Archives of Dermatological Research

, Volume 300, Issue 1, pp 27–35 | Cite as

Inhibition of retinoic acid-induced skin irritation in calorie-restricted mice

  • James Varani
  • Narasimharao Bhagavathula
  • Muhammad Nadeem Aslam
  • Kevin Fay
  • Roscoe L. Warner
  • Andrew Hanosh
  • Adam G. Barron
  • Richard A. Miller
Original Paper


Mice on a calorie-restricted (CR) diet (total calories restricted to 70% of ad libitum; AL) for periods of time ranging from 3 to 18 months were examined for response to topical treatment with all-trans retinoic acid (RA). Daily application of a 0.1% solution of RA to the shaved skin of UM-HET3 mice on an AL diet produced a severe irritation that was evident by day 4, maximal at day 7–8 and still detectable at day 14. Skin irritation was characterized by redness, dryness, flaking and failure of the hair to grow at the treated site. In CR mice, the same treatment produced little detectable irritation. Animals were sacrificed at the end of the retinoid-treatment period (day 7 or day 14) and skin from these animals was examined histologically. In both AL and CR mice, a similar degree of epidermal hyperplasia was observed. Numerous inflammatory cells (mononuclear cells and granulocytes) were present in the skin of both groups. Occasional S100-positive cells (presumably Langerhans cells) were also observed in the epidermis of skin from both groups. S100-positive cells were also observed in the dermis. When skin from CR and AL mice was incubated in organ culture for 3 days (on day 7 after initiation of RA treatment), similar levels of four different pro-inflammatory cytokines were found in the conditioned medium. Soluble type I collagen levels were also similar. In contrast, the level of matrix metalloproteinase-9 was lower in the conditioned medium of skin from CR mice than in conditioned medium from skin cultures of AL mice. Taken together, these studies suggest that CR may provide a way to mitigate the irritation that normally accompanies RA treatment without compromising the beneficial effects of retinoid use. CR appears to exert a protective effect at the target tissue level rather than by a reduction in pro-inflammatory events, per se.


Retinoic acid Skin irritation Calorie restriction Epidermal hyperplasia Inflammation Cytokines  Matrix metalloproteinases 



This study was supported in part by an Older Americans Independence Center Grant (NIH AG024824) from the USPHS.


  1. 1.
    Ashcroft G, Mills SJ, Ashworth JJ (2002) Ageing and wound healing. Biogerentology 3:337–345CrossRefGoogle Scholar
  2. 2.
    Baeuerele PA, Baltimore D (1996) NF-kB: ten years after. Cell 87:13–20CrossRefGoogle Scholar
  3. 3.
    Basak PY, Eroglu E, Altuntas I, Agalar F, Basak K, Sutcu R (2002) Comparison of the effects of tretinoin, adapalene and collagenase in an experimental model of wound healing. Eur J Dermatol 12:145–8PubMedGoogle Scholar
  4. 4.
    Chambon P (1996) A decade of molecular biology of retinoic acid receptors. FASEB J 10:940–954PubMedGoogle Scholar
  5. 5.
    Chapellier B, Mark M, Messaddeq N, Calleja C, Warot X, Brocard J, Gerard C, Li M, Metsger D, Ghyselinck NB, Chambon P (2002) Physiological and retinoid-induced proliferations of epidermal basal keratinocytes are differently controlled. EMBO J 21:3402–3413PubMedCrossRefGoogle Scholar
  6. 6.
    Choi KH, Lee HB, Jeong MY et al. (2002) The role of matrix metalloproteinase-9 and tissue inhibitor of metalloproteinase-1 in crypogenic organizing pneumonia. Chest 121:1478–1485PubMedCrossRefGoogle Scholar
  7. 7.
    Chung HY, Sung B, Jung KJ, Zou Y, Yu BP (2006) The molecular inflammatory process and aging. Antioxid Redox Signal 8:572–581PubMedCrossRefGoogle Scholar
  8. 8.
    Conway JG, Wakefield JA, Brown RH, Marron BE, Sekut L, Stimpson SA, McElroy A, Menius A, Jeffreys JJ, Clark RL, McGeehan GM, Connolly KM (1995) Inhibition of cartilage and bone destruction in adjuvant arthritis in the rat by a matrix metalloproteinase inhibitor. J Exp Med 182:449–457PubMedCrossRefGoogle Scholar
  9. 9.
    Elias PM, Fritsch P, Lampe M, Williams M, Brown B, Nemanic MK, Grayson S (1981) Retinoid effects on epidermal structure, differentiation and permeability. Lab Invest 44:531–540PubMedGoogle Scholar
  10. 10.
    Fisher GJ, Datta SC, Talwar HS, Wang ZQ, Varani J, Kang S, Voorhees JJ (1996) The molecular basis of sun-induced premature skin ageing and retinoid antagonism. Nature (London) 379:335–338CrossRefGoogle Scholar
  11. 11.
    Fisher GJ, Wang ZQ, Datta SC, Varani J, Kang S, Voorhees JJ (1997) Pathophysiology of premature skin aging induced by ultraviolet light. New Engl J Med 337:1419–1428PubMedCrossRefGoogle Scholar
  12. 12.
    Fligiel SEG, Varani J, Datta SH, Kang S, Fisher GJ, Voorhees JJ (2003) Collagen degradation in aged/photoaged skin in vivo and after exposure to MMP-1 in vitro. J Invest Dermatol 120:842–848PubMedCrossRefGoogle Scholar
  13. 13.
    Floyd EE, Jetten AM (1989) Regulation of type I (epidermal) transglutaminase mRNA levels during squamous differentiation: down-regulation by retinoids. Mol Cell Biol 9:4846–4851PubMedGoogle Scholar
  14. 14.
    Griffiths CE, Kang S, Ellis CN, Kim KJ, Finkel LJ, Ortiz-Ferrer LC, White GM, Hamilton TA, Voorhees JJ (1995) Two concentrations of topical tretinoin (retinoic acid) cause similar improvement of photoaging but different degrees of irritation. A double-blind, vehicle-controlled comparison of 0.1% and 0.025% tretinoin cream. Arch Dermatol 131:1037–1044PubMedCrossRefGoogle Scholar
  15. 15.
    Griffiths CEM, Russman G, Majmudar G, Singer RS, Hamilton TA, Voorhees JJ (1993) Restoration of collagen formation in photodamaged human skin by tretinoin (retinoic acid). New Eng J Med 329:530–534PubMedCrossRefGoogle Scholar
  16. 16.
    Halliday GM, McArdle JP, Knight BA, Muller HK (1986) New methodology for assessment of the Langerhans cell network. J Pathol 148:127–134PubMedCrossRefGoogle Scholar
  17. 17.
    Halliwell B, Gutteridge JMC (1985) Free radicals in biology and medicine. Clarenden Press, OxfordGoogle Scholar
  18. 18.
    Heilbronn LK, Jonge de L, Frisard MI, DeLany JP, Larson-Meyer DE, Rood J, Nguyen T, Martin CK, Volaufova J, Most MM, Greenway FL, Smith SR, Deutsch WA, Williamson DA, Ravussin E (2006) Effect of 6-month calorie restriction on biomarkers of longevity, metabolic adaptation, and oxidative stress in overweight individuals: a randomized controlled trial. JAMA 295:1539–1548PubMedCrossRefGoogle Scholar
  19. 19.
    Hyun DH, Emerson SS, Jo DG, Mattson MP, de Cabo R (2006) Calorie restriction up-regulates the plasma membrane redox system in brain cells and suppresses oxidative stress during aging. Proc Natl Acad Sci (USA) 103:1990–1991Google Scholar
  20. 20.
    Jetten AM, George MA, Pettit GR, Herald CL, Rearick JL (1989) Action of phorbol esters, bryostatins and retinoic acid on cholesterol sulfate synthesis: relation to the multistep process of differentiation in human epidermal keratinocytes. J Invest Dermatol 93:108–115PubMedCrossRefGoogle Scholar
  21. 21.
    Kafi R, Kwak HSR, Schumacher WE, Cho S, Hanft VN, Hamilton TA, King AY, Neal JD, Varani J, Fishere GJ, Voorhees JJ, Kang S (2007) Vitamin A (improves) naturally aged skin. Arch Dermatol 143:606–612PubMedCrossRefGoogle Scholar
  22. 22.
    Kahn HJ, Marks A, Thom H, Baumal R (1983) Role of antibody to S100 protein in diagnostic pathology. Am J Clin Pathol 79:341–347PubMedGoogle Scholar
  23. 23.
    Kang S, Duell EA, Fisher GJ, Datta SC, Wang ZQ, Reddy AP, Tavakkol A, Voorhees JJ (1995) Application of retinol to human skin in vivo induces epidermal hyperplasia and cellular retinoid-binding proteins characteristic of retinoic acid but without measurable retinoic acid levels or irritation. J Invest Dermatol 105:549–556PubMedCrossRefGoogle Scholar
  24. 24.
    Kim HJ, Jung KJ, Yu BP, Cho CG, Choi JS, Chung HY (2002) Modulation of redox-sensitive transcription factors by calorie restriction during aging. Mech Ageing Dev 123:1589–1595PubMedCrossRefGoogle Scholar
  25. 25.
    Kim HJ, Jung KJ, Yu BP, Cho CG, Chung HY (2002) Influence of aging and calorie restriction on MAPK activity in rat kidney. Exp Gerontol 37:1041–1053PubMedCrossRefGoogle Scholar
  26. 26.
    Kligman AM, Dogadkina D, Lavker RM (1993) Effects of topical tretinoin on non-sun-exposed protected skin of the elderly. J Am Acad Dermatol 29:25–33PubMedCrossRefGoogle Scholar
  27. 27.
    Kligman LH, Duo CH, Kligman AM (1984) Topical retinoic acid enhances the repair of ultraviolet damaged dermal connective tissue. Connect Tissue Res 12:139–150PubMedCrossRefGoogle Scholar
  28. 28.
    Kligman LH (1986) Effects of all-trans-retinoic acid on the dermis of hairless mice. J Am Acad Dermatol 15:779–785, 884–777PubMedGoogle Scholar
  29. 29.
    Kligman AM, Grove GL, Hirose R, Leyden JJ (1986) Topical tretinoin for photoaged skin. J Am Acad Dermatol 15:836–859PubMedCrossRefGoogle Scholar
  30. 30.
    Laganiere S, Yu BP (1987) Modulation of membrane phospholipids fatty acid composition by age and food restriction. Gerontology 39:7–18Google Scholar
  31. 31.
    Laganiere S, Yu BP (1987) Anti-lipid peroxidation action of food restriction. Biochem Biophys Res Commun 145:1185–1191PubMedCrossRefGoogle Scholar
  32. 32.
    Larson-Meyer DE, Heilbronn LK, Redman LM, Newcomer BR, Frisard MI, Anton S, Smith SR, Alfonso A, Ravussin E (2006) Effect of calorie restriction with or without exercise on insulin sensitivity, beta-cell function, fat cell size, and ectopic lipid in overweight subjects. Diabetes Care 29:1337–1344PubMedCrossRefGoogle Scholar
  33. 33.
    Lateef H, Abatan OI, Aslam MN, Stevens MJ, Varani J (2005) Pretreatment of diabetic rats with all-trans retinoic acid increases healing of subsequently-induced abrasion wounds. Diabetes 54:855–861PubMedCrossRefGoogle Scholar
  34. 34.
    Masoro EJ (2005) Overview of caloric restriction and ageing. Mech Aging Develop 126:913–922CrossRefGoogle Scholar
  35. 35.
    Meeks RG, Zaharevitz D, Chen RF (1981) Membrane effects of retinoids: possible correlation with toxicity. Arch Biochem Biophys 207:141–147PubMedCrossRefGoogle Scholar
  36. 36.
    Merry BJ (2002) Molecular mechanisms linking calorie restriction and longevity. Int J Biochem Cell Biol 34:1340–1354PubMedCrossRefGoogle Scholar
  37. 37.
    Merry BJ (2004) Oxidative stress and mitochondrial function with aging—the effects of calorie restriction. Aging Cell 3:7–12PubMedCrossRefGoogle Scholar
  38. 38.
    Miller RA, Austad S, Burke D, Chrisp C, Dysko R, Galecki A, Jackson A, Monnier V (1999) Exotic mice as models for aging research: polemic and prospectus. Neurobiol Aging 20:217–231PubMedCrossRefGoogle Scholar
  39. 39.
    Miller RA, Chang Y, Galecki AT, Al-Regaiey K, Kopchick JJ, Bartke A (2002) Gene expression patterns in calorically restricted mice: partial overlap with long-lived mutant mice. Mol Endocrinol 16:2657–2666PubMedCrossRefGoogle Scholar
  40. 40.
    Miller RA, Chrisp C (2002) T cell subset patterns that predict resistance to spontaneous lymphoma, mammary adenocarcinoma, and fibrosarcoma in mice. J Immunol 169:1619–1625PubMedGoogle Scholar
  41. 41.
    Peters SL, Mathy MJ, Pfaffendorf M, van Zwieten PA (2000) Reactive oxygen species-induced aortic vasoconstriction and deterioration of functional integrity. Nauyn Schmiedebergs Arch Pharmacol 361:127–133CrossRefGoogle Scholar
  42. 42.
    Popp C, Kligman AM, Stoudemayer TJ (1995) Pretreatment of photoaged forearm skin with topical tretinoin accelerates healing of full-thickness wounds. Brit J Dermatol 132:46–53CrossRefGoogle Scholar
  43. 43.
    Racette SB, Weiss EP, Villareal DT, Arif H, Steger-May K, Schechtman KB, Fontana L, Klein LS, Holloszy JO (2006) One year of caloric restriction in humans: feasibility and effects on body composition and abdominal adipose tissue. J Gerontol A Biol Sci Med Sci 61(9):943–950PubMedGoogle Scholar
  44. 44.
    Suga M, Iyonaga K, Okamoto T et al. (2000) Characteristic elevation of matrix metalloproteinase activity in idiopathic interstitial pneumonias. Am J Respir Crit Care Med 162:1949–1956PubMedGoogle Scholar
  45. 45.
    Talwar HS, Griffiths CEM, Fisher GJ, Hamilton TA, Voorhees JJ (1995) Reduced type I and type III procollagens in photodamaged adult human skin. J Invest Dermatol 105:285–290PubMedCrossRefGoogle Scholar
  46. 46.
    Varani J, Shayevitz J, Perry D, Mitra RS, Nickoloff BJ, Voorhees JJ (1990) Retinoic acid stimulation of human dermal fibroblast proliferation is dependent on suboptimal extracellular Ca2+ concentration. Am J Path 136:1275–1281PubMedGoogle Scholar
  47. 47.
    Varani J, Gibbs DF, Inman DR, Shah B, Fligiel SEG, Voorhees JJ (1991) Inhibition of epithelial cell adhesion by retinoic acid: relationship to reduced extracellular matrix production and alterations in Ca2+ levels. Am J Path 138:887–895PubMedGoogle Scholar
  48. 48.
    Varani J, Burmeister W, Bleavins MR, Johnson JK (1996) All-trans retinoic acid reduces membrane fluidity of dermal fibroblasts: assessment of fluorescence redistribution after photobleaching. Amer J Pathol 148:1307–1312Google Scholar
  49. 49.
    Varani J, Warner RL, Gharaee-Kermani M, Phan SH, Kang S, Chung J, Wang ZQ, Datta SH, Fisher GJ, Voorhees JJ (2000) Vitamin A antagonizes decreased cell growth and elevated collagen-degrading matrix metalloproteinases and stimulates collagen accumulation in naturally aged human skin. J Invest Dermatol 114:480–486PubMedCrossRefGoogle Scholar
  50. 50.
    Varani J, Zeigler M, Dame MK, Kang S, Fisher GJ, Voorhees JJ, Stoll SW, Elder JT (2001) Heparin-binding epidermal growth factor-like growth factor activation of keratinocyte ErbB receptor mediates epidermal hyperplasia, a prominent side effect of retinoid therapy. J Invest Dermatol 1117:1335–1341CrossRefGoogle Scholar
  51. 51.
    Varani J, Fligiel H, Zhang J, Aslam MN, Lu Y, Dehne LA, Keller ET (2003) Separation of retinoid-induced epidermal and dermal thickening from skin irritation. Arch Dermatol Res 295:255–262PubMedCrossRefGoogle Scholar
  52. 52.
    Varani J, Fay K, Perone P (2007) MDI-301: a non-irritating retinoid, induces changes in organ-cultured human skin that underlie repair. Arch Dermatol Res 298:439–448PubMedCrossRefGoogle Scholar
  53. 53.
    Varani J, Perone P, Spahlinger DM, Singer LM, Diegel KL, Bobrowski WF, Dunstan R (2007) Human skin in organ culture and human skin cells (keratinocytes and fibroblasts) in monolayer culture for assessment of chemically-induced skin damage. Toxicol Pathol 35:693–701 PubMedCrossRefGoogle Scholar
  54. 54.
    Weiss JS, Ellis CN, Voorhees JJ (1988) Topical tretinoin improves photoaged skin: a double blind, vehicle-controlled study. JAMA 259:527–532PubMedCrossRefGoogle Scholar
  55. 55.
    Wicke C, Halliday B, Allen D, Roche NS, Scheuenstuhl H, Spencer MM, Roberts AB, Hunt TK (2000) Effects of steroids and retinoids on wound healing. Arch Surgery 135:1265–1270CrossRefGoogle Scholar
  56. 56.
    Wood LC, Elias PM, Calhoun C, Tsai JC, Grunfeld C, Feingold KR (1996) Barrier disruption stimulates interleukin-1a expression and release from a pre-formed pool in murine epidermis. J Invest Dermatol 106:397–403PubMedCrossRefGoogle Scholar
  57. 57.
    Yuen DE, Stratford AF (2004) Vitamin A activation of transforming growth factor-beta1 enhances porcine ileum wound healing in vitro. Pediatr Res 55:935–939PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • James Varani
    • 1
  • Narasimharao Bhagavathula
    • 1
  • Muhammad Nadeem Aslam
    • 1
  • Kevin Fay
    • 1
  • Roscoe L. Warner
    • 1
  • Andrew Hanosh
    • 1
  • Adam G. Barron
    • 1
  • Richard A. Miller
    • 1
    • 2
    • 3
  1. 1.Department of PathologyThe University of MichiganAnn ArborUSA
  2. 2.The Geriatrics CenterUniversity of Michigan Medical SchoolAnn ArborUSA
  3. 3.Department of PathologyAnn Arbor VA Medical CenterAnn ArborUSA

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