Review of in vitro mechanical testing for intervertebral disc injectable biomaterials.

Many early stage interventions for intervertebral disc degeneration are under development involving injection of a biomaterial into the affected tissue. Due to the complex mechanical behaviour of the intervertebral disc, there are challenges in comprehensively evaluating the performance of these injectable biomaterials in vitro. The aim of this review was to examine the different methods that have been developed to mechanically test injectable intervertebral disc biomaterials in an in vitro disc model. Testing methods were examined with emphasis on overall protocol, artificial degeneration method, mechanical testing regimes and injection delivery. Specifically, the effects of these factors on the evaluation of different aspects of device performance was assessed. Broad testing protocols varied between studies and enabled evaluation of different aspects of an injectable treatment. Studies employed artificial degeneration methodologies which were either on a macro scale through mechanical means or on a microscale with biochemical means. Mechanical loading regimes differed greatly across studies, with load being either held constant, ramped to failure, or applied cyclically, with large variability on all loading parameters. Evaluation of the risk of herniation was possible by utilising ramped loading, whereas cyclic loading enabled the examination of the restoration of mechanical behaviour for initial screening of biomaterials and surgical technique optimisation studies. However, there are large variations in the duration or tests, and further work is needed to define an appropriate number of cycles to standardise this type of testing. Biomaterial delivery was controlled by set volume or haptic feedback, and future investigations should generate evidence applying physiological loading during injection and normalisation of injection parameters based on disc size. Based on the reviewed articles and considering clinical risks, a series of recommendations have been made for future intervertebral disc mechanical testing.

Spatial localization of electrotactile stimuli on the fingertip in humans.

This study was designed to determine the extent to which sensations elicited by discrete electrotactile stimulation can be spatially localized, with a qualitative comparison to mechanical stimulation, in a 2 x 2 electrode array on the fingertip. Electrotactile stimulation was delivered in two modes: (1) same current to all locations (constant) or (2) current adjusted to perceptual threshold of each location (varied). For each stimulus location, subjects were asked to identify the location of the stimulus. Mechanical stimulation of the same locations on the fingerpad was delivered through von Frey hairs (0.07, 0.2 and 0.4 g). The percentage of accurate responses was computed for all stimulation modes. We found that the accuracy of discrimination of stimulus location in both the constant (46%) and varied (40%) electrotactile stimulation modes was significantly higher than chance level (25%; p < 0.01). Furthermore, subjects were significantly more accurate in discriminating electrotactile stimuli in the constant than in the varied mode (p < 0.05). We also found that the accuracy of spatial discrimination was dependent on stimulation site for mechanical, but not electrotactile stimulation. Finally, we found a significant difference in accuracy over the duration of the experiment only for mechanical modes, which may indicate that electrotactile stimuli are less biased over time. These results suggest that, although low in accuracy, human subjects are able to extract spatial information from electrotactile stimuli. Further research is needed to optimize the amount of the information that can be delivered through electrotactile stimulation.

Malignant hypertension presenting as acute pancreatitis.

We report a patient who was admitted to hospital with acute pancreatitis but who also had malignant phase hypertension. Whilst his alcohol intake was high, there was no objective evidence of alcoholic liver disease and no other underlying cause for pancreatitis was found. The pancreatitis may therefore have been due to pancreatic infarctions associated with fibrinoid necrosis. In all patients with acute pancreatitis, the diagnosis of malignant hypertension should be considered.

Immobilized polyphosphate kinase: preparation, properties, and potential for use in adenosine 5'-triphosphate regeneration.

Polyphosphate kinase (ATP:polyphosphate phosphotransferase; EC 2.7.4.1), partially purified from Escherichia coli, has been immobilized on glutaraldehyde-activated aminoethyl cellulose with a 10% retention of enzymatic activity. The immobilized enzyme can carry out the synthesis of ATP from ADP, using long-chain inorganic polyphosphate as a phosphoryl donor. Chromatographic analyses of the product mixture produced from ADP and [32P]polyphosphate demonstrated that 98% of the 32P was incorporated into ATP, indicating that the immobilized polyphosphate kinase is substantially free from contaminating polyphosphate phosphohydrolase (EC 3.6.1.11), adenosine triphosphatase (EC 3.6.1.4), and adenylate kinase (EC 2.7.4.3). Immobilized polyphosphate kinase loses no activity when stored in an aqueous suspension for 2 months at 5 degrees C or for 1-2 weeks at 25 degrees C. It may be stored indefinitely as a lyophilized powder at -10 degrees C. Michaelis constants for ADP and polyphosphate were determined to be 160 and 120 microM, respectively, for the immobilized enzyme. A small-batch reactor was found to produce ATP linearly with time up to 65% conversion of polyphosphate into ATP and to attain greater than 85% conversion to ATP at equilibrium. The ease of purification and immobilization of E. coli polyphosphate kinase, its storage stability, the purity and yield of its ATP product, and the low values of the Michaelis constants for its substrates make it a highly promising enzyme for ATP regeneration.