Using microwave thermal ablation to develop a subtotal, cortical-sparing approach to the management of primary aldosteronism.

Objective: To investigate the feasibility and efficacy of localized, subtotal, cortical-sparing microwave thermal ablation (MTA) as a potential curative management for primary aldosteronism. The study investigated with equal importance the selected ablation of small volumes of adrenal cortex while sparing adjacent cortex. Method: An in-vivo study was carried out in swine (n = 8) where MTA was applied under direct visualization, to the adrenal glands at 45 W or 70 W for 60 s, using a lateral, side-firing probe and a non-penetrative approach. Animals were survived for 48 h post-procedurally. Animals were investigated for markers of histological, immunohistochemical and biochemical evidence of adrenal function and adrenal damage by assessing samples drawn intra-operatively and at the time of euthanasia. Results: Selected MTA (70 W for 60 s) successfully ablated small adrenocortical volumes (∼0.8 cm3) characterized by coagulative necrosis and abnormal expression of functional markers (CYP11B1 and CYP17). Non-ablated, adjacent cortex was not affected and preserved normal expression of functional markers, without increased expression of markers of heat damage (HSP-70 and HMGB-1). Limited adrenal medullary damage was demonstrated histologically, clinically and biochemically. Conclusion: MTA offers potential as an efficient methodology for delivering targeted subtotal cortical-sparing adrenal ablation. Image-guided targeted MTA may also represent a safe future modality for curative management of PA, in the setting of both unilateral and bilateral disease.

Expression of pleiotrophin, an important regulator of cell migration, is inhibited in intestinal epithelial cells by treatment with non-steroidal anti-inflammatory drugs.

Non-steroidal anti-inflammatory drugs (NSAIDs) are among the most widely used drugs for the suppression of inflammation and pain. However, the analgesic properties of NSAIDs are also associated with significant negative side effects, most notably in the gastrointestinal (GI) tract. Increasingly, evidence indicates that the ulcerogenic properties of some NSAIDs are not exclusively the result of inhibition of cyclooxygenase isoforms in the GI tract, and other mechanisms, including inhibition of cell migration and epithelial restitution, are being explored. Recently, microarray analysis was used to identify potential novel targets of NSAID activity in intestinal epithelial cells. Treated cells exhibited significant reductions in the gene expression of pleiotrophin (PTN), a cytokine and growth factor known to participate in angiogenesis and bone growth. This report aimed to confirm the microarray results reported previously, and to measure protein expression of PTN in intestinal epithelial cells. Furthermore, we also examined the effects of exogenous PTN on cell migration in the presence and absence of either NSAIDs with variable ulcerogenic potential or PTN-specific siRNA. Our results demonstrated that indomethacin and NS-398, two NSAIDs with ulcerogenic potential significantly decrease both gene and protein expressions of PTN in IEC-6 cells and protein expression in IEC-6-Cdx2 cells. Additionally, cell migration experiments with PTN siRNA showed that PTN is an important mediator of IEC-6 cell migration, and addition of exogenous PTN partially restores the deficits in cell migration caused by treatment with indomethacin and NS-398. Finally, measurement of PTN protein expression in the GI tract of horses treated with phenylbutazone showed that PTN expression is reduced by NSAIDs in vivo. Our results show that PTN is an important mediator of cell migration in IEC-6 cells, and PTN is a potential target through which NSAIDs may inhibit cell migration, epithelial restitution, and wound healing in the GI tract.

Effect of a collateral ligament sparing surgical approach on mechanical properties of equine proximal interphalangeal joint arthrodesis constructs.

OBJECTIVE: To (1) compare the effect of a collateral ligament sparing surgical approach with an open surgical approach on mechanical properties of proximal interphalangeal joint (PIPJ) arthrodesis, and (2) to determine the percentage of articular cartilage surface removed by transarticular (TA) drilling with different diameter drill bits. STUDY DESIGN: Randomized paired limb design. SAMPLE POPULATION: Cadaveric equine limbs (n=76). METHODS: Cadaveric PIPJ were drilled using a 3.5, 4.5, or 5.5 mm drill bit at 80-84° to the dorsal plane to remove articular cartilage and subchondral bone from the distal articular surface of the proximal phalanx (P1) and the proximal articular surface of the middle phalanx (P2). Bone ends were photographed and the percentage of the projected surface area that was denuded of cartilage was measured. PIPJ arthrodesis constructs (3-hole dynamic compression plate [DCP], two 5.5 mm TA screws inserted in lag fashion, medial and lateral to the DCP; DCP-TA) were created using 2 surgical approaches in paired limbs. A conventional open approach was used in 1 limb and a collateral ligament sparing approach used in the other limb. Constructs were tested to failure in single-cycle 3-point dorsopalmar/plantar or lateromedial bending. Maximum load, yield load, and composite stiffness were compared between techniques. RESULTS: The 3.5, 4.5, and 5.5 mm drill bits removed 24±4%, 35±5%, and 45±7% of total PIPJ articular cartilage surface, respectively. Constructs with the collateral ligament sparing approach had significantly greater mean yield load (11.3±2.8 versus 7.68±1.1 kN, P=.008) and mean maximum load (13.5±3.1 versus 10.1±1.94 kN, P=.02) under lateromedial bending. Under dorsopalmar/plantar bending there was no significant difference between surgical approaches. The collateral ligament sparing arthrodesis technique had a shorter surgical time (19±3 minutes) compared with the open technique (31±3 minutes). CONCLUSION: A collateral ligament sparing surgical approach to the PIPJ with removal of articular cartilage by TA drilling and arthrodesis by DCP-TA was faster and stronger in mediolateral bending than arthrodesis constructs created with an open surgical approach. CLINICAL RELEVANCE: Preservation of the collateral ligaments and TA drilling for cartilage removal during PIPJ arthrodesis may be a superior approach to the conventional open approach and warrants clinical evaluation.

Porcine vas deferens luminal pH is acutely increased by systemic xylazine administration.

Data are accumulating to demonstrate that pH regulation in the male reproductive tract has a vital role in modulating sperm cell fertilizing capacity, and therefore male fertility. Bicarbonate uptake by sperm cells is required for the achievement of motility levels required for fertilization. Vas deferens epithelial cells can carry out measurable bicarbonate secretion, but the available literature to date reports that the vas deferens luminal content is typically acidic. This study aimed to determine pH in the boar vas deferens lumen and whether modulatory mechanisms exist for regulation of pH in this compartment of the male reproductive tract. A fiberoptic pH probe was used to assess pH in the vas deferens of anesthetized adult boars. The mean pH, derived from multiple measurements at variable positions along the vas deferens lumen, was 7.39 +/- 0.09. Furthermore, administration of xylazine, an alpha-2 adrenergic receptor agonist rapidly (<10 min) alkalinized the vas deferens lumen in most cases. Because the duct was transected proximal to the site of measurements, the observations rule out the possibility that alkalinization resulted from secretion in more proximal portions of the duct. These results indicate that the boar vas deferens lumen can be alkaline, and they suggest that porcine vas deferens epithelia increase net bicarbonate secretion in vivo after systemic alpha-2 adrenergic stimulation. This secretory response greatly changes the luminal environment to which sperm cells are exposed, which will initiate or enhance motility, and is expected to modulate male fertility.

Isolation and Culture of Juvenile Pig Thyroid Follicular Epithelia.

Epithelial tissues are defined by their polarity and their ability to transport directionally. Thyroid is a tissue comprising functional epithelial units organized as enclosed follicles, with their luminal spaces defined by thyrocyte apices. Thus, the native arrangement of thyroid epithelia limits accessibility to the follicular space, presenting a challenge in studying transepithelial movements. This limitation can be overcome by studying thyrocytes grown as two-dimensional cultures. Herein we present methods for isolation of thyroid follicles from juvenile pigs and preparation of high-resistance, polarized cultures.

Suppression of calpain expression by NSAIDs is associated with inhibition of cell migration in rat duodenum.

Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used for the alleviation of pain and inflammation, but these drugs are also associated with a suite of negative side effects. Gastrointestinal (GI) toxicity is particularly concerning since it affects an estimated 70% of individuals taking NSAIDs routinely, and evidence suggests the majority of toxicity is occurring in the small intestine. Traditionally, NSAID-induced GI toxicity has been associated with indiscriminate inhibition of cyclooxygenase isoforms, but other mechanisms, including inhibition of cell migration, intestinal restitution, and wound healing, are likely to contribute to toxicity. Previous efforts demonstrated that treatment of cultured intestinal epithelial cells (IEC) with NSAIDs inhibits expression and activity of calpain proteases, but the effects of specific inhibition of calpain expression in vitro or the effects of NSAIDs on intestinal cell migration in vivo remain to be determined. Accordingly, we examined the effect of suppression of calpain protease expression with siRNA on cell migration in cultured IECs and evaluated the effects of NSAID treatment on epithelial cell migration and calpain protease expression in rat duodenum. Our results show that calpain siRNA inhibits protease expression and slows migration in cultured IECs. Additionally, NSAID treatment of rats slowed migration up the villus axis and suppressed calpain expression in duodenal epithelial cells. Our results are supportive of the hypothesis that suppression of calpain expression leading to slowing of cell migration is a potential mechanism through which NSAIDs cause GI toxicity.

Inhibition of Kv channel expression by NSAIDs depolarizes membrane potential and inhibits cell migration by disrupting calpain signaling.

Clinical use of non-steroidal anti-inflammatory drugs (NSAIDs) is well known to cause gastrointestinal ulcer formation via several mechanisms that include inhibiting epithelial cell migration and mucosal restitution. The drug-affected signaling pathways that contribute to inhibition of migration by NSAIDs are poorly understood, though previous studies have shown that NSAIDs depolarize membrane potential and suppress expression of calpain proteases and voltage-gated potassium (Kv) channel subunits. Kv channels play significant roles in cell migration and are targets of NSAID activity in white blood cells, but the specific functional effects of NSAID-induced changes in Kv channel expression, particularly on cell migration, are unknown in intestinal epithelial cells. Accordingly, we investigated the effects of NSAIDs on expression of Kv1.3, 1.4, and 1.6 in vitro and/or in vivo and evaluated the functional significance of loss of Kv subunit expression. Indomethacin or NS-398 reduced total and plasma membrane protein expression of Kv1.3 in cultured intestinal epithelial cells (IEC-6). Additionally, depolarization of membrane potential with margatoxin (MgTx), 40mM K(+), or silencing of Kv channel expression with siRNA significantly reduced IEC-6 cell migration and disrupted calpain activity. Furthermore, in rat small intestinal epithelia, indomethacin and NS-398 had significant, yet distinct, effects on gene and protein expression of Kv1.3, 1.4, or 1.6, suggesting that these may be clinically relevant targets. Our results show that inhibition of epithelial cell migration by NSAIDs is associated with decreased expression of Kv channel subunits, and provide a mechanism through which NSAIDs inhibit cell migration and may contribute to NSAID-induced gastrointestinal (GI) toxicity.

Structural and functional basis for the long QT syndrome: relevance to veterinary patients.

Long QT syndrome (LQTS) is a condition characterized by prolongation of ventricular repolarization and is manifested clinically by lengthening of the QT interval on the surface ECG. Whereas inherited forms of LQTS associated with mutations in the genes that encode ion channel proteins are identified only in humans, the acquired form of LQTS occurs in humans and companion animal species. Often, acquired LQTS is associated with drug-induced block of the cardiac K+ current designated I(Kr). However, not all drugs that induce potentially fatal ventricular arrhythmias antagonize I(Kr), and not all drugs that block I(Kr), are associated with ventricular arrhythmias. In clinical practice, the extent of QT interval prolongation and risk of ventricular arrhythmia associated with antagonism of I(Kr) are modulated by pharmacokinetic and pharmacodynamic variables. Veterinarians can influence some of the potential risk factors (eg, drug dosage, route of drug administration, presence or absence of concurrent drug therapy, and patient electrolyte status) but not all (eg, patient gender/genetic background). Veterinarians need to be aware of the potential for acquired LQTS during therapy with drugs identified as blockers of HERG channels and I(Kr).

Effects of immobilization followed by remobilization on mineral density, histomorphometric features, and formation of the bones of the metacarpophalangeal joint in horses.

OBJECTIVE: To determine microradiographic appearance, bone histomorphometry, and mineral density of the long bones of the metacarpophalangeal joint in horses after immobilization followed by remobilization. ANIMALS: 5 healthy horses. PROCEDURE: One forelimb of each horse was immobilized in a fiberglass cast for 7 weeks, followed by 8 weeks of increasing exercise. Calcein and oxytetracycline were administered IV during the immobilization and exercise phases, respectively, for bone labeling and analysis after euthanasia. Sagittal sections of metacarpal bones and proximal phalanges were examined via radiography, dual energy x-ray absorptiometry, histomorphometry, and bone label analysis. RESULTS: Radiography revealed loss of bone mineral opacity in the subarticular regions of the immobilized metacarpal bones and phalanges and subchondral lesions in metacarpal bones in 2 horses. In phalanges, a significant decrease in subarticular volumetric bone mineral density was detected. There was significantly less bone volume and calcein-labeled bone surface and more vascular volume and oxytetracycline-labeled bone surface in immobilized phalanges, compared with contralateral phalanges. CONCLUSIONS AND CLINICAL RELEVANCE: Eight weeks of exercise after single-limb immobilization is insufficient for recovery of volumetric bone mineral density. During immobilization and remobilization, the subchondral and trabecular bone appear to be actively remodeling.

Clinical evaluation of the effects of immobilization followed by remobilization and exercise on the metacarpophalangeal joint in horses.

OBJECTIVES: To evaluate clinical effects of immobilization followed by remobilization and exercise on the metacarpophalangeal joint (MPJ) in horses. ANIMALS: 5 healthy horses. PROCEDURE: After lameness, radiographic, and force plate examinations to determine musculoskeletal health, 1 forelimb of each horse was immobilized in a fiberglass cast for 7 weeks, followed by cast removal and increasing amounts of exercise, beginning with hand-walking and ending with treadmill exercise. Lameness examination, arthrocentesis of both MPJ, single-emulsion radiographic examination, nuclear scintigraphic examination, ground-reaction force-plate analysis, and computed tomographic examination were done at various times during the study. RESULTS: All horses were lame in the immobilized MPJ after cast removal; lameness improved slightly with exercise. Force plate analysis revealed a significant difference in peak forces between immobilized and contralateral limbs 2 weeks after cast removal. Range of motion of the immobilized MPJ was significantly decreased, and joint circumference was significantly increased, compared with baseline values, during the exercise period. Osteopenia was subjectively detected in the immobilized limbs. Significant increase in the uptake of radionucleotide within bones of the immobilized MPJ after cast removal and at the end of the study were detected. Loss of mineral opacity, increased vascular channels in the subchondral bone, and thickening within the soft tissues of the immobilized MPJ were detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that 8 weeks of enforced exercise after 7 weeks of joint immobilization did not restore joint function or values for various joint measurements determined prior to immobilization.

Expression of the ether-a-go-go (ERG) potassium channel in smooth muscle of the equine gastrointestinal tract and influence on activity of jejunal smooth muscle.

OBJECTIVE: To determine whether ether-a-go-go (ERG) potassium channels are expressed in equine gastrointestinal smooth muscle, whether ERG channel antagonists affect jejunal muscle contraction in vitro, and whether plasma cisapride concentrations in horses administered treatment for postoperative ileus (POI) are consistent with ERG channels as drug targets. SAMPLE POPULATION: Samples of intestinal smooth muscle obtained from 8 horses free of gastrointestinal tract disease and plasma samples obtained from 3 horses administered cisapride for treatment of POI. PROCEDURE: Membranes were prepared from the seromuscular layer of the duodenum, jejunum, ileum, cecum, large colon, and small colon. Immunoblotting was used to identify the ERG channel protein. Isolated jejunal muscle strips were used for isometric stress response to ERG channel blockers that included E-4031, MK-499, clofilium, and cisapride. Plasma concentrations of cisapride were determined in 3 horses administered cisapride for treatment of POI after small intestinal surgery. RESULTS: Immunoblotting identified ERG protein in all analyzed segments of the intestinal tract in all horses. The selective ERG antagonist E-4031 caused a concentration-dependent increase in jejunal contraction. Clofilium, MK-499, and cisapride also increased jejunal contraction at concentrations consistent with ERG channel block; effects of E-4031 and cisapride were not additive. Peak plasma cisapride concentrations in treated horses were consistent with ERG block as a mechanism of drug action. CONCLUSIONS AND CLINICAL RELEVANCE: The ERG potassium channels modulate motility of intestinal muscles in horses and may be a target for drugs. This finding may influence development of new prokinetic agents and impact treatment of horses with POI.

Segmental ostectomy of the second and fourth metacarpal and metatarsal bones in horses: 17 cases (1993-2002).

OBJECTIVE: To determine clinical findings in and outcome of horses with fractures of the second or fourth metacarpal or metatarsal bone that underwent segmental ostectomy, leaving the proximal and distal portions of the bone undisturbed. DESIGN: Retrospective case series. ANIMALS: 17 horses. PROCEDURES: Medical records were reviewed, and information on signalment, affected bone, lesion type, surgical procedure, amount of bone removed, and surgical and postsurgical complications was obtained. Follow-up information was obtained through telephone conversations with owners, trainers, and referring veterinarians. RESULTS: One horse had a fracture involving the distal third of the second metacarpal bone; 13 had fractures involving the middle third of the second metacarpal bone (n = 4), fourth metacarpal bone (5), or fourth metatarsal bone (4); and 3 had fractures involving the proximal third of the second (2) or fourth (1) metacarpal bone. Affected portions of the bones were surgically resected, leaving the proximal and distal portions undisturbed. All horses returned to previous performance levels without evidence of lameness. Cosmetic results were good to excellent. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that horses with a complicated injury of the proximal, middle, or distal portion of the second or fourth metacarpal or metatarsal bone may be successfully treated by means of segmental ostectomy of the abnormal portion of the bone.

Tenoscopic-assisted treatment of open digital flexor tendon sheath injuries in horses: 20 cases (1992-2001).

OBJECTIVE: To determine history, physical and diagnostic examination findings, medical treatment, and outcome of horses with open injuries to the digital flexor tendon sheath treated with the assistance of tenoscopy. DESIGN: Betrospective study. ANIMALS: 20 horses. PROCEDURE: Medical records of 20 horses with open injuries to the digital flexor tendon sheath were reviewed. Signalment, history, physical and diagnostic examination results, bacteriologic culture and susceptibility testing results, surgical and medical treatments, and follow-up examination results were determined. Outcome was determined by use of telephone interview or physical examination. RESULTS: All horses were treated with tenoscopic-assisted lavage and debridement. Eighteen horses survived, and 2 were euthanatized during treatment. All horses were either grade-4 or grade-5 lame before treatment. Ten horses returned to previous use. Four horses were considered mildly lame and in athletic use. Three horses were considered mechanically lame and are in use with reduced expectations. One horse was lost to follow-up after being sold. One horse was euthanatized for financial reasons and 1 because of complications from regional sepsis. CONCLUSIONS AND CLINICAL RELEVANCE: Tenoscopy appears to be a useful modality in the treatment of open injury to the digital flexor tendon sheath in horses. Direct viewing, guided debridement, and targeted large-volume lavage are advantages obtained with intrathecal arthroscopy. Tenoscopy, when combined with antimicrobial and anti-inflammatory treatment, appears to offer a good chance of survival for affected horses.

Azoospermia associated with bilateral segmental aplasia of the ductus deferens in a stallion.

A 3-year-old Quarter Horse halter stallion was referred for routine semen evaluation. Physically, the stallion's reproductive organs appeared normal. Repeated semen evaluations did not reveal any spermatozoa. Because high activities of alkaline phosphatase are detected in the epididymal fluid and indicative of complete ejaculation, alkaline phosphatase activities were analyzed in several samples, which yielded activities far less than reference values and suggested a blockage of the reproductive tract. Endoscopic evaluation of the urethra and the bulbourethral, prostate, and urethral gland ducts did not reveal abnormalities. The left ductus deferens was exposed surgically, and attempts to pass a catheter through it in a normograde direction met resistance after 20 cm. Laparoscopic abdominal surgery revealed the ductus deferens tapered to a thin structure just cranial to the entrance in the urogenital fold, cranial and lateral to the bladder. Both ductus deferentia were similarly affected. The symmetry and bilateral nature of the abnormalities were strong indications of a possible congenital defect.

In vivo assessment of a multicomponent and nanostructural polymeric matrix as a delivery system for antimicrobials and bone morphogenetic protein-2 in a unicortical tibial defect in goats.

OBJECTIVE: To determine the response of cortical bone to a multicomponent and nanostructural polymeric matrix as a drug delivery system for enhancing bone healing. ANIMALS: 20 healthy adult crossbred goats. PROCEDURES: A 3.5-mm-diameter unicortical defect was created in each tibia (day 0), and goats (4 goats/group) were treated as follows: not treated (control group), grafted with the matrix, grafted with antimicrobial (tigecycline and tobramycin)-impregnated matrix, grafted with recombinant human bone morphogenetic protein type 2 (rhBMP-2)-impregnated matrix, or grafted with antimicrobial- and rhBMP-2-impregnated matrix. Elution kinetics of antimicrobials was monitored through plasma concentrations. Bone response was assessed with radiographic scoring (days 1 and 30) and dual-energy x-ray absorptiometry (days 1, 14, and 30). Goats were euthanized on day 30, and histomorphologic analysis was performed. Categorical variables were analyzed with a generalized linear model, and continuous variables were analyzed with an ANOVA. RESULTS: Plasma antimicrobial concentrations indicated continued release throughout the study. Radiography and dual-energy x-ray absorptiometry did not reveal significant differences among treatments on day 30. Periosteal reactions were significantly greater surrounding bone defects grafted with rhBMP-2-impregnated matrix than those not treated or grafted with matrix or with antimicrobial-impregnated matrix; periosteal reactions were similar in bone defects grafted with rhBMP-2-impregnated matrix and antimicrobial- and rhBMP-2-impregnated matrix. CONCLUSIONS AND CLINICAL RELEVANCE: The matrix served as an antimicrobial delivery system and stimulated bone proliferation when rhBMP-2 was present. Antimicrobial and rhBMP-2 can be used concurrently, but the presence of antimicrobials may affect the performance of rhBMP-2.

Intra-abdominal hyaluronan concentration in peritoneal fluid of horses with sudden signs of severe abdominal pain.

OBJECTIVE: To determine hyaluronan concentrations in peritoneal fluid from healthy horses and horses with sudden signs of severe abdominal pain and to identify the cellular sources of hyaluronan within the peritoneal cavity. ANIMALS: 7 client-owned horses that were evaluated for sudden signs of severe abdominal pain, 6 healthy teaching horses, and 13 euthanized horses (11 with no abdominal disease and 2 that had undergone abdominal surgery 2 weeks previously for a different study). PROCEDURES: Abdominal fluid was collected from the client-owned and teaching horses. Hyaluronan concentrations were determined with an ELISA. Equine mesothelial cells were aseptically harvested from euthanized horses immediately after euthanasia, cultured, and processed for western blot immunoassays to detect expression of the following mesothelial cell markers: cytokeratins 8 and 18, vimentin, calretinin, mesothelin, and CD44. A reverse transcriptase-PCR assay was used to detect genetic expression of hyaluronan synthase-2 (HAS-2) from cultured and native equine tissue. RESULTS: The mean ± SD abdominal hyaluronan concentration in peritoneal fluid from horses with signs of abdominal pain (1,203.3 ± 46.3 ng/mL) was significantly greater than that in healthy horses (228.4 ± 167.3 ng/mL). Harvested cells were maintained, and immunoblotting analyses confirmed expression of the mesothelial markers. Gene expression of HAS-2 from cultured mesothelial cells and fibroblasts was confirmed. CONCLUSIONS AND CLINICAL RELEVANCE: Peritoneal hyaluronan concentration was much higher in horses with severe abdominal pain than in healthy horses. Cultured equine mesothelial cells and fibroblasts can produce hyaluronan through HAS-2. Future investigation should focus on establishing the effect of exogenous hyaluronan administration on mesothelial cell function in horses with abdominal disease.

Non-steroidal anti-inflammatory drugs inhibit calpain activity and membrane localization of calpain 2 protease.

Non-steroidal anti-inflammatory drugs (NSAIDs) are used frequently worldwide for the alleviation of pain despite their capacity to cause adverse gastrointestinal (GI) side effects. GI toxicity, once thought to be the result of non-specific inhibition of cyclooxegenase (COX) enzymes, is now hypothesized to have multiple other causes that are COX independent. In particular, NSAIDs inhibit intestinal epithelial restitution, the process by which barrier function in intestinal mucosa is restored at sites of epithelial wounds within hours through cell spreading and migration. Accordingly, recent evidence indicates that the expression of calpain proteases, which play a key role in cell migration, is decreased by NSAIDs that inhibit cell migration in intestinal epithelial cells (IEC). Here, we examine the effect of NSAIDs on calpain activity and membrane expression in IEC-6 cells. Indomethacin, NS-398, and SC-560 inhibited calpain activity and decreased expression of calpain 2 in total membrane fractions and in plasma membranes involved in cell attachment to the substrate. Additionally, we demonstrated that inhibition of calpain activity by NSAIDs or ALLM, a calpain inhibitor, limits cell migration and in vitro wound healing of IEC-6 cells. Our results indicate that NSAIDs may inhibit cell migration by decreasing calpain activity and membrane-associated expression of calpain 2. Our results provide valuable insight into the mechanisms behind NSAID-induced GI toxicity and provide a potential pathway through which these negative side effects can be avoided in future members of the NSAID class.

Ex vivo strength comparison of bioabsorbable tendon plates and bioabsorbable suture in a 3-loop pulley pattern for repair of transected flexor tendons from horse cadavers.

OBJECTIVE: To test the failure strength and energy of 2 bioabsorbable implants applied to transected deep digital flexor tendons (DDFT) from adult horses. STUDY DESIGN: Ex vivo biomechanical experiment. SAMPLE POPULATION: Twelve pairs of deep digital flexor tendons harvested from the forelimbs of fresh equine cadavers. METHODS: Poly-L-lactic acid tendon plates were custom manufactured for application to the cylindrical surface of an adult equine deep digital flexor tendon. Twelve pairs of DDFTs were transected 2 cm distal to the insertion of the distal check ligament of the deep digital flexor tendon. One tendon of each pair was randomly selected for repair with a biodegradable plate or a 3-loop pulley method. Size 2 polydioxanone suture was used in both repairs. Repairs were tested in tension to failure, with peak force (PF) and total energy (TE) at repair failure recorded in Newtons (N) and Joules (J), respectively. A paired t-test was used for statistical evaluation with a significant level set at P< or = .05. RESULTS: Mean+/-SD PF for failure of plated tendons (1507.08+/-184.34 N) was significantly greater than for sutured tendons (460.86+/-60.93 N). TE was also significantly greater for failure of plated tendons versus sutured tendons. CONCLUSIONS: Plate fixation of transected cadaver DDFTs appear to have superior immediate failure strength than 3-loop pulley repairs. CLINICAL RELEVANCE: Whereas in vivo testing is required, a bioabsorbable tendon plate may provide initial increased strength to support tendon healing and decrease external coaptation requirements.