Safe subdural administration and retention of a neurotrophin-3-delivering hydrogel in a rat model of spinal cord injury.

Neurotrophic growth factor (GF) loaded hydrogels have shown promise as a treatment approach for spinal cord injury (SCI). However, SCI presents complex challenges for the direct administration of treatment due to the spinal cord's intricate anatomy and highly sensitive environment. Many current hydrogel administration approaches overlook this complexity, limiting their translational potential.

Encapsulation of the growth factor neurotrophin-3 in heparinised poloxamer hydrogel stabilises bioactivity and provides sustained release.

Poloxamer-based hydrogels show promise to stabilise and sustain the delivery of growth factors in tissue engineering applications, such as following spinal cord injury. Typically, growth factors such as neurotrophin-3 (NT-3) degrade rapidly in solution. Similarly, poloxamer hydrogels also degrade readily and are, therefore, only capable of sustaining the release of a payload over a small number of days.

Translating ultrasound-mediated drug delivery technologies for CNS applications.

Ultrasound enhances drug delivery into the central nervous system (CNS) by opening barriers between the blood and CNS and by triggering release of drugs from carriers. A key challenge in translating setups from in vitro to in vivo settings is achieving equivalent acoustic energy delivery. Multiple devices have now been demonstrated to focus ultrasound to the brain, with concepts emerging to also target the spinal cord.

Generation of direct current electrical fields as regenerative therapy for spinal cord injury: A review.

Electrical stimulation (ES) shows promise as a therapy to promote recovery and regeneration after spinal cord injury. ES therapy establishes beneficial electric fields (EFs) and has been investigated in numerous studies, which date back nearly a century. In this review, we discuss the various engineering approaches available to generate regenerative EFs through direct current electrical stimulation and very low frequency electrical stimulation.

Investigating the influence of ultrasound parameters on ibuprofen drug release from hydrogels.

Hydrogels are promising ultrasound-responsive drug delivery systems. In this study, we investigated how different ultrasound parameters affected drug release and structural integrity of self-healing hydrogels composed of alginate or poloxamers. The effects of amplitude and duty cycle at low frequency (24 kHz) ultrasound stimulation were first investigated using alginate hydrogels at 2% w/v and 2.

Patternable Gelatin Methacrylate/PEDOT/Polystyrene Sulfonate Microelectrode Coatings for Neuronal Recording.

This manuscript addresses the need for new soft biomaterials that can be fabricated on the surface of microelectrodes to reduce the mechanical mismatch between biological tissues and electrodes and improve the performance at the neural interface. By electrochemical polymerization of poly(3,4-dioxythiophene) (PEDOT)/polystyrene sulfonate (PSS) through a gelatin methacrylate (GelMA) hydrogel, we demonstrate the synthesis of a conducting polymer hydrogel (CPH) to meet the performance criteria of bioelectrodes. The hybrid material can be photolithographically patterned and covalently attached to gold microelectrodes, forming an interpenetrating network, as confirmed by infrared spectroscopy.

Development of agarose-gelatin bioinks for extrusion-based bioprinting and cell encapsulation.

Three-dimensional bioprinting continues to advance as an attractive biofabrication technique to employ cell-laden hydrogel scaffolds in the creation of precise, user-defined constructs that can recapitulate the native tissue environment. Development and characterisation of new bioinks to expand the existing library helps to open avenues that can support a diversity of tissue engineering purposes and fulfil requirements in terms of both printability and supporting cell attachment. In this paper, we report the development and characterisation of agarose-gelatin (AG-Gel) hydrogel blends as a bioink for extrusion-based bioprinting.

Conducting polymer hydrogels with electrically-tuneable mechanical properties as dynamic cell culture substrates.

Hydrogels are a popular substrate for cell culture due to their mechanical properties closely resembling natural tissue. Stimuli-responsive hydrogels are a good platform for studying cell response to dynamic stimuli. Poly(N-isopropylacrylamide) (pNIPAM) is a thermo-responsive polymer that undergoes a volume-phase transition when heated to 32 °C.

A Subdural Bioelectronic Implant to Record Electrical Activity from the Spinal Cord in Freely Moving Rats.

Bioelectronic devices have found use at the interface with neural tissue to investigate and treat nervous system disorders. Here, the development and characterization of a very thin flexible bioelectronic implant inserted along the thoracic spinal cord in rats directly in contact with and conformable to the dorsal surface of the spinal cord are presented. There is no negative impact on hind-limb functionality nor any change in the volume or shape of the spinal cord.

Optimised techniques for high-throughput screening of differentiated SH-SY5Y cells and application for neurite outgrowth assays.

Neuronal models are a crucial tool in neuroscientific research, helping to elucidate the molecular and cellular processes involved in disorders of the nervous system. Adapting these models to a high-throughput format enables simultaneous screening of multiple agents within a single assay. SH-SY5Y cells have been widely used as a neuronal model, yet commonly in an undifferentiated state that is not representative of mature neurons.

An interpenetrating and patternable conducting polymer hydrogel for electrically stimulated release of glutamate.

Recent advances in drug delivery have made it possible to release bioactive agents from neural implants specifically to local tissues. Conducting polymer coatings have been explored as a delivery platform in bioelectronics, however, their utility is restricted by their limited loading capacity and stability. This study presents the fabrication of a stable conducting polymer hydrogel (CPH), comprising the hydrogel gelatin methacrylate (GelMA), and conducting polymer polypyrrole (PPy) for the electrically controlled delivery of glutamate (Glu).

Stretchable microchannel-on-a-chip: A simple model for evaluating the effects of uniaxial strain on neuronal injury.

Background: Axonal injury is a major component of traumatic spinal cord injury (SCI), associated with rapid deformation of spinal tissue and axonal projections. In vitro models enable us to examine these effects and screen potential therapies in a controlled, reproducible manner.

New Method: A customized, stretchable microchannel system was developed using polydimethylsiloxane microchannels.

A Macroscopic Diffusion-Based Gradient Generator to Establish Concentration Gradients of Soluble Molecules Within Hydrogel Scaffolds for Cell Culture.

Concentration gradients of soluble molecules are ubiquitous within the living body and known to govern a number of key biological processes. This has motivated the development of numerous gradient-generators allowing researchers to study cellular response in a precise, controlled environment. Despite this, there remains a current paucity of simplistic, convenient devices capable of generating biologically relevant concentration gradients for cell culture assays.

Evaluation of parylene derivatives for use as biomaterials for human astrocyte cell patterning.

Cell patterning is becoming increasingly popular in neuroscience because it allows for the control in the location and connectivity of cells. A recently developed cell patterning technology uses patterns of an organic polymer, parylene-C, on a background of SiO2. When cells are cultured on the parylene-C/SiO2 substrate they conform to the underlying parylene-C geometry.

Selective PEGylation of Parylene-C/SiO Substrates for Improved Astrocyte Cell Patterning.

Controlling the spatial distribution of glia and neurons in in vitro culture offers the opportunity to study how cellular interactions contribute to large scale network behaviour. A recently developed approach to cell-patterning uses differential adsorption of animal-serum protein on parylene-C and SiO surfaces to enable patterning of neurons and glia. Serum, however, is typically poorly defined and generates reproducibility challenges.

Patterning of functional human astrocytes onto parylene-C/SiO substrates for the study of Ca dynamics in astrocytic networks.

Objective: Recent literature suggests that astrocytes form organized functional networks and communicate through transient changes in cytosolic Ca. Traditional techniques to investigate network activity, such as pharmacological blocking or genetic knockout, are difficult to restrict to individual cells. The objective of this work is to develop cell-patterning techniques to physically manipulate astrocytic interactions to enable the study of Ca in astrocytic networks.

Nanosecond UV lasers stimulate transient Ca elevations in human hNT astrocytes.

Objective: Astrocytes respond to various stimuli resulting in intracellular Ca signals that can propagate through organized functional networks. Recent literature calls for the development of techniques that can stimulate astrocytes in a fast and highly localized manner to emulate more closely the characteristics of astrocytic Ca signals in vivo.

Approach: In this article we demonstrate, for the first time, how nanosecond UV lasers are capable of reproducibly stimulating Ca transients in human hNT astrocytes.

Investigation of the Ca2+ response of human hNT astrocytes to laser removal of cellular processes.

We demonstrate, for the first time, UV laser ablative microsurgery as a method for pruning astrocytic processes from live hNT astrocytic networks in vitro. Calcium fluorescence imaging was used to evaluate the cellular response to process ablation. The results showed that ablation of astrocyte processes induced an immediate increase in intracellular calcium level which propagated through the cells cytoplasm as a wave originating from the ablation site.

Investigating parylene-HT as a substrate for human cell patterning.

We demonstrate, for the first time, how parylene-HT on SiO2 substrates can be used as a human cell patterning platform. We demonstrate this platform with hNT astrocytes, derived from the human NTera2.D1 cell line.

Human astrocytic grid networks patterned in parylene-C inlayed SiO2 trenches.

Recent literature suggests that glia, and in particular astrocytes, should be studied as organised networks which communicate through gap junctions. Astrocytes, however, adhere to most surfaces and are highly mobile cells. In order to study, such organised networks effectively in vitro it is necessary to influence them to pattern to certain substrates whilst being repelled from others and to immobilise the astrocytes sufficiently such that they do not continue to migrate further whilst under study.

Low cost, patterning of human hNT brain cells on parylene-C with UV & IR laser machining.

This paper describes the use of 800nm femtosecond infrared (IR) and 248nm nanosecond ultraviolet (UV) laser radiation in performing ablative micromachining of parylene-C on SiO2 substrates for the patterning of human hNT astrocytes. Results are presented that support the validity of using IR laser ablative micromachining for patterning human hNT astrocytes cells while UV laser radiation produces photo-oxidation of the parylene-C and destroys cell patterning. The findings demonstrate how IR laser ablative micromachining of parylene-C on SiO2 substrates can offer a low cost, accessible alternative for rapid prototyping, high yield cell patterning.

Infra-red laser ablative micromachining of parylene-C on SiO2 substrates for rapid prototyping, high yield, human neuronal cell patterning.

Cell patterning commonly employs photolithographic methods for the micro fabrication of structures on silicon chips. These require expensive photo-mask development and complex photolithographic processing. Laser based patterning of cells has been studied in vitro and laser ablation of polymers is an active area of research promising high aspect ratios.

[The role of large wildlife in the maintenance of natural foci of tick-borne meningoencephalitis in northern Croatia].

Between 1986-1988 sera from 245 game animals--37 roe deers, 102 red deers, 81 wild boars and 25 hares--shot in 8 different localities of Nort Croatia, a well-known nosoareal of tick-borne encephalitis, were tested for TBE virus heminhibiting antibodies. The following percentages of positive animals were found: 24, 39, 39 and 0% respectively. Most of the animals examined, 180 of them, were shot in lowland woods of the commune of Nasice and its surroundings.