Liu Pengcheng, Tan Qiuwen, Zhang Yi, Wang Hong, Lü Qing
Department of Breast Surgery, Clinical Research Center for Breast, West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China.
Research Core Facility of West China Hospital, Sichuan University, Chengdu Sichuan, 610041, P.R.China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2020 Oct 15;34(10):1322-1331. doi: 10.7507/1002-1892.202002126.
To investigate the effect of hydrogel from acellular porcine adipose tissue (HAPA) on the survival of transplanted adipose tissue.
For study, adipose tissue and HAPA-adipose tissue complex were cultured in normoxia and hypoxia atmospheres for 24 and 72 hours. TUNEL and Perilipin immunofluorescence staining were performed to observe the effect of HAPA on apoptosis and survival of adipocities. For study, 42 healthy male nude mice (4-6 weeks old) weighing 15-18 g were randomly divided into adipose group (group A), 10%HAPA group (group B), 20%HAPA group (group C), 30%HAPA group (group D), 40%HAPA group (group E), and 50%HAPA group (group F) according to different HAPA/adipose tissue volume ratio ( =7). For each group, 1 mL adipose tissue or HAPA-adipose tissue complex was injected subcutaneously into the dorsum of the nude mice. At 4 weeks after transplantation, 7 nude mice in each group were sacrificed and grafts were harvested, gross observation, volume measurement, ultrasound examination, and histologic staining (HE staining, CD31 and Perilipin immunofluorescence stainings) were applied.
Hypoxia showed a tendency of promoting adipose tissue necrosis and apoptosis, while HAPA exhibited an obvious effect of inhibiting cell apoptosis study ( <0.05). For study, grafts of all groups had intact fibrocapsule. No obvious signs of infection and necrosis were observed at 4 weeks. Volume shrinkage was observed in all groups, however, the groups A-D had significantly higher volume retention rate than groups E and F ( <0.05). Ultrasound examination showed that there were no significant difference in the number and volume of liquify area of the grafts in each group ( >0.05). With the increase of HAPA's volume ratio, HE staining proved an improved fat integrity while a gradually decreased vacuoles and fibrosis. CD31 immunohistochemical staining showed that the number of neo-vascularisation in groups E and F were significantly higher than those in groups A-D ( <0.05). Perilipin immunofluorescence staining showed that with the increase of HAPA volume ratio, the number of living adipocytes increased gradually, and more new adipocytes could be seen in the field of vision.
As the volume ratio of HAPA gradually increased, the survival of transplanted adipose tissue also increased, but the volume retention rate decreased gradually. 30%HAPA was considered the relative optimal volume ratio for its superior adipose tissue survival and volume retation rate.
探讨脱细胞猪脂肪组织水凝胶（HAPA）对移植脂肪组织存活的影响。
本研究中，脂肪组织和HAPA - 脂肪组织复合物在常氧和低氧环境中培养24小时和72小时。进行TUNEL和脂滴包被蛋白免疫荧光染色，以观察HAPA对脂肪细胞凋亡和存活的影响。本研究中，将42只体重15 - 18 g的健康雄性4 - 6周龄裸鼠，根据不同的HAPA/脂肪组织体积比（=7）随机分为脂肪组（A组）、10%HAPA组（B组）、20%HAPA组（C组）、30%HAPA组（D组）、40%HAPA组（E组）和50%HAPA组（F组）。每组将1 mL脂肪组织或HAPA - 脂肪组织复合物皮下注射到裸鼠背部。移植后4周，每组处死7只裸鼠并采集移植物，进行大体观察、体积测量、超声检查和组织学染色（HE染色、CD31和脂滴包被蛋白免疫荧光染色）。
低氧显示出促进脂肪组织坏死和凋亡的趋势，而HAPA在本研究中表现出明显的抑制细胞凋亡作用（<0.05）。本研究中，所有组的移植物均有完整的纤维包膜。4周时未观察到明显的感染和坏死迹象。所有组均观察到体积缩小，然而，A - D组的体积保留率显著高于E组和F组（<0.05）。超声检查显示，每组移植物液化区的数量和体积无显著差异（>0.05）。随着HAPA体积比的增加，HE染色显示脂肪完整性改善，而空泡和纤维化逐渐减少。CD31免疫组织化学染色显示，E组和F组的新生血管数量显著高于A - D组（<0.05）。脂滴包被蛋白免疫荧光染色显示，随着HAPA体积比的增加，存活脂肪细胞数量逐渐增加，视野中可见更多新的脂肪细胞。
随着HAPA体积比逐渐增加，移植脂肪组织的存活率也增加，但体积保留率逐渐降低。30%HAPA因其优异的脂肪组织存活率和体积保留率被认为是相对最佳的体积比。
