Rawen Smirani1, Murielle Rémy2, Raphaël Devillard3, Adrien Naveau3. 1. Univ. Bordeaux, INSERM, Laboratoire Bioingénierie Tissulaire (BioTis), U1026, CHU Bordeaux, Univ. Bordeaux, 33 076, Bordeaux, France. rawen.smirani@u-bordeaux.fr. 2. Univ. Bordeaux, CNRS, Chimie et Biologie des Membranes et des Nanoobjets (CBMN), U5248, Univ. Bordeaux, 33600, Pessac, France. 3. Univ. Bordeaux, INSERM, Laboratoire Bioingénierie Tissulaire (BioTis), U1026, CHU Bordeaux, Univ. Bordeaux, 33 076, Bordeaux, France.
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.
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.
Authors: Hannu Larjava; Colin Wiebe; Corrie Gallant-Behm; David A Hart; Jyrki Heino; Lari Häkkinen Journal: J Can Dent Assoc Date: 2011 Impact factor: 1.316
Authors: Anna Woloszyk; Johanna Buschmann; Conny Waschkies; Bernd Stadlinger; Thimios A Mitsiadis Journal: Front Physiol Date: 2016-04-19 Impact factor: 4.566