Evaluation of Vascular Endothelial Growth Factor Gene Expression in Recellularized Liver Tissue by Mouse Embryo Fibroblast

Authors
Motahare Homayoon Vala 1
Hamed Bagheri 2
Zinat Sargazi 3
Negar Bakhtiary 4
Shahram Pourbeiranvand 5
Mojdeh Salehnia 1
1 Department of Biomaterials, Faculty of Interdisciplinary Science and Technology, Tarbiat Modares University, Tehran, Iran
2 Department of Biomaterials, Faculty of Interdisciplinary Science and Technology, Tarbiat Mdares University, Tehran, Iran
3 Department of Anatomical Sciences, School of Medical Sciences, Zahedan University, Zahedan, Iran
4 1Department of Biomaterials, Faculty of Interdisciplinary Science and Technology, Tarbiat Modares University, Tehran, Iran
5 Department of Anatomical Sciences, School of Medical Sciences, Tarbiat Modares University, Tehran, Iran
10.61186/ibj.3862
Abstract
Background: The aim of the present study was to evaluate alterations in the vegf gene expression as an angiogenic factor in mouse embryo fibroblasts seeded on the decellularized liver fragments.

Methods: Liver tissue samples (n = 10) collected from adult male mice were randomly divided into decellularized and native control groups. Tissues were decellularized by treating with 1% Triton X-100 and 0.1% SDS for 24 hours and assessed by H&E staining and scanning electron microscopy (SEM). Then DNA content analysis and toxicity tests were performed. By centrifugation, DiI-labeled mouse embryo fibroblasts were seeded on each scaffold and cultured for one week. The recellularized scaffolds were studied by H&E staining, SEM, and laser scanning confocal microscopy (LSCM). After RNA extraction and cDNA synthesis, the expression of the vegf gene in these samples was investigated using real-time RT-PCR.

Results: Our observations showed that the decellularized tissues had morphology and porous structure similar to the control group, and their DNA content significantly reduced (p < 0.05) and reached to 4.12% of the control group. The MTT test indicated no significant cellular toxicity for the decellularized scaffolds. Light microscopy, SEM, and LSCM observations confirmed the attachment and penetration of embryonic fibroblast cells on the surface and into different depths of the scaffolds. There was no statistically significant difference in terms of vegf gene expression in the cultured cells in the presence and absence of a scaffold.

Conclusion: The reconstructed scaffold had no effect on vegf gene expression. Decellularized mouse liver tissue recellularized by embryonic fibroblasts could have an application in regenerative medicine.
Keywords
vascular endothelial growth factor
Gene expression
Fibroblast
Liver
Decellularized ECM