Behavior of Encapsulated MG-63 Cells in RGD and Gelatine-Modified Alginate Hydrogels

Abstract

Achieving cell spreading and proliferation inside hydrogels that are compatible with microencapsulation technology represents a major challenge for tissue engineering scaffolding and for the development of three-dimensional cell culture models. In this study, microcapsules of 650-900 μm in diameter were fabricated from oxidized alginate covalently cross-linked with gelatine (AlGel). Schiff's base bond formed in AlGel, detected by Fourier transform infrared spectroscopy, which confirmed the cross-linking of oxidized alginate with gelatine. Biological properties of alginate based hydrogels were studied by comparing the viability and morphology of MG-63 osteosarcoma cells encapsulated in gelatine and RGD-modified alginate. We hypothesized that the presence of gelatine and RGD will support cell adhesion and spreading inside the microcapsules and finally, also vascular endothelial growth factor (VEGF) secretion. After 4 days of incubation, cells formed extensive cortical protrusions and after 2 weeks they proliferated, migrated, and formed cellular networks through the AlGel material. In contrast, cells encapsulated in pure alginate and in RGD-modified alginate formed spherical aggregates with limited cell mobility and VEGF secretion. Metabolic activity was doubled after 5 days of incubation, making AlGel a promising material for cell encapsulation.

3D bioprinting of liver-mimetic construct with alginate/cellulose nanocrystal hybrid bioink

“…There is an ever-growing amount of evidence reported in research literature that gelatin is effective in promoting the cell attachment due to its RGD tripeptide sequences, which are the major integrin-binding domains present within extracellular matrix (ECM). 33,34 Our preliminary results ( Figure S4) show that the incorporation of gelatin in the alginate-CNC hybrid bioink does not influence the viscosity and hence printability of the bioink. Figures 5D-E show the fluorescence images of homogeneously distributed fibroblasts and human hepatoma cells in the bioprinted constructs.…”

Biofabrication of 3D Alginate-Based Hydrogel for Cancer Research: Comparison of Cell Spreading, Viability, and Adhesion Characteristics of Colorectal HCT116 Tumor Cells

“…Our 3D system can mimic the structural architecture and biological functions of the ECM in regard to mechanical properties and chemical composition and supports cell growth and survival (10).…”

“…Thus, the ADA-GEL matrix possibly provided more space for the cells to proliferate. Moreover, it composes a lower network density than the pristine alginate, indicated by the lower mechanical properties as reported(10). This makes it a preferable matrix for creating 3D tumor models.…”

“…Cells modify their metabolism and functionality in 2D cell culture systems since contacts with other cells and with their microenvironments cannot be formed (7,8). To mimic the structural architecture and cellular functions of the extracellular matrix (ECM) in 3D model systems some major characteristics have to be considered: suitable mechanical characteristics and chemical composition, facilitation of cell growth and maintenance, and fostering the transport of nutrients, gas, and metabolic waste, as well as signal transduction (7,9,10). The main advantage of 3D cultures in comparison to 2D monolayers is their clearly defined geometry that can help to co-culture different cell types with diverse phenotypes that are often found in solid epithelial tumors (2).…”

Targeted osteosarcoma chemotherapy using RGD peptide-installed doxorubicin-loaded biodegradable polymeric micelle

“…The affinity of RGD towards osteosarcoma cells brings inspiration to Sun et al [21] and Grigore et al [22] incorporated RGD motifs into hydrogels to generate cellular adhesion sites to enhance osteocytes (MG-63 was used as model cell line in their reports) spreading and differentiation for guiding tissue formation, as well as Haubner et al used 18 F-labeled RGD-containing glycopeptide for murine osteosarcomas imaging [23]. However, to the best of our knowledge, few RGD-associated nanosystems were explored for targeted chemotherapy of osteosarcoma.…”