Abstract：
BACKGROUND: Autologous fat transplantation is limited by the uncertainty of graft retention, impeding its application. Among the current strategies for processing lipoaspirates, high-density fat (HDF) is recommended owing to the enrichment of stem cells and washing before cotton concentration for simplicity of operation. Poloxamer 188 (P188) washing has been shown to repair the membranes of damaged cells. This study aimed to investigate the effect of P188-washing on fat graft survival and identify the best technique for processing lipoaspirates.
METHODS: Lipoaspirates were prepared using centrifugation to obtain HDF, which was then washed with saline or P188 followed by cotton concentration. Tissue integrity, adipocytic activity, and viability of stromal vascular fraction (SVF) in the samples from the 3 groups were assessed. Samples were sequenced in vitro using high-throughput RNA-seq, and differentially expressed genes were validated using qPCR and western blotting (WB). After transplantation under the dorsum of nude mice for 8 weeks, the grafts were extracted and examined for residual volume, histologic characteristics, and vascularization.
RESULTS: The HDF and P188 groups showed a higher survival rate of SVF, more Ki67-positive cells, intact tissue structure, and lesser fibrosis than the saline group. There were no significant differences in the density of SVF and residual volume of grafts. HDF showed significantly improved vascularization during 8 weeks. Through RNA-seq and bioinformatic analysis, notable changes in several related genes after transplantation were observed.
CONCLUSIONS: P188 treatment can prevent cells from apoptosis and preserve tissue viability, thereby improving graft quality. HDF contains large amounts of SVF and can be regarded as an excellent grafting material.
METHODS: Lipoaspirates were prepared using centrifugation to obtain HDF, which was then washed with saline or P188 followed by cotton concentration. Tissue integrity, adipocytic activity, and viability of stromal vascular fraction (SVF) in the samples from the 3 groups were assessed. Samples were sequenced in vitro using high-throughput RNA-seq, and differentially expressed genes were validated using qPCR and western blotting (WB). After transplantation under the dorsum of nude mice for 8 weeks, the grafts were extracted and examined for residual volume, histologic characteristics, and vascularization.
RESULTS: The HDF and P188 groups showed a higher survival rate of SVF, more Ki67-positive cells, intact tissue structure, and lesser fibrosis than the saline group. There were no significant differences in the density of SVF and residual volume of grafts. HDF showed significantly improved vascularization during 8 weeks. Through RNA-seq and bioinformatic analysis, notable changes in several related genes after transplantation were observed.
CONCLUSIONS: P188 treatment can prevent cells from apoptosis and preserve tissue viability, thereby improving graft quality. HDF contains large amounts of SVF and can be regarded as an excellent grafting material.
摘要：
背景：自体脂肪移植受到移植物保留不确定性的限制，阻碍其应用。在目前处理脂肪抽吸物的策略中，建议使用高密度脂肪（HDF），因为为了简化操作，在浓缩棉花前进行干细胞和洗涤。已经显示泊洛沙姆188(P188)洗涤修复受损细胞的膜。本研究旨在研究P188洗涤对脂肪移植物存活的影响，并确定处理脂肪抽吸物的最佳技术。
方法：使用离心制备脂肪抽吸物，以获得HDF，然后用盐水或P188洗涤，然后浓缩棉花。组织完整性,脂肪细胞活动，评估3组样品中基质血管分数（SVF）的活力。使用高通量RNA-seq对样品进行体外测序，差异表达的基因使用qPCR和蛋白质印迹（WB）进行验证。裸鼠背部移植8周后，提取移植物并检查剩余体积，组织学特征，和血管化。
结果：HDF组和P188组表现出更高的SVF存活率，更多的Ki67阳性细胞，完整的组织结构，纤维化程度低于生理盐水组。SVF的密度和移植物的残余体积没有显着差异。HDF在8周内显示出显著改善的血管形成。通过RNA-seq和生物信息学分析，移植后观察到几个相关基因的显著变化。
结论：P188处理可以防止细胞凋亡和保持组织活力，从而提高嫁接质量。HDF中含有大量的SVF,是一种优良的接枝材料。
方法：使用离心制备脂肪抽吸物，以获得HDF，然后用盐水或P188洗涤，然后浓缩棉花。组织完整性,脂肪细胞活动，评估3组样品中基质血管分数（SVF）的活力。使用高通量RNA-seq对样品进行体外测序，差异表达的基因使用qPCR和蛋白质印迹（WB）进行验证。裸鼠背部移植8周后，提取移植物并检查剩余体积，组织学特征，和血管化。
结果：HDF组和P188组表现出更高的SVF存活率，更多的Ki67阳性细胞，完整的组织结构，纤维化程度低于生理盐水组。SVF的密度和移植物的残余体积没有显着差异。HDF在8周内显示出显著改善的血管形成。通过RNA-seq和生物信息学分析，移植后观察到几个相关基因的显著变化。
结论：P188处理可以防止细胞凋亡和保持组织活力，从而提高嫁接质量。HDF中含有大量的SVF,是一种优良的接枝材料。
