{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T05:31:23Z","timestamp":1767245483236,"version":"3.48.0"},"reference-count":51,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,29]],"date-time":"2025-12-29T00:00:00Z","timestamp":1766966400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100017637","name":"Wroclaw University of Environmental and Life Sciences","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100017637","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Life"],"abstract":"<jats:p>Mammary gland tumors are among the most common neoplasms in female dogs. Macrophages are believed to play an important role in tumor progression and metastasis. The aim of this study was to determine whether hemosiderin-laden macrophages (HLMs) may be involved in the development of metastatic lesions in regional lymph nodes. Forty-two cases of mammary gland cancers in female dogs and their regional lymph nodes were included in the analysis. The samples were divided into two groups based on the presence or absence of metastases. The sections were stained with hematoxylin and eosin (HE) and Prussian blue and were additionally subjected to immunohistochemical labeling using antibodies against pan-cytokeratin (Pan-CK) and Ki-67. In 20 cases, no metastatic changes were detected in the regional lymph node, whereas metastases were identified in 22 cases. A positive correlation was observed between the number of HLMs in the tumor stroma and the number of HLMs in the regional lymph node. Furthermore, a positive correlation was found between Ki-67 nuclear immunoreactivity in the mammary tumor and the number of HLMs present within its stroma. HLMs may represent a component of the tumor microenvironment that promotes cancer cell proliferation and potentially contributes to the propensity for metastasis formation in mammary tumors of female dogs.<\/jats:p>","DOI":"10.3390\/life16010053","type":"journal-article","created":{"date-parts":[[2025,12,31]],"date-time":"2025-12-31T16:34:23Z","timestamp":1767198863000},"page":"53","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["The Role of Hemosiderin-Laden Macrophages (HLMs) in the Metastasis of Mammary Gland Cancers in Bitches"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4033-4921","authenticated-orcid":false,"given":"Kacper","family":"\u017bebrowski","sequence":"first","affiliation":[{"name":"Department of Pathology, Division of Pathomorphology and Veterinary Forensics, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2198-8877","authenticated-orcid":false,"given":"Ma\u0142gorzata","family":"Kandefer-Gola","sequence":"additional","affiliation":[{"name":"Department of Pathology, Division of Pathomorphology and Veterinary Forensics, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5166-856X","authenticated-orcid":false,"given":"Izabela","family":"Janus-Zi\u00f3\u0142kowska","sequence":"additional","affiliation":[{"name":"Department of Pathology, Division of Pathomorphology and Veterinary Forensics, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rafa\u0142","family":"Ciaputa","sequence":"additional","affiliation":[{"name":"Department of Pathology, Division of Pathomorphology and Veterinary Forensics, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7203-2392","authenticated-orcid":false,"given":"Stanis\u0142aw","family":"Dzimira","sequence":"additional","affiliation":[{"name":"Department of Pathology, Division of Pathomorphology and Veterinary Forensics, Faculty of Veterinary Medicine, Wroclaw University of Environmental and Life Sciences, 50-375 Wroclaw, Poland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Brancewicz, J., W\u00f3jcik, N., Sarnowska, Z., Robak, J., and Kr\u00f3l, M. (2025). The Multifaceted Role of Macrophages in Biology and Diseases. Int. J. Mol. Sci., 26.","DOI":"10.3390\/ijms26052107"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"8203","DOI":"10.2174\/0929867328666210720143721","article-title":"The role of tumor associated macrophages (TAMs) in cancer progression, chemoresistance, angiogenesis and metastasis\u2014Current status","volume":"28","author":"Dallavalasa","year":"2021","journal-title":"Curr. Med. Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"737","DOI":"10.1038\/35036374","article-title":"Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis","volume":"2","author":"Bergers","year":"2000","journal-title":"Nat. Cell Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"587","DOI":"10.3892\/ijo.2014.2761","article-title":"Macrophage-secreted IL-8 induces epithelial-mesenchymal transition in hepatocellular carcinoma cells by activating the JAK2\/STAT3\/Snail pathway","volume":"46","author":"Fu","year":"2015","journal-title":"Int. J. Oncol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1038\/nrc2618","article-title":"Microenvironmental regulation of metastasis","volume":"9","author":"Joyce","year":"2009","journal-title":"Nat. Rev. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"11238","DOI":"10.1158\/0008-5472.CAN-06-1278","article-title":"Macrophages regulate the angiogenic switch in a mouse model of breast cancer","volume":"66","author":"Lin","year":"2006","journal-title":"Cancer Res."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Qian, B., Deng, Y., Im, J.H., Muschel, R.J., Zou, Y., Li, J., Lang, R.A., and Pollard, J.W. (2009). A distinct macrophage population mediates metastatic breast cancer cell extravasation, establishment and growth. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0006562"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1007\/s10555-013-9420-1","article-title":"The pre-metastatic niche: Finding common ground","volume":"32","author":"Sceneay","year":"2013","journal-title":"Cancer Metastasis Rev."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1016\/j.ccr.2014.03.021","article-title":"A positive feedback loop between Mesenchymal- like cancer cells and macrophages is essential to breast cancer metastasis","volume":"25","author":"Su","year":"2014","journal-title":"Cancer Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"5242","DOI":"10.1158\/0008-5472.CAN-05-4463","article-title":"Tumor cell derived and macrophage derived cathepsin B promotes progression and lung metastasis of mammary cancer","volume":"66","author":"Vasiljeva","year":"2006","journal-title":"Cancer Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7022","DOI":"10.1158\/0008-5472.CAN-04-1449","article-title":"A paracrine loop between tumor cells and macrophages is required for tumor cell migration in mammary tumors","volume":"64","author":"Wyckoff","year":"2004","journal-title":"Cancer Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"799","DOI":"10.1038\/s41573-022-00520-5","article-title":"Macrophages as Tools and Targets in Cancer Therapy","volume":"21","author":"Mantovani","year":"2022","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1111\/j.1600-065X.2008.00607.x","article-title":"The Yin-Yang of tumor-associated macrophages in neoplastic progression and immune surveillance","volume":"222","author":"Allavena","year":"2008","journal-title":"Immunol. Rev."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2011","DOI":"10.4049\/jimmunol.180.4.2011","article-title":"Plasticity of macrophage function during tumor progression: Regulation by distinct molecular mechanisms","volume":"180","author":"Biswas","year":"2008","journal-title":"J. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"39","DOI":"10.1016\/j.cell.2010.03.014","article-title":"Macrophage diversity enhances tumor progression and metastasis","volume":"141","author":"Qian","year":"2010","journal-title":"Cell"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1172\/JCI59643","article-title":"Macrophage plasticity and polarization: In Vivo veritas","volume":"122","author":"Sica","year":"2012","journal-title":"J. Clin. Investig."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"295","DOI":"10.1016\/j.imbio.2014.09.011","article-title":"Macrophage iron homeostasis and polarization in the context of cancer","volume":"220","author":"Jung","year":"2015","journal-title":"Immunobiology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1336","DOI":"10.1039\/C4MT00104D","article-title":"Macrophages: Central regulators of iron balance","volume":"6","author":"Gammella","year":"2014","journal-title":"Metallomics"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"500","DOI":"10.1038\/nri3863","article-title":"Iron homeostasis in host defence and inflammation","volume":"15","author":"Ganz","year":"2015","journal-title":"Nat. Rev. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1016\/j.autrev.2012.03.003","article-title":"Iron levels in polarized macrophages: Regulation of immunity and autoimmunity","volume":"11","author":"Recalcati","year":"2012","journal-title":"Autoimmun. Rev."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1016\/j.it.2011.03.007","article-title":"Iron trafficking and metabolism in macrophages: Contribution to the polarized phenotype","volume":"32","author":"Cairo","year":"2011","journal-title":"Trends Immunol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"330","DOI":"10.1177\/0300985811409253","article-title":"A Retrospective Study of Those Histopathologic Parameters Predictive of Invasion of the Lymphatic System by Canine Mammary Carcinomas","volume":"49","author":"Rasotto","year":"2012","journal-title":"Vet. Pathol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Zheng, H.H., Du, C.T., Yu, C., Zhang, Y.Z., Huang, R.L., Tang, X.Y., and Xie, G.H. (2022). Epidemiological Investigation of Canine Mammary Tumors in Mainland China Between 2017 and 2021. Front. Vet. Sci., 22.","DOI":"10.3389\/fvets.2022.843390"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1714","DOI":"10.1590\/1678-4162-10217","article-title":"Epidemiological, clinical and pathological evaluation of overall survival in canines with mammary neoplasms","volume":"70","author":"Nunes","year":"2018","journal-title":"Arq. Bras. Med. Vet. Zootec."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1016\/S0195-5616(03)00020-2","article-title":"Canine mammary gland tumors","volume":"33","author":"Sorenmo","year":"2003","journal-title":"Vet. Clin. N. Am. Small Anim. Pract."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"117","DOI":"10.1177\/0300985810393258","article-title":"Classification and grading of canine mammary tumors","volume":"48","author":"Goldschmidt","year":"2011","journal-title":"Vet. Pathol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1111\/j.1365-2559.1991.tb00229.x","article-title":"Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: Experience from a large study with long-term follow-up","volume":"19","author":"Elston","year":"1991","journal-title":"Histopathology"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"94","DOI":"10.1177\/0300985812447830","article-title":"Prognostic value of histological grading in noninflammatory canine mammary carcinomas in a prospective study with two-year follow-up: Relationship with clinical and histological characteristics","volume":"50","author":"Clemente","year":"2013","journal-title":"Vet Pathol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.tjem.2018.08.001","article-title":"User\u2019s guide to correlation coefficients","volume":"7","author":"Akoglu","year":"2018","journal-title":"Turk. J. Emerg. Med."},{"key":"ref_30","first-page":"523","article-title":"Haemosiderin-laden macrophages (HLMs): Current understanding and clinical relevance","volume":"81","author":"Dzimira","year":"2025","journal-title":"Med. Weter."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Armando, F., Godizzi, F., Razzuoli, E., Leonardi, F., Angelone, M., Corradi, A., Meloni, D., Ferrari, L., and Passeri, B. (2020). Epithelial to Mesenchymal Transition (EMT) in a Laryngeal Squamous Cell Carcinoma of a Horse: Future Perspectives. Animals, 10.","DOI":"10.3390\/ani10122318"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"622","DOI":"10.1177\/0300985818776054","article-title":"Characteristics of the Epithelial-Mesenchymal Transition in Primary and Paired Metastatic Canine Mammary Carcinomas","volume":"55","author":"Brisson","year":"2018","journal-title":"Vet. Pathol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Armando, F., Mazzola, F., Ferrari, L., and Corradi, A. (2023). An Overview of Epithelial-to-Mesenchymal Transition and Mesenchymal-to-Epithelial Transition in Canine Tumors: How Far Have We Come?. Vet. Sci., 10.","DOI":"10.3390\/vetsci10010019"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"824","DOI":"10.1002\/eji.200939889","article-title":"Differential regulation of iron homeostasis during human macrophage polarized activation","volume":"40","author":"Recalcati","year":"2010","journal-title":"Eur. J. Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1016\/j.freeradbiomed.2010.12.028","article-title":"Effects of iron deficiency and iron overload on angiogenesis and oxidative stress-a potential dual role for iron in breast cancer","volume":"1","author":"Jian","year":"2011","journal-title":"Free Radic. Biol. Med."},{"key":"ref_36","first-page":"262","article-title":"Referenzwerteh\u00e4mosiderin be ladener Alveo larmakrophagen in der bronchial veol\u00e4ren Lavagebei Kindern [Reference values of hemosiderin-laden alveolar macrophages in bronchial veolarlavage in children]","volume":"53","author":"Ahrens","year":"1999","journal-title":"Pneumologie"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"166","DOI":"10.1136\/bmj.2.5964.166","article-title":"Occult pulmonary haemorrhage in leukaemia","volume":"26","author":"Golde","year":"1975","journal-title":"Br. Med. J."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1164\/ajrccm\/136.1.155","article-title":"Diagnosis of pulmonary hemorrhage in the immunocompromised host","volume":"136","author":"Kahn","year":"1987","journal-title":"Am. Rev. Respir. Dis."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"157","DOI":"10.1164\/ajrccm.151.1.7812547","article-title":"Alveolar hemorrhage. Diagnostic criteria and results in 194 immunocompromised hosts","volume":"151","author":"Lassence","year":"1995","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_40","first-page":"26","article-title":"Hemosiderin-laden macrophages in broncho alveolar lavage fluid","volume":"36","year":"1992","journal-title":"Acta Cytol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"972","DOI":"10.1002\/ppul.20479","article-title":"Quantification of siderophages in broncho alveolar fluid in transfusional and primary pulmonary hemosiderosis","volume":"41","author":"Priftis","year":"2006","journal-title":"Pediatr. Pulmonol."},{"key":"ref_42","unstructured":"Raskin, R.E., and Meyer, D.J. (2016). Respiratory tract. Canine and Feline Cytology, W.B. Saunders. [3rd ed.]."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1111\/j.1939-165X.2005.tb00003.x","article-title":"Frequency and severity of tracheal wash hemosiderosis and association with underlying disease in 96 cats: 2002\u20132003","volume":"34","author":"DeHeer","year":"2005","journal-title":"Vet. Clin. Pathol."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Giambrone, G., Vullo, C., Puleio, R., Rifici, C., Di Giorgio, S., and Sfacteria, A. (2023). Hemosiderin-laden macrophages in canine mammary carcinomas. Animals, 13.","DOI":"10.3390\/ani13233634"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1016\/j.tvjl.2018.02.016","article-title":"Tumour-associated macrophages: Relation with progression and invasiveness, and assessment of M1\/M2 macrophages in canine mammary tumours","volume":"234","author":"Monteiro","year":"2018","journal-title":"Vet. J."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"104","DOI":"10.3892\/ijo.2017.3996","article-title":"Effect of macrophages on breast cancer cell proliferation, and on expression of hormone receptors, uPAR and HER-2","volume":"51","author":"Lindsten","year":"2017","journal-title":"Int. J. Oncol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1186\/s12967-025-06556-6","article-title":"Macrophages in the premetastatic and metastatic niche: Key functions and therapeutic directions","volume":"23","author":"Qi","year":"2025","journal-title":"J. Transl. Med."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1245\/s10434-014-4054-2","article-title":"Correlations between molecular subtypes and pathologic response patterns of breast cancers after neoadjuvant chemotherapy","volume":"22","author":"Lee","year":"2015","journal-title":"Ann. Surg. Oncol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"875","DOI":"10.1038\/s41379-020-00714-5","article-title":"Clinico-pathologic predictors of patterns of residual disease following neoadjuvant chemotherapy for breast cancer","volume":"34","author":"Pastorello","year":"2021","journal-title":"Mod. Pathol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1080\/10520295.2017.1369160","article-title":"Expression of iron-related proteins in feline and canine mammary gland reveals unexpected accumulation of iron","volume":"92","author":"Marques","year":"2017","journal-title":"Biotech. Histochem."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Syrtveit, A.A., Ingebriktsen, L.M., Tegnander, A.F., Akslen, L.A., Wik, E., and Hoivik, E.A. (2025). Tumor necrosis associates with aggressive breast cancer features, increased hypoxia signaling and reduced patient survival. Sci. Rep.","DOI":"10.1038\/s41598-025-29905-3"}],"container-title":["Life"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/1\/53\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T05:29:02Z","timestamp":1767245342000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2075-1729\/16\/1\/53"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,29]]},"references-count":51,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["life16010053"],"URL":"https:\/\/doi.org\/10.3390\/life16010053","relation":{},"ISSN":["2075-1729"],"issn-type":[{"type":"electronic","value":"2075-1729"}],"subject":[],"published":{"date-parts":[[2025,12,29]]}}}