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聚乳酸-乙醇酸共聚物编织网/胶原-壳聚糖混合支架的血管化

[Vascularization of poly-lactic-co-glycolic acid knitted mesh/collagen-chitosan hybrid scaffold].

作者信息

Wang Xin-gang, Han Chun-mao, You Chuan-gang, Zheng Yu-rong, Li Qi-yin, Zhang Li-ping

机构信息

Department of Burns, the Second Affiliated Hospital of Zhejiang University College of Medicine, Hangzhou 310009, China.

出版信息

Zhonghua Shao Shang Za Zhi. 2011 Aug;27(4):260-4.

Abstract

OBJECTIVE

To investigate the effects of poly-lactic-co-glycolic acid (PLGA) knitted mesh/collagen-chitosan hybrid scaffold (PCCS) on angiogenesis, and to explore the relative mechanisms.

METHODS

PLGA knitted mesh was integrated into collagen-chitosan scaffold (CCS) to construct PCCS with freeze-lyophilizing method, and CCS was made with the same method. The characteristics of morphology and water absorbing capacity among PCCS, PLGA knitted mesh, and CCS were compared in vitro. PCCS and CCS was respectively implanted into subcutaneous tissue of back on both sides in 24 SD rats, and the tissue specimens were harvested at post operation week (POW) 1, 2, and 4 according to the random number table to evaluate the level of angiogenesis by histopathological and immunohistochemical examinations. The expression levels of alpha smooth muscle actin (alpha-SMA) and vascular endothelial growth factor (VEGF) mRNA were examined by real-time quantitative RT-PCR. Data were processed with t test.

RESULTS

(1) PLGA knitted mesh was closely integrated with sponge of collagen-chitosan in PCCS, and the porous structure of PCCS was similar to that of CCS. (2) Compared with that of PCCS [(506 +/- 15)%], the water absorbing rate of CCS and PLGA knitted mesh was respectively increased and decreased [(627 +/- 21)%, (195 +/- 15)%, with t value respectively 3.8, 11.9, P < 0.05 or P < 0.001]. (3) The scaffolds were filled with newly formed tissue in CCS at POW 4, while those in PCCS were observed at POW 2 with more homogeneous and abundant collagen. (4) Blood vessels could be induced, and they grew into scaffolds along with prolongation of implantation time in PCCS and CCS. The number of mature blood vessels in PCCS at POW 1, 2, 4 [(10.7 +/- 3.2), (18.6 +/- 2.1), and (30.3 +/- 4.5) branches per square centimeter] was respectively higher than that in CCS [(5.4 +/- 0.9), (10.8 +/- 4.2), and (23.6 +/- 1.7) branches per square centimeter, with t value respectively 4.6, 4.4, 4.5, P values all below 0.01]. (5) The expression levels of alpha-SMA and VEGF mRNA in PCCS at POW 1, 2, 4 were significantly higher than those in the CCS (with t(alpha-SMA) value respectively 1.26, 1.63, 2.17, with t(VEGF) value respectively 5.52, 2.07, 1.78, P values all below 0.01).

CONCLUSIONS

PCCS is able to induce the ingrowth of blood vessels rapidly and promote their maturity. The mechanical properties and microstructures of scaffolds play synergistic role in the process of angiogenesis.

摘要

目的

研究聚乳酸-乙醇酸共聚物(PLGA)编织网/胶原-壳聚糖混合支架(PCCS)对血管生成的影响,并探讨其相关机制。

方法

采用冷冻干燥法将PLGA编织网与胶原-壳聚糖支架(CCS)复合构建PCCS,同时用相同方法制备CCS。体外比较PCCS、PLGA编织网及CCS的形态特征和吸水能力。将PCCS和CCS分别植入24只SD大鼠背部两侧皮下组织,根据随机数字表于术后第1、2、4周取材,通过组织病理学和免疫组织化学检查评估血管生成水平。采用实时定量RT-PCR检测α平滑肌肌动蛋白(α-SMA)和血管内皮生长因子(VEGF)mRNA的表达水平。数据采用t检验处理。

结果

(1)PCCS中PLGA编织网与胶原-壳聚糖海绵紧密结合,PCCS的多孔结构与CCS相似。(2)与PCCS的吸水率[(506±15)%]相比,CCS和PLGA编织网的吸水率分别升高和降低[(627±21)%,(195±15)%,t值分别为3.8、11.9,P<0.05或P<0.001]。(3)术后4周时CCS中有新形成的组织填充支架,而PCCS在术后2周时即观察到有更均匀且丰富的胶原。(4)PCCS和CCS均可诱导血管生成,且随着植入时间延长血管长入支架。术后第1、2、4周时PCCS中成熟血管数量[每平方厘米(10.7±3.2)、(18.6±2.1)和(30.3±4.5)支]分别高于CCS中的[每平方厘米(5.4±0.9)、(10.8±4.2)和(23.6±1.7)支,t值分别为4.6、4.4、4.5,P值均<0.01]。(5)术后第1、2、4周时PCCS中α-SMA和VEGF mRNA的表达水平显著高于CCS(α-SMA的t值分别为1.26、1.63、2.17,VEGF的t值分别为5.52、2.07、1.78,P值均<0.01)。

结论

PCCS能够快速诱导血管长入并促进其成熟。支架的力学性能和微观结构在血管生成过程中起协同作用。

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