Assessing the Impact of Collagenase Clostridium histolyticum Injection on Biomechanical Properties of Uterine Fibroids and Adjacent Myometrial Tissue

Uterine fibroids (UF) are common benign tumors characterized by a higher collagen content compared to normal myometrium. Existing treatment options provide temporary relief or may impact fertility, underscoring the need for a fertility-preserving, non-invasive treatment alternative. This study aimed to explore the impact of collagenase Clostridium histolyticum (CCH) injection on the biomechanical properties of uterine fibroids to assess its potential as a therapeutic intervention. Between January 2018 and November 2020, 17 uterine fibroids measuring 30-50 mm in length with adjacent myometrium of 30 mm were obtained from patients undergoing hysterectomy for abnormal uterine bleeding. Each sample was divided into two halves, with one serving as a control and the other as an intervention. The control group samples were processed within 24 hours of collection, while the intervention group samples were injected with CCH at a dose normalized to tissue volume (2 mg/cm³) and incubated for 24 hours prior to mechanical testing. Tissue pieces measuring 30 mm in length, 12 mm in width, and 2-3 mm in thickness were prepared for mechanical testing. A tensile test was conducted using a tensile machine at a fixed speed rate of 10 mm/min under ambient temperature conditions. The strain measured displacement, UTS indicated the stiffness of the samples. The results were presented graphically on a stress-strain curve. In fibroid tissues, UTS decreased significantly from 1.79 ± 0.07 MPa in controls to 0.40 ± 0.04 MPa in treated samples (Δ = −1.39, P < 0.0001). Young’s Modulus reduced from 0.343 ± 0.068 MPa to 0.069 ± 0.031 MPa (Δ = −0.274, P = 0.00332). Conversely, myometrium tissues exhibited a modest reduction in UTS from 1.77 ± 0.04 MPa to 1.45 ± 0.05 MPa (Δ = −0.32, P < 0.0001), with no significant change in Young’s Modulus (0.164 ± 0.129 MPa to 0.108 ± 0.104 MPa, Δ = −0.056, P = 0.94). Elongation at rupture increased in both tissues: fibroids from 4.85 ± 0.15% to 4.96 ± 0.10% (Δ = +0.11, P = 0.78), and myometrium from 7.12 ± 0.06% to 9.01 ± 0.05% (Δ = +1.89, P < 0.0001). Collagenase treatment significantly weakened fibroids by reducing tensile strength and stiffness, while myometrium showed reduced strength but increased elongation due to apoptosis and reduced cell contractility. The results demonstrate that intrafibroid collagenase injections selectively soften fibroids without compromising the myometrium, highlighting its potential as a minimally invasive fibroid treatment option. Similar content being viewed by others Change history 26 February 2026 A Correction to this paper has been published: https://doi.org/10.1007/s43032-026-02071-1

Hervé F, Katty A, Isabelle Q, Céline S. Impact of uterine fibroids on quality of life: a national cross-sectional survey. Eur J Obstet Gynecol Reprod Biol. 2018;229:32–7. Ahmad A, Kumar M, Bhoi NR, Badruddeen AJ, Khan MI, et al. Diagnosis and management of uterine fibroids: current trends and future strategies. J Basic Clin Physiol Pharmacol. 2023;34(3):291–310. Stewart EA, Laughlin-Tommaso SK, Catherino WH, Lalitkumar S, Gupta D, Vollenhoven B. Uterine fibroids. Nat Rev Dis Primers. 2016;2:16043. Ciebiera M, Ali M, Prince L, Jackson-Bey T, Atabiekov I, Zgliczyński S, et al. The evolving role of natural compounds in the medical treatment of uterine fibroids. J Clin Med. 2020. https://doi.org/10.3390/jcm9051479. Najafiarab H, Farzaneh F, Talebian N, Ghasemi Z, Talayeh M. Health-related quality of life among Iranian women with uterine fibroids: a cross sectional study. J Family Reprod Health. 2024;18(3):164. Islam MS, Afrin S, Singh B, Jayes FL, Brennan JT, Borahay MA, et al. Extracellular matrix and hippo signaling as therapeutic targets of antifibrotic compounds for uterine fibroids. Clin Transl Med. 2021;11(7):e475. Mas A, Tarazona M, Dasí Carrasco J, Estaca G, Cristóbal I, Monleón J. Updated approaches for management of uterine fibroids. Int J Womens Health. 2017;9:607–17. Saridogan E, Cutner A. Endoscopic management of uterine fibroids. Hum Fertil. 2006;9(4):201–8. Vilos GA, Allaire C, Laberge P-Y, Leyland N, Vilos AG, Murji A, et al. The management of uterine leiomyomas. J Obstet Gynaecol Can. 2015;37(2):157–78. Corder RD, Gadi SV, Vachieri RB, Jayes FL, Cullen JM, Khan SA, et al. Using rheology to quantify the effects of localized collagenase treatments on uterine fibroid digestion. Acta Biomater. 2021;134:443–52. Jayes FL, Liu B, Feng L, Aviles-Espinoza N, Leikin S, Leppert PC. Evidence of biomechanical and collagen heterogeneity in uterine fibroids. PLoS One. 2019;14(4):e0215646. Leppert PC, Baginski T, Prupas C, Catherino WH, Pletcher S, Segars JH. Comparative ultrastructure of collagen fibrils in uterine leiomyomas and normal myometrium. Fertil Steril. 2004;82:1182–7. Rogers R, Norian J, Malik M, Christman G, Abu-Asab M, Chen F, et al. Mechanical homeostasis is altered in uterine leiomyoma. Am J Obstet Gynecol. 2008;198(4):474.e1–11. Norian JM, Owen CM, Taboas J, Korecki C, Tuan R, Malik M, et al. Characterization of tissue biomechanics and mechanical signaling in uterine leiomyoma. Matrix Biol. 2012;31(1):57–65. Naomi R, Ridzuan PM, Bahari H. Current insights into collagen type I. Polymers. 2021. https://doi.org/10.3390/polym13162642. Brunengraber LN, Jayes FL, Leppert PC. Injectable Clostridium histolyticum collagenase as a potential treatment for uterine fibroids. Reprod Sci. 2014;21(12):1452–9. Toyoshima T, Matsushita O, Minami J, Nishi N, Okabe A, Itano T. Collagen-binding domain of a Clostridium histolyticum collagenase exhibits a broad substrate spectrum both in vitro and in vivo. Connect Tissue Res. 2001;42(4):281–90. Mallya SK, Mookhtiar KA, Van Wart HE. Kinetics of hydrolysis of type I, II, and III collagens by the class I and II Clostridium histolyticum collagenases. J Protein Chem. 1992;11(1):99–107. Singh B, Sims H, Trueheart I, Simpson K, Wang KC, Patzkowsky K, et al. A phase I clinical trial to assess safety and tolerability of injectable collagenase in women with symptomatic uterine fibroids. Reprod Sci. 2021;28(9):2699–709. Dhillon S. Collagenase Clostridium histolyticum in Peyronie’s disease: a guide to its use in the EU. Drugs Ther Perspect. 2016;32(8):311–6. Gelbard M, Goldstein I, Hellstrom WJ, McMahon CG, Smith T, Tursi J, et al. Clinical efficacy, safety and tolerability of collagenase clostridium histolyticum for the treatment of peyronie disease in 2 large double-blind, randomized, placebo controlled phase 3 studies. J Urol. 2013;190(1):199–207. Badalamente MA, Hurst LC. Efficacy and safety of injectable mixed collagenase subtypes in the treatment of Dupuytren's contracture. J Hand Surg Am. 2007;32(6):767–74. Thomas A, Bayat A. The emerging role of Clostridium histolyticum collagenase in the treatment of Dupuytren disease. Ther Clin Risk Manag. 2010;6:557–72. Han S, Makareeva E, Kuznetsova NV, DeRidder AM, Sutter MB, Losert W, et al. Molecular mechanism of type I collagen homotrimer resistance to mammalian collagenases. J Biol Chem. 2010;285(29):22276–81. Hurst LC, Badalamente MA, Hentz VR, Hotchkiss RN, Kaplan FTD, Meals RA, et al. Injectable collagenase clostridium histolyticum for Dupuytren’s contracture. N Engl J Med. 2009;361(10):968–79. Poullis C, Shabbir M, Eardley I, Mulhall J, Minhas S. Clostridium histolyticum collagenase - is this a revolutionary medical treatment for Peyronie’s disease? BJU Int. 2016;118(2):186–9. Mazza E, Parra-Saavedra M, Bajka M, Gratacos E, Nicolaides K, Deprest J. In vivo assessment of the biomechanical properties of the uterine cervix in pregnancy. Prenat Diagn. 2014;34(1):33–41. Jayes FL, Liu B, Moutos FT, Kuchibhatla M, Guilak F, Leppert PC, et al. Loss of stiffness in collagen-rich uterine fibroids after digestion with purified collagenase Clostridium histolyticum. Am J Obstet Gynecol. 2016;215(5):596.e1–8. Della Rocca Y, Fonticoli L, Rajan TS, Trubiani O, Caputi S, Diomede F, et al. Hypoxia: molecular pathophysiological mechanisms in human diseases. J Physiol Biochem. 2022;78(4):739–52. Meiss RA. Mechanics of smooth muscle contraction. Cellular aspects of smooth muscle function 1997:169–201. Funding This study was funded by the Iran University of Medical Sciences, Tehran, Iran. Author information Authors and Affiliations Contributions K.T conceptualized the idea, while K.T and R.Sh provided the specimen. A.Kh, A.F-A, L.A, K.T, A.K, and N.A performed the intervention process and conducted the tensile test. L.A conducted the formal analysis, while A.F-A handled the visualization. Sh.Ch and A.M supervised the experiment and provided funding. S.H, I.A, AFA and K.T reviewed the validity of the results. L.A and A.K and A.Kh drafted the initial manuscript, which was then reviewed by K.T, AFA, I.A, and S.H. Ethics declarations Ethical Considerations All participants were provided and signed an informed consent. The Ethics Committee of Iran University of Medical Sciences, Tehran, Iran, approved the study (ethics approval code: IR.IUMS.REC.1397.199). Conflict of interests The authors declare that they have no conflicts of interest. Additional information Publisher’s Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Tahermanesh, K., Karimzadeh, A., Allahqoli, L. et al. Assessing the Impact of Collagenase Clostridium histolyticum Injection on Biomechanical Properties of Uterine Fibroids and Adjacent Myometrial Tissue. Reprod. Sci. 33, 391–398 (2026). https://doi.org/10.1007/s43032-025-02035-x Received: Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s43032-025-02035-x