New photoactive nanofiber materials based on an aminolyzed polycaprolactone membrane with demonstrated cytocompatibility were developed. Two photoactive compounds, the photosensitizer Rose Bengal and the nitric oxide photodonor 4-nitro-3-(trifluoromethyl)aniline, were covalently bonded to the nanofiber surface, with or without a glutaraldehyde linker. The surface functionalization was confirmed via X-ray photoelectron spectroscopy, UV-vis absorption, and steady-state and time-resolved luminescence spectroscopy. Upon excitation with green or blue light, these materials efficiently generate antibacterial species, including singlet oxygen, with a slight contribution of hydrogen peroxide and nitric oxide. A potent light-induced antibacterial effect was demonstrated against Escherichia coli. Furthermore, the functionalized photoactive membranes, especially those with a glutaraldehyde linker and photosterilized by light, not only excluded the material toxicity but also demonstrated improved cell adhesion and proliferation when tested with adipose tissue-derived stem cells. These materials, which offer a unique combination of light-controlled surface sterilization and high cellular compatibility, are promising for advanced tissue engineering applications.

Faculty of Sciences Charles University Hlavova 2030 Prague 2 128 43 Czech Republic

Institute of Macromolecular Chemistry Academy of Sciences of the Czech Republic Heyrovského nám 2 Prague 6 162 00 Czech Republic

Institute of Physiology of the Czech Academy of Sciences Vídeňská 1083 Prague 4 142 00 Czech Republic

J Heyrovský Institute of Physical Chemistry v v i Academy of Sciences of the Czech Republic Dolejškova 3 Prague 8 182 23 Czech Republic

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Santoro M., Shah S. R., Walker J. L., Mikos A. G.. Poly­(lactic acid) nanofibrous scaffolds for tissue engineering. Adv. Drug Delivery Rev. 2016;107:206–212. doi: 10.1016/j.addr.2016.04.019. PubMed DOI PMC

Rahmati M., Mills D. K., Urbanska A. M., Saeb M. R., Venugopal J. R., Ramakrishna S., Mozafari M.. Electrospinning for tissue engineering applications. Prog. Mater. Sci. 2021;117:100721. doi: 10.1016/j.pmatsci.2020.100721. DOI

Ambekar R. S., Kandasubramanian B.. Advancements in nanofibers for wound dressing: A review. Eur. Polym. J. 2019;117:304–336. doi: 10.1016/j.eurpolymj.2019.05.020. DOI

Lu T., Cui J., Qu Q., Wang Y., Zhang J., Xiong R., Ma W., Huang C.. Multistructured Electrospun Nanofibers for Air Filtration: A Review. ACS Appl. Mater. Interfaces. 2021;13(20):23293–23313. doi: 10.1021/acsami.1c06520. PubMed DOI

Kalantari K., Afifi A. M., Jahangirian H., Webster T. J.. Biomedical applications of chitosan electrospun nanofibers as a green polymer – Review. Carbohydr. Polym. 2019;207:588–600. doi: 10.1016/j.carbpol.2018.12.011. PubMed DOI

Kenry, Lim C. T.. Nanofiber technology: Current status and emerging developments. Prog. Polym. Sci. 2017;70:1–17. doi: 10.1016/j.progpolymsci.2017.03.002. DOI

Henke P., Lang K., Kubát P., Sýkora J., Šlouf M., Mosinger J.. Polystyrene Nanofiber Materials Modified with an Externally Bound Porphyrin Photosensitizer. ACS Appl. Mater. Interfaces. 2013;5(9):3776–3783. doi: 10.1021/am4004057. PubMed DOI

Asghari F., Samiei M., Adibkia K., Akbarzadeh A., Davaran S.. Biodegradable and biocompatible polymers for tissue engineering application: A review. Artif. Cells, Nanomed., Biotechnol. 2017;45(2):185–192. doi: 10.3109/21691401.2016.1146731. PubMed DOI

Khorshidi S., Solouk A., Mirzadeh H., Mazinani S., Lagaron J. M., Sharifi S., Ramakrishna S.. A review of key challenges of electrospun scaffolds for tissue-engineering applications. J. Tissue Eng. Regener. Med. 2016;10(9):715–738. doi: 10.1002/term.1978. PubMed DOI

Chen S., Liu B., Carlson M. A., Gombart A. F., Reilly D. A., Xie J.. Recent Advances in Electrospun Nanofibers for Wound Healing. Nanomedicine. 2017;12(11):1335–1352. doi: 10.2217/nnm-2017-0017. PubMed DOI PMC

Liu S., Yu J.-M., Gan Y.-C., Qiu X.-Z., Gao Z.-C., Wang H., Chen S.-X., Xiong Y., Liu G.-H., Lin S.-E.. et al. Biomimetic Natural Biomaterials for Tissue Engineering and Regenerative Medicine: New Biosynthesis Methods, Recent Advances, and Emerging Applications. Mil Med. Res. 2023;10(1):16. doi: 10.1186/s40779-023-00448-w. PubMed DOI PMC

Dai Z., Ronholm J., Tian Y., Sethi B., Cao X.. Sterilization techniques for biodegradable scaffolds in tissue engineering applications. J. Tissue Eng. 2016;7:2041731416648810. doi: 10.1177/2041731416648810. PubMed DOI PMC

Liu S., Qin S., He M., Zhou D., Qin Q., Wang H.. Current applications of poly­(lactic acid) composites in tissue engineering and drug delivery. Composites, Part B. 2020;199:108238. doi: 10.1016/j.compositesb.2020.108238. DOI

Pawar R., Pathan A., Nagaraj S., Kapare H., Giram P., Wavhale R.. Polycaprolactone and its derivatives for drug delivery. Polym. Adv. Technol. 2023;34(10):3296–3316. doi: 10.1002/pat.6140. DOI

Yaseri R., Fadaie M., Mirzaei E., Samadian H., Ebrahiminezhad A.. Surface modification of polycaprolactone nanofibers through hydrolysis and aminolysis: A comparative study on structural characteristics, mechanical properties, and cellular performance. Sci. Rep. 2023;13(1):9434. doi: 10.1038/s41598-023-36563-w. PubMed DOI PMC

Ehtesabi H., Massah F.. Improvement of hydrophilicity and cell attachment of polycaprolactone scaffolds using green synthesized carbon dots. Mater. Today. Sustain. 2021;13:100075. doi: 10.1016/j.mtsust.2021.100075. DOI

Jeznach O., Kołbuk D., Marzec M., Bernasik A., Sajkiewicz P.. Aminolysis as a surface functionalization method of aliphatic polyester nonwovens: impact on material properties and biological response. RSC Adv. 2022;12(18):11303–11317. doi: 10.1039/D2RA00542E. PubMed DOI PMC

Yuan S., Xiong G., Wang X., Zhang S., Choong C.. Surface modification of polycaprolactone substrates using collagen-conjugated poly­(methacrylic acid) brushes for the regulation of cell proliferation and endothelialisation. J. Mater. Chem. 2012;22(26):13039–13049. doi: 10.1039/c2jm31213a. DOI

Zhu Y., Mao Z., Shi H., Gao C.. In-depth study on aminolysis of poly­(ε-caprolactone): Back to the fundamentals. Sci. China: Chem. 2012;55(11):2419–2427. doi: 10.1007/s11426-012-4540-y. DOI

Grabska-Zielińska S.. Cross-Linking Agents in Three-Component Materials Dedicated to Biomedical Applications: A Review. Polymers. 2024;16(18):2679. doi: 10.3390/polym16182679. PubMed DOI PMC

Rediguieri C. F., Sassonia R. C., Dua K., Kikuchi I. S., de Jesus Andreoli Pinto T.. Impact of sterilization methods on electrospun scaffolds for tissue engineering. Eur. Polym. J. 2016;82:181–195. doi: 10.1016/j.eurpolymj.2016.07.016. DOI

Horakova J., Klicova M., Erben J., Klapstova A., Novotny V., Behalek L., Chvojka J.. Impact of Various Sterilization and Disinfection Techniques on Electrospun Poly-ε-caprolactone. ACS Omega. 2020;5(15):8885–8892. doi: 10.1021/acsomega.0c00503. PubMed DOI PMC

Valente T. A. M., Silva D. M., Gomes P. S., Fernandes M. H., Santos J. D., Sencadas V.. Effect of Sterilization Methods on Electrospun Poly­(lactic acid) (PLA) Fiber Alignment for Biomedical Applications. ACS Appl. Mater. Interfaces. 2016;8(5):3241–3249. doi: 10.1021/acsami.5b10869. PubMed DOI

Wade M. B., Rodenberg E., Patel U., Shah B., Becker M. L.. Influence of Sterilization Technologies on Electrospun Poly­(ester urea)­s for Soft Tissue Repair. Biomacromolecules. 2016;17(10):3363–3374. doi: 10.1021/acs.biomac.6b01158. PubMed DOI

Elashnikov R., Rimpelová S., Vosmanská V., Kolská Z., Kolářová K., Lyutakov O., Švorčík V.. Effect of sterilization methods on electrospun cellulose acetate butyrate nanofibers for SH-SY5Y cultivation. React. Funct. Polym. 2019;143:104339. doi: 10.1016/j.reactfunctpolym.2019.104339. DOI

Liška V., Willimetz R., Kubát P., Křtěnová P., Gyepes R., Mosinger J.. Synergistic photogeneration of nitric oxide and singlet oxygen by nanofiber membranes via blue and/or red-light irradiation: Strong antibacterial action. J. Photochem. Photobiol., B. 2024;255:112906. doi: 10.1016/j.jphotobiol.2024.112906. PubMed DOI

Henke P., Kozak H., Artemenko A., Kubát P., Forstová J., Mosinger J.. Superhydrophilic Polystyrene Nanofiber Materials Generating O2­(1Δg): Postprocessing Surface Modifications toward Efficient Antibacterial Effect. ACS Appl. Mater. Interfaces. 2014;6(15):13007–13014. doi: 10.1021/am502917w. PubMed DOI

Mosinger J., Jirsák O., Kubát P., Lang K., Mosinger B.. Bactericidal nanofabrics based on photoproduction of singlet oxygen. J. Mater. Chem. 2007;17(2):164–166. doi: 10.1039/B614617A. DOI

Mosinger J., Lang K., Kubát P., Sýkora J., Hof M., Plíštil L., Mosinger B.. Photofunctional Polyurethane Nanofabrics Doped by Zinc Tetraphenylporphyrin and Zinc Phthalocyanine Photosensitizers. J. Fluoresc. 2009;19(4):705–713. doi: 10.1007/s10895-009-0464-0. PubMed DOI

Jesenská S., Plíštil L., Kubát P., Lang K., Brožová L., Popelka Š., Szatmáry L., Mosinger J.. Antibacterial nanofiber materials activated by light. J. Biomed. Mater. Res., Part A. 2011;99A(4):676–683. doi: 10.1002/jbm.a.33218. PubMed DOI

Arenbergerova M., Arenberger P., Bednar M., Kubat P., Mosinger J.. Light-activated nanofibre textiles exert antibacterial effects in the setting of chronic wound healing. Exp. Dermatol. 2012;21(8):619–624. doi: 10.1111/j.1600-0625.2012.01536.x. PubMed DOI

Lhotáková Y., Plíštil L., Morávková A., Kubát P., Lang K., Forstová J., Mosinger J.. Virucidal Nanofiber Textiles Based on Photosensitized Production of Singlet Oxygen. PLoS One. 2012;7(11):e49226. doi: 10.1371/journal.pone.0049226. PubMed DOI PMC

Mosinger J., Lang K., Plíštil L., Jesenská S., Hostomský J., Zelinger Z., Kubát P.. Fluorescent Polyurethane Nanofabrics: A Source of Singlet Oxygen and Oxygen Sensing. Langmuir. 2010;26(12):10050–10056. doi: 10.1021/la1001607. PubMed DOI

Dolanský J., Henke P., Kubát P., Fraix A., Sortino S., Mosinger J.. Polystyrene Nanofiber Materials for Visible-Light-Driven Dual Antibacterial Action via Simultaneous Photogeneration of NO and O2­(1Δg) ACS Appl. Mater. Interfaces. 2015;7(41):22980–22989. doi: 10.1021/acsami.5b06233. PubMed DOI

Wang T.-Y., Zhu X.-Y., Wu F.-G.. Antibacterial gas therapy: Strategies, advances, and prospects. Bioactive Mater. 2023;23:129–155. doi: 10.1016/j.bioactmat.2022.10.008. PubMed DOI PMC

Neckers D. C.. Rose Bengal. J. Photochem. Photobiol. A Chem. 1989;47(1):1–29. doi: 10.1016/1010-6030(89)85002-6. DOI

Caruso E. B., Petralia S., Conoci S., Giuffrida S., Sortino S.. Photodelivery of Nitric Oxide from Water-Soluble Platinum Nanoparticles. J. Am. Chem. Soc. 2007;129(3):480–481. doi: 10.1021/ja067568d. PubMed DOI

Sharma N., Dhyani A. K., Marepally S., Jose D. A.. Nanoscale lipid vesicles functionalized with a nitro-aniline derivative for photoinduced nitric oxide (NO) delivery. Nanoscale Adv. 2020;2(1):463–469. doi: 10.1039/C9NA00532C. PubMed DOI PMC

Noel S., Liberelle B., Robitaille L., De Crescenzo G.. Quantification of Primary Amine Groups Available for Subsequent Biofunctionalization of Polymer Surfaces. Bioconjugate Chem. 2011;22(8):1690–1699. doi: 10.1021/bc200259c. PubMed DOI

Mosinger J., Mosinger B.. Photodynamic sensitizers assay: Rapid and sensitive iodometric measurement. Experientia. 1995;51(2):106–109. doi: 10.1007/BF01929349. PubMed DOI

Corbett J. T.. The scopoletin assay for hydrogen peroxide A review and a better method. J. Biochem Biophys. Met. 1989;18(4):297–307. doi: 10.1016/0165-022X(89)90039-0. PubMed DOI

Fong H., Chun I., Reneker D. H.. Beaded nanofibers formed during electrospinning. Polymer. 1999;40(16):4585–4592. doi: 10.1016/S0032-3861(99)00068-3. DOI

Suchánek J., Henke P., Mosinger J., Zelinger Z., Kubát P.. Effect of Temperature on Photophysical Properties of Polymeric Nanofiber Materials with Porphyrin Photosensitizers. J. Phys. Chem. B. 2014;118(23):6167–6174. doi: 10.1021/jp5029917. PubMed DOI

Bregnhøj M., Westberg M., Jensen F., Ogilby P. R.. Solvent-dependent singlet oxygen lifetimes: Temperature effects implicate tunneling and charge-transfer interactions. Phys. Chem. Chem. Phys. 2016;18(33):22946–22961. doi: 10.1039/C6CP01635A. PubMed DOI

Rong F., Tang Y., Wang T., Feng T., Song J., Li P., Huang W.. Nitric Oxide-Releasing Polymeric Materials for Antimicrobial Applications: A Review. Antioxidants. 2019;8(11):556. doi: 10.3390/antiox8110556. PubMed DOI PMC

Sun J., Zhang X., Broderick M., Fein H.. Measurement of Nitric Oxide Production in Biological Systems by Using Griess Reaction Assay. Sensors. 2003;3(8):276–284. doi: 10.3390/s30800276. DOI

Goh M. H., Connolly J. J., Chen A. F., Rabiner R. A., Lozano-Calderon S. A.. Antimicrobial effect of blue light on antibiotic-sensitive and drug-resistant Escherichia coli: A novel isotropic optical fibre. Access Microbiol. 2025;7(3):000967–v3. doi: 10.1099/acmi.0.000967.v3. PubMed DOI PMC

Filová E., Staňková L., Eckhardt A., Svobodová J., Musílková J., Pala J., Hadraba D., Brynda E., Koňařík M., Pirk J., Bačáková L.. Modification of Human Pericardium by Chemical Crosslinking. Physiol. Res. 2020;69(1):49–59. doi: 10.33549/physiolres.934335. PubMed DOI PMC

Musilkova J., Filova E., Pala J., Matejka R., Hadraba D., Vondrasek D., Kaplan O., Riedel T., Brynda E., Kucerova J.. et al. Human Decellularized and Crosslinked Pericardium Coated with Bioactive Molecular Assemblies. Biomed. Mater. 2019;15(1):015008. doi: 10.1088/1748-605X/ab52db. PubMed DOI