| Objective1. To explore the effect of different processing methods on lipoaspirate by examining the cell morphology, aqueous fluid, free lipid content, the number of stromal vascular fraction (SVF) cells, adipose-derived stem cells (ADSC) and blood-derived cells, the activity of glycerol 3-phosphate dehydrogenase (G3PDH) of the fat grafts;2. The fat grafts prepared by different methods were transplanted subcutaneously into nude mice. To evaluate the effects on survival rate of transplanted fat which prepared by different processing methods were measured by examining the weight and histological characteristics of the new tissues.3. To evaluate the methods of treating micromastia and breast defect after mastectomy with the fat grafts which were processed by cotton pad concentration.Methods1. Measure the indicators of fat grafts which were prepared by different processing methods in four groups by four different methods as gravity sedimentation, washing and gravity sedimentation, centrifugation, and cotton pad concentration. The integrity of the fat cells, aqueous fluid and free lipid content, number of SVF cells, ADSC and blood-derived cells, and the activity of G3PDH were observed.2. Examining the results of fat grafts by different processing methods in nude mice of four groups with different methods including gravity sedimentation, washing and gravity sedimentation, centrifugation and cotton pad concentration, each group 15 mice. Fat grafting was performed for three times, with adipose tissue from a patient each time. The approach of fat harvesting, grouping and processing is the same as mentioned in the first part. The weight of the new tissue was measured and the histological characteristics were evaluated.3. Treating micromastia and breast defect after mastectomy with the fat grafts which were processed by cotton pad concentration. The results were measured by comparing the preoperative and postoperative photos of the patients, follow-up postoperative satisfaction, the change of chest circumference between preoperation and postoperation, the breast volume change of patients by D Camera (axisthree) examination, and the detection of breast cysts and lumps by physical examination and MRIResultsPart One(1) HE and immunohistochemical staining showed:the integrity of the adipocytes is not significantly different among the four groups with different processing methods as gravity sedimentation, washing and gravity sedimentation, centrifugation and cotton pad concentration.(2) The number of aqueous fluid in group of gravity sedimentation, washing and gravity sedimentation is more than in group of centrifugation and cotton pad concentration (p<0.01). The free lipid content in group of cotton pad concentration is the lowest (p<0.05); the free lipid content in group of centrifugation is less than in group of gravity sedimentation, and group of washing and gravity sedimentation (P<0.05).(3) The number of SVF cells in group of gravity sedimentation is less than group of centrifugation and cotton pad concentration. There are no significant differences in other groups.(4) Flow cytometric analysis:the number of CD45 (+) cells in group of gravity sedimentation is more than those in group of cotton pad concentration (P<0.05), the number of CD31 (+) CD45 (-) cells in group of washing and gravity sedimentation is more than those in group of gravity sedimentation (P<0.05), There are no significant differences between other groups.(5) The activity of G3PDH in group of gravity sedimentation, washing and gravity sedimentation is less than that in group centrifugation and cotton pad concentration. There are no significant differences between other groups.Part Two(1) The weight of the newborn tissue in group of cotton pad concentration (0.149 ± 0.068g)is more than that in group of gravity sedimentation(0.081 ± 0.041g), and group of washing and gravity sedimentation(0.093 ± 0.060g) and group of centrifugation (0.085 ± 0.053g) (p<0.05). There are no significant differences among other groups.(2) HE staining shows that the presence of intact and nucleated fat cells, cysts and vacuoles formation, fibrosis, infiltration of lymphocytes and macrophages were of no significant difference.(3) Perilipin immunohistochemical staining shows that the cells with perilipin expression in group of cotton pad concentration are more than in those in other groups, there are no significant differences among the other groups.Part Three(1) Treating micromastia with autologous fat grafting which were processed by cotton pad concentration. The results showed that the breast is natural and soft, the satisfaction of patients is high, the change of chest circumference is significant, and the rate of fat survival is about 44.4±6.6%.(2) Breast reconstruction after mastectomy in breast cancer is done with autologous fat grafting which were processed by cotton pad concentration. The results showed that the surgical trauma is limited, and the postoperative patient satisfaction is high.Conclusion1. In vitro experiments showed that the indicators in group of centrifugation and cotton pad concentration were better than in group gravity sedimentation, washing and gravity sedimentation. Especially the aqueous fluid and free lipid of adipose tissue in group of centrifugation and cotton pad concentration were significantly less than those in group of gravity sedimentation, washing and gravity sedimentation.In animal experiments, the results showed that the survival rate of adipose tissue in group of gravity sedimentation, washing and gravity sedimentation, centrifugation was not significantly different, but the survival rate of adipose tissue in group of cotton pad concentration was significantly higher than that in the other three groups. Therefore, cotton pad concentration is a better fat concentration method.2. Autologous fat particle is a good soft tissue filler. Treat micromastia and breast defect after mastectomy with processed fat grafts, we can obtain satisfactory clinical results. The fat processing method by cotton pad concentration warrant further evaluation.How to improve the survival rate of adipose tissue after autologous fat transplantation has become the key point in the fat transplantation field. The survival rate is closely related to the fat processing methods. Adipose tissue after processing should contain water, grease particles, blood-derived cells and cell debris as little as possible, and maintain the integrity of the fat cells as far as possible, and adipose-derived stem cells as rich as possible. Currently, the processing methods of fat tissue mainly include gravity sedimentation, washing and gravity sedimentation, centrifugation, filtration and concentration. The aqueous fluid of fat tissue is about 30% in processing with gravity sedimentation, washing and gravity sedimentation. Autologous fat grafting in breast augmentation needs a large amount of fat tissue. The process of centrifugation is complex, time-consuming, increased possibility of surgical infection while processing off the operating table. There is a closed system to filter and concentrate the tat tissue abroad, however, the device is expensive with high financial burden on patients. In our clinical study, the method is simple and efficient by cotton pad concentration on adipose tissue. The operation is on the operating table, avoiding the possibility of infection. In our clinical study, the result is good, with high satisfaction and few complications. No clinical infections were observed. |
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