ABSTRACT
@#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.
ABSTRACT
<p><b>OBJECTIVES</b>To 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.</p><p><b>METHODS</b>A 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.</p><p><b>RESULTS</b>Light 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.</p><p><b>CONCLUSIONS</b>Porcine thoracic aortas ACTM can be obtained successfully through this procedure. Triton X-100 is a good reagent for preparing vessel ACTM.</p>