Acute- and late-phase matrix metalloproteinase (MMP)-9 activity is comparable in female and male rats after peripheral nerve injury.

BACKGROUND:In the peripheral nerve, pro-inflammatory matrix metalloproteinase (MMP)-9 performs essential functions in the acute response to injury. Whether MMP-9 activity contributes to late-phase injury or whether MMP-9 expression or activity after nerve injury is sexually dimorphic remains unknown. METHODS:Patterns of MMP-9 expression, activity and excretion were assessed in a model of painful peripheral neuropathy, sciatic nerve chronic constriction injury (CCI), in female and male rats. Real-time Taqman RT-PCR for MMP-9 and its endogenous inhibitor, tissue inhibitor of metalloproteinase-1 (TIMP-1) of nerve samples over a 2-month time course of CCI was followed by gelatin zymography of crude nerve extracts and purified MMP-9 from the extracts using gelatin Sepharose-beads. MMP excretion was determined using protease activity assay of urine in female and male rats with CCI. RESULTS:The initial upsurge in nerve MMP-9 expression at day 1 post-CCI was superseded more than 100-fold at day 28 post-CCI. The high level of MMP-9 expression in late-phase nerve injury was accompanied by the reduction in TIMP-1 level. The absence of MMP-9 in the normal nerve and the presence of multiple MMP-9 species (the proenzyme, mature enzyme, homodimers, and heterodimers) was observed at day 1 and day 28 post-CCI. The MMP-9 proenzyme and mature enzyme species dominated in the early- and late-phase nerve injury, consistent with the high and low level of TIMP-1 expression, respectively. The elevated nerve MMP-9 levels corresponded to the elevated urinary MMP excretion post-CCI. All of these findings were comparable in female and male rodents. CONCLUSION:The present study offers the first evidence for the excessive, uninhibited proteolytic MMP-9 activity during late-phase painful peripheral neuropathy and suggests that the pattern of MMP-9 expression, activity, and excretion after peripheral nerve injury is universal in both sexes

The alternatively spliced fibronectin CS1 isoform regulates IL-17A levels and mechanical allodynia after peripheral nerve injury.

BackgroundMechanical pain hypersensitivity associated with physical trauma to peripheral nerve depends on T-helper (Th) cells expressing the algesic cytokine, interleukin (IL)-17A. Fibronectin (FN) isoform alternatively spliced within the IIICS region encoding the 25-residue-long connecting segment 1 (CS1) regulates T cell recruitment to the sites of inflammation. Herein, we analyzed the role of CS1-containing FN (FN-CS1) in IL-17A expression and pain after peripheral nerve damage.MethodsMass spectrometry, immunoblotting, and FN-CS1-specific immunofluorescence analyses were employed to examine FN expression after chronic constriction injury (CCI) in rat sciatic nerves. The acute intra-sciatic nerve injection of the synthetic CS1 peptide (a competitive inhibitor of the FN-CS1/α4 integrin binding) was used to elucidate the functional significance of FN-CS1 in mechanical and thermal pain hypersensitivity and IL-17A expression (by quantitative Taqman RT-PCR) after CCI. The CS1 peptide effects were analyzed in cultured primary Schwann cells, the major source of FN-CS1 in CCI nerves.ResultsFollowing CCI, FN expression in sciatic nerve increased with the dominant FN-CS1 deposition in endothelial cells, Schwann cells, and macrophages. Acute CS1 therapy attenuated mechanical allodynia (pain from innocuous stimulation) but not thermal hyperalgesia and reduced the levels of IL-17A expression in the injured nerve. CS1 peptide inhibited the LPS- or starvation-stimulated activation of the stress ERK/MAPK pathway in cultured Schwann cells.ConclusionsAfter physical trauma to the peripheral nerve, FN-CS1 contributes to mechanical pain hypersensitivity by increasing the number of IL-17A-expressing (presumably, Th17) cells. CS1 peptide therapy can be developed for pharmacological control of neuropathic pain

Evaluation of neurotoxicity and long-term function and behavior following intrathecal 1 % 2-chloroprocaine in juvenile rats

Spinally-administered local anesthetics provide effective perioperative anesthesia and/or analgesia for children of all ages. New preparations and drugs require preclinical safety testing in developmental models. We evaluated age-dependent efficacy and safety following 1 % preservative-free 2-chloroprocaine (2-CP) in juvenile Sprague-Dawley rats. Percutaneous lumbar intrathecal 2-CP was administered at postnatal day (P)7, 14 or 21. Mechanical withdrawal threshold pre- and post-injection evaluated the degree and duration of sensory block, compared to intrathecal saline and naive controls. Tissue analyses one- or seven-days following injection included histopathology of spinal cord, cauda equina and brain sections, and quantification of neuronal apoptosis and glial reactivity in lumbar spinal cord. Following intrathecal 2-CP or saline at P7, outcomes assessed between P30 and P72 included: spinal reflex sensitivity (hindlimb thermal latency, mechanical threshold); social approach (novel rat versus object); locomotor activity and anxiety (open field with brightly-lit center); exploratory behavior (rearings, holepoking); sensorimotor gating (acoustic startle, prepulse inhibition); and learning (Morris Water Maze). Maximum tolerated doses of intrathecal 2-CP varied with age (1.0 μL/g at P7, 0.75 μL/g at P14, 0.5 μL/g at P21) and produced motor and sensory block for 10−15 min. Tissue analyses found no significant differences across intrathecal 2-CP, saline or naïve groups. Adult behavioral measures showed expected sex-dependent differences, that did not differ between 2-CP and saline groups. Single maximum tolerated in vivo doses of intrathecal 2-CP produced reversible spinal anesthesia in juvenile rodents without detectable evidence of developmental neurotoxicity. Current results cannot be extrapolated to repeated dosing or prolonged infusion

Quantification of left ventricular remodeling in response to isolated aortic or mitral regurgitation

<p>Abstract</p> <p>Background</p> <p>The treatment of patients with aortic regurgitation (AR) or mitral regurgitation (MR) relies on the accurate assessment of the severity of the regurgitation as well as its effect on left ventricular (LV) size and function. Cardiovascular Magnetic Resonance (CMR) is an excellent tool for quantifying regurgitant volumes as well as LV size and function. The 2008 AHA/ACC management guidelines for the therapy of patients with AR or MR only describe LV size in terms of linear dimensions (i.e. end-diastolic and end-systolic dimension). LV volumes that correspond to these linear dimensions have not been published in the peer-reviewed literature. The purpose of this study is to determine the effect of regurgitant volume on LV volumes and chamber dimensions in patients with isolated AR or MR and preserved LV function.</p> <p>Methods</p> <p>Regurgitant volume, LV volume, mass, linear dimensions, and ejection fraction, were determined in 34 consecutive patients with isolated AR and 23 consecutive patients with MR and no other known cardiac disease.</p> <p>Results</p> <p>There is a strong, linear relationship between regurgitant volume and LV end-diastolic volume index (aortic regurgitation r²= 0.8, mitral regurgitation r²= 0.8). Bland-Altman analysis of regurgitant volume shows little interobserver variation (AR: 0.6 ± 4 ml; MR 4 ± 6 ml). The correlation is much poorer between regurgitant volume and commonly used clinical linear measures such as end-systolic dimension (mitral regurgitation r²= 0.3, aortic regurgitation r²= 0.5). For a given regurgitant volume, AR causes greater LV enlargement and hypertrophy than MR.</p> <p>Conclusion</p> <p>CMR is an accurate and robust technique for quantifying regurgitant volume in patients with AR or MR. Ventricular volumes show a stronger correlation with regurgitant volume than linear dimensions, suggesting LV volumes better reflect ventricular remodeling in patients with isolated mitral or aortic regurgitation. Ventricular volumes that correspond to published recommended linear dimensions are determined to guide the timing of surgical intervention.</p

Evaluation of Spinal Toxicity and Long-term Spinal Reflex Function after Intrathecal Levobupivaciane in the Neonatal Rat

Neuraxial anesthesia is utilized in children of all ages. Local anesthetics produce dose-dependent toxicity in certain adult models, but the developing spinal cord may also be susceptible to drug-induced apoptosis. In postnatal rodents, we examined the effects of intrathecal levobupivacaine on neuropathology and long-term sensorimotor outcomes

Matrix metalloproteinases at key junctions in the pathomechanism of stroke

Matrix metalloproteinases play a crucial role in the remodelling of the extracellular matrix through direct degradation of its structural proteins and control of extracellular signaling. The most common cause of ischemic brain damage is an atherothrombotic lesion in the supplying arteries. The progress of the atherosclerotic plaque development and the related thrombotic complications are mediated in part by matrix metalloproteinases. In addition to their role in the underlying disease, various members of this protease family are upregulated in the acute phase of ischemic brain damage as well as in the post-ischemic brain recovery following stroke. This review summarizes the current understanding of the matrix metalloproteinase-related molecular events at three stages of the ischemic cerebrovascular disease (in the atherosclerotic plaque, in the neurovascular unit of the brain and in the regenerating brain tissue)

In Vivo Evaluation of Quantitative Percussion Diagnostics for Determining Implant Stability

PURPOSE: A percussion instrument (Periometer(®), Perimetrics LLC, Newport Beach, CA, USA) and rat model were used to test the hypothesis: percussion diagnostics provides reliable, reproducible indications of osseointegration. MATERIALS AND METHODS: Titanium implants were placed in femurs of 36 Sprague-Dawley rats. Each animal was assigned to one of six groups of six defined by one of three time points (2, 4, or 8 weeks post-placement) and one of two treatments (MMP inhibitor or vehicle). Percussion testing was conducted three times/subject at implant placement and at one of the time points. For each time point, there was an experimental group that received daily intraperitoneal injections of GM6001, and a control group that received no MMP inhibitor. The percussion data consisted of loss coefficient (LC) values that characterize energy dissipation. Statistical analysis was performed on the LC values for two animal groups using the paired Student t test to assess differences as a function of time, and the independent t test to compare mean LC for the study groups at sacrifice (α=0.05). Histological evaluation using the osteogenic CD40 protein marker was also performed. RESULTS: A nearly significant difference in mean LC at the 2-week time point was observed between the two treatments with the GM6001 group having the higher value (p = 0.053). There was a greater difference between the mean LC values for the 4-week GM6001 and vehicle groups (p = 0.001). The histological evidence for subjects in these two groups confirmed reduction of osteogenesis at the implant interface after administration of the MMP inhibitor. CONCLUSIONS: Lower vehicle LC values relative to the GM6001 therapeutic group were observed, consistent with the effect MMP inhibition has on matrix remodeling at the implant bone interface. This finding in conjunction with histological observations confirms that osseointegration can be monitored using percussion diagnostics