Development and characterization of a chick embryo chorioallantoic membrane (CAM) based platform for evaluation of
Lei Chen1, Shuncong Wang1, Yuanbo Feng1
1KU Leuven, Biomedical Group, Campus Gasthuisberg, Leuven 3000, Belgium.
Abstract:
Among various anti-cancer therapies, tumor vascular disrupting agents (VDAs) play a crucial role, for which their off-targeting effects on normal vessels need also to be investigated. The purpose of this study was to set up an in-ovo platform that combines a laser speckle contrast imaging (LSCI) modality with chick embryo chorioallantoic membrane (CAM) to real-time monitor vascular diameters and perfusion without and with intravascular injection. Two eggshell windows for both observation or measurement and injection were opened. Dynamic blood perfusion images and corresponding statistic graphs were acquired by using a LSCI unit on CAMs from embryo date (ED) 9 to ED15. A dedicated fine needle catheter was made for slow intravascular administration over 30 min with simultaneous LSCI acquisition. To verify the connectivity between CAM vessels and the embryonic circulations in the egg, contrast-enhanced 3D micro computed tomography (μCT), 2D angiography and histology were executed. This platform was successfully established to acquire, quantify and demonstrate vascular and hemodynamic information from the CAM. Chick embryos even with air cell opened remained alive from ED9 to ED15. Through collecting LSCI derived CAM vascular diameter and perfusion parameters, ED12 was determined as the best time window for vasoactive drug studies. A reverse correlation between CAM vessel diameter and blood perfusion rate was found (p < 0.002). Intravascular infusion and simultaneous LSCI acquisition for 30 min in ovo proved feasible. Contrast-enhanced angiography and histomorphology could characterize the connectivity between CAM vasculature and embryonic circulation. This LSCI-CAM platform was proved effective for investigating the in-ovo hemodynamics, which paves the road for further preclinical research on vasoactive medications including VDAs.
Insights
Researchers developed an in-ovo platform using laser speckle contrast imaging (LSCI) on chick embryo membranes to study blood flow and vessel changes. This method is effective for preclinical research on anti-cancer drugs like vascular disrupting agents (VDAs).
Area of Science:
- Cardiovascular Research
- Developmental Biology
- Pharmacology
Background:
- Tumor vascular disrupting agents (VDAs) are vital anti-cancer therapies, necessitating investigation into their effects on normal vasculature.
- Assessing off-target effects of VDAs on healthy blood vessels requires robust preclinical models.
Purpose of the Study:
- To establish an in-ovo platform integrating laser speckle contrast imaging (LSCI) with the chick embryo chorioallantoic membrane (CAM) model.
- To enable real-time monitoring of vascular diameters and blood perfusion in CAM vasculature.
- To facilitate the study of vasoactive drug effects, including VDAs, in a live embryonic system.
Main Methods:
- Developed an in-ovo platform using LSCI on chick embryo CAM from embryonic day (ED) 9 to ED15.
- Utilized eggshell windows for observation, measurement, and intravascular injection.
- Administered agents via a fine needle catheter over 30 minutes with simultaneous LSCI acquisition.
- Verified CAM vasculature and embryonic circulation connectivity using contrast-enhanced 3D micro-CT, 2D angiography, and histology.
Main Results:
- Successfully established a platform for acquiring and quantifying vascular and hemodynamic data from CAM.
- Demonstrated chick embryo survival from ED9 to ED15 with the experimental setup.
- Identified ED12 as the optimal window for vasoactive drug studies based on LSCI data.
- Observed a significant inverse correlation between CAM vessel diameter and blood perfusion rate (p < 0.002).
- Confirmed the feasibility of in-ovo intravascular infusion with simultaneous LSCI.
Conclusions:
- The LSCI-CAM platform is effective for investigating in-ovo hemodynamics and vascular responses.
- This model provides a valuable tool for preclinical research on vasoactive medications, including VDAs.
- The study highlights the potential for real-time hemodynamic assessment in embryonic models.
