Effect of a Freezing Medium Containing Melatonin on Markers of Pre-meiotic and Post-meiotic Spermatogonial Stem Cells (SSCs) After Transplantation in an Azoospermia Mouse Model Due to Testicular Torsion.

Spermatogonial stem cells (SSCs) are essential to the initiation of spermatogenesis. Cryopreservation, long-term maintenance, and auto-transplantation of SSCs could be a new treatment for infertility. The aim of this study was to add melatonin to the basic freezing medium and to evaluate its effect on the efficiency of the thawed SSCs after transplantation into the testicles of azoospermic mice. SSCs were isolated from newborn NMRI mice, and the cells were enriched to assess morphological features. The thawed SSCs were evaluated for survival, apoptosis, and ROS level before transplantation, and the proliferation (MVH and ID4) and differentiation (c-Kit, SCP3, TP1, TP2, and Prm1) markers of SSCs were examined using immunofluorescence, western blot, and quantitative real-time polymerase chain reaction (PCR) after transplantation. It was found that the survival rate of SSCs after thawing was significantly higher in the melatonin group compared with the cryopreservation group containing basic freezing medium, and the rate of apoptosis and level of ROS production also decreased significantly in the cryopreservation group with melatonin (p < 0.05). The expression of proliferation and differentiation markers after transplantation was significantly higher in the cryopreservation group with melatonin compared to the cryopreservation group (p < 0.05). The results suggest that adding melatonin to the basic freezing medium can effectively protect the SSCs by increasing the viability and reducing the ROS production and apoptosis and improve the transplantation efficiency of SSCs after cryopreservation, which will provide a significant suggestion for fertility protection in the clinic.

Testicular sperm extraction and intracytoplasmic sperm injection outcome in cancer survivors with no available cryopreserved sperm.

OBJECTIVE: To assess rates of successful testicular sperm retrieval and intracytoplasmic sperm injection (ICSI) outcome in cancer survivors affected by non-obstructive azoospermia (NOA) or retrograde ejaculation (RE)/failure of emission (FOE). METHODS: A retrospective analysis of cancer survivors who did not cryopreserve sperm prior to treatment undergoing testicular sperm extraction (TESE). Non-cancer NOA patients and neurologic RE/FOE were the control group. RESULTS: A total of 97 cancer survivors were offered TESE and 88 (91%) accepted. Sperm was retrieved and cryopreserved in 34/67 patients with NOA (50.7%) and in 21/21 patients affected by RE/FOE (100%). Sperm retrieval rates were similar in the control group (44.9% in NOA and 100% in RE/FOE). The ICSI cumulative pregnancy rate (60%) and live birth rate (40%) per couple in 30 NOA men did not differ from controls (50.0 and 46.5%, respectively; p = 0.399/0.670). The cumulative pregnancy rate (66.7%) and live birth rate (55.6%) in 18 RE/FOE men did not differ from the control group (38.9 and 33.3%, respectively; p = 0.181/0.315). The cancer type and the resulting infertility disorder (NOA or RE/FOE) were not associated with ICSI outcomes. Female partner age was inversely related to the cumulative live birth rate, being fourfold lower (11.5%) in women ≥ 40 years and 48.8% in younger women (p = 0.0037). CONCLUSIONS: The rate of successful TESE and the ICSI outcome in cancer survivors with NOA and RE/FOE is the same as non-cancer azoospermic patients. Female partner age (older than 40 years) was associated with a significant reduction in live birth rates after TESE-ICSI procedures.

[Hernia surgery: minimization of complications by selection of the "correct mesh"]. / Hernienchirurgie: Minimierung der Komplikationen durch das "richtige Netz".

The risk for developing postoperative complications increases with the degree of surgical trauma, an altered wound healing capability of the patient due to comorbidities and environmental conditions and the selection of an inadequate implant material, the latter offering options for improvement at least in some patients. In general a mesh with large pores made of monofilaments provides a reduced surface area and causes less scarring and inflammation than those with small pores and thereby reduces the rate of scar contraction, pain and the challenge to explant a mesh from a scar bed. When placing the mesh in the abdominal cavity an additional surface coating of polypropylene should prevent the formation of a fistula between mesh and bowel. The risk of recurrence mainly depends on the extent of overlap; however, the flexibility of some meshes may increase the technical difficulties of some implants. In cases of bacterial contamination of the wound there is an increased risk for late onset mesh infection and monofilament meshes offer the best option for complete healing by conservative means. An impaired function of the spermatic cord because of the material, apart from the consequences of the surgical trauma, has not been confirmed in experimental and clinical studies. Revision of mesh sites always is a surgical challenge but could be much easier with implants which are visible in computed tomography (CT) or magnetic resonance imaging (MRI) scans.

Effects of chemotherapy and radiotherapy on spermatogenesis in humans.

Treatment of cancer with chemo- or radiotherapy causes reduction of sperm counts often to azoospermic levels that may persist for several years or be permanent. The time course of declines in sperm count can be predicted by the sensitivity of germ cells, with differentiating spermatogonia being most sensitive, and the known kinetics of recovery. Recovery from oligo- or azoospermia is more variable and depends on whether there is killing of stem cells and alteration of the somatic environment that normally supports differentiation of stem cells. Of the cytotoxic therapeutic agents, radiation and most alkylating drugs are the most potent at producing long-term azoospermia. Most of the newer biologic targeted therapies, except those used to target radioisotopes or toxins to cells, seem to have only modest effects, mostly on the endocrine aspects of the male reproductive system; however, their effects when used in combination with cytotoxic agents have not been well studied.

Management of fertility in patients treated for Hodgkin's lymphoma.

The risk of developing premature ovarian failure and azoospermia is a major concern in long-term survivors treated for Hodgkin's lymphoma. Alkylating chemotherapy containing procarbazine and/or cyclophosphamide causes prolonged azoospermia in 90-100% of men and premature ovarian failure in 5-25% of women under the age of 30. The risk of infertility increases with the cumulative dose of alkylating agents and the risk is high after salvage therapy including conditioning and autologous or allogeneic transplantation. The doxorubicin-bleomycin-vinblastine-dacarbazine regimen is associated with a lower risk of gonadal damage; the rate of infertility is less than 10%. The risk of premature ovarian failure is limited after the doxorubicin-bleomycin-vinblastine-dacarbazine regimen. However, age is an important factor; women over 30 years of age are at a much higher risk of ovarian failure. Semen cryopreservation should be routinely offered, especially before initial treatment with bleomycin, etoposide, doxorubicin, cyclophosphamide, vincristine, procarbazine, prednisolone or salvage therapy with high-dose chemotherapy and autologous transplantation. For women with a stable partner, in vitro fertilization for embryo cryopreservation is a routine procedure but can only be offered to a small number of patients and requires a delay in treatment initiation for at least four weeks. Cryopreservation of mature or immature oocytes remains experimental. Ovarian tissue cryopreservation is promising but has so far resulted in only a small number of pregnancies and births. This method, usually involving the removal of an entire ovary, is only proposed before treatment leading to a high risk of infertility. Analogs of LHRH were investigated in order to preserve fertility in women but are not recommended in the absence of studies demonstrating their effectiveness. The risk of secondary infertility should be discussed with patients from the time of the diagnosis and requires multidisciplinary collaboration between hematologists and Assisted Reproductive Techniques (ART) teams.

Concomitant unilateral and synchronous bilateral testis cancer in azoospermic dizygotic twins: differential management of fertility preservation.

OBJECTIVE: To optimize fertility preservation management in unilateral or bilateral testicular cancer. DESIGN: Case series. SETTING: Urology department and reproductive biology laboratory. PATIENT(S): Dizygotic azoospermic twins presenting unilateral and bilateral synchronous testicular tumors. INTERVENTION(S): Testicular sperm extraction (TESE) and orchiectomy. MAIN OUTCOME MEASURE(S): Semen analysis, histologic diagnosis. RESULT(S): No spermatozoa were cryopreserved for the first case, because fertility preservation was proposed after orchiectomy. Spermatozoa were retrieved after TESE for his brother with bilateral tumor. CONCLUSION(S): Clinicians should be aware of the need to recommend sperm banking before treatments may alter spermatogenesis. TESE may be the sole option for fertility preservation in bilateral testicular cancer.

Damage to the spermatic cord by the Lichtenstein and TAPP procedures in a pig model.

BACKGROUND: Mesh implantation is regarded as the standard treatment of inguinal hernias. Obstructive azoospermia induced by mesh implantation is a rare but serious complication. Whether different operative techniques or mesh materials used have an effect on the integrity of the testicle and spermatic cord remains unclear. MATERIALS: In 12 minipigs a bilateral inguinal hernia repair, either open or laparoscopic, was performed using a standard small-pore polypropylene (PP) or large-pore polyvinyliden fluoride (PVDF) mesh. Next to measurement of the testicular size, thermography of the groin and testicle as a parameter for perfusion was performed preoperatively and at a follow-up at 6 months. Obstructions of the vas deferens were estimated radiographically. Testicular function (Johnson score) and mesh integration (granuloma size, apoptotic cells) were analyzed histologically. RESULTS: Mean testicular size did not change significantly in follow-up compared to preoperative values. Technique and mesh material used failed to have a significant influence. Thermography of the groin following the Lichtenstein technique had significantly higher values at follow-up regardless of the mesh used. This could not been shown for laparoscopic treatment. Thermographic measurements at the testicle showed a significantly increased temperature in all groups compared to preoperative measurements. Only the Lichtenstein PP group showed significantly decreased values in testicular function. Quantity and quality of obstructions seen at vasography were most detectable in the Lichtenstein PP group. There was significantly decreased granuloma formation following PVDF mesh implantation compared to the PP mesh group regardless of the technique used. CONCLUSIONS: Both the technique and the mesh material have an impact on integrity of spermatic cord and testicular function. According to the results of this study, the laparoscopic TAPP procedure using a large-pore PVDF mesh has the least effect compared to preoperative values.

Male gonadal toxicity.

Cancer treatment with chemotherapy or radiotherapy causes gonadal toxicity in male patients. The endpoint of most concern for future reproductive options is the induction of prolonged azoospermia, which may or may not be reversible. The immediate effects of therapy and its reversibility are most readily observed in post-pubertal patients, but the same antineoplastic regimens given to prepubertal males can induce permanent azoospermia. The probability of permanent azoospermia is related to the specific agents used and their doses. The most damaging are alkylating agents (particularly chlorambucil, procarbazine, cyclophosphamide, melphalan, and busulfan), cisplatin and radiation to the region of the testicles.

Infertility and bilateral adrenal tumors.

An infertile man who presents with azoospermia and bilateral adrenal tumors reveals an XX karyotype and congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Hysterectomy and bilateral oophorectomy performed in infancy and lack of treatment resulted in full masculinization.