Porcine Skin Flow‐Through Diffusion Cell System

Ronald E. Baynes1

1 North Carolina State University, Raleigh, North Carolina
Publication Name:  Current Protocols in Toxicology
Unit Number:  Unit 5.5
DOI:  10.1002/0471140856.tx0505s09
Online Posting Date:  November, 2001
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Abstract

Porcine Skin Flow‐Through Diffusion Cell System (Ronald E. Baynes, North Carolina State University, Raleigh, North Carolina). Porcine skin can be used in a diffusion cell apparatus to study the rate and extent of absorption of topically applied chemicals through the skin. Although the skin of a number of animals can be used in this system, that of the pig most closely approximates human skin anatomically and physiologically.

     
 
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Table of Contents

  • Basic Protocol 1: Operation of the Flow‐through Diffusion Cell System
  • Support Protocol 1: Surgical Preparation of Porcine Skin
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1: Operation of the Flow‐through Diffusion Cell System

  Materials
  • Artificial medium
  • Skin punch biopsy sections (see protocol 2)
  • Bovine serum albumin medium (unit 5.4)
  • Test compound
  • 1% (v/v) soapy solution
  • Sodium hypochlorite
  • Isopropyl alcohol
  • Two‐compartment Teflon flow‐through diffusion cells (Crown Glass)
  • Heating blocks
  • Tygon tubing
  • Brinkmann constant‐temperature circulator (Brinkmann)
  • Peristaltic pump
  • Fraction collector
  • Humidifier
  • Hygrometer and thermometer
  • pH meter
  • 20‐ml scintillation vials
  • Cotton swabs (e.g., Q‐tips)
  • Cellophane tape
  • Magnetic stirrer

Support Protocol 1: Surgical Preparation of Porcine Skin

  Materials
  • 8‐week‐old weanling female Yorkshire pigs (e.g., Looper Farms)
  • Preanesthetic: ketamine hydrochloride (11 mg/kg body weight); xylazine hydrochloride (1.4 mg/kg body weight)
  • Pentobarbitol sodium (100 mg/kg body weight)
  • Physiological saline
  • Gauze
  • Dermatome and blades (e.g., Padgett electric model; optional)
  • Scissors and forceps
  • 16‐mm steel biopsy punch
  • Micrometer
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Figures

Videos

Literature Cited

Literature Cited
   Bronaugh, R. and Collier, S. 1993. In vitro methods for measuring skin permeation. In Skin Permeation (J.L. Zatz, ed.) pp. 93‐111, Allured Publishing, Wheaton, Ill.
   Bronaugh, R.L. and Stewart, R.F. 1984. Methods for in vitro percutaneous absorption studies III: Hydrophobic compounds. J. Pharm. Sci. 73:1255‐1258.
   Bronaugh, R.L. and Stewart, R.F. 1985. Methods for in vitro percutaneous absorption studies IV: The flow‐through diffusion cell. J. Pharm. Sci. 74:64‐67.
   Chang, S.K. and Riviere, J.E. 1991. Percutaneous absorption of parathion in vitro in porcine skin: Effect of dose, temperature, humidity, and perfusate composition on absorptive flux . Fundam. Appl. Toxicol. 17:494‐504.
   Crutcher, W. and Maibach, H.I. 1969. The effect of percutaneous rate on in vitro percutaneous penetration. J. Investig. Dermatol. 53:264‐269.
   Grummer, C. and Maibach, H.I. 1991. Diffusion cell design. In In Vitro Percutaneous Absorption: Principles, Fundamentals and Applications (R.L. Bronaugh and H.I. Maibach, eds.) pp. 7‐16, CRC Press, Boca Raton, Fla.
   Monteiro‐Riviere, N.A. 1991. Comparative anatomy, physiology, and biochemistry of mammalian skin. In Dermal and Ocular Toxicology: Fundamentals and Methods (D.W. Hobson, ed.) pp. 3‐71, CRC Press, Boca Raton, Fla.
   Reifenrath, W.G., Hawkins, G.S., and Kurtz, M.S. 1991. Percutaneous penetration and skin retention of topically applied compounds: An in vitro–in vivo study. J. Pharm. Sci. 80:526‐532.
   Scott, R.C. and Ramsey, J.D. 1987. Comparison of the in vivo and in vitro percutaneous absorption of a lipophilic molecule (Cypermethrin, a pyrethrin insecticide). J. Investig. Dermatol. 89:142‐146.
   Scott, R.C., Corrigan, M.A., Smith, F., and Mason, H. 1991. The influence of skin structure on permeability: An intersite and interspecies comparison with hydrophilic penetrants. J. Investig.Dermatol. 96:921‐925.
Key References
   Bronaugh and Stewart, 1985. See above.
   This paper describes the use and advantages of flow‐through diffusion cells in assessing in vitro dermal absorption of chemicals.
   Chang and Riviere, 1991. See above.
   This paper describes the use of flow‐through diffusion cells in an environmentally controlled system in order to assess environmental effects on in vitro dermal absorption of chemicals.
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