Reproductive Outcome After Sperm Selection for Intracytoplasmic Sperm Injection (ICSI) Using Zona Pellucida Proteins

Not Applicable

Completed

• Conditions: Infertility, Male
• Interventions: Other: Zona pellucida sperm selection; Other: Conventional sperm selection; Other: Intracytoplasmic Sperm Injection (ICSI)

• Registration Number: NCT05161247
• Lead Sponsor: Nadezhda Women's Health Hospital

Brief Summary

The aim of this study is to analyze the effect of a sperm selection method by adhesion to partners' zona pellucida proteins on the assisted reproduction treatment (ART) outcome parameters.

Detailed Description

The success rate after treatment by in-vitro fertilisation (IVF) depends on the sperm selection process. Classical sperm selection techniques are based on the spermatozoa morphology and motility, therefore are not able to measure the sperm ability to recognise the oocyte. Furthermore it is known that semen samples showing higher adherence rate to zona pellucida have higher fertilisation potential than spermatozoa that fail to bind to zona proteins.

There have been numerous reports on applying partner' native zona pellucida in sperm selection prior ICSI. However, these studies imply the whole native zona pellucida, while this study will focus on the application of acid-solubilized zonae pellucidae.

Here in this study, the aim is to assess the fertilisation rate, the number and quality of the formed embryos, implantation and ongoing pregnancy success after sperm selection for ICSI by their ability to adhere to immobilized zona pellucida proteins.

Study Timeline

• Study Start: 2021-12-03
• Study First Submitted: 2021-12-06
• Study First Submitted QC: 2021-12-06
• Study First Posted: 2021-12-17
• Primary Completion: 2024-03-03
• Study Completion: 2024-07-05
• Status Verified: 2025-05
• Last Update Submitted: 2025-05-20
• Last Update Posted: 2025-05-21

Recruitment & Eligibility

• Status: COMPLETED
• Sex: All
• Target Recruitment: 368

Inclusion Criteria

• Clinical diagnosis for primary or secondary infertility
• Healthy females
• Clinical diagnosis for unexplained infertility
• Females between 20 and 45 years old.
• Males between 20 and 50 years old.
• At least 3 retrieved oocytes per follicular puncture with at least 1 oocyte at germinal vesicle (GV) or metaphase I (MI) stage

Exclusion Criteria

• Presence of low quality oocytes
• Females with endometrial or ovarian infertility factor

Study & Design

• Study Type: INTERVENTIONAL
• Study Design: PARALLEL

Arm && Interventions

Group	Intervention	Description
Zona pellucida sperm selection	Intracytoplasmic Sperm Injection (ICSI)	Semen processing: liquefaction followed by centrifugation and swim-up. Zona pellucida sperm selection: Spermatozoa are selected according to their ability to adhere to zona pellucida proteins obtained from their partners native zona pellucida. Intracytoplasmic sperm injection (ICSI): Each oocyte is injected with a single morphologically normal spermatozoa. The injection procedure is carried out in a sterilized dish using holding pipette and injection needle.
Conventional sperm selection	Intracytoplasmic Sperm Injection (ICSI)	Semen processing: liquefaction followed by centrifugation and swim-up. Samples are incubated until time of injection. Conventional sperm selection: Spermatozoa are selected according to their morphology. Intracytoplasmic sperm injection (ICSI): Each oocyte is injected with a single morphologically normal spermatozoa. The injection procedure is carried out in a sterilized dish using holding pipette and injection needle.
Zona pellucida sperm selection	Zona pellucida sperm selection	Semen processing: liquefaction followed by centrifugation and swim-up. Zona pellucida sperm selection: Spermatozoa are selected according to their ability to adhere to zona pellucida proteins obtained from their partners native zona pellucida. Intracytoplasmic sperm injection (ICSI): Each oocyte is injected with a single morphologically normal spermatozoa. The injection procedure is carried out in a sterilized dish using holding pipette and injection needle.
Conventional sperm selection	Conventional sperm selection	Semen processing: liquefaction followed by centrifugation and swim-up. Samples are incubated until time of injection. Conventional sperm selection: Spermatozoa are selected according to their morphology. Intracytoplasmic sperm injection (ICSI): Each oocyte is injected with a single morphologically normal spermatozoa. The injection procedure is carried out in a sterilized dish using holding pipette and injection needle.

Primary Outcome Measures

Name	Time	Method
Embryo quality	2 to 6 days after ICSI	Morphological evaluation of the cultured embryos will be conducted 120 or 144 hr following sperm injection. Blastocysts scoring will be performed on day 5 or 6 depending on the blastocoel cavity expansion and on the inner cell mass and trophectoderm cells integrity.
Implantation rate	14 days after embryo transfer	Defined as high levels of human chorionic gonadotropin (hCG) in the peripheral blood per transfer.
Fertilisation rate	24 hours after ICSI	Defined as number of fertilized oocytes (presence of 2 pronuclei) per total number of injected oocytes.
Clinical pregnancy rate	6 weeks after embryo transfer	Defined as number of gestational sacs with fetal heart beat, shown by ultrasound in gestational week 6 per number of embryo transferred.

Trial Locations

Locations (1)

Nadezhda Women's Health Hospital; 🇧🇬 Sofia, Bulgaria