title: In vitro biological responses to nanofibrillated cellulose by human dermal, lung and immune cells: surface chemistry aspect.
abstract: Nanocellulose, and particularly nanofibrillated cellulose (NFC), has been proposed for a diversity of applications in industry and in the biomedical field. Its unique physicochemical and structural features distinguish nanocellulose from traditional materials and enable its use as an advance nanomaterial. However, its nanoscale features may induce unknown biological responses. Limited studies with NFC are available and the biological impacts of its use have not been thoroughly explored. This study assesses the in vitro biological responses elicited by wood-derived NFC gels, when human dermal fibroblasts, lung MRC-5 cells and THP-1 macrophage cells are exposed to the nanomaterial. Furthermore, whether the presence of surface charged groups (i.e. carboxymethyl and hydroxypropyltrimethylammonium groups) on NFC can induce distinct biological responses is investigated.
  
  
  
  The introduction of surface charged groups resulted in individual nanofibrils, while fibril aggregates predominated in the unmodified NFC gel suspensions as observed by transmission electron microscopy. In the presence of proteins, the surface modified NFCs formed compact agglomerates while the agglomeration pattern of the unmodified NFC was similar in the presence of proteins and in physiological buffer. Unmodified and modified NFC gels did not induce cytotoxicity in human dermal fibroblasts, lung and macrophage cells. No significant ROS production by THP-1 macrophages was found and no cellular uptake was observed. However, an inflammatory response was detected when THP-1 macrophages were treated with unmodified NFC as assessed by an increase in TNF-α and IL1-β levels, an effect that was absent when surface charged groups were introduced into NFC.
  
  
  
  Taken together, the data presented here show the absence of cytotoxic effects associated with the exposure to unmodified, carboxymethylated and hydroxypropyltrimethylammonium-modified NFCs. Unmodified NFC presented a pro-inflammatory effect which can be further moderated by introducing surface modifications such as carboxymethyl and hydroxypropyltrimethylammonium groups into the nanofibrils. The present findings suggest that the inflammatory response to NFC might be driven by the material surface chemistry, and thus open up for the possibility of designing safe nanocellulose materials.
authors.affiliation: Nanotechnology and Functional Materials, Department of Engineering Sciences, Uppsala University, Box 534, 75121, Uppsala, Sweden. viviana.lopes@angstrom.uu.se.
authors.forename: Viviana R
authors.initials: VR
authors.lastname: Lopes
authors.affiliation: Present affiliation: Ocular Biology and Therapeutics, UCL Institute of Ophthalmology, 11-43 Bath Street, EC1V 9EL, London, UK.
authors.forename: Carla
authors.initials: C
authors.lastname: Sanchez-Martinez
authors.affiliation: Nanotechnology and Functional Materials, Department of Engineering Sciences, Uppsala University, Box 534, 75121, Uppsala, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Strømme
authors.affiliation: Nanotechnology and Functional Materials, Department of Engineering Sciences, Uppsala University, Box 534, 75121, Uppsala, Sweden. Natalia.Ferraz@angstrom.uu.se.
authors.forename: Natalia
authors.initials: N
authors.lastname: Ferraz
mesh_terms: D002460:Cell Line; D002482:Cellulose / Q000737:chemistry / Q000633:toxicity*; D006801:Humans; D066298:In Vitro Techniques; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008264:Macrophages / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D057139:Nanofibers / Q000737:chemistry / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D002482:Cellulose; D007770:L-Lactate Dehydrogenase
keywords: Cytotoxicity; Inflammation; Nanocellulose; Surface chemistry