Gellan Gum Hydrogels containing Extracellular Matrix (dECM) from Human Dermal Fibroblasts

last updated: 2021-12-16
ProjectECM_INK :: publications list
TitleGellan Gum Hydrogels containing Extracellular Matrix (dECM) from Human Dermal Fibroblasts
Publication TypeComunications - Poster
Year of Publication2019
AuthorsReis D. P., Fidalgo C., Gasperini L., Reis R. L., and Marques A. P.

Gellan Gum (GG), although commonly used in tissue engineering approaches due to its good biocompatibility and stability under physiological conditions, is biologically inert. To address this matter, the aim of this work was to investigate the influence of the incorporation of extracellular matrix (ECM) components over the cellular response to GG hydrogels. Human dermal fibroblasts (hDFBs) were cultured for 21 days in culture medium supplemented with 50 µg/ml ascorbic acid. An already established protocol based in freezing/thawing and DNase incubation was carried out to extract the maximum amount of ECM produced. Protein extracts were combined with GG to form hydrogels that were further crosslinked with PBS or culture medium. An array of formulations of GG/ECM with several polymer and ECM ratios were produced. Matrix content ranged from 1% to 5% (w/v), and polymer from 0.75% to 1.25% (w/v). GG hydrogels were used as negative control. Preliminary results demonstrated the possibility to conjugate components from hDFB’s ECM with GG, obtaining stable gels under in vitro conditions for up to 14 days. During this timeframe, protein released from the gels was quantified by Bradford Protein Quantification Assay. A gradual release of protein content occurred within the first 7 days, while the structural integrity of the gels was maintained. Regarding cell studies, 1 x 104 hDFBs were encapsulated within the gels, and its cytocompatibility was assessed by Live/Dead and Phalloidin/DAPI staining. Good cell viability was observed in all formulations, although cytoskeletal staining showed that the content on ECM components determines cell adhesion and morphology. These results confirmed the proposed hypothesis and a ECM-dependent responsiveness of the cells towards the material. Acknowledgments: FCT/MCTES PD/BD/135248/2017; FCT Doctoral Program in Tissue Engineering, Regenerative Medicine and Stem Cells PD/00169/2013; and Consolidator Grant Project “ECM_INK” (ERC-2016-COG-726061).

Conference NameFoReCaST Second Workshop
Date Published2019-07-08
Conference LocationPorto
KeywordsDermal Fibroblasts, Extracellular matrix, Hydrogels
Peer reviewedno

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