{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T20:11:51Z","timestamp":1792181511888,"version":"4.4.0"},"reference-count":12,"publisher":"Springer Science and Business Media LLC","issue":"8","license":[{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"Medizinische Hochschule Hannover (MHH)"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Mol Cell Biochem"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Supplements are commonly used by osteoarthritis (OA) patients, and eggshell membrane (ESM) proteins are widely advertised as natural bioactive compounds for joint health. Here, we evaluated the solubility, digestibility, and immunomodulatory potential of commercial ESM supplements under physiologically relevant conditions to assess their plausibility as anti-inflammatory agents. We analyzed three commercial ESM formulations (NEM\n                    <jats:sup>\u00ae<\/jats:sup>\n                    , Torolis\n                    <jats:sup>\u00ae<\/jats:sup>\n                    , and Biova\n                    <jats:sup>\u00ae<\/jats:sup>\n                    ) using standardized extraction protocols. Protein solubility was assessed in physiological saline and under simulated gastric (pH 2) and intestinal (pH 5.8) conditions with controlled temperature, agitation, and digestive enzyme exposure. We examined the role of disulfide-crosslinked proteins in insolubility using dithiothreitol (DTT) and evaluated digestibility of proteins by SDS\u2013PAGE after treatment with pepsin, pancrelipase (Creon\n                    <jats:sup>\u00ae<\/jats:sup>\n                    ), or sequential digestion. Released peptides were identified by MALDI-TOF mass spectrometry, and immunomodulatory effects were tested using human peripheral blood mononuclear cells (PBMC) by measuring cytokine release after stimulation with lipopolysaccharide or Phorbol 12-myristate 13-acetate\/Ionomycin, in the presence or absence of Biova\n                    <jats:sup>\u00ae<\/jats:sup>\n                    . NEM\n                    <jats:sup>\u00ae<\/jats:sup>\n                    and Torolis\n                    <jats:sup>\u00ae<\/jats:sup>\n                    showed extremely low solubility (&lt;\u20090.3%) and remained insoluble under all conditions, including after DTT treatment. Although Biova\n                    <jats:sup>\u00ae<\/jats:sup>\n                    was highly soluble (~\u200990%), simulated gastrointestinal digestion produced minimal peptide generation. Only non-physiological extraction yielded sporadic trace peptides, most lacking known anti-inflammatory relevance. In PBMC assays, Biova\n                    <jats:sup>\u00ae<\/jats:sup>\n                    increased IL-6 after lipopolysaccharide stimulation, showed a non-significant TNF\u03b1 reduction, and had no effect on IL-17\u00a0A or granzyme B. Together, commercial ESM supplements are largely insoluble, resistant to digestion, and show no meaningful anti-inflammatory activity in human immune cells. These findings challenge the plausibility of orally administered ESM as a therapeutic intervention for osteoarthritis.\n                  <\/jats:p>","DOI":"10.1007\/s11010-026-05608-9","type":"journal-article","created":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T14:35:54Z","timestamp":1782916554000},"page":"3497-3507","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Clinical plausibility of eggshell membrane proteins for osteoarthritis: a study of solubility, digestibility, and immune response"],"prefix":"10.1007","volume":"481","author":[{"given":"Simona","family":"Gegel","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Arun","family":"Kumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lilia","family":"Goudeva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1558-9519","authenticated-orcid":false,"given":"J\u00fcrgen","family":"Borlak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,7,1]]},"reference":[{"key":"5608_CR1","doi-asserted-by":"publisher","first-page":"e508","DOI":"10.1016\/S2665-9913(23)00163-7","volume":"5","author":"GBD 2021 Osteoarthritis Collaborators","year":"2023","unstructured":"GBD 2021 Osteoarthritis Collaborators (2023) Global, regional, and national burden of osteoarthritis, 1990\u20132020 and projections to 2050: a systematic analysis for the Global Burden of Disease Study 2021. Lancet Rheumatol 5:e508\u2013e522. https:\/\/doi.org\/10.1016\/S2665-9913(23)00163-7","journal-title":"Lancet Rheumatol"},{"key":"5608_CR2","doi-asserted-by":"publisher","first-page":"237","DOI":"10.1016\/j.ctcp.2014.08.002","volume":"20","author":"M Basedow","year":"2014","unstructured":"Basedow M, Runciman WB, March L, Esterman A (2014) Australians with osteoarthritis; the use of and beliefs about complementary and alternative medicines. Complement Ther Clin Pract 20:237\u2013242. https:\/\/doi.org\/10.1016\/j.ctcp.2014.08.002","journal-title":"Complement Ther Clin Pract"},{"key":"5608_CR3","doi-asserted-by":"publisher","first-page":"907","DOI":"10.1007\/s10067-009-1173-4","volume":"28","author":"KJ Ruff","year":"2009","unstructured":"Ruff KJ, Winkler A, Jackson RW, DeVore DP, Ritz BW (2009) Eggshell membrane in the treatment of pain and stiffness from osteoarthritis of the knee: a randomized, multicenter, double-blind, placebo-controlled clinical study. Clin Rheumatol 28:907\u2013914. https:\/\/doi.org\/10.1007\/s10067-009-1173-4","journal-title":"Clin Rheumatol"},{"issue":"11","key":"5608_CR4","doi-asserted-by":"publisher","DOI":"10.3390\/nu14112340","volume":"14","author":"F Canovas","year":"2022","unstructured":"Canovas F, Abellan-Ruiz MS, Garcia-Munoz AM, Luque-Rubia AJ, Victoria-Montesinos D, Perez-Pinero S, Sanchez-Macarro M, Lopez-Roman FJ (2022) Randomised Clinical Trial to Analyse the Efficacy of Eggshell Membrane to Improve Joint Functionality in Knee Osteoarthritis. Nutrients 14(11):2340. https:\/\/doi.org\/10.3390\/nu14112340","journal-title":"Nutrients"},{"key":"5608_CR5","doi-asserted-by":"publisher","first-page":"292","DOI":"10.1089\/jmf.2020.0034","volume":"24","author":"JL Kiers","year":"2021","unstructured":"Kiers JL, Bult JHF (2021) Mildly Processed Natural Eggshell Membrane Alleviates Joint Pain Associated with Osteoarthritis of the Knee: A Randomized Double-Blind Placebo-Controlled Study. J Med Food 24:292\u2013298. https:\/\/doi.org\/10.1089\/jmf.2020.0034","journal-title":"J Med Food"},{"key":"5608_CR6","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1016\/j.joim.2025.04.008","volume":"23","author":"C Chen","year":"2025","unstructured":"Chen C, Wen L, Yang F, Deng Y, Ji J, Chen R, Chen Z, Chen G, Gu J (2025) Effects of dietary supplements on patients with osteoarthritis: A systematic review and network meta-analysis. J Integr Med 23:357\u2013369. https:\/\/doi.org\/10.1016\/j.joim.2025.04.008","journal-title":"J Integr Med"},{"issue":"16","key":"5608_CR7","doi-asserted-by":"publisher","DOI":"10.3390\/nu16162640","volume":"16","author":"AM Garcia-Munoz","year":"2024","unstructured":"Garcia-Munoz AM, Abellan-Ruiz MS, Garcia-Guillen AI, Victoria-Montesinos D (2024) Efficacy of Eggshell Membrane in Knee Osteoarthritis: A Systematic Review and Meta-Analysis. Nutrients 16(16):2640. https:\/\/doi.org\/10.3390\/nu16162640","journal-title":"Nutrients"},{"key":"5608_CR8","doi-asserted-by":"publisher","first-page":"2583","DOI":"10.1002\/pmic.200701147","volume":"8","author":"I Garaguso","year":"2008","unstructured":"Garaguso I, Borlak J (2008) Matrix layer sample preparation: an improved MALDI-MS peptide analysis method for proteomic studies. Proteomics 8:2583\u20132595. https:\/\/doi.org\/10.1002\/pmic.200701147","journal-title":"Proteomics"},{"key":"5608_CR9","doi-asserted-by":"publisher","first-page":"1795","DOI":"10.1002\/elps.200305387","volume":"24","author":"I Lehner","year":"2003","unstructured":"Lehner I, Niehof M, Borlak J (2003) An optimized method for the isolation and identification of membrane proteins. Electrophoresis 24:1795\u20131808. https:\/\/doi.org\/10.1002\/elps.200305387","journal-title":"Electrophoresis"},{"key":"5608_CR10","doi-asserted-by":"publisher","first-page":"9888","DOI":"10.1021\/acs.jafc.5b04266","volume":"63","author":"S Makkar","year":"2015","unstructured":"Makkar S, Liyanage R, Kannan L, Packialakshmi B, Lay JOJ, Rath NC (2015) Chicken egg shell membrane associated proteins and peptides. J Agric Food Chem 63:9888\u20139898. https:\/\/doi.org\/10.1021\/acs.jafc.5b04266","journal-title":"J Agric Food Chem"},{"key":"5608_CR11","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/j.jprot.2017.01.002","volume":"155","author":"TAE Ahmed","year":"2017","unstructured":"Ahmed TAE, Suso H, Hincke MT (2017) In-depth comparative analysis of the chicken eggshell membrane proteome. J Proteomics 155:49\u201362. https:\/\/doi.org\/10.1016\/j.jprot.2017.01.002","journal-title":"J Proteomics"},{"key":"5608_CR12","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2023.1116126","volume":"10","author":"C Han","year":"2023","unstructured":"Han C, Chen Y, Shi L, Chen H, Li L, Ning Z, Zeng D, Wang D (2023) Advances in eggshell membrane separation and solubilization technologies. Front Vet Sci 10:1116126. https:\/\/doi.org\/10.3389\/fvets.2023.1116126","journal-title":"Front Vet Sci"}],"container-title":["Molecular and Cellular Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11010-026-05608-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s11010-026-05608-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s11010-026-05608-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,28]],"date-time":"2026-08-28T09:14:48Z","timestamp":1787908488000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s11010-026-05608-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,1]]},"references-count":12,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["5608"],"URL":"https:\/\/doi.org\/10.1007\/s11010-026-05608-9","relation":{},"ISSN":["1573-4919"],"issn-type":[{"value":"1573-4919","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,1]]},"assertion":[{"value":"18 March 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 June 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 July 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The study was approved by the Ethics Committee of Hannover Medical School.","order":2,"name":"Ethics","label":"Ethical approval","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"Written informed consent was obtained from all donors.","order":3,"name":"Ethics","label":"Consent to participate","group":{"name":"EthicsHeading","label":"Declarations"}}]}}