Human platelet-rich plasma improves endometrial regeneration and pregnancy outcomes in a murine model of asherman syndrome

Abstract

AS is characterized by intrauterine adhesion or fibrosis, which are usually sequelae from damage to the endometrium, and is often associated with infertility. There are few therapeutic options that are not practically effective. The efficacy of autologous PRP including several cytokines and growth factors was recently suggested for patients with impaired endometrium. We investigated whether PRP administration could restore endometrial function and increase pregnancy outcomes in a murine model of AS. Experimental study Under IRB approval, humans PRP were donated from 6 patients. We have established a murine model of AS by inducing damage to the uterine horns. The 35 immunodeficient mice were randomly assigned into sham, AS group. 0.02ml of PRP was administrated into the unilateral horn or bilateral horns of AS group by intrauterine injection at day 7 after injury. The cellular and molecular fibrosis was analyzed by histologic and immunofluorescence staining and comparative expression of fibrosis-related factors at day 7 after PRP treatment. Female mice were individually bred to males with proven fertility to evaluate embryos on day 12 and the live birth rate. The histologic evidence of regeneration in damaged horn after PRP treatment was confirmed by hematoxylin and eosin, and Mansson’s trichrome staining. The expression of COL1A1 was lower in the PRP-treated AS horn in comparison with the untreated AS horn on immunofluorescence staining. The result RT-PCR and real-time RT-PCR of fibrosis-related factors (Tgfβ1, Timp1, and Col1a1) revealed that intrauterine infusion of PRP was effective to recuperate traumatized horn (5.9 vs. 2.6; 3.1 vs. 1.0; 4.4 vs. 1.0, p<0.01). The number of implantation site of AS horn was improved by PRP treatment (2.1 vs. 4.6, p<0.01). The live birth rate in the untreated AS group and in the PRP treated AS group was 0% and 83.3%, respectively (p<0.01). On average, litter size were 6.3 in the PRP-treated AS group and 11 in the sham group (p<0.01). The weight of the pups born to PRP-treated mice was similar to that of sham mice on day 7 to 28 after birth. This is the first study so far to show that human PRP is able to regenerate impaired endometrium and to enhance pregnancy outcomes in a murine model of AS. In previous studies, endometrial development was assessed by ultrasound to confirm the effect of PRP treatment in humans. These data provide evidence for histological alteration and changed expression of fibrosis-related factors. We believe that our studies support the potential value of using PRP for endometrial regeneration in clinical settings with compromised endometrial growth.