Cell–cell interaction in a coculture system consisting of CRISPR/Cas9 mediated GFP knock‐in HUVECs and MG‐63 cells in alginate‐GelMA based nanocomposites hydrogel as a 3D scaffold. Issue 8 (27th March 2020)

• Record Type: Journal Article
• Authors: Shahabipour, Fahimeh; Oskuee, Reza K.; Dehghani, Hesam; Shokrgozar, Mohammad A.; Aninwene, George E.; Bonakdar, Shahin
• Is Part Of: Journal of biomedical materials research. Volume 108:Issue 8(2020)
• Page Start: 1596
• Page End: 1606
• Publication Date: 2020-03-27
• Subjects: 3D coculture -- CRISPR/Cas9 -- GelMA‐alginate nanocomposite -- GFP‐knock in HUVECs; Biomedical materials -- Periodicals; 610.28
• DOI: 10.1002/jbm.a.36928
• Languages: English
• ISSNs: 1549-3296

Abstract

Abstract: The interaction between osteogenic and angiogenic cells through a coculturing system in biocompatible materials has been considered for successfully engineering vascularized bone tissue equivalents. In this study, we developed a hydrogel‐blended scaffold consisted of gelatin methacryloyl (GelMA) and alginate enriched with hydroxyapatite nanoparticles (HAP) to model an in vitro prevascularized bone construct. The hydrogel‐based scaffold revealed a higher mechanical stiffness than those of pure (GelMA), alginate, and (GelMA+ HAP) hydrogels. In the present study, we generated a green fluorescent protein (GFP) knock‐in umbilical vein endothelial cells (HUVECs) cell line using the CRISPR/Cas9 technology. The GFP was inserted into the human‐like ROSA locus of HUVECs genome. HUVECs expressing GFP were cocultured with OB‐like cells (MG‐63) within three‐dimensionally (3D) fabricated hydrogel to investigate the response of cocultured osteoblasts and endothelial cells in a 3D structure. Cell viability under the 3D cocultured gel was higher than the 3D monocultured. Compared to the 3D monocultured condition, the cells were aligned and developed into the vessel‐like structures. During 14 days of culture periods, the cells displayed actin protrusions by the formation of spike‐like filopodia in the 3D cocultured model. Angiogenic and osteogenic‐related genes such as CD31, vWF, and osteocalcin showed higher expression in the cocultured versus the monocultured. These results have collectively indicated that the 3D cocultured hydrogel facilitates interaction among cells, thereby having a greater effect on angiogenic and osteogenic properties in the absence of induction media.