loose and disordered;the nerve sheath in the experimental group was relatively complete,and the nerve fibers were arranged relatively orderly and compact.LSCM results:After calcein AM/PI double staining,the green fluorescence intensity of nerve fibers in the blank control group was weak and the red fluorescence intensity was strong;the green fluorescence of the experimental group was stronger than that of the blank control group,and the red fluorescence intensity was weaker.ELISA results:In blank control group,the results were 12.95±3.03 pg/ml,ethylene glycol 112.64±3.54 pg/ml,30% DMSO 109.51±9.98 pg/ml,glycerol 61.47±3.34 pg/ml,propylene glycol 62.53±3.64 pg/ml.Conclusion The four cryoprotectants have protective effects on the vitrification preservation of human finger proper nerve,and ethylene glycol is the best,which is more suitable for the vitrification preservation of human finger proper nerve.

关 键 词：冷冻保护剂 人指固有神经 周围神经损伤 玻璃化保存 

分 类 号：R658[医药卫生—外科学]