Role of phenytoin in wound healing: microarray analysis of early transcriptional responses in human dermal fibroblasts.

Wound healing is a complex process involving a number of related genes and receptors. Using cDNA microarrays, we explored the global gene expression profile of phenytoin (20microg/ml) induced changes to human dermal fibroblasts. Microarray data analysis revealed approximately 1500 genes were differentially expressed by 2.5-fold. At 3, 6, 12, and 24h, the transcripts of the major growth factors involved in wound healing and their receptors were increased. This was further confirmed by RT-PCR. Genes encoding other proteins with roles in signal transduction (NFkappaB), extracellular matrix (MMP1) including type I collagen, fibronectin, and laminin were strongly induced at 6h and onwards. Genes involved in cell cycle regulation (CCND1 and CDKN1A) were down-regulated consistent with our finding that phenytoin per se did not have cell proliferation activity. Notably, phenytoin accelerates the autocrine and paracrine activity of growth factors by up-regulating the related receptors.

[1]  J. Rubin,et al.  Keratinocyte growth factor: an androgen-regulated mediator of stromal-epithelial interactions in the prostate , 2004, World Journal of Urology.

[2]  S. Werner,et al.  Large induction of keratinocyte growth factor expression by serum growth factors and pro-inflammatory cytokines in cultured fibroblasts. , 1994, Oncogene.

[3]  D. Danilenko,et al.  Stimulation of all epithelial elements during skin regeneration by keratinocyte growth factor , 1994, The Journal of experimental medicine.

[4]  A. Gottlieb,et al.  Human keratinocyte growth factor effects in a porcine model of epidermal wound healing , 1993, The Journal of experimental medicine.

[5]  R. Brown,et al.  Role of phenytoin in wound healing--a wound pharmacology perspective. , 1999, Biochemical pharmacology.

[6]  N. Schwartz,et al.  The effects of phenytoin (5,5-diphenylhydantoin) on human gingival fibroblasts in culture. , 1987, Journal of periodontal research.

[7]  M. Fini,et al.  Regulation of Matrix Metalloproteinase Gene Expression , 1998 .

[8]  I. Olsen,et al.  Up-regulation of keratinocyte growth factor and receptor: a possible mechanism of action of phenytoin in wound healing. , 2001, Biochemical and biophysical research communications.

[9]  S. Werner,et al.  Suppression of keratinocyte growth factor expression by glucocorticoids in vitro and during wound healing. , 1995, The Journal of investigative dermatology.

[10]  Richard A.F. Clark,et al.  The Molecular and Cellular Biology of Wound Repair , 2012, Springer US.

[11]  R. Galiano,et al.  Keratinocyte growth factor induces granulation tissue in ischemic dermal wounds. Importance of epithelial-mesenchymal cell interactions. , 1996, Archives of surgery.

[12]  D. Botstein,et al.  The transcriptional program in the response of human fibroblasts to serum. , 1999, Science.

[13]  M J Banda,et al.  Large induction of keratinocyte growth factor expression in the dermis during wound healing. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[14]  H. Nagase,et al.  MMPs Are IGFBP‐Degrading Proteinases: Implications for Cell Proliferation and Tissue Growth , 1999, Annals of the New York Academy of Sciences.

[15]  William C. Parks,et al.  The Activity of Collagenase-1 Is Required for Keratinocyte Migration on a Type I Collagen Matrix , 1997, The Journal of cell biology.

[16]  Y. Yaari,et al.  Phenytoin: Mechanisms of its anticonvulsant action , 1986, Annals of neurology.

[17]  Paul Martin,et al.  Wound Healing--Aiming for Perfect Skin Regeneration , 1997, Science.

[18]  D. Greenhalgh.  Wound healing and diabetes mellitus. , 2003, Clinics in plastic surgery.

[19]  Mukul,et al.  COMPARISON OF TOPICAL PHENYTOIN WITH NORMAL SALINE IN THE TREATMENT OF CHRONIC TROPHIC ULCERS IN LEPROSY , 1993, International journal of dermatology.

[20]  B. Fingleton,et al.  Matrix metalloproteinases: biologic activity and clinical implications. , 2000, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[21]  M. Leader,et al.  Diphenylhydantoin sodium promotes early and marked angiogenesis and results in increased collagen deposition and tensile strength in healing wounds. , 1998, Surgery.

[22]  W. Bottomley,et al.  On the mechanism of drug-induced gingival hyperplasia. , 1991, Journal of oral pathology & medicine : official publication of the International Association of Oral Pathologists and the American Academy of Oral Pathology.

[23]  J. O'Connell,et al.  Reciprocated matrix metalloproteinase activation: a process performed by interstitial collagenase and progelatinase A. , 1994, Biochemistry.

[24]  D. Salomon,et al.  Topical phenytoin accelerates healing in epidermolysis bullosa simplex. , 1995, Dermatology.

[25]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.