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Use of Human Gingival Fibroblasts for Pre-Vascularization Strategies in Oral Tissue Engineering

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

Background:

Cocultures of human gingival fibrobasts (hGF) and endothelial cells could enhance regeneration and repair models as well as improve vascularization limitations in tissue engineering. The aim of this study was to assess if hGF could support formation of stable vessel-like networks.

Methods:

Explant primary hGF were isolated from gum surgical wastes collected from healthy patients with no history of periodontitis. Human umbilical vein endothelial cells (HUVEC) were two-dimensional (2D) and three-dimensional (3D) cocultured in vitro with hGF at a cell ratio of 1:1 and medium of 1:1 of their respective media during at least 31 days. Vessel quantification of HUVEC networks was performed. In order to investigate the pericyte-like properties of hGF, the expression of perivascular markers α-SMA, NG2, CD146 and PDGFR-β was studied using immunocytochemistry and flow cytometry on 2D cultures.

Results:

hGF were able to support a long-lasting HUVEC network at least 31 days, even in the absence of a bioreactor with flow. As observed, HUVEC started to communicate with each other from day 7, constructing a network. Their interconnection increased significantly between day 2 and day 21 and lasted beyond the 31 days of observation. Moreover, we tried to explain the stability of the networks obtained and showed that a small population of hGF in close vicinity of HUVEC networks expressed perivascular markers.

Conclusion:

These findings highlight a new interesting property concerning hGF, accentuating their relevance in tissue engineering and periodontal regeneration. These promising results need to be confirmed using more 3D applications and in vivo testing.

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Acknowledgements

This work was supported by a grant from the Fondation de l’Avenir pour la Recherche Médicale [AP-RM-20-025]. The authors would like to thank Hugo Oliveira (ART BioPrint, Bordeaux) for providing the collagen/hyaluronic acid biomaterial ink for 3D coculture.

Funding

Fondation de l'Avenir pour la Recherche Médicale Appliquée, AP-RM-20-025, Adrien Naveau.

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Authors and Affiliations

  1. Univ. Bordeaux, INSERM, Laboratoire Bioingénierie Tissulaire (BioTis), U1026, CHU Bordeaux, Univ. Bordeaux, 33 076, Bordeaux, France

    Rawen Smirani, Raphaël Devillard & Adrien Naveau

  2. Univ. Bordeaux, CNRS, Chimie et Biologie des Membranes et des Nanoobjets (CBMN), U5248, Univ. Bordeaux, 33600, Pessac, France

    Murielle Rémy

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  1. Rawen Smirani
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Correspondence to Rawen Smirani.

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Ethical statement

Samples were harvested according to the French legislation i.e. under the control of the declaration for conservation and preparation of human body elements for scientific research number DC 2008-412 (French ministry of higher education, research and innovation). Protocols were approved by the institutional committee for the protection of human subjects (Local Ethic Committee from academic hospital CHU de Bordeaux).

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Smirani, R., Rémy, M., Devillard, R. et al. Use of Human Gingival Fibroblasts for Pre-Vascularization Strategies in Oral Tissue Engineering. Tissue Eng Regen Med 19, 525–535 (2022). https://doi.org/10.1007/s13770-021-00415-3

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  • DOI: https://doi.org/10.1007/s13770-021-00415-3

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