Interstitial fluid flow-induced growth potential and hyaluronan synthesis of fibroblasts in a fibroblast-populated stretched collagen gel culture

• Published in: Biochimica et biophysica acta. General subjects Vol. 1861; no. 9; pp. 2261 - 2273
• Main Authors: Saito, Natsumi, Adachi, Hiroaki, Tanaka, Hiroshi, Nakata, Satoru, Kawada, Norifumi, Oofusa, Ken, Yoshizato, Katsutoshi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.09.2017
• Subjects: 3-D collagen culture; adenosine; biochemical pathways; Cell Proliferation; Cells, Cultured; Collagen; dermis; Extracellular Fluid - physiology; extracellular matrix; Extracellular Matrix - metabolism; Fibroblast biology; fibroblasts; Fibroblasts - physiology; Gels; genes; Humans; Hyaluronan metabolism; hyaluronan synthase; hyaluronic acid; Hyaluronic Acid - biosynthesis; Interstitial fluid flow; metalloproteinases; phenotype; Tensile strength; Tissue tension; PCNA; Interstitial fluid flow; (H)DFs; Tissue tension; PFA; SC; DMEM; DMEM/S; DW; FBS; IF; CGC; DAB; H&E; MT; qRT-PCR; MF(C); 2-D; CF(s); Hyaluronan metabolism; ECM; Tensile strength; Fibroblast biology; HA; 3-D collagen culture; ATP; ELISA
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2017.06.019

Tensioned collagen gels with dermal fibroblasts (DFs) as a dermis model are usually utilized in a static culture (SC) that lacks medium flowing. To make the model closer to its in vivo state, we created a device to perfuse the model with media flowing at a physiological velocity and examined the effects of medium flow (MF) on the cultures. We constructed a medium perfusion device for human DF-embedded stretched collagen gels (human dermis model), exposed the model to media that flows upwardly at ~1mL/day, and examined water retention of the gels, cells' growth ability, metabolic activity, expression profiles of nine extracellular matrix (ECM)-related genes. The obtained data were compared with those from the model in SC. MF increases the gels' water retention and cells' growth potential but had little effect on their metabolic activities. MF robustly enhanced hyaluronan synthase 2 (HAS2) and matrix metalloprotease 1 (MMP1) gene expressions but not of the other genes (MMP2, HYAL1, HYAL2, HYAL3, COL1A1, COL3A1, and CD44). MF significantly increased the amounts of cellular hyaluronan and adenosine triphosphate. The MF at a physiological speed significantly influences the nature of ECMs and their resident fibroblasts and remodels ECMs by regulating hyaluronan metabolism. Fibroblasts in tensioned collagen gels altered their phenotypes in a MF rate-dependent manner. Collagen gel culture with tension and MF could be utilized as an appropriate in vitro model of interstitial connective tissues to evaluate the pathophysiological significance of mechanosignals generated by fluid flow and cellular/extracellular tension. •A tensioned collagen gel culture system with flowing medium was developed.•Medium flow stimuli increased gels' water retention and cells' growth potential.•Medium flow stimuli prominently increased synthesis and secretion of hyaluronan.•The developed culture system could be a faithful soft interstitial connective tissue model.