Endocrine Disrupting Chemicals: Potential Effects on Male and Female Reproductive Health in Saskatchewan
Condition: Unexplained Infertility; Polycystic Ovarian Syndrome; Male Infertility
MeSH terms: Polycystic Ovary Syndrome; Infertility; Infertility, Male
Study type: Observational [Patient Registry]

Summary: 
      The overall objective of this study is to determine whether serum BPA and/or phthalate
      concentrations differ in fertile versus infertile men and women in Saskatchewan. The
      investigators will test the following hypothesis:

        1. Serum BPA and/or phthalate concentrations will be greater in women with unexplained
           infertility or PCOS compared to a control group

        2. Serum BPA and/or phthalate concentrations will be greater in men with male factor
           infertility compared to a control group
    

Detailed description: 
      Study Sample:

      Couples undergoing in vitro fertilization (IVF) and/or Intracytoplasmic Sperm Injection
      (ICSI) for assisted reproduction at ARTUS as well as healthy male and female volunteers 18-43
      years of age will be asked to participate. In order for infertility patients to be eligible,
      both the male and female partners must agree to participate. Couples with a combination of
      both male and female factor infertility will not be eligible to participate. Couples
      undergoing infertility treatment for the following reasons will be evaluated:

        -  unexplained infertility (n=15 men, n=15 women)

        -  PCOS (n=15 men, n=15 women)

        -  Male factor infertility (n=15 men, n=15 women)

      Couples with male factor infertility will serve as controls for the female comparisons (i.e.,
      follicular fluid analyses), and couples with PCOS will serve as controls for the male
      comparisons (i.e., seminal plasma analyses).

      Additional participants will be assigned to the following control groups (for blood sample
      analyses):

      -Healthy volunteers with no history or current diagnosis of reproductive dysfunction (n=15
      men, n=15 women)

      Participant Recruitment:

      Infertile couples will be recruited from the clinical practices of Drs. Allison Case and
      Adrian Gamelin at the Assisted Reproductive Technologies Centre at the University of
      Saskatchewan (ARTUS). Posters for recruiting ARTUS patients will also be placed in the
      waiting room and examining rooms in the Dept of OB/GYN, Division of REI. Healthy male and
      female volunteers will be recruited from advertisements on the PAWS website at the University
      of Saskatchewan and Health E-News newsletters in the Saskatoon Health Region.

      Pre-Study Visit:

      Eligible participants will be asked to visit the ARTUS Centre for a pre-study appointment. At
      this appointment, study procedures will be discussed with participants and consent will be
      obtained.

      Study Visits:

      Blood and follicular fluid samples will be obtained from female partners of infertile
      couples. Blood and seminal plasma samples will be obtained from male partners of infertile
      couples. Only blood samples will be collected from healthy men and women. All samples will be
      collected in both fertile and infertile couples over a 2 week time interval.

      A. Infertile couples:

      Infertile couples participating in this study will undergo fertility treatment as per the
      normal standard of care. In other words, the fertility medications, ultrasound assessments,
      blood work during treatment, oocyte retrieval, semen collection, laboratory procedures and
      embryo transfer procedures will not be affected by participating in this study.

      Both the male and female partner will be asked to provide 1 extra blood sample (approximately
      7 mL or 1 teaspoon of blood) on each of the following regularly-scheduled appointments:

        1. First ultrasound visit (approximately day 1 of FSH therapy)

        2. Day of egg retrieval

        3. Day of embryo transfer

      Women normally undergo regular blood draws during their IVF treatment; therefore, one
      additional tube will be drawn for research purposes at the regularly-scheduled blood draw
      appointments. Men do not normally provide a blood sample during IVF treatment, but will be
      asked to do so in this study. Blood draws will be performed by phlebotomists (staff that
      specialize in drawing blood) at Royal University Hospital, physicians, nurses and/or
      researchers with training in phlebotomy. The blood samples will be processed to remove the
      cells, and the serum will be frozen for 12-24 months until the completion of the study.

      The follicles in the woman's ovaries will be drained and the eggs will be retrieved from the
      follicular fluid, as per the standard of care. After the eggs are collected, the follicular
      fluid is normally thrown away. In this study, we will collect the fluid from the follicles on
      each ovary after the eggs are retrieved. The follicular fluid will be frozen for 12-24 months
      until study completion.

      A semen sample will be provided by the man on the day of the oocyte retrieval procedure, as
      per standard of care. The semen sample is processed by allowing the semen to pass through a
      column to separate the most motile sperm (moving very quickly) from the less motile and
      non-motile sperm. The seminal plasma is the fluid portion of the semen that remains after the
      most motile sperm have been obtained. The seminal plasma is normally thrown away after the
      motile sperm are obtained. In this study, we will collect the seminal plasma and freeze it
      for 12-24 months until the study is completed.

      B. Healthy Volunteers:

      Healthy volunteers will be asked to provide 1 blood sample (approximately 7 mL or 1 teaspoon
      of blood) on 3 separate days. Blood draws will be performed by physicians, nurses and/or
      researchers with training in phlebotomy (i.e., drawing blood). Appointments for blood draws
      will be made approximately 4-7 days apart.

      At the completion of the study all of the frozen blood, follicular fluid, and seminal plasma
      samples will be transported to the Toxicology Center at the University of Saskatchewan. The
      levels of exposure to BPA and phthalates will be evaluated. The samples will be destroyed
      upon completion of the study. The expected duration of study participation is approximately
      13-20 days (in accordance with standard fertility therapy) for fertility patients and 3 days
      for healthy volunteers. All samples will be obtained over a 2 week time period.

      Statistical Analysis

      Include a summary of the primary and secondary end-points/outcomes, the planned sample size
      (with justification) and planned statistical and interim analyses.

      Study Endpoints:

      Exposure to phthalates will be determined by quantifying concentrations of 5 different
      pthalates. Concentrations of Bisphenol A (BPA), di-2-ethylhexyl phthalate (DEHP), di-n-octyl
      phthalate (DNOP), di-n-butyl phthalate (DNBP), n-butyl benzyl phthalate (BBP), diethyl
      phthalate (DEP), diisononyl phthalate (DINP) and diisodecyl phthalate (DIDP) will be
      quantified in the blood cells, blood plasma, and follicular fluid (individual and pooled
      samples) by use of liquid chromatography and tandem mass spectrometry methods already
      developed in our laboratory. Analyses will be conducted by use of an Agilent 1200 series High
      Performance Liquid Chromatography (HPLC) system interfaced to an ABI SCIEX 3200 triple
      quadrapole mass spectrometer system or to a QToF II quadrapole time of flight mass
      spectrometer (Micromass, Beverly MD). Toxicant concentrations will be the dependent
      variables. Reproductive status (i.e, unexplained infertility, PCOS, male factor infertility,
      controls) will be the independent variable. Concentrations of BPA and phthalates in blood
      cells, blood plasma, seminal plasma and pooled follicular fluid will be compared between
      study groups using multivariate Analyses of Variance (ANOVA).

      n=15 per group is appropriate for conducting this pilot study.

      Potential Significance/Justification:

      Greater knowledge about the possible effects of BPA and phthalates on reproductive function
      may increase our understanding of the causes of infertility, assist in preventing
      reproductive disease, and optimize reproductive health, which will thereby improve patient
      care. We believe that the results of our study, whether positive or negative, will be of
      interest to the general public, health professionals and scientific community.
    

Observational model: Cohort
Time perspective: Prospective
Sampling method: Probability Sample
Study population: 
        Infertile couples with unexplained infertility, male factor infertility and PCOS Healthy
        volunteers with history of fertility
      
Biospecimen description: 
      Blood, Follicular Fluid and Seminal Plasma Samples
    
Keywords: Bisphenol A; phthalates; infertility; human