InfertilityNCT02329704
Evaluation Study Between Density Gradient and Swimming Down for Semen Processing
An observational study of Infertility, run by Ahmad Mustafa Mohamed Metwalley at 2 sites in 2 countries.
Trial summary
This study evaluates the quality of sperm produced after semen processing using Density Gradient (DG) and swimming down (SD). Read more about this study
- Who is it for?
- Ages 18 to 60 years, men only
- Conditions the study is for
- Infertility
- What is being tested?
- Diff-Quik staining for morphology evaluation, Motility Evaluation for grade A and B and Concentration (mil/ml)
- Type of treatment
- Surgery or procedure
- How many people
- 100 participants
- Where
- 2 sitesin 2 countries
- Study dates
- May 2007to Sep 2007
- Phase
- Observational
- 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 18 to 60 years
- Men only
- No healthy volunteers
- Adult
Study population: Cases arranged for study were submitted for routine semen analysis. Patient agreements collected as consent form signed during sample submission.
Who may be able to join
- Semen with motility more than 10 % according WHO 2010 standers
- Semen with sperm count more than 5 mil/ml according WHO 2010 standers
Who cannot join
- Semen volume less than 1.0 ml
- Semen with liquefaction time more than 60 min
Trial locations and contacts
2 sites in 2 countries.
2 sites in 2 countries
Egypt1 site
Ibn Sina IVF Center- Ibn Sina Hospital
Sohag, Sohag Governorate, 15006
About this study
This study evaluates the quality of sperm produced after semen processing using Density Gradient (DG) and swimming down (SD).
Read the detailed description
Semen processing to be used for different ART procedures is essential step to isolate dead sperms as well as those abnormally structural or morphology. Semen preparation will maintain to remove the seminal fluids and present cells and contamination pus cells or bacterial cells.
Among different protocols, Density Gradient and swim-up are the most common. For poor motility and number sample Density Gradient is indicated. While final evaluation for produced or separated sperm form processing procedures was not evaluated yet.
Preparation for evaluation study to compare output of main used protocol Density Gradient (DG) with other protocol as swimming down using different study criteria can make more clear evaluation for better processing procedures.
The cases arranged for evaluation were semen samples are within WHO 2010 normal values for motility and count to be more than 5 million/ml. All samples to be divided to two portions for processing by the two preparation techniques.
Post preparation evaluation to be done for the two outputs using: Motility monitoring for 48 hours at 37.0 Dc, abnormal forms, hyaluronan affinity testing and contaminating bacterial growth.
In this study we will show drawback effect of physical and time stress interface applied on sperms during processing, and compare the out put results with that results produced from sperm processed without centrifugation interface or more processing time. As the sperm will separate it self by its natural movement and normal behavior.
Evaluation will be on three categories:
- The final output quality: as net highly progressive sperms and their count number ( sperm morphology, count and motility): as our results showed more phonologically normal sperm retrieved from SD method with higher count and more percentage of progressively motile sperms.
- The quality of sperm produced: through monitoring sperm vitality testing or sperm motility power among 2 days at 37.0 Ds. Motility will be evaluated after 24 hours and 48 hours. Both readings showed more decrease in sperm motility and vitality with time consuming with portions processed using density gradient. While portions processed using SD showed significant better readings. Another functional activity of sperm post processing is using HBA testing slide, as we evaluate the sperm affinity to hyaluronan. This Testing or assay indicate the physiological activity of sperm still maintained. While it showed HBA higher significant results with swimming down group study.
- Final evaluation, that are used to present the quality of separation: is monitored using the contamination and carry over effect during semen manipulation and processing. Bacterial contamination using routine culture media (Blood agar) to evaluate 5 microns from each portions for presence and absence of any bacterial cells or during processing and manipulation.
- Conditions
- Infertility
- Being tested
- Diff-Quik staining for morphology evaluation, Motility Evaluation for grade A and B, Concentration (mil/ml), 24 H half life, 48 H half life
- Keywords
- IUI/AIH, Semen preparation, Semen processing, ICSI/IVF, ART, Density Gradient, Swimming down, hyaluronan affinity testing, Sperm vitality testin
- Medical subject headings
- Infertility
What participants receive
What participants receive. In a randomised study each person is assigned to one group by chance.
Diff-Quik staining for morphology evaluationProcedure or surgery
Also called Abnormal forms %
Papanicolaou stain (haematoxylin, orange G6, and EA50) gives a clear difference between basophilic and acidophilic cell constituents and thereby enables a detailed examination of chromatin pattern, which is useful in the evaluation of sperm morphology and assessment of presence of immature spermatozoa. Cytoplasmic staining can vary between red and green dependent on ionic strength, pH, and composition of the cell department and the stain (OG6 and EA50).
Morphological assessment for processed portions of semen. As separation process has to minimize the morphological abnormal sperm.
Motility Evaluation for grade A and BProcedure or surgery
Also called Motility %
The better separation method will insure more progressive sperms after processing.
Grading using A and B scale scored by (WHO 2010) references.
Concentration (mil/ml)Procedure or surgery
Also called Concentration for final output
count of isolated sperms Using Makler chamber for count of sperm for each portion post processing.
24 H half lifeProcedure or surgery
Also called Motility after 24 hours incubation at 37 Dc.
percentage of active motile to non motile sperms.
Microscopic examination for each sample after 24 hours incubation, to evaluate the progressively motile sperm and those non progressive or non motile.
48 H half lifeProcedure or surgery
Also called Motility evaluation after 48hours incubation at 37 Dc.
percentage of active motile to non motile sperms.
Microscopic examination for each sample after 48 hours incubation, to evaluate the progressively motile sperm and those non progressive or non motile.
HBA TestingProcedure or surgery
Also called Sperm physioogical activity evaluation HA binding
The Hyaluronic Binding Assay (HBA®) is an important diagnostic tool for suspected male infertility in the analysis of semen. In a matter of minutes the HBA® slide provides an answer to the proportion of mature binding spermatozoa in the sample (The HBA® score %).
By using HBA slide we can count the physiologically active sperms at each sample and calculate percentage of active sperms with reference to post processing count from 3rd investigating arm.
Bacterial contaminationBiological
Also called Bacterial contamination presence of post processinf samples
Routine bacterial culture using Blood Agar culture plates for 10 micron finally washed samples, to confirm presence of any bacteria contamination caused by carry over during processing.
| Study group | Role | What they receive |
|---|---|---|
| Density Gradient processing | Semen portion processed using Density Gradient (80/40 %) Spin on 1200 r.p.m./20min Wash on 1200 r.p.m./5min Rewash on 1200 r.p.m./5min
then evaluation to be done using : Diff-Quik staining for morphology evaluation Motility Evaluation for grade A and B Concentration (mil/ml) 24 H half life 48 H half life HBA Testing Bacterial contamination Receives: Procedure: Diff-Quik staining for morphology evaluation; Procedure: Motility Evaluation for grade A and B; Procedure: Concentration (mil/ml); Procedure: 24 H half life; Procedure: 48 H half life; Procedure: HBA Testing; Biological: Bacterial contamination | |
| Swimming down Processing | Semen portion processed using swimming down (100 %) 30 min at room temperature processing for swim down Wash on 1200 r.p.m./5min Rewash on 1200 r.p.m./5min
then evaluation to be done using : Diff-Quik staining for morphology evaluation Motility Evaluation for grade A and B Concentration (mil/ml) 24 H half life 48 H half life HBA Testing Bacterial contamination Receives: Procedure: Diff-Quik staining for morphology evaluation; Procedure: Motility Evaluation for grade A and B; Procedure: Concentration (mil/ml); Procedure: 24 H half life; Procedure: 48 H half life; Procedure: HBA Testing; Biological: Bacterial contamination |
What is being measured
The primary outcome is the main question the study is designed to answer.
HBA
Affinity is measured according to percentage of non motile sperm produced from 10 µL of processed sample.
Measured: 1 day
Abnormal forms
Percentage normally stained sperms after processing
Measured: 1 day
Secondary outcomes (3)
Final concentration
Concentration was fluctuation according to original motility and concentration, DG
Measured: 1 day
24 Hour motility
Motility evaluation after 37.0 Dc incubation
Measured: 1 day
48 Hour motility
Motility evaluation after 37.0 Dc incubation
Measured: 2 days
Timeline
From the start of the study to its completion; the mark is today. Estimated dates are the sponsor’s plan.
- First posted1 Jan 2015 (estimated)
- Study startMay 2007
- Primary completionSep 2007
- Study completionSep 2007
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 InfertilityStudies 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
- Observational
- Observational model
- Case-control
- Time perspective
- Prospective
- Enrolment
- 100 participants (actual)
Sponsor and investigators
- Lead sponsor
- Ahmad Mustafa Mohamed Metwalley
- Sponsor type
- University, hospital or other
- Responsible party
- Sponsor-investigator, Ahmad Mustafa Mohamed Metwalley, Al Baraka Fertility Hospital
- Investigators
- Ahmad M Metwalley, Study director, Adliyah
IDs, dates and publications
- ClinicalTrials.gov ID
- NCT02329704
- Sponsor's protocol ID
- AlBarakaSD2
- Status last verified
- Dec 2014
- First submitted
- 28 Dec 2014
- Last updated
- 1 Jan 2015
- Results posted
- No
- Data monitoring committee
- No
Publications
Malvezzi H, Sharma R, Agarwal A, Abuzenadah AM, Abu-Elmagd M. Sperm quality after density gradient centrifugation with three commercially available media: a controlled trial. Reprod Biol Endocrinol. 2014 Dec 2;12:121. doi: 10.1186/1477-7827-12-121.
PubMed 25466430
Study at a glance
From the registry record, last updated by the sponsor on 1 Jan 2015. Recruitment status is as the sponsor reported it: confirm with the study team.
- Official title
- Evaluation Study Between Density Gradient and Swimming Down for Semen Processing
- ClinicalTrials.gov ID
- NCT02329704
- Acronym / sponsor ID
- AlBarakaSD2
- Conditions
- Infertility
- Keywords
- IUI/AIH, Semen preparation, Semen processing, ICSI/IVF, ART, Density Gradient, Swimming down, hyaluronan affinity testing
- Study type
- Observational
- Recruitment status
- Completed
- Enrolment
- 100 participants (actual)
- Lead sponsor
- Ahmad Mustafa Mohamed MetwalleyUniversity, hospital or other
- Study start
- May 2007
- Primary completion
- Sep 2007 (actual)
- Study completion
- Sep 2007 (actual)
- First posted
- 1 Jan 2015 (estimated)
- 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.
This study evaluates the quality of sperm produced after semen processing using Density Gradient (DG) and swimming down (SD).
Detailed description
Semen processing to be used for different ART procedures is essential step to isolate dead sperms as well as those abnormally structural or morphology. Semen preparation will maintain to remove the seminal fluids and present cells and contamination pus cells or bacterial cells.
Among different protocols, Density Gradient and swim-up are the most common. For poor motility and number sample Density Gradient is indicated. While final evaluation for produced or separated sperm form processing procedures was not evaluated yet.
Preparation for evaluation study to compare output of main used protocol Density Gradient (DG) with other protocol as swimming down using different study criteria can make more clear evaluation for better processing procedures.
The cases arranged for evaluation were semen samples are within WHO 2010 normal values for motility and count to be more than 5 million/ml. All samples to be divided to two portions for processing by the two preparation techniques.
Post preparation evaluation to be done for the two outputs using: Motility monitoring for 48 hours at 37.0 Dc, abnormal forms, hyaluronan affinity testing and contaminating bacterial growth.
In this study we will show drawback effect of physical and time stress interface applied on sperms during processing, and compare the out put results with that results produced from sperm processed without centrifugation interface or more processing time. As the sperm will separate it self by its natural movement and normal behavior.
Evaluation will be on three categories:
- The final output quality: as net highly progressive sperms and their count number ( sperm morphology, count and motility): as our results showed more phonologically normal sperm retrieved from SD method with higher count and more percentage of progressively motile sperms.
- The quality of sperm produced: through monitoring sperm vitality testing or sperm motility power among 2 days at 37.0 Ds. Motility will be evaluated after 24 hours and 48 hours. Both readings showed more decrease in sperm motility and vitality with time consuming with portions processed using density gradient. While portions processed using SD showed significant better readings. Another functional activity of sperm post processing is using HBA testing slide, as we evaluate the sperm affinity to hyaluronan. This Testing or assay indicate the physiological activity of sperm still maintained. While it showed HBA higher significant results with swimming down group study.
- Final evaluation, that are used to present the quality of separation: is monitored using the contamination and carry over effect during semen manipulation and processing. Bacterial contamination using routine culture media (Blood agar) to evaluate 5 microns from each portions for presence and absence of any bacterial cells or during processing and manipulation.
Study design
- Observational model
- Case-control
- Time perspective
- Prospective
- Number of arms
- 2
| Arm | Type | Receives |
|---|---|---|
| Density Gradient processing | – | Procedure: Diff-Quik staining for morphology evaluation, Procedure: Motility Evaluation for grade A and B, Procedure: Concentration (mil/ml), Procedure: 24 H half life, Procedure: 48 H half life, Procedure: HBA Testing, Biological: Bacterial contaminationSemen portion processed using Density Gradient (80/40 %) Spin on 1200 r.p.m./20min Wash on 1200 r.p.m./5min Rewash on 1200 r.p.m./5min then evaluation to be done using : Diff-Quik staining for morphology evaluation Motility Evaluation for grade A and B Concentration (mil/ml) 24 H half life 48 H half life HBA Testing Bacterial contamination |
| Swimming down Processing | – | Procedure: Diff-Quik staining for morphology evaluation, Procedure: Motility Evaluation for grade A and B, Procedure: Concentration (mil/ml), Procedure: 24 H half life, Procedure: 48 H half life, Procedure: HBA Testing, Biological: Bacterial contaminationSemen portion processed using swimming down (100 %) 30 min at room temperature processing for swim down Wash on 1200 r.p.m./5min Rewash on 1200 r.p.m./5min then evaluation to be done using : Diff-Quik staining for morphology evaluation Motility Evaluation for grade A and B Concentration (mil/ml) 24 H half life 48 H half life HBA Testing Bacterial contamination |
Who may qualify
- Ages
- 18 to 60 years
- Sex
- Male only
- Healthy volunteers
- Not accepted
- Age groups
- Adult
- Study population
- Cases arranged for study were submitted for routine semen analysis. Patient agreements collected as consent form signed during sample submission.
Key inclusion criteria (2 listed)
- Semen with motility more than 10 % according WHO 2010 standers
- Semen with sperm count more than 5 mil/ml according WHO 2010 standers
Key exclusion criteria (2 listed)
- Semen volume less than 1.0 ml
- Semen with liquefaction time more than 60 min
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 |
|---|---|---|
| Diff-Quik staining for morphology evaluationAlso: Abnormal forms %Papanicolaou stain (haematoxylin, orange G6, and EA50) gives a clear difference between basophilic and acidophilic cell constituents and thereby enables a detailed examination of chromatin pattern, which is useful in the evaluation of sperm morphology and assessment of presence of immature spermatozoa. Cytoplasmic staining can vary between red and green dependent on ionic strength, pH, and composition of the cell department and the stain (OG6 and EA50). Morphological assessment for processed portions of semen. As separation process has to minimize the morphological abnormal sperm. | Procedure or surgery | Density Gradient processing, Swimming down Processing |
| Motility Evaluation for grade A and BAlso: Motility %The better separation method will insure more progressive sperms after processing. Grading using A and B scale scored by (WHO 2010) references. | Procedure or surgery | Density Gradient processing, Swimming down Processing |
| Concentration (mil/ml)Also: Concentration for final outputcount of isolated sperms Using Makler chamber for count of sperm for each portion post processing. | Procedure or surgery | Density Gradient processing, Swimming down Processing |
| 24 H half lifeAlso: Motility after 24 hours incubation at 37 Dc.percentage of active motile to non motile sperms. Microscopic examination for each sample after 24 hours incubation, to evaluate the progressively motile sperm and those non progressive or non motile. | Procedure or surgery | Density Gradient processing, Swimming down Processing |
| 48 H half lifeAlso: Motility evaluation after 48hours incubation at 37 Dc.percentage of active motile to non motile sperms. Microscopic examination for each sample after 48 hours incubation, to evaluate the progressively motile sperm and those non progressive or non motile. | Procedure or surgery | Density Gradient processing, Swimming down Processing |
| HBA TestingAlso: Sperm physioogical activity evaluation HA bindingThe Hyaluronic Binding Assay (HBA®) is an important diagnostic tool for suspected male infertility in the analysis of semen. In a matter of minutes the HBA® slide provides an answer to the proportion of mature binding spermatozoa in the sample (The HBA® score %). By using HBA slide we can count the physiologically active sperms at each sample and calculate percentage of active sperms with reference to post processing count from 3rd investigating arm. | Procedure or surgery | Density Gradient processing, Swimming down Processing |
| Bacterial contaminationAlso: Bacterial contamination presence of post processinf samplesRoutine bacterial culture using Blood Agar culture plates for 10 micron finally washed samples, to confirm presence of any bacteria contamination caused by carry over during processing. | Biological | Density Gradient processing, Swimming down Processing |
Primary outcome
HBA
Time frame: 1 day
Affinity is measured according to percentage of non motile sperm produced from 10 µL of processed sample.
Abnormal forms
Time frame: 1 day
Percentage normally stained sperms after processing
3 secondary outcomes
Final concentration
Time frame: 1 day
24 Hour motility
Time frame: 1 day
48 Hour motility
Time frame: 2 days
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 1 Jan 2015.
- Background: PubMed 25466430Malvezzi H, Sharma R, Agarwal A, Abuzenadah AM, Abu-Elmagd M. Sperm quality after density gradient centrifugation with three commercially available media: a controlled trial. Reprod Biol Endocrinol. 2014 Dec 2;12:121. doi: 10.1186/1477-7827-12-121.
- Description: Density Gradient procedures descriptionLink provided by the sponsor.
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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.
Ahmad M Metwalley study director
Adliyah
2 sites in 2 countries: see every site and its status.
Questions about this trial
What is this clinical trial studying?
An observational study of Infertility, run by Ahmad Mustafa Mohamed Metwalley at 2 sites in 2 countries.
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 2 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 1 Jan 2015. Status can change between updates, so confirm with the study contact before making plans.
Where is the study taking place?
The record lists 2 sites in Bahrain, Egypt. 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?
HBA (time frame: 1 day). The study lists 2 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 NCT02329704, 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 1 Jan 2015.