The Effect of Astaxanthin Administration on Oxidative Stress Markers and Inflammatory Cytokines in the Follicular Fluid of Patients with Poor Ovarian Response: A Triple-Blind, Randomized control trial

Therefore, the aim of this study is to investigate the effect of astaxanthin on assisted reproductive technology results and the oxidative stress and inflammatory cytokines markers and cell free DNA in patients with poor ovarian response.

Two arm parallel group, Triple-Blind, Randomized control trial and outcome assessment Randomization will be in the form of blocking. 60patients with poor ovarian response will enter the project. Patients will be randomly divided into two groups: group one will receive 12 mg of astaxanthin (2 capsules) per day and group two will receive placebo capsules for 8 weeks.

The location of the experiment: Tehran University of Medical Sciences. Sampling location: Omid fertility center. This clinical trial is a randomized, triple-blind placebo-controlled trial. In any clinical trial, three groups can influence the results of the study, these three groups are 1- the participants in the research, 2- the implementers of the plan, 3- the evaluators or others who analyze the obtained data. If blinding is done for all three groups involved, it is called a triple-blind study.

The inclusion criteria were: age > 35 years, and poor ovarian reserve (anti-müllerian hormone (AMH) < 1.2 ng/ml, or antral follicle count (AFC) < 5) (indicating low prognosis group 4 following the POSEIDON stratification) (24). Exclusion criteria: including history of ovarian surgery or chemotherapy, autoimmune or endocrine disease, severe male factor infertility

Astaxanthin consumer. Placebo consumer.

1- ART outcomes: Evaluation of the number of MII oocytes & retrieved oocytes, number & quality of embryos, fertilization rate, chemical & clinical pregnancy rate 2- Evaluation of MDA, SOD, TAC, IL-6, IL-8, VEGF in serum and follicular fluid, & Cell-free DNA levels in follicular fluid.

The precise age range under study was determined to ensure the homogeneity of the patient population with poor ovarian response. The selected age range was 36-41 years, corresponding to group 4 of the POSEIDON classification. Additionally, modifying certain variables such as adding variables that include the cycle cancellation rate and the dosage of ovarian stimulation medications which is important in patients with poor ovarian response.

Eligible patients will be randomly assigned to the study after visiting the gynecology clinic. Randomization will be in the form of blocking in such a way that the method of replacement blocks is used within each floor. The blocks will be binary, consisting of AB and BA. The randomization tool will be a table of random numbers, that using the sequence AB for numbers zero through 4 in the random number table and the sequence BA for 5 to 9.It should be noted that the letter A means the intervention group and the letter B means the control group For hiding we use concealment allocation in a way that before the individual is assigned, the assigned group is not specified. In this method, each of the random sequences created on a cards are placed in the closed envelopes. In order to maintain a random sequence, the envelopes are numbered in the same way on the outer surface. Finally, the lids of the envelopes are glued and placed inside a box, respectively. At the time of registration of participants, based on the order of entry of the eligible participants in the study, one of the closed envelopes will be opened and the assigned group of the participant will be revealed.

This study will be a three-way blind and the patient, researcher and statistical analyst of the patients' group therapy have no information and the decoder is outside the research team. The drug under study (Astaxanthin) and placebo in a completely similar shape and appearance, each of which is in the same bottle instead of the drug will be provided to the subjects based on random allocation. It should be noted that the drug content of each The bottles are code-tagged on the bottles by the by someone outside the research team consultant and the research team is not aware of their interpretation.

All data obtained in the research will be published in one of the valid and related scientific journals.

2025

Researchers working in academic institutions

The data of this study can be used as an adjuvant therapy in patients with poor ovarian response underwent IVF/ICSI.

Anahid Shafie

By sending an email to the corresponding author.