title: Biological responses of three-dimensional cultured fibroblasts by sustained compressive loading include apoptosis and survival activity.
abstract: Pressure ulcers are characterized by chronicity, which results in delayed wound healing due to pressure. Early intervention for preventing delayed healing due to pressure requires a prediction method. However, no study has reported the prediction of delayed healing due to pressure. Therefore, this study focused on biological response-based molecular markers for the establishment of an assessment technology to predict delayed healing due to pressure. We tested the hypothesis that sustained compressive loading applied to three dimensional cultured fibroblasts leads to upregulation of heat shock proteins (HSPs), CD44, hyaluronan synthase 2 (HAS2), and cyclooxygenase 2 (COX2) along with apoptosis via disruption of adhesion. First, sustained compressive loading was applied to fibroblast-seeded collagen sponges. Following this, collagen sponge samples and culture supernatants were collected for apoptosis and proliferation assays, gene expression analysis, immunocytochemistry, and quantification of secreted substances induced by upregulation of mRNA and protein level. Compared to the control, the compressed samples demonstrated that apoptosis was induced in a time- and load- dependent manner; vinculin and stress fiber were scarce; HSP90α, CD44, HAS2, and COX2 expression was upregulated; and the concentrations of HSP90α, hyaluronan (HA), and prostaglandin E2 (PGE2) were increased. In addition, the gene expression of antiapoptotic Bcl2 was significantly increased in the compressed samples compared to the control. These results suggest that compressive loading induces not only apoptosis but also survival activity. These observations support that HSP90α, HA, and, PGE2 could be potential molecular markers for prediction of delayed wound healing due to pressure.
authors.affiliation: Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Toshiki
authors.initials: T
authors.lastname: Kanazawa
authors.affiliation: Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Gojiro
authors.initials: G
authors.lastname: Nakagami
authors.affiliation: Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Takeo
authors.initials: T
authors.lastname: Minematsu
authors.affiliation: Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan; Department of Nutritional Sciences, Faculty of Applied Bioscience, Tokyo University of Agriculture, Tokyo, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Yamane
authors.affiliation: Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Lijuan
authors.initials: L
authors.lastname: Huang
authors.affiliation: Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Mugita
authors.affiliation: Department of Life Support Technology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Noguchi
authors.affiliation: Department of Life Support Technology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Taketoshi
authors.initials: T
authors.lastname: Mori
authors.affiliation: Department of Gerontological Nursing/Wound Care Management, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
authors.forename: Hiromi
authors.initials: H
authors.lastname: Sanada
mesh_terms: D000818:Animals; D017209:Apoptosis; D018929:Cell Culture Techniques; D002460:Cell Line; D019245:Compressive Strength; D051546:Cyclooxygenase 2 / Q000096:biosynthesis; D015232:Dinoprostone / Q000096:biosynthesis; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000096:biosynthesis; D018841:HSP90 Heat-Shock Proteins / Q000096:biosynthesis; D018960:Hyaluronan Receptors / Q000096:biosynthesis; D000076002:Hyaluronan Synthases; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000096:biosynthesis; D051381:Rats; D013314:Stress, Mechanical
chemical_list: D018841:HSP90 Heat-Shock Proteins; C588351:Hsp90aa1 protein, rat; D018960:Hyaluronan Receptors; D019253:Proto-Oncogene Proteins c-bcl-2; D051546:Cyclooxygenase 2; C489691:Ptgs2 protein, rat; D014453:Glucuronosyltransferase; C491097:Has2 protein, rat; D000076002:Hyaluronan Synthases; D015232:Dinoprostone
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