Thermal inkjet bioprinting triggers the activation of the VEGF pathway in human microvascular endothelial cells in vitro

Abstract

One biofabrication process that has gained tremendous momentum in the field of tissue engineering and regenerative medicine is cell-printing or most commonly bioprinting. We have shown that thermal inkjet bioprinted human microvascular endothelial cells were recruited or otherwise involved in the formation of microvasculature to form graft-host anastomoses upon implantation. The present study aims to quantify and characterize the expression and activation of specific cytokines and kinases in vitro. Morphological characteristics demonstrate elongated protrusions of TIB-HMVECs at 5-6 times the size of manually pipetted cells. Moreover, annexin V-FITC and propidium iodide apoptosis assay via flow cytometry demonstrated a 75% apoptosis among printed cells as compared to among control cells. Cell viability at a 3 d incubation period was significantly higher for printed cells as compared to control. Milliplex magnetic bead panels confirmed significant overexpression of HSP70, IL-1α, VEGF-A, IL-8, and FGF-1 of printed cells compared to control. In addition, a Human phospho-kinase array displayed a significant over activation of the heat-shock proteins HSP27 and HSP60 of printed cells compared to the manually seeded cells. Collectively, it is suggested that the massive appearance of capillary blood vessels upon implantation that has been reported elsewhere may be due to the activation of the HSP-NF-κB pathway to produce VEGF. This cell activation may be used as a new strategy for vascularization of tissue engineered constructs which are in high demand in regenerative medicine applications.

Thermal Bioprinting Causes Ample Alterations of Expression of LUCAT1, IL6, CCL26, and NRN1L Genes and Massive Phosphorylation of Critical Oncogenic Drug Resistance Pathways in Breast Cancer Cells

“…Increased levels of heat shock proteins may trigger a cell survival/resistance mechanism in cancer cells. Additionally, HSPs activate the angiogenic pathway that leads to vascular endothelial growth factor (VGEF) expression, vessel formation, and accelerated tumor formation in host tissues (Chatterjee et al, 2018;Plimpton et al, 2018;Solis et al, 2019). Jun N-terminal Kinases (JNKs) are small proteins, 45-55 kDa, and product of three genes which, through different splicing, create up to 10 isoforms (Alvarez et al, 1991;Hibi et al, 1993).…”

3D Bioprinting for Vascularized Tissue-Engineered Bone Fabrication

“…Inkjet bioprinting is a rapid and large-scale fabrication technique that has been adapted from the inkjet printing technology to print living cells by desktop inkjet printers [76] (Figure 3a). Currently, many studies utilize inkjet bioprinting to construct various engineered tissues with different kinds of living cells [77][78][79]. In addition, inkjet bioprinting can rapidly fabricate tissue-like structures with intricate hierarchical architectures by the utilization of the controlled dropwise deposition of cell-laden bioinks [79].…”

Emulating Human Tissues and Organs: A Bioprinting Perspective Toward Personalized Medicine

“… 100 Consequently, thermal jetting is a popular tool for cell-based bioprinting, particularly in applications requiring high levels of control over the spatial orientation of cells such as osteochondral, vascular, and oncological models. 101 − 103 …”