The human micro-vascular endothelial cells in vitro interaction with atomic-nitrogen-doped diamond-like carbon thin films.

This paper reports the initial response of atomic nitrogen doped diamond like carbon (DLC) to endothelial cells in vitro. The introduction of nitrogen atoms/molecules to the diamond like carbon structures leads to an atomic structural change favorable to the attachment of human micro-vascular endothelial cells. Whilst the semi-conductivity induced by nitrogen in DLC is thought to play a part, the increase in the non-bonded N atoms and N(2) molecules in the atomic doped species (with the exclusion of the charged species) seems to contribute to the improved attachment of human microvascular endothelial cells. The increased endothelial attachment is associated with a lower work function and slightly higher water contact angle in the atomic doped films, where the heavy charged particles are excluded. The films used in the study were synthesized by the RF PECVD technique followed by post deposition doping with nitrogen, and afterwards the films were characterized by XPS, Raman spectroscopy, SIMS and Kelvin probe. The water contact angles were measured, and the counts of the adherent endothelial cells on the samples were carried out. This study is relevant and contributory to improving biocompatibility of surgical implants and prostheses.

Endothelial cell growth on silicon modified hydrogenated amorphous carbon thin films.

The biological response of human microvascular endothelial cells (HMEC-1) seeded on Si-DLC films and on control surfaces was evaluated in terms of initial cell enhancement, growth, and cytotoxicity. The microstructure of the films was characterised by Raman spectroscopy and X-ray photoelectron spectroscopy. The effect of changes in microstructure, surface energy, surface electronic state, and electronic conduction, on the biological response of the films to endothelial cells was investigated. Endothelial cell adhesion and growth was found to be affected by changes in the microstructure of the films induced by silicon doping and thermal annealing. We observed a significant statistical difference in endothelial cell count between the as-deposited DLC and Si-DLC films using the one sample t-test at a p-value of 0.05. We also found a statistically significant difference between the adhesion of HMEC films on DLC and Si-DLC films at various annealing temperatures using the one-way ANOVA F statistic test at p < 0.05 and the post-hoc Tukey test. One sample t-test at p < 0.05 of MTT-assay results showed the endothelial cells to be viable when seeded on DLC/Si-DLC films. We suspect that the increased adhesion of endothelial cells induced by increasing the amount of silicon in the Si-DLC films is associated with the development of a suitable surface energy due to silicon addition, which neither favored cell denaturing nor preferential water spreading before cellular attachment on the film surface. The presence of an external positively charged dipole on the Si-DLC films confirmed by our Kelvin probe measurements is also expected to enhance the adhesion of endothelial cells that are well known to carry a negative charge. The Si-DLC films investigated hold potential promise as coatings for haemocompatible artificial implants.

Enhancement of sp(3)-bonding in high-bias-voltage grown diamond-like carbon thin films studied by x-ray absorption and photoemission spectroscopy.

X-ray absorption near-edge structure (XANES) and valence-band photoemission spectroscopy (VB-PES) were used to elucidate the electronic and mechanical properties of diamond-like carbon (DLC) thin films deposited by the plasma-enhanced chemical vapour deposition method at various bias voltages (V(b)) using a C(2)H(2) vapour precursor in an Ar(+) atmosphere. The increase of V(b) is found to increase and decrease the contents of sp(3)- and sp(2)-bonded carbon atoms, respectively, i.e. the films become more diamond-like. The Young's modulus measurements show increases with the increase of the presence of sp(3)-bonded carbon atoms in the structure of the DLC films.

Human microvascular endothelial cellular interaction with atomic N-doped DLC compared with Si-doped DLC thin films.

This article reports results of endothelial cell interaction with atom beam source N-doped a-C:H (diamond-like carbon, DLC) as it compares with that of Si-doped DLC thin films. The RF plasma source exhibits up to 40% N-dissociation and N-atomic fluxes of approximately 0.85 x 10(18) atoms/s, which ensures better atomic nitrogen incorporation. Two different types of nitrogen species (with and without the use of sweep plates to remove charged ions) were employed for nitrogen doping. The number of attached endothelial cells is highest on Si-DLC, followed by the N-DLC (where the sweep plates were used to remove ions), the N-DLC (without the use of sweep plates), undoped DLC, and finally the uncoated sample. The contact angle values for these films suggest that water contact angle is higher in the atomic nitrogen neutral films and Si-DLC films compared to the ionized-nitrogen specie doped films and undoped DLC thin films, suggesting that the more hydrophobic films, semiconducting films, and film with relieved stress have better interaction with human microvascular endothelial cells. It seems evident that N-doping increases the Raman I(D)/I(G) ratios, whereas N-neutral doping decreases it slightly and Si-doping decreases it even further. In this study, lower Raman I(D)/I(G) ratios are associated with increased sp(3)/sp(2) ratio, an increased H concentration, photoluminescence intensity, and a higher endothelial cellular adhesion. These investigations could be relevant to biocompatibility assessment of nanostructured biomaterials and tissue engineering.

The MTT assays of bovine retinal pericytes and human microvascular endothelial cells on DLC and Si-DLC-coated TCPS.

MTT (Tetrazolium)-assay suggests that diamond-like carbon (DLC) and silicon-doped DLC (Si-DLC) films obtained under appropriate deposition parameters are not toxic to bovine retinal pericytes, and human microvascular endothelial cells (HMEC). The observed frequency distributions of the optical density (OD) values indicative of cell viability are near Gaussian-normal distribution. One-way ANOVA indicates that at 0.05 levels the population means are not significantly different for the coated and control samples. The observed OD values depend on the cell line (cell growth/metabolic rate), possibly cell cycle stage, the deposition parameters-bias voltage, ion energy, pressure, argon precleaning, and the dopant. For colored thin films like DLC with room temperature photoconductivity and photoelectric effects, it is important to account for the OD contribution from the coating itself. MTT assay, not surprisingly, seems not to be highly sensitive to interfacial cellular interaction resulting from the change in the film's nanostructure, because the tetrazolium metabolism is mainly intracellular and not interfacial. The thin films were synthesized by 13.56 MHz RF-PECVD using argon and acetylene as source gases, with tetramethylsilane (TMS) vapor introduced for silicon doping. This study could be relevant to biomedical application of the films in the eye, peri-vascular, vascular compartments, and for cell-tissue engineering.

Platelet adhesion on silicon modified hydrogenated amorphous carbon films.

We have investigated the effect of changes in microstructure, surface energy, surface charge condition and electronic conduction on the interaction of human platelets with silicon modified hydrogenated amorphous carbon films (a-C:H:Si or Si-DLC). Results based on Raman spectroscopy, Scanning electron microscopy, X-ray photo-electron spectroscopy, surface energy measurements, electrical resistivity, contact potential difference, and thermal annealing indicates a correlation between some of the measured values and the interaction of the films with human blood platelets. Statistical analysis of platelet aggregation on the films using the Student's t-test indicated differences between platelet aggregation on the modified films compared to the as-deposited film at a p-value of <0.05.