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Abstract

Case Description—A 4-year-old horse was evaluated for chronic coughing, mucopurulent nasal discharge, and failure to thrive.

Clinical Findings—Endoscopy was performed on the larynx, pharynx, and trachea and revealed an approximately 2-cm fluctuant subepiglottic cyst.

Treatment and Outcome—The subepiglottic cyst was treated with intralesional formalin administration. Following 2 injections, 2 weeks apart, the cyst was completely resolved with no evidence of scarring or epiglottic deformity.

Clinical Relevance—Findings suggest that intralesional formalin administration for treatment of subepiglottic cysts may be a minimally invasive, economically suitable alternative to surgical treatments.

Full access
in Journal of the American Veterinary Medical Association

Abstract

Objective—To evaluate 2 commercially available transfection reagents for transfection efficiency and distribution of small interfering RNA (siRNA) molecules to chondrocytes in monolayer cultures and full-thickness cartilage explants from guinea pigs and horses.

Sample—Cartilage explants from 5 one-month-old and 3 adult guinea pigs and 5 adult clinically normal horses.

Procedures—Monolayer chondrocytes and uniform cartilage explants were exposed to 1 of 2 siRNA transfection complexes according to manufacturers' protocols (1μM [1×]). Additionally, monolayer chondrocytes were exposed to 2× the suggested amount of a proprietary siRNA molecule. Full-thickness cartilage explants were treated with 1× (1μM), 2× (2μM), and 4× (4μM) or 1× (0.13μM), 4× (0.52μM), and 8× (1.04μM) the recommended concentrations of the proprietary siRNA and the cationic liposome siRNA, respectively, in equivalent media volumes. Use of fluorescent siRNA duplexes allowed quantification of transfected cells via flow cytometry and direct visualization of the depth and distribution of in situ transfection via fluorescent microscopy.

Results—With both transfection reagents, > 90% of monolayer chondrocytes were transfected. In explants, only use of the proprietary molecule achieved > 50% transfection efficiency, whereas use of the cationic liposome achieved < 20%. Only the proprietary molecule-treated cartilage consistently contained fluorescent cells throughout all zones; the cationic liposome-transfected chondrocytes were restricted to explant surfaces.

Conclusions and Clinical Relevance—Robust transfection of chondrocytes in monolayer was achieved with both reagents, but only use of the proprietary molecule attained effective full-thickness transfection of explants that may allow relevant transcript reduction via RNAi.

Full access
in American Journal of Veterinary Research