Ozone sauna therapy and pulsed electromagnetic field therapy could potentially improve outcome in women with diminished ovarian reserve undergoing assisted reproductive technology

Abstract

There are limited treatment options for women with severely diminished ovarian reserve (DOR) who experience repeatedly failed in vitro fertilization (IVF) cycles and with persistently thin endometrial lining thickness (EMT) during frozen embryo transfer cycles. Therefore, a large majority of patients resort to using donor oocytes and gestational carriers. Data from existing animal and human studies suggest that ozone sauna therapy (OST) and pulsed electromagnetic field therapy (PEMF) are emerging as potential therapeutic adjuncts for female reproduction. This study was conducted to assess the fertility outcome of OST + PEMF in vivo in patients undergoing IVF/frozen embryo transfe and the effects of OST in vitro on human granulosa cell (GC) function. Forty-four women with DOR underwent their 1^st IVF cycle (Cycle 1), and then were administered transdermal and intravaginal OST + PEMF, twice a week for 3 weeks, followed by a 2^nd IVF cycle (Cycle 2) using the same protocol as in Cycle 1. GCs collected from another six women who underwent egg retrieval were equally split and cultured with OST (test) or placed in room temperature (control) outside the OST chamber in the same room. The results demonstrated that Cycles 1 and 2 had no significant difference in the number of days of stimulation, baseline hormones measured, number of oocytes retrieved or peak estradiol levels. However, the number of embryos formed after OST + PEMF in Cycle 2 was significantly higher than the Cycle 1. Furthermore, EMT measured in Cycle 2 demonstrated a significant increase compared to Cycle 1 and all patients reached a satisfactory EMT of approximately 7 mm. In vitro studies demonstrated that OST led to a 5-fold significant increase in the aromatase enzyme while a significant 50% reduction was noted in the side-chain cleavage enzyme in GCs. Both OST + PEMF are known for their vasodilatory, anti-inflammatory, and antioxidant actions, which could enhance endometrial receptivity and increase the number of formed embryos without increasing the number of oocytes retrieved, suggesting an improvement in oocyte quality. Finally, ozone can alter genes involved in steroidogenesis suggesting that it could improve ovarian function.

Keywords: diminished ovarian reserve; endometrial lining thickness; fertilization; granulosa cells; in vitro; oocyte; ozone sauna therapy; pulsed electromagnetic field therapy; steroidogenesis.

Figure 1:

Ozone therapy and pulsed electromagnetic field therapy improve the female reproductive system. Note: Women (n = 44) with severe diminished ovarian reserve underwent 1^st in vitro fertilization cycle (Cycle 1), followed by hyperthermic ozone & carbonic acid transdermal therapy machine use for 3 weeks followed by a 2^nd in vitro fertilization cycle (Cycle 2) using the same protocol as Cycle 1. There was no significant difference in the number of oocytes collected or peak serum estradiol (E2) levels. However, the endometrial lining thickness (EMT) and number of embryos formed were significantly higher in Cycle 2 compared to Cycle 1. Data are presented as mean ± SEM and were analyzed by paired t-test. The detailed statistical analysis is shown in Additional Table 1.

Figure 2:

Ozone therapy and pulsed electromagnetic field therapy improve the genes involved in steroidogenesis of primary human granulosa cells. Note: Six participants underwent oocyte retrieval had their granulosa cells split equally and cultured, then half of the cells placed in the hyperthermic ozone & carbonic acid transdermal therapy (HOCATT) machine (ozone therapy for 30 minutes, without pulsed electromagnetic field therapy) versus half of the cells places on room air temperature (control) in the same room where the HOCATT machine was located. Real time-polymerase chain reaction was performed on both groups of cells and the results showed that cells placed in the HOCATT showed a significant 5 times increase in CYP19A1 mRNA levels and 50% significant decrease in SCC mRNA levels (*P < 0.05 for both). Data are presented as mean ± SEM, and were analyzed by paired t-test. The detailed statistical analyses are shown in Additional Table 1.3B-HSD: 3 beta-hydroxysteroid dehydrogenase; SCC: side-chain cleavage enzyme, also called CYP11A1; StAR: steroidogenic acute regulatory protein.