In Vitro Biocompatibility Test of Multiwall Carbon Nanotubes with Human Osteoblast Cells: Potential Application for Bone Implant Interface Reinforcement

Abstract

Application of multiwall carbon nanotubes (MWCNT) as a filler component in composite materials can lead to remarkable increase in mechanical strength. It is a challenging application to form a living bone tissue biocomposite that is reinforced with MWCNTs at a dental implant—bone interface. The successful biointegration of MWCNT and the implant material depends on the processes of osseointegration, namely surface interactions at the molecular and cellular level. In this work the compatibility of MWCNT with main osseointegration processes has been overviewed with special attention to the toxicity of MWCNT for interacting human cells, and In Vitro experiments were performed with primary human osteoblast cells. The cells were isolated from oral bone fragments and grown in cell culture conditions. Plate wells were covered with MWCNT layers of three different densities. Osteoblast cell suspensions were placed onto the MWCNT layers and into empty plate wells. 24 and 72 hours after seeding the attachment and proliferation of cells was evaluated using Thiazolyl Blue Tetrazolium Bromide (MTT) colorimetric assay. The extent of cell death was characterized by Lactate Dehydrogenase (LDH) assay. The osteoblast cell viability tests show that cells were attached to all investigated surfaces, but with lower rate to higher density MWCNTs. A low level of cell death was observed in each sample type. Phase contrast and fluorescent microscopic observations show that although MWCNTs are not toxic for human primary osteoblast cells, an intense interaction of the cells with MWCNTs reduces their proliferation and markedly affects their morphology.