{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,20]],"date-time":"2026-03-20T02:19:50Z","timestamp":1773973190090,"version":"3.50.1"},"reference-count":29,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,22]],"date-time":"2025-09-22T00:00:00Z","timestamp":1758499200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Activator MAF, LLC"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Veterinary Sciences"],"abstract":"<jats:p>Osteoarthritis is a prevalent and disabling condition in companion dogs, yet existing treatments are primarily symptomatic and limited by safety concerns. EF-M2, a defined derivative of vitamin D-binding protein, selectively biases macrophages toward an anti-inflammatory phenotype in vitro. We conducted a randomised, double-blind, placebo-controlled trial (IMPAWS-OA-1) in 60 client-owned dogs with naturally occurring hip or elbow osteoarthritis. Animals were allocated to subcutaneous EF-M2 (0.1 \u00b5g\/kg) given thrice weekly or twice weekly, or to saline placebo for four weeks, followed by four weeks off-drug. The primary endpoint was change in Canine Brief Pain Inventory\u2013Pain Severity Score (CBPI-PSS) at Day 28. EF-M2 produced dose\u2013frequency-dependent benefits: LS-mean \u0394PSS was \u22122.11 for thrice weekly, \u22121.42 for twice weekly, and \u22120.54 for placebo (arm effect p &lt; 0.001). Objective measures showed parallel improvements in peak vertical force and accelerometery. Serum biomarkers confirmed macrophage repolarisation (ARG1\/iNOS ratio, IL-10 increase, TNF-\u03b1 decrease), correlating with clinical response. Adverse events were infrequent and mild, with no excess over placebo. In conclusion, EF-M2 achieved clinically meaningful pain relief, functional gains, and biomarker shifts without safety signals, establishing first-in-species proof that targeted macrophage modulation may be a viable disease-modifying approach for canine osteoarthritis.<\/jats:p>","DOI":"10.3390\/vetsci12090919","type":"journal-article","created":{"date-parts":[[2025,9,22]],"date-time":"2025-09-22T17:08:17Z","timestamp":1758560897000},"page":"919","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Targeted Macrophage Modulation as a Disease-Modifying Approach in Canine Osteoarthritis: The Efficacy of EF-M2 (ImmutalonTM) in a Double-Blind Placebo-Controlled Study"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9494-4234","authenticated-orcid":false,"given":"Evgeny","family":"Pokushalov","sequence":"first","affiliation":[{"name":"Scientific Research Laboratory, Triangel Scientific, San Francisco, CA 94101, USA"},{"name":"Center for New Medical Technologies, 630090 Novosibirsk, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1878-4467","authenticated-orcid":false,"given":"Dmitry","family":"Kudlay","sequence":"additional","affiliation":[{"name":"Institute of Pharmacy, I.M. Sechenov First Moscow State Medical University (Sechenov University), 119435 Moscow, Russia"},{"name":"Institute of Medicine and Medical Technologies, Novosibirsk State University, 630090 Novosibirsk, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nikolai","family":"Revkov","sequence":"additional","affiliation":[{"name":"VEGA Veterinary Clinic, 630049 Novosibirsk, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anastasya","family":"Shcherbakova","sequence":"additional","affiliation":[{"name":"BALTO Veterinary Clinic, 630055 Novosibirsk, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael","family":"Johnson","sequence":"additional","affiliation":[{"name":"Scientific Research Laboratory, Triangel Scientific, San Francisco, CA 94101, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard","family":"Miller","sequence":"additional","affiliation":[{"name":"Scientific Research Laboratory, Triangel Scientific, San Francisco, CA 94101, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Anderson, K.L., Zulch, H., O\u2019Neill, D.G., Meeson, R.L., and Collins, L.M. (2020). Risk Factors for Canine Osteoarthritis and Its Predisposing Arthropathies: A Systematic Review. Front. Vet. Sci., 7.","DOI":"10.3389\/fvets.2020.00220"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/j.prevetmed.2016.07.017","article-title":"The attitudes of owners and veterinary professionals in the United Kingdom to the risk of adverse events associated with using non-steroidal anti-inflammatory drugs (NSAIDs) to treat dogs with osteoarthritis","volume":"131","author":"Belshaw","year":"2016","journal-title":"Prev. Vet. Med."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Monteiro, B.P., Simon, A., Knesl, O., Mandello, K., Nederveld, S., Olby, N.J., Innes, J.F., and Lascelles, B.D.X. (2025). Global pharmacovigilance reporting of the first monoclonal antibody for canine osteoarthritis: A case study with bedinvetmab (Librela\u2122). Front. Vet. Sci., 12.","DOI":"10.3389\/fvets.2025.1558222"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"853","DOI":"10.1111\/jvim.16057","article-title":"Prevalence of gastrointestinal lesions in dogs chronically treated with nonsteroidal anti-inflammatory drugs","volume":"35","author":"Mabry","year":"2021","journal-title":"J. Vet. Intern. Med."},{"key":"ref_5","unstructured":"US Food and Drug Administration, Center for Veterinary Medicine (2025, August 06). Get the Facts About Pain Relievers for Pets [Internet], Available online: https:\/\/www.fda.gov\/animal-veterinary\/animal-health-literacy\/get-facts-about-pain-relievers-pets."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Carreira, L.M., Cota, J., Alves, J., In\u00e1cio, F., Alexandre-Pires, G., and Azevedo, P. (2024). A Pilot Study of the Clinical Effectiveness of a Single Intra-Articular Injection of Stanozolol in Canines with Knee Degenerative Joint Disease and Its Correlation with Serum Interleukin-1\u03b2 Levels. Animals, 14.","DOI":"10.3390\/ani14091351"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"379","DOI":"10.1016\/j.rvsc.2012.11.020","article-title":"Effects of intraarticular treatment with stanozolol on synovial membrane and cartilage in an ovine model of osteoarthritis","volume":"94","author":"Spadari","year":"2013","journal-title":"Res. Vet. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1111\/j.1939-1676.2007.0014.x","article-title":"Amantadine in a multimodal analgesic regimen for alleviation of refractory osteoarthritis pain in dogs","volume":"22","author":"Lascelles","year":"2008","journal-title":"J. Vet. Intern. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1136\/vr.m1594","article-title":"Does the addition of cannabidiol alongside current drug treatments reduce pain in dogs with osteoarthritis?","volume":"186","author":"Morrow","year":"2020","journal-title":"Vet. Rec."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1002\/art.39842","article-title":"Review: Metabolic Regulation of Inflammation in Osteoarthritis","volume":"69","author":"Berenbaum","year":"2017","journal-title":"Arthritis Rheumatol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1038\/s41584-019-0202-1","article-title":"Spontaneous dog osteoarthritis\u2014A One Medicine vision","volume":"15","author":"Meeson","year":"2019","journal-title":"Nat. Rev. Rheumatol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1111\/vsu.12479","article-title":"Systematic Review of Outcome Measures Reported in Clinical Canine Osteoarthritis Research","volume":"45","author":"Belshaw","year":"2016","journal-title":"Vet. Surg."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Zhao, K., Ruan, J., Nie, L., Ye, X., and Li, J. (2023). Effects of synovial macrophages in osteoarthritis. Front. Immunol., 14.","DOI":"10.3389\/fimmu.2023.1164137"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Ma, Y., Yang, H., Zong, X., Wu, J., Ji, X., Liu, W., Yuan, P., Chen, X., Yang, C., and Li, X. (2021). Artificial M2 macrophages for disease-modifying osteoarthritis therapeutics. Biomaterials, 274.","DOI":"10.1016\/j.biomaterials.2021.120865"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Nabeshima, Y., Abe, C., Kawauchi, T., Hiroi, T., Uto, Y., and Nabeshima, Y.I. (2020). Simple method for large-scale production of macrophage activating factor GcMAF. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-75571-y"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Kirikovich, S.S., Levites, E.V., Proskurina, A.S., Ritter, G.S., Peltek, S.E., Vasilieva, A.R., Ruzanova, V.S., Dolgova, E.V., Oshihmina, S.G., and Sysoev, A.V. (2023). The Molecular Aspects of Functional Activity of Macrophage-Activating Factor GcMAF. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms242417396"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"10934","DOI":"10.3390\/cimb46100650","article-title":"Production of GcMAF with Anti-Inflammatory Properties and Its Effect on Models of Induced Arthritis in Mice and Cystitis in Rats","volume":"46","author":"Kirikovich","year":"2024","journal-title":"Curr. Issues Mol. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"eabe0083","DOI":"10.1126\/sciadv.abe0083","article-title":"Metabolically engineered stem cell-derived exosomes to regulate macrophage heterogeneity in rheumatoid arthritis","volume":"7","author":"You","year":"2021","journal-title":"Sci Adv"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Cachon, T., Frykman, O., Innes, J.F., Lascelles, B.D.X., Okumura, M., Sousa, P., Staffieri, F., Steagall, P.V., and Van Ryssen, B. (2023). COAST Development Group\u2019s international consensus guidelines for the treatment of canine osteoarthritis. Front. Vet. Sci., 10.","DOI":"10.3389\/fvets.2023.1137888"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Innes, J.F., Morton, M.A., and Lascelles, B.D.X. (2023). Minimal clinically-important differences for the \u2018Liverpool Osteoarthritis in Dogs\u2019 (LOAD) and the \u2018Canine Orthopedic Index\u2019 (COI) client-reported outcomes measures. PLoS ONE, 18.","DOI":"10.1371\/journal.pone.0280912"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Alves, J.C., Santos, A., Lavrador, C., and Carreira, L.M. (2023). Minimal Clinically Important Differences for a Weight Distribution Platform in Dogs with Osteoarthritis. Animals, 14.","DOI":"10.3390\/ani14010128"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1278","DOI":"10.2460\/javma.233.8.1278","article-title":"Ability of the canine brief pain inventory to detect response to treatment in dogs with osteoarthritis","volume":"233","author":"Brown","year":"2008","journal-title":"J. Am. Vet. Med. Assoc."},{"key":"ref_23","unstructured":"US Food and Drug Administration, Center for Veterinary Medicine (2025, August 06). Dear Veterinarian Letter Notifying Veterinarians About Adverse Events Reported in Dogs Treated with Librela (Bedinvetmab Injection) [Internet], Available online: https:\/\/www.fda.gov\/animal-veterinary\/product-safety-information\/dear-veterinarian-letter-notifying-veterinarians-about-adverse-events-reported-dogs-treated-librela."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Yang, Z., and Ming, X.F. (2014). Functions of arginase isoforms in macrophage inflammatory responses: Impact on cardiovascular diseases and metabolic disorders. Front. Immunol., 5.","DOI":"10.3389\/fimmu.2014.00533"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"757","DOI":"10.3892\/etm.2022.11693","article-title":"Role of macrophage polarization in osteoarthritis (Review)","volume":"24","author":"Chen","year":"2022","journal-title":"Exp. Ther. Med."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Hoober, J.K. (2020). ASGR1 and Its Enigmatic Relative, CLEC10A. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21144818"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2713","DOI":"10.2147\/JIR.S313179","article-title":"Spermidine Inhibits Joints Inflammation and Macrophage Activation in Mice with Collagen-Induced Arthritis","volume":"14","author":"Yuan","year":"2021","journal-title":"J. Inflamm. Res."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Szczykutowicz, J. (2023). Ligand Recognition by the Macrophage Galactose-Type C-Type Lectin: Self or Non-Self?-A Way to Trick the Host\u2019s Immune System. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms242317078"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Percie du Sert, N., Hurst, V., Ahluwalia, A., Alam, S., Avey, M.T., Baker, M., Browne, W.J., Clark, A., Cuthill, I.C., and Dirnagl, U. (2020). The ARRIVE guidelines 2.0: Updated guidelines for reporting animal research. PLoS Biol., 18.","DOI":"10.1371\/journal.pbio.3000410"}],"container-title":["Veterinary Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-7381\/12\/9\/919\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:47:13Z","timestamp":1760035633000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-7381\/12\/9\/919"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,9,22]]},"references-count":29,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["vetsci12090919"],"URL":"https:\/\/doi.org\/10.3390\/vetsci12090919","relation":{},"ISSN":["2306-7381"],"issn-type":[{"value":"2306-7381","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,9,22]]}}}