Isolation of ejaculated spermatozoa six hours after unsuccessful testicular biopsy in nonobstructive azoospermia

Azoospermia, the absence of sperm in ejaculated semen, is the most severe form of male factor infertility and is present in approximately 5% of all investigated infertile couples. The condition is currently classified as obstructive and nonobstructive subgroups. In nonobstructive azoospermia, testicular sperm extraction (TESE) is usually necessary for sperm recovery. This is a case report of pregnancy and subsequent birth of healthy babies following intracytoplasmic sperm injection (ICSI) of ejaculated motile spermatozoa presented 6 h after unsuccessful testicular biopsies in four subsequent in vitro fertilization (IVF) cycles with infertility due to nonobstructive azoospermia.     

Percutaneous aspiration biopsy using an intravenous catheter for testicular sperm retrieval in patients with obstructive azoospermia

Testicular needle aspiration has been proposed as a simple alternative to open biopsy for the diagnosis and treatment of azoospermia. This study describes a new modification of needle aspiration biopsy using an ordinary intravenous catheter to retrieve testicular spermatozoa from patients with obstructive azoospermia, and compares it to the classical fine-needle aspiration (FNA) method. The study included 86 consecutive patients with obstructive azoospermia. Thirty-one patients underwent FNA using butterfly needles (G21 or G23) and a 20 ml syringe. For the remaining 55 patients, an i.v. catheter (G14 or G16) was used instead of the butterfly needle. When testicular tissue was seen in the aspirate, the catheter was clamped and removed from the testis. If, after a maximum of three punctures, insufficient spermatozoa were retrieved, an open biopsy was carried out. Successful sperm retrieval was achieved in 54 out of 55 patients (98.1%) using an i.v. catheter, compared with 16 out of 31 patients (51.6%) using FNA (P < 0.05). Enough aspirated tissue was obtained for cryopreservation of spermatozoa in all 54 patients using the i.v. catheter, compared with 6/16 (37.5%) of the patients undergoing classical FNA. No significant complications were reported. In conclusion, the use of an i.v. catheter for testicular aspiration biopsy significantly improved sperm retrieval compared with FNA in obstructive azoospermia, and allowed for cryopreservation of excess tissue. The procedure is simple and inexpensive, however regular follow up and testing of possible adverse consequences of this method is indicated to establish its safety.     

FSH levels and testicular volumes are associated with the severity of testicular histopathology in men with non-obstructive azoospermia

Purpose

The purpose of this study is to assess a potential association between FSH levels and testicular volumes with the severity of testicular histopathology on testicular biopsy in men with non-obstructive azoospermia (NOA) undergoing microdissection testicular sperm extraction (microTESE).

Methods

A retrospective chart review was performed from the electronic health records of men who underwent microTESE with NOA.

Results

Eighty-six men with NOA underwent microTESE with concomitant testicular biopsy for permanent section to assess the testicular cellular architecture. The histopathological patterns were categorized by severity indicating the odds of sperm retrieval into 2 categories. The unfavorable category included Sertoli cell only pattern and early maturation arrest (n?=?50) and the favorable category included late maturation arrest and hypospermatogenesis patterns (n?=?36). In the men with unfavorable histopathologic patterns, the mean FSH level was 22.9?±?16.6 IU/L, and the mean testicular volume was 10.4?±?6.0 cc. This was in comparison to men with favorable histopathologic patterns revealing a mean FSH level of FSH 13.3?±?12.0 with a mean testicular volume of 13.3?±?5.9 cc. There was a statistically significant higher FSH level in men with unfavorable histopathology than favorable (p?=?0.004) as well as a significant smaller mean testicular volume in men with unfavorable histopathology (p?=?0.029).

Conclusions

Higher serum FSH levels and smaller testicular volumes are associated with more severe testicular histopathological patterns in men with NOA.

     

Intracytoplasmic sperm injection with testicular spermatozoa is less successful in men with nonobstructive azoospermia than in men with obstructive azoospermia

OBJECTIVE: To assess the efficiency of intracytoplasmic sperm injection (ICSI) using testicular spermatozoa in cases of nonobstructive azoospermia. DESIGN: Retrospective case series. SETTING: Tertiary university-based infertility center. PATIENT(S): Overall, 595 couples were included. In 360 couples, the man had normal spermatogenesis. In 118, 85, and 32 couples the man had germ-cell aplasia, maturation arrest, and tubular sclerosis/atrophy, all with focal spermatogenesis present. INTERVENTION(S): We performed 911 ICSI cycles using fresh sperm obtained after testicular biopsies: 306 ICSI cycles used testicular sperm from men with nonobstructive azoospermia, and 605 ICSI cycles used testicular sperm from men with obstructive azoospermia. MAIN OUTCOME MEASURE(S): Fertilization, cleavage, implantation, and pregnancy rates. RESULT(S): Overall, the 2PN fertilization rate was lower in the nonobstructive group: 48.5% vs. 59.7%. There were no differences in in vitro development or in the morphological quality of the embryos. In the nonobstructive group, a total of 718 embryos were transferred (262 transfers) vs. 1,525 embryos in the obstructive group (544 transfers). Both the clinical implantation rate and clinical pregnancy rate per cycle were significantly lower in the nonobstructive group compared with the obstructive group: 8.6% vs. 12.5% and 15.4% vs. 24.0%, respectively. CONCLUSION(S): A statistically significant lower rate of fertilization and pregnancy results from ICSI with testicular sperm from men with nonobstructive azoospermia, compared with men with obstructive azoospermia.     

Cumulative pregnancy rate after ICSI with cryopreserved testicular tissue in non-obstructive azoospermia

The aim of the present study was to describe a simplified and inexpensive method of testicular tissue freezing, to assess the cumulative clinical pregnancy rate (CPR) by this technique, and to provide useful information for counselling couples with non-obstructive azoospermia. One hundred and sixty-five couples with non-obstructive azoospermic males pursuing assisted conception, from December 1995 to December 2002, were included. In all cases, the testicular tissue retrieved by open multiple-biopsy (both sides, by testicular sperm extraction) was frozen using a simple liquid nitrogen vapour freezing technique and was stored in liquid nitrogen thereafter. Only mature spermatozoa were used for intracytoplasmic sperm injection (ICSI) after thawing. Expected CPR were calculated using the Kaplan-Meier survival analysis. A total of 281 cycles were performed resulting in 53 clinical pregnancies. Crude and expected CPR (95% confidence intervals) after three cycles were 32.1 (25.7-40.1) and 55.7% (37.0-74.4) respectively. In conclusion, this simplified method for freezing testicular tissue resulted in a satisfactory outcome after ICSI in cases of non-obstructive azoospermia.     

Large-needle percutaneous aspiration biopsy of the testicle in men with nonobstructive azoospermia

Two hundred thirteen testicular specimens of men with nonobstructive azoospermia were obtained by large-needle percutaneous aspiration biopsy. The mean values of the number of seminiferous tubules per histologic section, dimension, weight, and the fraction of biopsies with spermatogenetic cells (53, 0.2 x 0.3 x 0.62 cm, 385 mg, and 63%, respectively) were similar to those obtained by open or other surgical biopsy techniques that were used for assisted fertilization in the same type of patients.     

Some consequences of vasectomy and testicular biopsy

There has been concern that vasectomy may have an adverse effect on the endocrine function of the testes and that it may cause immunological sensitization of the male to his own spermatozoa. Studies have indicated no deleterious effect on endocrine function, but there is some evidence of potential immunological damage to spermatozoa. Either vasectomy or testicular biopsy may result in immunity to spermatozoa as a result of disturbances to the barrier that sequesters the spermatozoa. Autoallergic orchitis may also occur. Studies have demonstrated that vasectomy or a preexisting state of autoimmunity to spermatozoa during testicular or epididymal surgery, trauma, or infection may precipitate a painful orchitis.     

Value of testicular biopsy in the diagnosis of sterility

Male infertility is a far more prevalent problem than is perhaps generally realized. The evaluation of testicular biopsies allows various clinical syndromes of hypogonadism to be identified and a rational basis for therapy to be established. The morphological-anatomical classification of male infertility paralleling the clinical-biochemical classification divides the different varieties of male infertility into three categories: those due to pretesticular, testicular, and posttesticular causes. Testicular biopsy is an important method in the diagnosis and management of male infertility. The procedure is simple and safe in competent hands and yields information obtainable in no other way. The increasing use of this procedure permitts a rational classification of the testicular lesions responsible for infertility and provides an intelligent basis for the institution of corrective measures or the withholding of therapy in cases in which the biopsy indicates a hopeless prognosis for fertility.     

Enzymatic digestion plus mechanical searching improves testicular sperm retrieval in non-obstructive azoospermia cases

BACKGROUND: In non-obstructive azoospermic patients (NOA) besides the mechanical treatment, vital spermatozoa from the tissue obtained from testes by biopsy can be enzymatically prepared. OBJECTIVE: To increase the sperm recovery success of testicular sperm extraction (TESE), suitable for ICSI. STUDY DESIGN: Prospective, clinical study. In 177 consecutive men who presented with clinical and laboratory data indicating NOA, tissue samples were obtained by microdissection TESE method. Initially, mature spermatozoa were searched for by mechanical extraction technique shredding the biopsy fractions. In cases with no spermatozoa was observed after maximum 30 min of initial searching under the inverted microscope, the procedure was then followed by enzymatic digestion using DNAse and collagenase type IV. In cases of at least a mature spermatozoon could be obtained properly, ICSI was performed. RESULTS: Of 177 cases with NOA, conventional mincing method extended with enzymatic treatment yielded successful sperm recovery for ICSI in 102 (57%). Overall in vitro tissue-processing time for patients with sperm recovery failure after 30 min of mechanical searching, was between 80 and 105 min (mean 96+/-9). Cleavage, embryo transfer and clinical pregnancy rates in mechanical plus enzymatic TESE patients were not significantly different from those of only mechanically TESE performed patients (p>0.05). CONCLUSION: Combination of conventional TESE and enzymatic digestion is an effective method to recover spermatozoa suitable for ICSI. The benefit of the mincing combined with enzyme to sperm retrieval for NOA is firstly to shorten the mechanical searching time, thus minimizing further cellular damage as well as exposure to external conditions, and secondly to reduce the number of cases with sperm recovery failures.     

Interest of pentoxifylline in ICSI with frozen-thawed testicular spermatozoa from patients with non-obstructive azoospermia

Selection for intracytoplasmic sperm injection (ICSI) of viable frozen-thawed testicular spermatozoa obtained from patients suffering from non-obstructive azoospermia is very often long, difficult and sometimes impossible. The purpose of this study was to determine if the use of pentoxifylline (PF) could facilitate this selection in stimulating sperm motility. From January 2000 to December 2004, 108 ICSI cycles with non-obstructive azoospermia were performed. From these 108 cycles, in 64 cycles where no motile spermatozoa were observed or when the time search per spermatozoa was above 20 min, 1.5 mmol/l PF was used for 10 min, whereas the 44 other ICSI cycles were performed using spontaneously motile spermatozoa (control group). In all cases, PF either initiated the motility when no motile spermatozoa were observed, or stimulated the motility, reducing dramatically the time search per spermatozoa. The total fertilization rate was 54.2% versus 66.7% in the control group (P < 0.02). Twenty-nine pregnancies out of the 64 PF cycles (45.3% per cycle) occurred, including 20 deliveries of 23 healthy children and eight ongoing pregnancies, whereas 12 pregnancies were obtained in the control group (27.3% per cycle), including nine deliveries of 13 healthy children. In conclusion, in 100% of cycles pentoxifylline allows the selection of viable frozen-thawed testicular spermatozoa with the same outcome after ICSI as that observed with fresh ejaculated spermatozoa.