Pentoxifylline Attenuates Transforming Growth Factor-β1-Stimulated Collagen Deposition and Elastogenesis in Human Tunica Albuginea-Derived Fibroblasts.

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Pentoxifylline Attenuates Transforming Growth Factor-β1-Stimulated Collagen Deposition and Elastogenesis in Human Tunica Albuginea-Derived Fibroblasts Part 1: Impact on Extracellular Matrix Alan W. Shindel, MD, Guiting Lin, MD, PhD, Hongxiu Ning, PhD, Lia Banie, BS, Yun-Ching Huang, MD, Gang Liu, MD, PhD, Ching- Shwun Lin, PhD, Tom F. Lue, MD The Journal of Sexual Medicine Volume 7, Issue 6, Pages 2077-2085 (June 2010) DOI: 10.1111/j.1743-6109.2010.01790.x Copyright © 2010 International Society for Sexual Medicine Terms and Conditions

Figure 1 Tunica albuginea-derived fibroblast cells were treated with vehicle or transforming growth factor-β1 (TGF-β1; 1ng/mL) in Dulbecco's modified Eagle medium with 5% fetal bovine serum for 14 days. In the native status (vehicle treated; A, B), there were no mature elastic fibers formed although small quantities of tropoelastin were identified (arrowheads). After treatment with TGF-β1, elastogenesis was activated. Many matured elastic fibers (arrows) were formed in the extracellular matrix (green) (C, D). Elastin fiber formation was more robust in PT cells. The Journal of Sexual Medicine 2010 7, 2077-2085DOI: (10.1111/j.1743-6109.2010.01790.x) Copyright © 2010 International Society for Sexual Medicine Terms and Conditions

Figure 2 Pentoxifylline (PTX) attenuates elastogenesis activated by transforming growth factor-β1 (TGF-β1) in tunica albuginea-derived fibroblast. PT TADF cells were pretreated with 0, 0.01, 10, and 100uM PTX for 6 hours followed by 1ng/mL TGF-β1 for 14 days in Dulbecco's modified Eagle medium with 5% fetal bovine serum. Inhibition of elastogenesis showed a dose–response increase with progressively higher levels of PTX in PT cells; a PTX dose–response relationship was not clearly evident in normal tunica cells (arrows indicate elastic fibers, arrowhead indicates tropoelastin. ×200). The Journal of Sexual Medicine 2010 7, 2077-2085DOI: (10.1111/j.1743-6109.2010.01790.x) Copyright © 2010 International Society for Sexual Medicine Terms and Conditions

Figure 3 Pentoxifylline (PTX) attenuates elastogenesis activated by transforming growth factor-β1 (TGF-β1) in normal tunica (NT) and PT cells. (A) PT tunica albuginea-derived fibroblast cells were treated with TGF-β1 (1ng/mL) or combination TGF-β1 (1ng/mL) and PTX (100uM). PTX attenuated elastogenesis in both PT and NT cells. The Journal of Sexual Medicine 2010 7, 2077-2085DOI: (10.1111/j.1743-6109.2010.01790.x) Copyright © 2010 International Society for Sexual Medicine Terms and Conditions

Figure 4 Pentoxifylline (PTX) decreases collagen I in tunica albuginea-derived fibroblast. Normal tunica (NT) and PT were treated with transforming growth factor-β1 (TGF-β1), TGF-β followed by PTX, and PTX followed by TGF-β1. Expression of collagen I was higher in PT relative to NT in the untreated control group (*P<0.05). TGF-β1 significantly increased collagen I in NT and PT cells from 0.169±0.02 and 0.703±0.1ug/mL to 1.342±0.01 and 1.8525±0.05ug/mL, respectively (#P<0.05 vs. controls). This effect was not observed in cells pretreated with PTX prior to TGF-β1 (0.569±0.02 and 0.75±0.05ug/mL in NT and PT, respectively, P<0.05 vs. TGF-β1 treated groups) although collagen I production in these NT cells remained significantly higher than what was observed in NT controls (#P<0.05 vs. control cells). PTX following TGF-β1 treatment did not affect collagen I increase relative to TGF-β1 alone (P>0.05; *P<0.05 vs. NT cells, #P<0.05 vs. control cells [PT and NT]). The Journal of Sexual Medicine 2010 7, 2077-2085DOI: (10.1111/j.1743-6109.2010.01790.x) Copyright © 2010 International Society for Sexual Medicine Terms and Conditions