發(fā)布時(shí)間：2018-04-19 17:30

  本文選題：骨髓間充質(zhì)干細(xì)胞 + 靜電紡絲��； 參考：《中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院》2012年博士論文

【摘要】：目的：研究兔骨髓間充質(zhì)干細(xì)胞附和PLGA電紡支架修復(fù)跟腱損傷的作用。 方法：利用靜電紡絲技術(shù)制備PLGA電紡絲膜狀支架,對(duì)PLGA支架結(jié)構(gòu)進(jìn)行觀察和測(cè)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(jìn)行細(xì)菌培養(yǎng)檢測(cè)消毒后效果并觀察支架結(jié)構(gòu)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(zhì)干細(xì)胞(MSCs);將第3代骨髓間充質(zhì)干細(xì)胞結(jié)合PLGA支架復(fù)合培養(yǎng)；將45只成年新西蘭大白兔,隨機(jī)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(jìn)行包裹,PLGA支架組使用單純PLGA電紡絲膜進(jìn)行包裹,對(duì)照組只做單純縫合。分別在2、4、6周時(shí)間點(diǎn)各組處死5只兔取材,組織學(xué)和生物力學(xué)分析評(píng)估跟腱的愈合情況。 結(jié)果：PLGA電紡絲膜狀支架纖維直徑均勻,結(jié)構(gòu)規(guī)整,孔隙率82.6±3.45%；五種消毒方式均可達(dá)到滅菌目的；其中鉆6。照射和紫外線照射對(duì)支架結(jié)構(gòu)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(xué)特性明顯,體外增殖能力較強(qiáng),可以與PLGA電紡絲膜狀支架較好結(jié)合;MSCs組和PLGA電紡支架組組織學(xué)顯示肌腱愈合好,與周圍組織粘連較輕,生物力學(xué)優(yōu)于對(duì)照組。 結(jié)論：靜電紡絲技術(shù)是獲得納米級(jí)纖維的一種簡(jiǎn)便方法,使用梯度離心法分離獲得的MSCs純度較高,生物學(xué)特性明顯,體外增殖能力較強(qiáng),可以與PLGA電紡絲膜狀支架結(jié)合;MSCs附和PLGA電紡支架能夠促進(jìn)兔跟腱損傷的愈合,減少外源性疤痕組織長(zhǎng)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(xué)位授予單位】：中國(guó)人民解放軍軍醫(yī)進(jìn)修學(xué)院
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R318.08

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本文編號(hào)：1774043