Background: Chemotherapy and radiotherapy can destroy or severely reduce spermatogenesis and thereby jeopardize fertility in the long term. There is still no medical treatment that guarantees fertility preservation after chemo- and radiotherapy. Now with recent improvement of assisted reproductive technologies (ART) and possibility of using testicular spermatozoa or epididymal spermatozoa at in vitro fertilization (IVF) by intracytoplasmic sperm injection (ICSI), cryopreservation of testicular tissue is an option in fertility preservation for males who will lose spermatogenic cells as a result of chemoand radiotherapy and for males with azoospermia. Objectives: This study is an attempt to evaluate oxidative DNA damage in the mice testicular tissue after cryopreservation/thawing cycle when using different types of cryoprotectants as well as to investigate the changes that occur in mice testicular tissue after cryopreservation/thawing cycle as a model for human being. Methods: Fifty mature fertile male mice between 8- 12 weeks old were used in the current study. For each mouse, one testis was evaluated histologically and immunohistochemically (using 8-Hydroxy 2'-Deoxyguanosine as marker of oxidative DNA damage) without cryopreservation (control group). The other testis was evaluated histologically and immunohistochemically (using 8-Hydroxy 2'-Deoxyguanosine as marker of oxidative DNA damage) after six weeks of cryopreservation using different types of cryoprotectants (cryopreserved group). Results: The microscopically observation of slides obtained from cryopreserved testis differs according to the type of cryoprotectant (glycerol, 1,2 propanediol and dimethylsulfoxide), in the dimethylsulfoxide group, tissue sections showed no major differences versus control group, but in the histological sections obtained from tissue cryopreserved by glycerol as cryoprotectant showed moderate morphological and structural changes as compared with the control group. While, the tissue samples subjected to 1,2 propanediol as cryoprotectant displayed the severest morphological and structural changes as compared with the control group. Immunohistochemical results of the present study showed a highly significant (P<0.001) increase in oxidative DNA damage in the cryopreserved testis (46.78%, 63.45% and 32.59% represent cryoprotectant glycerol, 1, 2 propanediol and dimethylsulfoxide, respectively) compared with control group (19.27%) after six weeks of cryopreservation. Conclusion: From the results of the current study, it was concluded that there was alteration in testis histology after cryopreservation/thawing cycle and increase the level of oxidative DNA damage after cryopreservation/thawing cycle. As well as dimethylsulfoxide cryoprotectant provide good protection for testis histology and DNA.
Background: Semen cryopreservation is a useful tool for preserving fertility for men involved in ART program who have been diagnosed with cancer and will undergo chemotherapy, radiotherapy or testicular surgery. Objective: The objective of this study to evaluate the effect of honey bee supplemented to cryoprotectant medium on post-thaw sperm DNA integrity ,morphology and round cells. Subjects ,Materials and Methods: Thirty semen samples were collected from 30 infertile patients. After assessment of semen analysis, semen samples were divided into 3 aliquots (0.7ml for each sample) and mixed with 1 ml of cryopreservation solution (G1,control) alone, enriched with 5% honey bee(G2) with 10% honey bee (G3) for cryopreservation. Current study used the acridine orange test to investigate the effect of honey bee in two concentration (5%, 10%) on the integrity of sperm chromatin structure pre-and post cryopreservation. Cryopreservation was done at -196 Co in liquid nitrogen and thawing was performed after six months. Direct swim up technique was used for in vitro sperm preparation post-thawing, sperm parameters were assessed and data were statistically analyzed pre-and post-thawing. Results: The results show there was a significant(P<0.05) increase in the percentage of morphologically normal sperm for all groups post-thawing, particularly for G3 group that was significantly (P<0.05) increased as compared to G1 and G2 groups, In contrast, non-significant differences (P>0.05) were observed between G1 and G2 groups. Sperm DNA fragmentation percentage (%) was significantly(P<0.05) increased in G1 post thawing without any additives as compared to pre-cryopreservation , where as in G2 and G3 sperm DNA fragmentation were increased but not significantly as post-cryopreservation groups. Also the results show that there were no significant differences between G2 and G3 whereas G3 gives better result as compared to G1 with significant differences. Round cells counts for all groups of post cryopreservation were significantly(P>0.05) decreased as compared to pre-cryopreservation. Non significant (P>0.05) differences in the round cells counts among all the groups of post -thawing in spite of G3 group has the lowest mean. Conclusions: From these results we concluded that supplementation of 10% of honey bee to the freezing cryoprotectant medium gives best protection to the morphology and DNA integrity of human sperm.
Background: The semen cryopreservation has an important role in the male fertility preservation. Certainly, patients who are prone to become infertile due to surgical or medical treatments such as chemo- or radiotherapy for cancer treatment. In fact, semen cryostorage seems to be the only proven method that may offer these couples a chance of having children in the future. Objectives: This study was aimed to investigate the effects of cryopreservation protocols on human sperm parameters. Methods: Sixty semen samples with range of age (19-35) years were included in this study. Sperm parameters, were assessed pre- or post-cryopreservation. Cryopreservation protocols were done using SMART medium with either 15% dimethyl sulfoxide(DMSO) or 15% glycerol alone (as control group) or supplemented with either 0.25M or 0.5M of sucrose for treated groups. Crude data were statistically analyzed. Results: Generally, the results of the present study showed that the sperm parameters were significantly deteriorated post-thawing (P<0.05) as compared to precryopreservation. Both treated groups (G2 and G3) showed best results as compared to G1 (control groups). In contrast, treated group (G2) with 0.25 M of sucrose appeared better results than the as G3 group that treated with 0.5M of sucrose. Conclusion: Sperm parameters including concentration, motility, and morphology were deteriorated after cryopreservation. Using 0.25M sucrose reduced the impact of cryopreservation.
Background: Oocyte vitrification is a promising technique. The choice of appropriate types and concentrations of cryoprotectants is essential for the success of the oocytes vitrification. Objective: This study aimed to investigate the effect of vitrification on viability and morphology of oocytes and to compare the effect of several cryoprotectants on the viability and morphology of oocytes during vitrification and post-thawing. Methods: The sheep ovaries were collected from the local abattoir. Only normal and viable oocytes were included in this study. By using cryotop, immature oocytes that were viable with normal morphology were vitrified with 15% DMSO and 15% EG supplemented with 0.0 M, 0.25M, or 0.5 M of either sucrose or trehalose as control and treated groups. Oocytes viability and morphology were assessed post-aspiration and post-thawing. Results: Fromthe results of the present study, the percentage of post-thawing normal and viable oocyte reported with the use of 0.5M trehalose and EG in vitrification solution (VS) significantly)P<0.05) higher than the percentage of post-thawing normal and viable oocyte reported with use of 0.25 M trehalose and EG. Conclusion: Vitrification is simple technique and easy to perform but it needs some experience to prevent any oocyte loss during vitrification and thawing processing. The use of 0.5 M of either sucrose or trehalose in vitrification solution improves the percentage of post-thawing viable and normal oocytes.