Infertility, MaleNCT05161247
Reproductive Outcome After Sperm Selection for Intracytoplasmic Sperm Injection (ICSI) Using Zona Pellucida Proteins
A study testing Zona pellucida sperm selection, Conventional sperm selection and Intracytoplasmic Sperm Injection (ICSI) in people with Infertility, Male, run by Nadezhda Women's Health Hospital at 1 site in Bulgaria.
Trial 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. Read more about this study
- Who is it for?
- Ages 20 to 50 years
- Conditions the study is for
- Infertility, Male
- What is being tested?
- Zona pellucida sperm selection, Conventional sperm selection and Intracytoplasmic Sperm Injection (ICSI)
- Type of treatment
- Injection or infusion
- Could I receive a placebo?
- No, everyone receives an active treatment
- How many people
- 368 participants
- Where
- 1 siteBulgaria
- Study dates
- 3 Dec 2021to 5 Jul 2024
- Phase
- Interventional
- Are there results?
- None posted in the registry
Key requirements
Who can take part, as listed by the sponsor. Only the study team can confirm whether you are eligible, so take this list to your doctor.
- Ages 20 to 50 years
- All sexes
- No healthy volunteers
- Adult
Who may be able to join
- 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
Who cannot join
- Presence of low quality oocytes
- Females with endometrial or ovarian infertility factor
Trial locations and contacts
1 site in 1 country.
About this study
Official title: 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.
Read the 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.
- Conditions
- Infertility, Male
- Being tested
- Zona pellucida sperm selection, Conventional sperm selection, Intracytoplasmic Sperm Injection (ICSI)
- Keywords
- zona pellucida, sperm selection, ICSI, ART outcomes
- Medical subject headings
- Infertility, Male
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Zona pellucida sperm selectionOther
Sperm selection for ICSI based on the spermatozoa ability to adhere to immobilized zona pellucida proteins.
Conventional sperm selectionOther
Sperm selection for ICSI based on the sperm morphology.
Intracytoplasmic Sperm Injection (ICSI)Other
Normal ICSI procedure.
| Study group | Role | What they receive |
|---|---|---|
| Zona pellucida sperm selection | Experimental | 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. Receives: Other: Zona pellucida sperm selection; Other: Intracytoplasmic Sperm Injection (ICSI) |
| Conventional sperm selection | Active comparator | 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. Receives: Other: Conventional sperm selection; Other: Intracytoplasmic Sperm Injection (ICSI) |
What is being measured
The primary outcome is the main question the study is designed to answer.
Fertilisation rate
Defined as number of fertilized oocytes (presence of 2 pronuclei) per total number of injected oocytes.
Measured: 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.
Measured: 2 to 6 days after ICSI
Implantation rate
Defined as high levels of human chorionic gonadotropin (hCG) in the peripheral blood per transfer.
Measured: 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.
Measured: 6 weeks after embryo transfer
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted17 Dec 2021
- Study start3 Dec 2021
- Primary completion3 Mar 2024
- Study completion5 Jul 2024
Resources
- Official record on ClinicalTrials.govThe registry entry this page is built from, with its full change history.
- Publications (1)Papers the sponsor has linked to this study.
- Other trials for Infertility, MaleStudies of the same condition, open and under way.
- What the trial phases meanHow a treatment moves from first tests in people to approval.
More details
For doctors and researchers: how the study is designed, who runs it, and its registry IDs.
Study design
- Study type
- Interventional
- Phase
- Phase N/A
- Primary purpose
- Treatment
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Model description
- After couples are recruited for the study they will be randomly allocated to zona selection group (study group) and normal ICSI (control group). For the study group zonae pellucidae from germinal vesicle (GV) or metaphase I (MI) stage oocytes from the female partners will be collected and acid-solubilised to obtain zona proteins. The acid-solubilised zonae will be coated on petri dishes. On the day of their follicular punction the spermatozoa for ICSI in the study group will be selected according to their ability to adhere to the coated zona pellucida proteins from their partner. On the day of their follicular punction the spermatozoa for ICSI in the control group will be selected with no intervention.
- Masking
- Single blind
- Who is masked
- Outcomes assessor
- Enrolment
- 368 participants (actual)
Sponsor and investigators
- Lead sponsor
- Nadezhda Women's Health Hospital
- Sponsor type
- University, hospital or other
- Responsible party
- Sponsor
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT05161247
- Sponsor's protocol ID
- 59/03122021
- Status last verified
- May 2025
- First submitted
- 6 Dec 2021
- Last updated
- 21 May 2025
- Results posted
- No
- Data monitoring committee
- No
- FDA-regulated drug
- No
- FDA-regulated device
- No
- Individual data sharing
- No
Publications
Ganeva R, Parvanov D, Vidolova N, Handzhiyska M, Ruseva M, Vasileva M, Nikolova K, Ivanova I, Shaban M, Shabarkova J, Hristova R, Miladinova M, Stamenov G. Sperm selection by zona adhesion improves assisted reproductive treatment outcomes. Andrology. 2024 Sep;12(6):1373-1380. doi: 10.1111/andr.13590. Epub 2024 Jan 15.
PubMed 38225818
Study at a glance
From the registry record, last updated by the sponsor on 21 May 2025. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Assessment of the IVF Outcome Parameters After Sperm Selection by Adhesion to Zona Pellucida Proteins
- ClinicalTrials.gov ID
- NCT05161247
- Acronym / sponsor ID
- 59/03122021
- Conditions
- Infertility, Male
- Keywords
- zona pellucida, sperm selection, ICSI, ART outcomes
- Study type
- Interventional
- Recruitment status
- Completed
- Enrolment
- 368 participants (actual)
- Lead sponsor
- Nadezhda Women's Health HospitalUniversity, hospital or other
- Study start
- 3 Dec 2021 (actual)
- Primary completion
- 3 Mar 2024 (actual)
- Study completion
- 5 Jul 2024 (actual)
- First posted
- 17 Dec 2021 (actual)
- Results
- Not posted in the source record
Clinical summary
The research question as the sponsor registered it. This describes what the study plans to test, not what it found.
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 design
- Primary purpose
- Treatment
- Allocation
- Randomised
- Intervention model
- Parallel groups
- Masking
- Single blind (outcomes assessor)
- Number of arms
- 2
- Model description
- After couples are recruited for the study they will be randomly allocated to zona selection group (study group) and normal ICSI (control group). For the study group zonae pellucidae from germinal vesicle (GV) or metaphase I (MI) stage oocytes from the female partners will be collected and acid-solubilised to obtain zona proteins. The acid-solubilised zonae will be coated on petri dishes. On the day of their follicular punction the spermatozoa for ICSI in the study group will be selected according to their ability to adhere to the coated zona pellucida proteins from their partner. On the day of their follicular punction the spermatozoa for ICSI in the control group will be selected with no intervention.
| Arm | Type | Receives |
|---|---|---|
| Zona pellucida sperm selection | Experimental | Other: Zona pellucida sperm selection, Other: 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 | Active comparator | Other: Conventional sperm selection, Other: 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. |
Who may qualify
- Ages
- 20 to 50 years
- Sex
- All
- Healthy volunteers
- Not accepted
- Age groups
- Adult
Key inclusion criteria (6 listed)
- 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.
Key exclusion criteria (2 listed)
- Presence of low quality oocytes
- Females with endometrial or ovarian infertility factor
Eligibility is decided by the study team after screening; this overview does not establish that a patient qualifies. Full inclusion and exclusion criteria.
Interventions
| Intervention | Type | Arms |
|---|---|---|
| Zona pellucida sperm selectionSperm selection for ICSI based on the spermatozoa ability to adhere to immobilized zona pellucida proteins. | Other | Zona pellucida sperm selection |
| Conventional sperm selectionSperm selection for ICSI based on the sperm morphology. | Other | Conventional sperm selection |
| Intracytoplasmic Sperm Injection (ICSI)Normal ICSI procedure. | Other | Conventional sperm selection, Zona pellucida sperm selection |
Primary outcome
Fertilisation rate
Time frame: 24 hours after ICSI
Defined as number of fertilized oocytes (presence of 2 pronuclei) per total number of injected oocytes.
Embryo quality
Time frame: 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
Time frame: 14 days after embryo transfer
Defined as high levels of human chorionic gonadotropin (hCG) in the peripheral blood per transfer.
Clinical pregnancy rate
Time frame: 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.
Results and safety
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
Research links
- Official record on ClinicalTrials.govLast updated by the sponsor 21 May 2025.
- Derived from this study: PubMed 38225818Ganeva R, Parvanov D, Vidolova N, Handzhiyska M, Ruseva M, Vasileva M, Nikolova K, Ivanova I, Shaban M, Shabarkova J, Hristova R, Miladinova M, Stamenov G. Sperm selection by zona adhesion improves assisted reproductive treatment outcomes. Andrology. 2024 Sep;12(6):1373-1380. doi: 10.1111/andr.13590. Epub 2024 Jan 15.
- Individual participant dataSharing planned: No
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Referral and contact
AzTrials does not enrol participants or take referrals. Contact the study team or a site listed below; they decide who can take part. This study is not recruiting at the moment.
The record lists no contact. Use the official record to reach the sponsor.
1 site in 1 country: see every site and its status.
Questions about this trial
What is this clinical trial studying?
A study testing Zona pellucida sperm selection, Conventional sperm selection and Intracytoplasmic Sperm Injection (ICSI) in people with Infertility, Male, run by Nadezhda Women's Health Hospital at 1 site in Bulgaria.
The full registered description is in the About this study section and the official record.
Who may be eligible to take part?
The sponsor lists 6 inclusion and 2 exclusion criteria. Eligibility is decided by the study team after screening, so read the full criteria and contact a listed site.
Is this clinical trial currently recruiting?
The record says: finished, as of the sponsor's last update on 21 May 2025. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 1 site in Bulgaria. Whether a site is taking part right now should be confirmed with that site. See every site.
What are the possible benefits and risks?
They depend on the study and the treatment, and no study can promise a benefit. The study team explains the known risks during informed consent, before anyone joins; the record itself does not list them.
Are results available for this study?
The study is completed, but results are not available in the source record. That does not mean the treatment did or did not work.
How can a doctor refer a patient or contact the research team?
The record lists no contact. The official record names the sponsor, who can be asked directly. AzTrials does not enrol participants or take referrals.
What does the primary outcome measure?
Fertilisation rate (time frame: 24 hours after ICSI). The study lists 4 primary outcomes in all. It is what the study was designed to measure, not a finding.
Where can I read the original study information?
On the ClinicalTrials.gov record for NCT05161247, which also lists 1 linked publication.
Does a listing mean the treatment is approved or proven to work?
No. A listing means the study is registered. It does not show that the treatment is safe, works, or is approved by a regulator.
How often is this information updated?
AzTrials copies changed records from ClinicalTrials.gov every night and reloads the whole registry every week. The sponsor last updated this record on 21 May 2025.