Endometrial microbiome: sampling, assessment, and possible impact on embryo implantation

Abstract

There is growing interest on the potential clinical relevance of the endometrial microbiome. However, insufficient attention has been given to the methodology of sampling. To minimize contamination, we advocate the use of the double-lumen catheters commonly employed for the embryo transfer. Endometrial fluid samples obtained from 53 women scheduled for IVF were studied for microbiome characterization. Control samples from the vagina of these same women were concomitantly obtained. Samples were analysed by V3-V4-V6 regions of 16S rRNA gene sequencing with Next Generation Sequencing technique. Endometrial Lactobacillus-dominant cases were uncommon compared to previous evidence, being observed in only 4 (8%) women. Taxonomy markedly differed between the endometrial and vaginal microbiomes composition. The most common bacterial genera coincided in only 4 (8%) women. The comparison between women who did and did not subsequently become pregnant failed to identify any microorganism associated with the success of the procedure. However, the endometrial biodiversity resulted higher among pregnant women. Shannon's Equitability index in pregnant and non pregnant women was 0.76 [0.57-0.87] and 0.55 [0.51-0.64], respectively (p = 0.002). In conclusion, the use of embryo transfer catheters for testing the endometrial microbiome is promising. The scant concordance with vaginal samples supports the validity of this approach. Moreover, our study highlighted a possible beneficial role of a higher biodiversity on endometrial receptivity.

Figure 1

The double-lumen catheter used for endometrial sample collection. This type of catheter is commonly used for embryo transfer. The outer hollow catheter (represented in grey) is placed with its distal part just above the internal os of the cervix. Thereafter, the inner catheter (with a smaller diameter, represented in white) can easily pass through the first one, avoiding contacts with the vaginal and cervical mucosae. Once reached the endometrial cavity, endometrial collection of the endometrial fluid can be obtained by gentle traction on the syringe. The box of upper left corner of the figure shows in more details the mechanism of the double-lumen system.

Figure 2

Shannon and Shannon’s equitability indexes in women who did and did not become pregnant. A statistically significant difference emerged for both indexes in the endometrial microbiomes (p = 0.036 and p = 0.002, respectively). Conversely, no significant differences could be observed for the vaginal microbiomes.