Increase in gap-junctional intercellular communications (GJIC) of normal human dermal fibroblasts (NHDF) on surfaces coated with high-molecular-weight hyaluronic acid (HMW HA).
Normal human dermal fibroblast (NHDF) cells were used to detect differences in gap-junctional intercellular communication (GJIC) by hyaluronic acid (HA), a linear polymer built from repeating disaccharide units that consist of N-acetyl-D-glucosamine (GlcNa) and D-glucuronic acid (GlcA) linked by a beta 1-4 glycosidic bond. The NHDF cells were cultured with different molecular weights (MW) of HA for 4 days. The rates of cell attachment in dishes coated with high-molecular-weight (HMW; 310 kDa or 800 kDa) HA at 2 mg/dish were significantly reduced at an early time point compared with low-molecular-weight (LMW; 4.8 kDa or 48 kDa) HA with the same coating amounts. HA-coated surfaces were observed by atomic force microscopy (AFM) under air and showed that HA molecules ran parallel in the dish coated with LMW HA and had an aggregated island structure in the dish coated with HMW HA surfaces. The cell functions of GJIC were assayed by a scrape-loading dye transfer (SLDT) method using a dye solution of Lucifer yellow. Promotion of the dye transfer was clearly obtained in the cell monolayer grown on the surface coated with HMW HA. These results suggest that HMW HA promotes the function of GJIC in NHDF cells. In contrast, when HMW HA was added to the monolayer of NHDF cells, the functions of GJIC clearly were lowered in comparison with the cells grown in the control dish or with those grown on the surface of HMW HA. Therefore it is concluded that the MW size of HA and its application method are important factors for generating biocompatible tissue-engineered products because of the manner in which the GJIC participates in cell differentiation and cell growth rate.
Park
Jeong Ung
JU
Division of Medical Devices, National Institute of Health Sciences, Kamiyoga, 1-18-1, Setagaya-ku, Tokyo, 158-8501, Japan.
Tsuchiya
Toshie
T
D002448:Cell Adhesion / Q000502:physiology; D002450:Cell Communication / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D020099:Coated Materials, Biocompatible; D020405:Dermis / Q000166:cytology* / Q000502:physiology; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D005456:Fluorescent Dyes; D017629:Gap Junctions / Q000502:physiology*; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry; D007546:Isoquinolines; D018625:Microscopy, Atomic Force; D008970:Molecular Weight
D020099:Coated Materials, Biocompatible; D005456:Fluorescent Dyes; D007546:Isoquinolines; D006820:Hyaluronic Acid; C017475:lucifer yellow