An observational study in Infertility, sponsored by Ahmad Mustafa Mohamed Metwalley. Completed at 2 sites in 2 countries. Open to male participants aged 18 Years to 60 Years. Per ClinicalTrials.gov, last updated 2015-01-01.
Sponsored by Ahmad Mustafa Mohamed Metwalley · Observational
This study evaluates the quality of sperm produced after semen processing using Density Gradient (DG) and swimming down (SD).
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 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.
2,506 studies on the registry are indexed under Infertility; 407 are open to participants now.
This study's enrollment of 100 is below the median of 200 across 714 observational studies indexed under Infertility.
Browse Infertility studies →Ahmad Mustafa Mohamed Metwalley is the lead sponsor of 4 studies on the registry; none are open to participants now.
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Cases arranged for study were submitted for routine semen analysis. Patient agreements collected as consent form signed during sample submission.
Exclusion Criteria:
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
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
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
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
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.
Also known as: Abnormal forms %
The better separation method will insure more progressive sperms after processing. Grading using A and B scale scored by (WHO 2010) references.
Also known as: Motility %
count of isolated sperms Using Makler chamber for count of sperm for each portion post processing.
Also known as: Concentration for final output
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.
Also known as: Motility after 24 hours 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.
Also known as: Motility evaluation after 48hours incubation at 37 Dc.
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.
Also known as: Sperm physioogical activity evaluation HA binding
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.
Also known as: Bacterial contamination presence of post processinf samples
HBA
Affinity is measured according to percentage of non motile sperm produced from 10 µL of processed sample.
Time frame: 1 day
Abnormal forms
Percentage normally stained sperms after processing
Time frame: 1 day
Final concentration
Concentration was fluctuation according to original motility and concentration, DG
Time frame: 1 day
24 Hour motility
Motility evaluation after 37.0 Dc incubation
Time frame: 1 day
48 Hour motility
Motility evaluation after 37.0 Dc incubation
Time frame: 2 days
Bacterial contamination
bacterial culture for 5 micron of processed samples to insure absence of bacterial cells.
Time frame: 2 days
This study is completed, as verified in Dec 2014. You cannot join it, but the record below documents what was studied.
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Ahmad Mustafa Mohamed Metwalley