Xin-Min Tang

Postsurgical intraperitoneal exposure to glove powders modulates inflammatory and immune‐related cytokine production

The objective of the present study was to determine whether intraperitoneal exposure to glove powders modulates the inflammatory and immune responses by altering the influx of inflammatory and immune cells and peritoneal fluid cytokines and thus the outcome of surgically induced peritoneal wound healing. Peritoneal wall injuries were made by scraping the tissue until bleeding occurred in 360 mice. One of the following fluids was then introduced into the peritoneal cavity: phosphate‐buffered saline solution, phosphate‐buffered saline solution containing glove powders (Biosorb and Keoflo, 100 µg/ml), Hydrocote (Hydrogel film, Biogel 100 µg/ml), latex proteins (1 mg/ml), or lipopolysaccharides (12.5 µg/ml). At intervals of 1 to 28 days after injury, 10 mice per treatment per day and 10 uninjured mice were killed, peritoneal fluids were collected to determine the cytokine levels, the rate of fibrous adhesions formed at the site of injuries was graded, and peritoneal walls with attached fibrous adhesions were removed to determine the degree of inflammatory and immune cell infiltration into the wound. The results indicated that, with the exception of interferon‐γ, the peritoneal fluid levels of transforming growth factor‐β1, tumor necrosis factor‐α, interleukin‐1β, and granulocyte‐macrophage‐colony stimulating factor in the phosphate‐buffered saline solution‐treated injured group significantly increased, reaching maximum between days 4 and 7 (p < 0.05) compared with the uninjured group and returned to uninjured values by day 14 after injury. The level of transforming growth factor‐β1 was higher in glove powders and Hydrocote‐treated groups than in latex, liposaccharides, or phosphate‐buffered saline solution‐treated groups until day 14 after surgery (p < 0.05). The levels of tumor necrosis factor‐α and interleukin‐1β increased in all treatment groups during the first week after injury compared with uninjured controls, with the exception of Hydrocote. The number of T helper/inducers (CD4), total leukocytes (CD11a), B lymphocytes (CD45R), granulocytes (Gr‐1), and mononuclear phagocytes (Mac‐3) in the wound increased during the first week after peritoneal wounding with no significant difference between treated and untreated groups. The rate of adhesion formation was not significantly altered in treated compared with untreated groups. These data suggest that a mechanism which mediates glove powder‐induced peritoneal inflammatory and immune reactions in the postsurgical setting involves augmentation of cytokine production without influencing the influx of inflammatory and immune cells or adhesion formation.

Modulation of mouse sperm-egg interaction, early embryonic development and trophoblastic outgrowth by activated and unactivated macrophages.

Exposure of mouse spermatozoa and oocytes duringin vitro fertilization (IVF) to lipopolysaccharide (LPS) and phorbol myristate acetate (PMA) activated macrophages (U937 cell line), but not unactivated macrophages cultureconditioned medium or control medium (RMPI+DMEM with 0.5% FBS) resulted in inhibition of IVF (87.2%), first cleavage (90.8%) and total blastocyst formation 97.5%). The direct coculture of the activated macrophages with 2-cell stage embryos resulted in arrested development (91.2%), an effect that was significantly diminished in the presence of monolayer of human endometrial stromal cells in the coculture (58.3%). In contrast, the majority of 2-cell embryos developed to blastocysts when exposed to unactivated macrophages, or macrophage-stromal cell cocultures (94.1%). The majority of 2-cell embryos cultured in control medium (DMEM/Ham’s F12 with 2% FBS) developed to morulae (96.2%), then underwent growth arrest and degeneration. Furthermore, culturing blastocyst stage embryos in the above groups resulted in a significant enhancement of trophoblast outgrowth, particularly in coculture with activated macrophages as compared to any other group (P<0.005). There was a significant increase in the levels of TGF-β, GM-CSF, IL-1α, IL-1β, TNF-α, PGE2, TXB2 and LTB4 released into the culture conditioned medium of activated macrophages compared to unactivated macrophages (P<0.001). These results suggest that the secretory products of activated macrophages, among them those determined in this study, in a stage-specific manner can directly effect sperm-egg interaction, early embryonic development and trophoblastic outgrowth. This data provides further support for the hypothesis that in endometriosis-associated infertility, continuous exposure of spermatozoa, oocytes and early embryos to activated macrophage-derived factors may play a vital role in their survival during transportation and fertilization as well as development during early embryonic stage.

Surgical glove powders differentially modulate macrophage and lymphocyte-derived cytokines and eicosanoids production in vitro.

The objective of the present study was to determine the effect of surgical glove powders Biosorb, Keoflo, and CaCO3 and Hydrocote (a powder‐free film; Biogel) on cytokine and eicosanoid production by lipopolysaccharide/phorbol 12‐myristate 13‐acetate activated and unactivated HL60, U937, and RPMI 1788 cells, human monocyte/macrophage, and B lymphocyte cell lines. The unactivated cell culture—conditioned media contained a low level of interleukin‐1α and ‐1β, granulocyte macrophage colony stimulating factor, and tumor necrosis factor‐α, which significantly increased after activation (p < 0.05). Exposure of unactivated cells to glove powders or Hydrocote (100 µg/ml) had little effect. However, these compounds appeared to have multiple inhibitory and stimulatory action on the production of these cytokines and eicosanoids in lipopolysaccharide/phorbol 12‐myristate 13‐acetate activated cells. For instance, granulocyte macrophage colony stimulating factor production was inhibited only in U937 cells by Keoflo and CaCO3, whereas, tumor necrosis factor‐α production was stimulated by Biosorb and Keoflo in HL‐60, and CaCO3 was found to be predominantly inhibitory on tumor necrosis factor‐α production by these cells (p < 0.05). Total transforming growth factor‐β1 production was stimulated by Biosorb and Hydrocote in U937 and HL‐60 cells, respectively, but inhibited by Keoflo in U937 cells. However, Biosorb and Keoflo inhibited transforming growth factor‐β1 production in both HL‐60 and RPMI 1788 cells, without any effect on active transforming growth factor‐β1. With regard to eicosanoids, Biosorb and Keoflo stimulated prostaglandin E2 production by RPMI 1788 cells, whereas it was inhibited by all glove powders in HL‐60 cells. Thromboxane B2 production was stimulated by Keoflo and inhibited by CaCO3 and Hydrocote in U937 cells. Finally, Leukotriene B4 synthesis was found to become stimulated by Keoflo, CaCO3, and Hydrocote in both HL60 and RPMI 1788 cells (p < 0.05). These data indicate that exposure of activated, but not unactivated, macrophages and lymphocyte to surgical glove powders and Hydrocote differentially effects the release of cytokines and eicosanoids by these cells. Considering that cytokines and eicosanoids play an important role in mediating the inflammatory and immune responses of wound healing, complications arising from glove powder exposure in vivo may involve mechanisms which alter the type and level of cytokine and eicosanoid production.