自体富血小板血浆对颗粒脂肪移植血运重建的影响
发布时间：2018-04-28 05:12
本文选题：富血小板血浆 + 脂肪移植 ； 参考：《大连医科大学》2011年硕士论文
【摘要】：背景:自体脂肪移植由于其组织来源丰富、取材简便、操作简单、充盈外形较好、无排斥反应等优点,被临床上广泛应用,是修饰软组织轮廓缺陷的良好填充材料。富血小板血浆(PRP)是自体血通过梯度离心得到的血小板浓缩物,其内血小板含量丰富,能释放丰富的生长因子。很多研究已经证实PRP中的生长因子对软组织和骨的愈合有促进作用,但是目前还没有PRP对脂肪移植后血运重建影响的报道。 目的:本实验旨在通过建立裸鼠脂肪移植模型,将兔颗粒脂肪中加入自体富血小板血浆混合移植,来探讨富血小板血浆对颗粒脂肪移植后血运重建的影响。 方法:健康新西兰大白兔6只各心脏采血25ml置于预选装有EDTA抗凝剂的试管中,取3ml制备全血血浆,另取2ml用于全血血小板计数,其余20ml制备富血小板血浆。每管10ml置于2个离心管中,2500r/min离心10分钟,吸上清、中间层及红细胞下层1-2mm,移至另一离心管,摇匀。3200 r/min离心8分钟,最下层1ml即为PRP。取PRP0.3ml进行血小板计数,余下移至含有凝血酶及10%氯化钙混合物的管中,4℃冰箱过夜,待血凝块充分收缩后,4500 r/min离心8分钟,取上清,-20℃保存备用。同时无菌条件下取兔颈背部区脂肪垫,浸泡在PBS缓冲液中,清除外包膜、明显的结缔组织和肉眼可见的小血管,用剪刀将脂肪组织剪碎成1mm~3大小,再用生理盐水反复冲洗后置入50ml离心管中静置备用。24只雌雄不限的裸鼠,随机分为4组,每组6只鼠随机排序。每只裸鼠颈背部、背部左右两侧为注射点,三点分别注射①、②或③组脂肪混合物。①实验组:颗粒脂肪0.3ml+明胶海绵125mm~3+PRP0.1ml+生理盐水0.1ml;②实验对照组:颗粒脂肪0.3ml+明胶海绵125mm~3+全血血清0.1ml+生理盐水0.1ml;③空白对照组:颗粒脂肪0.3ml+明胶海绵125mm~3+生理盐水0.2ml。在无菌操作台上,固定裸鼠,消毒后在裸鼠背部标记的三点分别做一长约0.2cm切口,皮下钝性剥离出直径约1.5cm的腔隙,分别填入脂肪组织混合物后缝合切口。术后1、2、4、12周分别处死1、2、3、4组各组6只裸鼠。取出移植物置于体积分数10%多聚甲醛缓冲液中固定,经脱水、透明、石蜡包埋。沿标本纵轴制成4μm厚石蜡切片,每个标本切片5张,一张做HE染色,4张做CD34免疫组化。对全血和PRP进行人工计数,Weidner双盲计数法计算每张组织切片的微血管密度,分析结果。所得数据SPSS16.0统计软件进行统计分析,数据用(x|-)±s表示,并以P㩳0.05为有显著性差异。 结果:24只裸鼠均无感染或死亡。全血血小板计数为(380.3±50.6)×10~9/L, PRP血小板计数为(1744.2±599.6)×10~9/L, PRP血小板计数是全血的4.59倍。各个时相的PRP处理组微血管计数均高于全血血清组及生理盐水组,其中1周和2周的差异有显著性差异(P㩳0.05),4周和12周的差异无显著意义(P㧐0.05)。全血血清组与生理盐水组各个时相差异均无显著意义(P㧐0.05)。 结论:富血小板血浆在颗粒脂肪移植初期能够促进组织血运重建。
[Abstract]:Background: autogenous fat transplantation has been widely used in clinic because of its rich tissue sources, simple material selection, simple operation, good filling appearance and no rejection. It is a good filling material for modifying soft tissue contour defects. Platelet-rich plasma (PRP) is a platelet concentrate obtained from autologous blood by gradient centrifugation. Many studies have confirmed that growth factors in PRP promote soft tissue and bone healing, but there are no reports of the effects of PRP on revascularization after fat transplantation. Objective: to investigate the effect of platelet-rich plasma on revascularization after granulocytic fat transplantation in nude mice by adding autologous platelet-rich plasma into rabbit granular fat. Methods: 25ml was collected from each heart of 6 healthy New Zealand white rabbits and placed in a pre-selected test tube with EDTA anticoagulant. The whole blood plasma was obtained from 3ml, then the whole blood platelet count was obtained from 2ml, and platelet-rich plasma was prepared from other 20ml. Each tube was placed in 2 centrifuge tubes and centrifuged for 10 minutes. The upper layer, middle layer and suberythrocyte layer were removed to another centrifuge tube. The centrifugation lasted for 8 minutes. The lowest layer of 1ml was 1ml. PRP0.3ml was taken for platelet count and the rest was transferred to a refrigerator containing thrombin and 10% calcium chloride mixture for the rest of the night. After the clot was fully constricted, 4500 r/min was centrifuged for 8 minutes, and the supernatant was stored at -20 鈩，
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