Immunogenicity and intensity of homegraft decelluladzed scaffolds without pretreatment
- VernacularTitle:未经预处理同种活性生物脱细胞支架的免疫原性及支架张力
- Author:
Wei WANG
;
Lilin MA
;
Liping LU
;
Ruixin CHEN
;
Xiu YU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2008;12(41):8193-8197
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:Pretreatment is a dominant way to increase the histocompatibility of biomaterials,but many pretreatments are not satisfactory due to some reactions,such as calcification,cytotoxic reaction and reducing the tension resistance.OBJECTIVE:To investigate the effect of immunogenicity,scaffold tension and cell growth factor on the non-pretreated scaffold biomaterials,and to prepare the homograft decellularized scaffolds.DESIGN:Contrast observation for the histomorphology.SETTING:The experiments were carried out in the Department of General Surgery,Affiliated Hospital of Nantong University from June 2006 to March 2007.MATERIALS:Wistar rats of SPF grade were adopted in this study.Sodium dodecyl sulfate from BioteehGrade Company(U.S.A.);basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) from Peprotech Company(UK);dynamometer from Suzhou Electrical Element First Factory (China).METHODS:The vena cava inferior from rats were selected as experimental materials,we decellularized the epithelial cells from vein according to the improved Booth's methods with low osmotic Tris buffer containing sodium dodecyl sulplate,and then reserved the extracellular matrix.After fixing,the decellularized scaffold was stained by hematoxylin and eosin,the collagen fibers were observed and photographed by light microscope and scanning electron microscope.Changes of the intensity of the scaffolds were measured before and after decellularization.The deceilularized scaffolds were transplanted subcutaneously at homogeneity rats to observe whether the scaffolds had induced immunological rejection.In combination with bFGF and/or VEGF,the scaffolds were transplanted subcutaneously at homogeneity rats and took out two weeks later,in order to exam the new vessels in the scaffolds by immunohistochemistry.Dynamometer was applied to compare the change of the intensity of the scaffolds before and after transplantation.MAIN OUTCOME MEASURES:Change of the intensity of the scaffolds after decellularization and transplantation.Formation of new vessels in scaffolds.Local rejection of the scaffold subcutaneously transplanted into rats.RESULTS:The endothelial cells of the vein were completely removed when incubated with Tris buffer containing 0.03% sodium dodecyl sulfate for 48 hours.Results of hematoxylin and eosin staining,light microscope and scanning electron microscope showed that,the main components of extracellular matrix (collagen fiber) were reserved.There was no significant difference in the morphological structure and the intensity of collagenous scaffolds before and after decellalarization (P＞0.05).There was no apparent reject reaction at the embedded site,local incisions healed well.New vessels could be observed in the scaffolds two weeks later.There was no significant difference in the intensity of collagenons scaffolds before and after inoculation (P＞0.05).CONCLUSION:The low osmotic Tris buffer containing 0.03% sodium dodecyl sulfate is suitable for decellularizing the vein.The homograft collagenous scaffolds exhibit no rejection after transplantation,and the intensity of the scaffolds does not descend.VEGF and bFGF can improve the growth of new vessels in the decellularized scaffolds,and have a synergistic effect.
