Synthesis, Characterization and Evaluation of Ceftriaxone Mediated BSA Microparticles (CTR–BSA) as a Cryoprotective Agent for Preserving Human Sperm

Infections caused by bacteria may have the potential to regress spermatogenesis, hinder seminal tract and damage sperm. Genital tract infections accounts for 15% of infertility cases among males. Semen extenders supplemented with antimicrobial protein conjugates are used in artificial insemination for the purpose of cryopreserving sperm cells against residual bacterial load and for enhancing the longevity of sperm in in vitro conditions. In this study, an antibiotic mediated protein microparticle was synthesized and analyzed for its antibacterial activity, cytotoxic and genotoxic effects. Ceftriaxone mediated BSA microparticles (CTR–BSA) were prepared by incubating BSA with ceftriaxone (CTR) followed by the intermittent addition of ethanol. Characterization of the microparticle was done using infrared and UV–visible spectroscopy, scanning electron microscope and energy dispersive X-ray and zeta potential analysis. The drug loading efficiency and entrapment efficiency of the microparticle were recorded as 68% and 75% respectively. Continuous dialysis method was performed to check the cumulative percentage of CTR released. Sperm immobilizing capacity of Escherichia coli was confirmed by co-incubating the sperm cells with 24 h grown E. coli culture. In vitro studies of the microparticle at different concentrations were carried out to check its antibacterial activity and cytotoxicity. The microparticle was found to have high antibacterial activity against E. coli and less cytotoxic and genotoxic effects on sperm cells. Molecular docking analysis confirmed minimum binding energy and inhibition constant for the CTR–BSA complex. This study can be incorporated in the development of semen extenders to curb the problem of infectious microbial growth in semen. Graphical Abstract