本文選題：富血小板血漿 + 脂肪移植��； 參考：《大連醫(yī)科大學》2011年碩士論文

【摘要】：背景:自體脂肪移植由于其組織來源豐富、取材簡便、操作簡單、充盈外形較好、無排斥反應等優(yōu)點,被臨床上廣泛應用,是修飾軟組織輪廓缺陷的良好填充材料。富血小板血漿(PRP)是自體血通過梯度離心得到的血小板濃縮物,其內血小板含量豐富,能釋放豐富的生長因子。很多研究已經證實PRP中的生長因子對軟組織和骨的愈合有促進作用,但是目前還沒有PRP對脂肪移植后血運重建影響的報道。 目的:本實驗旨在通過建立裸鼠脂肪移植模型,將兔顆粒脂肪中加入自體富血小板血漿混合移植,來探討富血小板血漿對顆粒脂肪移植后血運重建的影響。 方法:健康新西蘭大白兔6只各心臟采血25ml置于預選裝有EDTA抗凝劑的試管中,取3ml制備全血血漿,另取2ml用于全血血小板計數,其余20ml制備富血小板血漿。每管10ml置于2個離心管中,2500r/min離心10分鐘,吸上清、中間層及紅細胞下層1-2mm,移至另一離心管,搖勻。3200 r/min離心8分鐘,最下層1ml即為PRP。取PRP0.3ml進行血小板計數,余下移至含有凝血酶及10%氯化鈣混合物的管中,4℃冰箱過夜,待血凝塊充分收縮后,4500 r/min離心8分鐘,取上清,-20℃保存?zhèn)溆�。同時無菌條件下取兔頸背部區(qū)脂肪墊,浸泡在PBS緩沖液中,清除外包膜、明顯的結締組織和肉眼可見的小血管,用剪刀將脂肪組織剪碎成1mm~3大小,再用生理鹽水反復沖洗后置入50ml離心管中靜置備用。24只雌雄不限的裸鼠,隨機分為4組,每組6只鼠隨機排序。每只裸鼠頸背部、背部左右兩側為注射點,三點分別注射①、②或③組脂肪混合物。①實驗組:顆粒脂肪0.3ml+明膠海綿125mm~3+PRP0.1ml+生理鹽水0.1ml;②實驗對照組:顆粒脂肪0.3ml+明膠海綿125mm~3+全血血清0.1ml+生理鹽水0.1ml;③空白對照組:顆粒脂肪0.3ml+明膠海綿125mm~3+生理鹽水0.2ml。在無菌操作臺上,固定裸鼠,消毒后在裸鼠背部標記的三點分別做一長約0.2cm切口,皮下鈍性剝離出直徑約1.5cm的腔隙,分別填入脂肪組織混合物后縫合切口。術后1、2、4、12周分別處死1、2、3、4組各組6只裸鼠。取出移植物置于體積分數10%多聚甲醛緩沖液中固定,經脫水、透明、石蠟包埋。沿標本縱軸制成4μm厚石蠟切片,每個標本切片5張,一張做HE染色,4張做CD34免疫組化。對全血和PRP進行人工計數,Weidner雙盲計數法計算每張組織切片的微血管密度,分析結果。所得數據SPSS16.0統(tǒng)計軟件進行統(tǒng)計分析,數據用(x|-)±s表示,并以P㩳0.05為有顯著性差異。 結果:24只裸鼠均無感染或死亡。全血血小板計數為(380.3±50.6)×10~9/L, PRP血小板計數為(1744.2±599.6)×10~9/L, PRP血小板計數是全血的4.59倍。各個時相的PRP處理組微血管計數均高于全血血清組及生理鹽水組,其中1周和2周的差異有顯著性差異(P㩳0.05),4周和12周的差異無顯著意義(P㧐0.05)。全血血清組與生理鹽水組各個時相差異均無顯著意義(P㧐0.05)。 結論:富血小板血漿在顆粒脂肪移植初期能夠促進組織血運重建。
[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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