Abstract
Title: A Novel Tissue Explant Culture Model For Testing of Candidate Treatments for Inflammation of the Uterus in Pigs.
Keywords: Endometritis; Inflammation; Treatments;
Authors: Joanna Giles, Robert Walton, Ruth Wonfor* and Debbie Nash*
Background: Endometritis is an inflammatory condition of the uterus leading to complications with the reproductive organs, miscarriage and infertility. Persistent endometritis in livestock animals is an ongoing problem for producers, costing the UK an estimated £110 million in treatments annually.
The first-line treatment of endometritis in pigs, cows and horses are antibiotics, which do not always resolve inflammation and heavily contribute to antibiotic resistance. As such, there is an urgent need for the development of alternative treatments for endometritis. Here we propose, optimise, and validate the first porcine tissue explant model for uterine inflammation and test naturally derived compounds for their effectiveness as anti-inflammatory therapies in porcine endometrial tissue.
Methods: Uterine endometrial explants were harvested from uteri of sows sent to slaughter. Tissue biopsies were collected at follicular (n=4) and luteal phases (n=6) of the oestrous cycle. Uteri were opened using a sterile scalpel and the endometrium layer biopsied and dissected from the myometrium. Biopsies were washed and each placed into wells of a 6-well culture plate. These tissue ‘explants’ were maintained in 3mL of supplemented DMEM-F12 at 37℃ and 5% CO₂ for 18 hr. Explants were then cultured for 24 hours with oxytocin (to stimulate PGF2α production) or LPS to mimic infection and induce an inflammatory response. The explants were weighed, and tissues retained for RT-PCR analysis. Supernatants were harvested and concentrations of markers were measured by ELISA and Luminex.
Results: The data generated thus far show a dose dependent response of IL-6 secretion to LPS stimulation, which demonstrates the validity of our model. We show that inflammation can be stimulated in an in vitro setting to mimic porcine endometritis and present data to show the change in interleukin secretion in response to treatment with naturally derived compounds.
Conclusions: The data we present show that inflammation can be stimulated in an in vitro setting to mimic porcine endometritis The explant model is a useful tool for further testing of novel compounds and their potential anti-inflammatory effects on the porcine endometrium.
Keywords: Endometritis; Inflammation; Treatments;
Authors: Joanna Giles, Robert Walton, Ruth Wonfor* and Debbie Nash*
Background: Endometritis is an inflammatory condition of the uterus leading to complications with the reproductive organs, miscarriage and infertility. Persistent endometritis in livestock animals is an ongoing problem for producers, costing the UK an estimated £110 million in treatments annually.
The first-line treatment of endometritis in pigs, cows and horses are antibiotics, which do not always resolve inflammation and heavily contribute to antibiotic resistance. As such, there is an urgent need for the development of alternative treatments for endometritis. Here we propose, optimise, and validate the first porcine tissue explant model for uterine inflammation and test naturally derived compounds for their effectiveness as anti-inflammatory therapies in porcine endometrial tissue.
Methods: Uterine endometrial explants were harvested from uteri of sows sent to slaughter. Tissue biopsies were collected at follicular (n=4) and luteal phases (n=6) of the oestrous cycle. Uteri were opened using a sterile scalpel and the endometrium layer biopsied and dissected from the myometrium. Biopsies were washed and each placed into wells of a 6-well culture plate. These tissue ‘explants’ were maintained in 3mL of supplemented DMEM-F12 at 37℃ and 5% CO₂ for 18 hr. Explants were then cultured for 24 hours with oxytocin (to stimulate PGF2α production) or LPS to mimic infection and induce an inflammatory response. The explants were weighed, and tissues retained for RT-PCR analysis. Supernatants were harvested and concentrations of markers were measured by ELISA and Luminex.
Results: The data generated thus far show a dose dependent response of IL-6 secretion to LPS stimulation, which demonstrates the validity of our model. We show that inflammation can be stimulated in an in vitro setting to mimic porcine endometritis and present data to show the change in interleukin secretion in response to treatment with naturally derived compounds.
Conclusions: The data we present show that inflammation can be stimulated in an in vitro setting to mimic porcine endometritis The explant model is a useful tool for further testing of novel compounds and their potential anti-inflammatory effects on the porcine endometrium.
| Original language | English |
|---|---|
| Pages | 224.09 |
| Publication status | Published - 01 May 2023 |