Clinical trial NCT04009473: Stem Cell Therapy and Growth Factor Ovarian in Vitro Activation
Official title: Platelet Rich Plasma Ovarian in Vitro Activation and Stem Cells Transplantation In Women With Ovarian Failure
Study type: INTERVENTIONAL
Overall status: UNKNOWN
Phase: PHASE1
PHASE2
Conditions: Menopause
Menopause Ovarian Failure
Ovarian Failure
Premature Ovarian Failure
Premature Ovarian Failure 1
Premature Ovarian Failure 2A
Premature Ovarian Failure 2B
Premature Ovarian Failure 3
Premature Ovarian Failure 4
Premature Ovarian Failure 5
Premature Ovarian Failure 6
Premature Ovarian Failure 7
Premature Ovarian Failure 8
Premature Ovarian Failure 9
Premature Ovarian Failure 10
Premature Ovarian Failure 11
Premature Ovarian Failure 12
Premature Ovarian Failure 13
Premature Ovarian Failure 14
Ovarian Insufficiency
Ovarian Failure Secondary
Ovarian Failure, Premature
Ovarian Insufficiency, Primary
Menopause, Premature
Keywords: platelet rich plasma
growth factors
stem cells
bone marrow
in vitro activation
ovarian cortex transplantation
hippo signalling pathway
Lead sponsor: Forever Young d.o.o.
Sponsor class: OTHER
Collaborators: name: Medigroup Health System, Jevremova hospital
name: Saint James Hospital Malta
name: Re-medika Hospital Skoplje
First posted: 2019-07-05
Enrollment: 100
Sex: FEMALE
Minimum age (years): 25
Standard ages: ADULT
OLDER_ADULT
Accepts healthy volunteers: no
Allocation: NA
Intervention model: SINGLE_GROUP
Intervention model description: Pre-post study design:

A study group receives the intervention, and the changes in parameters were measured and compared before and after the intervention.
Primary purpose: TREATMENT

Brief summary: SEGOVA procedure includes - Stem cell therapy, G - Growth factor Platelet Plasma Rich therapy and in Vitro Activation of the ovaries.

Detailed description: The current study seeks to demonstrate how integrative ovarian rejuvenation program called SEGOVA influence the restoration of hormone stability and increase in the number of follicles in ovarian failure patients. The recruitment of patients and data collection would be performed at three sites: The Special Hospital Jevremova Belgrade, Saint James Hospital Malta, and Remedica Skopje Hospital, Macedonia. A 50-100 of Infertile women with ovarian failure will be included in the period between July 2019 - December 2021. The patients would be coded when entering the program, and all personal information would be protected. Information about medical treatment and background will be held on both paper and electronic case report forms.The result of the procedure will be maintained in the research database with blinding to clinical physician that will perform post procedure follow-up of those patients. In case of a complication, possibly related to treatment, the case will be unblinded.

The first day of procedure, autologous platelet rich plasma (PRP) is obtained from 100-120 mL of whole blood. After separation 4 mL of PRP is obtained. Concentration of platelets is optimised on 6-8x baseline and 2-3x leucocytes baseline. For activation of PRP autologous thrombin is used in ratio 1:10.

On the same day the laparoscopic cortical resection of the ovary is performed in standard technique. Ovarian cortex sample is minced in a petri dish.The volume of activated PRP (4 mL) is mixed with ovarian fragments and incubated for the next 48h at 37℃ and 5% CO2. After 48h, the bone marrow sampling from tibia under general anesthesia is performed to obtain bone marrow. After centrifugation the Bone Marrow Aspirate Concentrate (BMAC) with nucleated cells is obtained (3 +/- 1,5 mL).

Finally, after the 48h has passed from ovarian cortex tissue incubation, fragmented tissue of the ovary with 4 mL of PRP will be injected together with 3 +/- 1.5 mL BMAC into the subcortical region of the both ovaries as transvaginal ultra sound guided injection.

After the procedure, during one year follow up, hormone levels of follicle stimulating hormone (FSH), luteinizing hormone (LH),estradiol (E2), progesterone (PG) and anti-mullerian hormone (AMH) would be measured and the follicle numbers would be monitored to provide insight regarding the ovarian function.To investigate the changes in hormone levels, comparisons of the levels detected prior to the intervention, and 3, 6 and 12 months post-intervention, would be performed using Wilcoxon's rank test. Descriptive statistics will be provided for each variable as mean or median number, frequency, percentage, table or graph.

In patients with oocytes retrieved after SEGOVA procedure, standard In Vitro Fertilization protocol would be performed,and fertilization, cleavage and clinical pregnancy rates will be monitored.

Eligibility criteria: Inclusion Criteria:

Confirmed informed consent, signature and date

* A woman over 25 years of age
* Primary or secondary amenorrhea for at least 3 months
* Hormone Anti Mullerian Hormone values \<_0.42 ng / ML and Follicle stimulating hormone FSH\> 20 IU / L, and / or failure of previous attempts of assisted reproductive techniques due to limited ovarian response (less than 3 oocyte cells obtained).
* The proper karyotype 46, XX.
* Presence of at least one ovary

Exclusion Criteria:

* Currently pregnancy or breastfeeding
* Presence of Sexually Transmitted Disease (STD positive)
* There is presence of acute infection (C Reactive Protein\>5)
* There is an anamnesis or evidence of existing gynecological malignancy
* The presence of adnexal masses indicating the need for further evaluation.
* It has a contraindication to laparoscopic surgery and / or general anesthesia
* Over the past two weeks, use of the following medicines: Oral or systemic corticosteroids, hormones (estrogen, progestins, oral contraceptives), Danazol, anticoagulants, herbal or botanical supplements with potential hormonal effects.
* Type I diabetes mellitus
* Known significant anemia (hemoglobin \<8 g / dL), Severe venous thrombosis and / or pulmonary embolism, Cerebrovascular disease, Presence of heart disease, Premature kidney disease (defined as urea in the blood\> 30 mg / dL or serum creatinine\> 1.6 mg / dL).

Primary outcomes:
measure: Oocyte presence
timeFrame: 12 months
measure: Changes in Hormone Levels of Follicle-stimulating hormone FSH (mIU/mL)
timeFrame: 12 months
measure: Changes in Hormone Levels of Luteinizing Hormone (mIU/mL)
timeFrame: 12
measure: Changes in Hormone Levels of Estradiol E2 (pg/mL)
timeFrame: 12
measure: Changes in Hormone Levels of Progesterone PG (ng/mL)
timeFrame: 6
Secondary outcomes:
measure: Fertilization and implantation rate
timeFrame: 12 months
measure: Number of participants with Positive Clinical Pregnancy
timeFrame: 24 months
measure: Changes in total score of The World Health Organization Quality of Life (WHOQOL) BREF modified scale
timeFrame: 12 months
measure: Changes in anti-Mullerian hormone (AMH) levels (ng/mL)
timeFrame: 6 months
Other outcomes:
Locations:
city: Sliema
country: Malta
facility: Saint James Hospital Malta
city: Skopje
country: North Macedonia
facility: Re-medika Hospital
state: Macedonia
city: Belgrade
country: Serbia
facility: Medigroup Health System, Jevremova hospital
Study officials:
affiliation: Medigroup, Belgrade, Serbia
name: Aleksandar Ljubic, MD, PhD
role: PRINCIPAL_INVESTIGATOR
References:
citation: Tinjic S, Abazovic D, Ljubic D, Vojvodic D, Bozanovic T, Ibrisimovic M, Markovic S, Ljubic A. Influence of Autologous In Vitro Activation of Ovaries by Stem Cells and Growth Factors on Endocrine and Reproductive Function of Patients with Ovarian Insufficiency-A Clinical Trial Study. Int J Fertil Steril. 2021 Jul;15(3):178-188. doi: 10.22074/IJFS.2020.134678. Epub 2021 Jun 22.