Two sensitive in vitro monitors of chemical toxicity to human and animal skin (in short-term organ culture): I. Paranuclear vacuolization in glycol methacrylate tissue sections II. Interference with [14c]leucine incorporation

K. Gregory Moore, Brian H. Schofield, Kazuwki Higuchi, Akira Kajiki, Kin Wing Au, Peggy J. Pula, David P. Bassett, Arthur M. Dannenberg

Research output: Contribution to journalArticle

Abstract

Two sensitive in vitro tests have been developed to determine the toxicity of chemicals applied to full-thickness skin explants from fresh human surgical specimens and sacrificed guinea pigs and rabbits. Before incubation, dinitrochlorobenzene (DNCB), sulfur mustard, or resorcinol was topically applied to the exposed top surface of each explant. The explants were then organ-cultured for 24 hr in shallow Petri dishes. Toxicity to skin was assessed by two methods. The first was histologic and morphometric. Paranuclear vacuoles in the basal epidermal cells were counted microscopically in tissue sections prepared from the cultured ex-plants. The number of these vacuoles was found to increase with the toxicant concentration, in the range from low to full toxicity. The DNCB (0.5% caused fewer vacuoles in guinea pigs than in the other two species. Resorcinol (50% caused more vacuoles in rabbits than in the other two species. Control skin explants (normal skin and skin exposed only to diluents) contained very few paranuclear vacuoles. Examination of these vacuoles under the electron microscope suggests they might be blebs in the cell's nuclear membrane. The second method to assess skin toxicity was biochemical and functional. The toxicant-exposed skin explants were cultured in the presence of [14C]leucine for 24 hr, along with unexposed explants and with explants exposed to the diluents. The incorporation of [14C]leucine into the proteins of the skin explants was found to decrease as above with the toxicant concentration. These tests for in vitro toxicity to skin proved to be the most sensitive and reliable among the various histochemical and biochemical tests for lysosomal and oxidative enzymes that we performed.

Original languageEnglish (US)
Pages (from-to)285-302
Number of pages18
JournalCutaneous and Ocular Toxicology
Volume5
Issue number4
DOIs
StatePublished - 1986

Fingerprint

Organ Culture Techniques
Leucine
Toxicity
Skin
Animals
Tissue
Vacuoles
Dinitrochlorobenzene
Guinea Pigs
Mustard Gas
Rabbits
hydroxyethyl methacrylate
In Vitro Techniques
Nuclear Envelope
Blister
Electron microscopes
Cell Membrane
Electrons
Membranes
Enzymes

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis
  • Ophthalmology

Cite this

Two sensitive in vitro monitors of chemical toxicity to human and animal skin (in short-term organ culture) : I. Paranuclear vacuolization in glycol methacrylate tissue sections II. Interference with [14c]leucine incorporation. / Moore, K. Gregory; Schofield, Brian H.; Higuchi, Kazuwki; Kajiki, Akira; Au, Kin Wing; Pula, Peggy J.; Bassett, David P.; Dannenberg, Arthur M.

In: Cutaneous and Ocular Toxicology, Vol. 5, No. 4, 1986, p. 285-302.

Research output: Contribution to journalArticle

Moore, K. Gregory ; Schofield, Brian H. ; Higuchi, Kazuwki ; Kajiki, Akira ; Au, Kin Wing ; Pula, Peggy J. ; Bassett, David P. ; Dannenberg, Arthur M. / Two sensitive in vitro monitors of chemical toxicity to human and animal skin (in short-term organ culture) : I. Paranuclear vacuolization in glycol methacrylate tissue sections II. Interference with [14c]leucine incorporation. In: Cutaneous and Ocular Toxicology. 1986 ; Vol. 5, No. 4. pp. 285-302.
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abstract = "Two sensitive in vitro tests have been developed to determine the toxicity of chemicals applied to full-thickness skin explants from fresh human surgical specimens and sacrificed guinea pigs and rabbits. Before incubation, dinitrochlorobenzene (DNCB), sulfur mustard, or resorcinol was topically applied to the exposed top surface of each explant. The explants were then organ-cultured for 24 hr in shallow Petri dishes. Toxicity to skin was assessed by two methods. The first was histologic and morphometric. Paranuclear vacuoles in the basal epidermal cells were counted microscopically in tissue sections prepared from the cultured ex-plants. The number of these vacuoles was found to increase with the toxicant concentration, in the range from low to full toxicity. The DNCB (0.5{\%} caused fewer vacuoles in guinea pigs than in the other two species. Resorcinol (50{\%} caused more vacuoles in rabbits than in the other two species. Control skin explants (normal skin and skin exposed only to diluents) contained very few paranuclear vacuoles. Examination of these vacuoles under the electron microscope suggests they might be blebs in the cell's nuclear membrane. The second method to assess skin toxicity was biochemical and functional. The toxicant-exposed skin explants were cultured in the presence of [14C]leucine for 24 hr, along with unexposed explants and with explants exposed to the diluents. The incorporation of [14C]leucine into the proteins of the skin explants was found to decrease as above with the toxicant concentration. These tests for in vitro toxicity to skin proved to be the most sensitive and reliable among the various histochemical and biochemical tests for lysosomal and oxidative enzymes that we performed.",
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AU - Au, Kin Wing

AU - Pula, Peggy J.

AU - Bassett, David P.

AU - Dannenberg, Arthur M.

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