Evaluation of sperm’s chromatin quality with acridine orange test, chromomycin A3 and aniline blue staining in couples with unexplained recurrent abortion

Abstract

Objective To evaluate the sperm's chromatin quality in couples with spontaneous recurrent abortion. Methods Thirty couples with spontaneous recurrent abortion (case group) and 30 fertile couples (control group) referring to Zeinabieh Gynecology clinic of Shiraz were included. Semen samples were collected for each participant and were used for standard semen analysis and sperm nuclear maturity tests including Chromomycin A3 (CMA3), Aniline Blue (AB) staining and Acridine Orange (AO) test (by light microscopy).Result Patients in case group had significantly higher percentage of CMA3 (p < 0.001) and AB (p <0.001) positive spermatozoa compared to controls. However AO results did not differ significantly between groups (p= 0.656). Sperm morphology and progressive motility were negatively correlated with CMA3 (p=0.001 and p=0.043) and AB (p=0.015 and p=0.031) respectively. Conclusion Evaluation of the sperm's quality via CMA3 and AB staining could be considered as one of the complementary tests of semen analysis for assessment of male factor in couples with spontaneous recurrent abortion.

Use of Silver Nitrate for the Assessment of Sperm Measurements in Selected Farm and Free-Living Animal Species

“…The most frequently assessed, and the most important semen parameters include concentration, motility, and morphology of the sperm (13). These parameters are considered to be the most useful since they have been correlated with the fertility potential (14,17,18). For example, low sperm concentration, which is connected with the number of spermatozoa present in one millimetre of semen, indicates male infertility (11).…”

“…This is because standard staining techniques applied for the identification of semen quality, and assessment of spermatozoa are in the majority of cases limited to detection of morphological abnormality only (17,18,24), while sperm chromatin structure is of a crucial importance for fertilisation and embryo development. Routine semen assessments are the first step in the evaluation of male infertility factors, but this analysis is not considered to be sufficient for determining infertility in in vivo and in vitro conditions (17). Consequently, the ultimate determination of infertility often cannot be performed on the basis of the routine semen assessment (24).…”

“…Consequently, the ultimate determination of infertility often cannot be performed on the basis of the routine semen assessment (24). The lack of conventional sperm parameters that would enable to predict fertilisation efficacy suggests that there might be latent abnormalities precisely at the level of sperm chromatin (17). Over the last decade there have been significant advances made in studies on the role of nuclear sperm DNA integrity in the analysis of male infertility factors.…”

“…An expanding area of interest in the field of sperm chromatin condensation involves explaining whether, and in what way protamination disturbances affect the fertilisation capacity of a spermatozoon (5,16). Hence, the application of indirect methods of assessing protamin numbers and chromatin structure (DNA or chromatin integrity) measurements on the basis of various staining procedures has become increasingly more common (15,17).…”

Abnormal Chromatin Condensation in Spermatozoa and Dna Fragmentation in Spermatozoa: Is There a Correlation?

“…Эти данные позволяют сделать заключение, что нарушение конденсации хроматина может служить предиктивным показателем аномалий развития эмбри-она как in vivo, так и in vitro. Причем негативные по-следствия нарушения компактизации проявляются в раннее время после оплодотворения [77], приводя к раннему эмбриональному аресту [78] и спонтанным абортам [27,79].…”