肥胖患者减重成功后其脂肪干细胞的生物学特性变化

OBJECTIVE: To explore the changes in biological characteristics of adipose-derived stem cells (ASCs) in obese patients post successful weight loss, so as to provide a reference for the clinical application of these ASCs in refractory wound repair. METHODS: This study was an experimental study. Twelve obese patients (8 females and 4 males, aged (50±9) years) who underwent abdominal skin tightening surgery after successful weight loss and were admitted to the First Affiliated Hospital of Zhengzhou University from April 2021 to April 2023 were included in weight loss group, and 12 healthy volunteers (10 females and 2 males, aged (50±9) years) who underwent abdominal liposuction and facial fat grafting surgery during the same period in the same institution were included in healthy group. Adipose tissue was collected from patients in weight loss group and volunteers in healthy group, and ASCs were extracted. Experiments were conducted using ASCs at passages 4 and 5. Cell proliferation levels were assessed using the methyl thiazolyl tetrazolium assay at 0 (immediately), 24, 48, and 72 hours of culture. The cell scratch test was performed and the cell migration rates at 12 and 24 hours after scratching were calculated. The cell Transwell assay was performed and the number of migration cells at 24 hours after culture was counted. Adipogenic and osteogenic induction assays were carried out, and the adipogenic and osteogenic differentiation levels of cells were detected after 18 and 21 days of induction, respectively. Real-time fluorescence quantitative reverse transcription polymerase chain reaction was employed to measure the mRNA expressions of lipoprotein lipase (LPL), peroxisome proliferator-activated receptor gamma (PPARγ), Runt-related transcription factor 2 (Runx2), osteopontin, alkaline phosphatase (ALP), matrix metalloproteinase 9 (MMP-9), and transforming growth factor beta (TGF-β). The sample number of each experiment was 12. RESULTS: At 0 hour of culture, the cell proliferation levels of patients in weight loss group and volunteers in healthy group were 1.022±0.056 and 1.000±0.144, respectively, with no statistically significant difference between the groups (P > 0.05). At 24, 48, and 72 hours of culture, the cell proliferation levels of patients in weight loss group were 1.366±0.030, 1.353±0.012, and 1.390±0.016, respectively, which were significantly lower than 1.755±0.077, 1.737±0.014, and 1.700±0.023 of volunteers in healthy group (with t values of 16.27, 71.35, and 38.56, respectively, P values all < 0.05). In the cell scratch test, at 12 and 24 hours after scratching, the cell migration rates of patients in weight loss group were lower than those of volunteers in healthy group, but the differences were not statistically significant (P > 0.05). In the cell Transwell assay, after 24 hours of culture, there was no statistically significant difference in the number of migrated cells between patients in weight loss group and volunteers in healthy group (P > 0.05). After 18 days of adipogenic induction, the cell adipogenic differentiation level of patients in weight loss group was significantly lower than that of volunteers in healthy group (t=27.81, P < 0.05). After 21 days of osteogenic induction, the cell osteogenic differentiation level of patients in weight loss group was significantly lower than that of volunteers in healthy group (t=14.85, P < 0.05). Compared with those of volunteers in healthy group, the mRNA expressions of LPL, PPARγ, TGF-β, and Runx2 of patients in weight loss group were significantly reduced (with t values of 59.48, 146.10, 46.10, and 3.13, respectively, P < 0.05), while there were no statistically significant changes in the mRNA expressions of osteopontin, ALP, or MMP-9 (P > 0.05). CONCLUSIONS: Compared with healthy volunteers, the proliferative capacity of ASCs in obese patients after successful weight loss is significantly diminished, the differentiation potential is relatively weak, and the expression levels of some genes corresponding to adipogenic and osteogenic differentiation are decreased, which may affect the therapeutic efficacy of these ASCs in treating refractory wounds caused by burns, diabetes, or radiation injuries. Therefore, the donor differences of ASCs need to be considered in clinical application.