From the Division of Plastic and Reconstructive Surgery, Department of Surgery, Taipei Veterans General Hospital.
Ann Plast Surg. 2022 Mar 1;88(1s Suppl 1):S4-S12. doi: 10.1097/SAP.0000000000003075.
BACKGROUND: Fresh fat grafts are commonly used in both esthetic and reconstructive surgeries, but the graft resorption rate varies. Cryopreservation of unused fat for later touch-up is one option to resolve this variation. In our previous studies, we found that fat cryopreservation may be a practical strategy for storing fat tissue. To explore the cryopreservation method, we evaluated the role of vascular endothelial growth factor (VEGF) in human frozen fat grafts. METHODS: The concentration of VEGF in human frozen fat grafts subjected to different preservation times was determined using Western blotting and enzyme-linked immunosorbent assay. The angiogenic effect of frozen fat grafts was evaluated using a chorioallantoic membrane assay. Furthermore, the impact of adding human adipose-derived stem cells (hADSCs) or different concentrations of avastin (bevacizumab) to frozen fat grafts on angiogenesis was assessed. The viability of frozen fat grafts with or without hADSCs was evaluated using a nude mouse implantation study. Explanted fat tissues were examined on days 1, 4, 7, 14, 28, and 90, and morphological and histological analyses, immunohistochemistry, and enzyme-linked immunosorbent assay (VEGF concentration) were carried out. RESULTS: No significant difference in VEGF concentration between fresh and frozen fat was observed with respect to preservation duration. In the chorioallantoic membrane assay, frozen fat grafts with hADSCs displayed significantly enhanced angiogenesis. Avastin was found to decrease angiogenesis in frozen fat grafts. However, in the nude mouse implantation study, frozen fat grafts displayed VEGF maintenance, with the highest concentration observed on day 7. Adding hADSCs to the graft further increased the VEGF concentration and CD31 expression. Fat graft viability was found to be higher in the frozen fat grafts containing hADSCs than in grafts without hADSCs. CONCLUSIONS: Human fat grafts can maintain VEGF expression under frozen conditions for at least 12 months. The addition of hADSCs to the frozen fat graft could further enhance angiogenesis, VEGF expression, and fat cell viability.
背景：新鲜脂肪移植物常用于美容和重建手术，但移植物吸收率存在差异。冷冻保存未使用的脂肪以备后续补注是解决这种变化的一种选择。在我们之前的研究中，我们发现脂肪冷冻保存可能是储存脂肪组织的一种实用策略。为了探索冷冻保存方法，我们评估了血管内皮生长因子（VEGF）在人冷冻脂肪移植物中的作用。
方法：使用 Western blot 和酶联免疫吸附试验测定不同保存时间后人冷冻脂肪移植物中 VEGF 的浓度。通过鸡胚尿囊膜试验评估冷冻脂肪移植物的血管生成作用。此外，评估向冷冻脂肪移植物中添加人脂肪源性干细胞（hADSCs）或不同浓度阿瓦斯汀（贝伐单抗）对血管生成的影响。使用裸鼠植入研究评估有无 hADSCs 的冷冻脂肪移植物的存活率。在第 1、4、7、14、28 和 90 天，对植入的脂肪组织进行检查，进行形态学和组织学分析、免疫组织化学和酶联免疫吸附试验（VEGF 浓度）。
结果：新鲜和冷冻脂肪的 VEGF 浓度在保存时间方面没有显著差异。在鸡胚尿囊膜试验中，含有 hADSCs 的冷冻脂肪移植物显示出明显增强的血管生成作用。阿瓦斯汀被发现降低了冷冻脂肪移植物中的血管生成。然而，在裸鼠植入研究中，冷冻脂肪移植物显示出 VEGF 的维持，第 7 天观察到最高浓度。向移植物中添加 hADSCs 进一步增加了 VEGF 浓度和 CD31 表达。在含有 hADSCs 的冷冻脂肪移植物中，脂肪移植物的存活率高于不含 hADSCs 的移植物。
结论：人脂肪移植物在冷冻条件下至少可维持 12 个月的 VEGF 表达。向冷冻脂肪移植物中添加 hADSCs 可以进一步增强血管生成、VEGF 表达和脂肪细胞存活率。
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