Human preadipocytes seeded on freeze-dried collagen scaffolds investigated in vitro and in vivo
Human preadipocytes seeded on freeze-dried collagen scaffolds investigated in vitro and in vivo
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DOI：
10.1016/s0142-9612(00)00186-1
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发表时间：
2001-03-01
期刊：
影响因子：
14
通讯作者：
Pallua, N
中科院分区：
文献类型：
--
作者：
von Heimburg, D;Zachariah, S;Pallua, N
目前，没有足够的植入材料用于矫正软组织缺陷，例如大面积深度烧伤后、肿瘤切除后以及遗传性和先天性缺陷（例如Romberg病、波兰综合征）。成熟脂肪组织的自体移植效果不佳。本研究分离培养了青年人前脂肪细胞。将10（6）个前脂肪细胞接种到具有均匀40 μ m纯尺寸和规则层状结构的胶原海绵上，并植入免疫缺陷小鼠中。在对照中使用不含前脂肪细胞的胶原海绵。在体外培养24 h后以及移植后3周和8周，对大体印象、重量、厚度、组织学、免疫组织化学（支架结构、细胞构成、接种细胞的穿透深度）和超微结构进行评估。前脂肪细胞在植入前以299 +/- 55 μ m的深度接种后24小时穿透支架。在3周和8周后，在所有前脂肪细胞/胶原移植物上肉眼观察到伴有新血管的体内脂肪组织层。对照移植物无变化，无血管长入。在体外24小时和体内3周之间，所有移植物的重量显著减轻（p < 0.05），而从第3周至第8周仅轻微减轻。所有移植物的厚度在前3周内均减小（p < 0.05）。前脂肪细胞/胶原移植物较薄，但在该时间点具有比对照更高的重量。组织学显示脂肪组织和丰富的血管形成粘附在胶囊下的支架上。对照海绵仅含有少量细胞和囊，但没有脂肪组织。在所有前脂肪细胞胶原移植物中发现人波形蛋白阳性细胞，但在对照组中没有，穿透1188 +/- 498 μ m（3周）和1433 +/- 685 μ m（8周）。超微结构分析表明，完整的活的脂肪细胞在海绵与前脂肪细胞接种体内分化。细胞外基质的形成在前脂肪细胞/胶原移植物中更为明显。将分离培养的前脂肪细胞移植到标准化的胶原基质中，可获得血管化良好的脂肪样组织。假设需要大于40 μ m的孔径，因为前脂肪细胞在分化过程中由于脂质的掺入而扩大。(C)2001爱思唯尔科技有限公司版权所有。
Currently, there is no adequate implant material for the correction of soft tissue defects such as after extensive deep burns, after tumor resection and in hereditary and congenital defects (e.g. Romberg's disease, Poland syndrome). The autologous transplantation of mature adipose tissue has poor results. In this study human preadipocytes of young adults were isolated and cultured. 10(6) preadipocytes were seeded onto collagen sponges with uniform 40 mum pure size and regular lamellar structure and implanted into immunodeficient mice. Collagen sponges without preadipocytes were used in the controls. Macroscopical impression, weight, thickness, histology, immunohistochemistry (scaffold structure, cellularity, penetration depth of the seeded cells) and ultrastructure were assessed after 24 h in vitro and after explantation at 3 and 8 weeks. Preadipocytes penetrated the scaffolds 24 h after seeding at a depth of 299 +/- 55 mum before implantation. Macroscopically after 3 and 8 weeks in vivo layers of adipose tissue accompanied by new vessels were found on all preadipocyte/collagen grafts. The control grafts appeared unchanged without vessel ingrowth. There was a significant weight loss of all grafts between 24 h in vitro and 3 weeks in vivo (p < 0.05), whereas there was only a slight weight reduction from week 3 to 8. The thickness decreased in the first 3 weeks (p < 0.05) in all grafts. The preadipocyte/collagen grafts were thinner but had a higher weight than the controls at this point in time. The histology showed adipose tissue and a rich vascularisation adherent to the scaffolds under a capsule. The control sponges contained only few cells and a capsule but no adipose tissue. Human-vimentin positive cells were found in all preadipocyte collagen grafts but not in the controls, penetrating 1188 +/- 498 mum (3 weeks) and 1433 +/- 685 mum (8 weeks). Ultrastructural analysis showed complete in vivo differentiation of viable adipocytes in the sponge seeded with preadipocytes. Formation of extracellular matrix was more pronounced in the preadipocyte/collagen grafts. The transplantation of isolated and cultured preadipocytes within a standardised collagen matrix resulted in well-vascularised adipose-like tissue. It is assumed that a pore size greater than 40 mum is required, as preadipocytes enlarge during differentiation due to incorporation of lipids. (C) 2001 Elsevier Science Ltd. All rights reserved.