Evidence for surfactant contributing to the gastric mucosal barrier of the horse.

This study was undertaken to determine the hydrophobicity of the luminal surface of the equine stomach and to elucidate the ultrastructure of the lining imparting that property. Gastric and duodenal mucosal samples from 5 horses were collected immediately after euthanasia and subjected to surface contact angle measurement using a goniometer. Gastric mucosal samples from 4 horses and a foal were examined by electron microscopy following a fixation procedure known to preserve phospholipids and oligolamellar structures. Contact angles for the equine gastric glandular mucosal surface (mean +/- s.e. 78.0 +/- 11.0 degrees) were greater than for the duodenum (33.4 +/- 8.7 degrees), (P = 0.003). The contact angles for gastric squamous tissue (50.4 +/- 4.5 degrees) tended to be greater than for duodenum (P = 0.15). Electron microscopy revealed the existence of surfactant as abundant osmiophilic phospholipid material within both squamous and glandular gastric mucosae. These results indicate the hydrophobic nature of the equine gastric mucosae. We propose that the water-repellent nature of the stomach contributes to the 'gastric mucosal barrier' and is imparted by surface-active phospholipid adsorbed to the surface. Phospholipids may also be utilised as a physical barrier to back-diffusion of acid by lining intracellular canaliculi and oxyntic ducts where other defence mechanisms are absent.

In vitro elution of gentamicin, amikacin, and ceftiofur from polymethylmethacrylate and hydroxyapatite cement.

OBJECTIVE: To compare the elution characteristics of ceftiofur and liquid and powdered gentamicin and amikacin from polymethylmethacrylate (PMMA) and from hydroxyapatite cement (HAC). METHODS: PMMA and HAC beads in triplicate were impregnated with various amounts and formulations of antibiotics. Beads were immersed in 5 mL of phosphate buffered saline that was replaced at 1, 3, 6, and 12 hours, and 1, 2, 3, 5, 7, 10, 14, 18, 22, 26, and 30 days. The eluent was stored at -70 degrees C until assayed within 2 weeks by microbiological assay (gentamicin and amikacin) or capillary electrophoresis (ceftiofur). RESULTS: Rate of elution for all beads was greatest within the first 24 hours. Cumulative release of total antibiotic dose from beads over 30 days was significantly greater from HAC than PMMA. Antibiotic elution was directly related to the amount of antibiotic incorporated into the cement. Powdered and liquid forms of gentamicin had similar elution rates from PMMA. Elution of amikacin from PMMA beads was greater when the powdered form was used compared with liquid amikacin. Eluent concentrations of ceftiofur were similar to those of the aminoglycosides during the first 3 to 7 days but then decreased precipitously by comparison. CONCLUSIONS: Elution of antibiotics from HAC was greater than from PMMA. Gentamicin- and amikacin-impregnated PMMA and HAC released bactericidal concentrations of antibiotic for at least 30 days. Ceftiofur-impregnated PMMA or HAC is unlikely to provide long-term bactericidal concentrations. CLINICAL RELEVANCE: Gentamicin and amikacin elute effectively from PMMA and HAC.

Modes of local drug delivery to the musculoskeletal system.

A number of methods for the local delivery of drugs to musculoskeletal tissues in the horse are now available. Further research is required to document the disposition of drugs delivered by such methods and to correlate this information with efficacy. Perhaps the greatest potential area for the methods discussed is the treatment of synovial and bone infections. To be able to provide high and sustained therapeutic concentrations of antimicrobials to the site of infection should increase the chances of success in such cases. These methods of drug delivery need to be used in conjunction with other management procedures, however, including bacterial culture and sensitivity procedures, systemic antimicrobials, surgical drainage, removal of dead bone or surgical implants, establishment of fracture stability, use of autogenous bone grafts, systemic NSAIDs, and rest.

Ultrastructure of the hydrophobic gastric surfactant barrier in the dog.

OBJECTIVES: To confirm the hydrophobicity of the luminal surface of the canine stomach and to elucidate the ultrastructure of the lining imparting that property. DESIGN AND PROCEDURES: Gastric and duodenal mucosal samples from eight dogs were collected immediately after euthanasia and subjected to contact angle measurement using a goniometer. Other samples were examined by electron microscopy following a fixation procedure known to preserve phospholipids and oligolamellar structures. RESULTS: Contact angles for the canine gastric mucosal surface (85.1 +/- 5.5) were significantly greater (P < 0.0001) than for the duodenum (24.0 +/- 1.7). Electron microscopy revealed the existence of surfactant as abundant osmiophilic phospholipid material within the gastric and duodenal mucosae. CONCLUSION: We have confirmed the hydrophobic nature of the canine gastric mucosa whereas the luminal surface of the duodenum is hydrophilic. We propose that the water-repellent nature of the canine gastric lining contributes to the 'gastric mucosal barrier' and is imparted by an oligolamellar layer of surface-active phospholipid ('gastric surfactant') adsorbed to the surface. Both gastric and duodenal mucosae may also utilise phospholipids as an intercellular defense mechanism in the event that tight junctions are breached by acid. It is tempting to speculate that a deficiency of gastric phospholipids may predispose dogs to ulcers. Further, exogenous administration of phospholipids may be useful in preventing gastric ulceration.

The synovial response to exogenous phospholipid (synovial surfactant) injected into the equine radiocarpal joint compared with that to prilocaine, hyaluronan and propylene glycol.

AIMS: To determine the effects of the intra-articular injection of surface-active phospholipid in a propylene glycol carrier on synovial fluid composition and joint function of horses, and to compare these effects with those observed after the intra-articular administration of prilocaine, hyaluronan and propylene glycol alone. METHODS: Twenty-four horses were randomly allocated to four treatment groups: Group 1 100 mg of surface-active phospholipid in 1 ml of propylene glycol; Group 2 1 ml of propylene glycol; Group 3 10 ml of prilocaine; Group 4 2 ml of hyaluronan. Left radiocarpal joints were injected with the treatments and the right radiocarpal joints were injected with volume-matched saline as controls. Examinations for lameness, arthrocenteses and synovial fluid analyses were performed before and at 1, 3 and 7 days after injection. RESULTS: No horses became lame but treated joints temporarily developed mild to moderate effusions. Synovial fluid analyses indicated significantly greater inflammation in treated compared to control joints and this difference was greatest 24 hours after injection. There were no differences between the four treatments based on synovial fluid analysis except for neutrophil counts and alkaline phosphatase activities, which were significantly higher in prilocaine-treated joints. CONCLUSION: In horses, the intra-articular injection of surface-active phospholipid in a propylene glycol carrier induces clinically insignificant, temporary abnormalities in synovial fluid. Surface-active phospholipid was no more injurious to the synovium than prilocaine or hyaluronan. None of the agents used in this experiment caused lameness when injected into the joints of horses. RELEVANCE: This dose and formulation appear suitable for use in future experiments investigating the efficacy of surface-active phospholipid in the treatment or prevention of osteoarthritis in horses.

Release of lubricating synovial surfactant by intra-articular steroid.

This study was undertaken to determine whether glucocorticosteroids promote the secretion of lubricating surfactant, i.e. surface-active phospholipid (SAPL), into the joint. A standard clinical dose (100 mg) of methylprednisolone acetate (MPA) in 2.5 ml of saline was injected into the load-bearing right radiocarpal joint of five horses and 2.5 ml of saline injected into each of the contralateral joints used as controls. Synovial fluid (SF) was aspirated from all 10 joints before injection and at intervals of 16 and 32 h after injection, and then analysed by standard methods. All test joints showed an elevated level of SAPL, the increases averaging 112% after 16 h and 76% after 32 h, which were highly significant relative to the control joints. A large increase at 16 h was also found in proteolipid as a possible further marker of surfactant release. Significant quantities of proteolipid were also found in human SF. Since intra-articular steroids can dramatically improve joint mobility in both humans and horses, it is proposed that part of the benefit may be derived from improved lubrication arising from the remarkable ability of SAPL to lubricate under high load. Other possible benefits of elevating surfactant levels in the joints include control of cartilage hydration, promotion of macrophage activity and the ability to scavenge oxygen free radicals.