Adipose-derived stem cells (ADSCs) showed decreased cell viability and increased cell death under oxygen-glucose deprivation (OGD). Meanwhile, vital necroptotic proteins, including receptor-interacting protein kinase (RIP) 3 and mixed lineage kinase domain-like pseudokinase (MLKL), were expressed in the early stage. This study aims to explore the effect of necroptosis inhibition on ADSCs. ADSCs were obtained from normal human subcutaneous fat and verified by multidirectional differentiation and flow cytometry. By applying CCK8, calcein/PI staining and immunostaining, we determined the OGD treatment time of 4 hours, a timepoint when the cells showed a significant decrease in viability and increased protein expression of RIP3, phosphorylated RIP3 (pRIP3) and phosphorylated MLKL (pMLKL). After pretreatment with the inhibitor of RIP3, necroptotic protein expression decreased under OGD conditions, and cell necrosis decreased. Transwell assays proved that cell migration ability was retained. Furthermore, the expression of the adipogenic transcription factor peroxisome proliferator-activated receptor gamma (PPARγ) and quantitative analysis of oil red O staining increased in the inhibitor group. The expression of vascular endothelial growth factor-A (VEGFA) and fibroblast growth factor 2 (FGF2) and the migration test suggest that OGD increases the secretion of vascular factors, promotes the migration of human umbilical vein endothelial cells (HUVECs), and forms unstable neovascularization. ELISA revealed that inhibition of RIP3 increased the secretion of the anti-inflammatory factor IL-10 and reduced the expression of the proinflammatory factor IL-1β. Inhibition of RIP3 can reduce the death of ADSCs, retain their migration ability and adipogenic differentiation potential, reduce unstable neovascularization and inhibit the inflammatory response.
Objective In this study we investigated the feasibility and efficacy of immediate breast augmentation with autologous fat grafting after removal of polyacrylamide hydrogel(PAAG) and fibrotic capsule. Methods A retrospective study was conducted on 162 female patients who underwent removal of breast filler polyacrylamide hydrogel(PAAG) and the fibrotic capsule which produced after injection of PAAG via areola Omega-shaped incision. Then autologous fat grafting was immediately performed evenly and radially around the areola via the same incision into different layers (subcutaneous,submammary tissue,pectoralis major intramuscular and inferior pectoralis major space) except the empty cavity.The cavity left by removal of PAAG and fibrous capsule was closed with negative pressure drainage tube and slight external pressure. Results All patients recovered well without severe complications. The average score of postoperative satisfaction with physical well-being:chest was 99.83 (total score:100) compared with the average satisfaction score of 71.69 (total score:100) preoperatively by means of BREAST-QTM evaluation(P<0.01).All patients were satisfied with their postoperative breast shape. Conclusions Removing as much as possible is critical for patients who underwent the PAAG injection.Our experience in immediate breast augmentation with autologous fat grafting after removal of PAAG and fibrotic capsule proved useful and effective to maintain the balance between removing the PAAG as much as possible and retaining soft tissue to reshape breasts.
Background Platelet-rich fibrin (PRF) can promote fat graft survival. But the reported mixing ratio of PRF and fat ranges from 1:25 to 1:2, lacking a clear standard for clinical application. Objectives Explore the long-term effects of PRF on the grafted fat, and their optimal mixing ratio. Methods Nude mice were randomly divided into a control group (receiving subcutaneous injection of fat granules) and 4 PRF groups (receiving subcutaneous injection of PRF and fat granules at a volume ratio of 1:5, 1:10, 1:15, and 1:20, respectively). The graft samples (n=12) were obtained in weeks 4, 8 and 12 to (1) calculate retention rates, (2) evaluate gene and protein expression of VEGF-A, PPAR-γ, COL1-A1, and BAX, (3) perform H&E, Masson’s trichrome, α-SMA, and periplipin-1 stainings, and (4) count the microvessels and viable adipocytes. Results Compared with the control group, PRF groups had higher retention rates, a higher gene/protein expression of VEGF-A, a lower gene/protein expression of COL1-A1 and BAX, less fibrosis, and more microvessels and viable adipocytes. Group 1:10 was superior than other groups in terms of retention rates and other evaluation indexes. The expression of PPAR-γ had no significant difference among groups. Conclusions PRF may not play a long-term effect on adipogenesis. But it can still promote fat graft survival through facilitating vascularization, regulating collagen production and inhibiting apoptosis. PRF can achieve the best promoting effect when the mixing ratio of PRF and fat is 1:10, which is recommended as the optimal ratio for clinical application.
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