Artificial oocyte activation with Ca2+ ionophore improves reproductive outcomes in patients with fertilization failure and poor embryo development in previous ICSI cycles.

Research question: Does artificial oocyte activation (AOA) by a calcium ionophore (ionomycin) improve the previous fertilization failure or poor embryo development of intracytoplasmic sperm injection (ICSI) account for male factor infertility or other infertility causes? Design: This retrospective study involved 114 patients receiving ICSI-AOA in Shanghai First Maternity and Infant Hospital with previous ICSI fertilization failure or poor embryo development. The previous ICSI cycles of the same patients without AOA served as the control group. The fertilization rates, cleavage rates, transferable embryo rates and blastocyst formation rates of the two groups were compared. Additionally, the clinical pregnancy, implantation rate and live birth rates were also compared to assess the efficiency and safety of AOA. Furthermore, two subgroup analyses were performed in this study based on the cause of infertility and the reason for AOA. The fertilization rate, embryonic development potential and clinical outcome were compared among groups. Results: Among 114 ICSI-AOA cycles, the fertilization rate, top-quality embryo rate, implantation rate, clinical pregnancy per patient and live birth rate per patient were improved significantly compared with previous ICSI cycles (p<0.05 to P< 0.001), and the miscarriage rate in the AOA group was significantly lower than that of the control group (p<0.001). In the AOA subgroups based on the cause of infertility, the fertilization rates of each subgroup were significantly improved compared with previous control cycles except for the mixed factor infertility subgroup (p<0.05 to p<0.001). In the AOA subgroups based on the reason for AOA, the fertilization rates of each subgroup were significantly increased compared with those in their previous ICSI cycle without AOA (p<0.001); however, there was no significant difference in the top-quality embryo rate. No significant improvement was found in the implantation rates and the clinical pregnancy rate in each subgroup except for the poor embryo development subgroup. In the 114 AOA cycles, 35 healthy infants (21 singletons and 7 twins) were delivered without major congenital birth defects or malformations. Conclusion: This study showed that AOA with the calcium ionophore ionomycin can improve the reproductive outcomes of patients with previous fertilization failure and poor embryo development after ICSI.

Calcium ionophore improves embryonic development and pregnancy outcomes in patients with previous developmental problems in ICSI cycles.

BACKGROUND: Calcium (Ca2+) ionophores are now mainly considered as efficient treatments for fertilization failure. Recently, its application for rescuing poor embryo development was proposed but still non-routine. This study aimed to explore whether Ca2+ ionophore improves embryo development and pregnancy outcomes in patients with poor embryo development in previous intracytoplasmic sperm injection (ICSI) cycles. METHODS: This study included 97 patients undergoing assisted oocyte activation (AOA) with Ca2+ ionophore (calcimycin, A23187) treatment. Preimplantation embryonic development and clinical outcomes were compared between ICSI-AOA cycles (AOA group) and previous ICSI cycles of the same patients in which poor embryo developmental potential was present (non-AOA group). Subgroups stratified by maternal age (< 35, 35-40, ≥ 40 years, respectively) were analyzed separately. RESULTS: A total of 642 MII oocytes were collected in AOA group, and 689 in non-AOA group. Significantly higher day 3 good quality embryo rate (P = 0.034), good quality blastocyst formation rate (P <  0.001), and utilization rate (P <  0.001) were seen in AOA group. Similar results were seen in each subgroup. For pregnancy outcomes, there were significant differences in clinical pregnancy rate (P = 0.039) and live birth rate (P = 0.045) in total group. In subgroup aged < 35 years, biochemical (P = 0.038), clinical (P = 0.041), and ongoing pregnancy rate (P = 0.037) in AOA group were significantly higher than that in non-AOA group. No significant improvement for clinical outcomes for subgroups aged 35-40 and aged ≥40. CONCLUSION: The study suggests that calcimycin could improve preimplantation development and pregnancy outcomes in patients aged < 35 years with embryo developmental problems in previous ICSI cycles.

Comparison of Human Oocyte Activation Between Round-Headed Sperm Injection Followed by Calcium Ionophore Treatment and Normal Sperm Injection in a Patient With Globozoospermia.

Fertilization failure is common in patients with round-headed sperm, a form of globozoospermia. Artificial oocyte activation is able to assist oocyte fertilization after sperm injection in these patients. Comparisons between oocyte fertilization with or without calcium ionophore have been reported in patients with round-headed sperm. However, no comparison has been reported between round-headed sperm injection followed by calcium ionophone activation and normal sperm injection. In this case report, half of oocytes from a patient were injected with her partner's round-headed sperm followed by calcium ionophore activation, and the other half of oocytes were injected with a donor sperm without calcium ionophore activation. The injected oocytes were cultured to examine fertilization, embryo development, and embryonic aneuploidies in the resulting blastocysts. The fertilization rate was lower in round-headed sperm injected oocytes (3/6) than that in donor sperm injected oocytes (5/6), but rates of blastocyst and aneuploidies were similar in the resulting embryos between the two groups. A euploid blastocyst resulted from round-headed sperm injection was transferred, and a healthy baby was delivered. These results indicate that calcium ionophore treatment can assist oocyte activation in patients with round-headed sperm, but its efficiency to activate oocytes is lower than that induced by a normal sperm injection. However, embryo development and chromosome integrity may not be affected by calcium ionophore treatment.

Assisted oocyte activation with calcium ionophore 44 hours after intracytoplasmic sperm injection resulting in successful pregnancy.

Assisted Oocyte Activation (AOA) with Calcium Ionophore, is possible to manually activate the oocytes and cure globozoospermia, thus leading to successful pregnancy in 1 h after ICSI. But in this case, we report a case that 44 h after ICSI, the arrest zygotes assisted oocyte activation with calcium ionophore, obtained clinical pregnancy and live birth. Accordingly, AOA may provide us with an immediate treatment for embryonic arrest in the future.

Artificial oocyte activation to improve reproductive outcomes in couples with various causes of infertility: a retrospective cohort study.

RESEARCH QUESTION: Does calcium ionophore treatment of oocytes improve fertilization rate, embryo development and outcomes in specific groups of infertile couples? DESIGN: This retrospective cohort study involved 796 couples undergoing oocyte activation with calcium ionophore (A23187) after intracytoplasmic sperm injection (ICSI) between 2016 and 2018. All metaphase II oocytes were exposed to 5 µmol/l ionophore for 15 min immediately after ICSI, cultured in vitro to the blastocyst stage, and transferred to the uteri of recipients on day 5 or cryopreserved for transfer in the next cycle. The previous cycles of the same patients formed the control group. RESULTS: Among 1261 ICSI cycles and 796 ICSI-artificial oocyte activation (ICSI-AOA) cycles, implantation, positive beta-HCG, clinical pregnancy and live birth rates were significantly (P < 0.05 to P < 0.001) improved for all groups, compared with previous cycles, except live birth rate in women with primary ovarian insufficiency (POI). Compared with previous cycles, rates of blastulation (all P < 0.001) and high-quality blastocysts (P < 0.05 to P < 0.001) were increased significantly for couples with male factor (oligoasthenoteratozoospermia [OAT]), unexplained infertility and couples with both factors in the ICSI-AOA cycles. High-quality blastocyst rate was increased in couples with polycystic ovary syndrome (PCOS) (P = 0.0453). Miscarriage rates were decreased significantly (P < 0.05 to P < 0.001) in couples with OAT, PCOS and unexplained infertility in the treatment cycles. No significant differences were found for fertilization rate, embryo development or live birth rate in patients with POI between both groups. CONCLUSIONS: Artificial oocyte activation was able to 'rescue' the poor reproductive outcomes in certain types of infertile couples with history of failure to achieve pregnancy.

Pregnancy and neonatal outcomes of artificial oocyte activation in patients undergoing frozen-thawed embryo transfer: a 6-year population-based retrospective study.

PURPOSE: To evaluate the impact of artificial oocyte activation (AOA) in pregnancy and neonatal outcomes in infertile patients undergoing cryopreserved embryo transfer. METHOD: This retrospective study included 5686 patients' transferred embryos from routine intracytoplasmic sperm injection (ICSI) and 194 patients' transferred embryos from ICSI combined with AOA (ICSI-AOA) from January 2011 to December 2016. Pregnancy and neonatal outcomes of couples undergoing routine ICSI or ICSI-AOA were analyzed before and after propensity score matching. Artificial oocyte activation was performed with ionomycin. RESULTS: The pregnancy outcomes showed no significant difference in the rates of biochemical pregnancy, clinical pregnancy, implantation, miscarriage, ectopic pregnancy, multiple pregnancy, and live births between the routine ICSI and ICSI-AOA groups before and after propensity score matching, respectively. The assessment of neonatal outcomes showed no statistically significant differences in the birth defect rate, birth weight, gestational age, preterm birth rate, early-neonatal death rate, and fetal sex ratio between the two groups, and similar results were also observed in the two matched cohorts. CONCLUSION: Artificial oocyte activation with ionomycin does not adversely affect pregnancy and neonatal outcomes in patients undergoing frozen-thawed embryo transfer, which is beneficial to clinicians counseling patients on the risks of artificial oocyte activation.

Calcimycin Suppresses S100A4 Expression and Inhibits the Stimulatory Effect of Transforming Growth Factor ß1 on Keloid Fibroblasts.

Recent researches have indicated that S100A4 participates in tissue fibrosis, whereas calcimycin inhibits this process as a novel S100A4 transcription inhibitor. However, the relationship and mechanisms between calcimycin and S100A4 in keloid fibroblasts (KFs) remain unknown. The present research was aimed to evaluate the effect of calcimycin on S100A4 expression and pathogenesis in KFs. Keloid fibroblasts were cultured and exposed to different concentrations of calcimycin in the absence or presence of transforming growth factor ß1 (TGF-ß1). The results showed that the expression of S100A4 was significantly increased in keloid derived fibroblasts compared with normal skin fibroblasts. Calcimycin depressed S100A4 in a concentration- and time-dependent manner. Moreover, calcimycin suppressed TGF-ß1-induced collagen type I, fibronectin, and α-smooth muscle actin expression and cell viability in cultured KFs. Furthermore, calcimycin modulated expression of TGF-ß/Smad target genes Smad7 and phosphorylation of TGF-ß1-induced Smad2/3. This research for the first time confirmed the presence of S100A4 in KFs. Calcimycin inhibits the expression of S100A4, as well as KF proliferation and migration and extracellular matrix (ECM) synthesis. Taken together, these results indicate that calcimycin might be a therapeutic candidate to keloid or other related fibrotic disorders.

Unicca(R): nueva superficie de implantes para acelerar la oseointegración / Unicca(R): new surface implants to accelerate the osseointegration

La superficie de los implantes dentales es un parámetro fundamental a considerar para mejorar la regeneración ósea basada en implantes. Junto con las técnicas quirúrgicas y los materiales y herramientas empleados, las modificaciones superficiales de los implantes han ido evolucionando con el tiempo para permitir acortar los tiempos de tratamiento y abordar reconstrucciones cada vez más complejas. La superficie unicCa® resulta de la incorporación a la superficie multirrugosa optima® de una capa de iones de calcio. Las modificaciones realizadas en el desarrollo de esta nueva superficie hacen que presente ventajas como: impedimento del envejecimiento del óxido de titanio, mejora de la unión hueso-implante y aceleración de la fase de la osteointegración. Para demostrar las propiedades de esta nueva superficie se realizaron estudios en cóndilo femoral de conejo y en tibia de oveja. En ambos casos se demostró un incremento y aceleración de la osteointegración. El calcio, presente en la superficie unicCa®, asegura la estimulación celular desde los primeros momentos tras la implantación hasta la consolidación de los tejidos y la formación de la capa calcificada de osteointegración de la que es el constituyente principal. Esto implica una regeneración peri-implante más rápida y de mejor calidad

The surface of the dental implants is a fundamental parameter to consider for improving bone regeneration based on implants. Together with the surgical techniques and materials and tools used, surface modifications of the implants have evolved over time to allow to shorten treatment times and address increasingly complex reconstructions. The unicCa® surface is the result of the addition to the optima® surface of a layer of calcium ions. The modifications made in the development of this new surface make this advantages such as: impairment of the aging of the titanium oxide, improvement of the union bone-implant and acceleration of the phase of the osseointegration. To demonstrate the properties of this new area studies were performed in rabbit femoral condyle and in tibia of sheep. In both cases showed an increase and acceleration of the osseointegration. The calcium, present on the the unicCa® surface ensures the cellular stimulation from the first moments after implantation until the consolidation of the tissues and the formation of the osseointegration of calcified layer which is the main constituent. This implies a regeneration peri-implant faster and of better quality

Does the use of calcium ionophore during artificial oocyte activation demonstrate an effect on pregnancy rate? A meta-analysis.

OBJECTIVE: To study the effect, if any, of calcium ionophore as a method of artificial oocyte activation (AOA) on pregnancy outcomes and fertilization rates. DESIGN: Meta-analysis of randomized controlled trials, prospective observational and retrospective trials, case reports, and a case-control trial. SETTING: University-affiliated teaching hospital. PATIENT(S): Infertile couples undergoing fertilization treatment. INTERVENTION(S): Use of calcium ionophore during AOA. MAIN OUTCOME MEASURE(S): Odds ratio (OR) as the summary statistic for binary variables was used. Both a fixed and random effects model were applied. Subgroup analysis using quantitative methodology (risk of bias, metaregression) and graphical comparison (funnel plot) assessed statistical heterogeneity. RESULT(S): Fourteen studies were selected. AOA with calcium ionophore increased the overall clinical pregnancy rate (per ET; OR = 3.48; 95% confidence interval [CI], 1.65-7.37) and the live birth rate (OR = 3.33; 95% CI, 1.50-7.39). This effect of adding calcium ionophore was further demonstrated with fertilization, cleavage, blastocyst, and implantation rates. Subgroup analysis further supported our findings (studies where n > 10 in both arms; random and fixed effects models). A metaregression (beta = -.145) found that as the quality of the study increases, the effect of calcium ionophore is significantly more pronounced with regards to overall pregnancy rate. CONCLUSION(S): AOA with calcium ionophore treatment after intracytoplasmic sperm injection (ICSI) results in a statistically significant improvement in fertilization, cleavage, blastulation, and implantation rates, as well as overall pregnancy and live-birth rates. The conclusion of this systematic review, demonstrating a strong effect of calcium ionophore use, is reassuring and promising, particularly for couples for whom ICSI alone yields poor fertilization rates.

A preliminary report of successful cleavage after calcium ionophore activation at ICSI in cases with previous arrest at the pronuclear stage.

Artificial oocyte activation (AOA) has been previously suggested as a means to overcome the problem of total fertilization failure, which affects about 1-3% of the intracytoplasmic sperm injection (ICSI) cycles. A preliminary study on the application of chemical AOA was conducted using A23187 Ca(2+) ionophore to improve embryonic development in four women with a history of complete fertilization arrest and inability to transit to cleavage stage during previous ICSI trials. Data indicated that activated oocytes resulted in better fertilization, embryonic development and clinical pregnancy in one of the four couples. Therefore, ICSI combined with AOA using Ca(2+) ionophore may be useful in selected patients with cleavage failure, and may help the zygotes to reach more advanced developmental stages.

Can calcium ionophore "use" in patients with diminished ovarian reserve increase fertilization and pregnancy rates? A randomized, controlled study.

OBJECTIVE: To determine whether calcium ionophore solution can improve the fertilization rate in patients with diminished ovarian reserve whose partners have normal sperm parameters. DESIGN: Between January 2014 and August 2014, patients with diminished ovarian reserve were randomized to make artificial oocyte activation with calcium ionophore solution. SETTING: University hospital. PATIENT(S): A total of 296 patients who had diminished ovarian reserve and partners with normal sperm parameters were included in the study. INTERVENTION(S): Metaphase 2 oocytes were treated with calcium ionophore solution (GM508 Cult-Active) for 15 minutes just after intracytoplasmic sperm injection. MAIN OUTCOME MEASURE(S): Fertilization rate, implantation rate, clinical pregnancy rate, ongoing pregnancy rate. RESULT(S): Fertilization, implantation, pregnancy, and ongoing pregnancy rates for the calcium ionophore and control groups were 60.7% and 55.4%, 12.8% and 10.7%, 21% and 12.8%, and 10.9% and 6.1%, respectively. CONCLUSION(S): This is the first prospective, randomized, controlled study to analyze the effect of calcium ionophore solution on fertilization rate in patients with diminished ovarian reserve. We did not observe any differences in fertilization, clinical pregnancy, or ongoing pregnancy rates between the groups. We propose that fertilization ratios could not be increased by artificial oocyte activation via application of calcium ionophore solution in patients with diminished ovarian reserve. CLINICAL TRIAL REGISTRATION NUMBER: NCT02045914.

[Application of calcium ionophore A23187 in ICSI for globozoospermia: A report of 2 cases and review of the literature].

OBJECTIVE: To investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men. METHODS: We collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease. RESULTS: Quality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group. CONCLUSION: Calcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.

Health of children born through artificial oocyte activation: a pilot study.

Artificial oocyte activation (AOA) has shown to improve fertility in severe male infertility following intracytoplasmic sperm insemination (ICSI). However, the effect of AOA on the health status of children has not been studied. This pilot historical cohort study aims to evaluate physical and mental health of 79 and 89 children from 275 and 406 couples undergoing ICSI-AOA using ionomycin and conventional ICSI, respectively. The outcomes assessed were clinical pregnancy, abortion, type of delivery, and health of children (major birth defect, mental and behavior status). No significant differences were observed between the ICSI-AOA and the ICSI groups for these parameters, and the rate of major birth defects were not significantly different between the 2 groups. In this study, AOA has not imposed a greater risk on physical and mental health of children born through AOA, but for such a solid conclusion, further trails with higher number of cases are required and conclusions drawn are limited to this study.

Activación de ovocitos por ionóforos de calcio en tratamientos de reproducción asistida / Calcium ionophore oocyte activation in assisted reproductive technology

CONTEXTO CLÍNICO: Se estima que una de cada seis parejas no logrará un embarazo exitoso luego de un año de búsqueda. En Argentina se realizan aproximadamente 14 mil tratamientos de reproducción asistida de alta complejidad por año. Con una estimación para Latinoamérica de 100 tratamientos por cada millón de habitantes. Durante la fertilización, normalmente, se produce la activación del ovocito debido a la acción de una organela del espermatozoide denominada acrosoma. Esta activación es dependiente de calcio y de una enzima del acrosoma denominada proteín-lipasa-C-zeta (PLC). Cuando el acrosoma se encuentra poco desarrollado o ausente se produce una falla en la activación del ovocito. Algunos estudios han demostrado que el ovocito puede activarse artificialmente durante una fecundación in vitro con ICSI (inyección intracitoplasmática de espermatozoides) mediante distintos estímulos; entre ellos se puede mencionar el estímulo químico con ionóforos de calcio. Por lo expuesto se postula el uso de ionóforos de calcio en contexto de tratamiento de reproducción asistida, con la finalidad de aumentar la efectividad del tratamiento. TECNOLOGÍA: La inducción de la activación del ovocito mediante el ionóforo de calcio se realiza luego de la ICSI. Los ovocitos son incubados en un medio de cultivo en presencia del ionóforo en determinadas condiciones durante 30 minutos. Luego son transferidos al medio de cultivo habitual de la fertilización in vitro. OBJETIVO: Evaluar la evidencia disponible acerca de la eficacia, seguridad y aspectos relacionados a las políticas de cobertura de la activación del ovocito mediante ionóforos de calcio en tratamiento de reproducción asistida. MÉTODOS: Se realizó una búsqueda en las principales bases de datos bibliográficas (incluyendo Medline, Cochrane y CRD), en buscadores genéricos de Internet, agencias de evaluación de tecnologías sanitarias y financiadores de salud. RESULTADOS: Para el presente documento se incluyeron dos ECAs, un estudio de cohortes y dos series de casos. CONCLUSIONES: La calidad de la evidencia encontrada es moderada. Esta tecnología podría ser útil en parejas con infertilidad que presentan un factor masculino severo con alteraciones del acrosoma espermático, como el déficit de proteín-lipasa-c-zeta y la globozoospermia.

Successful pregnancy after SrCl2 oocyte activation in couples with repeated low fertilization rates following calcium ionophore treatment.

This report describes a successful pregnancy and delivery following oocyte activation with strontium chloride (SrCl2) in couples with repeated complete fertilization failure or low fertilization rates even after calcium ionophore treatment. Eight infertile couples who showed complete fertilization failure or low fertilization rates after conventional intracytoplasmic sperm injection (ICSI) and calcium ionophore treatment. When the results of fertilization were not satisfactory in the cycles, the oocytes were artificially activated by SrCl2 for the next attempts. Oocyte activation with SrCl2 significantly increased the fertilization rates, when compared with conventional ICSI or calcium ionophore treatment (61.7% vs. 20.0% or 25.3%, respectively). There was significant increase in the proportions of good-quality cleaved embryos (50.0% vs. 0% or 12.5%, respectively). The rate of surplus embryos that developed to blastocyst stage increased in SrCl2-treated oocytes, when compared with that in ICSI with or without calcium ionophore treatment (25.7% vs. 0% or 9.1%, respectively). Five successful pregnancies were attained after oocyte activation with SrCl2, of which eight healthy children were born. Physical and mental development of the children were normal from birth to 60 months. These results suggest that SrCl2 in treatment should be considered as an effective method for artificial oocyte activation (AOA) to improve fertilization rates and embryo quality in cases with complete fertilization failure or low fertilization rates after calcium ionophore treatment.

Application of a ready-to-use calcium ionophore increases rates of fertilization and pregnancy in severe male factor infertility.

OBJECTIVE: To analyze whether a ready-to-use calcium ionophore improves outcomes, from fertilization to live birth, in patients with severe male factor infertility. DESIGN: Artificial oocyte activation offered to applicable patients over a 20-month period. SETTING: Specialized in vitro fertilization (IVF) centers in Austria and Germany. PATIENT(S): Twenty-nine azoospermic and 37 cryptozoospermic men. INTERVENTION(S): Mature oocytes treated with a ready-to-use Ca(2+)-ionophore (GM508 Cult-Active) immediately after intracytoplasmic sperm injection (ICSI). MAIN OUTCOME MEASURE(S): Rates of fertilization, implantation, clinical pregnancy, and live birth. RESULT(S): Patients had had 88 previous cycles without artificial activation that resulted in a fertilization rate of 34.7%, 79 transfers (89.8%), and 5 pregnancies, which all spontaneously aborted except one. After artificial oocyte activation, the fertilization rate was 56.9%. In terms of fertilization rate, both azoospermic (64.4%) and cryptozoospermic (48.4%) men statistically significantly benefited from use of the ionophore. In 73 transfer cycles, positive ß-human chorionic gonadotropin levels were observed in 34 cases (46.6%) and 29 cycles (39.7%) that ended with a clinical pregnancy. The corresponding implantation rate was 33.3%. Four spontaneous abortions occurred (11.8%), and 32 healthy children were born. CONCLUSION(S): This is the first prospective multicenter study on artificial oocyte activation in severe male factor infertility. Present data indicate that a ready-to-use calcium ionophore can yield high fertilization and pregnancy rates for this particular subgroup. In addition to fertilization failure after ICSI, severe male factor infertility is an additional area for application of artificial oocyte activation.