Enhancement of gap junctional intercellular communication of normal human dermal fibroblasts cultured on polystyrene dishes grafted with poly-N-isopropylacrylamide.
Technology developed to allow recovery of cells without enzyme treatment, involving a dish grafted with a thermoreactive polymer gel of poly-N-isopropylacrylamide (PIPAAm), was found to significantly enhance gap junctional intercellular communication (GJIC) in normal human dermal fibroblasts (NHDF cells). NHDF cells were cultured for 4 days on PIPAAm-grafted dishes irradiated with various doses of electron beams, and GJIC was assayed by the scrape-loading dye transfer method. The area of dye transfer was greater in the PIPAAm-grafted dishes than in the control culture dishes, indicating that the PIPAAm-grafted dishes enhanced the GJIC of NHDF cells. Connexin-43 (Cx43) expression was analyzed because Cx43 is considered to be a main component of the gap junctional channel. PIPAAm-grafted dishes irradiated with 100, 250, or 500 kGy of electron beams showed significantly enhanced expression of Cx43-NP, Cx43-P1, and especially Cx43-P2. Enhanced expression of Cx43-P2, a functional transmembrane protein, may be related to the promotion of GJIC. These results suggest that the PIPAAm-grafted dish not only enables the enzyme-free recovery of a cell monolayer for use in the construction of a three-dimensional artificial tissue, but also significantly contributes to the enhancement of GJIC, which may partly promote tissue strength on the surface of the PIPAAm-grafted dish.
Nagira
Tsutomu
T
Division of Medical Devices, National Institute of Health Sciences, Tokyo, Japan.
Matthew
Susan Bijoo
SB
Yamakoshi
Yoko
Y
Tsuchiya
Toshie
T
D000180:Acrylic Resins; D001672:Biocompatible Materials; D015153:Blotting, Western; D002450:Cell Communication / Q000502:physiology*; D018929:Cell Culture Techniques; D002460:Cell Line; D018031:Connexin 43 / Q000032:analysis / Q000378:metabolism; D004583:Electrons; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology* / Q000528:radiation effects*; D017629:Gap Junctions / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D011137:Polystyrenes; D012867:Skin / Q000166:cytology; D023822:Tissue Engineering
D000180:Acrylic Resins; D001672:Biocompatible Materials; D018031:Connexin 43; D011137:Polystyrenes; C052970:poly-N-isopropylacrylamide