Purpose of the work. A post-mating-induced endometritis is a physiological self-limited and necessaryreaction to clear the uterus after insemination.1 Recently, platelet rich-plasma (PRP) wasshown effective in the treatment of chronic endometritis2 and in modulating inflammatory uterineresponse3 by production of growth factors, chemokines and cytochines. Aim of this study was toevaluate the effect of intrauterine autologous PRP infusion on the post- insemination inflammatoryendometrial response and on the pregnancy rate in sub-fertile mares inseminated withcooled-semen.Materials and methods. PRP treatment was performed in 11 sub-fertile mares [median age 11;range 7-19] unresponsive to conventional therapies. Artificial inseminations (AI) were performed 8or 24 hours after induction of ovulation with hCG (2000UI). A dose of 800-1200 x 106of cooled semendiluted (50 x 106 sperms/ml)with a milk based commercial extender was deposited in the uterinebody. At this moment, 300 ml of blood were collected from the jugular vein in blood bags containingCPDA-1. PRP was prepared with the method of double centrifugation2-4: blood was centrifugatedat 120 g x 20 min and the all the plasma was pipetted in new tubes to further centrifugationat 240x g for 10 min. Subsequently, the supernatant was discarded, and the remaining fractionwas used as PRP. Low volume flushes (LVF) with 120 ml of sterile saline were performed 6-8hours after AI and immediately after 20 ml of autologous PRP containing on average of 196000platelets/μL, were infused into the uterus. At 36h from the hCG administration, mares were checkedfor ovulation and a second LVF was performed. The turbidity of LVF samples were scored with a0-5 scale and then divided in two aliquots, one of which was immediately frozen at -20°C for furtherevaluation of inflammation markers and the other one was centrifuged at 240xg for 10 min. Thepellet was smeared and the number of neutrophils (PMN) and epithelial cells (EP) were counted in5 fields at 400x magnification.Outcomes. Data were checked for normality with Shapiro-Wilk test. EP and PMN cells and theratios PMN/EP pre and post PRP infusions were compared by Wilcoxon signed-rank test. The turbidityscores were compared by Mood median test. Differences were considered significant atp<0.05. Turbidity of the LVF fluid was significantly lower post PRP infusion (p=0,0002), scored4 (median; range 3-5) and 2 (range 1-3) respectively pre and post. PMN were 171.07 ±276.53(median ± dev st) and 12.57±35.02, EP were 6.67±4.2 and 79.71±115.34 and PMN/EP were21.29±29.75 and 0.39 ±1.03 respectively pre and post PRP infusion. All the differences werehighly significant (p=0.0006; p=0.0022; p=0.0002). Pregnancy rate at the first cycle post treatmentwas 63%.31823° CONGRESSO INTERNAZIONALE SIVEConclusion. Preliminary results show that PRP infusions post insemination could be effective inmodulating uterine inflammatory response to semen and improving pregnancy rate in subfertilemares.
Post-insemination intrauterine infusion of Platelet-Rich Plasma (PRP) in sub-fertile mares: preliminary results
C. Del Prete
;
2017-01-01
Abstract
Purpose of the work. A post-mating-induced endometritis is a physiological self-limited and necessaryreaction to clear the uterus after insemination.1 Recently, platelet rich-plasma (PRP) wasshown effective in the treatment of chronic endometritis2 and in modulating inflammatory uterineresponse3 by production of growth factors, chemokines and cytochines. Aim of this study was toevaluate the effect of intrauterine autologous PRP infusion on the post- insemination inflammatoryendometrial response and on the pregnancy rate in sub-fertile mares inseminated withcooled-semen.Materials and methods. PRP treatment was performed in 11 sub-fertile mares [median age 11;range 7-19] unresponsive to conventional therapies. Artificial inseminations (AI) were performed 8or 24 hours after induction of ovulation with hCG (2000UI). A dose of 800-1200 x 106of cooled semendiluted (50 x 106 sperms/ml)with a milk based commercial extender was deposited in the uterinebody. At this moment, 300 ml of blood were collected from the jugular vein in blood bags containingCPDA-1. PRP was prepared with the method of double centrifugation2-4: blood was centrifugatedat 120 g x 20 min and the all the plasma was pipetted in new tubes to further centrifugationat 240x g for 10 min. Subsequently, the supernatant was discarded, and the remaining fractionwas used as PRP. Low volume flushes (LVF) with 120 ml of sterile saline were performed 6-8hours after AI and immediately after 20 ml of autologous PRP containing on average of 196000platelets/μL, were infused into the uterus. At 36h from the hCG administration, mares were checkedfor ovulation and a second LVF was performed. The turbidity of LVF samples were scored with a0-5 scale and then divided in two aliquots, one of which was immediately frozen at -20°C for furtherevaluation of inflammation markers and the other one was centrifuged at 240xg for 10 min. Thepellet was smeared and the number of neutrophils (PMN) and epithelial cells (EP) were counted in5 fields at 400x magnification.Outcomes. Data were checked for normality with Shapiro-Wilk test. EP and PMN cells and theratios PMN/EP pre and post PRP infusions were compared by Wilcoxon signed-rank test. The turbidityscores were compared by Mood median test. Differences were considered significant atp<0.05. Turbidity of the LVF fluid was significantly lower post PRP infusion (p=0,0002), scored4 (median; range 3-5) and 2 (range 1-3) respectively pre and post. PMN were 171.07 ±276.53(median ± dev st) and 12.57±35.02, EP were 6.67±4.2 and 79.71±115.34 and PMN/EP were21.29±29.75 and 0.39 ±1.03 respectively pre and post PRP infusion. All the differences werehighly significant (p=0.0006; p=0.0022; p=0.0002). Pregnancy rate at the first cycle post treatmentwas 63%.31823° CONGRESSO INTERNAZIONALE SIVEConclusion. Preliminary results show that PRP infusions post insemination could be effective inmodulating uterine inflammatory response to semen and improving pregnancy rate in subfertilemares.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
