Aim:Unpredictable retention outcomes remain a significant issue in autologous fat grafting procedures.Liposuction cannula variation leads to variability in fat particle size.Recent data suggest that the size of fat pa...Aim:Unpredictable retention outcomes remain a significant issue in autologous fat grafting procedures.Liposuction cannula variation leads to variability in fat particle size.Recent data suggest that the size of fat particles is closely related to graft healing outcomes;however,this remains a point of contention due to potential confounding variables such as tissue trauma with harvest.The aim of this study was to compare autologous fat grafting outcomes with variable fat particle sizes in an animal model which isolated fat particle size as the primary experimental variable.The overall goal of this work was to determine if reducing fat particle size is an effective method for enhancing graft retention in autologous fat grafting.Methods:The range of fat particle diameter harvested by four common liposuction cannulas was quantified to define relevant small and large particle target diameters.To determine if particle size impacted nutrient and oxygen permeability,small and large particles were incubated in vitro in a spinner flask with an abundance of culture media and vascular endothelial growth factor secretion was measured with enzyme-linked immunosorbent assay.Finally,small and large fat grafts were prepared from subcutaneous mouse fat pads and grafted in syngeneic Balb/CJ mice.Weight and volume retention were evaluated at 1,4,8,and 12 weeks.Histological analysis with Masson's trichrome and perilipin immunofluorescent staining was performed.Real-time quantitative polymerase chain reaction was performed foradipogenic,inflammatory and apoptotic genes.Results:The range of fat particle diameters harvested with four commonly used cannulas was 2-7 mm.In vitro studies showed that 5-7-mm particles had significantly increased VEGF secretion normalized to weight,indicating increased tissue hypoxia in these particles compared to 2-4-mm particles.Surprisingly,in vivo comparison in two unique studies showed 2-4-mm and 5-7-mm fat particles had comparable graft retention(P=0.5329).Masson's trichrome staining revealed increased extracellular matrix and fibrosis in the 5-7-mm particle group(P=0.0115).Adipocyte survival with perilipin demonstrated comparable viability.Gene expression showed large particles experienced increased inflammation and apoptosis at one week after grafting,but overall there were no significant differences between groups.Conclusion:The ideal fat particle size should be large enough to contain adequate mesenchyme while not so thick as to preclude imbibition.This study suggests that,despite changes in hypoxia and VEGF levels,differing fat particles(2-4-mm and 5-7-mm)can achieve similar graft retention.展开更多
基金This work was funded by a Pilot Funding Program for Early Stage Medical Technology Research and Development grant from the Center for Medical Innovation,Swanson School of Engineering,University of Pittsburgh(award no.F_062-2013)by National Institutes of Health for Dr.Rubin,Department of Plastic SurgeryUniversity of Pittsburgh School of Medicine(award no.5R01CA114246).
文摘Aim:Unpredictable retention outcomes remain a significant issue in autologous fat grafting procedures.Liposuction cannula variation leads to variability in fat particle size.Recent data suggest that the size of fat particles is closely related to graft healing outcomes;however,this remains a point of contention due to potential confounding variables such as tissue trauma with harvest.The aim of this study was to compare autologous fat grafting outcomes with variable fat particle sizes in an animal model which isolated fat particle size as the primary experimental variable.The overall goal of this work was to determine if reducing fat particle size is an effective method for enhancing graft retention in autologous fat grafting.Methods:The range of fat particle diameter harvested by four common liposuction cannulas was quantified to define relevant small and large particle target diameters.To determine if particle size impacted nutrient and oxygen permeability,small and large particles were incubated in vitro in a spinner flask with an abundance of culture media and vascular endothelial growth factor secretion was measured with enzyme-linked immunosorbent assay.Finally,small and large fat grafts were prepared from subcutaneous mouse fat pads and grafted in syngeneic Balb/CJ mice.Weight and volume retention were evaluated at 1,4,8,and 12 weeks.Histological analysis with Masson's trichrome and perilipin immunofluorescent staining was performed.Real-time quantitative polymerase chain reaction was performed foradipogenic,inflammatory and apoptotic genes.Results:The range of fat particle diameters harvested with four commonly used cannulas was 2-7 mm.In vitro studies showed that 5-7-mm particles had significantly increased VEGF secretion normalized to weight,indicating increased tissue hypoxia in these particles compared to 2-4-mm particles.Surprisingly,in vivo comparison in two unique studies showed 2-4-mm and 5-7-mm fat particles had comparable graft retention(P=0.5329).Masson's trichrome staining revealed increased extracellular matrix and fibrosis in the 5-7-mm particle group(P=0.0115).Adipocyte survival with perilipin demonstrated comparable viability.Gene expression showed large particles experienced increased inflammation and apoptosis at one week after grafting,but overall there were no significant differences between groups.Conclusion:The ideal fat particle size should be large enough to contain adequate mesenchyme while not so thick as to preclude imbibition.This study suggests that,despite changes in hypoxia and VEGF levels,differing fat particles(2-4-mm and 5-7-mm)can achieve similar graft retention.
