本文關(guān)鍵詞： 人臍帶間充質(zhì)干細胞 火器傷 創(chuàng)傷愈合　出處：《福建醫(yī)科大學》2013年碩士論文　論文類型：學位論文

【摘要】：目的： 1.探討人臍帶間充質(zhì)干細胞（human umbilical cord-derived mesenchymalstem cells，hUCMSCs）體外分離、純化和培養(yǎng)方法，并鑒定其生物學特性，為后續(xù)研究奠定基礎(chǔ)。 2.采用不同投射物對豬后肢肌肉軟組織進行致傷，評估、建立一種局部傷情穩(wěn)定性高、可重復性好、實戰(zhàn)模擬性強的火器傷動物模型，為后續(xù)研究奠定基礎(chǔ)。 3.提出一種新的火器傷治療方法：局部清創(chuàng)引流+局部注射hUCMSCs，并在豬后肢火器傷模型中評價其療效。 方法： 1.采用組織塊貼壁法從健康產(chǎn)婦臍帶中分離hUCMSCs，傳代培養(yǎng)后光鏡下進行細胞形態(tài)學觀察，通過流式細胞儀檢測細胞表面抗原，并檢測hUCMSCs成脂、成骨、成軟骨誘導分化的潛能。 2.分別采用“五六”式彈道槍發(fā)射7.62mm制式彈及“五三”式滑膛槍發(fā)射鋼制球形破片射擊8只隨機分配的長白豬雙側(cè)后肢，通過觀察傷情、傷道入出口面積、傷道最大橫截面積、傷道組織病理學分區(qū)，計算傷道組織細菌計數(shù)來評價模型傷情的穩(wěn)定性。 3.取長白豬20只，7.62mm制式彈致傷后隨機分為2組：手術(shù)清創(chuàng)引流+hUCMSCs注射組（A組）、手術(shù)清創(chuàng)引流+無菌生理鹽水注射組（B組）。通過檢測傷道組織細菌量、觀察組織病理學改變以及傷道愈合情況來評價各組的創(chuàng)傷愈合效果。 結(jié)果： 1.組織塊貼壁法分離的hUC-MSCs強表達CD90、CD29、CD73、CD105，弱表達CD106，不表達CD45、CD34及HLA-DR，，能夠分化為脂肪細胞、骨細胞和軟骨細胞。 2.兩種投射物均可造成傷情穩(wěn)定且具有良好可重復性的單純軟組織貫通傷，兩組間傷道入出口面積無顯著差異（p＞0.05），但傷道最大橫截面積有顯著差異（p0.05），兩種方法建立的動物模型均具有典型火器傷病理學特點，細菌計數(shù)檢測顯示傷道組織污染程度均較重，兩組間無顯著差異（p＞0.05）。 3.致傷后經(jīng)不同處理， A組的傷道組織細菌量顯著低于B組（p＜0.05）。兩組間震蕩區(qū)病理學表現(xiàn)不同，Ａ組炎癥反應輕于Ｂ組，A組傷道愈合效果優(yōu)于B組。 結(jié)論： 1.組織塊貼壁法是一種理想的hUC-MSCs體外分離培養(yǎng)方法，根據(jù)這種方法培養(yǎng)出的hUC-MSCs純度高、生物學特征穩(wěn)定，可用于后續(xù)實驗研究。 2.2種投射物致傷效果相似，相比之下，采用“五六”式彈道槍發(fā)射7.62mm制式彈更適合模擬戰(zhàn)時真實傷情，適用于進一步的實驗研究。 3. hUC-MSCs治療肢體火器傷有效。
[Abstract]:Objective:. 1. To investigate the methods of isolation, purification and culture of human umbilical cord-derived mesenchymalstem cells (hUCMSCs) from human umbilical cord mesenchymal stem cells in vitro. 2. Using different projectiles to injure the muscle and soft tissue of pig hind limb, a kind of animal model of firearm injury with high stability, reproducibility and simulated combat was established, which laid a foundation for further study. 3. A new method for the treatment of firearm injury: local debridement and drainage, local injection of hUCMSCsand evaluation of its curative effect in the model of pig hind limb firearm injury. Methods:. 1. HUCMSCs were isolated from umbilical cord of healthy parturient by tissue mass adherent method. Cell morphology was observed under light microscope after passage culture, cell surface antigen was detected by flow cytometry, and the potential of hUCMSCs fat-forming, osteogenesis and chondrogenic differentiation was detected. 2. Using "56-type ballistic gun" to fire 7.62mm standard projectile and "53-type" shotgun firing steel spherical fragment to shoot 8 randomly allocated pig hind limbs. By observing the injury condition, the wound path entrance and exit area and the maximum cross section area of the wound path were observed. To evaluate the stability of the injury model, the bacterial count of wound tissue was calculated in order to evaluate the stability of the model. 3. Twenty Landrace pigs were randomly divided into two groups after injury with 7.62mm standard bomb: surgical debridement and drainage of hUCMSCs injection group (group A) and surgical drainage of sterile saline injection group (group B). Histopathological changes and wound healing were observed to evaluate the wound healing effect in each group. Results:. 1. HUC-MSCs isolated by tissue mass adherent method strongly expressed CD90, CD29, CD73, CD105, weakly expressed CD106, and did not express CD45, CD34 and HLA-DR.It could differentiate into adipocytes, osteocytes and chondrocytes. 2. Both projectiles can cause stable injuries and simple soft tissue penetrating injuries with good repeatability. There was no significant difference in the entrance and exit area between the two groups (p > 0.05), but there was a significant difference in the maximum cross-sectional area of the wound path between the two groups. The animal models established by the two methods had the characteristic of typical firearm wound pathology, and the bacterial count test showed that the wound tissue was polluted seriously. There was no significant difference between the two groups (P > 0.05). 3.The amount of bacteria in wound canal of group A was significantly lower than that of group B (P < 0.05), and the inflammatory reaction of group A was lighter than that of group B (P < 0.05), and the effect of wound healing in group A was better than that in group B. Conclusion:. 1. Tissue mass adherence method is an ideal method for isolation and culture of hUC-MSCs in vitro. The hUC-MSCs obtained by this method has high purity and stable biological characteristics, and can be used for further experimental study. 2.2 the effect of projectile injury is similar. By contrast, the 7.62mm projectile with "five or six" ballistic gun is more suitable for simulating real injury in combat, and suitable for further experimental study. 3. HUC-MSCs is effective in the treatment of firearm injury.
【學位授予單位】：福建醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R82

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