19%浓度DMSO灌注兔离断后肢不同冷冻复温方法骨骼肌微观形态学的研究

发布时间：2018-06-18 21:52

本文选题：深低温保存 + 二甲基亚砜　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:本实验用19%(v/v)的DMSO灌注兔离断后肢,用4种不同冷冻复温方法,对复合组织中的骨骼肌组织损伤程度进行对比研究,找到损伤程度最轻的冷冻复温方法。方法:25只健康的新西兰大白兔随机分为对照组(A组)及实验组(B、C、D、E组,B组为慢冻快复组,C组为慢冻慢复组,D组为快冻慢复组,E组为快冻快复组),实验所用后肢均在髋关节处离断。对照组(A组)后肢在离断后直接用于制作标本。实验组(B、C、D、E组)先用19%浓度DMSO(二甲基亚砜)灌注,肝素盐水洗脱。然后B组慢速冷冻—快速复温、C组慢速冷冻—慢速复温、D组快速冷冻--慢速复温、E组快速冷冻—快速复温,然后采取与对照组相同方法制作标本,病理切片用HE染色后观察骨骼肌损伤程度。根据病理染色的结果,对骨骼肌组织的损伤程度进行统计学分析。结果:经苏木精-伊红(HE)染色后的骨骼肌病理切片为:对照组(A组):骨骼肌细胞核及细胞形态正常,骨骼肌组织未见肌纤维破坏及空泡形成;实验B组:肌细胞肿胀,可见肌纤维断裂,组织形态尚正常;实验C组:肌细胞水肿较轻,有少量空泡,部分肌纤维断裂;实验D组:肌细胞水肿严重,少量空泡,部分肌纤维断裂;实验E组:肌细胞肿胀明显,大量空泡化,肌纤维断裂明显,肌纤维部分溶解。结论:1.在19%(v/v)二甲基亚砜灌注条件下,实验组(B、C、D、E组)和对照组(A组)病理切片结果之间存在显著性差异,这表明深低温冷冻复温对细胞有损伤;2.各个实验组之间的结果差异有统计学意义,说明骨骼肌损伤程度因冷冻复温方式不同而不同;3.实验组骨骼肌损伤程度由轻到重为:慢速冷冻-快速复温组慢速冷冻-慢速复温组快速冷冻-慢速复温组快速冷冻-快速复温组,可以认为在四种冷冻复温方法中,慢速冷冻-快速复温是最佳的冷冻复温方法。
[Abstract]:Objective: in this experiment, 19% (v/v) DMSO was used to perfusion the hind limbs of rabbits, and 4 different cryopreservation methods were used to compare the damage degree of skeletal muscle tissue in the composite tissue, and to find the lightest degree of cryopreservation. Methods: 25 healthy New Zealand white rabbits were randomly divided into control group (group A) and experimental group (group B, C, D, E group, B group) Group C was slow freezing fast recovery group, group C was slow freezing slow recovery group, group D was fast freezing slow recovery group, group E was fast freeze-drying group, and the hind limbs were disconnected at hip joint. The hind limbs of control group (group A) were directly used to make specimens after separation. The experimental group (B, C, D, E group) was perfused with 19% concentration DMSO (two methyl sulfoxide) and heparin brine eluted. Then B group slow freezing quick freezing fast At the rapid recovery temperature, the C group slow freezing - slow rewarming, the D group fast freezing - slow rewarming, the E group fast freezing - fast rewarming, and then taking the same method as the control group to make the specimens. The pathological section was stained with HE to observe the degree of skeletal muscle injury. The damage degree of the bone muscle tissue was statistically analyzed according to the pathological staining. Results: The pathological sections of skeletal muscle after hematoxylin eosin (HE) staining were as follows: control group (group A): the nucleus and cell morphology of skeletal muscle were normal, muscle fibers were not damaged and vacuoles formed in skeletal muscle tissue; experimental B group: muscle cell swelling, muscle fiber rupture, and normal tissue morphology; experimental C group: muscle cell edema was lighter, a small amount of vacuoles, partial muscles were found. Fiber rupture; experimental group D: muscle cell edema, a small number of vacuoles, and partial muscle fiber rupture; experimental group E: muscle cells swelling obviously, a large number of vacuolation, muscle fiber fracture obvious, muscle fiber partial dissolution. Conclusion: 1. in the 19% (v/v) two methyl sulfoxide perfusion conditions, the test group (B, C, D, E group) and the control group (A group) pathological section between the results exist between the pathological section between the results exist between the pathological sections (A group) between pathological sections exist between the results of the presence of the obvious between the group (A group) pathological section between the results exist between the obvious This indicates that cryopreservation can be damaged by cryopreservation; 2. the difference between the experimental groups is statistically significant, indicating that the degree of skeletal muscle injury is different because of the different freezing methods. The degree of skeletal muscle injury in the 3. experimental group is from light to heavy: slow freezing fast rewarming group fast freezing and slow rewarming group - slow rewarming group rapid freezing rapid rewarming group, it can be considered that in the four refrigeration rewarming methods, slow freezing rapid rewarming is the best method of freezing rewarming.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R658

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