Autologous platelet-rich plasma dual-network hydrogel promotes human endometrial regeneration

Intrauterine perfusion of platelet-rich plasma (PRP) is often used for the treatment of intrauterine adhesions (IUA). However, the further application of PRP is limited by its difficulty in attaching to the endometrial surface and its inability to release growth factors persistently. Therefore, in this study, a dual-network (DN) hydrogel was prepared by using hyaluronate gel as the matrix and cross-linked PRP, which was applied to the clinical treatment of IUA. The DN hydrogel showed non-swelling in PBS, and 28 % of DN hydrogel remained after 96 h in vivo, in which PDGF-BB and VEGF showed sustained linear releasement. Also, DN hydrogel could promote the proliferation of endometrial epithelial cells in human endometrial epithelial organoids, suggesting that PRP releasing PDGF-BB and VEGF in DN hydrogel played a role. After treatment of DN hydrogel in patients with moderate to severe IUA, the menstrual volume and subendometrial blood flow were significantly increased, the IUA score was significantly decreased, the number of endometrial epithelial glands was increased, and the expression levels of ERα, MKI67 and CD31 were increased after treatment. Further single-cell sequencing analysis of the endometrium before and after DN hydrogel treatment revealed that WNT signaling pathway was significantly enriched after treatment. Immunohistochemical results confirmed that the increased expression of SOX9 and LGR5 in human endometrial glands after treatment, and also DN hyrdrogel promoted SOX9 and LGR5 expression in organoids. In conclusion, this DN hydrogel can improve the therapeutic effect of PRP and promote endometrial repair and regeneration in patients. STATEMENT OF SIGNIFICANCE: Dual-network hydrogels combined with Platelet-rich plasma (PRP) were successfully applied to human intrauterine adhesions and achieved the desired therapeutic effect. The PRP dual-network hydrogel enhanced the mechanical properties of PRP and promoted endometrial repair and regeneration in clinical patients probably by activating the WNT signaling pathway. This study proposes a clinically practical therapeutic strategy and provides a foundation for the utilization of PRP in the field of endometrial regeneration.