In vitro cytotoxicity of amorphous carbon films

Amorphous carbon (a-C), carbon nitride (a-CN) and titanium films were deposited on stainless steel substrates (SS) using a dc magnetron sputtering system attached to a high vacuum chamber. Films were deposited using a base pressure of 1.3x10(-4) Pa. For the carbon films a pure graphite target was eroded in an Argon plasma. For the case of the a-CN films, the Ar flux was substituted by 100% N2 gas. Titanium films were deposited in a different chamber, using a pure Ti target and an argon plasma. In vitro studies were carried out on the coated samples using human osteoblasts cells. Cytotoxicity of carbon films was assessed by cellular adhesion and proliferation, as determined by direct cellular counting using a spectroscopic technique and a well-defined standard curve. Osteoblasts cells were also grown on uncoated steel and prepared Petri dishes for comparison. The percentage of osteoblasts adhesion measured at 24 hrs attained maximum values for the a-C films. Similarly, cellular proliferation evaluated at three, five and seven days showed an outstanding increase of osteoblasts cells for the a-C and Ti coatings in contrast to the uncoated steel. The cell functionality was evaluated by the MTT test after incubation periods of 3, 5 and 7 days. The absorbance values obtained for a-C, a-CN and Ti surfaces resulted significantly higher with respect to the positive control, indicating that the surface did not induce any toxic effect. Preliminary bio-mineralization was evaluated by measuring the elemental composition of the mineral grown on the substrates after periods up to 14 days.