Comparison Of Adipogenesis Between Subcutaneous And Visceral Adipose Acellular Matrix Of Pigs And Their Mechanism

BackgroundSoft tissue defect is always the difficult problem in plastic surgery.Traditional therapeutic measures have many shortcomings and the application of biological materials has disadvantages such as absorption,low biocompatibility and rejection.Decellularization of adipose tissues(DAT),which are dealed with the decellular technology,can still retain the three-dimensional extracellular matrix structure.In the animal experiment,we found that they could be induced into adipose tissues.DAT are currently the important source of scaffolds in dipose tissue engineering.Porcine adipose tissue is widely available and easy to obtain.It is an important source of materials for studying DAT.Pig adipose tissue are divided into subcutaneous adipose tissue(SAT)and visceral adipose tissue(VAT),which are the best material source?Studies have pointed out that there are some physical structure differences between subcutaneous adipose tissue and visceral adipose tissue.We found in the preliminary experiment that SAT and VAT have different rigidity.Studies have pointed out that the mechanical environment of the extracellular matrix plays an important role in the growth of the cells,the rigidity of extracellular matrix significantly affect the behavior of the cells.Whether the rigidity of DAT affect their adipogenic capability in the regulation of adipos regeneration process?If we can prove that the two different rigiditie of the DAT scaffolds can induce different adipose tissue regeneration results,then how does the rigidity of the scaffolds affect the process of adipose tissue regeneration?This study based on the two kinds of DAT scaffolds as important experiment model,to explore the effect of the rigidity of decellularization of adipose tissues on adipogenic capability,aimed at affecting adipogenic capability of DAT scaffolds by adjusting the rigidity of them,So as to develop injectable filler of DAT.Methods and resultsWe used subcutaneous adipose tissue and visceral adipose tissue as source materials,and used physical homogenate method to deal with adipose tissues in advance to destroy the cell membranes,so as to release the oil fastly,saving time and reagent.The rigidity of DAT scaffolds testing result showed that the rigidity of the subcutaneous DAT was higher than that of the visceral DAT.These DAT scaffolds prepared by the above methods were placed in mice,and the materials were taken from 6 weeks and 12 weeks after transplantation.HE and immunofluorescence histologic results found that 6 weeks,12 weeks after transplantation,the adipocyte number of the visceral DAT was more than that of the subcutaneous DAT,Real,Time PCP detection results showed that 6 weeks,12 weeks after grafting,PPARyexpression of the visceral DAT is obviously higher than that of subcutaneous DAT graft.It prompt that two different rigidity of DAT scaffolds induce different adipose regeneration processes in vivo,and the visceral DAT scaffold have better adipogenic capability.Further in vitro experiment,we planted ASCs on subcutaneous DAT scaffolds and visceral DAT scaffolds.The cytoskeleton staining results showed that the cytoskeleton F-Actin of ASCs which on the subcutaneous DAT scaffold and visceral DAT scaffolds were all clustered in 3 days after planting.The cytoskeleton F-Actin of ASCs which on the subcutaneous DAT scaffold arranged more tidy but The cytoskeleton F-Actin of ASCs on visceral DAT scaffolds showed diffuse distribution in 14 days after planting.It meaned that the rigidity of DAT scaffolds effected adipogenic capability of ASCs by re-arrangement the cytoskeleton F-Actin of ASCs.Conclusions1.The physical homogenization method is an efficient method for the removal of oil from adipose tissue.2.The rigidity of SDAT scaffold is higher than that of VDAT scaffold.3.The adipogenic capability of VDAT scaffold is higher than that of SDAT scaffold.4.The rigidity of DAT scaffolds effected adipogenic capability of ASCs by re-arrangement the cytoskeleton F-Actin of ASCs.