{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T15:12:56Z","timestamp":1783696376835,"version":"3.55.0"},"reference-count":31,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T00:00:00Z","timestamp":1783641600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Alterations in the ceramide (CER) profile have been associated with atopic dermatitis in dogs; however, the specific effects of CER subclasses on canine keratinocytes remain unclear. This study aimed to optimize a CER complex and evaluate its effects on cytokine expression, CER synthesis, and skin barrier formation using canine epidermal keratinocyte progenitor (CPEK) cells and a reconstructed canine epidermis (RCE) model. Among various CER combinations tested, only the mixture of CER [NS] and [EOP] significantly reduced proinflammatory cytokine mRNA levels in Staphylococcus pseudintermedius\u2013stimulated CPEK cells. Further optimization revealed that a 4:1 ratio of CER [NS] to [EOP] (cCER complex) markedly decreased CCL17 and CCL22 mRNA expression while upregulating keratin 10, ceramide synthase 3 (CERS3), and ceramide synthase 4, as well as enzymes involved in both de novo and salvage CER biosynthesis pathways. In the RCE model, cCER administration improved stratum corneum organization and increased the expression of loricrin, CERS3, and CER. In vivo application of cCER in dogs accelerated skin barrier repair and improved hair condition. These findings suggest that a CER [NS]\/[EOP] mixture represents a promising therapeutic option with anti-inflammatory and skin barrier-enhancing effects for canine atopic dermatitis.<\/jats:p>","DOI":"10.3390\/ani16142148","type":"journal-article","created":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T14:14:25Z","timestamp":1783692865000},"page":"2148","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Optimization of a Canine-Specific Ceramide Complex and Evaluation of Its Effects on In Vitro and In Vivo Canine Skin"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0002-3518-384X","authenticated-orcid":false,"given":"Hee Yeon","family":"Cho","sequence":"first","affiliation":[{"name":"R&I Center, COSMAX BTI, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-1704-4230","authenticated-orcid":false,"given":"Bo-Eun","family":"Kim","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eunjin","family":"Park","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX BTI, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3522-7208","authenticated-orcid":false,"given":"Minji","family":"Kim","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX BTI, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Younhwa","family":"Nho","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX BTI, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyunna","family":"Kim","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9318-8318","authenticated-orcid":false,"given":"Seunghyun","family":"Kang","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunho","family":"Park","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX BTI, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8656-2263","authenticated-orcid":false,"given":"Kyung Eun","family":"Lee","sequence":"additional","affiliation":[{"name":"R&I Center, COSMAX BTI, Seongnam 13486, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Fujii, M. (2021). The Pathogenic and Therapeutic Implications of Ceramide Abnormalities in Atopic Dermatitis. Cells, 10.","DOI":"10.3390\/cells10092386"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"729","DOI":"10.1007\/s00403-018-1859-z","article-title":"Age-Related and Seasonal Changes in Covalently Bound Ceramide Content in Forearm Stratum Corneum of Japanese Subjects: Determination of Molecular Species of Ceramides","volume":"310","author":"Fujiwara","year":"2018","journal-title":"Arch. Dermatol. Res."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"732","DOI":"10.1111\/j.1600-0625.2011.01306.x","article-title":"Alteration of Stratum Corneum Ceramide Profiles in Spontaneous Canine Model of Atopic Dermatitis","volume":"20","author":"Yoon","year":"2011","journal-title":"Exp. Dermatol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1111\/j.1365-3164.2012.01102.x","article-title":"Skin Lipid Profiling in Normal and Seborrhoeic Shih Tzu Dogs","volume":"24","author":"Yoon","year":"2013","journal-title":"Vet. Dermatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1007\/s00403-011-1120-5","article-title":"Atopic Dermatitis in Dogs Is Associated with a High Heterogeneity in the Distribution of Protein-Bound Lipids within the Stratum Corneum","volume":"303","author":"Popa","year":"2011","journal-title":"Arch. Dermatol. Res."},{"key":"ref_6","first-page":"15","article-title":"Canine Atopic Dermatitis: Prevalence, Impact, and Management Strategies","volume":"15","author":"Drechsler","year":"2024","journal-title":"Vet. Med. Res. Rep."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"100235","DOI":"10.1016\/j.jlr.2022.100235","article-title":"Whole Picture of Human Stratum Corneum Ceramides, Including the Chain-Length Diversity of Long-Chain Bases","volume":"63","author":"Suzuki","year":"2022","journal-title":"J. Lipid Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1111\/j.1365-3164.2009.00759.x","article-title":"Characterization and Quantification of Ceramides in the Nonlesional Skin of Canine Patients with Atopic Dermatitis Compared with Controls","volume":"20","author":"Reiter","year":"2009","journal-title":"Vet. Dermatol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1007\/s11259-011-9477-7","article-title":"Reconstruction of Stratum Corneum in Organotypically Cultured Canine Keratinocyte-Derived CPEK Cells","volume":"35","author":"Yagihara","year":"2011","journal-title":"Vet. Res. Commun."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1111\/vde.12655","article-title":"Expression of ZO-1 and Claudin-1 in a 3D Epidermal Equivalent Using Canine Progenitor Epidermal Keratinocytes","volume":"29","author":"Teramoto","year":"2018","journal-title":"Vet. Dermatol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1016\/j.vetimm.2011.07.014","article-title":"Is the Skin Barrier Abnormal in Dogs with Atopic Dermatitis?","volume":"144","author":"Olivry","year":"2011","journal-title":"Vet. Immunol. Immunopathol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"105055","DOI":"10.1016\/j.chemphyslip.2021.105055","article-title":"Skin Lipids in Health and Disease: A Review","volume":"236","author":"Knox","year":"2021","journal-title":"Chem. Phys. Lipids"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1111\/vde.13159","article-title":"Cytokine Transcriptome Profiling in Acute Experimental Canine Atopic Dermatitis Skin Lesions after IL-31 Inhibition with Lokivetmab","volume":"34","author":"Crawford","year":"2023","journal-title":"Vet. Dermatol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1111\/vde.13192","article-title":"Update on the Role of Cytokines and Chemokines in Canine Atopic Dermatitis","volume":"35","author":"Santoro","year":"2024","journal-title":"Vet. Dermatol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1002\/jcp.21101","article-title":"Ceramide Inhibits LPS-induced Production of IL-5, IL-10, and IL-13 from Mast Cells","volume":"213","author":"Chiba","year":"2007","journal-title":"J. Cell. Physiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"653","DOI":"10.1007\/s12257-017-0216-y","article-title":"Anti-Inflammatory Effect of the Ceramide Mixture Extracted from Genetically Modified Saccharomyces Cerevisiae","volume":"22","author":"Song","year":"2017","journal-title":"Biotechnol. Bioprocess Eng."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1159\/000518517","article-title":"A Lipid Mixture Enriched by Ceramide NP with Fatty Acids of Diverse Chain Lengths Contributes to Restore the Skin Barrier Function Impaired by Topical Corticosteroid","volume":"35","author":"Lim","year":"2022","journal-title":"Skin. Pharmacol. Physiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"E1334","DOI":"10.1073\/pnas.1504555113","article-title":"ER Stress Stimulates Production of the Key Antimicrobial Peptide, Cathelicidin, by Forming a Previously Unidentified Intracellular S1P Signaling Complex","volume":"113","author":"Park","year":"2016","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1016\/j.jid.2018.08.015","article-title":"Both Sphingosine Kinase 1 and 2 Coordinately Regulate Cathelicidin Antimicrobial Peptide Production during Keratinocyte Differentiation","volume":"139","author":"Shin","year":"2019","journal-title":"J. Investig. Dermatol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"117985","DOI":"10.1016\/j.biopha.2025.117985","article-title":"Regulation of Keratinocyte Proliferation and Differentiation by Secoiridoid Oleacein in Monoculture and Fibroblast Co-Culture Models","volume":"185","author":"Cho","year":"2025","journal-title":"Biomed. Pharmacother."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"789","DOI":"10.1042\/BJ20111626","article-title":"Ceramide Synthases at the Centre of Sphingolipid Metabolism and Biology","volume":"441","author":"Mullen","year":"2012","journal-title":"Biochem. J."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"227","DOI":"10.1016\/j.theriogenology.2017.09.036","article-title":"De novo Synthesis of Sphingolipids Is Essential for Decidualization in Mice","volume":"106","author":"Ding","year":"2018","journal-title":"Theriogenology"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1016\/j.cellsig.2007.12.006","article-title":"The Sphingolipid Salvage Pathway in Ceramide Metabolism and Signaling","volume":"20","author":"Kitatani","year":"2008","journal-title":"Cell. Signal."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1093\/cvr\/cvp030","article-title":"Sphingomyelinases: Their Regulation and Roles in Cardiovascular Pathophysiology","volume":"82","author":"Pavoine","year":"2009","journal-title":"Cardiovasc. Res."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Shin, K.-O., Mihara, H., Ishida, K., Uchida, Y., and Park, K. (2022). Exogenous Ceramide Serves as a Precursor to Endogenous Ceramide Synthesis and as a Modulator of Keratinocyte Differentiation. Cells, 11.","DOI":"10.3390\/cells11111742"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Baker, P., Huang, C., Radi, R., Moll, S.B., Jules, E., and Arbiser, J.L. (2023). Skin Barrier Function: The Interplay of Physical, Chemical, and Immunologic Properties. Cells, 12.","DOI":"10.3390\/cells12232745"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"199","DOI":"10.4142\/jvs.2013.14.2.199","article-title":"Clinical Use of a Ceramide-Based Moisturizer for Treating Dogs with Atopic Dermatitis","volume":"14","author":"Jung","year":"2013","journal-title":"J. Vet. Sci."},{"key":"ref_28","first-page":"110","article-title":"Investigations on the Effects of a Topical Ceramides-Containing Emulsion (Allerderm Spot on) on Clinical Signs and Skin Barrier Function in Dogs with Topic Dermatitis: A Double-Blinded, Randomized, Controlled Study","volume":"11","author":"Marsella","year":"2013","journal-title":"Int. J. Appl. Res. Vet. Med."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"665","DOI":"10.1111\/j.1365-2230.2011.04313.x","article-title":"The Lipid Alterations in the Stratum Corneum of Dogs with Atopic Dermatitis are Alleviated by Topical Application of a Sphingolipid-Containing Emulsion: Skin Lipids of Dogs with AD Treated with Sphingolipids","volume":"37","author":"Popa","year":"2012","journal-title":"Clin. Exp. Dermatol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"812","DOI":"10.1111\/j.1365-4632.2008.03687.x","article-title":"Ceramide 1 and Ceramide 3 Act Synergistically on Skin Hydration and the Transepidermal Water Loss of Sodium Lauryl Sulfate-irritated Skin","volume":"47","author":"Huang","year":"2008","journal-title":"Int. J. Dermatol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"100400","DOI":"10.1016\/j.jlr.2023.100400","article-title":"Effect of Sphingosine and Phytosphingosine Ceramide Ratio on Lipid Arrangement and Barrier Function in Skin Lipid Models","volume":"64","author":"Rousel","year":"2023","journal-title":"J. Lipid Res."}],"container-title":["Animals"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/14\/2148\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T14:19:05Z","timestamp":1783693145000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2615\/16\/14\/2148"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,10]]},"references-count":31,"journal-issue":{"issue":"14","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["ani16142148"],"URL":"https:\/\/doi.org\/10.3390\/ani16142148","relation":{},"ISSN":["2076-2615"],"issn-type":[{"value":"2076-2615","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,10]]}}}