Abstract
Silk fibroin (SF), a natural polymer material possessing excellent biocompatibility and biodegradability, and has been widely used in biomedical applications. In order to explore the behavior of vascular cells by co-culturing on regenerated SF matrix for use as artificial blood vessels, human aorta vascular smooth muscle cells (HAVSMCs) were co-cultured with human arterial fibroblasts (HAFs) or human umbilical vein endothelial cells (HUVECs) on SF films and SF tubular scaffolds (SFTSs). Analysis of cell morphology and deoxyribonucleic acid (DNA) content showed that HUVECs, HAVSMCs and HAFs adhered and spread well, and exhibited high proliferative activity whether cultured alone or in co-culture. Immunofluorescence and scanning electron microscopy (SEM) analysis showed that HUVECs and HAFs co-existed well with HAVSMCs on SF films or SFTSs. Cytokine expression determined by reverse transcription-polymerase chain reaction (RT-PCR) indicated that the expression levels of α-smooth muscle actin (α-SMA) and smooth muscle myosin heavy chain (SM-MHC) in HAVSMCs were inhibited on SF films or SFTSs, but expression could be obviously promoted by co-culture with HUVECs or HAFs, especially that of SM-MHC. On SF films, the expression of vascular endothelial growth factor (VEGF) and platelet endothelial cell adhesion molecule-1 (CD31) in HUVECs was promoted, and the expression levels of both increased obviously when co-cultured with HAVSMCs, with the expression levels of VEGF increasing with increasing incubation time. The expression levels of VEGF and CD31 in cells co-cultured on SFTSs improved significantly from day 3 compared with the mono-culture group. These results were beneficial to the mechanism analysis on vascular cell colonization and vascular tissue repair after in vivo transplantation of SFTSs.
Highlights
Cardiovascular disease caused by thrombosis or a vascular lesion is a major cause of death of in humans
Cells of human aorta vascular smooth muscle cells (HAVSMCs), human umbilical vein endothelial cells (HUVECs) or co-cultures were all fully confluent on Silk fibroin (SF) films after 5 days, with HAVSMCs exhibiting a spindle shaped morphology (Figure 1A,B), and HUVECs spread in a typical pebble shape (Figure 1C,D) on film and tissue culture plate (TCP)
The results indicated that HAVSMCs and HUVECs could coexist on SF films without rejection or separation
Summary
Cardiovascular disease caused by thrombosis or a vascular lesion is a major cause of death of in humans. The development of blood vessel substitutes has rapidly progressed in the past decade in response to the clinical need. Vascular grafts have been realized to replace large-diameter (inner diameter > 6 mm) blood vessels, and these have been made from materials such as Dacron® [1]. Small-diameter (inner diameter < 6 mm) prosthetic vascular grafts for clinical applications remain unfeasible. Polyurethane [3], polyglycolic acid [4], polylactic acid [5], polycaprolactone [6] and their composites or derivatives [7,8] have been widely expected to be suitable for use as tissue-engineered vascular grafts prepared by Polymers 2018, 10, 39; doi:10.3390/polym10010039 www.mdpi.com/journal/polymers. Natural biopolymers are more conducive to cell adhesion, migration and proliferation and more suitable for biological material applications [13]
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