Comparison of immune responses of acellular vascular materials prepared by the two methods in animals

doi:10.3969/j.issn.2095-4344.2329

Abstract

Abstract:

BACKGROUND: As a mature method of decellularization at present, descaling agent has the advantages of simple operation, small damage, and easy control of experimental conditions. Some studies have combined different descaling agents to achieve good results in decellularization.

OBJECTIVE: To compare the effect of two descaling agents and the immune response of two kinds of acellular vascular materials subcutaneously implanted into animals.

METHODS: Two methods were used to treat the porcine carotid artery for descaling agent. One group was treated in the mixed solution of 1% sodium dodecyl sulfate and 1% sodium deoxycholate for 72 hours; the other group was treated in the mixed solution of 1% sodium dodecyl sulfate and 1% Triton X-100 for 72 hours. After cell removal, histological analysis, scanning electron microscope analysis, mechanical analysis and DNA content detection were carried out. Two groups of acellular vascular materials were implanted subcutaneously in the back of SD rats. After 1, 2, 4 and 8 weeks, the grafted vascular materials were taken out for hematoxylin and eosin and immunofluorescence staining. This study was approved by the Animal Ethics Committee of Jiangnan University.

RESULTS AND CONCLUSION: (1) Histological analysis showed that 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate effectively removed the cell components and retained the extracellular matrix structure, and the effect of cell removal was better than 1% sodium dodecyl sulfate combined with 1% Triton X-100. (2) The DNA content of 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate group was significantly lower than that of 1% sodium dodecyl sulfate combined with 1% Triton X-100 group (P < 0.05). There were no significant differences in bursting strength and suture tolerance between the two schemes (P > 0.05). (3) Scanning electron microscopy showed that 1% sodium dodecyl sulfate combined with 1% Triton X-100 significantly damaged the extracellular matrix compared with 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate. (4) In the subcutaneous implantation experiment, a large number of inflammatory cells infiltrated around the carotid artery materials of the two groups at 1 week after operation. No inflammatory cells infiltrated around the acellular materials of 1% sodium dodecyl sulfate and 1% sodium deoxycholate at 8 weeks after operation. Lymphocyte infiltrated around the acellular materials of 1% sodium dodecyl sulfate and 1% Triton X-100 group. 1% sodium dodecyl sulfate combined with 1% Triton X-100 acellular material mainly induced macrophages to differentiate into M1 type. 1% sodium dodecyl sulfate combined with 1% sodium deoxycholate acellular material mainly induced macrophages to differentiate into M2 type. (5) Compared with 1% sodium dodecyl sulfate and 1% Triton X-100, 1% sodium dodecyl sulfate and 1% sodium deoxycholate are more promising cell-free treatment.

Key words: material, experiment, protein, acellular blood vessel, descaling agent, immune response, pig carotid artery, macrophage

CLC Number:

Yin Jing, Wang Dan, Wang Peng, Zhang Hong, Jiang Haijun. Comparison of immune responses of acellular vascular materials prepared by the two methods in animals[J]. Chinese Journal of Tissue Engineering Research, 2020, 24(28): 4555-4561.

Figures/Tables 7

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