Ofloxacin: the next generation of antibiotic in sperm and embryo cultures for assisted reproductive technologies.
OBJECTIVE
To analyze the effect of different concentrations of ofloxacin on sperm kinematic parameters and to determine the embryotoxicity of ofloxacin at physiologic and at 100x concentrations.


METHODS
Prospective comparative study.


METHODS
Clinical and academic research environment.


METHODS
Pooled cryopreserved donor sperm (n = 7).


METHODS
Human sperm were processed through two-layer discontinuous Percoll gradients, and the resultant pellet was resuspended in either HEPES-buffered human tubal fluid medium containing different concentrations of ofloxacin or the control medium. After measuring the kinematic parameters, the percentages of apoptosis and viability were obtained. Next, the sperm DNA was extracted and polymerase chain reaction of beta-globin gene was performed followed by denaturing gradient gel electrophoresis. Mouse embryos recovered at the one-cell pronuclear or zygote stages were cultured in the presence or absence of ofloxacin up to the hatched blastocyst stage and differences in development were recorded.


METHODS
Sperm kinematic parameters, sperm beta-globin gene, and number of embryos reaching the hatched blastocyst stage.


RESULTS
The number of embryos exposed to control and physiologic ofloxacin concentrations showed comparable excellent growth. However, the 100x concentration significantly arrested development. Rates of sperm viability and apoptosis measured 48 hours after exposure to the above concentrations were not different from controls. No differences were noted in the sperm kinematic parameters of sperm exposed to ofloxacin concentrations (1x, 10x, and 100x) or control medium after 0, 4, and 48 hours of incubation. Denaturing gradient gel electrophoresis of beta-globin genes from DNA exposed to varying ofloxacin concentrations failed to show any point mutations.


CONCLUSIONS
Ofloxacin was embryotoxic at pharmacologic concentrations (100x). At physiologic or higher concentrations, ofloxacin appears to be safe and does not affect sperm kinematic parameters when compared with controls. This may indicate that sperm motility parameters alone cannot be relied on to evaluate the effects of drugs on fertility and that in vitro embryologic studies are essential. Ofloxacin at any concentration did not alter the rates of sperm apoptosis or viability. Ofloxacin does not appear to be mutagenic as evidenced by the beta-globin gene analysis.
Erhart
B
B
Department of Gynecology and Obstetrics, Loma Linda University School of Medicine, California 92350, USA.
Chan
P J
PJ
Patton
W C
WC
King
A
A
D000704:Analysis of Variance; D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D015925:Cryopreservation; D004247:DNA / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005314:Embryonic and Fetal Development / Q000187:drug effects; D005260:Female; D005914:Globins / Q000032:analysis / Q000235:genetics; D006801:Humans; D008297:Male; D051379:Mice; D009154:Mutation; D015242:Ofloxacin / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D011446:Prospective Studies; D013081:Sperm Motility / Q000187:drug effects* / Q000502:physiology; D013084:Sperm-Ovum Interactions; D013094:Spermatozoa / Q000737:chemistry / Q000187:drug effects* / Q000502:physiology; D013997:Time Factors
D000890:Anti-Infective Agents; D005914:Globins; D004247:DNA; D015242:Ofloxacin