[Effect of contrast media used in hysterosalpingography (HSG) on phagocytosis of spermatozoa by peritoneal macrophages in female rats].

It is apparent from numerous communications that hysterosalpinogography (HSG), besides its diagnostic value, may also claim therapeutic activity. After such examinations, more frequent pregnancies were recorded in previously infertile women without resorting to supplementary treatment. Pregnancies were more numerous when applying oil-based contrast media than water-based contrast ones. Hypothesis has been put forward as to the possibility of modulating activity of these agents exerted upon the peritoneal and oviductal macrophages. Increased amount of these cells as well as excessive phagocytosis of spermatozoa was shown in case of endometriosis, one of the most frequent causes of infertility. It is supposed that excessive phagocytosis of spermatozoa by peritoneal macrophages (PM) may be the cause of infertility. Studies were performed to determine the effect of oil- and water-soluble contrast media, on sperm phagocytosis by PM in vitro. PM were obtained from Wistar female rats. Subsequently, the cells were centrifuged, washed and suspended in culture medium. Next 1 million PM were allowed to attach to the cover glass for one hour under standard conditions of incubation. Dilutions of 0.25%, 0.5%, 1.0% Lipiodol Ultra--Fluid (Byk Gulden Konstanz) and Uropolinum 60% (Polfa) were added to chambers with incubated PM for next one hour. The controls were cultured in the same conditions without contrast medium added. Sperm were isolated from the epididymal cauda of male rats and subsequently suspended in culture medium at a concentration 1 million/ml and added to equivalent volume of cultured macrophages for 1.5 hours. After the exposure time the cultures were washed and stained. The spermiophagic index (SPI) was determined. SPI = number of phagocytosed sperm cells/number of macrophages x 100. Statistical analysis was performed by means of Student t-test. Additional histochemical reactions were made and scanning as well as transmission electron microscopy studies were accomplished. The established results (Fig. 1-9) indicate that lipiodol inhibits phagocytosis of spermatozoa by PM stronger (1% lipodol SPI = 1.99 + 0.94 < 0.0001) than uropolinum (1% uropolinum SPI = 5.07 +/- 1.02 p < 0.0001). In control studies, without contrast medium added, SPI was equal to 14.66 +/- 3.12 (p < 0.00001). Marked inhibition involving phagocytosis of spermatozoa was detected already after the treatment with lipiodol in low concentration (0.25% lipiodol SPI = 3.73 +/- 0.89 p < 0.0001). It failed to be so distinct after treating macrophages with uropolinum in low concentration (0.25% uropolinum SPI = 8.34 +/- 1.50 p < 0.0001). Scanning electron microscopy studies have disclosed that in the cultures of macrophages with spermatozoa, the macrophages spread well over the glass bottom. The macrophages' cytoplasmic membrane was highly folded with numerous protrusions different in size and shape. They covered the digested parts of spermatozoon causing its fragmentation. In cultures of macrophages with spermatozoa and 1% uropolinum, the structure of macrophages was similar to that in the control group. Accumulations and conglomerations of a large number of macrophages with spermatozoa were additionally revealed. In cultures of macrophages with spermatozoa and 1% lipiodol, the macrophages were not spread, but rather spherical. Also the cytoplasmic membrane protrusions were small, short, fine and appeared in smaller number. Occasionally areas of smooth cytoplasmic membrane without any folds were seen. Such macrophages did not phagocytose sperm. Transmission electron microscopy studies of macrophages cultured with sperm have disclosed that the macrophages' cytoplasmic membrane was highly folded with numerous protrusions different in size and shape. The latter encircled and closed parts of sperm (head, midpiece or tail) inside inside endosomes, finally linking the latter with lysosomes. The ultrastructure of macrophages cultured with spermatozoa and 1% uropolinum was s