Acquistion of a suitable extracellular matrix scaffold for tissue-engineered small-diameter blood vessel using a novel decellularization protocol

doi:10.3969/j.issn.2095-4344.1570

Abstract

Abstract:

BACKGROUND: A proper scaffold is the basis for construction of tissue-engineered small diameter blood vessels. Preliminary study has shown non-ionic detergents with limited effects of decellularization, and combined application of ionic detergents exhibits more effective decellularization.

OBJECTIVE: To compare the effectiveness of different preferable methods for preparing porcine carotid artery derived extracellular matrix scaffold, and to explore the optimal preparation method.

METHODS: Porcine carotid arteries (n=30) were obtained and randomized into five groups (n=6 per group). Then porcine carotid arteries were decellularized, followed by treatment with 1% sodium dodecyl sulfate and 1% sodium deoxycholate (group B), 0.5% sodium dodecyl sulfate and 0.5% sodium deoxycholate (group C), 1% sodium dodecyl sulfate and 1% Triton X-100 (group D), 0.5% sodium dodecyl sulfate and 0.5% Triton X-100 (group E). Native procine carotid aitery served as control (group A). Tissue samples of each group underwent hematoxylin-eosin staining, and Movat’s pentachrome staining, scanning electron microscope, immunohistochemistry and biomechanical test.

RESULTS AND CONCLUSION: Hematoxylin-eosin staining and Movat’s staining revealed that the cellular components were completely removed in the groups B and C; however, cellular residues were visualized in the groups D and E. Under the scanning electron microscope collagen and elastin fiber bundles were well preserved in the decellularization groups; however, the morphology and size of porous structure in the group B were superior to those in the groups C, D and E. Immunohistochemistry staining showed that the xenogeneic antigen, α-1,3-Gal, expressed in the decellularization groups; however, the expression of α-1,3-Gal was lower in the group B than the other groups. Biomechanical test indicated that burst pressure, suture-holding capacity and suture-holding strength were significantly decreased in the decellularization groups compared with group A (P < 0.05). However, there was no significant difference in the compliance, longitudinal elongation at brake, circumferential ultimate tensile stress, circumferential elongation at break and circumferential Young’s modulus compared with control group (P > 0.05). Decellularization caused a significant decrease in the longitudinal ultimate tensile stress and longitudinal Young’s modulus in the decellularization groups (P < 0.05). Furthermore, the longitudinal ultimate tensile stress and longitudinal Young’s modulus in the groups B and C were significantly lower than those in the groups D and E (P < 0.05). Our results suggest that the novel decellularization method based on 1% sodium dodecyl sulfate and 1% sodium deoxycholate optimizes the extracellular matrix scaffold for the construction of tissue-engineered small-diameter blood vessel.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Extracellular Matrix;, Blood Vessel Prosthesis, Biomechanics, Tissue Engineering

CLC Number:

Pu Lei, Pan Xingna, Zhang Jing, Wu Jian, Li Yaxiong, Yang Yingnan. Acquistion of a suitable extracellular matrix scaffold for tissue-engineered small-diameter blood vessel using a novel decellularization protocol[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(6): 855-862.

Figures/Tables 8

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