Effect of Biotin and Folic acid on motility, viability, morphology, chromatin concentration and integrity membrane of sperm in normozoospermia men during Cryopreservation

 Cryopreservation is one of the common techniques in fertility-infertility, which produces reactive oxygen species (ROS) causingsperm cell damage and impairs function and leads to reduced fertility.In diagnosing cryopreserved sperm fertility potential, assessment of various markers has an effective role in the treatment of infertility patients. variable markers of sperm genomic material integrity is costly. In this study, the possible effect of biotin and folic acid on sperm parameters including: motility, viability, morphology, chromatin damage, membrane integrity of frozen and thawed sperm in normozoospermic men has been investigated.

In this experimental study, 30 sperm samples were collected from normozoospermic men aged 25-45 years. [S1] [DD2] Each sample was divided into five parts. Includes fresh pre-cryopreservation group, cryopreservation control groups, biotin (10 mM), folic acid (50 nM) and combination of biotin (10 mM) and folic acid (50 nM). Cryopreserved for two weeks according to the common freezing technique in most infertility reproductive centers at -196 ° C and then thawed. Samples were examined before and after freezing for sperm motility and morphology with computer-aided sperm analysis software. Sperm viability was assessed by eosin-negrosin staining, chromatin density was assessed by toluidine blue (TB) staining and membrane integrity was assessed by HOST (Hypo Osmotic swelling Test) respectively.

Before cryopreservation, motility,viability, morphology, chromatin density quality, sperm membrane integrity washigher in all groups and immotile sperms were lower in all groups than before cryopreservation (p <0.001). After cryopreservation, the quality of chromatin was higher in the folic acid group than in the biotin + folic acid and biotin groups, and all three groups were higher than the control group. The mean sperm life with normal shape in all three groups was higher than the control group. Percentage of sperm with healthy membrane in folic acid group was higher than biotin group and combination of both, and all three groups were more than control(p <0.001). While after cryopreservation the positive correlation was seen between sperm chromatin quality and sperm membrane integrity.

Biotin and folic acid had a protective effect on chromatin quality, healthy membrane, sperm life and played an important role in maintaining sperm parameters after cryopreservation.