The last decade has seen an impressive increase of the research dedicated to the biomedical applications of low temperature Non Thermal Plasmas (ltNTP), especially with plasma sources working at atmospheric pressure. Medical applications of ltNTP now concern a very wide range of domains including cancer treatment. The antitumor effect of ltNTP has been clearly shown in vivo on murine modelswith various cancer types (bladder, colon, glioblastoma, melanoma, ovary, pancreas). Although the involved mechanisms are far from being fully understood, the therapeutic effect is now totally admitted and the first clinical study (head and neck) has been reported [1]. In case of plasma jet experiments, the observed effect are most of the time attributed to the very rich chemistry generated by the interaction of the rare gas plasma plume with the surrounding environment constituted either from the ambient air, or this latter in complex interaction with liquids at the interface with the targeted organ. Our recent experiments performed on tissue oxygenation[2] or breast cancer treatments on immunocompetent mice [3] lead to the conclusion that probably the involved chemistry couldn’t, alone, completely allow describing the observed phenomena. This, especially under very soft treatment conditions, is suggesting possible triggering of some immune system chain processes and also possible modifications in the microenvironment of tissue and tumors. In this context, there is still an unknown role of the electric field associated with the ionization front or generated in the environment of the plasma plume tip. Taking into consideration the recent vessel normalization based-cancer treatment, the ltNTP effect should be further investigated in view of blood vessels structure and function (blood flow) as well as tumor hypoxia compensation to confirm a possible ltNTP-based adjuvant approach for cancer treatments. These results suggest new ways, especially combined therapy, to consider the plasma and its therapeutic delivery in ltNTP-based tumor therapy. In this talk, after a presentation of the context and the plasma devices, we will go through the specific case of cancer treatment with what have been already demonstrated in vitro and in vivo, what can be directly linked with the produced discharges, including recent results on electric field measurements in plasma biological application conditions.

This work is supported by the APR Region Centre PLASMEDNORM.

References:

[1] H.R. Metelmann et al Clin. Plas Med. Doi.org/10.1016/j.cpme.2015.02.001

[2] G. Collet et al PSST 23 (2014) 012005

[3] G. Collet et al ICPM5, May 18-23, 2014, Nara (Japan)

Thin film metallic glasses (TFMGs) exhibit unique properties such as high strength, smooth surface as well as good wear- and corrosion-resistances due to their amorphous atomic structure. The biocompatibility and antibacterial property of TFMGs can also be obtained, which show great potential for biomedical applications. In addition, the low surface free energy of TFMGs in certain compositions can be achieved and leads to the relatively high hydrophobicity and the low friction coefficient.

In this presentation, various applications of TFMG are discussed, including the property enhancements of dermatome blade and syringe needle, thrombosis reduction for intravenous catheter, and the suppression of cancer cell attachments. A Zr-based TFMG is coated on substrates by using magnetron sputtering. The TFMG-coated dermatome blade show a great enhancement of durability and sharpness, compared with those of the bare one. For the syringe needle, significant reductions in insertion and retraction forces for TFMG-coated needle are achieved due to the non-sticky property and relatively low coefficient of friction. For thrombosis reduction, less platelet aggregations are observed on the TFMG than that on the bare glass in platelets adhesion test, suggesting TFMG-coated catheters is potentially useful to be placed into vessels for long periods of time with reduced numbers of the aggregation of blood platelets. For cancer cell attachment suppressions, TFMG exhibits the least cancer cell attachment among other control groups. Thus, anti-proliferation and anti-metastasis of medical tools can be achieved with TFMG coating.

In this study, we will present the recent experimental results on plasma surface functionalization of nano-structured materials for bio-medical applications.

First, with the graphite-encapsulated magnetic nanoparticles(MNPs), we studied the surface functionalization by using the Ar plasma pre-treatment followed by NH₃ plasma post-treatment, to introduce the amino groups onto the surface of the nanoparticles.¹⁾ The amino group population of each nanoparticle with a typical diameter of 20 nm was evaluated by using the conventional chemical technique using SPDP and DTT solutions and we obtained about 8 x 10⁴ amino groups per nanoparticle.²⁾ Immobilization of the antibody of influenza virus onto the surface of amino-modulated magnetic nanoparticles was then performed for aiming at studying the feasibility of collection and condensation of virus. After magnetic separation, we succeeded in a significant concentration of the influenza virus number compared with that of the initial sample.³⁾ Using the same method, we also demonstrated a higher concentration of Salmonella about 70 times higher than that of initial sample by the magnetic separation.⁴⁾ The present results suggest the feasibility of the proposed plasma surface functionalized MNPs for rapid concentration of influenza virus or various bacteria.

As the second topic, the selective ultrafine surface modification of functional groups onto the polymeric substrate or vertically aligned CNT dot-array with a dot size of several μm was investigated using the atmospheric pressure plasma jet with a nano/micro-sized capillary. The micro-sized surface modification of amino or carboxyl groups introduced onto the CNT dot-array were confirmed by the fluorescence labelling technique.⁵⁾ With fluorescence-labeled avidin molecules, we also confirmed efficient capturing of avidin molecules by the biotin-immobilized CNT dot array through strong biotin-avidin binding process. The present result supports the feasibility of future biochip sensor to detect specific protein, virus or bacteria. In addition to these results, the other experimental results will be presented and discussed at the conference.

References

1) T. E. Saraswati, A. Ogino, M. Nagatsu, Carbon, 50 (2012) pp.1253-1261.

2) T. E. Saraswati, S. Tsumura, and M. Nagatsu, Jpn. J. Appl. Phys. 53 (2014) 010205(5 pages).

3) A. Sakudo, H. Chou, K. Ikuta, and M. Nagatsu, Bioorg. Med. Chem. Lett. 25 (2015) pp.1876–1879.

4) A. Sakudo, H. Chou, and M. Nagatsu, Bioorg. Med. Chem. Lett., 25 (2015) pp. 1012-1016

5) T. Abuzairi, M. Okada, Y. Mochizuki, N. R. Poespawati, R. Wigajatri and M. Nagatsu, Carbon, 89 (2015) pp. 208-216.

The immobilization of biologically active species is crucial for the fabrication of smart bioactive surfaces. For this purpose, plasma polymerization is frequently used to modify the surface nature without affecting the bulk properties of the material. Thus, it is possible to create materials with surface functional groups that can promote the anchoring of all kinds of biomolecules. Different methodologies in protein immobilization have been developed in recent years, although some drawbacks are still not solved, such as the difficulties that some procedures involve and/or the denaturalization of the protein due to the immobilization process. However, along with the chemical signals, the mechanical forces are critical for many tissues, since they are constantly suffering tension, shear, loading, etc. Essentially, the cell signaling exerted by forces is transduced through receptors that are in intimate contact with the matrix. Therefore, the main consequence of this receptor-matrix interaction is that cells and matrix are mechanically coupled, so that matrix deformation is considered the main cause of the mechanical signaling. By mimicking these mechanical forces in the surface of a material, it would be possible to obtain more physiological environments and thus a more physiological cell response. Again, the use of plasma polymerization techniques can help to design surfaces that can be tailored in terms of mechanical properties and chemical compositions and thus have a high potential for cells signaling.

This paper reports the work that we have developed in the last 10 years in the design, synthesis and characterization of thin films that can be a platform for studying the interaction between cells and separate influences of physical and chemical cues of a matrix on the adhesion, growth and final phenotype of cells.

Polystyrene is a widely used cell-culture plate material. Currently cell culture plates on the market include those whose inner surfaces are covered with amino and/or carbonyl groups for a better control of cell adhesion to the plate surfaces. Such functional groups on a cell culture plate surface may immobilize glycoproteins or other biopolymers that function as extracellular matrices (ECM) and thus affect the environments where the cells are cultured. The goal of this research is to understand how such functional groups, especially amino groups, are formed on a polystyrene surface, depending on the deposition methods. Of particular interest are plasma-based methods of surface functionalization. In this study, we have observed experimentally how exposure of N₂/H₂ or N₂/CH₃OH plasmas to polystyrene surfaces form amino-group-like structures and also examined using molecular dynamics (MD) simulation how a polystyrene surface interacts with incident energetic ions such as NH₃⁺ as well as abundant low-energy radicals such as NH₂ under conditions similar to our experiments. In the experiments, we used parallel-plate discharges with an inverter power supply whose peak-to-peak voltage was about 3kV and frequency was 20kHz at a relatively high gas pressure of 250 - 2,500 Pa. In MD simulation, we used a simulation code with interatomic potential functions that had been developed in-house based on quantum mechanical calculations of atomic interactions involved in this system. Results of MD simulations under the conditions similar to plasma enhanced chemical vapor deposition (PE-CVD) by ammonia plasmas or cyclopropylamine (CPA) [1] suggest that, with energetic ion bombardment, amino groups tend to be broken to form new covalent bonds by ion bombardment. Preliminary results of cell culture experiments with plasma-treated polystyrene cell plates will be also reported.

[1] A. Manakhov, L. Zajickova, et al. Plasma Process. Polym.11, (2014) 532.

Utilizing bioresorbable polymers to fabricate constructs with three-dimensional (3D), porous architectures is desirable as these constructs mimic the extracellular matrix– a critical characteristic for many biomedical applications including tissue engineering, controlled-release drug delivery, and wound healing. Although the bioresorbability and architecture of these materials are suitable for such applications, the surface properties (i.e., chemical functionality and wettability) must often be customized depending on the desired function. Plasma processing is an attractive tool for surface modification of these delicate polymeric materials as it provides a low-temperature, sterile environment with a variety of precursor choices. The presented work will highlight the plasma modification of a variety of 3D, porous polymeric constructs. Specifically, we fabricated scaffolds via electrospinning and porogen leaching techniques using both poly(ε-caprolactone) (PCL) and polylactic acid (PLA) to develop a repertoire of native polymer constructs with differing bulk properties. We evaluated the efficacy of plasma-modifying 3D constructs using contact angle goniometry, X-ray photoelectron spectroscopy, and scanning electron microscopy to assess changes in wettability, chemical functionality, and scaffold architecture. The interactions of plasma-modified scaffolds with different biological species, including human dermal fibroblasts and Escherichia coli were explored, specifically to assess scaffold bioreactivity. Notably, we demonstrate that scaffold properties, and thus bioreactivity, can be customized depending on the choice of plasma precursor. We show that plasma treatment using fluorocarbon and hydrocarbon precursors (i.e., octofluoropropane, hexafluoropropylene oxide, and 1,7-octadiene) results in hydrophobic and bio-non reactive scaffolds. Additionally, precursors with nitrogen and oxygen functionality (i.e., allylamine, allyl alcohol, water, and ammonia) can be used to fabricate scaffolds that are hydrophilic and bio-reactive. Altogether, this work illustrates the comprehensive tunability of biologically-relevant polymeric constructs in terms of their bulk properties, surface properties, and cell-surface interactions.