Anti-inflammatory drugs disrupt granulosa cell homeostasis in vitro and impair bovine oocyte maturation and embryo development

Granulosa cells (GCs) play a crucial role in maintaining follicular homeostasis and supporting the acquisition of oocyte developmental competence. Disruption of GC function may therefore impair oocyte maturation and subsequent embryo development. Although anti-inflammatory drugs are widely used in cattle, their potential direct effects on follicular function and reproductive competence remain poorly understood. The aim of this study was to evaluate whether exposure of bovine GCs to non-steroidal anti-inflammatory drugs (NSAIDs) and a steroidal anti-inflammatory drug (SAID) affects GC viability and morphology, and whether these alterations influence oocyte maturation and embryo developmental competence in vitro. Bovine GCs were exposed to Meloxicam, Flunixin, and Ketoprofen (NSAIDs) or Dexamethasone (SAID) at different concentrations and exposure times. Cell viability and morphology following treatment were evaluated by acridine orange and MTT test. Functional consequences were investigated using a GC-oocyte transwell co-culture system to assess oocyte maturation through first polar body extrusion and subsequent embryo development in vitro . Dexamethasone induced the most pronounced alterations in GC morphology and monolayer integrity. All drug treatments significantly reduced (P < 0.05) the percentages of matured oocytes and blastocyst formation on day 7 compared with controls. On day 11 of embryo culture, Flunixin (5 μM, 8 h) and Ketoprofen (10 μM, 48 h) exerted the most severe effects, as no hatched blastocysts were observed. GC alterations were associated with impaired oocyte developmental competence and reduced embryo developmental potential. These findings suggest that exposure to anti-inflammatory drugs disrupts GC function and impairs oocyte developmental competence and embryo development in vitro .