Ann Dermatol.  2012 May;24(2):181-188.

The Effect of Adipose-Derived Stem Cell-Cultured Media on Oxazolone Treated Atopic Dermatitis-Like Murine Model

• Lee HJ
• Jung M
• Kim JH
• Yoon NY
• Choi EH

Abstract

BACKGROUND
A stem cell is an undifferentiated cell that has the potential for self-renewal and differentiation. Adipose-derived stem cells (ADSCs) have advantages in accessibility and abundance compared to other kinds of stem cells and produce many growth factors and hormones.
OBJECTIVE
We investigated whether ADSC cultured media could be used as a therapy for atopic dermatitis.
METHODS
ADSC cultured media was topically applied twice daily for 5 days to oxazolone-treated atopic dermatitis-like hairless mice.
RESULTS
Topical application of ADSC cultured media improved the epidermal permeability barrier and keratinocyte differentiation, and restored the predominant Th2 phenotype when compared to vehicle. ADSC cultured media-treated epidermis also showed an increase in the expression of antimicrobial peptides cathelin-related antimicrobial peptide, mouse beta-defensein 3.
CONCLUSION
Topical ADSC cultured media could be useful in the treatment of atopic dermatitis.

Keyword

Adipose-derived stem cell; Atopic dermatitis; Oxazolone

MeSH Terms

Animals
Cathelicidins
Dermatitis, Atopic
Epidermis
Intercellular Signaling Peptides and Proteins
Keratinocytes
Mice
Oxazolone
Peptides
Permeability
Phenotype
Stem Cells
Cathelicidins
Intercellular Signaling Peptides and Proteins
Oxazolone
Peptides

Figure

• 

  Fig. 1 Methods of multiple oxazolone (Ox) challenges and topical administration of adipose-derived stem cell cultured media.

• 

  Fig. 2 (A) Basal TEWL (gm-2h-1) was checked after the fifth challenge of oxazolone (MM-pre, AM-pre, FM-pre, S-pre). The AM group and S group showed significant decreases in TEWL after treatment (C-post, MM-post, AM-post, FM-post and S-post). (B) SC hydration after treatment. AM, FM, and S groups showed significant improvement of SC hydration. Data are expressed as mean±standard deviation; n=6 in MM, AM, and FM group; n=4 in C group; n=3 in S group. TEWL: transepidermal water loss, SC: stratum corneum, C: control group, MM: minimal media applied group, AM: adipose-derived stem cell cultured media applied group, FM: fibroblast cultured media applied group, S: steroid applied group.

• 

  Fig. 3 Gross lesions of pre- and post-treatment with media in the atopic dermatitis model. The AM group and S group showed an improvement in skin lesions grossly. C: control group, MM: minimal media applied group, AM: adipose-derived stem cell cultured media applied group, FM: fibroblast cultured media applied group, S: steroid applied group.

• 

  Fig. 4 Th2 immunophenotype (CRTH2) and keratinocyte differentiation (filaggrin, involucrin, and loricrin) after treatment. Immunohistochemical staining was carried out with diaminobenzidine and primary antibodies as indicated. Merged images with propidium iodide secondary staining in 5 mm-thick frozen sections. C: control group, MM: minimal media applied group, AM: adipose-derived stem cell cultured media applied group, FM: fibroblast cultured media applied group, S: steroid applied group (Mag bars=2 mm).

• 

  Fig. 5 Topical application of adipose-derived stem cell cultured media enhanced the epidermal expression of antimicrobial peptides such as mBD3 and CRAMP. Immunohistochemical staining was carried out with diaminobenzidine and primary antibodies as indicated. mBD3: mouse beta-defensin 3, CRAMP: cathelin-related antimicrobial peptide, C: control group, MM: minimal media applied group, AM: adipose-derived stem cell cultured media applied group, FM: fibroblast cultured media applied group, S: steroid applied group (Mag bars=2 mm).

• 

  Fig. 6 Topical application of adipose-derived stem cell cultured media increased the protein and mRNA levels of mBD3 and CRAMP compared with other treatments in Western blots (A) and for mRNA of mBD3 and CRAMP compared with vehicle in real-time reverse transcriptase-polymerase chain reaction (B). This is compatible with immunohistochemical staining results. Data are expressed as mean±standard deviation; n=6 in MM, AM and FM group; n=4 in C group; n=3 in S group. CRAMP: cathelin-related antimicrobial peptide; mBD3: mouse beta-defensin 3, C: control group, MM: minimal media applied group, AM: adipose-derived stem cell cultured media applied group, FM: fibroblast cultured media applied group, S: steroid applied group.

Reference

1. O'Regan GM, Irvine AD. The role of filaggrin in the atopic diathesis. Clin Exp Allergy. 2010. 40:965–972.

2. Kern S, Eichler H, Stoeve J, Klüter H, Bieback K. Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells. 2006. 24:1294–1301.

3. Ripoll CB, Flaat M, Klopf-Eiermann J, Fisher-Perkins JM, Trygg CB, Scruggs BA, et al. Mesenchymal lineage stem cells have pronounced anti-inflammatory effects in the twitcher mouse model of Krabbe's disease. Stem Cells. 2011. 29:67–77.

4. Razmkhah M, Jaberipour M, Erfani N, Habibagahi M, Talei AR, Ghaderi A. Adipose derived stem cells (ASCs) isolated from breast cancer tissue express IL-4, IL-10 and TGF-β1 and upregulate expression of regulatory molecules on T cells: do they protect breast cancer cells from the immune response? Cell Immunol. 2011. 266:116–122.

5. Crop MJ, Baan CC, Korevaar SS, Ijzermans JN, Weimar W, Hoogduijn MJ. Human adipose tissue-derived mesenchymal stem cells induce explosive T-cell proliferation. Stem Cells Dev. 2010. 19:1843–1853.

6. Man MQ, Hatano Y, Lee SH, Man M, Chang S, Feingold KR, et al. Characterization of a hapten-induced, murine model with multiple features of atopic dermatitis: structural, immunologic, and biochemical changes following single versus multiple oxazolone challenges. J Invest Dermatol. 2008. 128:79–86.

7. Aberg KM, Man MQ, Gallo RL, Ganz T, Crumrine D, Brown BE, et al. Co-regulation and interdependence of the mammalian epidermal permeability and antimicrobial barriers. J Invest Dermatol. 2008. 128:917–925.

8. Choi EH, Brown BE, Crumrine D, Chang S, Man MQ, Elias PM, et al. Mechanisms by which psychologic stress alters cutaneous permeability barrier homeostasis and stratum corneum integrity. J Invest Dermatol. 2005. 124:587–595.

9. Ong PY, Ohtake T, Brandt C, Strickland I, Boguniewicz M, Ganz T, et al. Endogenous antimicrobial peptides and skin infections in atopic dermatitis. N Engl J Med. 2002. 347:1151–1160.

10. Aberg KM, Radek KA, Choi EH, Kim DK, Demerjian M, Hupe M, et al. Psychological stress downregulates epidermal antimicrobial peptide expression and increases severity of cutaneous infections in mice. J Clin Invest. 2007. 117:3339–3349.

11. Meyer GP, Wollert KC, Drexler H. Stem cell therapy: a new perspective in the treatment of patients with acute myocardial infarction. Eur J Med Res. 2006. 11:439–446.

12. Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, et al. Isolation of putative progenitor endothelial cells for angiogenesis. Science. 1997. 275:964–967.

13. Elias PM. Therapeutic Implications of a Barrier-based Pathogenesis of Atopic Dermatitis. Ann Dermatol. 2010. 22:245–254.

14. Lai Y, Gallo RL. AMPed up immunity: how antimicrobial peptides have multiple roles in immune defense. Trends Immunol. 2009. 30:131–141.

15. Hong SP, Oh Y, Jung M, Lee S, Jeon H, Cho MY, et al. Topical calcitriol restores the impairment of epidermal permeability and antimicrobial barriers induced by corticosteroids. Br J Dermatol. 2010. 162:1251–1260.

16. Krasnodembskaya A, Song Y, Fang X, Gupta N, Serikov V, Lee JW, et al. Antibacterial effect of human mesenchymal stem cells is mediated in part from secretion of the antimicrobial peptide LL-37. Stem Cells. 2010. 28:2229–2238.