Blood-borne brain metastases are associated with poor prognosis, but
little is known about the interplay between cerebral blood flow, surgical stress
responses, and the metastatic process. The intra-carotid inoculation approach,
traditionally used in animal studies, involves permanent occlusion of the common
carotid artery (CCA). Herein we introduce a novel intra-carotid inoculation
approach that avoids CCA ligation, namely – assisted external carotid
artery inoculation (aECAi) – and compared it to the traditional approach
in C57/BL6 mice, assessing cerebral blood flow; particle distribution;
blood-brain barrier (BBB) integrity; stress, inflammatory and immune responses;
and tumor brain retention and growth. Doppler flowmetry and two-photon imaging
confirmed that only in the traditional approach regional and capillary cerebral
blood flux were significantly reduced. Corticosterone and plasma IL-6 levels
were higher in the traditional approach, splenic numbers of NK, CD3+,
granulocytes, and dendritic cells were lower, and many of these indices were
more profoundly affected by surgical stress in the traditional approach. BBB
integrity was unaffected. Administration of spherical beads indicated that CCA
ligation significantly limited brain distribution of injected particles, and
inoculation of D122-LLC syngeneic tumor cells resulted in 10-fold lower brain
tumor-cell retention in the traditional approach. Last, while most of the
injected tumor cells were arrested in extra-cranial head areas, our method
improved targeting of brain-tissue by 7-fold. This head versus brain
distribution difference, commonly overlooked, cannot be detected using
in vivo bioluminescent imaging. Overall, it is crucial to
maintain unperturbed cerebral blood flow while studying brain metastasis and
interactions with stress and inflammatory responses.