Acridine Orange and Flow Cytometry: Which Is Better to Measure the Effect of Varicocele on Sperm DNA Integrity?

Abstract

We evaluated the effect of varicocelectomy on semen parameters and levels of sperm DNA damage in infertile men. A total of 75 infertile men with varicocele and 40 fertile men (controls) were included in this study. Semen analysis and sperm DNA damage expressed as the DNA fragmentation index using acridine orange staining and chromatin condensation test by flow cytometry were assessed before and 6 months after varicocelectomy. The patients were also followed up for 1 year for pregnancy outcome. Semen parameters were significantly lower in varicocele patients compared to controls (P < 0.05). Mean percentages of sperm DNA fragmentation and sperm DNA chromatin condensation in patients were significantly higher than those in controls (P < 0.05). After varicocelectomy, sperm DNA fragmentation improved significantly, whereas sperm chromatin condensation was not significantly changed. In 15 out of 75 varicocele patients, clinical pregnancy was diagnosed; those with positive pregnancy outcome had significant improvement in sperm count, progressive sperm motility, and sperm DNA fragmentation, but there was no significant difference in sperm DNA condensation compared to negative pregnancy outcome patients. We concluded from this study that acridine orange stain is more reliable method than flow cytometry in the evaluation of sperm DNA integrity after varicocelectomy.

Impact of leukocytospermia on sperm dynamic motility parameters, DNA and chromosomal integrity

“…Despite some debate regarding its accuracy, acridine orange test (AOT) is a simple and rapid test that is still applied in the evaluation of sperm DNA integrity of infertile men with different etiologies. Its results are comparable to the other more advanced tests such as TUNEL and flowcytometry [10, 11].…”

Effect of Venlafaxine, Pramipexole, and Valsartan on Spermatogenesis in Male Rats

“…The DNA fragmentation index (DFI) was calculated by using the following formula: A DFI value <30% was considered as normal. 36 …”

Sperm DNA Fragmentation: A New Guideline for Clinicians

“…The findings of higher SDF in both fertile and infertile men with varicocele than controls [ 168 ] and significant decrease in SDF levels after varicocele repair [ 261 ] provide the rationale of SDF testing in refining the selection of varicocelectomy candidates. In addition, reduction in SDF seems to translate into better reproductive outcomes [ 262 263 264 ]. Although the association between SDF and high-grade varicocele is much stronger, patients with low-grade varicocele had achieved improvement in natural pregnancy rate that were similar to those with high-grade varicocele after surgery [ 265 ].…”

“…On the contrary, studies have shown that a number of interventions can be utilized in this regard. Examples of such interventions include recurrent ejaculation to shorten the abstinence time [ 222 227 350 351 352 ], oral antioxidant therapy [ 205 207 208 353 354 355 356 357 358 359 360 361 362 ], performing varicocelectomy for patients with clinical varicocele [ 168 176 177 179 263 264 360 363 364 365 366 367 368 369 370 371 372 373 374 375 376 377 378 379 380 381 382 ], treating genitourinary infections when diagnosed [ 47 ], and utilizing advanced sperm selection techniques for ICSI such as PICSI/IMSI [ 383 384 385 ] or using testicular sperm instead of ejaculated sperm [ 251 252 269 386 387 ].…”