RESUMO
PURPOSE: To develop a model for experimental Streptococcus pneumoniae keratitis and to evaluate the chemotherapeutic efficacy of 12 common topical antibiotics in vivo. METHODS: Five-hundred (CFUs of log-phase S. pneumoniae were injected into the central corneal stroma of 36 eyes of 18 rabbits. After 0, 4, 8, 16, 24, and 48 hours, the in vivo growth was assayed as the CFU per cornea. Epithelial removal (to promote antibiotic entry and mimic human keratitis) was evaluated. Disc or tube dilution verification of the sensitivity or resistance of three S. pneumoniae strains was performed: a penicillin sensitive ("S"), an intermediate sensitive ("I"), and a resistant ("R") strain. Keratitis was established with S. pneumoniae "S" in 65 eyes, S. pneumoniae "I" in 107 eyes, and S. pneumoniae "R" in 78 eyes. Sixteen hours later, control corneas were harvested and the epithelium removed from treatment corneas. Every half hour saline, penicillin, gentamicin, bacitracin, ciprofloxacin, ofloxacin, erythromycin, vancomycin, ceftriaxone, cefotaxime, or chloramphenicol was applied for 5 hours. One hour later CFUs/cornea were assayed. RESULTS: After 24 hours, S. pneumoniae "S" and "I" had proliferated to 9.18+/-6.65 x 10(6) CFUs and 9.26+/-6.90 x 10(6) CFUs. Epithelial removal at 16 hours was not significant. The in vitro antibiotic sensitivity was as expected. However, in vivo, penicillin, gentamicin, or cefazolin sterilized S. pneumoniae "S." S. pneumoniae "R" responded best to fortified gentamicin with or without vancomycin; all others antibiotics were significantly less effective (P < 0.001). CONCLUSIONS: A small intracorneal S. pneumoniae inoculum in rabbit corneas grew and was maintained for 24 hours (with epithelial removal) to provide a model for testing antibiotic sensitivity in vivo. Topical penicillin is best for treating keratitis from penicillin-sensitive S. pneumoniae, whereas topical gentamicin or a combination of gentamicin and vancomycin was most effective against penicillin-resistant S. pneumoniae.
Assuntos
Antibacterianos/uso terapêutico , Infecções Oculares Bacterianas/tratamento farmacológico , Ceratite/tratamento farmacológico , Infecções Pneumocócicas/tratamento farmacológico , Administração Tópica , Animais , Contagem de Colônia Microbiana , Substância Própria/microbiologia , Modelos Animais de Doenças , Quimioterapia Combinada/uso terapêutico , Ceratite/microbiologia , Testes de Sensibilidade Microbiana , Soluções Oftálmicas , Infecções Pneumocócicas/microbiologia , Coelhos , Streptococcus pneumoniae/crescimento & desenvolvimento , Streptococcus pneumoniae/isolamento & purificaçãoRESUMO
BACKGROUND AND OBJECTIVES: Triturated (crushed and suspended) ketoconazole has been recommended for the treatment of fungal keratitis when commercial antifungal eyedrops are unobtainable. The authors evaluated the in vivo corneal stromal concentration with different vehicles in the eyes of adult rabbits. MATERIALS AND METHODS: Ketoconazole and itraconazole tablets were triturated to 20 mg/ml in four vehicles: polyvinyl alcohol (PVA), boric acid, olive oil, and balanced salt solution (BSS). Six eyes (deepithelialized for better penetration) received one drop every 15 minutes for 2 hours. A yeast overlay bioassay of extracts determined the stromal concentration. RESULTS: Itraconazole in BSS, olive oil, PVA, and boric acid produced inhibition zones of 17.3, 15.6, 15.4, and 13.2 mm, respectively. Ketoconazole produced inhibition zones of 35.9, 39.4, 41.8, and 44.7 mm, respectively. From a standard curve, the concentrations of ketoconazole in tissue were 512, 773, 1221, and 1492 micrograms/g, respectively. CONCLUSION: The vehicle that is used to triturate antifungals affects the tissue concentration. This may have an impact on fungal keratitis therapy.