Role of matrix metalloproteinases 1, 2, and 9 and tissue inhibitor of matrix metalloproteinase-1 in chronic venous insufficiency Satoshi Saito, MD, Matthew.

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Role of matrix metalloproteinases 1, 2, and 9 and tissue inhibitor of matrix metalloproteinase-1 in chronic venous insufficiency Satoshi Saito, MD, Matthew J. Trovato, BA, Ruoxu You, MD, PhD, Brajesh K. Lal, MD, Funmi Fasehun, PhD, Frank T. Padberg, MD, Robert W. Hobson, MD, Walter N. Durán, PhD, Peter J. Pappas, MD Journal of Vascular Surgery Volume 34, Issue 5, Pages 930-938 (November 2001) DOI: 10.1067/mva.2001.119503 Copyright © 2001 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 1 Bar graph demonstrating results of quantitative reverse transcriptase PCR for MMP-1 and TIMP-1. mRNA levels for MMP-1 were increased in class-4 and class-6 patients (P ≤.01). Gene transcripts for TIMP-1 were increased in class-6 patients only (P ≤.05); however, all TIMP-1 transcripts were increased five-fold compared with MMP-1. Journal of Vascular Surgery 2001 34, 930-938DOI: (10.1067/mva.2001.119503) Copyright © 2001 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 2 Western blot demonstrating the latent band for MMP-9 from a lower-calf (LC ) biopsy in a patient with an active venous ulcer. The lower-calf and lower-thigh (LT ) biopsies did not demonstrate a band for the active component of MMP-9 (arrow indicates 90-kd band, corresponding to the latent form of MMP-9). Journal of Vascular Surgery 2001 34, 930-938DOI: (10.1067/mva.2001.119503) Copyright © 2001 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 3 Bar graph demonstrating tissue amounts of the active components of MMP-1, MMP-2, and TIMP-1. The active component of MMP-2 was increased in class-4 and class-5 patients relative to MMP-1 and TIMP-1 (P ≤.01). The active component of MMP-9 was not observed. Journal of Vascular Surgery 2001 34, 930-938DOI: (10.1067/mva.2001.119503) Copyright © 2001 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 4 Three-percent gel zymogram demonstrating gelatinolytic activity of CVI skin protein extracts. The 92-kd band represents the latent form of MMP-9. The 66-kd and 64-kd bands represent active forms of MMP-2. MWkDa, Molecular weight in kilodaltons. Journal of Vascular Surgery 2001 34, 930-938DOI: (10.1067/mva.2001.119503) Copyright © 2001 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 5 A, Dermal CVI biopsy of skin stained with hematoxylin alone (185×). Vertical arrow indicates basement membrane of epidermis. Horizontal arrow indicates dermal capillary. B , Dermal CVI skin biopsy. Dark brown granules indicate cells that are positive for MMP-1. Vertical arrows point to spindle-shaped cells that appear similar to fibroblasts (185×). The horizontal arrow indicates a MMP-1–positive leukocyte outside the perivascular cuff. C , Magnified view of right lower quadrant of Fig 5, B, demonstrating perivascular cuffs. Arrows indicate circular cells traversing the cuffs which appear morphologically similar to leukocytes (460×). D , Dermal CVI skin biopsy stained for MMP-2 and counterstained with hematoxylin. Blue dots are nuclei of dermal cells. Dark brown staining of the dermis is positive for MMP-2 (185×). E , Magnified view of right upper quadrant of Fig 5, D. Horizontal arrows indicate fibroblasts, and vertical arrows indicate leukocytes traversing perivascular cuffs (460×). F , Dermal CVI skin biopsy. Vertical arrows indicate TIMP-1– positive keratinocytes in the basal layer of the epidermis (185×). Journal of Vascular Surgery 2001 34, 930-938DOI: (10.1067/mva.2001.119503) Copyright © 2001 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions

Fig. 6 Diagram indicating the possible role of TGF-β1 and MMPs in venous ulcer formation. Journal of Vascular Surgery 2001 34, 930-938DOI: (10.1067/mva.2001.119503) Copyright © 2001 Society for Vascular Surgery and The American Association for Vascular Surgery Terms and Conditions