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Mechanobiology in the Reproductive Tract
Published in Jiro Nagatomi, Eno Essien Ebong, Mechanobiology Handbook, 2018
Julie Anne MacDonald, Dori C. Woods
Previously discussed were several studies that defined the altered mechanosensitive state of fibroids in comparison to healthy myometrium tissue (see Section 22.5). Proposed new treatments seek to alter those changes and reinstate “healthy” mechanical signaling. One in vitro study found that treatment of three-dimensional leiomyoma cell cultures with a Rho-kinase inhibitor, fasudil, decreased the typically constituently high levels of active RhoA, as well as decreasing cellular contractility (Malik et al. 2014). Additional work from the same group also showed the inhibition of elevated integrin signaling in fibroid cells, through antibody-mediated inhibition of the β1 integrin subunit, also decreased Rho signaling (Malik et al. 2012). Reconstruction of the aberrant leiomyoma ECM is another strategy to restore tissue homeostasis. Jayes et al. propose to dissolve uterine fibroids in vivo through direct injection of collagenase Clostridium histolyticum, a proteolytic enzyme that specifically cleaves types I, II, and III collagens, typically overexpressed in fibroid masses (2016). Tissue isolated from patient fibroids was treated with collagenase C histolyticum and showed decreases in both rigidity and fibrosis following treatment (Jayes et al. 2016); however, this work has not yet moved to clinical trials.
Comparison of adipose stem cells sources from various locations of rat body for their application for seeding on polymer scaffolds
Published in Journal of Biomaterials Science, Polymer Edition, 2019
Agata Kurzyk, Tomasz Dębski, Wojciech Święszkowski, Zygmunt Pojda
Briefly, raw lipoaspirates were isolated as described previously [18]. Raw lipoaspirates were washed extensively with sterile phosphate-buffered saline (PBS) (Life Technologies, USA) to eliminate contaminating debris and red blood cells. Washed aspirates were treated with 0.075% collagenase (Clostridium histolyticum; type I; Sigma Aldrich, St. Louis, MO, USA) in PBS for 60 min at 37 °C with gentle agitation. Thereafter, collagenase was inactivated with 10% fetal bovine serum (FBS, Gibco®, USA) and the infranatant fluid was centrifuged (400 ×g for 10 min at 23 °C) until phase separation. To filter out larger tissue particles, the stromal vascular fraction (SVF) pellet was resuspended and passed through a 100-μm filter placed atop a new 50-ml centrifuge tube to facilitate gravitational separation. The filtrate was centrifuged at 400 ×g for 10 min to obtain a high-density SVF pellet containing ASCs, which was then cultured in tissue-culture treated culture dishes in medium comprising an equal volume of low-glucose Dulbecco’s modified Eagle’s medium (DMEM, Gibco®, USA) and fetal bovine serum (FBS) and incubated in a humidified atmosphere at 37 °C and 5% CO2. Media were replaced twice per week, and cells were passaged on approaching 80–90% confluence, using 0.25% trypsin-EDTA solution (Invitrogen, USA). Routine tests were performed before ASC preparation and after preparation, including sterility control (BACTEC, Bact/Alert, Becton Dickinson, blood agar, Columbia agar, and BHI agar), cell enumeration (Bürker chamber), and viability analysis (fluorescence microscopy, acridine orange plus ethidium bromide staining).
Devices for penile traction: the long and winding road to treating Peyronie’s disease
Published in Expert Review of Medical Devices, 2018
Shaan A Setia, Laurence A Levine
Most oral therapies (e.g. pentoxifylline, vitamin E, Potaba, colchicine) are thought to reduce inflammation and/or decrease fibrotic activity [29–33]. However, multiple studies have shown a lack of efficacy, particularly as monotherapy. A literature review from 2012 reports that there is no oral therapy that reliably reduces the signs and symptoms of PD in a clinically meaningful way [34]. ILI, on the other hand, represents an option more likely to provide meaningful benefit. Multiple different medications have been used with varying success. Calcium channel blockers such as verapamil or nicardipine have been shown to decrease fibroblast proliferation, increase collagenase activity, and decrease fibroblast ECM production in vitro [35]. Clinically, there are no large-scale, randomized placebo-controlled trials for calcium channel blockers. However, multiple small trials have shown some benefit [36–39]. Similar to calcium channel blockers, interferon injections have also demonstrated an ability to reduce fibroblast proliferation, increase collagenase activity, and decrease ECM production in vitro [40]. However, clinical outcomes have been more mixed [41–43]. The only US Food and Drug Administration approved nonsurgical treatment option for PD is collagenase Clostridium histolyticum (CCH). Its approval was largely based on the outcomes of phase 3 randomized, double-blind, placebo-controlled trials in 2013 (IMPRESS I and II) [44]. Collagenase is an enzyme which breaks down collagen. The IMPRESS trials demonstrated a statistically significant decrease in penile curvature, symptom bother, and low serious adverse events in men using CCH injections versus placebo.