THE INFLUENCE OF EJACULATORY ABSTINENCE INTERVALS ON SEMEN QUALITY - OLD CONCEPT, NEW EVIDENCES

Transcription

1 THE INFLUENCE OF EJACULATORY ABSTINENCE INTERVALS ON SEMEN QUALITY - OLD CONCEPT, NEW EVIDENCES Edson Borges Jr 1,2, Amanda Setti Raize 1,2 ; Daniela Paes de Almeida Ferreira Braga 1,2 Rodrigo R. Provenza 1 ; Rita de Cássia, Sávio Figueira 1, Assumpto Iaconelli Jr. 1

2 INTRODUCTION It is estimated that nearly 72.4 million couples globally experience fertility problems Infertility: 10% of the couples World Health Organization (WHO) Male factor Others Sperm count Sperm motility Sperm morphology Normal Abnormal

3 INTRODUCTION Ejaculatory abstinence (EA) ejaculatory abstinence Conventional semen analysis 2 7 days WHO 3 4 days ESHRE semen quality?

4 INTRODUCTION Semen volume DNA fragmentation Sperm concentration EA \\\\\\\\ Chromatin quality Sperm motility Sperm viability (De Jonge et al., 2004, Gosalvez et al., 2011, Sanchez-Martin et al., 2013, Agarwal et al., 2016).

5 INTRODUCTION EA Pregnancy Ejaculatory Abstinence ICSI outcomes (Jurema et al., 2005, Marshburn et al., 2010, Sanchez-Martin et al., 2013). Scarce studies Limited number of subjects

6 OBJECTIVE To investigate the influence of ejaculatory abstinence on basic and advanced semen parameters To investigate the influence of ejaculatory abstinence on the outcomes of ICSI

7 MATERIALS AND METHODS STUDY DESIGN Prospective cohort study Influence of EA intervals on semen quality Patients undergoing conventional semen analysis 818 pacientes Couples undergoing ICSI 483 couples Influence of EA intervals on ICSI outcomes.

8 MATERIALS AND METHODS EVALUATED VARIABLES: FIRST ANALYSIS Semen volume Sperm count Sperm motility Total motile sperm count Morphology MSOME Sperm DNA fragmentation EVALUATED VARIABLES: SECOND ANALYSIS Fertilisation rate High-quality embryos rate on day 3 Blastocyst formation rate on day 5 Number of transferred embryos Implantation rate Pregnancy rate Miscarriage rate

9 MATERIALS AND METHODS Ovarian stimulation rfsh GnRH antagonist Semen analysis Patients filled a form regarding the EA interval Semen samples collected in the lab by masturbation Threshold values (WHO 2010) Semen preparation 2-layered density gradient centrifugation ICSI Performed according to Palermo et al. 1992

10 MATERIALS AND METHODS Abnormal shape MOTILE SPERM ORGANELLE MORPHOLOGY EXAMINATION Small vacuoles Large vacuoles Aliquot of sperm cell transferred to a microdroplet of 8% PVP in a glass dish 200 sperm cells per sample analyzed under x6,600 Presence and size of vacuoles were recorded

11 MATERIALS AND METHODS MSOME: SPERM CLASSIFICATION (Vanderzwalmen et al., 2008) Grade I: Normal form No vacuoles Grade II: Normal form 2 small vacuoles Grade III: Normal form > 2 small vacuoles or at least one large vacuole Grade IV: Abnormal head shapes or other abnormalities Large vacuole

12 MATERIALS AND METHODS SPERM DNA FRAGMENTATION ANALYSIS Denaturation Lysis Wash 20 x 10 6 /ml agarose 90 º C Halosperm G2, Halotech Fixation Staining sperm chromatin dispersion test

13 MATERIALS AND METHODS EMBRYO MORPHOLOGY AND EMBRYO TRANSFER

14 RESULTS Linear model analysis of the association between sperm parameters and EA length (n = 818) SEMEN PARAMETER R SLOPE R 2 (%) P-VALUE Semen volume (ml) <0.001 Sperm count (x10 6 /ml) <0.001 Total sperm count (x10 6 ) <0.001 Total sperm motility (%) Progressive sperm motility (%) TMSC (x10 6 ) <0.001 Morphology (%) MSOME grade I (%) MSOME grade II (%) MSOME grade III (%) MSOME grade IV (%) SDF (%) <0.001

15 RESULTS Linear model analysis of the association between sperm parameters and EA length (n = 818) SEMEN PARAMETER R SLOPE R 2 (%) P-VALUE Semen volume (ml) <0.001 Sperm count (x10 6 /ml) <0.001 Total sperm count (x10 6 ) <0.001 Total sperm motility (%) Progressive sperm motility (%) TMSC (x10 6 ) <0.001 Morphology (%) *TMSC: total motile sperm count

16 RESULTS Linear model analysis of the association between sperm parameters and EA length (n = 818) SEMEN PARAMETER R SLOPE R 2 (%) P-VALUE MSOME grade I (%) MSOME grade II (%) MSOME grade III (%) MSOME grade IV (%) SDF (%) <0.001 *MSOME: motile sperm organelle morphology *SDF: Sperm DNA fragmentation

17 RESULTS Linear model analysis of the association between ICSI outcomes and EA length (n = 483) VARIABLE R SLOPE R 2 P-VALUE Fertilisation rate High-quality embryos rate on day Blastocyst formation rate on day Number of transferred embryos Implantation rate VARIABLE OR 95% CI P-VALUE Pregnancy rate Miscarriage rate

18 RESULTS Linear model analysis of the association between ICSI outcomes and EA length (n = 483) VARIABLE R SLOPE R 2 P-VALUE Fertilisation rate High-quality embryos rate on day Blastocyst formation rate on day Number of transferred embryos Implantation rate VARIABLE OR 95% CI P-VALUE Pregnancy rate Miscarriage rate

19 DISCUSSION Sperm volume Sperm count TMSC SDF LONGER EA INTERVALS Fertilization rate Blastocyst formation rate

20 DISCUSSION Longer EA periods Long periods of EA Senescence of spermatozoa Sperm acrosin activity Sperm-egg interaction, Sperm zona pellucida binding Egg penetration (Blackwell & Zaneveld 1992)

21 DISCUSSION EA EA Graded I-IV by MSOME EA Sperm DNA Fragmentation

22 DISCUSSION Kourouma et al., 2015

23 DISCUSSION Sperm membrane damage through lipid peroxidation Excessive ROS Oxidative stress Sperm protein damage Sperm DNA susceptibility to free radical attack

24 DISCUSSION Fertilization rate EA Blastocyst formation Pregnancy outcomes Oxidative damage Fertilization rate

25 DISCUSSION ICSI bypasses natural sperm selection Injected sperm DNA fragmentation Early and late paternal effects Impaired fertilization and blastocyst formation

26 DISCUSSION WHO recommendation of EA for assessing conventional semen parameters (2-7 days) EA based on copulatory regularity Couples trying to conceive naturally are encouraged to have intercourse every other day during the fertile period Detrimental effects on sperm function Does not endanger sperm quality

27 CONCLUSION Prolonged EA periods can have a detrimental effect on both sperm function and quality Shortening the EA intervals could optimize sperm quality, fertilization and blastocyst formation Documentation that shortening the EA period would lessen these potentially harmful influences will require further study

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