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

Assessment of the IVF Outcome Parameters After Sperm Selection by Adhesion to Zona Pellucida Proteins

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.

Study Overview

• Status: Completed
• Conditions: Infertility, Male
• Intervention / Treatment: Other: Zona pellucida sperm selection; Other: Intracytoplasmic Sperm Injection (ICSI); Other: Conventional sperm selection

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 Type: Interventional
• Enrollment (Actual): 368
• Phase: Not Applicable

Contacts and Locations

Study Locations

• Bulgaria
  • Sofia, Bulgaria, 1373
    • Nadezhda Women's Health Hospital

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 20 years to 50 years (Adult)
• Accepts Healthy Volunteers: No

Description

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 Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Single

Number of Arms

2

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
Experimental: 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.	Other: Zona pellucida sperm selection; Sperm selection for ICSI based on the spermatozoa ability to adhere to immobilized zona pellucida proteins.; Other: Intracytoplasmic Sperm Injection (ICSI); Normal ICSI procedure.
Active Comparator: 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.	Other: Intracytoplasmic Sperm Injection (ICSI); Normal ICSI procedure.; Other: Conventional sperm selection; Sperm selection for ICSI based on the sperm morphology.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Fertilisation rate	Defined as number of fertilized oocytes (presence of 2 pronuclei) per total number of injected oocytes.	24 hours after ICSI
Embryo quality	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.	2 to 6 days after ICSI
Implantation rate	Defined as high levels of human chorionic gonadotropin (hCG) in the peripheral blood per transfer.	14 days after embryo transfer
Clinical pregnancy rate	Defined as number of gestational sacs with fetal heart beat, shown by ultrasound in gestational week 6 per number of embryo transferred.	6 weeks after embryo transfer

Collaborators and Investigators

• Sponsor: Nadezhda Women's Health Hospital

Study record dates

Study Major Dates

• Study Start (Actual): December 3, 2021
• Primary Completion (Actual): March 3, 2024
• Study Completion (Actual): July 5, 2024

Study Registration Dates

• First Submitted: December 6, 2021
• First Submitted That Met QC Criteria: December 6, 2021
• First Posted (Actual): December 17, 2021

Study Record Updates

• Last Update Posted (Actual): May 21, 2025
• Last Update Submitted That Met QC Criteria: May 20, 2025
• Last Verified: May 1, 2025

More Information

Terms related to this study

• Keywords: ICSI; sperm selection; zona pellucida; ART outcomes
• Additional Relevant MeSH Terms: Urogenital Diseases; Genital Diseases; Genital Diseases, Male; Male Urogenital Diseases; Infertility; Infertility, Male
• Other Study ID Numbers: 59/03122021

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No