Objective To prepare the acellular carotid artery of adult pig using biotic enzymes for small-diameter vascular tissue-engineering scaffold.Methods Carotid artery of adult pig was removed cells by using Trypsin/EDTA,ribonuclease and desoxyribonuclease.Residual cellular composition was studied with quantitative DNA analysis and the Hematoxylin and Eosin(H&E)staining.Extracellular matrix composition was evaluated with elastin van Gieson's(VG)staining and scanning electron microscopy(SEM).Results Quantitative DNA analysis and H&E staining confirmed that carotid arteries were completely decellularized.Elastin van Gieson's staining demonstrated that elastin morphology appeared unchanged.Scanning electron microscopy examination of the acellular scaffolds revealed a well-oriented porous decellularized structure that maintained natural architecture of the aorta.Conclusion Carotid artery of adult pig rendered acellular with Trypsin/EDTA,ribonuclease and desoxyribonuclease has well-preserved extracellular matrix for vascular scaffold,which can be used as a scaffold for further small-diameter vascular tissue engineering.

OBJECTIVESTo investigate the method of preparing porcine thoracic aortas acellular tissue matrix (ACTM) by trypsin, EDTA and Triton X-100 and to find the best concentration of X-100.

METHODSA total of 56 roots of fresh thoracic aortas (without adventitial tissue) from 80 kg-100 kg tame pigs were divided randomly into > groups, each containing 8 roots. Every vessel was put into a 50 ml centrifugal tube with a solution of 0.1% trypsin + 0.02EDTA in PBS for 24 h. After that, each group was separately immerged into a solution of 0.1%, 0.2%, 0.5%, 1.0%, 2.0%, 5.0%, 10.0% Triton X-100 for 144 h-240 h. Specimens were taken every 6 h. Specimens were stained with haematoxylin-eosin and observed grossly under the light and transmission electron microscopy.

RESULTSLight and transmission electron microscopy revealed that ACTM was composed of insoluble collagen, elastin, and some insoluble metamorphic organelles. The best concentration of Triton X-100 was 1% at the time of 176.25 h +/- 5.5 h.

CONCLUSIONSPorcine thoracic aortas ACTM can be obtained successfully through this procedure. Triton X-100 is a good reagent for preparing vessel ACTM.

Animals ; Aorta, Thoracic ; cytology ; surgery ; ultrastructure ; Blood Vessel Prosthesis ; Octoxynol ; pharmacology ; Swine ; Tissue Engineering ; methods