| Popis: |
Despite the presence of an intense research in the field of the equine reproduction over recent decades, there has been no significant increase in the population's fertility. To date, few therapeutic protocols are available for the treatment of equine reproductive disorders because of the peculiarities of species but also for the ethical, clinical and economical reasons adressed to the in vivo tests. An easily obtainable sample for performing reliable in vitro tests is the endometrial biopsy; however, the lack of studies regarding the maintenance of the vitality for this sample is still an obstacle. For this purpose, the first aim of the present work has been the creation of a protocol for the culture of equine endometrial biopsies; moreover, we assessed the survival of two different kind of biopsy (incisional and excisional) by the estimation of mitochondrial activity, DNA quantity and the histology of the samples at 3 and 7 days of in vitro culture at 37°C and 5% CO2. Results show that incisional biopsies maintain an adequate level of vitality up to 3 days, representing a valid sample to be used for in vitro testing. In addition, the method allows the evaluation of innovative therapies such as mesenchymal stem cells (MSCs). MSCs have been shown to support tissue regeneration and to modulate inflammatory processes. For these reasons, MSCs could be used in equine endometritis and/or endometrosis, inflammatory-based pathologies that represent one of the major reproductive diseases for horse because of reducing the fertility of the animal and increasing costs for breeders. However, further insights are needed to improve the knowledge about their mechanism of action and their practical use. On the light of that, the second aim of this project has been the evaluation of the expression of genes related to the production of inflammatory cytokines after co-culture for 3 days of incisional endometrial biopsies with two different doses of adipose derived stem cells (ADSCs) (1x10^5 and 3 x 10^5 cells). The preliminary results allow us to hypothesize that the presence of ADSCs modifies the expression of genes involved in the production of cytokines with an immunomodulatory effect and possible activation of anti-bacterial and anti-fibrotic mechanisms that could be the starting point for the restoration of damaged endometrium. In addition, for some cytokines this immunomodulatory effect seems to be dose-dependent and deserves further studies. |
