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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
Author(s) -
Shahabipour Fahimeh,
Oskuee Reza K.,
Dehghani Hesam,
Shokrgozar Mohammad A.,
Aninwene George E.,
Bonakdar Shahin
Publication year - 2020
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.36928
Subject(s) - self healing hydrogels , umbilical vein , cell culture , green fluorescent protein , microbiology and biotechnology , materials science , scaffold , tissue engineering , osteocalcin , in vitro , biophysics , chemistry , biology , biomedical engineering , biochemistry , medicine , alkaline phosphatase , gene , polymer chemistry , genetics , enzyme
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.