{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,11,6]],"date-time":"2026-11-06T22:12:33Z","timestamp":1794003153882,"version":"4.8.0"},"reference-count":26,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2025,12,20]],"date-time":"2025-12-20T00:00:00Z","timestamp":1766188800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100011033","name":"Spanish State Research Agency","doi-asserted-by":"crossref","award":["RTI2018-101506-B-C32"],"award-info":[{"award-number":["RTI2018-101506-B-C32"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100011033","name":"Spanish State Research Agency","doi-asserted-by":"crossref","award":["RTI2018-101506-B-C31"],"award-info":[{"award-number":["RTI2018-101506-B-C31"]}],"id":[{"id":"10.13039\/501100011033","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Department of Surgery, Medical and Social Sciences, Faculty of Medicine and Health Sciences, University of Alcal\u00e1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nanomaterials"],"abstract":"<jats:p>(1) Background: Intra-articular injection is a fundamental technique in preclinical research for evaluating therapeutics and inducing joint disease models in rodents. However, the absence of standardized and validated protocols compromises reproducibility and translational validity. (2) Methods: This study establishes and experimentally validates a refined protocol for precise intra-articular injection in the knee of adult male Wistar rats. The comprehensive procedure specifies anatomical landmarks (medial border of the patellar tendon), instrumentation (27 G needle, 100 \u00b5L Hamilton syringe), a maximum volume of 35 \u00b5L, and operative verification criteria based on tactile feedback. Experimental validation was performed by administering a suspension of wear particles (2.35 mg\/mL) generated from tribocorrosion tests of CoCr surfaces biofunctionalized with graphene oxide-hyaluronic acid (GO-HA) into the left knee of five rats. (3) Results: Histological analysis using the cutting\u2013grinding technique and Toluidine Blue staining confirmed the exclusive intra-articular localization of particles in all injected animals (5\/5 success rate). Qualitative assessment revealed abundant particulate distribution within the synovial space, with numerous individual particles and multiple aggregates observed per high-power field, without evidence of extravasation in any case. (4) Conclusions: The protocol demonstrated high intra-operator repeatability and provides a reliable, ethically refined tool for precise intra-articular administration of nanomaterials and for generating robust joint disease models, thereby enhancing reproducibility and animal welfare in preclinical research.<\/jats:p>","DOI":"10.3390\/nano16010009","type":"journal-article","created":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T03:06:02Z","timestamp":1766718362000},"page":"9","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Intra-Articular Injection in Wistar Rats: Standardization and Experimental Validation of a Precise Protocol for Nanomaterial Administration"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-1049-5102","authenticated-orcid":false,"given":"Manuel","family":"Flores-S\u00e1enz","sequence":"first","affiliation":[{"name":"University of Alcal\u00e1, Department of Surgery, Medical and Social Sciences, Teaching Unit of Human Anatomy and Embryology, Laboratory of Osseointegration and Microscopic Anatomy, Science and Technology Campus, Madrid-Barcelona Highway, Km. 33.600, 28805 Alcal\u00e1 de Henares, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8697-6298","authenticated-orcid":false,"given":"Bel\u00e9n","family":"Chico","sequence":"additional","affiliation":[{"name":"Centro Nacional de Investigaciones Metal\u00fargicas (CENIM), Consejo Superior de Investigaciones Cient\u00edficas (CSIC), Avenida Gregorio del Amo, 8, 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0275-4626","authenticated-orcid":false,"given":"Maria C.","family":"Garc\u00eda Alonso","sequence":"additional","affiliation":[{"name":"Centro Nacional de Investigaciones Metal\u00fargicas (CENIM), Consejo Superior de Investigaciones Cient\u00edficas (CSIC), Avenida Gregorio del Amo, 8, 28040 Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Celia","family":"Clemente de Arriba","sequence":"additional","affiliation":[{"name":"University of Alcal\u00e1, Department of Surgery, Medical and Social Sciences, Teaching Unit of Human Anatomy and Embryology, Laboratory of Osseointegration and Microscopic Anatomy, Science and Technology Campus, Madrid-Barcelona Highway, Km. 33.600, 28805 Alcal\u00e1 de Henares, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2188-3618","authenticated-orcid":false,"given":"Soledad","family":"Aguado-Henche","sequence":"additional","affiliation":[{"name":"University of Alcal\u00e1, Department of Surgery, Medical and Social Sciences, Teaching Unit of Human Anatomy and Embryology, Laboratory of Osseointegration and Microscopic Anatomy, Science and Technology Campus, Madrid-Barcelona Highway, Km. 33.600, 28805 Alcal\u00e1 de Henares, Madrid, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"R10","DOI":"10.1186\/ar4436","article-title":"Intra-Articular Injection of Collagenase in the Knee of Rats as an Alternative Model to Study Nociception Associated with Osteoarthritis","volume":"16","author":"Santos","year":"2014","journal-title":"Arthritis Res. Ther."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"734","DOI":"10.1007\/s00586-008-0594-0","article-title":"Intra-Articular Injection of Collagenase Induced Experimental Osteoarthritis of the Lumbar Facet Joint in Rats","volume":"17","author":"Yeh","year":"2008","journal-title":"Eur. Spine J."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"273","DOI":"10.1084\/jem.68.2.273","article-title":"Sensitization and Antibody Formation with Increased Resistance to Tuberculous Infection Induced by Heat Killed Tubercle Bacilli","volume":"68","author":"Freund","year":"1938","journal-title":"J. Exp. Med."},{"key":"ref_4","unstructured":"Z\u00fa\u00f1iga, J.M., and Orellana Muriana, J.M. (2022). Ciencia y Tecnolog\u00eda en Investigaci\u00f3n Animal [Libro Interactivo], Editorial Universidad de Alcal\u00e1, SECAL (Sociedad Espa\u00f1ola para las Ciencias del Animal de Laboratorio)."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"103482","DOI":"10.1016\/j.drudis.2022.103482","article-title":"Intra-articular nanodrug delivery strategies for treating osteoarthritis","volume":"28","author":"Zhou","year":"2023","journal-title":"Drug Discov. Today"},{"key":"ref_6","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"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Escudero, M.L., Garc\u00eda-Alonso, M.C., Chico, B., Lozano, R.M., S\u00e1nchez-L\u00f3pez, L., Flores-S\u00e1enz, M., Crist\u00f3bal-Aguado, S., Moreno-G\u00f3mez-Toledano, R., and Aguado-Henche, S. (2024). Hematological Response to Particle Debris Generated During Wear\u2013Corrosion Processes of CoCr Surfaces Modified with Graphene Oxide and Hyaluronic Acid for Joint Prostheses. Nanomaterials, 14.","DOI":"10.3390\/nano14221815"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"500","DOI":"10.5694\/j.1326-5377.1960.tb73127.x","article-title":"The Principles of Humane Experimental Technique","volume":"1","author":"Russell","year":"1960","journal-title":"Med. J. Aust."},{"key":"ref_9","unstructured":"Official Journal of the European Union (2025, November 08). COMMISSION DELEGATED DIRECTIVE (EU) 2024\/1262 of 13 March 2024 Amending Directive 2010\/63\/EU of the European Parliament and of the Council as Regards the Requirements for Establishments and for the Care and Accommodation of Animals, and as Regards the Methods of Killing Animals; 2024; p. 8. Available online: https:\/\/eur-lex.europa.eu\/legal-content\/EN\/TXT\/PDF\/?uri=OJ:L_202401262."},{"key":"ref_10","unstructured":"Ministerio de la Presidencia (2025, November 08). Real Decreto 53\/2013, de 1 de Febrero, Por El Que Se Establecen Las Normas B\u00e1sicas Aplicables Para La Protecci\u00f3n de Los Animales Utilizados En Experimentaci\u00f3n y Otros Fines Cient\u00edficos, Incluyendo La Docencia; 2013; p. 48. Available online: https:\/\/www.boe.es\/eli\/es\/rd\/2013\/02\/01\/53\/con."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Wolfensohn, S., and Lloyd, M. (2003). Handbook of Laboratory Animal Management and Welfare, Wiley. [1st ed.].","DOI":"10.1002\/9780470751077"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"397","DOI":"10.1016\/S1090-3801(02)00140-4","article-title":"Evaluation of a Short Duration Behaviour-based Post-operative Pain Scoring System in Rats","volume":"7","author":"Roughan","year":"2003","journal-title":"Eur. J. Pain"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"419","DOI":"10.1177\/00236772221097472","article-title":"A Review of Methods Used to Kill Laboratory Rodents: Issues and Opportunities","volume":"56","author":"Clarkson","year":"2022","journal-title":"Lab. Anim."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"rbab020","DOI":"10.1093\/rb\/rbab020","article-title":"A pH-Responsive Mesoporous Silica Nanoparticles-Based Drug Delivery System with Controlled Release of Andrographolide for OA Treatment","volume":"8","author":"He","year":"2021","journal-title":"Regen. Biomater."},{"key":"ref_15","first-page":"614","article-title":"Administration of Substances to Laboratory Animals: Equipment Considerations, Vehicle Selection, and Solute Preparation","volume":"50","author":"Turner","year":"2011","journal-title":"J. Am. Assoc. Lab. Anim. Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.1016\/j.joca.2015.03.036","article-title":"Call for Standardized Definitions of Osteoarthritis and Risk Stratification for Clinical Trials and Clinical Use","volume":"23","author":"Kraus","year":"2015","journal-title":"Osteoarthr. Cartil."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1038\/nmeth.4462","article-title":"The Experimental Design Assistant","volume":"14","author":"Bamsey","year":"2017","journal-title":"Nat. Methods"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1111\/j.1600-0714.1982.tb00172.x","article-title":"A Method for the Study of Undecalcified Bones and Teeth with Attached Soft Tissues*: The S\u00e4ge-Schliff (Sawing and Grinding) Technique","volume":"11","author":"Donath","year":"1982","journal-title":"J. Oral Pathol. Med."},{"key":"ref_19","unstructured":"Flores S\u00e1enz, M. (2025). An\u00e1lisis Hematol\u00f3gico y Tisular de Restos de Desgaste Durante el uso de COCR Funcionalizado con \u00d3xido de Grafeno Reducido Para Aplicaciones de Pr\u00f3tesis Articulares; Respuesta In Vivo, Universidad de Alcal\u00e1. Available online: http:\/\/hdl.handle.net\/10017\/65740."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1801","DOI":"10.2217\/nnm.12.60","article-title":"Effect of Size and Dose on The Biodistribution of Graphene Oxide in Mice","volume":"7","author":"Liu","year":"2012","journal-title":"Nanomedicine"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2004","DOI":"10.1002\/jor.25245","article-title":"Assessment of Osteoarthritis Functional Outcomes and Intra-articular Injection Volume in the Rat Anterior Cruciate Ligament Transection Model","volume":"40","author":"Wang","year":"2022","journal-title":"J. Orthop. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"106658","DOI":"10.1016\/j.vascn.2019.106658","article-title":"Evaluation of Different Intraarticular Injection Volumes to Assess Optimum Efficient Amount; an Experimental Study in Rat Knee Joints","volume":"101","author":"Aytekin","year":"2020","journal-title":"J. Pharmacol. Toxicol. Methods"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.1016\/j.joca.2016.02.005","article-title":"Monoiodoacetic Acid Induces Arthritis and Synovitis in Rats in a Dose- and Time-Dependent Manner: Proposed Model-Specific Scoring Systems","volume":"24","author":"Udo","year":"2016","journal-title":"Osteoarthr. Cartil."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Takahashi, I., Matsuzaki, T., Kuroki, H., and Hoso, M. (2018). Induction of Osteoarthritis by Injecting Monosodium Iodoacetate into the Patellofemoral Joint of an Experimental Rat Model. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0196625"},{"key":"ref_25","first-page":"60649","article-title":"Osteoarthritis Pain Model Induced by Intra-Articular Injection of Mono-Iodoacetate in Rats","volume":"159","author":"Xu","year":"2020","journal-title":"J. Vis. Exp."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"93","DOI":"10.22203\/eCM.v048a06","article-title":"Who drives the progress of osteoarthritis?\u2014A descriptive study of synovium-meniscus crosstalk","volume":"48","author":"Yu","year":"2024","journal-title":"Eur. Cells Mater."}],"container-title":["Nanomaterials"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-4991\/16\/1\/9\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,12,26]],"date-time":"2025-12-26T05:17:22Z","timestamp":1766726242000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-4991\/16\/1\/9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,12,20]]},"references-count":26,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["nano16010009"],"URL":"https:\/\/doi.org\/10.3390\/nano16010009","relation":{},"ISSN":["2079-4991"],"issn-type":[{"value":"2079-4991","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,12,20]]}}}