title: Effect of the HIV protease inhibitor amprenavir on the growth and differentiation of primary gingival epithelium.
abstract: BACKGROUND
  HIV-positive patients taking antiretroviral drugs, including protease inhibitors, have shown a significant increase in the development of oral complications; these complications are a major health issue for affected patients. The effect of these drugs on oral epithelium growth and differentiation is presently unknown. In this study, we explore for the first time the effect of the HIV protease inhibitor amprenavir on gingival epithelium growth and differentiation.
  
  
  METHODS
  Organotypic (raft) cultures of gingival keratinocytes were established and the raft cultures treated with a range of amprenavir concentrations. Haematoxylin and eosin staining was performed to examine the effect of amprenavir on gingival epithelium growth and stratification. The raft cultures were also immunohistochemically analysed to determine the effect of amprenavir on the expression of key differentiation and proliferation markers, including cytokeratins, proliferating cell nuclear antigen (PCNA) and cyclin A.
  
  
  RESULTS
  Amprenavir severely inhibited the growth of gingival epithelium when the drug was present throughout the growth period of the tissue. When the drug was added at day 8, amprenavir treatment altered the proliferation and differentiation of gingival keratinocytes. Expression of the cytokeratins 5, 14, 6 and 10, PCNA and cyclin A was increased; their expression pattern was also altered over time in treated rafts. Biochemically, the tissue exhibited characteristics of increased proliferation in the suprabasal layers of amprenavir-treated tissue.
  
  
  CONCLUSIONS
  Our results suggest that amprenavir treatment deregulates the cell cycle/proliferation and differentiation pathways, resulting in abnormal epithelial repair and proliferation. Our system could be developed as a potential model for studying the effects of highly active antiretroviral therapy in vitro.
authors.affiliation: Department of Microbiology and Immunology, Pennsylvania State University College of Medicine, Hershey, PA, USA.
authors.forename: Mohd
authors.initials: M
authors.lastname: Israr
authors.forename: Danielle
authors.initials: D
authors.lastname: Mitchell
authors.forename: Samina
authors.initials: S
authors.lastname: Alam
authors.forename: Donald
authors.initials: D
authors.lastname: Dinello
authors.forename: Joseph J
authors.initials: JJ
authors.lastname: Kishel
authors.forename: Craig
authors.initials: C
authors.lastname: Meyers
mesh_terms: D016475:3T3 Cells; D000818:Animals; D023241:Antiretroviral Therapy, Highly Active; D002219:Carbamates / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D018920:Coculture Techniques; D004848:Epithelium / Q000187:drug effects*; D005347:Fibroblasts; D005663:Furans; D005881:Gingiva / Q000166:cytology / Q000187:drug effects*; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D006801:Humans; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects; D051379:Mice; D009928:Organ Specificity; D013449:Sulfonamides / Q000494:pharmacology*
chemical_list: D002219:Carbamates; D005663:Furans; D017320:HIV Protease Inhibitors; D013449:Sulfonamides; C095108:amprenavir
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