Összes szerző

Péter Beatrix

az alábbi absztraktok szerzői között szerepel:

Farkas Enikő
Controlling Live Cell Adhesion through Characterization of Biofunctionalized Surfaces using Label-Free Biosensors

Aug 30 - szerda

15:30 – 17:00

II. Poszterszekció

P41

Controlling Live Cell Adhesion through Characterization of Biofunctionalized Surfaces using Label-Free Biosensors

Eniko Farkas¹, Kinga Dóra Kovács^1,3, Beatrix Peter¹, Attila Bonyár², Sandor Kurunczi¹, Inna Szekacs¹, and Robert Horvath¹

¹ Nanobiosensorics Laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research, Budapest, Hungary

² Department of Electronics Technology, Faculty of Electrical Engineering and Informatics, Budapest University of Technology and Economics, Budapest, Hungary

Biomaterial coatings that possess cell-repellent or cell-adhesive properties have a significant interest in medical and biotechnological applications [1-4]. However, conventional approaches lack in-depth analysis and quantitative comparison of these coatings for regulating adhesion, particularly for bacterial cell adhesion. Label-free Optical Waveguide Lightmode Spectroscopy (OWLS) can offer a solution for the detailed analysis of biomaterial coatings. OWLS biosensors detect the optical properties of the adhesive surface using evanescent waves with a penetration depth of 100-150 nm [5-7]. This surface-sensitive technique enables a thorough evaluation of biomaterial coatings for regulating adhesion. Uniquely, OWLS enables the in situ measurement of both the coating process and subsequent cell adhesion.

The present study utilizes the OWLS method for in-depth characterization of biomaterial surfaces with regard to bacterial adhesion. Initially, adhesion blocking biomaterials, namely bovine serum albumin, I-block, PAcrAM-g-(PMOXA, NH2, Si), (PAcrAM-P), and PLL-g-PEG, with varying coating temperatures, were screened. PAcrAM-P exhibited the best blocking capability against bacterial concentrations up to 10⁷ cells/mL. Subsequently, different immobilization methods, such as Mix&Go (AnteoBind) films, protein A, avidin-biotin based surface chemistries, and simple physisorption, were employed to captureEscherichia coli specific antibodies. Bacterial cell adhesion was then tested on immobilized antibodies with various blocking agents. The OWLS analysis allowed for the determination of the parameters of the applied agents by considering the kinetic data of adhesion, the surface mass density, and the protein orientation. Based on the experimental results, surfaces were created and tested for controlling both bacterial and mammalian cell adhesion. [8]

Acknowledgment

This work was supported by the "Lendület" (HAS) research program, the National Research, Development and Innovation Office of Hungary ((ERC_HU, VEKOP 2.2.1-16, ELKH topic-fund, "Élvonal" KKP_19 and KH grants, PD 131543 and TKP2022-EGA-04 –INBIOM TKP Programs financed from the NRDI Fund). This work was also supported by 77 Elektronika Ltd. by their supplying of antibodies and reagents.

References

[1] Frutiger A, et. al. (2021) Chem Rev 121: 8095–8160.

[2] Rigo S, et. al. (2018) Adv Sci 5: 1700892.

[3] Castillo-Henríquez L, et. al. (2020) Sensors 20: 6926.

[4] D’Agata R, et. al. (2021) Polymers 13:1929.

[5] Vörös J, et. al. (2002) Biomaterials 23: 3699–3710.

[6] Tiefenthaler K, et. al. (1989) J Opt Soc Am. B 6: 209–220.

[7] Saftics A, et. al. (2021) Adv Colloid Interface Sci 294: 102431–102433.

[8] Farkas E, et. al. (2022) Biosensors 12: 56.

Kovács Dóra Kinga
Nanoinjection of fluorescent nanoparticles to single live cells by robotic fluidic force microscopy

Aug 29 - kedd

12:15 – 12:30

Modern biofizikai módszerek

E13

Nanoinjection of fluorescent nanoparticles to single live cells by robotic fluidic force microscopy

Kinga Dóra Kovács^1,*, Tamás Visnovitz^2,3,*, Tamás Gerecsei¹, Beatrix Peter¹, Sándor Kurunczi¹, Anna Koncz², Krisztina Németh², Dorina Lenzinger², Krisztina V. Vukman², Péter Lőrincz⁴, Inna Székács¹, Edit I. Buzás^2,5,6**, Robert Horvath^1,**

¹ Nanobiosensorics Laboratory, Centre of Energy Research, ELKH, Budapest, Hungary

² Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary

³ Department of Plant Physiology and Molecular Plant Biology, ELTE Eötvös Loránd University, Budapest, Hungary

⁴ Department of Anatomy, Cell and Developmental Biology, ELTE Eötvös Loránd University, Budapest, Hungary

⁵ HCEMM-SU Extracellular Vesicle Research Group, Budapest, Hungary

⁶ ELKH-SE Translational Extracellular Vesicle Research Group, Budapest, Hungary

*,** equal contributions / **corresponding authors

Direct injection of fluorescent nanoparticles into the cytoplasm of living cells can provide new insights into the intracellular fate of various different fluorescently labelled biologically active particles. Here we used fluorescent nanoparticles to prove the feasibility of nanoinjection into single live HeLa cells by using robotic fluidic force microscopy (FluidFM). This injection platform offers the advantage of high cell selectivity and efficiency. We confirmed the successful injection of both GFP encoding plasmids and GFP tagged fluorescent nanoparticles to the cells by confocal microscopy. We were able track the nanoparticles in the living cells for 20 hours. The injected nanoparticles were initially localized in concentrated spot-like regions within the cytoplasm. Later, they were transported towards the periphery of the cells. Based on our proof-of-principle data, the FluidFM platform is suitable for targeting single living cells by fluorescently labelled biologically active particles and may lead to information about the intracellular cargo delivery at a single-cell level.

Péter Beatrix
Nanoparticle uptake of living cells with digested glycocalyx

Aug 29 - kedd

15:30 – 17:00

I. Poszterszekció

P21

Nanoparticle uptake of living cells with digested glycocalyx

Beatrix Peter^a, Nicolett Kanyo^a, Kinga Dora Kovacs^a,b, Viktor Kovács^a, Inna Szekacs^a, Béla Pécz^c, Kinga Molnár^d,Hideyuki Nakanishi^e, Istvan Lagzi^,,f,g, Robert Horvath^a

^a Nanobiosensorics laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research, Konkoly-Thege út 29-33, H-1121 Budapest, Hungary

^b Department of Biological Physics, Eötvös University, Budapest, Hungary

^c Thin Films Laboratory, Institute of Technical Physics and Materials Science, Centre for Energy Research, Konkoly-Thege út 29-33, H-1120 Budapest, Hungary

^d Department of Anatomy, Cell and Developmental Biology, ELTE, Eötvös Loránd University, Pázmány Péter stny. 1/C, Budapest, H-1117, Hungary

^e Department of Macromolecular Science and Engineering, Graduate School of Science and Technology, Kyoto Institute of Technology, Matsugasaki, Kyoto 606-8585, Japan

^f Department of Physics, Institute of Physics, Budapest University of Technology and Economics, Műegyetem rkp. 3, Budapest H-1111, Hungary

^g ELKH BME Condensed Matter Research Group, Műegyetem rkp. 3, Budapest H-1111, Hungary

In biomedical imaging and targeted drug delivery, functionalized nanoparticles are widely used due to their penetration into living cells. The glycocalyx is a surface sugar layer of the cells, which presumably plays an essential role in any uptake process. However, its exact function in nanoparticle uptake is still uncovered. We in situ monitored the penetration of positively charged gold nanoparticles into adhered cancer cells with or without preliminary glycocalyx digestion. During the experiments, the components of glycocalyx of HeLa cells were digested by chondroitinase ABC enzyme. The measurements were performed by applying a high-throughput label-free resonant waveguide grating biosensor. The positively charged gold nanoparticles were used with different sizes (S, M, L). Negatively charged citrate-capped tannic acid nanoparticles, and other types of glycocalyx digesting enzymes were also applied in control experiments. The biosensor data confirmed the cellular uptake of the functionalized nanoparticles with an active process, which was verified by transmission electron microscopy [1,2]. Based on the findings we conclude that the components of gylcocalyx control the uptake process in size- and charge-dependent manner, and the possible roles of various glycocalyx components were highlighted.

Acknowledgements

This work was supported by the National Research, Development, and Innovation Office (Grant Numbers: PD 131543 for B.P., ELKH topic-fund, "Élvonal" KKP_19 TKP2022-EGA-04 grants).

References

[1] B. Peter, N. Kanyo, K. D. Kovacs, V. Kovács, I. Szekacs, B. Pécz, K. Molnár, H. Nakanishi, I. Lagzi, R. Horvath. Glycocalyx components detune the cellular uptake of gold nanoparticles in a size- and charge-dependent manner. ACS Applied Bio Materials, 2023.

[2] B. Peter, I. Lagzi, S. Teraji, H. Nakanishi, L. Cervenak, D. Zámbó, A. Deák, K. Molnár, M. Truszka, I. Szekacs, R. Horvath. Interaction of positively charged gold nanoparticles with cancer cells monitored by an in situ label-free optical biosensor and transmission electron microscopy. ACS Applied Materials & Interfaces, 2018.