Nanomaterials have unusual properties not found in the bulk materials, which can be exploited in numerous applications such as biosensing, electronics, scaffolds for tissue engineering, diagnostics and drug delivery. However, research in the past few years has turned up a range of potential health hazards, which has given birth to the new discipline of nanotoxicology. Bacterial cellulose (BC) is a promising material for biomedical applications, namely due its biocompatibility. Although BC has been shown not to be cytotoxic or genotoxic, the properties of isolated BC nanofibres (NFs) on cells and tissues has never been analysed. Considering the toxicity associated to other fibre-shaped nanoparticles, it seems crucial to evaluate the toxicity associated to the BC-NFs. In this work, nanofibres were produced from bacterial cellulose by a combination of acid and ultrasonic treatment. The genotoxicity of nanofibres from bacterial cellulose was analysed in vitro, using techniques previously demonstrated to detect the genotoxicity of fibrous nanoparticles. The results from single cell gel electrophoresis (also known as comet assay) and the Salmonella reversion assays showed that NFs are not genotoxicity under the conditions tested. A proliferation assay using fibroblasts and CHO cells reveals a slight reduction in the proliferation rate, although no modification in the cell morphology is observed.