1. Lipodermatosclerosis is Characterized by Elevated Expression and Activation of Matrix Metalloproteinases: Implications for Venous Ulcer Formation 
Yared Herouy, Gudula Pornschlegel, Christoph Stetter, Harald Grenz, Erwin Schöpf, Johannes Norgauer, Wolfgang Vanscheidt 
Journal of Investigative Dermatology 
Volume 111, Issue 5, Pages (November 1998)
DOI: /j x
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

2. Figure 1
The mRNA expression of matrix metalloproteinases and their tissue inhibitors in skin lesions of lipodermatosclerosis. (A) Total mRNA was prepared from biopsy specimen of liposclerotic (left lanes) and healthy skin (right lanes). Reverse transcription and amplification of generated cDNA with polymerase chain reaction was performed. Amplified products of MMP-1, MMP-2, MMP-9, TIMP-1, TIMP-2, and the control gene GAPDH were stained by ethidium bromide. Representative data from one patient with lipodermatosclerosis and one control are shown. (B) Densitometric evaluation of generated products of MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 from liposclerotic lesions (black bars) and healthy controls (white bars). To compare mRNA expression of different patients, the obtained signals were normalized to the level of the control gene GAPDH by calculating the ratio between the analyzed genes and GAPDH. Data are means ± SEM (n = 6 liposclerotic lesions; n = 6 controls). The significance of difference was determined by an unpaired Student’s t test and is indicated in each mapped group [*p < 0.05, **p < 0.01, ***p < and not significant (n.s.)].
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

3. Figure 2
Protein expression of matrix metalloproteinases and their tissue inhibitors in extracts of liposclerotic skin lesions. (A, B) Western blot with monoclonal antibodies against MMP-1, MMP-2, MMP-9, TIMP-1, and TIMP-2 in liposclerotic skin lesions. A representative immunoblotting from one patient with lipodermatosclerosis and one control are shown. (C, D) Densitometric evaluations of immunoreactive bands for the active and the zymogen form of MMP-1, unbound TIMP-1, and the proMMP-1/TIMP-1 complex as well as for the active and the zymogen form of MMP-2, unbound TIMP-2, and the inactive proMMP-2/TIMP-2 complex from liposclerotic skin (black bars) and healthy skin (white bars). Data are means ± SEM (n = 7 for liposclerotic lesions; n = 6 for controls). The significance of difference was determined by an unpaired Student’s t test and is indicated in each mapped group [*p < 0.05, **p < 0.01, ***p < and not significant (n.s.)].
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

4. Figure 3
Identification of gelatinolytic proteins in extracts of liposclerotic lesions. (A) SDS-PAGE zymography of extracts from liposclerotic lesions (left lane) and healthy skin (right lane). Equal amounts of total protein were loaded on each lane of a gelatin-containing gel. All tissue extracts displayed gelatinolytic activity comigrating with the zymogen form of MMP-2, proMMP-2, MMP-9, and proMMP-9. Representative data from one patient with lipodermatosclerosis and one control are shown. The experiment was repeated with comparable results. (B) Densitometric evaluation of the gelatinolytic activity of tissue extracts of liposclerotic lesions (back bars) and from healthy skin (white bars). Data are means ± SEM (n = 5 for liposclerotic lesions; n =5 for controls). The significance of difference was determined by an unpaired Student’s t test and is indicated in each mapped group [*p < 0.05, **p < 0.01, ***p < and not significant (n.s.)].
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

5. Figure 4
Collagen degradation of skin lesions of lipodermatosclerosis. The proteolytic activity of tissue extracts from liposclerotic lesions (black bars) and healthy skin (white bars) was analyzed with [3H]labeled type I collagen and [3H]labeled type IV collagen. Data are means ± SEM (n = 5 for liposclerotic lesions; n = 5 for controls). The significance of difference was determined by an unpaired student’s t test and is indicated in each mapped group (*p < 0.05, **p < 0.01, ***p < 0.001).
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

6. Figure 5
Immunohistochemical detection of MMP-1, MMP-2, and TIMP-2 in lesions of lipodermatosclerosis. In lipodermatosclerosis MMP-1 displayed increased diffuse immunoreactivity in the epidermis and dermis (A). In contrast MMP-2 was localized in perivascular regions (B), deep reticular dermis (C), and the basal and suprabasal layer of the epidermis (D), whereas in healthy skin diffuse and weak staining pattern was detected. In healthy skin a continuous line of immunoreactivity against TIMP-2 along the basement membrane was found (E), whereas in liposclerotic lesions the signal was weak. Representative photographs from one patient and one control are shown. The experiment was repeated with comparable results (n = 6 for liposclerotic lesions; n = 6 for controls). Scale bars: (A, E) 500 μm; (B) 100 μm; (C) 20 μm; (D) 200 μm.
Journal of Investigative Dermatology  , DOI: ( /j x)
Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions