Regenerative Potential of EO and HEO Complexes on Endometrial Injury and Vascular Formation in a Murine Model

Evaluation of EO and HEO Complex Efficacy in Endometrial Regeneration and Vascular Formation Using a Murine Model

This study aimed to investigate the therapeutic potential of EO and HEO complexes in endometrial regeneration and vascular formation. We employed a two-pronged approach using a murine model:

1. Endometrial Injury Model:

• Animals: Female SCID mice aged 6-8 weeks were utilized.* Endometrial Injury Induction: Following anesthesia, a laparotomy was performed to expose the left uterus. Ligatures were placed at the oviductal and cervical ends of the uterine horns. Subsequently, 95% ethanol was injected into the uterine cavity via the oviductal end, followed by a 15-second pause and PBS flushing.* Treatment Groups: * EO complex group: Mice received 75 μl of EO complex cultured in eCHIP with estradiol for 2 days, injected into the injured uterine cavity. * HEO complex group: Mice received 75 μl of HEO complex cultured in eCHIP with estradiol for 2 days, injected into the injured uterine cavity. * Matrigel group: Mice received an equal volume of Matrigel. * Natural repair group: Mice received an equal volume of PBS. * Post-Operative Procedures: After 3 minutes, ligatures were removed, and the uterus was repositioned and sutured.* Endpoint Analysis: * Mice were euthanized at 3 and 7 days post-surgery for uterine collection and subsequent pathological and immunohistochemical staining. * Fertility assessment was conducted 14 days post-surgery by housing the mice with reproductively active males.

2. Vascular Formation Assessment:

• Transplantation: 30 μl of either EO or HEO complex was transplanted subcutaneously on the dorsum of female SCID mice.* Angiogenesis Visualization: After 14 days, fluorescent substances were injected intravenously. After a 30-minute interval, the transplants were excised and examined under a stereomicroscope for confocal microscopy fluorescence imaging to assess vascular formation.

Significance: This study provides valuable insights into the potential of EO and HEO complexes in promoting endometrial repair and neovascularization, which could have significant implications for treating female infertility and other reproductive disorders.