Enhancing Fat Transplantation Efficiency in a Mouse Model through Pretreatment of Adipose-Derived Stem Cells with RIP3 Inhibitors.

BACKGROUND: Autologous fat transplantation, widely used in cosmetic and reparative surgery for volumetric enhancements, faces challenges with its inconsistent long-term survival rates. The technique's efficacy, crucial for its development, is hindered by unpredictable outcomes. Enriching fat grafts with adipose-derived stem cells (ADSCs) shows promise in improving survival efficiency. OBJECTIVES: This study aimed to explore the potential of receptor-interacting protein kinase 3 (RIP3) kinase inhibitors as a pretreatment for ADSCs in enhancing autologous fat graft retention over a long term. METHODS: ADSCs were isolated, cultured under normal or oxygen-glucose deprivation conditions, and mixed with particulate fat grafts to form distinct experimental groups in female nude mice. Fat graft mass and volume, along with underlying mechanisms, were evaluated using quantitative reverse transcription polymerase chain reaction (RT-qPCR), immunohistochemistry, and Western blot analysis. RESULTS: The experimental group, pretreated with RIP3 kinase inhibitors, had higher graft mass and volume, greater adipocyte integrity, and increased peroxisome proliferator-activated receptor gamma (PPARγ) mRNA levels than control groups. Furthermore, the experimental group demonstrated lower expression of necroptosis pathway proteins in the short term and an ameliorated inflammatory response as indicated by interleukin-1 beta (IL-1ß), interleukin-10 (IL-10) mRNA levels, and histological analyses. Notably, enhanced neovascularization was evident in the experimental group. CONCLUSIONS: These findings suggest that RIP3 kinase inhibitor pretreatment of ADSCs can improve fat graft survival, promote adipocyte integrity, potentially decrease inflammation, and enhance neovascularization. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each submission to which Evidence-Based Medicine rankings are applicable. This excludes Review Articles, Book Reviews, and manuscripts that concern Basic Science, Animal Studies, Cadaver Studies, and Experimental Studies. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

The Effect of Various Temperatures on the Inflammatory Profile of Fat Graft Storage: An Experimental Study.

Fat tissue has been widely used as a filler material during plastic surgery, but unpredictable fat retention remains a significant concern. Fat tissue is vulnerable to ischemia and hypoxia, but it always has waiting time before injection in the operation theater. Apart from transferring fat tissue as quickly as possible after harvesting, washing the aspirate with cool normal saline is often used. However, the mechanisms of cool temperature acting on adipose tissue have yet to be fully elucidated. Herein, this study aims to explore the effect of preservation at different temperatures on the inflammatory profile of adipose tissue. Inguinal adipose tissue of rats was collected and cultured in vitro under 4°C, 10°C, and room temperature for 2 hours. The proportion of damaged adipocytes and an array of cytokines were determined. We observed that the damage rate of the adipocyte membrane was slightly higher at room temperature, but there was no significant difference, while we noticed increased IL-6 and MCP-1 levels in adipose tissue at room temperature ( P ＜0.01). The 4°C and 10°C cool temperatures may offer protection against proinflammatory states during the adipose tissue preserved in vitro.

The histologic reaction and permanence of hyaluronic acid gel, calcium hydroxylapatite microspheres, and extracellular matrix bio gel.

BACKGROUND: The filling materials on the beauty market can be classified into three types: natural biological materials, synthetic polymer materials, and composites containing bioactive substances. However, comparative experimental data is lacking to compare their biological responses and permanence. AIMS: The main object of this study was to evaluate the biological response of these three types of fillers to provide a theoretical basis for clinical application. METHODS: Six-week-old female mice were injected subcutaneously with hyaluronic acid (HA) gel, calcium hydroxylapatite (CaHA) microspheres, and extracellular matrix (ECM) bio gel to observe the body reaction and permanence. At 1, 4, 8, and 16 weeks, the test sites were excised and analyzed by histopathology and proteomics. RESULTS: Extracellular matrix had a minimal foreign body response. HA had a good volume effect at the early stage but the volume retention rate was lower than CaHA in the long term. CaHA could stimulate neo-collagen formation. CONCLUSION: This study has proven the effectiveness and safety of these fillers and could provide clinical guidance for the plastic surgeon.

Breast Augmentation with Autologous Fat Grafting Immediately after Removal of Polyacrylamide Hydrogel and Fibrotic Capsule in 162 Patients.

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 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-Q™ 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. Level of Evidence: IV.

Biological characteristics of adipose-derived stem cells from patients with progressive hemifacial atrophy: An in vivo study.

OBJECTIVE: In this study, we investigate the biological characteristics of ADSCs from patients with progressive hemifacial atrophy (PHA) in vivo and try to explore the theoretical support for cell-assisted lipotransfer in treating patients with PHA. METHODS: ADSCs and exosomes were respectively extracted from patients with PHA and healthy ones. PHA-ADSC was detected the differentiation ability, phenotype, and anoxic resistance. NTA particle size, electron microscopy (TEM), and WB for CD63 and TSG10 were used to detect the exosomes. ADSCs of PHA (PHA-ADSCs) and healthy ones (NORM-ADSCs) mixed with their granular fat, exosomes mixed with PHA-granular fat, and PBS mixed with PHA-granular fat as the control group. The four groups of different grafts were, respectively, transplanted into nude mice, and the fat grafts were dissected and weighed at 2, 4, 8 and 12 w. Weight and volume retention were calculated for each of the four groups. Then, the four groups of fat grafts were tested for hematoxylin-eosin (HE) and immunohistochemical stainings, CD31 for blood vessel formation, CD68 for macrophage infiltration, and perilipin for fat formation, RT-PCR analysis of the APRC5, ATG5, ATG7, ATG12, BAX, PPARG, CDKN1A, and CDKN2A genes. RESULTS: ADSCs in the PHA group had typical phenotypes and multidirectional abilities. The PHA-ADSCs -assisted lipotransfer group, exhibited a weaker droplet formation and lower volume retention rate than the NORM-ADSCs-assisted lipotransfer group but much better than the non-cell-assisted lipotransfer group. Exosome-assisted lipotransfer group showed benefits, too. CONCLUSIONS: The PHA-ADSCs-assisted lipotransfer and the exosome-assisted lipotransfer improved the fat survival rate after fat filling in patients with hemifacial atrophy. Cell-assisted and exosome-assisted lipotransfer is an effective method to treat hemifacial atrophy.

In Vitro Study on the Biological Characteristics of Adipose-Derived Stem Cells from Liposuction Area in Patients with Progressive Hemifacial Atrophy.

OBJECTIVE: In this study, we investigate the biological characteristics of ADSCs from the liposuction area in patients with hemifacial atrophy in vitro. METHODS: ADSCs were respectively extracted from the donor site of patients with hemifacial atrophy and healthy ones. ADSCs of two groups were respectively tested for proliferation ability, phenotype, multipotency, migration ability, self-repair ability, apoptosis, and autophagy. Exosomes extracted from the supernatant of two groups were detected by NTA particle size, electron microscopy (TEM), and WB for CD63 and TSG10, respectively. RESULTS: CCK-8 showed a statistically less increase in cell proliferation in PHA-ADSCs after the sixth day. ADSCs in both groups had typical phenotypes and multidirectional abilities. PHA-ADSCs exhibited weaker droplet formation. The cell migration ability in PHA-ADSCs was weaker tested by Transwell assay. The live/dead proportion calculated by ImageJ following calcein-AM/PI double staining revealed live cells in PHA-ADSCs was 46.11% compared with 54.21% in NORM-ADSCs after OGD treatment. A significant down-regulation of ATG7 and ATG12 and a higher percentage of apoptosis were found in PHA-ADSCs. A significant up-regulation of BAX occurred in PHA-ADSCs.ARPC5 expression in the PHA group was extremely distinct down-regulated.CDKN1A and CDKN2A expression in the PHA group was significantly up-regulated.WB analyses confirmed that both groups' ADSCs-Exosomes surface markers CD63 and TSG101 were positively expressed but varied significantly. CONCLUSIONS: PHA-ADSCs exhibited a poorer proliferation ability, higher apoptosis percentage, weaker lipid droplets formation, weaker cell migration, poorer intolerance to OGD, aging earlier, and weaker self-renewal and repairability.PHA-ADSCs-Exosomes showed low expressions of CD63 and TSG101.This study provides strong evidence that the addition of exosomes with specific cytokines can improve the fat survival rate after fat filling in patients with hemifacial atrophy. NO LEVEL ASSIGNED: This journal requires that authors 42 assign a level of evidence to each submission to which 43 Evidence-Based Medicine rankings are applicable. This 44 excludes Review Articles, Book Reviews, and manuscripts 45 that concern Basic Science, Animal Studies, Cadaver 46 Studies, and Experimental Studies. For a full description of 47 these Evidence-Based Medicine ratings, please refer to the 48 Table of Contents or the online Instructions to Authors 49 https://www.springer.com/00266 .

Inhibition of RIP3 increased ADSC viability under OGD and modified the competency of adipogenesis, angiogenesis, and inflammation regulation.

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 (RIP3) and mixed lineage kinase domain-like pseudokinase (MLKL), were expressed in the early stage. The present 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 cell counting kit-8 (CCK-8), calcein/propidium iodide (PI) staining and immunostaining, we determined the OGD treatment time of 4 h, 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 γ (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, interleukin (IL)-10 (IL-10) and reduced the expression of the proinflammatory factor IL-1ß. 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.

Augmentation Mammoplasty With External Volume Expander-Assisted Autologous Fat Grafting in 305 Asian Patients.

BACKGROUND: External volume expander (EVE)-assisted autologous fat grafting is suitable for breast augmentation, but no large sample study in Asia has confirmed this method. OBJECTIVES: The authors reported their experience and outcomes in augmentation mammoplasty with EVE-assisted autologous fat grafting. METHODS: A retrospective study was conducted in 305 female patients who underwent augmentation mammoplasty with EVE-assisted fat grafting between September 2012 and December 2020. Doctors utilized Crisalix (Crisalix S.A., Lausanne, Switzerland) for 3-dimensional (3D) imaging acquisition to measure the increase in breast volume to evaluate doctor satisfaction. The Preoperative Satisfaction with Breast and BREAST-Q questionnaires were employed to assess patients' preoperative and postoperative satisfaction, respectively. RESULTS: The 305 female patients were aged 18 to 50 years (mean, 35.9 years). Among them, 68.52% were "very satisfied," 18.69% were "somewhat satisfied," 11.15% were "somewhat dissatisfied," and 1.64% were "very dissatisfied" based on BREAST-Q analysis, whereas 100% were dissatisfied according to the Preoperative Satisfaction with Breast questionnaire. Doctors employed Crisalix to measure the increase in breast volume to evaluate doctor satisfaction. The results showed 76.01% had an increase in breast volume of 150 to 250 mL or >250 mL and were "satisfied" and "very satisfied," respectively, 21.64% had an increase of 50 to 149 mL and were "somewhat satisfied," and 2.30% had an increase <50 mL and were "dissatisfied." There were no complications, such as obvious fat liquefaction, infection, or fat embolism. CONCLUSIONS: Augmentation mammoplasty with EVE-assisted fat grafting is effective and satisfying in China. Crisalix for 3D imaging acquisition is convenient and effective in measuring breast volume.