机构地区:[1]南方医科大学南方医院整形外科,广州510515
出 处:《中国修复重建外科杂志》2014年第5期644-648,共5页Chinese Journal of Reparative and Reconstructive Surgery
基 金:国家自然科学基金资助项目(81171834;81372083);广东省自然科学基金资助项目(S201 1010003058;S2012010 009370);广东省科技计划项目(2 011B061300015);广州市科技计划重点项目(11C32120716);教育部博士点专项基金(20114433120007)~~
摘 要:目的 初步探讨兔脂肪组织来源血管基质片段(stromal vascular fraction,SVF)促进组织工程室内再血管化和诱导组织再生的机制。 方法 取6月龄新西兰大白兔24只,雌雄不限,体重2.5~2.8 kg;将Ⅰ型胶原支架与胸背动静脉束复合移植于背侧,外置圆柱形中空硅胶小室,并随机分为两组(n=12)。术后2周,实验组取颈背部脂肪垫按照胶原酶消化法分离SVF,DiI标记后将1 mL SVF细胞悬液(1 × 106个/mL)注入圆柱形中空硅胶小室内;对照组注射等量生理盐水。注射后2、4周取出新生组织行大体、组织学观察,4周时实验组行免疫荧光染色鉴定。 结果 注射后2周两组新生组织呈圆柱形,表面可见明显血管网络分布及未完全降解的Ⅰ型胶原支架,实验组新生组织体积为(0.87 ± 0.11) mL,大于对照组的(0.72 ± 0.08)mL(t=2.701,P=0.011);4周时实验组未见支架残留,对照组可见有少许絮状支架残留,新生组织体积均较2周时缩小,分别为(0.74 ± 0.14)、(0.64 ± 0.10)mL,组间比较差异无统计学意义(t=1.424,P=0.093)。组织学观察示,注射后两组均有新生血管,且4周血管基本成熟,但无脂肪团或脂肪小叶形成;2、4周时实验组新生血管数均显著多于对照组(t=6.291,P=0.000;t=5.445,P=0.000)。注射后4周荧光显微镜下观察示,实验组SVF在新生组织中存活并主要集中在新生组织边缘,部分血管内皮细胞同时表达CD31和DiI双阳性。 结论 兔SVF能促进组织工程室内新生组织的血管化并增加组织体积,但是在缺乏成脂微环境诱导下不能自主分化为脂肪细胞。Objective To evaluate the mechanism of stromal vascular fraction (SVF) promoting angiogenesis and tissue regeneration in tissue engineering chamber. Methods Twenty-four 6-month-old New Zealand white rabbits, male or female, weighing 2.5-2.8 kg, were selected. Thoracic dorsal arteriovenous bundle combined with collagen type I scaffold was transplanted to dorsal side, and wrapped by cylindrical hollow silicone chamber; all animals were randomly divided into the experimental group (n=12) and the control group (n=12). SVF was isolated from the back fat pads of rabbits in experimental group and labelled with DiI at 2 weeks after operation. The 1 mL cell suspension (1 × 106 cells/mL) and equal saline were injected into the chamber in experimental group and control group, respectively. The regenerative tissues were harvested for general observation and HE staining at 2 and 4 weeks after injection;and immunofluorescent staining was carried out in experimental group at 4 weeks. Results At 2 weeks after injection, the regenerative tissue was cylindrical; obvious vessel network and incompletely degradable collagen scaffold could be seen on the surface of the new tissue in 2 groups. The volume of new tissue was (0.87 ± 0.11) mL in experimental group, and (0.72 ± 0.08) mL in control group at 2 weeks, showing significant difference (t=2.701, P=0.011). At 4 weeks, little collagen scaffold could be seen on the surface in control group, but no collagen scaffold in experimental group; the volume of new tissue was (0.74 ± 0.14) mL in experimental group, and (0.64 ± 0.10) mL in control group, showing no significant difference (t=1.424, P=0.093). HE staining showed new mature vessels at 4 weeks, but no adipose tissue or fat lobulus formed in both groups; the capillary density was significantly higher in experimental group than in control group at 2 weeks (t=6.291, P=0.000) and at 4 weeks (t=5.445, P=0.000). The immunofluorescent staining found that SVF survived and located at the e
关 键 词:脂肪组织工程 组织工程室 血管基质片段 血管化 兔
分 类 号:R318[医药卫生—生物医学工程] R622[医药卫生—基础医学]
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