Andrology: The use of hemizona assay in the evaluation of the optimal sperm preparation technique
TL;DR: The results support the conclusion that in-vitro sperm preparation methods can affect sperm binding to the zona pellucida and the hemizona assay can be used in selecting the optimal sperm preparation method prior to its use for assisted reproductive techniques.
Abstract: The objective of the study was to evaluate the benefit of different sperm preparation methods by using the hemizona assay. A total of 58 men admitted to the male infertility clinic for evaluation were tested by routine semen analysis and hemizona assay. Five different techniques (swim-up, TEST-yolk buffer, Percoll, pentoxifylline and progesterone) were used for preparation of sperm suspensions. The effect of these treatments on the sperm-binding capacity using the hemizona assay was assessed. The routine swim-up preparation was used as the reference method. Of the four preparation methods, only the TEST-yolk buffer and pentoxifylline exhibited an overall statistically significant improvement in sperm-binding capacity in comparison with the swim-up preparation method (P = 0.01 and 0.001 respectively). Following preparation with Percoll and progesterone there was no change in the mean value of binding capacity, compared with swim-up. However, examination of the effect of the four treatments on each specimen individually yielded a diversity in the response, e.g. having the capability to enhance, damage or be ineffective in sperm binding capacity. The results support the conclusion that in-vitro sperm preparation methods can affect sperm binding to the zona pellucida. Since there is a diversity in the response of sperm samples to different treatments, the hemizona assay can be used in selecting the optimal sperm preparation method prior to its use for assisted reproductive techniques. This is advocated mainly for the 'male factor' group.
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TL;DR: There is insufficient evidence to recommend any specific preparation technique, and large high quality randomised controlled trials, comparing the effectiveness of a gradient and/or a swim-up and/ or wash and centrifugation technique on clinical outcome are lacking.
Abstract: Background Semen preparation techniques for assisted reproduction, including intrauterine insemination (IUI), were developed to separate the motile morphological normal spermatozoa. Leucocytes, bacteria and dead spermatozoa produce oxygen radicals that negatively influence the ability to fertilize the egg. The yield of as many motile, morphologically normal spermatozoa as possible might influence treatment choices and therefore outcomes. Objectives To compare the effectiveness of gradient, swim-up, or wash and centrifugation semen preparation techniques on clinical outcome in subfertile couples undergoing intrauterine insemination (IUI). Search strategy We searched the Menstrual Disorders and Subfertility Group Trials Register (3 January 2007), Cochrane Central Register of Controlled Trials (CENTRAL) (The Cochrane Library 2007, Issue 2), MEDLINE (1966 to January 2007), EMBASE (1980 to January 2007), Science Direct Database (1966 to January 2007), National Research Register (2000 to 2007), Biological Abstracts (2000 to January 2007), CINAHL (1982 to October 2006) and reference lists of relevant articles. We also contacted experts and authors in the field. Selection criteria Parallel randomized controlled trials (RCTs) comparing the efficacy of semen preparation techniques used for subfertile couples undergoing IUI in terms of clinical outcome were included. Data collection and analysis Two reviewer authors independently assessed trial quality and extracted data. Study authors were contacted for additional information. Main results Five RCTs, including 262 couples in total, were included in the meta-analysis (Dodson 1998; Grigoriou 2005; Posada 2005; Soliman 2005; Xu 2000). Xu compared the three techniques; Soliman compared a gradient technique versus a wash technique; Dodson and Posada compared a gradient technique versus a swim-up technique; whereas Grigoriou compared swim-up versus a wash technique. No trials reported the primary outcome of live birth. There was no evidence of a difference between pregnancy rates (PR) for swim-up versus a gradient or wash and centrifugation technique (Peto OR 1.57, 95% CI 0.74 to 3.32; Peto OR 0.41, 95% CI 0.15 to 1.10, respectively); nor in the two studies comparing a gradient technique versus wash and centrifugation (Peto OR 1.76, 95% CI 0.57 to 5.44). There was no evidence of a difference in them is carriage rate (MR) in two studies comparing swim- up versus a gradient technique (Peto OR 0.13, 95% CI 0.01 to 1.33). Authors' conclusions There is insufficient evidence to recommend any specific preparation technique. Large high quality randomised controlled trials, comparing the effectiveness of a gradient and/or a swim-up and/or wash and centrifugation technique on clinical outcome are lacking. Further randomised trials are warranted.
179 citations
TL;DR: The carbohydrate moieties derived from ZIF-1 reduced the binding of native Zif-1 on human spermatozoa as well as the zona binding inhibitory activity of the glycoprotein, although the intensity of the effects are lower when compared with the native protein.
46 citations
TL;DR: It is demonstrated that black-footed ferret semen has a comparatively high osmolality and sperm motility was more susceptible to hyperosmotic conditions than acrosomal integrity, and neither were influenced by hypotonic solutions.
37 citations
TL;DR: The objective of this study was to validate the HZA for canine spermatozoa, as a test for diagnosing canine male fertility status, and to avoid the effect of bad zona on sperm binding.
30 citations
TL;DR: The present study reports three factors that affect HZA: the base medium used, the protein source and the size of pipette used for removing loosely bound spermatozoa during HZA, which emphasize the importance of standardization of the protocol of the hemizona assay worldwide to be able to compare results between different laboratories.
Abstract: 'To whom correspondence should be addressed Sperm binding to the zona pellucida is a prerequisite for fertilization. The hemizona binding assay (HZA) is commonly used to evaluate the zona-binding capacity of spermatozoa. The present study reports three factors that affect HZA. They were the base medium used, the protein source and the size of pipette used for removing loosely bound spermatozoa during HZA. The number of spermatozoa bound on the hemizona was compared between (1) Earle's balanced salt solution (EBSS) and Ham's F-10, and (2) between human serum and bovine serum albumin (BSA). Results indicated that EBSS and human serum both significantly increase the number of bound spermatozoa when compared to Ham's F-10 (P < 0.0001, paired Mest) and BSA (P < 0.05, paired Mest) respectively. Pipettes of different diameters were used to study the effect of size in removing loosely bound spermatozoa on hemizona. Data showed that the diameter of the pipette should be 2*200 mm, in order not to remove bound spermatozoa excessively. These results emphasize the importance of standardizati on of the protocol of the hemizona assay worldwide to be able to compare results between different laboratories.
25 citations
References
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TL;DR: This study demonstrates no therapeutic advantage of using pentoxifylline in IVF for male factor infertility in cases with previous fertilization failure and no significant differences between treatment and control groups.
37 citations
TL;DR: It is concluded that cryopreservation results in a reduced number of motile sperm cells but does not adversely affect the ability of rescued sperm cells to bind to the ZP.
27 citations
"Andrology: The use of hemizona assa..." refers methods in this paper
...A cryopreserved pool of six ejaculates from different donors served as a control sample for all the hemizona assays performed in this study (Gamzu et al., 1992)....
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...Leitz micromanipulators (Leica, Wetzlar, Germany) were used for cutting the oocytes as described previously (Burkman et al, 1988; Gamzu et al., 1992)....
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