Stretch-induced intussuceptive and sprouting angiogenesis in the chick chorioallantoic membrane

• A defined stretch field was applied to the chick chorioallantoic membrane.
• Stretch-induced angiogenesis demonstrated a marked increase in vessel density.
• Conducting vessels demonstrated axial growth and diminished bifurcation angles.
• The capillary network demonstrated dramatic intussusceptive and sprouting angiogenesis.

Vascular systems grow and remodel in response to not only metabolic needs, but also mechanical influences as well. Here, we investigated the influence of tissue-level mechanical forces on the patterning and structure of the chick chorioallantoic membrane (CAM) microcirculation. A dipole stretch field was applied to the CAM using custom computer-controlled servomotors. The topography of the stretch field was mapped using finite element models. After 3 days of stretch, Sholl analysis of the CAM demonstrated a 7-fold increase in conducting vessel intersections within the stretch field (p < 0.01). The morphometric analysis of intravital microscopy and scanning electron microscopy (SEM) images demonstrated that the increase vessel density was a result of an increase in interbranch distance (p < 0.01) and a decrease in bifurcation angles (p < 0.01); there was no significant increase in conducting vessel number (p > 0.05). In contrast, corrosion casting and SEM of the stretch field capillary meshwork demonstrated intense sprouting and intussusceptive angiogenesis. Both planar surface area (p < 0.05) and pillar density (p < 0.01) were significantly increased relative to control regions of the CAM. We conclude that a uniaxial stretch field stimulates the axial growth and realignment of conducting vessels as well as intussusceptive and sprouting angiogenesis within the gas exchange capillaries of the ex ovo CAM.