Electrically conductive bacterial cellulose for tissue-engineered neural interfaces

Bacterial cellulose (BC) with its high crystallinity, tensile strength, degree of polymerisation, and water holding capacity (98%) becomes increasingly attractive as 3D nanofibrillar material for biomedical applications. Such multi-scale fibrillary BC networks can be potentially functionalised with electrically conductive moieties to facilitate the conductive properties required for various smart biomedical devices, in particular, in the construction of bioelectronic neural interfaces. In this thesis, BC fibres are chemically modified with poly(4-vinylaniline) (PVAN) interlayer for further enhancement of electrical conductivity and cell viability of subsequent polyaniline (PANI) coatings as a bilayer grafted BC nanocomposite. This functional poly(4-vinylaniline)/polyaniline (PVAN/PANI) bilayer can be efficiently anchored onto BC fibrils through successive surface-initiated atom transfer radical polymerisation and in situ chemical oxidative polymerisation. PVAN is found to have promoted the formation of a uniform PANI layer with 1D nanofiber- and nanorod-like supramolecular structures, with an overall augmentation of PANI yield, hence further improved electrical performance. Compositional and microstructural analysis reveals such a PVAN/PANI bilayer with a thickness up to ~2 µm on BC formed through a significant growth of PANI with rough surface morphology due to the insertion of PVAN, which has improved the functional properties of the BC nanocomposites. Successful impregnation of both layers onto BC fibrils was corroborated with systematic microstructural and chemical analysis. The solid-state electrical conductivity of such synthesised BC nanocomposites with PVAN interlayer reaches as high as (4.5±2.8)×10-2 S.cm-1 subject to the amounts of PVAN chemically embraced. Electrochemical examination evinces the switching in the electrochemical behaviour of BC/PVAN/PANI nanocomposites at -0.70/0.74 V (at 100 mV.s-1 scan rate) due to the existence of PANI, where the maximal electrical performance can be achieved at charge transfer resistance of as low as 21 Ω and capacitance of as high as 39 μF. Both electrochemical and mechanical properties can be tailored onto an incomplete BC dehydration, where a mathematical model is herein developed to predict BC water loss accordingly. BC/PVAN/PANI nanocomposites are thermally stable up to 200 ºC. Furthermore, further improvement of the electrical conductivity has been achieved through grafting Carbon Nano Tubes (CNTs) into the BC/PVAN/PANI nanocomposites, where the interactions between PANI and CNTs present new electrochemical characteristics with enhanced capacity. PANI/CNTs coatings with a nanorod-like morphology can promote the efficient ions diffusion and charge transfer, resulting in the increased electrical conductivity up to (1.0±0.3)×10-1 S.cm-1. An escalating amplification of the double charge capacity to ~54 mF of the CNTs grafted BC nanocomposites was also detected through electrochemical analysis. In addition, the thermal stability of CNTs grafted BC/PVAN/PANI nanocomposites are improved, and they become stable up to 234 ºC. Cytocompatibility tests conducted using two neuronal cell linages show non-cytotoxic effects for PC-12 Adh cells and SVZ neural stem cells, confirming cell viability that can be over 80 % and neuronal differentiation capability of the electrically functionalised BC-based nanocomposite membranes, which can induce neurites outgrowth up to 115±24 μm long. These voltage-sensible nanocomposites can hence interact with neural cells, thereby significantly stimulate specialised response. These findings pave the path to the new tissue engineered neural interfaces which embraces electronic functions into the tissue regeneration, to enable full functional neural tissue recovery

Poly(4-vinylaniline)/polyaniline bilayer functionalized bacterial cellulose membranes as bioelectronics interfaces

Bacterial cellulose (BC) fibers are chemically functionalized with poly(4-vinylaniline) (PVAN) interlayer for further enhancement of electrical conductivity and cell viability of polyaniline (PANI) coated BC nanocomposites. PVAN is found to have promoted the formation of a uniform PANI layer with nanofiber- and nanorod-like supramolecular structures, as an overall augmentation of PANI yield. Compositional and microstructural analysis indicates a PVAN/PANI bilayer of approximately 2 μm formed on BC. The solid-state electrical conductivity of such synthesized BC nanocomposites can be as high as (4.5 ± 1.7) × 10−2 S cm−1 subject to the amounts of PVAN chemically embraced. BC/PVAN/PANI nanocomposites are confirmed to be thermally stable up to 225 °C, and no signs of cytotoxicity for SVZ neural stem cells are detected, with cell viability up to 90% on BC/PVAN/PANI membranes. We envisage these new electrically conductive BC/PVAN/PANI nanocomposites can potentially enable various biomedical applications, such as for the fabrication of bioelectronic interfaces and biosensors

Carbon nanotube-reinforced poly(4-vinylaniline)/polyaniline bilayer-grafted bacterial cellulose for bioelectronic applications

Microbial cellulose paper treated with polyaniline and carbon nanotubes (PANI/CNTs) can be attractive as potential flexible capacitors in terms of further improvements to the conductivity and thermal resistance. The interactions between PANI and CNTs exhibit new electrochemical features with increased electrical conductivity and enhanced capacity. In this study, PANI/CNTs was incorporated into a flexible poly(4-vinylaniline)-grafted bacterial cellulose (BC/PVAN) nanocomposite substrate for further functionalization and processability. PANI/CNTs coatings with a nanorod-like structure can promote an efficient ion diffusion and charge transfer, with a significant enhancement of the electrical conductivity after CNTs reinforcement of 1 order of magnitude up to (1.0 ± 0.3) × 10−1 S·cm−1 . An escalating improvement of the double charge capacity (∼54 mF) of the grafted BC nanocomposites was also detected through electrochemical analysis. The multilayered electrical coatings also reinforce the thermal resistance, preventing anticipated thermal degradation of the BC substrate. The cell viability and differentiation assays using neural stem cells (SVZ cells) testified to the cytocompatibility of the grafted BC nanocomposites, while inducing neuronal differentiation over 7 days of culture with a neurite that was 77 ± 24.7 μm long. This is promising for meeting the requirements in the construction of high-performance bioelectronic devices that can actively interface biologically, providing a friendly environment for cells while tuning the device performance

Poly(4-vinylaniline)/polyaniline bilayer functionalized bacterial cellulose for flexible electrochemical biosensors

Bacterial cellulose (BC) nanofibril network is modified with an electrically conductive polyvinylaniline/polyaniline (PVAN/PANI) bilayer for construction of potential electrochemical biosensors. This is accomplished through surface-initiated atom transfer radical polymerization of 4-vinylaniline, followed by in situ chemical oxidative polymerization of aniline. A uniform coverage of BC nanofiber with 1D supramolecular PANI nanostructures is confirmed by FTIR, XRD and CHN elemental analysis. Cyclic voltammograms evince the switching in the electrochemical behavior of BC/PVAN/PANI nanocomposites from the redox peaks at 0.74 V, in the positive scan and at -0.70 V, in the reverse scan, (at 100 mV.s-1 scan rate). From these redox peaks, PANI is the emeraldine form with the maximal electrical performance recorded, showing charge-transfer resistance as low as 21 Ω and capacitance as high as 39 μF. The voltage-sensible nanocomposites can interact with neural stem cells (NSCs) isolated from subventricular zone (SVZ) of the brain, through stimulation and characterization of differentiated SVZ cells into specialized and mature neurons with long neurites measuring up to 115±24 μm length after 7 days of culture without visible signs of cytotoxic effects. The findings pave the path to the new effective nanobiosensor technologies for nerve regenerative medicine, which demands both electroactivity and biocompatibility