]>
IRCT20230528058312N3 IRCT 2024-02-19 Tahoora infertility treatment and lmited surgury clinic Exosome and stem cells therapy Investigating the effect of injecting the combination of exosomes extracted from follicular fluid, stem cells extracted from Wharton's jelly, platelets enriched from blood plasma and gonadotropins into the ovaries of poi, pof patients. 2024-02-20 anticipated 20 Complete https://irct.ir/trial/74927 interventional Randomization: N/A, Blinding: Not blinded, Placebo: Not used, Assignment: Parallel, Purpose: Treatment. 1 Premature Ovarian failure(POF) / Premature Ovarian Insufficiency (POI). Intervention 1: Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL).This group only autologous PRP(plate rich plasma) will be received. PRP was prepared from 40 mL of peripheral blood by the buffy coat protocol In brief, whole blood was divided into two aliquots and centrifugedat 3,000 rpm for 8 min at room temperature to form three layers.The top platelet poor plasma and the middle layerwere transferred to another two sterile tube and then centrifugedagain at 4,000 rpm for 5 min. After removal of upper portion of the supernatant, each pellet was reconstituted with 2.5 ml of supernatant. patient is lying in the dorsal lithotomy position, and a needle (Towako Trans myometrial Set 18 GA× 325 mm, Cook, Australia)with an angle of 15 to 30 degrees is advanced into ovarian stroma under vaginal ultrasound guidance (Honda 2000). Under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic will not be used, and the patient withstood the process without painful sensation. Intervention 2: Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL).this group receive exosome (100μg/ml)from the follicular fluid of fertile women referred to Tahura infertility center for IVF due to male factor infertility , who have been examined for HIV, HCV and HPV health and have grade A eggs, will be extracted according to the protocol provided by Eswaran with the help of Andreu Polyethylene glycol (PEG) (600 kDa). In order to check the presence of exosomes with the Mrestro brand nanodrop spectrophotometer, the concentration of DNA and RNA, with the ZEISS brand SEM electron microscope, the presence of exosomes will be checked. In addition, CD9, CD 63, CD81, CD82 factors will be checked with the help of Beckman brand flow cytometry.on the second and fifth day of the participants' menstrual cycle will be injected into their superficial layer of the ovary, only once during the treatment cycle.The patient is lying in the dorsal lithotomy position, and a needle (Towako Trans myometrial Set 18 GA× 325 mm, Cook, Australia)with an angle of 15 to 30 degrees is advanced into ovarian stroma under vaginal ultrasound guidance (Honda 2000). Under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic will be used, and the patient withstood the process without painful sensation. Intervention 3: Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL)only gonadotropin ,1 ml of 150 IU of FSH/75 IU of LH (Pergoveris,MerckSerono, Aubonne, Switzerland) was slowly infused into ovarian tissue, 0.5 ml in each ovary, under intravenous sedationusing 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood the process without painful sensation. No visible side effect will be noted in the whole procedure. Intervention 4: Intervention group: nfertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL).this group receive exosome (100μg/ml)and autologous PRP(4ml) , under intravenous sedationusing 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic will be used, and the patient withstood the process without painful sensation. Intervention 5: Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL)will be received gonadotropin 1(150IUrFSH/75 IU rLH, Pergoveris,MerckSerono, Aubonne, Switzerland) and exosome (100μg/ml)extracted from Follicular fluid of fertile woman injected under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood theprocess without painful sensation. No visible side effect is notedin the whole procedure. Intervention 6: Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL) receive gonadotropin 1ml(150IUrFSH/75IU rLH) (Pergoveris,MerckSerono, Aubonne, Switzerland) andautologous PRP (4mL) prepared by by the buffy coat protocol be injected under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood theprocess without painful sensation. Intervention 7: Intervention group: infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL)received gonadotropin 1ml(150IUrFSH/75IU rLH) (Pergoveris,MerckSerono, Aubonne, Switzerland) ,autologous PRP (4mL) prepared by by the buffy coat Porotocole and exosome (100μg/ml)extracted from Follicular fluid of fertile woman injected under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood theprocess without painful sensation. Intervention 8: Intervention group:Infertile women under the age of 40 for at least 4 months with primary or secondary amenorrhea with an increase in FSH serum level (above 25 mIU/mL) for this group, UC-MSC stem cells from human umbilical cord tissue, obtained from normal healthy deliveries, were donated for this group. They have come to be separated. Finally, Wharton jelly mesenchymal stem cells will be obtained through enzymatic digestion of umbilical cord cells using 1.67% collagenase (Sigma, C9891, Saint Louis, MO, USA) at 37°C.After dissociation, cells were lysed up to passage 5 using α-MEM (Gibco, 32561-102, Carlsbad, CA, USA) supplemented with 4 mM Glutamax (Gibco, 35050-079, Carlsbad, CA, USA) and bovine serum. Inactivated 10% will multiply. (Gibco, 16000044, Grand Island, NY, USA). Finally, vials containing pure UC-MSC mesenchymal stem cells using 6% hydroxyethyl starch (Claris, G/LVP-5, Ellisbridge, Ahmedabad, India) with 10% dimethyl sulfoxide (Sigma, D2650, Irvine, England). which will be frozen and quarantined by the vitrification method.Finally, randomly vials (before freezing and after thawing), in terms of life; Proliferation, mycoplasma, endotoxin, growth and differentiation will be reviewed. Before each treatment, cell vials will be selected according to the total number of viable cells obtained by quality controls (using flow cytometry and the Cell Counter device) after freezing and thawing (following the syringe preparation steps) to determine the required cell dose. need the protocol. Finally, to reach the therapeutic goal of 9 million stem cells in each infusion, vials containing stem cells will be adjusted. In addition, factors CD9, CD 63, CD81, CD82 will be evaluated with the help of Beckman Coulter brand flow cytometry. Finally, the stem cells extracted with the required concentration on day 2 and 5 will be added to the surface layer of the ovary if the patient is in dorsal lithotomy mode. Lying down with the help of a puncture needle (Towako (Transmyometrial Set 18 GA x 325 mm, Cook) with an angle of 15 to 30 degrees towards the ovary and with the guidance of the 2000 Honda vaginal ultrasound machine, it will be done only once during the treatment cycle.Before the injection, the patient will be put into artificial sleep with the help of an intravenous sedative using 100 mg of propofol (Fresenius Kabi Austria GmbH, Graz, Austria). No antibiotics will be used. According to previous studies, this process is painless and has no visible side effects. Yes - There is a plan to make this available What will be shared: All data obtained after identification of study cases can be shared. When: All information is accessible in April 2025. To whom: Our data will be available for all researchers and Doctors Conditions: All information related to the performed ultrasounds, tests related to exosome and stem cell extraction will be accessible Where to obtain: Applicants can contact one of the following email addresses to receive documents or data: asalighe@yahoo.com taba_1359@yahoo.com How to obtain: Three weeks after sending the applicant's message, the requested documents and files will be sent. Comments:
public Seyed Reza Tabatabaee Qomi
Tahoora infertility treatment and lmited surgury clinic,Tahoora street,Jamkaran
ghoum Iran (Islamic Republic of) 3747113135 +98 25 3725 4719 taba_1359@yahoo.com Tahoora infertility treatment and lmited surgury clinic scientific Seyed Reza Tabatabaee Qomi
Tahoora infertility treatment and lmited surgury clinic,Tahoora street,Jamkaran
Ghoum Iran (Islamic Republic of) 3747113135 +98 25 3725 4719 taba_1359@yahoo.com Tahoora infertility treatment and lmited surgury clinic Iran (Islamic Republic of) Women with premature menopause POF Women with premature ovarian failure POI Normal women with Grade A eggs 20 years 40 years Female Women with idiopathic infertility Women with adequate ovarian reserve N97.0 Female infertility associated with anovulation Treatment - Other Treatment - Other Treatment - Other Treatment - Other Treatment - Other Treatment - Other Treatment - Other Treatment - Other Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL).This group only autologous PRP(plate rich plasma) will be received. PRP was prepared from 40 mL of peripheral blood by the buffy coat protocol In brief, whole blood was divided into two aliquots and centrifugedat 3,000 rpm for 8 min at room temperature to form three layers.The top platelet poor plasma and the middle layerwere transferred to another two sterile tube and then centrifugedagain at 4,000 rpm for 5 min. After removal of upper portion of the supernatant, each pellet was reconstituted with 2.5 ml of supernatant. patient is lying in the dorsal lithotomy position, and a needle (Towako Trans myometrial Set 18 GA× 325 mm, Cook, Australia)with an angle of 15 to 30 degrees is advanced into ovarian stroma under vaginal ultrasound guidance (Honda 2000). Under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic will not be used, and the patient withstood the process without painful sensation. Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL).this group receive exosome (100μg/ml)from the follicular fluid of fertile women referred to Tahura infertility center for IVF due to male factor infertility , who have been examined for HIV, HCV and HPV health and have grade A eggs, will be extracted according to the protocol provided by Eswaran with the help of Andreu Polyethylene glycol (PEG) (600 kDa). In order to check the presence of exosomes with the Mrestro brand nanodrop spectrophotometer, the concentration of DNA and RNA, with the ZEISS brand SEM electron microscope, the presence of exosomes will be checked. In addition, CD9, CD 63, CD81, CD82 factors will be checked with the help of Beckman brand flow cytometry.on the second and fifth day of the participants' menstrual cycle will be injected into their superficial layer of the ovary, only once during the treatment cycle.The patient is lying in the dorsal lithotomy position, and a needle (Towako Trans myometrial Set 18 GA× 325 mm, Cook, Australia)with an angle of 15 to 30 degrees is advanced into ovarian stroma under vaginal ultrasound guidance (Honda 2000). Under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic will be used, and the patient withstood the process without painful sensation. Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL)only gonadotropin ,1 ml of 150 IU of FSH/75 IU of LH (Pergoveris,MerckSerono, Aubonne, Switzerland) was slowly infused into ovarian tissue, 0.5 ml in each ovary, under intravenous sedationusing 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood the process without painful sensation. No visible side effect will be noted in the whole procedure. Intervention group: nfertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL).this group receive exosome (100μg/ml)and autologous PRP(4ml) , under intravenous sedationusing 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic will be used, and the patient withstood the process without painful sensation. Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL)will be received gonadotropin 1(150IUrFSH/75 IU rLH, Pergoveris,MerckSerono, Aubonne, Switzerland) and exosome (100μg/ml)extracted from Follicular fluid of fertile woman injected under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood theprocess without painful sensation. No visible side effect is notedin the whole procedure. Intervention group: Infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL) receive gonadotropin 1ml(150IUrFSH/75IU rLH) (Pergoveris,MerckSerono, Aubonne, Switzerland) andautologous PRP (4mL) prepared by by the buffy coat protocol be injected under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood theprocess without painful sensation. Intervention group: infertile women with primary or secondary amenorrhea for more than 4 months under 40 years old with an elevated serum level of FSH (higher than 25 mIU/mL)received gonadotropin 1ml(150IUrFSH/75IU rLH) (Pergoveris,MerckSerono, Aubonne, Switzerland) ,autologous PRP (4mL) prepared by by the buffy coat Porotocole and exosome (100μg/ml)extracted from Follicular fluid of fertile woman injected under intravenous sedation using 100 mg propofol (Fresenius Kabi Austria GmbH, Graz,Austria). No antibiotic is used, and the patient withstood theprocess without painful sensation. Intervention group:Infertile women under the age of 40 for at least 4 months with primary or secondary amenorrhea with an increase in FSH serum level (above 25 mIU/mL) for this group, UC-MSC stem cells from human umbilical cord tissue, obtained from normal healthy deliveries, were donated for this group. They have come to be separated. Finally, Wharton jelly mesenchymal stem cells will be obtained through enzymatic digestion of umbilical cord cells using 1.67% collagenase (Sigma, C9891, Saint Louis, MO, USA) at 37°C.After dissociation, cells were lysed up to passage 5 using α-MEM (Gibco, 32561-102, Carlsbad, CA, USA) supplemented with 4 mM Glutamax (Gibco, 35050-079, Carlsbad, CA, USA) and bovine serum. Inactivated 10% will multiply. (Gibco, 16000044, Grand Island, NY, USA). Finally, vials containing pure UC-MSC mesenchymal stem cells using 6% hydroxyethyl starch (Claris, G/LVP-5, Ellisbridge, Ahmedabad, India) with 10% dimethyl sulfoxide (Sigma, D2650, Irvine, England). which will be frozen and quarantined by the vitrification method.Finally, randomly vials (before freezing and after thawing), in terms of life; Proliferation, mycoplasma, endotoxin, growth and differentiation will be reviewed. Before each treatment, cell vials will be selected according to the total number of viable cells obtained by quality controls (using flow cytometry and the Cell Counter device) after freezing and thawing (following the syringe preparation steps) to determine the required cell dose. need the protocol. Finally, to reach the therapeutic goal of 9 million stem cells in each infusion, vials containing stem cells will be adjusted. In addition, factors CD9, CD 63, CD81, CD82 will be evaluated with the help of Beckman Coulter brand flow cytometry. Finally, the stem cells extracted with the required concentration on day 2 and 5 will be added to the surface layer of the ovary if the patient is in dorsal lithotomy mode. Lying down with the help of a puncture needle (Towako (Transmyometrial Set 18 GA x 325 mm, Cook) with an angle of 15 to 30 degrees towards the ovary and with the guidance of the 2000 Honda vaginal ultrasound machine, it will be done only once during the treatment cycle.Before the injection, the patient will be put into artificial sleep with the help of an intravenous sedative using 100 mg of propofol (Fresenius Kabi Austria GmbH, Graz, Austria). No antibiotics will be used. According to previous studies, this process is painless and has no visible side effects. Absence of eggs in the ovaries of women under 40 years and primary or secondary amenorrhea for more than 4 months in women under 40 years with an elevated serum level of Follicular Stimulated Hormone(higher than 25 mIU/mL. Timepoint: 15 days after the injection and then every month at the time of ovulation during one year. Method of measurement: Transvaginal ultrasound device, Hormonal Test by Elisa Kit. Absence of eggs in the ovaries of women under 40 years and primary or secondary amenorrhea for more than 4 months in women under 40 years with an elevated serum level of Follicular Stimulated Hormone(higher than 25 mIU/m. Timepoint: 15 days after the injection and then every month at the time of ovulation during one year. Method of measurement: Transvaginal ultrasound device, Hormonal Test by Elisa Kit. Tahoora infertility treatment and lmited surgury clinic Approved 2024-02-19 Islamic Azad University, Qom branch 15 Khordad St., Islamic Azad University, next to Social Security Insurance, Qom branch, Iran Ghoum Ghoum Iran (Islamic Republic of)