Variables affecting the retained volume of adipose tissue after transplantation in rats

doi:10.3969/j.issn.2095-4344.1486

Abstract

Abstract:

BACKGROUND: The high and unstable absorption rate after autologous granule fat transplantation limits its clinical application. In addition to the early establishment of blood supply, it is particularly important to find new ways to improve survival rate.
OBJECTIVE: To investigate the effect of mechanical shearing and anti-apoptotic drug edaravone on the volume retention after autologous fat transplantation from the perspective of apoptosis.
METHODS: The study was approved by the Laboratory Animal Ethics Committee of Anhui Medical University. Forty-two Sprague-Dawley male rats were randomly divided into three groups: A, B, and C. The rats were transplanted with fat by clinical autologous fat transplantation. Group A was the control group (n=6): the adipose tissue was given 130 times of basic shearing during operation; the B and C groups were given different variables of adipose tissue on the basis of group A. Group B was a mechanical shearing group with a total of 18 rats subdivided into subgroups B1, B2 and B3, which were given 230 times, 330 times and 430 mechanical shears, respectively. Group C was the edaravone group (n=18), subdivided into C1, C2 and C3 subgroups, respectively, given 0.1%, 0.2% and 1% concentration of edaravone, and then the adipose tissue was given 130 times of basic shearing. Graft specimens were taken immediately after operation and at 1, 4 and 8 weeks after transplantation, to observe and record the morphological features and volume of the specimens. The histological changes were observed under a light microscope after hematoxylin-eosin staining. The apoptosis ratio of fat cells in each group was detected by flow cytometry Annexin V/PI double staining.
RESULTS AND CONCLUSION: (1) Compared with the control group, the group B: the proportion of apoptosis was increased significantly at immediately and 1 week after surgery, and the graft absorption rate was fast; at 4 and 8 weeks after operation, the volume of hematoxylin- eosin-stained adipocytes was different, the arrangement was disordered, and the graft retention volume was reduced. The graft retention volume was reduced (P < 0.05). C1 and C2 subgroups: the proportion of apoptosis in specimens was significantly decreased at immediately and 1 week after surgery. At 4 and 8 weeks after operation, the morphology of hematoxylin-eosin-stained adipocytes was more regular and volume was more consistent, more blood vessels were visible. The graft retention volume was significantly increased (P < 0.05). (2) There was no significant difference in the fat absorption rate between C3 subgroup and control group (P > 0.05). (3) To conclude, in autologous fat transplantation, mechanical shearing reduces the volume of fat retained after transplantation, and a certain concentration of anti-apoptotic drug edaravone can increase the retained volume of adipose tissue after transplantation.

Key words: fat cells, autograft, apoptosis, edaravone, adipose tissue, absorption of autograft

CLC Number:

Jia Daping, Ma Li, Wu Liangliang, Zhao Yu. Variables affecting the retained volume of adipose tissue after transplantation in rats[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(35): 5652-5657.

Figures/Tables 4

表2结果表明，随时间延长，大鼠移植后脂肪体积不断缩小，对同期所有实验亚组与对照组比较，7个亚组间数据具有方差齐性，行单因素方差分析，7个组间移植物留存体积不完全相等(F1周=45.768、F4周=122.049、F8周=59.245，P < 0.05)，其中B组移植物留存体积较对照组小，C1和C2移植物留存体积显著大于对照组(P < 0.05)。对同一时期B组、C组内亚组间两两对比留存体积，行SNK检验，1周时，B3对比B1、B2组移植物留存体积明显较小(FB组组间= 16.422，P < 0.05)，C组各亚组移植物留存体积差异无显著性意义(FC组组间=3.256，P > 0.05)；4周时，B组各亚组移植物留存体积成梯度下降，B3<B2<B1，C1、C2对比C3组移植物留存体积显著增加(FB组组间=18.514，FC组组间= 35.975，P < 0.05)；8周时B3与B1留存体积比较明显缩小(FB组组间=4.339，P < 0.05)，C组各亚组移植物留存体积从大到小排序为C2>C1>C3，差异有显著性意义(FC组组间= 44.322，P < 0.05)。以上统计结果表明，大鼠自体脂肪组织移植中给予获取的脂肪浓度不超过0.2%的依达拉奉注射能明显提高移植后脂肪组织体积留存率，机械剪切会显著降低脂肪组织移植后体积留存。表3结果显示，8周时，相比对照组，B3组脂肪组织体积吸收率增加了约14%，C2组脂肪组织体积吸收率降低约16%，C3组移植后脂肪吸收率与同期对照组比较差异无显著性意义(P > 0.05)。 2.4 组织学比较苏木精-伊红染色结果观察分析发现：术后1，4周时，各组脂肪细胞形态均维持较好，呈圆形或规则多边形，细胞壁完整，细胞核偏向一侧，细胞间间隙稳定，可见明显炎症细胞浸润；术后8周，对照组可见脂肪细胞体积不一，结构紊乱、毛细血管增生，纤维组织包饶脂肪细胞，B3组脂肪细胞排列紊乱，间质有纤维组织增生、炎性细胞浸润，可见脂肪细胞形态明显不规则，见较多脂肪空泡形成，C2组脂肪细胞轮廓清晰，细胞形态较规则，细胞体积较一致，可见丰富毛细血管增生，仅有少量脂肪空泡形成。B组、C组典型图片见图2。 "

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