{"paper_id":"9cdf6bd1-5d71-403d-89c9-465d90e86341","body_text":"Abstract\nThe extracellular matrix (ECM) critically influences cell behavior, yet its properties in human endometrial lesions (HEL) and human uterine fibromas (HUF) are not well characterized. This study aimed to characterize their ECM and evaluate its impact on cell engraftment and proliferation while optimizing a decellularization protocol. HEL and HUF tissues, collected during laparoscopic surgeries, were decellularized using a novel protocol. Complete cell removal and preserved ECM microstructure were confirmed by histology, DAPI, Masson’s trichrome staining and scanning electron microscopy. The decellularized scaffolds were used as a platform for three-dimensional culture of human endometrial-derived mesenchymal stem cells (hEMSCs), with HUF serving as a fibrotic control originated from the same organ system. The biological impact of the ECM was assessed via immunohistochemistry for engraftment marker matrix metalloproteinase-9 and proliferation marker antigen Kiel-67. The in vivo recellularization potential of HEL scaffolds was further evaluated in a rat model, with HEL scaffold group at two timepoints (n = 6/group) and a sham control (n = 3). Results confirmed complete decellularization with maintained ECM integrity in both HEL and HUF. In vitro evaluation indicated that hEMSCs seeded more efficiently onto HEL scaffolds (51.16 ± 28.84) compared to HUF scaffolds (6.16 ± 7.29) (p = 0.012). The in vivo peritoneal implanted HEL scaffolds demonstrated significant time-dependent host cell recruitment and remodeling compared to the sham control. In conclusion, the decellularized HEL scaffold provides a superior ECM platform for cell seeding and engraftment compared to HUF, making it a promising platform for modeling endometriosis in both in vitro and in vivo settings.\nSimilar content being viewed by others\nData Availability\nThe datasets used and/or analyzed during the current study are available from the co-corresponding authors on reasonable request.\nAbbreviations\n- dHEL:\n-\nDecellularized human endometrial lesion\n- dHUF:\n-\nDecellularized human uterine fibroma\n- DAPI:\n-\n4’ ,6 -diamidino-2-phenylindole\n- DMSO:\n-\nDimethyl sulfoxide\n- ECM:\n-\nExtracellular matrix\n- EDTA:\n-\nEthylenediamine tetra-acetic acid\n- HEL:\n-\nHuman Endometrial Lesion\n- H&E:\n-\nHematoxylin & eosin staining\n- hEMSCs:\n-\nHuman endometrial derived mesenchymal stem cells\n- HUF:\n-\nHuman uterine fibroma\n- IHC:\n-\nImmunohistochemistry staining\n- Ki-67:\n-\nAntigen Kiel-67\n- MMP-9:\n-\nMatrix metalloproteinase-9\n- MTT:\n-\n3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide\n- OD:\n-\nOptical density\n- PBS:\n-\nPhosphate buffered sodium\n- SDS:\n-\nSodium dodecyl sulfate\n- SEM:\n-\nScanning electron microscopy\nReferences\nYoshimasa Y, Takao T, Katakura S, Tomisato S, Masuda H, Tanaka M, et al. 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Reproductive Sci. 2018;25(3):329–40.\nAcknowledgments\nWe express our profound gratitude to Dr. Masoomeh Shiravi Khoozani for her invaluable contribution in providing histopathological details of our study's in vitro and in vivo explants. Her insights and expertise were instrumental in enhancing the quality of our study, and we are deeply thankful for her dedicated involvement. Fig. 1 is “Created with BioRender.com”.\nFunding\nNo funding was received for conducting this study.\nAuthor information\nAuthors and Affiliations\nContributions\nAll authors contributed to the study conception and design. Conceptualization and Methodology were designed by Abdol-Mohammad Kajbafzadeh and Masoumeh Majidi Zolbin. Material preparation, data collection and analysis were performed by Ashkan Azimzadeh, Roxana Sahmani, Bahareh Mohammadi, Saman Behboodi Tanourlouee, Masoumeh Ekhtiari, Negar Mohammadi Ganjaroudi, Parmida Sadat Pezeshki, and Ali Mohebbi. The first draft of the manuscript was written by Negar Mohammadi Ganjaroudi and all authors commented on previous versions of the manuscript. All authors read and approved of the final manuscript.\nCorresponding authors\nEthics declarations\nEthics Approval\nAll procedures were performed in accordance with the Tehran University of Medical Sciences’ Guidelines and approved by the university’s Ethics Committee (IR.TUMS.VCR.REC.1398.406). The study was conducted in accordance with the principles outlined in the Declaration of Helsinki. Informed consent achieved from all premenopausal patient cases undergoing laparoscopy or laparotomy regarding treatment of endometriosis or uterine fibroma, describing the procedure and aims of the study in compliance with the regulations concerning the use of human tissues. All the animal procedures were approved by the animal ethics committee of the Tehran University of Medical Sciences, School of Medicine and Education Section of Basic Sciences and undergone surgery in accordance with the animal welfare Act and Guide for The Care and Use of Laboratory Animals and ARRIVE guidelines.\nCompeting Interests\nThe authors have no competing interests to declare relevant to this article’s content.\nAdditional information\nPublisher’s Note\nSpringer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.\nAshkan Azimzadeh and Roxana Sahmani are equally contributed as co-first authors.\nRights and permissions\nSpringer 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.\nAbout this article\nCite this article\nAzimzadeh, A., Sahmani, R., Mohammadi Ganjaroudi, N. et al. Human Endometriosis Scaffold Can Enhance Cell Seeding and Engraftment; an In Vitro and In Vivo Study. Reprod. Sci. 33, 84–93 (2026). https://doi.org/10.1007/s43032-025-02032-0\nReceived:\nAccepted:\nPublished:\nVersion of record:\nIssue date:\nDOI: https://doi.org/10.1007/s43032-025-02032-0","source_license":"public-domain-us","license_restricted":false}