MSCs附和PLGA電紡支架修復兔跟腱損傷的實驗研究
發(fā)布時間:2018-04-19 17:30
本文選題:骨髓間充質干細胞 + 靜電紡絲; 參考:《中國人民解放軍軍醫(yī)進修學院》2012年博士論文
【摘要】:目的:研究兔骨髓間充質干細胞附和PLGA電紡支架修復跟腱損傷的作用。 方法:利用靜電紡絲技術制備PLGA電紡絲膜狀支架,對PLGA支架結構進行觀察和測量,并使用高溫高壓、低溫等離子、鉆6。照射、紫外線照射、酒精浸泡等五種不同方式消毒,進行細菌培養(yǎng)檢測消毒后效果并觀察支架結構的破壞情況;采用骨髓穿刺法獲取兔骨髓,并用密度梯度離心法分離并體外培養(yǎng)骨髓間充質干細胞(MSCs);將第3代骨髓間充質干細胞結合PLGA支架復合培養(yǎng);將45只成年新西蘭大白兔,隨機分為3組,每組15只,將左后肢跟腱中央束切斷,改良Kessler法縫合,MSCs組使用MSCs附和PLGA電紡絲膜進行包裹,PLGA支架組使用單純PLGA電紡絲膜進行包裹,對照組只做單純縫合。分別在2、4、6周時間點各組處死5只兔取材,組織學和生物力學分析評估跟腱的愈合情況。 結果:PLGA電紡絲膜狀支架纖維直徑均勻,結構規(guī)整,孔隙率82.6±3.45%;五種消毒方式均可達到滅菌目的;其中鉆6。照射和紫外線照射對支架結構的破壞最小最適合PLGA電紡絲膜狀支架的消毒;使用梯度離心法分離獲得的MSCs純度高,生物學特性明顯,體外增殖能力較強,可以與PLGA電紡絲膜狀支架較好結合;MSCs組和PLGA電紡支架組組織學顯示肌腱愈合好,與周圍組織粘連較輕,生物力學優(yōu)于對照組。 結論:靜電紡絲技術是獲得納米級纖維的一種簡便方法,使用梯度離心法分離獲得的MSCs純度較高,生物學特性明顯,體外增殖能力較強,可以與PLGA電紡絲膜狀支架結合;MSCs附和PLGA電紡支架能夠促進兔跟腱損傷的愈合,減少外源性疤痕組織長入,減少肌腱與周圍組織的粘連。
[Abstract]:Aim: to study the effect of rabbit bone marrow mesenchymal stem cells (BMSCs) and PLGA electrospun scaffold on the repair of Achilles tendon injury.Methods: the PLGA electrospun membrane scaffolds were prepared by electrospinning technique. The structure of PLGA scaffolds was observed and measured. High temperature and high pressure, low temperature plasma, drilling 6.Five different disinfection methods, such as irradiation, ultraviolet radiation and alcohol immersion, were carried out to detect the disinfection effect and observe the destruction of the scaffold structure, and the rabbit bone marrow was obtained by bone marrow puncture.Bone marrow mesenchymal stem cells (MSCs) were isolated by density gradient centrifugation and cultured in vitro. Bone marrow mesenchymal stem cells (BMSCs) of the third generation were co-cultured with PLGA scaffolds. 45 adult New Zealand white rabbits were randomly divided into 3 groups, 15 in each group.The left hind limb was cut off from the central tendon of the Achilles tendon. The modified Kessler method was used to sew the MSCs and PLGA electrospinning membrane. The PLGA electrospun membrane was used to wrap the scaffold group, while the control group was sutured only.Five rabbits were sacrificed at the time of 2 weeks and 4 weeks, respectively. The healing of Achilles tendon was evaluated by histological and biomechanical analysis.Results the diameter of the fiber was uniform, the structure was regular, the porosity was 82.6 鹵3.45, and all the five disinfection methods could be used to sterilize the fibers.The minimal damage to the scaffold structure caused by irradiation and ultraviolet irradiation is the most suitable for disinfection of PLGA electrospinning membrane scaffold. The MSCs isolated by gradient centrifugation has high purity, obvious biological characteristics and strong proliferative ability in vitro.It could be combined with PLGA electrospinning membrane scaffold and PLGA electrospinning scaffold to show that tendon healed well, adhesion to surrounding tissue was lighter, biomechanics was superior to control group.Conclusion: electrostatic spinning is a simple and convenient method to obtain nanofibers. MSCs obtained by gradient centrifugation has high purity, obvious biological characteristics and strong proliferative ability in vitro.PLGA and PLGA can promote the healing of Achilles tendon injury, reduce the growth of exogenous scar tissue and reduce the adhesion between tendon and surrounding tissues.
【學位授予單位】:中國人民解放軍軍醫(yī)進修學院
【學位級別】:博士
【學位授予年份】:2012
【分類號】:R318.08
【參考文獻】
相關期刊論文 前10條
1 鄒云雯,夏精武,季愛玉,葉發(fā)剛,齊宗華,王鐵丹,肖應慶,邢立峰;人發(fā)角蛋白人工腱的臨床應用[J];青島醫(yī)學院學報;1997年03期
2 龍劍虹,祁敏,黃曉元,雷少榕,任利成;膠原-聚羥基乙酸與骨髓間質干細胞的細胞相容性研究[J];中國醫(yī)師雜志;2005年02期
3 周強,李起鴻,楊柳,段小軍;骨髓間充質干細胞在軟骨細胞培養(yǎng)條件下生物學特點的觀察[J];第三軍醫(yī)大學學報;2002年05期
4 賈駿;段Z腪,
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