Sperm DNA fragmentation (SDF) has been generally acknowledged as a valuable tool for male fertility evaluation. While its detrimental implications on sperm function were extensively investigated, little is known about the actual indications for performing SDF analysis. This review delivers practice based recommendations on commonly encountered scenarios in the clinic. An illustrative description of the different SDF measurement techniques is presented. SDF testing is recommended in patients with clinical varicocele and borderline to normal semen parameters as it can better select varicocelectomy candidates. High SDF is also linked with recurrent spontaneous abortion (RSA) and can influence outcomes of different assisted reproductive techniques. Several studies have shown some benefit in using testicular sperm rather than ejaculated sperm in men with high SDF, oligozoospermia or recurrent in vitro fertilization (IVF) failure. Infertile men with evidence of exposure to pollutants can benefit from sperm DNA testing as it can help reinforce the importance of lifestyle modification (e.g., cessation of cigarette smoking, antioxidant therapy), predict fertility and monitor the patient's response to intervention.

OBJECTIVE

To compare, in a retrospective study, the ultrasound findings and hormonal changes after testicular sperm extraction (TESE) using the conventional multiple biopsy approach and the more recent microdissection technique. TESE has been performed using the conventional multiple biopsy approach and the more recent microdissection technique.

METHODS

A total of 435 men with nonobstructive azoospermia who had undergone 543 TESE attempts were included in the study. The initial 83 attempts were done using the conventional open technique and the remaining 460 attempts were performed by microdissection. The sperm retrieval rates were compared, as were the complication rates as assessed by ultrasound and endocrinologic evaluations between the two groups.

RESULTS

The retrieval rate by the conventional technique was 32% and by microdissection was 57% (P = 0.0002). In patients with hypospermatogenesis, the retrieval rate differed between the two approaches (P = 0.03). Ultrasound findings demonstrated fewer acute and chronic changes in the microdissection group than in the conventional group (P < 0.05). At 3 to 6 months after surgery, the testosterone levels had dropped to 80% of their pre-TESE levels in both groups (P < 0.01). The levels rose back to 85% after 12 months and to 95% after 18 months. The mean follicle-stimulating hormone levels increased from 22 +/- 2 to 30 +/- 3 IU/L (P = 0.02), and the luteinizing hormone levels increased from 12 +/- 2 to 16 +/- 2 IU/L (P = 0.2).

CONCLUSIONS

TESE has effects on testicular function, but the microdissection procedure is relatively safer than the conventional technique and improves the sperm retrieval rate significantly in patients with nonobstructive azoospermia.

OBJECTIVE

We examined preoperative factors that could predict successful microdissection testicular sperm extraction in men with azoospermia and nonmosaic Klinefelter's syndrome. We also analyzed the influence of preoperative hormonal therapy on the sperm retrieval rate.

METHODS

A total of 91 microdissection testicular sperm extraction attempts were done in 68 men with nonmosaic Klinefelter's syndrome. Men with serum testosterone less than 300 ng/dl received medical therapy with aromatase inhibitors, clomiphene or human chorionic gonadotropin before microdissection testicular sperm extraction. Preoperative factors of patient age and endocrinological data were compared in those in whom the procedure was and was not successful. The sperm retrieval rate was the main outcome. Clinical pregnancy (pregnancy with heartbeat) and the live birth rate were also calculated.

RESULTS

Testicular spermatozoa were successfully retrieved in 45 men (66%), representing 62 (68%) attempts. Increasing male age was associated with a trend toward a lower sperm retrieval rate (p = 0.05). The various types of preoperative hormonal therapies did not have different sperm retrieval rates but men with normal baseline testosterone had the best sperm retrieval rate of 86%. Patients who required medical therapy and responded to that treatment with a resultant testosterone of 250 ng/dl or higher had a higher sperm retrieval rate than men in whom posttreatment testosterone was less than 250 ng/dl (77% vs 55%). For in vitro fertilization attempts in which sperm were retrieved the clinical pregnancy and live birth rates were 57% and 45%, respectively.

CONCLUSIONS

Microdissection testicular sperm extraction is an effective sperm retrieval technique in men with Klinefelter's syndrome. Men with hypogonadism who respond to medical therapy may have a better chance of sperm retrieval.

OBJECTIVE

To evaluate the outcomes of microdissection testicular sperm extraction (micro-TESE) in patients with high FSH.

METHODS

Clinical retrospective study.

METHODS

Department of urology at a tertiary university hospital.

METHODS

Seven hundred ninety-two men with nonobstructive azoospermia.

METHODS

Micro-TESE followed by intracytoplasmic sperm injection was performed. The men were classified into four groups based on serum FSH levels: <15, 15-30, 31-45, and >45 IU/mL.

METHODS

Sperm retrieval, clinical pregnancy, and live birth rates.

RESULTS

Testicular sperm were successfully retrieved in 60% of the men. Sperm retrieval rates in the groups of men with FSH values 15-30, 31-45, and >45 IU/mL was 60%, 67%, and 60% respectively; this was higher than the group of men with FSH < 15 (51%). Of those men who had sperm retrieved, clinical pregnancy and live birth rates were similar in the four groups (46%, 50%, 52%, 46% and 38%, 45%, 44%, 36%, respectively).

CONCLUSIONS

The chances of sperm retrieval using micro-TESE is just as common, if not better for men with elevated FSH levels than for men with lower FSH. Micro-TESE results appear to differ from earlier series that report low retrieval rates with random biopsies for men with elevated FSH. High FSH is not a contraindication for micro-TESE.

Traditionally, testosterone and estrogen have been considered to be male and female sex hormones, respectively. However, estradiol, the predominant form of estrogen, also plays a critical role in male sexual function. Estradiol in men is essential for modulating libido, erectile function, and spermatogenesis. Estrogen receptors, as well as aromatase, the enzyme that converts testosterone to estrogen, are abundant in brain, penis, and testis, organs important for sexual function. In the brain, estradiol synthesis is increased in areas related to sexual arousal. In addition, in the penis, estrogen receptors are found throughout the corpus cavernosum with high concentration around neurovascular bundles. Low testosterone and elevated estrogen increase the incidence of erectile dysfunction independently of one another. In the testes, spermatogenesis is modulated at every level by estrogen, starting with the hypothalamus-pituitary-gonadal axis, followed by the Leydig, Sertoli, and germ cells, and finishing with the ductal epithelium, epididymis, and mature sperm. Regulation of testicular cells by estradiol shows both an inhibitory and a stimulatory influence, indicating an intricate symphony of dose-dependent and temporally sensitive modulation. Our goal in this review is to elucidate the overall contribution of estradiol to male sexual function by looking at the hormone's effects on erectile function, spermatogenesis, and libido.

Traditionally, the success of a researcher is assessed by the number of publications he or she publishes in peer-reviewed, indexed, high impact journals. This essential yardstick, often referred to as the impact of a specific researcher, is assessed through the use of various metrics. While researchers may be acquainted with such matrices, many do not know how to use them to enhance their careers. In addition to these metrics, a number of other factors should be taken into consideration to objectively evaluate a scientist's profile as a researcher and academician. Moreover, each metric has its own limitations that need to be considered when selecting an appropriate metric for evaluation. This paper provides a broad overview of the wide array of metrics currently in use in academia and research. Popular metrics are discussed and defined, including traditional metrics and article-level metrics, some of which are applied to researchers for a greater understanding of a particular concept, including varicocele that is the thematic area of this Special Issue of Asian Journal of Andrology. We recommend the combined use of quantitative and qualitative evaluation using judiciously selected metrics for a more objective assessment of scholarly output and research impact.

OBJECTIVE

Although optimizing endogenous testosterone production before testicular sperm extraction is commonly practiced, whether improved preoperative testosterone levels enhance sperm retrieval remains unclear. We evaluated the influence of preoperative medical therapy in men with nonobstructive azoospermia before microdissection testicular sperm extraction.

METHODS

A total of 1,054 men underwent microdissection testicular sperm extraction from 1999 to 2010. Patients with preoperative testosterone levels less than 300 ng/dl were treated with aromatase inhibitors, clomiphene citrate or human chorionic gonadotropin before microdissection testicular sperm extraction with the goal of optimizing testosterone levels. Patient demographics, preoperative testosterone levels, sperm retrieval rate and pregnancy outcomes were recorded and compared in men with different baseline testosterone levels.

RESULTS

Of the 736 men who had preoperative hormonal data 388 (53%) had baseline testosterone levels greater than 300 ng/dl. The sperm retrieval rate in these men was 56%. In the remaining 348 men with pretreatment testosterone levels less than 300 ng/dl, the sperm retrieval rate was similar (52%, p = 0.29). In addition, the sperm retrieval, clinical pregnancy and live birth rates were similar between men who responded to hormonal therapy and those who did not.

CONCLUSIONS

Men with nonobstructive azoospermia and hypogonadism often respond to hormonal therapy with an increase in testosterone levels, but neither baseline testosterone level nor response to hormonal therapy appears to affect overall sperm retrieval, clinical pregnancy or live birth rates.

OBJECTIVE

To investigate the relative differences in outcomes among microdissection testicular sperm extraction (micro-TESE), conventional testicular sperm extraction (cTESE), and testicular sperm aspiration (TESA) in men with nonobstructive azoospermia.

METHODS

Systematic review and meta-analysis.

METHODS

Outpatient academic and private urology clinics.

METHODS

Men with nonobstructive azoospermia.

METHODS

Micro-TESE, cTESE, or TESA.

METHODS

Sperm retrieval (SR).

RESULTS

Fifteen studies with a total of 1,890 patients were identified. The weighted average age of the patients was 34.4 years, the follicular stimulating hormone level was 20.5 mIU/mL, the T was 373 ng/dL, and the testicular volume was 13.5 mL. In a direct comparison, performance of micro-TESE was 1.5 times more likely (95% confidence interval 1.4-1.6) to result in successful SR as compared with cTESE. Similarly, in a direct comparison, performance of cTESE was 2.0 times more likely (95% confidence interval 1.8-2.2) to result in successful SR as compared with TESA. Because of inconsistent reporting, evaluation of other procedural characteristics and pregnancy outcomes was not possible.

CONCLUSIONS

Sperm retrieval was higher for micro-TESE compared with cTESE and for cTESE compared with TESA. Standardization of reported outcomes as well as combining all available SR data would help to further elucidate the SRs of these procedures.

Despite advances in the field of male reproductive health, idiopathic male infertility, in which a man has altered semen characteristics without an identifiable cause and there is no female factor infertility, remains a challenging condition to diagnose and manage. Increasing evidence suggests that oxidative stress (OS) plays an independent role in the etiology of male infertility, with 30% to 80% of infertile men having elevated seminal reactive oxygen species levels. OS can negatively affect fertility via a number of pathways, including interference with capacitation and possible damage to sperm membrane and DNA, which may impair the sperm's potential to fertilize an egg and develop into a healthy embryo. Adequate evaluation of male reproductive potential should therefore include an assessment of sperm OS. We propose the term Male Oxidative Stress Infertility, or MOSI, as a novel descriptor for infertile men with abnormal semen characteristics and OS, including many patients who were previously classified as having idiopathic male infertility. Oxidation-reduction potential (ORP) can be a useful clinical biomarker for the classification of MOSI, as it takes into account the levels of both oxidants and reductants (antioxidants). Current treatment protocols for OS, including the use of antioxidants, are not evidence-based and have the potential for complications and increased healthcare-related expenditures. Utilizing an easy, reproducible, and cost-effective test to measure ORP may provide a more targeted, reliable approach for administering antioxidant therapy while minimizing the risk of antioxidant overdose. With the increasing awareness and understanding of MOSI as a distinct male infertility diagnosis, future research endeavors can facilitate the development of evidence-based treatments that target its underlying cause.

We present a case of familial prune belly syndrome and review potential modes of inheritance for the syndrome. A total of 11 cases of familial prune belly syndrome have been reported. These cases were reviewed and possible modes of inheritance were determined for each case. Our review strongly suggests a sex-influenced autosomal recessive mode of inheritance.

OBJECTIVE

We compared satisfaction and treatment efficacy in men with symptomatic hypogonadism who received clomiphene citrate or testosterone supplementation therapy.

METHODS

Men treated with clomiphene citrate, or testosterone injections or gels for symptomatic hypogonadism (total testosterone less than 300 ng/dl) reported satisfaction with the current treatment regimen using the qADAM questionnaire.

RESULTS

A total of 93 men on testosterone injections (31) or gels (31), or clomiphene citrate (31) were age matched from a retrospective cohort of 1,150 on testosterone supplementation therapy. We compared men who received testosterone supplementation therapy to 31 not on such therapy, who served as controls. Median serum testosterone increased from pretreatment levels in all men regardless of therapy with clomiphene citrate, and testosterone injections and gels (from 247 to 504, 224 to 1,104 and 230 to 412 ng/dl, respectively, p <0.05). Final median serum total testosterone in men on clomiphene citrate (504 ng/dl) was lower than in men receiving testosterone injections (1,014 ng/dl, p <0.01) but similar to that in men on testosterone gels (412 ng/dl, p = 0.31). Despite different serum testosterone levels men on all 3 therapies reported similar satisfaction on qADAM, including a score of 35 for clomiphene citrate, 39 for testosterone injections, 36 for testosterone gels and 34 for control treatment (p >0.05). Men receiving testosterone injections reported greater libido than men on clomiphene citrate (4 vs 3, p = 0.04) or testosterone gels (4 vs 3, p = 0.04), or controls (4 vs 3, p <0.01).

CONCLUSIONS

Testosterone supplementation regimens and clomiphene citrate are efficacious for improving serum total testosterone. No difference in overall hypogonadal symptoms was noted among men on any testosterone supplementation therapy. Despite lower serum total testosterone, men on clomiphene citrate and testosterone gels reported satisfaction similar to that of men treated with testosterone injections.

OBJECTIVE

We developed an artificial neural network and nomogram using readily available clinical features to model the chance of identifying sperm with microdissection testicular sperm extraction by readily available preoperative clinical parameters for men with nonobstructive azoospermia.

METHODS

We reviewed the records of 1,026 men who underwent microdissection testicular sperm extraction. Patient age, follicle-stimulating hormone level, testicular volume, history of cryptorchidism, Klinefelter syndrome and presence of varicocele were included in the models. For the artificial neural network the data set was divided randomly into a training set (75%) and a test set (25%) with n1/n2 cross validation used to evaluate model accuracy, and then modeled with a neural computational system. In addition, a nomogram with calibration plots was developed to predict sperm retrieval with microdissection testicular sperm extraction. We compared these models to logistic regression.

RESULTS

The ROC area for the neural computational system in the test set was 0.641. The neural network correctly predicted the outcome in 152 of the 256 test set patients (59.4%). The nomogram AUC was 0.59 and adequately calibrated. Multivariable logistic regression demonstrated patient age, history of Klinefelter syndrome and cryptorchidism to be significant predictors of sperm retrieval (p <0.05). However, follicle-stimulating hormone and testicular volume were not significant by internal validation.

CONCLUSIONS

We modeled a combination of well described preoperative clinical parameters to predict sperm retrieval using a neural computational system and nomogram with acceptable predictive values. The generalizability of these findings requires external validation.

Varicocele is a pathologic enlargement of the pampiniform venous plexus within the spermatic cord, a condition that is a common cause of impaired sperm production and decreased quality of sperm. While varicocele is the most common surgically correctable risk factor for male infertility, not all males with varicocele experience infertility. In fact, most men with varicocele have normal spermatogenesis. Despite its prevalence, the molecular mechanisms of varicocele and its effect on testicular function are yet to be completely understood. We postulate that men with varicocele-associated infertility could have preexisting genetic lesions or defects in molecular mechanisms that make them more susceptible to varicocele-mediated testicular injury affecting spermatogenesis.

OBJECTIVE

Men with azoospermia and severe testicular atrophy may be counseled to avoid sperm retrieval due to perceived limited success. We evaluated the outcomes of microdissection testicular sperm extraction in men with severe testicular atrophy (volume 2 ml or less).

METHODS

We reviewed the records of 1,127 men with nonobstructive azoospermia who underwent microdissection testicular sperm extraction followed by intracytoplasmic sperm injection. They were classified into 3 groups based on average testicular volume, including 2 ml or less, greater than 2 to less than 10 and 10 or greater. Sperm retrieval, clinical pregnancy and live birth rates were calculated. Clinical features evaluated included age, follicle-stimulating hormone level, cryptorchidism history, Klinefelter syndrome, varicocele and testicular histology on diagnostic biopsy.

RESULTS

Testicular sperm were successfully retrieved in 56% of the men. The sperm retrieval rate in those with a testicular volume of 2 ml or less, greater than 2 to less than 10 and 10 or greater was 55%, 56% and 55%, respectively. Clinical pregnancy and live birth rates were similar in men in the 3 groups who underwent sperm retrieval (55.2%, 50.0% and 47.0%, and 47.2%, 43.0% and 42.2%, respectively). Of the 106 men with an average testis volume of 2 ml or less those from whom sperm were retrieved were younger (31.1 vs 35.2 years) and more likely to have a history of Klinefelter syndrome (82.2% vs 55.6%) than men in whom sperm were not found (p <0.05). Men in this group had a higher prevalence of Klinefelter syndrome than men with a testis volume of greater than 2 ml (72.6% vs 5.3%, p <0.0001). Men younger than 30 years with Klinefelter syndrome had a higher sperm retrieval rate than men older than 30 years without Klinefelter syndrome (81.8% vs 33%, p <0.01). There was no cutoff point for age beyond which sperm could not be retrieved in men with small testes. On multivariable analysis younger age was the only preoperative factor associated with successful sperm retrieval in men with small testes (2 ml or less).

CONCLUSIONS

Testicular volume does not affect the sperm retrieval rate at our center for microdissection testicular sperm extraction. Of men with the smallest volume testes those who were younger with Klinefelter syndrome had the highest sperm retrieval rate. Severe testicular atrophy should not be a contraindication to microdissection testicular sperm extraction.

OBJECTIVE

To report the chance of sperm discovery in the laboratory when sperm were not identified in the operating room (OR).

METHODS

Clinical retrospective study.

METHODS

Department of urology at a tertiary university hospital.

METHODS

A total of 1,054 men with nonobstructive azoospermia who underwent microdissection testicular sperm extraction.

METHODS

Preoperative and intraoperative parameters were analyzed relative to the chance of sperm identification using a tissue digestion protocol in the laboratory if no sperm were observed in the OR.

METHODS

Sperm retrieval, clinical pregnancy, and live birth rates.

RESULTS

Sperm were found in the OR in 52.5% of the 1,054 men. Of the 501 men for whom sperm were not identified by andrologists in the OR, sperm were found in the laboratory for an additional 35 (7%). On multivariable logistic regression analysis, the presence of germ cells intraoperatively was the only predictor of identifying sperm in the laboratory after tissue digestion.

CONCLUSIONS

In men undergoing microdissection testicular sperm extraction, when sperm were not observed in the OR despite extensive mechanical processing, sperm were observed in the laboratory for 7% of the men. This information is valuable in counseling couples in the immediate postoperative period when no sperm were identified intraoperatively.

Low-intensity extracorporeal shock wave therapy (Li-ESWT) has been proposed as an effective non-invasive treatment option for erectile dysfunction (ED).

To use systematic review and meta-analysis to assess the efficacy of Li-ESWT by comparing change in erectile function as assessed by the erectile function domain of the International Index of Erectile Function (IIEF-EF) in men undergoing Li-ESWT vs sham therapy for the treatment of ED.

Systematic search was conducted of MEDLINE, EMBASE, and ClinicalTrials.gov for randomized controlled trials that were published in peer-reviewed journals or presented in abstract form of Li-ESWT used for the treatment of ED from January 2010 through March 2016. Randomized controlled trials were eligible for inclusion if they were published in the peer-reviewed literature and assessed erectile function outcomes using the IIEF-EF score. Estimates were pooled using random-effects meta-analysis.

Change in IIEF-EF score after treatment with Li-ESWT in patients treated with active treatment vs sham Li-ESWT probes.

Data were extracted from seven trials involving 602 participants. The average age was 60.7 years and the average follow-up was 19.8 weeks. There was a statistically significant improvement in pooled change in IIEF-EF score from baseline to follow-up in men undergoing Li-ESWT vs those undergoing sham therapy (6.40 points; 95% CI = 1.78-11.02; I2 = 98.7%; P < .0001 vs 1.65 points; 95% CI = 0.92-2.39; I2 = 64.6%; P < .0001; between-group difference, P = .047). Significant between-group differences were found for total treatment shocks received by patients (P < .0001).

In this meta-analysis of seven randomized controlled trials, treatment of ED with Li-ESWT resulted in a significant increase in IIEF-EF scores.

OBJECTIVE

We determined the effect of prior biopsies with no sperm seen on the chance of sperm retrieval with microdissection testicular sperm extraction in men with nonobstructive azoospermia.

METHODS

A total of 311 men with NOA underwent microdissection testicular sperm extraction. Of these patients 135 underwent no prior biopsies, 159 underwent 1 or 2 diagnostic testicular biopsies per testis and 17 underwent 3 or 4. The outcome measure studied was the success of sperm retrieval with microdissection testicular sperm extraction. Serum follicle-stimulating hormone and histopathological diagnosis were examined as predictive factors for sperm recovery.

RESULTS

Spermatozoa were retrieved in 150 men by microdissection testicular sperm extraction (48%). The success of sperm retrieval in patients who underwent 3 to 4 biopsies (23%) was lower than the retrieval rate in patients who underwent no prior biopsies (56%) and 1 to 2 biopsies per testis (51%) (p = 0.04). When histopathology was considered, patients with Sertoli-cell-only diagnosis on prior diagnostic biopsy had lower retrieval rates compared to the group with no biopsies (p = 0.02). Men with maturation arrest and hypospermatogenesis had similar microdissection testicular sperm extraction sperm retrieval rates regardless of the number of prior biopsies. Spermatozoa recovery was independent of serum follicle-stimulating hormone.

CONCLUSIONS

There is no threshold of prior negative biopsies that precludes the success of sperm retrieval using microdissection testicular sperm extraction. A limited number of testicular biopsies provide limited or no prognostic value for sperm retrieval with microdissection testicular sperm extraction.

OBJECTIVE

To evaluate the effect of the delivery of testis on semen parameters, serum testosterone, and pregnancy rates after varicocelectomy, performed with and without delivery of the testis.

METHODS

A total of 165 patients with clinically diagnosed varicoceles who underwent surgical ligation with (n = 55) or without (n = 110) testicular delivery were included in this study. Patients were followed up postoperatively at 3, 6, and 9 to 24 months. The postoperative outcome was assessed by determination of the semen parameters, testosterone level, varicocele recurrence, and pregnancy rate.

RESULTS

No varicocele recurrences were clinically detectable after either surgical approach. The mean increase in motile sperm/ejaculate after varicocelectomy without testicular delivery was significant (P <0.05) for grade II and III varicoceles. When the testis was delivered, the increase was significant only for grade III varicoceles. No statistically significant difference was found in the change in motile sperm/ejaculate between the two cohorts for all three varicocele grades. The overall pregnancy rate was also not significantly different between the two groups. The testosterone levels rose from 323 to 471 ng/dL on average (P <0.05) after varicocelectomy without testicular delivery but did not change in men who had testicular delivery. The change in testosterone level, age, and testicular volume did not correlate with either the increase in semen parameters or the grade of varicocele treated.

CONCLUSIONS

The results of our study have shown that varicocelectomy without testicular delivery has equivalent or more beneficial effects on semen parameters without affecting varicocele recurrence rates. Delivery of the testis did not offer any beneficial effects on semen quality or pregnancy rates after varicocelectomy.

OBJECTIVE

We evaluated the operative time of microdissection testicular sperm extraction in successful and failed procedures to identify the chance of sperm retrieval during longer microsurgical procedures.

METHODS

A total of 793 men with nonobstructive azoospermia underwent a first attempt at microdissection testicular sperm extraction from January 2000 to September 2009. Clinical factors were analyzed, including age, testicular volume, endocrinological data and histology. Operative time was calculated from incision until the procedure was terminated.

RESULTS

Testicular sperm were successfully retrieved in 57% of the men. Sperm were found within 2, 2 to 4 and 4 to 7 hours in 89%, 30% and 37% of the men, respectively. There were no differences in preoperative clinical characteristics, age, follicle-stimulating hormone, testicular volume, incidence of a Klinefelter's syndrome diagnosis and distribution of most advanced histopathology in patients in the 3 operative time groups. In men in whom sperm were retrieved the clinical pregnancy and live birth rates were 48%, 45% and 29%, and 37%, 30% and 29% for operative times up to 2, 2 to 4 and 4 to 7 hours, respectively (p >0.05). ROC curve analysis of the different operative times for detecting sperm showed that 125 minutes was the most accurate time (AUC 0.81) with 84% sensitivity and 95% specificity.

CONCLUSIONS

The chance of sperm retrieval during microdissection testicular sperm extraction was best during the first 2 hours of the operation. However, sperm were still found in up to 37% of men who required greater than 4 hours of microdissection. Retrospective analysis of our data indicated no cutoff point after which sperm retrieval was uniformly unsuccessful.

OBJECTIVE

Although microdissection testicular sperm extraction has become first line therapy for sperm retrieval in men with nonobstructive azoospermia, there are challenges to the procedure, including difficulty differentiating between seminiferous tubules with normal and abnormal spermatogenesis. Multiphoton microscopy illuminates tissue with a near infrared laser to elicit autofluorescence, which enables real-time imaging of unprocessed tissue without labels. We hypothesized that we could accurately characterize seminiferous tubular histology in humans using multiphoton microscopy.

METHODS

Seven men with normal or abnormal spermatogenesis underwent testicular biopsies, which were imaged by multiphoton microscopy. We assessed these images in blinded fashion. The diagnosis rendered with multiphoton microscopy was then correlated with that of hematoxylin and eosin stained tissue. We evaluated the ability of multiphoton microscopy to differentiate normal from abnormal seminiferous tubules by examining autofluorescence characteristics and diameters, as imaged by multiphoton microscopy. Assessment was repeated with stained slides and results were compared.

RESULTS

The overall concordance rate between multiphoton microscopy and stained slides was 86%. The seminiferous tubules of patients with nonobstructive azoospermia were smaller than those of controls when measured by multiphoton microscopy and staining (p <0.05). The proportion of normal tubules and the diameters obtained with multiphoton microscopy were not different from those obtained with hematoxylin and eosin (p >0.05).

CONCLUSIONS

Multiphoton microscopy can be used to differentiate normal from abnormal spermatogenesis. Its characterization of seminiferous tubular architecture is similar to that provided by hematoxylin and eosin staining. Further investigation of the clinical applications of multiphoton microscopy may improve surgical sperm retrieval outcomes for patients with nonobstructive azoospermia.

Azoospermia in men requires microsurgical reconstruction or a procedure for sperm retrieval with assisted reproduction to allow fertility. While the chance of successful retrieval of sperm in men with obstructive azoospermia approaches >90%, the chances of sperm retrieval in men with non-obstructive azoospermia (NOA) are not as high. Conventional procedures such as fine needle aspiration of the testis, testicular biopsy and testicular sperm extraction are successful in 20-45% of men with NOA. With microdissection testicular sperm extraction (micro-TESE), the chance of successful retrieval can be up to 60%. Despite this increased success, the ability to counsel patients preoperatively on their probability of successful sperm retrieval has remained challenging. A combination of variables such as age, serum FSH and inhibin B levels, testicular size, genetic analysis, history of Klinefelter syndrome, history of cryptorchidism or varicocele and histopathology on diagnostic biopsy have provided some insight into the chance of successful sperm retrieval in men with NOA. The goal of this review was to evaluate the preoperative factors that are currently available to predict the outcome for success with micro-TESE.

There are several options for the treatment of varicocele, including surgical repair either by open or microsurgical approach, laparoscopy, or through percutaneous embolization of the internal spermatic vein. The ultimate goal of varicocele treatment relies on the occlusion of the dilated veins that drain the testis. Percutaneous embolization offers a rapid recovery and can be successfully accomplished in approximately 90% of attempts. However, the technique demands interventional radiologic expertise and has potential serious complications, including vascular perforation, coil migration, and thrombosis of pampiniform plexus. This review discusses the common indications, relative contraindications, technical details, and risks associated with percutaneous embolization of varicocele.

OBJECTIVE

Microdissection testicular sperm extraction has replaced conventional testis biopsies for men with nonobstructive azoospermia and it has become first line treatment. The current problem is that the decision to retrieve tubules is based only on appearance and there is no guarantee that the tubules removed contain sperm. Multiphoton microscopy enables label-free immediate visualization of many biological processes in living tissue at subcellular resolution.

METHODS

We used multiphoton microscopy to study the different developmental stages of spermatogenesis using neonatal, pubertal and adult rat testes. We used a testis hypothermia plus ischemia model to study different testicular histopathologies with multiphoton microscopy. To assess the risk of photo damage DNA fragmentation in testis biopsies imaged at different intensities was assessed by TUNEL assay.

RESULTS

Multiphoton microscopy identified the stage of spermatogenesis in a seminiferous tubule in fresh tissue without using exogenous labels. We noted significant differences in fluorescence and spectroscopic characteristics between tubules with and without sperm. Sertoli's-cell only tubules had abundant autofluorescence in the 420 to 490 and 550 to 650 nm wavelength ranges while tubules containing sperm had autofluorescence only in the 420 to 490 nm range. On DNA fragmentation assay sperm from tubules imaged by multiphoton microscopy had minimal DNA fragmentation at the laser intensities needed to distinguish tubules with and without sperm.

CONCLUSIONS

Multiphoton microscopy has the potential to facilitate real-time visualization of spermatogenesis in humans and aid in clinical applications, such as testicular sperm extraction for men with infertility.

BACKGROUND

Microdissection testicular sperm extraction (micro-TESE) has replaced conventional testis biopsies as a method of choice for obtaining sperm for in vitro fertilization for men with nonobstructive azoospermia. A technical challenge of micro-TESE is that the low magnification inspection of the tubules with a surgical microscope is insufficient to definitively identify sperm-containing tubules, necessitating tissue removal and cytologic assessment. Full field optical coherence tomography (FFOCT) uses white light interference microscopy to generate quick high-resolution tomographic images of fresh (unprocessed and unstained) tissue. Furthermore, by using a nonlaser safe light source (150 W halogen lamp) for tissue illumination, it ensures that the sperm extracted for in vitro fertilization are not photo-damaged or mutagenized.

METHODS

A focal Sertoli-cell only rodent model was created with busulfan injection in adult rats. Ex vivo testicular tissues from both normal and busulfan-treated rats were imaged with a commercial modified FFOCT system, Light-CT™, and the images were correlated with gold standard hematoxylin and eosin staining.

RESULTS

Light-CT™ identified spermatogenesis within the seminiferous tubules in freshly excised testicular tissue, without the use of exogenous contrast or fixation. Normal adult rats exhibited tubules with uniform size and shape (diameter 328 ±11 μm). The busulfan-treated animals showed marked heterogeneity in tubular size and shape (diameter 178 ± 35 μm) and only 10% contained sperm within the lumen.

CONCLUSIONS

FFOCT has the potential to facilitate real-time visualization of spermatogenesis in humans, and aid in micro-TESE for men with infertility.