Considerations for the design and execution of protocols for animal research and treatment to improve reproducibility and standardization: "DEPART well-prepared and ARRIVE safely".

OBJECTIVE: To review the factors in experimental design that contribute to poor translation of pre-clinical research to therapies for patients with osteoarthritis (OA) and how this might be improved. METHODS: Narrative review of the literature, and evaluation of the different stages of design conduct and analysis of studies using animal models of OA to define specific issues that might reduce quality of evidence and how this can be minimised. RESULTS: Preventing bias and improving experimental rigour and reporting are important modifiable factors to improve translation from pre-clinical animal models to successful clinical trials of therapeutic agents. Despite publication and adoption by many journals of guidelines such as Animals in Research: Reporting In Vivo Experiments (ARRIVE), experimental animal studies published in leading rheumatology journals are still deficient in their reporting. In part, this may be caused by researchers first consulting these guidelines after the completion of experiments, at the time of publication. This review discusses factors that can (1) bias the outcome of experimental studies using animal models of osteoarthritis or (2) alter the quality of evidence for translation. We propose a checklist to consult prior to starting experiments; in the Design and Execution of Protocols for Animal Research and Treatment (DEPART). CONCLUSIONS: Following DEPART during the design phase will enable completion of the ARRIVE checklist at the time of publication, and thus improve the quality of evidence for inclusion of experimental animal research in meta-analyses and systematic reviews: "DEPART well-prepared and ARRIVE safely".

A non-invasive, 3D, dynamic MRI method for measuring muscle moment arms in vivo: demonstration in the human ankle joint and Achilles tendon.

Muscle moment arms are used widely in biomechanical analyses. Often they are measured in 2D or at a series of static joint positions. In the present study we demonstrate a simple MRI method for measuring muscle moment arms dynamically in 3D from a single range-of-motion cycle. We demonstrate this method in the Achilles tendon for comparison with other methods, and validate the method using a custom apparatus. The method involves registration of high-resolution joint geometry from MRI scans of the stationary joint with low-resolution geometries from ultrafast MRI scans of the slowly moving joint. Tibio-talar helical axes and 3D Achilles tendon moment arms were calculated throughout passive rotation for 10 adult subjects, and compared with recently published data. A simple validation was conducted by comparing MRI measurements with direct physical measurements made on a phantom. The moment arms measured using our method and those of others were similar and there was good agreement between physical measurements (mean 41.0mm) and MRI measurements (mean 39.5mm) made on the phantom. This new method can accurately measure muscle moment arms from a single range-of-motion cycle without the need to control rotation rate or gate the scanning. Supplementary data includes custom software to assist implementation.

Using static preload with magnetic resonance elastography to estimate large strain viscoelastic properties of bovine liver.

Traditional magnetic resonance elastography (MRE) applies small amplitude vibration to tissues. Thus currently MRE measures only the small deformation behaviour of tissues. MRE has the potential to estimate the strain-varying shear modulus of soft tissues, if applied at different static strains, which may allow prediction of the large-strain behaviour of tissues. This study uses MRE of bovine liver specimens under various levels of static compressive pre-strain up to 30%. Storage and loss moduli measured using MRE increased non-linearly with static compressive pre-strain, and exponential models fit well to these data to describe this relationship (R(2)>0.93). Based on these models, a 10% linear compression of liver would result in a 47% overestimate of the 'true' storage modulus of the uncompressed tissue. The results of this study have implications for MRE transducer design and interpretation of results from in vivo MRE studies.

A CCl4/CHCl3 interaction study in isolated hepatocytes: non-induced and phenobarbital-pretreated cells.

The purpose of this study was to evaluate an isolated hepatocyte model for predicting the in vivo hepatotoxicity of carbon tetrachloride (CCl4) and chloroform (CHCl3), alone and in combination. Response surface methodology (RSM) was used to analyze and describe the data. The interaction was evaluated for % initial K+ (cell injury) and % LDH leakage (cell death) in non-induced (untreated) and phenobarbital-pretreated suspended hepatocytes. CCl4 and CHCl3 were delivered alone and in combination in dimethyl sulfoxide (DMSO) to suspended hepatocytes. The maximum observed no-effect level (MONEL) for CCl4 in non-induced cells was 1.0 mM (LDH and K+). In induced cells, the MONEL was 0.25 mM (K+) and 0.5 mM (LDH). The MONEL for CHCl3 in non-induced cells was 5.0 mM (LDH and K+) and in induced cells was 0.5 mM (K+) and 1.0 mM (LDH). Phenobarbital pretreatment enhanced the toxicity of both CCl4 and CHCl3, alone and in combination. RSM analysis of the % initial K+ and % LDH for CCl4 and CHCl3 in combination in noninduced and induced cells showed a greater than additive interaction. The isolated hepatocyte model appears to be a promising system for evaluating the toxicity of chemical mixtures and predicting their in vivo effects.

A CCl4/CHCl3 interaction study in isolated hepatocytes: selection of a vehicle.

Emulphor, ethanol, and dimethyl sulfoxide (DMSO) were evaluated as vehicles in studying the toxicity of CCl4 and CHCl3 in isolated hepatocytes. The appropriateness of the vehicle was determined by evaluating the following parameters: solubility of CCl4 and CHCl3 in the vehicle, cell injury (intracellular K+), cell death (LDH leakage), and lack of interaction (protection or enhanced toxicity) with CCl4 and CHCl3. The relative toxicity of the vehicles according to maximum no effect levels (v/v) was: emulphor (0.125%) greater than ethanol (1.0%) greater than DMSO (5.0%). Emulphor at toxic levels was inadequate to dissolve enough CCl4 to evaluate in this system. Ethanol (5.0, 2.5, 1.0, 0.5%) was more toxic than DMSO and interacted with both CCl4 and CHCl3 to enhance toxicity. DMSO (15.0, 5.0, 2.5%) did not significantly alter the toxicity of CCl4 and CHCl3; no interaction. These data suggest that DMSO should be the vehicle for evaluating the toxicity of CCl4 and CHCl3 and their mechanisms of action in the isolated hepatocyte.

Clinical and immunological reactions to Aspergillus niger among workers at a biotechnology plant.

The workforce at a biotechnology plant producing citric acid by fermentation of molasses with a strain of Aspergillus niger was studied. A combination of a respiratory questionnaire and clinical assessment identified 18 subjects (4.9% of the workforce) with work related bronchospasm. In nine of these evidence of sensitisation to A niger was obtained by skin prick tests and radioallergosorbent test (RAST) using as an antigen an extract of the A niger culture fluid from the process. Of the 325 subjects without work related bronchospasm, only nine (2.7%) had a positive prick test. There were no subjects with symptoms of extrinsic allergic alveolitis. Investigation into the source of the antigen showed that whereas, in some areas of the plant, A niger spores were present, in others there were no detectable spores. In these areas, however, extracts of filters from air samplers were shown by RAST inhibition to contain A niger antigens, indicating that the culture fluid was generating airborne antigen. RAST inhibition studies showed that the A niger culture fluid used in the process contained antigens that were not present in a commercially available A niger extract, thus emphasising the importance in this type of investigation of using antigens prepared from material to which the workers are exposed.