Targeted Contrast Enhanced Ultrasound For In Vivo Quantitative Evaluation On Angiogenesis In Transplanted Ovarian Of Rats

Objective:To evaluate the value of targeted contrast enhanced ultrasound(CEUS)using anti-Mlleriancanal hormone(AMH)targeted nanobubbles(AMH-NB)for in vivo quantitative evaluation on ovarian neovascular density after ovarian antu-transplantation in SD rats.Methods:Use"thin film hydration method"and"avidin-biotin method"to prepare AMH-NB and test its basic physical properties.Transplant autologous ovaries into the right inguinal area to establish a rat autologous ovarian transplantation model.The targeted(AMH-NB),non-targeted(N-NB)contrast agents and Sono Vue were administered on the 7th days after transplantation to obtain Peak Intensity(PI)and Time To Peak(TTP).The microvascular density(MVD)was measured using immunohistochemistry,and the correlation of PI,TTP and MVD were analyzed.Analyze whether AMH-NB enters the interstitial space by immunofluorescence.Results:The particle size of AMH-NB uniformly distributed,ranged(622.67±33.65)nm,and the concentration of AMH-NB was(2.90±0.26)×10~8/m L.PI of ovarian angiography value with AMH-NB was(7.93±0.65)d B,and TTP was(42.53±1.74)s,with N-NB was(6.14±0.44)d B,and(54.35±1.73)s,with Sonovue was(4.15±0.83)d B,and(28.71±1.18)s,respectively(all P<0.05).Immunohistochemical analysis showed that the microvascular density was(61.20±6.84)/HP,histological analysis indicated that AMH-NB were able to penetrate blood vessels to the interstitial space and conbine with AMH.PI,TTP of AMH-NB were highly both correlated with MVD(r=0.84,r=-0.84,P<0.05).Sonovue group(r=0.67,r-=-0.71,P<0.05),N-NB group(r=0.65,r-=-0.68,P<0.05)PI and PPT were found in MVD respectively.Conclusion:AMH-NB can be used to qualitatively and quantitatively evaluate the angiogenesis in transplanted ovarian of rats in vivo with CEUS.