RESUMO
Platelet-rich plasma (PRP) has been widely used to improve the fat retention rate in autologous fat transplantation since it possesses a good angiogenesis capability in vivo. However, due to the short half-life of growth factors released from PRP and its uneven distribution in injected fat tissue, the strategy of PRP in fat transplantation needs further improvement. Since the capillaries started to grow into fat grafts in 1 week and vascular growth peaks in the second week after transplantation, we hypothesized that delayed two-steps PRP injection into the interior of grafts, accompanied with the extent of neovascularization might theoretically promote microvessel growth inside transplanted adipose tissue. 24 nude mice were divided into three groups: Blank group (0.35 mL fat mixed with 0.15 mL saline, N = 8), Single step group (0.35 mL fat mixed with 0.15 mLPRP, N = 8), and Two steps group (0.35 mL fat (day 0) + 0.075 mL PRP (day 7) + 0.075 mL PRP (day 14), N = 8). At 6 and 14 weeks post-transplantation, grafts were dissected, weighted, and assessed for histology, angiogenesis, fat regeneration and inflammation level. The weight and volume of the fat samples revealed no statistical difference among the three groups at 6 weeks after fat transplantation. The weight and volume of the Two steps group fat samples showed significantly higher compared to that in Blank and Single step groups at 14 weeks after fat transplantation (weight: 137.25 ± 5.60 mg versus 87.5 ± 3.90 mg,106.75 ± 2.94 mg, respectively; volume: 0.13 ± 0.01 mL versus 0.08 ± 0.01 mL, 0.09 ± 0.01 mL, respectively). Histological assessments indicated that delayed two-steps PRP injection strategy helps to improve adipose tissue content and reduce the composition of fibrous connective tissue at 14 weeks after fat transplantation. At 6 weeks and 14 weeks after transplantation, CD31 immunofluorescence indicated that delayed two-steps PRP injection strategy helps to improve angiogenesis and significantly higher compared to that in Blank and Single step groups (6 weeks: 28.75 ± 4.54 versus 10.50 ± 2.06, 21.75 ± 1.85; 14 weeks: 21.75 ± 2.86 versus 9.87 ± 2.08, 11.75 ± 1.47, respectively). Preadipocyte count indicated delayed two-steps PRP injection strategy might promote fat regeneration and significantly higher compared to that in Blank and Single step groups at 14 weeks (129.75 ± 6.57 versus 13.50 ± 3.50, 17.12 ± 6.23, respectively). In this study, we demonstrated that the novel delayed two-steps PRP injection strategy remarkably enhanced the long-term fat retention rate and improved the neovascularization extent in the interior of the fat graft. Platelet-rich plasma, Delayed two-steps injection, Angiogenesis, Fat transplantation.
Assuntos
Gordura Abdominal/irrigação sanguínea , Gordura Abdominal/transplante , Neovascularização Fisiológica/fisiologia , Plasma Rico em Plaquetas , Gordura Abdominal/citologia , Adipócitos/citologia , Adulto , Animais , Capilares , Sobrevivência Celular , Feminino , Meia-Vida , Humanos , Injeções , Macrófagos , Masculino , Camundongos Endogâmicos BALB C , Plasma Rico em Plaquetas/metabolismoRESUMO
OBJECTIVE: Adipose tissue (AT) dysfunction associated with obesity or aging is a major cause for lipid redistribution and the progression of cardiometabolic disorders. Our goal is to decipher the contribution of human AT microvascular endothelial cells (ECs) in the maintenance of fatty acid (FA) fluxes and the impact of senescence on their function. APPROACH AND RESULTS: We used freshly isolated primary microvascular ECs from human AT. Our data identified the endothelial FA handling machinery including FATPs (FA transport proteins) FATP1, FATP3, FATP4, and CD36 as well as FABP4 (FA binding protein 4). We showed that PPARγ (peroxisome proliferator-activated receptor gamma) regulates the expression of FATP1, CD36, and FABP4 and is a major regulator of FA uptake in human AT EC (hATEC). We provided evidence that endothelial PPARγ activity is modulated by senescence. Indeed, the positive regulation of FA transport by PPARγ agonist was abolished, whereas the emergence of an inflammatory response was favored in senescent hATEC. This was associated with the retention of nuclear FOXO1 (forkhead box protein O1), whereas nuclear PPARγ translocation was impaired. CONCLUSIONS: These data support the notion that PPARγ is a key regulator of primary hATEC function including FA handling and inflammatory response. However, the outcome of PPARγ activation is modulated by senescence, a phenomenon that may impact the ability of hATEC to properly respond to and handle lipid fluxes. Finally, our work highlights the role of hATEC in the regulation of FA fluxes and reveals that dysfunction of these cells with accelerated aging is likely to participate to AT dysfunction and the redistribution of lipids.
Assuntos
Gordura Abdominal/irrigação sanguínea , Senescência Celular , Células Endoteliais/metabolismo , Ácidos Graxos/metabolismo , Inflamação/metabolismo , Microvasos/metabolismo , PPAR gama/metabolismo , Transporte Ativo do Núcleo Celular , Proliferação de Células , Células Cultivadas , Inibidor p16 de Quinase Dependente de Ciclina/genética , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Células Endoteliais/ultraestrutura , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Ligação a Ácido Graxo/metabolismo , Feminino , Proteína Forkhead Box O1/metabolismo , Humanos , Inflamação/genética , Inflamação/patologia , Microvasos/ultraestrutura , PPAR gama/genética , Transdução de SinaisRESUMO
Fish oils (FOs) have anti-inflammatory effects and lower serum triglycerides. This study examined adipose and muscle inflammatory markers after treatment of humans with FOs and measured the effects of ω-3 fatty acids on adipocytes and macrophages in vitro. Insulin-resistant, nondiabetic subjects were treated with Omega-3-Acid Ethyl Esters (4 g/day) or placebo for 12 weeks. Plasma macrophage chemoattractant protein 1 (MCP-1) levels were reduced by FO, but the levels of other cytokines were unchanged. The adipose (but not muscle) of FO-treated subjects demonstrated a decrease in macrophages, a decrease in MCP-1, and an increase in capillaries, and subjects with the most macrophages demonstrated the greatest response to treatment. Adipose and muscle ω-3 fatty acid content increased after treatment; however, there was no change in insulin sensitivity or adiponectin. In vitro, M1-polarized macrophages expressed high levels of MCP-1. The addition of ω-3 fatty acids reduced MCP-1 expression with no effect on TNF-α. In addition, ω-3 fatty acids suppressed the upregulation of adipocyte MCP-1 that occurred when adipocytes were cocultured with macrophages. Thus, FO reduced adipose macrophages, increased capillaries, and reduced MCP-1 expression in insulin-resistant humans and in macrophages and adipocytes in vitro; however, there was no measureable effect on insulin sensitivity.
Assuntos
Gordura Abdominal/imunologia , Suplementos Nutricionais , Ácidos Graxos Ômega-3/uso terapêutico , Resistência à Insulina , Macrófagos/imunologia , Síndrome Metabólica/dietoterapia , Obesidade/complicações , Gordura Abdominal/irrigação sanguínea , Gordura Abdominal/metabolismo , Gordura Abdominal/patologia , Indutores da Angiogênese/metabolismo , Indutores da Angiogênese/uso terapêutico , Anti-Inflamatórios não Esteroides/metabolismo , Anti-Inflamatórios não Esteroides/uso terapêutico , Índice de Massa Corporal , Capilares/imunologia , Capilares/metabolismo , Capilares/patologia , Células Cultivadas , Quimiocina CCL2/sangue , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Técnicas de Cocultura , Ácidos Docosa-Hexaenoicos , Regulação para Baixo , Combinação de Medicamentos , Ácido Eicosapentaenoico , Ácidos Graxos Ômega-3/metabolismo , Feminino , Óleos de Peixe/uso terapêutico , Humanos , Macrófagos/metabolismo , Macrófagos/patologia , Masculino , Síndrome Metabólica/complicações , Síndrome Metabólica/imunologia , Síndrome Metabólica/metabolismo , Pessoa de Meia-Idade , Músculos/imunologia , Músculos/metabolismo , Músculos/patologia , RNA Mensageiro/metabolismoRESUMO
Recent research has shown that adipose tissues contain abundant MSCs (mesenchymal stem cells). The origin and location of the adipose stem cells, however, remain unknown, presenting an obstacle to the further purification and study of these cells. In the present study, we aimed at investigating the origins of adipose stem cells. α-SMA (α-smooth muscle actin) is one of the markers of pericytes. We harvested ASCs (adipose stromal cells) from α-SMA-GFP (green fluorescent protein) transgenic mice and sorted them into GFP-positive and GFP-negative cells by FACS. Multilineage differentiation tests were applied to examine the pluripotent ability of the α-SMA-GFP-positive and -negative cells. Immunofluorescent staining for α-SMA and PDGF-Rß (platelet-derived growth factor receptor ß) were applied to identify the α-SMA-GFP-positive cells. Then α-SMA-GFP-positive cells were loaded on a collagen-fibronectin gel with endothelial cells to test their vascularization ability both in vitro and in vivo. Results show that, in adipose tissue, all of the α-SMA-GFP-positive cells congregate around the blood vessels. Only the α-SMA-GFP-positive cells have multilineage differentiation ability, while the α-SMA-GFP-negative cells can only differentiate in an adipogenic direction. The α-SMA-GFP-positive cells maintained expression of α-SMA during multilineage differentiation. The α-SMA-GFP-positive cells can promote the vascularization of endothelial cells in three-dimensional culture both in vitro and in vivo. We conclude that the adipose stem cells originate from perivascular cells and congregate around blood vessels.
Assuntos
Gordura Abdominal/citologia , Linhagem da Célula , Células-Tronco/citologia , Gordura Abdominal/irrigação sanguínea , Actinas/genética , Adipogenia , Animais , Forma Celular , Células Cultivadas , Condrogênese , Feminino , Genes Reporter , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Humanos , Camundongos , Camundongos Nus , Camundongos Transgênicos , Neovascularização Fisiológica , Osteogênese , Regiões Promotoras Genéticas , Transplante de Células-TroncoRESUMO
Knowledge of abdominal anatomy is key to achieving optimal results in abdominoplasty. With adequate knowledge of the anatomy, the surgeon can tailor his or her techniques to fit the needs of the patient while still maximizing the blood supply to the abdominal flaps and minimizing complications.
Assuntos
Abdome/cirurgia , Parede Abdominal/anatomia & histologia , Procedimentos de Cirurgia Plástica/métodos , Gordura Abdominal/irrigação sanguínea , Parede Abdominal/irrigação sanguínea , Parede Abdominal/inervação , Contraindicações , Humanos , Tecido Linfoide/anatomia & histologiaRESUMO
BACKGROUND AND AIMS: Morphological changes in adipose tissue reflect functional disorders that correlate with cardiometabolic complications of obesity. The metabolic risks vary among the obese individuals. Furthermore, normal-weight individuals are not necessarily metabolically healthy. Therefore, the aim of this study was to analyze morphological characteristics of the abdominal adipose tissue in normal-weight and obese individuals in regards to metabolic risks. METHODS AND RESULTS: The study group consisted of 30 overweight or obese and 20 normal-weight women undergoing elective surgery. Women of each group were divided into metabolically healthy and metabolically obese, based on the homeostasis model assessment of insulin resistance (HOMA-IR), triglyceride, total-, LDL- and HDL-cholesterol levels. The size and numerical density of adipocytes, as well as volume density of blood vessels in subcutaneous and visceral adipose tissue were compared among subgroups. The results showed hypertrophy of adipocytes of visceral adipose tissue in metabolically obese normal-weight women. At the same time, metabolically healthy obese women had smaller adipocytes in both depots in comparison with "at risk" obese women. The lowest volume density of blood vessels correlated with the largest diameter of adipocytes in "at risk" obese women indicating hypoxic changes in visceral adipose tissue. The observed differences of the adipose tissue morphology did not correlate with considerable phenotypic differences within either the normal-weight or obese women group. CONCLUSION: Changes in adipocyte size, cellular and vascular density of adipose tissue in relation with metabolic disorders, regardless of nutritional level, suggest limited capacity of fat deposition and adipose tissue response to hypoxia.
Assuntos
Gordura Abdominal/patologia , Doenças Cardiovasculares/epidemiologia , Resistência à Insulina , Obesidade/patologia , Obesidade/fisiopatologia , Gordura Abdominal/irrigação sanguínea , Adipócitos/patologia , Adiposidade , Adulto , Contagem de Células , Tamanho Celular , Feminino , Humanos , Hipertrofia , Gordura Intra-Abdominal/irrigação sanguínea , Gordura Intra-Abdominal/patologia , Lipídeos/sangue , Pessoa de Meia-Idade , Obesidade/sangue , Sobrepeso/sangue , Sobrepeso/patologia , Sobrepeso/fisiopatologia , Fatores de Risco , Gordura Subcutânea Abdominal/irrigação sanguínea , Gordura Subcutânea Abdominal/patologiaRESUMO
BACKGROUND: The previously described "perfusion zones" of the abdominal wall vasculature are based on filling of the deep inferior epigastric artery (DIEA) and all its branches simultaneously. With the advent of the DIEA perforator flap, only a single or several perforators are included in supply to the flap. As such, a new model for abdominal wall perfusion has become necessary. The concept of a "perforator angiosome" is thus explored. METHODS: A clinical and cadaveric study of 155 abdominal walls was undertaken. This comprised the use of 10 whole, unembalmed cadaveric abdominal walls for angiographic studies, and 145 abdominal wall computed tomographic angiograms (CTAs) in patients undergoing preoperative imaging of the abdominal wall vasculature. The evaluation of the subcutaneous branching pattern and zone of perfusion of individual DIEA perforators was explored, particularly exploring differences between medial and lateral row perforators. RESULTS: Fundamental differences exist between medial row and lateral row perforators, with medial row perforators larger (1.3 mm vs. 1 mm) and more likely to ramify in the subcutaneous fat toward the contralateral hemiabdomen (98% of cases vs. 2% of cases). A model for the perfusion of the abdominal wall based on a single perforator is presented. CONCLUSION: The "perforator angiosome" is dependent on perforator location, and can mapped individually with the use of preoperative imaging.
Assuntos
Parede Abdominal/irrigação sanguínea , Artérias Epigástricas , Mamoplastia , Retalhos Cirúrgicos/irrigação sanguínea , Gordura Abdominal/irrigação sanguínea , Gordura Abdominal/diagnóstico por imagem , Músculos Abdominais/irrigação sanguínea , Músculos Abdominais/diagnóstico por imagem , Parede Abdominal/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Angiografia , Cadáver , Estudos de Coortes , Feminino , Humanos , Masculino , Microcirculação , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios XRESUMO
Adipokines play important regulatory roles in the pathophysiology of obesity and insulin resistance. We measured plasma and interstitial concentrations of the adipokines adiponectin, resistin, leptin, monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6) and interleukin-8 (IL-8) in subcutaneous, abdominal and femoral adipose tissue using calibrated, large-pore microdialysis technique in 8 healthy, lean men on 2 experimental days. The interstitial leptin concentration was 2.5-fold higher in subcutaneous, femoral than abdominal adipose tissue (P<0.05), but no regional differences were found for the remaining adipokines (P>0.05). Adiponectin and leptin concentrations were higher in plasma than subcutaneous adipose tissue (approximately 25-fold and approximately 2-fold, respectively, P<0.05), whereas MCP-1, IL-6 and IL-8 concentrations were higher in subcutaneous adipose tissue than plasma (approximately 100-fold, approximately 200-fold and approximately 1000-fold, respectively, P<0.05). Resistin concentrations did not differ significantly between compartments. Adipose tissue blood flow (ATBF) showed no regional difference (P>0.05). The intra- and inter-subject variations of all investigated adipokines as well as of ATBF were substantial (coefficient of variation: 4-177%). In conclusion, interstitial leptin concentrations are approximately 2.5-fold higher in subcutaneous, femoral than abdominal adipose tissue, which might be a potential mechanism behind the health-benefits of "pear-shape". Furthermore, subcutaneous adipose tissue has a marked production of pro-inflammatory adipokines.
Assuntos
Gordura Abdominal/metabolismo , Adipocinas/metabolismo , Fêmur/metabolismo , Gordura Subcutânea/metabolismo , Gordura Abdominal/irrigação sanguínea , Adiponectina/metabolismo , Adulto , Quimiocina CCL2/metabolismo , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Leptina/metabolismo , Masculino , Microdiálise , Resistina/metabolismo , Gordura Subcutânea/irrigação sanguínea , Adulto JovemRESUMO
BACKGROUND: Neuropeptide Y is a signaling molecule that was recently found to stimulate adipose tissue growth in vitro by means of a peripherally acting mechanism involving the neuropeptide Y2 receptor found on adipocytes and endothelial cells. This study aims to evaluate the translational applications of a neuropeptide Y2 receptor agonist for autologous fat grafting in plastic surgery. METHODS: Murine and primate animal models were used to investigate the proliferative effects of neuropeptide Y on adipose tissue. The effect of applying neuropeptide Y to subcutaneous tissues in mice and monkeys was assessed by magnetic resonance imaging, histology, and immunohistochemistry. The effect of neuropeptide Y on human fat xenograft survival and vascularity in athymic mice was measured by ultrasonography and immunohistochemistry. Six animals per group were used in murine experiments, and two animals were used in the pilot primate study. RESULTS: Neuropeptide Y stimulated growth of adipose tissues when applied subcutaneously in mice and monkeys, and increased human fat xenograft survival and vascularity in athymic mice at 3 months. CONCLUSIONS: These data provide in vivo evidence for a critical role for neuropeptide Y/neuropeptide Y2 receptor interactions in adipogenesis, and suggest neuropeptide Y2 receptor as a potential target for agonist compounds that can be used to enhance fat graft survival or stimulate de novo adipogenesis.
Assuntos
Gordura Abdominal/transplante , Adipogenia/fisiologia , Sobrevivência de Enxerto/fisiologia , Neuropeptídeo Y/metabolismo , Receptores de Neuropeptídeo Y/metabolismo , Gordura Abdominal/irrigação sanguínea , Gordura Abdominal/crescimento & desenvolvimento , Adipócitos/citologia , Adipócitos/fisiologia , Animais , Feminino , Humanos , Macaca mulatta , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Nus , Transplante/métodos , Transplante HeterólogoRESUMO
OBJECTIVE: Rapid weight loss with very-low-calorie diet (VLCD) is known to improve insulin sensitivity and decrease adipose tissue masses. The aim was to investigate the effects of VLCD on adipose tissue regional glucose uptake (rGU) and perfusion and their association with adipokines. RESEARCH DESIGN AND METHODS: Sixteen healthy obese (body mass index 33+/-1.1 kg/m(2)) subjects underwent VLCD for 6 weeks. RGU and perfusion were measured using [(18)F]-fluoro-deoxy-glucose, [(15)O]H(2)O and positron emission tomography. RESULTS: Blood-flow and rGU expressed per gram of adipose tissue were higher in visceral fat compared to abdominal subcutaneous fat (P<0.01 for both). Dieting decreased weight by 11+/-0.9 kg (P<0.0001). Visceral adipose fat decreased by 25% (P<0.001) and abdominal subcutaneous fat by 16% (P<0.001). Whole body insulin sensitivity increased by 33% (P<0.01). Perfusion of both fat depots decreased (P<0.001), while rGU remained unchanged. Among the adipokines, leptin and interleukin-6 levels seemed to be associated with abdominal subcutaneous and intra-abdominal adipose tissue insulin resistance but not with adipose tissue perfusion. CONCLUSIONS: Abdominal adipose tissue perfusion and rGU are not related in obesity. Rapid weight loss decreases perfusion through adipose tissue depots but has no influence on rGU demonstrating the 'sink' role of adipose tissue.
Assuntos
Gordura Abdominal/irrigação sanguínea , Gordura Abdominal/metabolismo , Glicemia/metabolismo , Resistência à Insulina/fisiologia , Gordura Intra-Abdominal/irrigação sanguínea , Gordura Intra-Abdominal/metabolismo , Obesidade/fisiopatologia , Redução de Peso/fisiologia , Adulto , Índice de Massa Corporal , Restrição Calórica , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Interleucina-6/sangue , Leptina/sangue , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Obesidade/metabolismo , Tomografia por Emissão de Pósitrons , Fluxo Sanguíneo RegionalRESUMO
INTRODUCTION: The peripheral adipose tissue is a vital component of many procedures in reconstructive and esthetic surgery. There is a limited knowledge on hemodynamic changes of adipose tissue. A direct intravital microscopic observation method is needed for the measurement of microcirculatory changes occurring in peripheral fat tissue during different plastic surgical procedures. Here we are introducing a model allowing for the direct in vivo monitoring and measurements of microcirculatory hemodynamics of peripheral adipose tissue. METHODS: Eight male Lewis rats weighing between 150 and 180 g were used in this study. Eight abdominal adipofascial flaps based on the left femoral artery, vein, and nerve were dissected after excision of the skin of the groin and lower abdominal region. Flap angiography using Indian ink was performed to demonstrate vascular anatomy of the flap. Standard intravital microscopy was used to monitor hemodynamic parameters such as vascular diameters, functional capillary perfusion, and leukocyte-endothelial interactions. RESULTS: Under direct intravital microscopy, microcirculatory parameters including vascular diameters, capillary perfusion, and leukocyte-endothelial interaction behaviors of the abdominal adipofascial flaps were established. CONCLUSIONS: We have showed feasibility of monitoring microcirculatory hemodynamics of the abdominal adipofascial flap model in rat. This model can be applied for intravital recordings of peripheral adipose tissue physiology and in different research scenarios such as the effects of ischemia reperfusion injury, effects of surgical trauma and wound healing studies with application of different pharmacologic agents and treatment protocols.