Effect of hydrogel based on tissue extract of uterine leiomyoma enhances osteogenic activity of osteoblasts

Background. The extracellular matrix plays a key role in maintaining homeostasis and bone tissue regeneration by defining the conditions for cell differentiation and intercellular interactions. The native extracellular matrix is a complex three-dimensional network of structural proteins, glycoproteins, and proteoglycans that provide mechanical support to cells and transmit regulatory signals. 2D-culture systems do not reproduce the complex organization of the cellular microenvironment; therefore, the development of 3D-systems for bone tissue differentiation and remodeling in vitro remains an important challenge. Objective. To evaluate the biocompatibility of the hydrogel based on a tissue extract of uterine leiomyoma and its potential to regulate osteogenic differentiation and mineralization of osteoblasts in mono- and co-culture with endothelial cells. Materials and Methods. The tissue extract of uterine leiomyoma was obtained using a modified protocol originally developed for producing Matrigel®. Osteoblasts were cultured within the hydrogel in mono- and co-culture with endothelial cells under osteogenic conditions. Cell viability and morphology were assessed by live-cell microscopy. Mineralization of the extracellular matrix was evaluated using Alizarin Red S staining. Results. The hydrogel based on tissue extract of uterine leiomyoma demonstrated high biocompatibility toward osteoblasts, supporting their viability, promoting differentiation into osteocyte-like cells, and enhancing extracellular matrix mineralization. In co-culture with endothelial cells, these effects were further enhanced, indicating the important role of intercellular interactions. Conclusion. The hydrogel based on tissue extract of uterine leiomyoma represents a promising 3D culture model for the studying osteogenic differentiation of osteoblasts.

1. Andreeva ER, Matveeva DK, Zhidkova OV, Buravkova LB. Extracellular matrix as a factor regulating the physiological microenvironment of the cell. Progress in Physiological Science. 2024;55(1):16‒30. (In Russ.) https://doi.org/10.31857/s0301179824010033, https://elibrary.ru/xjcavk

2. Cheong S, Peng Y, Lu F, He Y. Structural extracellular matrix-mediated molecular signaling in wound repair and tissue regeneration. Biochimie. 2025;229:58–68. https://doi.org/10.1016/j.biochi.2024.10.003

3. Kamal KM, Ghazali AR, Selvarajah GT, et al. The extracellular matrix: structure, composition, biological functions, diseases, and therapeutic targets. Molecular Biomedicine. 2026;7(1):38. https://doi.org/10.1186/s43556-026-00436-1

4. Liu N, Shi Y, Li J, et al. Morphology-guided cellular behavior modulation with 3D-printed engineered ECM. Cell Biomaterials. 2025;1:100090. https://doi.org/10.1016/j.celbio.2025.100090

5. Zhao T, Huang Y, Zhu J, et al. Extracellular matrix signaling cues: biological functions, diseases, and therapeutic targets. MedComm (Beijing). 2025;6. https://doi.org/10.1002/mco2.70281

6. Kolb AD, Bussard KM. The Bone extracellular matrix as an ideal milieu for cancer cell metastases. Cancers. 2019;11(7):1020. https://doi.org/10.3390/cancers11071020

7. Lin X, Patil S, Gao Y-G, Qian A. The bone extracellular matrix in bone formation and regeneration. Front Pharmacol. 2020;11. https://doi.org/10.3389/fphar.2020.00757

8. Cai W, Huo Y, Liu Y, et al. Biomechanics in bone regeneration and mechanobiology in osteoblasts: Fundamental concepts and recent progress. Eng Medicine. 2025;2:100057. https://doi.org/10.1016/j.engmed.2025.100057

9. Li M, Zhang A, Li J, et al. Osteoblast/fibroblast coculture derived bioactive ECM with unique matrisome profile facilitates bone regeneration. Bioact Mater. 2020;5(4):938–48. https://doi.org/10.1016/j.bioactmat.2020.06.017

10. Perepletchikova D, Malashicheva A. Communication between endothelial cells and osteoblasts in regulation of bone homeostasis: Notch players. Stem Cell Res Ther. 2025;16(1):56. https://doi.org/10.1186/s13287-025-04176-x

11. Dalle Carbonare L, Cominacini M, Trabetti E, et al. The bone microenvironment: new insights into the role of stem cells and cell communication in bone regeneration. Stem Cell Res Ther. 2025;16(1):169. https://doi.org/10.1186/s13287-025-04288-4

12. Huang G, Hou T, Song D, Meng T. The regulatory networks and mechanisms of bone microenvironment in tumorigenesis and metastasis. J Bone Oncol. 2025;55:100729. https://doi.org/10.1016/j.jbo.2025.100729

13. Amirazad H, Dadashpour M, Zarghami N. Application of decellularized bone matrix as a bioscaffold in bone tissue engineering. J Biol Eng. 2022;16:1. https://doi.org/10.1186/s13036-021-00282-5

14. Li C, An N, Song Q, et al. Enhancing organoid culture: harnessing the potential of decellularized extracellular matrix hydrogels for mimicking microenvironments. J Biomed Sci. 2024;31(1):96. https://doi.org/10.1186/s12929-024-01086-7

15. Guo X, Liu B, Zhang Y, et al. Decellularized extracellular matrix for organoid and engineered organ culture. J Tissue Eng. 2024;15. https://doi.org/10.1177/20417314241300386

16. Kibbey MC. Maintenance of the EHS sarcoma and Matrigel preparation. Journal of Tissue Culture Methods. 1994;16(3–4):227–230. https://doi.org/10.1007/BF01540656

17. Wolff L, Hendrix S. Rethinking Matrigel: The complex journey to matrix alternatives in organoid culture. Advanced Science. 2025;12:47. https://doi.org/10.1002/advs.202508734

18. Islam MS, Ciavattini A, Petraglia F, et al. Extracellular matrix in uterine leiomyoma pathogenesis: a potential target for future therapeutics. Hum Reprod Update. 2018;24(1):59–85. https://doi.org/10.1093/humupd/dmx032

19. Saad EE, Michel R, Borahay MA. Immunosuppressive tumor microenvironment and uterine fibroids: Role in collagen synthesis. Cytokine Growth Factor Rev. 2024;75:93–100. https://doi.org/10.1016/j.cytogfr.2023.10.002

20. Yang Q, Ciebiera M, Bariani MV, et al. Comprehensive review of uterine fibroids: developmental origin, pathogenesis, and treatment. Endocr Rev. 2022;43(4):678–719. https://doi.org/10.1210/endrev/bnab039

21. Kostina D, Lobov A, Klausen P, et al. Isolation of human osteoblast cells capable for mineralization and synthetizing bone-related proteins in vitro from adult bone. Cells. 2022;11(21):3356. https://doi.org/10.3390/cells11213356

22. Kottmann V, Nienhaus M, et al. From bone homeostasis to skeletal metastasis and osteosarcoma: Insights into osteoclast and osteoblast roles in bone remodelling and cancer. Biochimica et Biophysica Acta ‒ Reviews on Cancer. 2026;1881(2):189551. https://doi.org/10.1016/j.bbcan.2026.189551

23. Li S, Cai X, Guo J, et al. Cell communication and relevant signaling pathways in osteogenesis–angiogenesis coupling. Bone Res. 2025;13(1):45. https://doi.org/10.1038/s41413-025-00417-0

24. Ma C, Du T, Niu X, Fan Y. Biomechanics and mechanobiology of the bone matrix. Bone Res. 2022;10(1):59. https:// doi.org/10.1038/s41413-022-00223-y

25. Emami A, Izadi E, Oskouie IM. Preservation of extracellular matrix in decellularized bone scaffolds: Strategies, challenges, and future directions. Tissue Cell. 2025;97:103047. https://doi.org/10.1016/j.tice.2025.103047

26. Li M, Liu X, Tian E, Liao W. The role of collagen in mechanotransduction and its influence on bone metabolic activity. Int J Biol Macromol. 2025;318:144968. https://doi.org/10.1016/j.ijbiomac.2025.144968

27. Brown M, Li J, Moraes C, et al. Decellularized extracellular matrix: New promising and challenging biomaterials for regenerative medicine. Biomaterials. 2022;289:121786. https:// doi.org/10.1016/j.biomaterials.2022.121786

28. Guo W-Y, Wang W-H, et al. Decellularised extracellular matrix-based injectable hydrogels for tissue engineering applications. Biomaterials translational. 2024;5(2):114–128. https://doi.org/10.12336/biomatertransl.2024.02.003

29. Gkantzou E, Rodríguez-Rojas A, et al. Decellularized extracellular matrix for organoids development and 3D Bioprinting. Organoids. 2026;5(1):2. https://doi.org/10.3390/organoids5010002

30. Zhang X, Chen X, et al. Decellularized extracellular matrix scaffolds: Recent trends and emerging strategies in tissue engineering. Bioact Mater. 2022;10:15–31. https://doi.org/10.1016/j.bioactmat.2021.09.014

31. Yang H, Xia J, et al. From production to the clinic: decellularized extracellular matrix as a biomaterial for tissue engineering and regenerative medicine. Bioengineering. 2025;13(1):24. https://doi.org/10.3390/bioengineering13010024

32. Liang R, Pan R, et al. Decellularized extracellular matrices for skin wound treatment. Materials. 2025;18(12):2752. https://doi.org/10.3390/ma18122752

33. Zhu Y, Yang W, et al. Constructing biomimetic microenvironments for liver regeneration. J Nanobiotechnology. 2025;23(1):655. https://doi.org/10.1186/s12951-025-03729-9

34. Kim JW, Nam SA, et al. Kidney decellularized extracellular matrix enhanced the vascularization and maturation of human kidney organoids. Advanced Science. 2022;9(15). https://doi.org/10.1002/advs.202103526

35. Liu W, Zhang X, et al. Decellularized extracellular matrix materials for treatment of ischemic cardiomyopathy. Bioact Mater. 2024;33:460–482. https://doi.org/10.1016/j.bioactmat.2023.10.015

36. Rusinova TV, Vinogradov RA, Asyakina AS, et al. Decellularized nerve scaffold in a rat model of extensive peripheral nerve damage. Modern Technologies in Medicine. 2025;17(6):16. https://doi.org/10.17691/stm2025.17.6.02

37. Kim Y-H, Cidonio G, Kanczler JM, et al. Human bone tissue-derived ECM hydrogels: Controlling physicochemical, biochemical, and biological properties through processing parameters. Bioact Mater. 2025;43:114–128. https://doi.org/10.1016/j.bioactmat.2024.09.007

38. Go YY, Kim SE, Cho GJ, et al. Differential effects of amnion and chorion membrane extracts on osteoblast-like cells due to the different growth factor composition of the extracts. PLoS One. 2017;12(8):e0182716. https://doi.org/10.1371/journal.pone.0182716

39. Kang B-J, Ryu H-H, Park S-S, et al. Effect of Matrigel on the osteogenic potential of canine adipose tissue-derived mesenchymal stem cells. J Vet Med Sci. 2012;74(7):827–836. https://doi.org/10.1292/jvms.11-0484

40. Tangporncharoen R, Silathapanasakul A, Tragoonlugkana P, et al. The extracts of osteoblast developed from adipose-derived stem cell and its role in osteogenesis. J Orthop Surg Res 2024;19(1):255. https://doi.org/10.1186/s13018-024-04747-3

41. Wei B, Chen Y, Zhang S, et al. Proteins extracted from placenta regulate osteogenic differentiation of human mesenchymal stem cells. Tissue Cell. 2025;96:103015. https:// doi.org/10.1016/j.tice.2025.103015

42. Coyle A, Chakraborty A, et al. In vitro engineered ECM-incorporated hydrogels for osteochondral tissue repair: a cell-free approach. Adv Healthc Mater. 2025;14(4). https://doi.org/10.1002/adhm.202402701

43. Dekker M, Hipwood L, et al. Bone-derived dECM hydrogels support tunable microenvironments for in vitro osteogenic differentiation. Adv Healthc Mater. 2026;15(1). https://doi.org/10.1002/adhm.202501350

44. Fu T, Liang P, et al. Matrigel scaffolding enhances BMP9-induced bone formation in dental follicle stem/precursor cells. Int J Med Sci. 2019;16(4):567–575. https://doi.org/10.7150/ijms.30801

45. Jeon J, Lee MS, Yang HS. Differentiated osteoblasts derived decellularized extracellular matrix to promote osteogenic differentiation. Biomater Res. 2018;22(1). https://doi.org/10.1186/s40824-018-0115-0

46. Nasello G, Alamán-Díez P, et al. Primary human osteoblasts cultured in a 3D microenvironment create a unique representative model of their differentiation into osteocytes. Front Bioeng Biotechnol. 2020;8. https://doi.org/10.3389/fbioe.2020.00336

47. Maggio N, Banfi A. The osteo-angiogenic signaling crosstalk for bone regeneration: harmony out of complexity. Curr. Opin. Biotechnol. 2022;76:102750. https://doi.org/10.1016/j.copbio.2022.102750

48. Perepletchikova D, Kuchur P, Basovich L, et al. Endothelial- mesenchymal crosstalk drives osteogenic differentiation of human osteoblasts through Notch signaling. Cell Communication and Signaling. 2025;23(1):100. https://doi.org/10.1186/s12964-025-02096-0

49. Wang T, Yao H, Jin F, et al. Bidirectional regulation between bone and vasculature: Mechanisms of osteogenesis and angiogenesis. J Adv Res. 2026. https://doi.org/10.1016/j.jare.2026.02.042

50. Yang Q, Liu S, et al. A new paradigm in bone tissue biomaterials: Enhanced osteogenesis–angiogenic coupling by targeting H-type blood vessels. Biomaterials. 2026;324:123423. https://doi.org/10.1016/j.biomaterials.2025.123423

51. Laude M, Kolliopoulos V, et al. Extracellular-matrix-based materials from decellularized tissue: opportunities, challenges, and future directions in regenerative medicine. Adv Healthc Mater. 2026;15(1). https://doi.org/10.1002/adhm.202502107

52. Parasuraman G, Rani J MS, et al. Matrigel-encapsulated articular cartilage derived fibronectin adhesion assay derived chondroprogenitors for enhanced chondrogenic differentiation: An in vitro evaluation. Tissue Cell. 2025;92:102638. https://doi.org/10.1016/j.tice.2024.102638