PADMA 28: A Multi-Component Herbal Preparation with Retinoid-Like Dermal Activity but Without Epidermal Effects Muhammad Nadeem Aslam, Helene Fligiel,

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PADMA 28: A Multi-Component Herbal Preparation with Retinoid-Like Dermal Activity but Without Epidermal Effects Muhammad Nadeem Aslam, Helene Fligiel, Humaira Lateef, Gary J. Fisher, Isaac Ginsburg, James Varani Journal of Investigative Dermatology Volume 124, Issue 3, Pages 524-529 (March 2005) DOI: 10.1111/j.0022-202X.2004.23596.x Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Effects of PADMA 28 on human dermal fibroblast proliferation in (A) keratinocyte basal medium (KBM) and (B) Ca2+-supplemented KBM. Proliferation was assessed as described in the Materials and Methods section. Values shown are means and standard deviations based on seven independent experiments with duplicate values per data point. Statistical significance of the differences between control and PADMA-treated groups was determined using ANOVA followed by paired-group comparisons. *p<0.05. Journal of Investigative Dermatology 2005 124, 524-529DOI: (10.1111/j.0022-202X.2004.23596.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Effects of PADMA 28 on matrix metalloproteinase (MMP)-1 and MMP-2 levels in fibroblast cell culture fluid from cells grown in keratinocyte basal medium (KBM) and Ca2+-supplemented KBM. MMP-1 and MMP-2 levels were assessed by β-casein and gelatin zymography, respectively, and quantified by scanning densitometry. Values shown are means and SD based on four independent experiments. Statistical significance of the differences between control and PADMA-treated groups was determined using Student's t test. *p<0.05. Inset: β-casein zymogram showing differences in MMP-1 levels between control and treated groups and gelatin zymogram showing no difference in MMP-2 levels between control and treated groups. Journal of Investigative Dermatology 2005 124, 524-529DOI: (10.1111/j.0022-202X.2004.23596.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Effects of PADMA 28 on matrix metalloproteinase (MMP)-1, MMP-2, and type I procollagen levels in human skin organ culture fluid. MMP levels were assessed by β-casein and gelatin zymography and quantified as described in the Materials and Methods section. Type I procollagen was assessed by ELISA as described in the Materials and Methods section. Values shown are means and SD based on four independent experiments. Statistical significance of the differences between control and PADMA-treated groups was determined using Student's t test. *p<0.05. Journal of Investigative Dermatology 2005 124, 524-529DOI: (10.1111/j.0022-202X.2004.23596.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Histological features of human skin after incubation for 8 d in organ culture in the absence or presence of PADMA 28. (A) Ca2+-supplemented KBM and (B) Ca2+-supplemented keratinocyte basal medium (KBM) plus 20 μg per mL PADMA 28. Normal histological features are observed under both conditions. (C) KBM and (D) KBM plus 20 μg per mL PADMA 28. There is extensive necrosis in the skin in the control section, but necrosis is less apparent in the skin treated with PADMA 28. Scale bar=24 μm. Journal of Investigative Dermatology 2005 124, 524-529DOI: (10.1111/j.0022-202X.2004.23596.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Histological features of Rhino mouse skin after topical treatment for 21 days (A) vehicle (dimethyl sulfoxide (DMSO)) alone, (B) PADMA 28 (500 mg per mL in DMSO), or (C) retinoic acid (RA) (10 mg per mL in DMSO). Mouse skin treated with vehicle alone or with PADMA 28 has normal histological features including a thin epidermis and a lack of inflammation. Epidermal hyperplasia along with perivascular accumulation of mononuclear cells and dermal microabscesses containing polymorphonuclear leukocytes is observed in RA-treated skin. Scale bar=57 μm. Journal of Investigative Dermatology 2005 124, 524-529DOI: (10.1111/j.0022-202X.2004.23596.x) Copyright © 2005 The Society for Investigative Dermatology, Inc Terms and Conditions