Dynamics of nanoparticles transport through cellular barriers relevant for toxicology and drug delivery systems

Project title: Dynamics of nanoparticles transport through cellular barriers relevant for toxicology and drug delivery systems

Project director: Assoc. Prof. Miruna-Silvia Stan

University of Bucharest/Faculty of Biology/Dep. of Biochemistry and Molecular Biology

Tel: +40761976161

E-mail: miruna.stan@bio.unibuc.ro

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Summary: As the full understanding of the mechanism of nanoparticles (NPs) transport across the biological barriers has not yet been achieved, re-evaluation of the in vitro models used to analyze the transport in correlation with the NPs characteristics is mandatory. The present project aims to perform a complex analysis of the biological effects and transport mechanism of NPs with different characteristics (chemical structures, sizes, surfaces) through several important biological barriers (skin, lung, intestinal and blood-brain barriers). The results will generate robust computational 3D models for predicting the behavior, performance and toxicity of NPs in vivo, and also for designing nano-devices with improved efficiency in this “online” era. Using commercially available in vitro barriers or cell culture models developed by the team members based on their expertise, the effects triggered during exposure to NPs will be analyzed. The implementation of the specific objectives will bring multiple elements of originality and innovation in the field of cell culture models, nanotoxicology, nanomedicine and bioinformatics, in which the project leader and team members have significant and representative scientific achievements. This project has an important social and economic impact, with applicable directions in drug delivery purposes, diagnostic imaging or other biomedical applications.

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Funding agency: UEFISCDI

Contract number: 81TE/2022

Project code: PN-III-P1-1.1-TE-2021-1375

Project acronym:  TRANS-NANO-BIO

Project duration: 24 months (12.05.2022 – 11.05.2024)

Total funding: 450000 RON

Project members: Conf. Dr. Miruna-Silvia STAN (project leader)

                               Conf. Dr. Maria MERNEA (postdoctoral researcher)

                               Lect. Dr. Sorina VOICU (postdoctoral researcher)

                               Dr. Cristina VOINEA (postdoctoral researcher)

                               Drd. Mirela SERBAN (PhD student)

                               Drd. Bogdan MIU (PhD student)

Project objective: The main objective of this research project is represented by the study of nanoparticles transport and toxicity across biological barriers using innovative, mechanistic-based strategies, that are of primary interest for nanotoxicology and for designing highly biocompatible and efficient next-generation therapy nano-devices.

Expected results for this project

– validation of currently available in vitro models to study the nanoparticles biological effects and transport,

– obtaining insights regarding the mechanism of nanoparticles translocation, and the correlation with their physicochemical characteristics,

– generation of predictive models for in vivo biological effects and performance of nanoparticles delivery systems,

– scientific peer-reviewed publications and results’ presentation as posters and talks at international conferences.

Results of Phase I – 2022

– poster at international conferences

• Voicu S.N., Voinea I.C., Stan M.S. “Assessment of the in vitro toxicity induced by rare earth-doped phosphor nanoparticles on human lung fibroblast cells”, NanoBioMat2022 winter edition, 24-26 November 2022, Online.
• Stan M.S., Voinea I.C., Voicu S.N. “In vitro evaluation of rare earth-doped phosphor nanoparticles to assess their anti-tumoral efficiency on lung cancer cells”, The 8st International Electronic Conference on Medicinal Chemistry, 1–30 November 2022. https://doi.org/10.3390/ECMC2022-13312

– research article published in ISI journal (Q1)

• Florea DA, Grumezescu V, Bîrcă AC, Vasile BȘ, Iosif A, Chircov C, Stan MS, Grumezescu AM, Andronescu E, Chifiriuc MC. Bioactive hydroxyapatite-magnesium phosphate coatings deposited by MAPLE for preventing infection and promoting orthopedic implants osteointegration. Materials 2022; 15(20), 7337. https://doi.org/10.3390/ma15207337

– review paper published in ISI journal (Q2)

• Miu BA, Dinischiotu A. New green approaches in nanoparticles synthesis: an overview. Molecules. 2022; 27(19):6472. https://doi.org/10.3390/molecules27196472

– participation at practical course

• EMBO Practical Course “Current methods in cell biology”, Heidelberg, Germany, 4 – 13 September 2022.

The summary of the results obtained

Results of Phase II – 2023:

– posters at international conferences

• Voicu S., Voinea I.C., Niculescu A., Mihaescu D., Grumezescu A., Stan M. “Characterizing the biological behavior of Fe₃O₄ nanoparticles conjugated with acridine orange using in vitro co-culture systems relevant to skin, lung and gut barrier models”, in Proceedings of the 4th International Electronic Conference on Applied Sciences, 27 October–10 November 2023, MDPI: Basel, Switzerland, Online. https://sciforum.net/paper/view/15411
• Adeniyi, B., Pieraccini, L., Cirstea, G., Vilcoci, S., Mernea, M., Stan M. “Evaluation of the anti-cancer activity of Goji leaves extracts against HT-29-MTX colon cancer cell line”, in Proceedings of the 4th International Electronic Conference on Foods, 15–30 October 2023, MDPI: Basel, Switzerland, Online. https://sciforum.net/paper/view/15016
• Dula A.G., Stan M.S. “Fluorescent PLGA nanoparticles: in vitro toxicity study using a co-culture of human keratinocytes and dermal fibroblasts”, NanoInnovation2023, 18-22 September 2023, Rome, Italy.
• Voicu S.N., Voinea I.C., Gosee C., Carteret C., Callewaert M., Moreau J., Chuburu F., Stan M.S. “Biocompatibility evaluation of chitosan fluorescent nanohydrogels using human EpiDerm^TM 3D model”. Toxicology Letters 384S1 (2023) S73-S300, p. S120-121. The 57th Congress of the European Societies of Toxicology (EUROTOX 2023), Ljubljana, Slovenia, 10 –13 September 2023.
• Voinea I.C., Voicu S.N., Mernea M., Miu B.A., Serban M.V., Stan M.S. “Prediction of respiratory toxicity induced by P25 Degussa nanoparticles using human EpiAirway™ 3D model”. Toxicology Letters 384S1 (2023) S73-S300, p. S172. The 57th Congress of the European Societies of Toxicology (EUROTOX 2023), Ljubljana, Slovenia, 10 –13 September 2023.
• Voinea I.C., Voicu S., Mernea M., Miu B., Serban M., Stan M. “Using EpiDermTM to evaluate the toxic effects of P25 Degussa nanoparticles on skin barrier”, in Proceedings of the 4th International Online Conference on Nanomaterials, 5–19 May 2023, MDPI: Basel, Switzerland, Online. https://sciforum.net/paper/view/14460

– research articles published in ISI journals (Q2)

• Burdusel A-C, Neacsu IA, Birca AC, Chircov C, Grumezescu A-M, Holban AM, Curutiu C, Ditu LM, Stan M, Andronescu E. Microwave-assisted hydrothermal treatment of multifunctional substituted hydroxyapatite with prospective applications in bone regeneration. Funct. Biomater. 2023, 14(7), 378. https://doi.org/10.3390/jfb14070378
• Marinescu G, Culita DC, Mocanu T, Mitran R-A, Petrescu S, Stan MS, Chifiriuc MC, Popa M. New nanostructured materials based on mesoporous silica loaded with Ru(II)/Ru(III) complexes with anticancer and antimicrobial properties. Pharmaceutics 2023, 15(5),1458. https://doi.org/10.3390/pharmaceutics15051458
• Miu BA, Voinea IC, Diamandescu L, Dinischiotu A. MRC-5 human lung fibroblasts alleviate the genotoxic effect of Fe-N co-doped titanium dioxide nanoparticles through an OGG1/2-dependent reparatory mechanism. International Journal of Molecular Sciences 2023, 24(7), 6401. https://doi.org/10.3390/ijms24076401

Scientific and Technical Report

Results of Phase III – 2024:

– posters at international conferences

• Mernea M., Voicu S., Voinea C., Stan M. “Simulation on the interaction between FITC functionalized gold nanoparticles and lipid bilayers”, in Proceedings of the 3rd International Electronic Conference on Biomolecules, 23–25 April 2024, MDPI: Basel, Switzerland.
• Stan M.S., Dungu C., Peligrad M.I., Daraban B.S., et al. “Crossing the pulmonary and dermal barriers by PLGA nanoparticles. An in vitro study with cell co-culture models”, FEBS Open Bio 14 (Suppl. S2) (2018), p. 456 (FEBS 2024 – 48th FEBS Congress, 29 June–3 July 2024, Milano, Italy).

– research articles published in ISI journals (Q1)

• Gosée C, Moreau J, Cadiou C, Callewaert M, Henoumont C, Larbanoix L, Molinari M, Voicu SN, et al. Multimodal nanogels combining ZW800-1 as an optical absorber and gadolinium chelates for multispectral optoacoustic tomography (MSOT) and magnetic resonance imaging (MRI). Chem. Front. 2023, 10, 6357-6368.
• Raduly FM, Raditoiu V, Raditoiu A, Nicolae CA, Grapin M, Stan MS, Voinea IC, et al. Half-Curcuminoids Encapsulated in Alginate–Glucosamine Hydrogel Matrices as Bioactive Delivery Systems. Gels 2024, 10(6), 376.
• Miu BA, Stan MS, Mernea M, Dinischiotu A, Voinea IC. Pure epigallocatechin-3-gallate-assisted green synthesis of highly stable titanium dioxide nanoparticles. Materials 2024, 17(2), 275.

Results obtained within this project:

The project has achieved significant results, notably in developing in vitro cellular models such as the intestinal barrier and blood-brain barrier. Advances in skin and lung models using human cell co-cultures have led to new 3D cellular systems for testing drug delivery capacities. The mechanism of nanoparticle translocation was highlighted, correlating their physicochemical characteristics to create predictive models for biological effects in vivo and delivery system performance. Both gold and PLGA nanoparticles demonstrated low cytotoxicity, making them biocompatible and suitable for drug delivery applications. Studies revealed these nanoparticles can traverse the blood-brain barrier model, the gold nanoparticles showing exceptional potential. Additionally, PLGA nanoparticles were internalized by lung cells within 24 hours, while gold nanoparticles showed increased transport rate across the intestinal barrier after 48 hours, suggesting their efficiency in developing gastrointestinal treatments. The data collected within this project enhances our understanding of cellular responses to different types of nanoparticles, offering safer and more effective nanomedicine strategies. This project has met all proposed activities, resulted in eight scientific articles (4 Q1 + 4 Q2; Total IF = 36.3, Total AIS = 5.822), ten conference participations, and established ongoing collaborations with external research groups, laying the groundwork for future research projects.

Short Report 2024