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首页> 外文期刊>European Cells & Materials >Pre-cultivation of adipose tissue-derived microvascular fragments in porous scaffolds does not improve their in vivo vascularisation potential
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Pre-cultivation of adipose tissue-derived microvascular fragments in porous scaffolds does not improve their in vivo vascularisation potential

机译：多孔支架中脂肪组织来源的微血管片段的预培养不能提高其体内血管化潜力

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Adipose tissue-derived microvascular fragments represent promising vascularisation units for implanted tissue constructs. However, their reassembly into functional microvascular networks takes several days, during which the cells inside the implants are exposed to hypoxia. In the present study, we analysed whether this critical phase may be overcome by pre-cultivation of fragment-seeded scaffolds prior to their implantation. Green fluorescent protein (GFP)-positive microvascular fragments were isolated from epididymal fat pads of male C57BL/6-TgN (ACTB-EGFP) 1Osb/J mice. Nano-size hydroxyapatite particles/poly (ester-urethane) scaffolds were seeded with these fragments and cultivated for 28 days. Subsequently, these scaffolds or control scaffolds, which were freshly seeded with GFP-positive microvascular fragments, were implanted into the dorsal skinfold chamber of C57BL/6 wild-type mice to study their vascularisation and incorporation by means of intravital fluorescence microscopy, histology and immunohistochemistry over 2 weeks. Pre-cultivation of microvascular fragments resulted in the loss of their native vessel morphology. Accordingly, pre-cultivated scaffolds contained a network of individual CD31/GFP-positive endothelial cells with filigrane cell protuberances. After implantation into the dorsal skinfold chamber, these scaffolds exhibited an impaired vascularisation, as indicated by a significantly reduced functional microvessel density and lower fraction of GFP-positive microvessels in their centre when compared to freshly seeded control implants. This was associated with a deteriorated incorporation into the surrounding host tissue. These findings indicate that freshly isolated, non-cultivated microvascular fragments should be preferred as vascularisation units. This would also facilitate their use in clinical practice during intra-operative one-step procedures.
机译：源自脂肪组织的微血管片段代表了用于植入组织构建体的有希望的血管形成单位。然而，它们重新组装成功能性微血管网络需要几天，在此期间，植入物内部的细胞会暴露于缺氧状态。在本研究中，我们分析了是否可以通过在种子植入前对片段播种的支架进行预培养来克服这一关键阶段。从雄性C57BL / 6-TgN（ACTB-EGFP）1Osb / J小鼠的附睾脂肪垫中分离出绿色荧光蛋白（GFP）阳性微血管片段。用这些片段播种纳米级羟基磷灰石颗粒/聚（酯-氨基甲酸酯）支架，并培养28天。随后，将新鲜接种有GFP阳性微血管片段的这些支架或对照支架植入C57BL / 6野生型小鼠的背侧皮腔，通过活体荧光显微镜，组织学和免疫组化研究其血管化和整合超过2周。微血管片段的预培养导致其天然血管形态的丧失。因此，预先培养的支架包含具有纤毛细胞突起的单个CD31 / GFP阳性内皮细胞的网络。与新鲜植入的对照植入物相比，植入到背部皮褶室后，这些支架显示出血管化受损，这是由功能性微血管密度显着降低和中心的GFP阳性微血管分数降低所表明的。这与并入周围宿主组织中的变质有关。这些发现表明，新鲜分离的，未培养的微血管片段应优选作为血管生成单位。这也将有助于它们在术中一步操作过程中在临床实践中的使用。

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