title: Drug delivery to the brain by focused ultrasound induced blood-brain barrier disruption: quantitative evaluation of enhanced permeability of cerebral vasculature using two-photon microscopy.
abstract: Reversible and localized blood-brain barrier disruption (BBBD) using focused ultrasound (FUS) in combination with intravascularly administered microbubbles (MBs) has been established as a non-invasive method for drug delivery to the brain. Using two-photon fluorescence microscopy (2 PFM), we imaged the cerebral vasculature during BBBD and observed the extravasation of fluorescent dye in real-time in vivo. We measured the enhanced permeability upon BBBD for both 10 kDa and 70 kDa dextran conjugated Texas Red (TR) at the acoustic pressure range of 0.2-0.8 MPa and found that permeability constants of TR10 kDa and TR70 kDa vary from 0.0006 to 0.0359 min(-1) and from 0.0003 to 0.0231 min(-1), respectively. For both substances, a linear regression was applied on the permeability constant against the acoustic pressure and the slope from best-fit was found to be 0.039 ± 0.005 min(-1)/MPa and 0.018 ± 0.005 min(-1)/MPa, respectively. In addition, the pressure threshold for successfully induced BBBD was confirmed to be 0.4-0.6MPa. Finally, we identified two types of leakage kinetics (fast and slow) that exhibit distinct permeability constants and temporal disruption onsets, as well as demonstrated their correlations with the applied acoustic pressure and vessel diameter. Direct assessment of vascular permeability and insights on its dependency on acoustic pressure, vessel size and leakage kinetics are important for treatment strategies of BBBD-based drug delivery.
authors.affiliation: Physical Sciences, Sunnybrook Research Institute, Toronto, ON, Canada; Medical Biophysics, University of Toronto, Toronto, ON, Canada. Electronic address: tnhan@sri.utoronto.ca.
authors.forename: Tam
authors.initials: T
authors.lastname: Nhan
authors.affiliation: Physical Sciences, Sunnybrook Research Institute, Toronto, ON, Canada.
authors.forename: Alison
authors.initials: A
authors.lastname: Burgess
authors.affiliation: Physical Sciences, Sunnybrook Research Institute, Toronto, ON, Canada.
authors.forename: Eunice E
authors.initials: EE
authors.lastname: Cho
authors.affiliation: Physical Sciences, Sunnybrook Research Institute, Toronto, ON, Canada; Medical Biophysics, University of Toronto, Toronto, ON, Canada.
authors.forename: Bojana
authors.initials: B
authors.lastname: Stefanovic
authors.affiliation: Medical Biophysics, University of Toronto, Toronto, ON, Canada; Ontario Cancer Institute, Princess Margaret Hospital, Toronto, ON, Canada.
authors.forename: Lothar
authors.initials: L
authors.lastname: Lilge
authors.affiliation: Physical Sciences, Sunnybrook Research Institute, Toronto, ON, Canada; Medical Biophysics, University of Toronto, Toronto, ON, Canada.
authors.forename: Kullervo
authors.initials: K
authors.lastname: Hynynen
mesh_terms: D000818:Animals; D001812:Blood-Brain Barrier / Q000378:metabolism*; D001921:Brain / Q000098:blood supply*; D002199:Capillary Permeability; D003911:Dextrans / Q000008:administration & dosage*; D016503:Drug Delivery Systems / Q000295:instrumentation*; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000008:administration & dosage*; D008297:Male; D036641:Microscopy, Fluorescence, Multiphoton; D051381:Rats; D017208:Rats, Wistar; D014465:Ultrasonics / Q000295:instrumentation*; D014966:Xanthenes / Q000008:administration & dosage*
chemical_list: D003911:Dextrans; D005456:Fluorescent Dyes; D014966:Xanthenes; C034657:Texas red
keywords: Blood–brain barrier; Drug delivery; Focused ultrasound; Permeability; Two-photon fluorescence microscopy