{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T10:16:56Z","timestamp":1780481816207,"version":"3.54.1"},"reference-count":48,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2026,5,31]],"date-time":"2026-05-31T00:00:00Z","timestamp":1780185600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Provincial Secretariat for Higher Education and Science, Autonomous Province of Vojvodina","award":["003877177 2025 09418 003 000 000 001"],"award-info":[{"award-number":["003877177 2025 09418 003 000 000 001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Background: Caffeine, although widely used in dermatological and cosmetic products, exhibits limited permeability through the stratum corneum, highlighting the need for strategies for optimizing delivery. The aim of this study was in vitro investigation of the effects of probiotic bacterial lysates and submicellar concentrations of bile acids on caffeine permeation, with a particular focus on permeation kinetics. Methods: Caffeine permeability was evaluated using the Skin Parallel Artificial Membrane Permeability Assay (Skin-PAMPA). Donor and acceptor concentrations were quantified by HPLC at predefined time points (1, 2, 4, 6, and 12 h), followed by calculation of apparent permeability coefficients, cumulative permeation profiles, and interval permeation rates in systems containing probiotic lysates and submicellar concentrations of cholic acid (CA) or deoxycholic acid (DCA). Results: Probiotic lysates significantly reduced caffeine permeability (0.98 \u00b1 0.02 \u00d7 10\u22126 vs. 1.57 \u00b1 0.14 \u00d7 10\u22126 cm\/s in the control group) and modified transport kinetics resulting in lower early-phase interval permeation rates and reduced cumulative permeation. Conversely, bile acids increased the apparent permeability of caffeine, with the highest value observed in the DCA group (2.30 \u00b1 0.08 \u00d7 10\u22126 cm\/s). Conclusions: Overall, probiotic lysates and bile acids modulated caffeine permeation across the Skin-PAMPA membrane primarily by reshaping permeation kinetics rather than simply changing overall permeability. Their combined effects may provide a basis for designing topical formulations with tailored permeation profiles.<\/jats:p>","DOI":"10.3390\/pharmaceutics18060688","type":"journal-article","created":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T09:33:14Z","timestamp":1780479194000},"page":"688","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Modulation of Caffeine Permeation Kinetics in a Skin-PAMPA Model by Probiotic Lysates and Bile Acids"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7785-0666","authenticated-orcid":false,"given":"Maja","family":"\u0110ani\u0107","sequence":"first","affiliation":[{"name":"Department of Pharmacology, Toxicology and Clinical Pharmacology, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalija","family":"Dedi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7186-3212","authenticated-orcid":false,"given":"Dragana","family":"Zaklan","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Slavica","family":"Lazarevi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacology, Toxicology and Clinical Pharmacology, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0708-3182","authenticated-orcid":false,"given":"Bojan","family":"Stanimirov","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2804-0624","authenticated-orcid":false,"given":"Momir","family":"Mikov","sequence":"additional","affiliation":[{"name":"Centre for Biomedical Research, Faculty of Medicine, University of Banja Luka, 78000 Banja Luka, Republic of Srpska, Bosnia and Herzegovina"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9444-5830","authenticated-orcid":false,"given":"Neboj\u0161a","family":"Pavlovi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pharmacy, Faculty of Medicine, University of Novi Sad, 21000 Novi Sad, Serbia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Dubashynskaya, N.V., and Skorik, Y.A. (2022). Patches as Polymeric Systems for Improved Delivery of Topical Corticosteroids: Advances and Future Perspectives. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232112980"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"103876","DOI":"10.1016\/j.xphs.2025.103876","article-title":"Sustained release of betamethasone from solid lipid nanoparticles loaded polymeric hydrogels with natural emollient: One step closer to effective topical therapy for atopic dermatitis","volume":"114","author":"Athavale","year":"2025","journal-title":"J. Pharm. Sci."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1468-2494.2008.00467.x","article-title":"Formulation effects of topical emulsions on transdermal and dermal delivery","volume":"31","author":"Otto","year":"2009","journal-title":"Int. J. Cosmet. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2698","DOI":"10.2174\/1381612821666150428124053","article-title":"Overcoming the skin permeation barrier: Challenges and opportunities","volume":"21","author":"Vitorino","year":"2015","journal-title":"Curr. Pharm. Des."},{"key":"ref_5","first-page":"291","article-title":"Radioprotective and antioxidant action of caffeine: Mechanistic considerations","volume":"34","author":"Devasagayam","year":"1996","journal-title":"Indian J. Exp. Biol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Rosado, C., Tokunaga, V.K., Sauce, R., de Oliveira, C.A., Sarruf, F.D., Parise-Filho, R., Maur\u00edcio, E., de Almeida, T.S., Velasco, M.V.R., and Baby, A.R. (2019). Another Reason for Using Caffeine in Dermocosmetics: Sunscreen Adjuvant. Front. Physiol., 10.","DOI":"10.3389\/fphys.2019.00519"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1208\/s12249-024-02943-2","article-title":"New Insight for Enhanced Topical Targeting of Caffeine for Effective Cellulite Treatment: In Vitro Characterization, Permeation Studies, and Histological Evaluation in Rats","volume":"25","author":"Fouad","year":"2024","journal-title":"AAPS PharmSciTech"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1159\/000343174","article-title":"Caffeine\u2019s Mechanisms of Action and Its Cosmetic Use","volume":"26","author":"Herman","year":"2013","journal-title":"Ski. Pharmacol. Physiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"874","DOI":"10.1002\/jps.10076","article-title":"Thermodynamic origin of the solubility profile of drugs showing one or two maxima against the polarity of aqueous and nonaqueous mixtures: Niflumic acid and caffeine","volume":"91","author":"Bustamante","year":"2002","journal-title":"J. Pharm. Sci."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"10957","DOI":"10.1021\/jp2021352","article-title":"Molecular dynamics simulation studies of caffeine aggregation in aqueous solution","volume":"115","author":"Tavagnacco","year":"2011","journal-title":"J. Phys. Chem. B"},{"key":"ref_11","first-page":"512","article-title":"Development and characterization of topical liposomal gel formulation for anti-cellulite activity","volume":"5","author":"Vyas","year":"2013","journal-title":"Int. J. Pharm. Pharm. Sci."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"e18236","DOI":"10.1590\/s2175-97902019000218236","article-title":"Nanoemulsion containing caffeine for cellulite treatment: Characterization and in vitro evaluation","volume":"55","author":"Freire","year":"2019","journal-title":"Braz. J. Pharm. Sci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"36","DOI":"10.3109\/10717544.2014.903011","article-title":"Design of solid lipid nanoparticles for caffeine topical administration","volume":"23","author":"Puglia","year":"2016","journal-title":"Drug Deliv."},{"key":"ref_14","first-page":"198","article-title":"The Characterization and Effectiveness Penetrasion of Caffeine Trapped and Coated Chitosan Nanoparticles as Anti-Cellulite","volume":"1","author":"Suptijah","year":"2015","journal-title":"J. Nanosci. Nanotechnol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"131","DOI":"10.1016\/0378-5173(94)90093-0","article-title":"Modulation of caffeine skin delivery by carrier design: Liposomes versus permeation enhancers","volume":"103","author":"Dayan","year":"1994","journal-title":"Int. J. Pharm."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Pavlovi\u0107, N., Bogi\u0107evi\u0107, I.A., Zaklan, D., \u0110ani\u0107, M., Golo\u010dorbin-Kon, S., Al-Salami, H., and Mikov, M. (2022). Influence of Bile Acids in Hydrogel Pharmaceutical Formulations on Dissolution Rate and Permeation of Clindamycin Hydrochloride. Gels, 8.","DOI":"10.3390\/gels8010035"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.colsurfb.2007.05.008","article-title":"Determination of critical micellar concentrations of cholic acid and its keto derivatives","volume":"59","author":"Posa","year":"2007","journal-title":"Colloids Surf. B Biointerfaces"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Pavlovi\u0107, N., Golo\u010dorbin-Kon, S., \u00d0ani\u0107, M., Stanimirov, B., Al-Salami, H., Stankov, K., and Mikov, M. (2018). Bile Acids and Their Derivatives as Potential Modifiers of Drug Release and Pharmacokinetic Profiles. Front. Pharmacol., 9.","DOI":"10.3389\/fphar.2018.01283"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"35660","DOI":"10.1074\/jbc.M113.519116","article-title":"Bile acids modulate signaling by functional perturbation of plasma membrane domains","volume":"288","author":"Zhou","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"\u0110ani\u0107, M., Pavlovi\u0107, N., Dedi\u0107, N., Zaklan, D., Lazarevi\u0107, S., Stanimirov, B., and Mikov, M. (2025). Influence of probiotics and deoxycholate on azathioprine transport in the PAMPA model: Insights into pharmacomicrobiomics and interindividual variability in drug response. Front. Pharmacol., 16.","DOI":"10.3389\/fphar.2025.1608110"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Dou, J., Feng, N., Guo, F., Chen, Z., Liang, J., Wang, T., Guo, X., and Xu, Z. (2023). Applications of Probiotic Constituents in Cosmetics. Molecules, 28.","DOI":"10.3390\/molecules28196765"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"34246","DOI":"10.1038\/s41598-025-16273-1","article-title":"Skin microbiome friendly topical formulations containing probiotic\u2014Loaded alginate microspheres: In vitro studies","volume":"15","author":"Michalczyk","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"16879","DOI":"10.1038\/s41598-023-43336-y","article-title":"Effects of a lotion containing probiotic ferment lysate as the main functional ingredient on enhancing skin barrier: A randomized, self-control study","volume":"13","author":"Cui","year":"2023","journal-title":"Sci. Rep."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Jung, Y.O., Jeong, H., Cho, Y., Lee, E.O., Jang, H.W., Kim, J., Nam, K., and Lim, K.M. (2019). Lysates of a Probiotic, Lactobacillus rhamnosus, Can Improve Skin Barrier Function in a Reconstructed Human Epidermis Model. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20174289"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Cui, H., Guo, C., Wang, Q., Feng, C., and Duan, Z. (2022). A pilot study on the efficacy of topical lotion containing anti-acne postbiotic in subjects with mild-to-moderate acne. Front. Med., 9.","DOI":"10.3389\/fmed.2022.1064460"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"16","DOI":"10.19080\/JOJDC.2018.02.555580","article-title":"Dead or Alive? A New Dilemma of Probiotic Skin Care for Healthier Skin","volume":"2","author":"Khmaladze","year":"2019","journal-title":"JOJ Dermatol. Cosmet."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"3427","DOI":"10.1111\/jocd.15831","article-title":"The pivotal role of Bifida Ferment Lysate on reinforcing the skin barrier function and maintaining homeostasis of skin defenses in vitro","volume":"22","author":"Wang","year":"2023","journal-title":"J. Cosmet. Dermatol."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Sink\u00f3, B., B\u00e1rdos, V., Vesztergombi, D., K\u00e1d\u00e1r, S., Malcsiner, P., Moustie, A., Jouy, C., Tak\u00e1cs-Nov\u00e1k, K., and Gr\u00e9goire, S. (2021). Use of an In Vitro Skin Parallel Artificial Membrane Assay (Skin-PAMPA) as a Screening Tool to Compare Transdermal Permeability of Model Compound 4-Phenylethyl-Resorcinol Dissolved in Different Solvents. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13111758"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3948","DOI":"10.1021\/jm060230+","article-title":"Parallel artificial membrane permeability assay: A new membrane for the fast prediction of passive human skin permeability","volume":"49","author":"Ottaviani","year":"2006","journal-title":"J. Med. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Ricci, A., Levante, A., Cirlini, M., Calani, L., Bernini, V., Del Rio, D., Galaverna, G., Neviani, E., and Lazzi, C. (2018). The Influence of Viable Cells and Cell-Free Extracts of Lactobacillus casei on Volatile Compounds and Polyphenolic Profile of Elderberry Juice. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.02784"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"219","DOI":"10.1007\/s13318-024-00878-4","article-title":"Influence of Bile Acids on Clindamycin Hydrochloride Skin Permeability: In Vitro and In Silico Preliminary Study","volume":"49","author":"Zaklan","year":"2024","journal-title":"Eur. J. Drug Metab. Pharmacokinet."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"291","DOI":"10.4236\/gsc.2023.134016","article-title":"Validation of a Method for the Measurement of Caffeine in Water by HPLC-UV","volume":"13","author":"Shehata","year":"2023","journal-title":"Green Sustain. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Wang, S.-S., Wang, C.-C., and Tung, C.-W. (2022). SkinSensPred as a Promising in Silico Tool for Integrated Testing Strategy on Skin Sensitization. Int. J. Environ. Res. Public Health, 19.","DOI":"10.3390\/ijerph191912856"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"105088","DOI":"10.1016\/j.isci.2022.105088","article-title":"Diffusion of small molecule drugs is affected by surface interactions and crowder proteins","volume":"25","author":"Dey","year":"2022","journal-title":"iScience"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3221","DOI":"10.1007\/s11033-020-05361-2","article-title":"Biological barriers, and the influence of protein binding on the passage of drugs across them","volume":"47","author":"Wanat","year":"2020","journal-title":"Mol. Biol. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1146\/annurev.biophys.37.032807.125817","article-title":"Macromolecular crowding and confinement: Biochemical, biophysical, and potential physiological consequences","volume":"37","author":"Zhou","year":"2008","journal-title":"Annu. Rev. Biophys."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1016\/j.ijpharm.2011.09.014","article-title":"Investigation of microemulsion microstructures and their relationship to transdermal permeation of model drugs: Ketoprofen, lidocaine, and caffeine","volume":"421","author":"Zhang","year":"2011","journal-title":"Int. J. Pharm."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1016\/S0168-3659(01)00248-6","article-title":"On the importance and mechanisms of burst release in matrix-controlled drug delivery systems","volume":"73","author":"Huang","year":"2001","journal-title":"J. Control. Release"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"3867","DOI":"10.1021\/bm501084u","article-title":"Affinity-based drug delivery systems for tissue repair and regeneration","volume":"15","author":"Vulic","year":"2014","journal-title":"Biomacromolecules"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Adepu, S., and Ramakrishna, S. (2021). Controlled Drug Delivery Systems: Current Status and Future Directions. Molecules, 26.","DOI":"10.3390\/molecules26195905"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1159\/000497225","article-title":"Permeation Mechanism of Caffeine and Naproxen through in vitro Human Epidermis: Effect of Vehicles and Penetration Enhancers","volume":"32","author":"Abd","year":"2019","journal-title":"Ski. Pharmacol. Physiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"101333","DOI":"10.1016\/j.jddst.2019.101333","article-title":"Effect of the aggregation state of bile salts on their transdermal absorption enhancing properties","volume":"54","year":"2019","journal-title":"J. Drug Deliv. Sci. Technol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"14451","DOI":"10.3390\/molecules200814451","article-title":"Absorption-Enhancing Effects of Bile Salts","volume":"20","author":"Moghimipour","year":"2015","journal-title":"Molecules"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"13461","DOI":"10.1021\/la203028s","article-title":"Effect of Submicellar Concentrations of Conjugated and Unconjugated Bile Salts on the Lipid Bilayer Membrane","volume":"27","author":"Mohapatra","year":"2011","journal-title":"Langmuir"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1159\/000441040","article-title":"A Comparison of the Penetration and Permeation of Caffeine into and Through Human Epidermis After Application in Various Vesicle Formulations","volume":"29","author":"Abd","year":"2016","journal-title":"Ski. Pharmacol. Physiol."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.ijpharm.2015.05.050","article-title":"Topical and transdermal delivery of caffeine","volume":"490","author":"Luo","year":"2015","journal-title":"Int. J. Pharm."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"S113","DOI":"10.4103\/2229-5178.146181","article-title":"Caffeine as a cause of urticaria-angioedema","volume":"5","author":"Tognetti","year":"2014","journal-title":"Indian Dermatol. Online J."},{"key":"ref_48","first-page":"163","article-title":"Skin models for the testing of transdermal drugs","volume":"8","author":"Abd","year":"2016","journal-title":"Clin. Pharmacol."}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/6\/688\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T09:42:30Z","timestamp":1780479750000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/18\/6\/688"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,31]]},"references-count":48,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6]]}},"alternative-id":["pharmaceutics18060688"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics18060688","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,31]]}}}