Roberto Gristina

Novel plasma processes for biomaterials: micro-scale patterning of biomedical polymers

Abstract Plasma processes of interest for tissue engineering, biosensors and related applications have been utilized for micro-scale patterning polymers with ‘physical masks’. The surface of polystyrene substrates has been patterned with different chemical micro-domains, each one able to induce different adhesion, spreading and growth behavior of cells. Cell adhesive tracks spaced with wider non-fouling PEO-like zones have been developed at the surface of the substrates, and utilized in cell growth experiments.

Cell Adhesion on Nanotextured Slippery Superhydrophobic Substrates

In this work, the response of Saos2 cells to polymeric surfaces with different roughness/density of nanometric dots produced by a tailored plasma-etching process has been studied. Topographical features have been evaluated by atomic force microscopy, while wetting behavior, in terms of water-surface adhesion energy, has been evaluated by measurements of drop sliding angle. Saos2 cytocompatibility has been investigated by scanning electron microscopy, fluorescent microscopy, and optical microscopy. The similarity in outer chemical composition has allowed isolation of the impact of the topographical features on cellular behavior. The results indicate that Saos2 cells respond differently to surfaces with different nanoscale topographical features, clearly showing a certain inhibition in cell adhesion when the nanoscale is particularly small. This effect appears to be attenuated in surfaces with relatively bigger nanofeatures, though these express a more pronounced slippery/dry wetting character.

Osteoblast-like cell behavior on plasma deposited micro/nanopatterned coatings.

The behavior of cells in terms of cell-substrate and cell-cell interaction is dramatically affected by topographical characteristics as shape, height, and distance, encountered in their physiological environment. The combination of chemistry and topography of a biomaterial surface influences in turns, important biological responses as inflammatory events at tissue-implant interface, angiogenesis, and differentiation of cells. By disentangling the effect of material chemistry from the topographical one, the possibility of controlling the cell behavior can be provided. In this paper, surfaces with different roughness and morphology were produced by radiofrequency (RF, 13.56 MHz) glow discharges, fed with hexafluoropropylene oxide (C(3)F(6)O), in a single process. Coatings with different micro/nanopatterns and the same uppermost chemical composition were produced by combining two plasma deposition processes, with C(3)F(6)O and tetrafluoroethylene (C(2)F(4)), respectively. The behavior of osteoblast-like cells toward these substrates clearly shows a strict dependence of cell adhesion and proliferation on surface roughness and morphology.

Phototoxicity and cytotoxicity of chlorophyll a/cyclodextrins complexes on Jurkat cells.

The aggregation status of chlorophyll a (Chl a) and the ability of four cyclodextrins, hydroxypropyl-beta-cyclodextrin (HP-beta-CD), hydroxypropyl-gamma-cyclodextrin (HP-gamma-CD), heptakis(2,6-di-O-methyl)-beta-cyclodextrin (DIMEB), and heptakis(2,3,6-tri-O-methyl)-beta-cyclodextrin (TRIMEB), to solubilize the pigment in the complete cellular medium RPMI 1640 was estimated by means of UV-Vis absorption and static resonance light scattering (RLS) measurements. The results indicate that the pigment interacts with cyclodextrins in the cellular medium differently to that observed in water. The cytotoxic and phototoxic activity of these complexes towards human leukemia T-lymphocytes (Jurkat cells) was tested by means of experiments aimed to discriminate between the intrinsic toxicity and the toxicity induced by light. The overall data indicate that the HP-beta-CD is the cyclodextrins having the best characteristics to form with Chl a a potential supramolecular system for the photodynamic therapy.

Biosilica from Living Diatoms: Investigations on Biocompatibility of Bare and Chemically Modified Thalassiosira weissflogii Silica Shells

In the past decade, mesoporous silica nanoparticles (MSNs) with a large surface area and pore volume have attracted considerable attention for their application in drug delivery and biomedicine. Here we propose biosilica from diatoms as an alternative source of mesoporous materials in the field of multifunctional supports for cell growth: the biosilica surfaces were chemically modified by traditional silanization methods resulting in diatom silica microparticles functionalized with 3-mercaptopropyl-trimethoxysilane (MPTMS) and 3-aminopropyl-triethoxysilane (APTES). Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy analyses revealed that the –SH or –NH2 were successfully grafted onto the biosilica surface. The relationship among the type of functional groups and the cell viability was established as well as the interaction of the cells with the nanoporosity of frustules. These results show that diatom microparticles are promising natural biomaterials suitable for cell growth, and that the surfaces, owing to the mercapto groups, exhibit good biocompatibility.

The study of specific and nonspecific hepatoma cells behavior by means of plasma‐treated substrates

Physical-chemical surface modifications represent a formidable tool to drive a suitable cell behavior on materials intended to be used in the biomedical field. Plasma processes are among the more powerful methods utilized to modify the surface of materials without altering their bulk intrinsic properties. In particular, by means of plasma treatment processes it is possible to graft chemical functional groups on polymer substrate. Functional groups grafted on the surface can improve per se cell adhesion and can also represent suitable anchor sites for biomolecule immobilization. The aim of this work was to determine the effect of plasma treatment and biomolecule immobilization on Polystyrene (PS) Petri dishes on the behavior of a human hepatocellular carcinoma cell line (HepG2). For this aim Petri dishes were grafted with N-containing groups in order to obtain grafted N-functionalities, to be used as anchor groups for the immobilization of galactosamine. In this way two different modified surfaces, NH(3) grafted polystyrene (PS-NH(3)) and polystyrene owing galactosamine moieties (PS-NH(3)-GalNH(2)), have been obtained. Differences in cell morphology, urea and plasma Fibronectin (pFN) production were clearly observed on HepG2 seeded on PS-NH(3) and PS-NH(3)-GalNH(2). These results highlight the role of specific and non specific cell response in the in vitro study of materials intended to be used for biomedical purposes.