Paracrine activity of Smurf1-silenced mesenchymal stem cells enhances bone regeneration and reduces bone loss in postmenopausal osteoporosis.

Publisher: BioMed Central Country of Publication: England NLM ID: 101527581 Publication Model: Electronic Cited Medium: Internet ISSN: 1757-6512 (Electronic) Linking ISSN: 17576512 NLM ISO Abbreviation: Stem Cell Res Ther Subsets: MEDLINE

Original Publication: London : BioMed Central

Mesenchymal Stem Cells*/metabolism ; Ubiquitin-Protein Ligases*/metabolism ; Ubiquitin-Protein Ligases*/genetics ; Osteoporosis, Postmenopausal*/therapy ; Osteoporosis, Postmenopausal*/metabolism ; Bone Regeneration* ; Osteogenesis*; Animals ; Humans ; Female ; Rats ; Mice ; Paracrine Communication ; Culture Media, Conditioned/pharmacology ; Rats, Sprague-Dawley ; Disease Models, Animal

Background: Osteoporosis (OP), characterized by reduced bone mass and mineral density, is a global metabolic disorder that severely impacts the quality of life in affected individuals. Although current pharmacological treatments are effective, their long-term use is often associated with adverse effects, highlighting the need for safer, more sustainable therapeutic strategies. This study investigates the pro-osteogenic and anti-resorptive potential of the secretome from Smurf1-silenced mesenchymal stem cells (MSCs) as a promising cell-free therapy for bone regeneration.
Methods: Conditioned media (CM) from Smurf1-silenced rat (rCM-Smur1) and human MSCs (hCM-Smurf1) was collected and analyzed. Pro-osteogenic potential was assessed by measuring in vitro mineralization in human and rat MSCs cultures. In vivo, studies were conducted using a rat ectopic bone formation model and a post-menopausal osteoporotic mouse model. Additionally, primary human osteoporotic MSCs were preconditioned with hCM-Smurf1, and their osteogenic capacity was compared to that induced by BMP2 treatment. Ex vivo, human bone explants were treated with hCM-Smurf1 to assess anti-resorptive effects. Proteomic analysis of the soluble and vesicular CM fractions identified key proteins involved in bone regeneration.
Results: CM from Smurf1-silenced MSCs significantly enhanced mineralization in vitro and bone formation in vivo. Preconditioning human osteoporotic MSCs with hCM-Smurf1 significantly increases in vitro mineralization, with levels comparable to those achieved with BMP2 treatment. Additionally, in ex vivo human bone cultures, treatment with hCM-Smurf1 significantly reduced RANKL expression without affecting OPG levels, indicating an anti-resorptive effect. In vivo, CM from Smurf1-silenced MSCs significantly increased bone formation in a rat ectopic model, and its local administration reduced trabecular bone loss by 50% in a post-menopausal osteoporotic mouse model after a single administration within just four weeks. Proteomic analysis revealed both soluble and vesicular fractions of hCM-Smurf1 were enriched with proteins essential for ossification and extracellular matrix organization, enhancing osteogenic differentiation.
Conclusions: The Smurf1-silenced MSCs' secretome shows potent osteogenic and anti-resorptive effects, significantly enhancing bone formation and reducing bone loss. This study provides compelling evidence for the therapeutic potential of Smurf1-silenced MSC-derived secretome as a non-toxic and targeted treatment for osteoporosis. These findings warrant further in vivo studies and clinical trials to validate its therapeutic efficacy and safety.
Competing Interests: Declarations. Ethics approval and consent to participate: The study was conducted according to the guidelines of the Declaration of Helsinki. This study was approved by the Institutional Bioethics Committee of the University of Cantabria (Reference 2015/21; Project Title: Enhancing Mesenchymal Stem Cells Osteogenic Capacity Through Modulation of the Bone Marrow Microenvironment; date of approval 04–11-22) and the Consejería de Agricultura y Ganadería de Cantabria (Reference PI-08–21; Project Title: Modulation of the Regenerative Capacity of Mesenchymal Stem Cells for Their Use in Therapies Applied to the Musculoskeletal System; date of approval 01–11-21). Bone samples were obtained from the patients after anonymization. Informed consent was obtained from all the subjects involved in the study. Consent for publication: Not applicable. Competing interests: The authors declare that they have no competing interests.
(© 2025. The Author(s).)

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PREVAL19/02 Instituto de Investigación Marqués de Valdecilla; R01 CA209882 United States CA NCI NIH HHS; R37-CA251763 National Institute of Health (US); PREVAL 20/01 Instituto de Investigación Marqués de Valdecilla; NEXT-VAL Instituto de Investigación Marqués de Valdecilla; PI22/0264 Instituto de Salud Carlos III; PID2021-127493OB-C21 Ministerio de Economía y Competitividad; R01-CA209882 National Institute of Health (US)

Keywords: Smurf1; Mesenchymal stem cells; Osteoporosis; Secretome

EC 2.3.2.27 (Ubiquitin-Protein Ligases)
EC 2.3.2.26 (SMURF1 protein, human)
EC 2.3.2.26 (Smurf1 protein, rat)
0 (Culture Media, Conditioned)

Date Created: 20250207 Date Completed: 20250208 Latest Revision: 20250210

20250210

PMC11806587

10.1186/s13287-025-04165-0

39920824