Temporal dynamics of neurovascular unit changes following blood-brain barrier opening in the putamen of non-human primates
Tiziano Balzano, José A. Pineda‐Pardo, Noelia Esteban-García, Miguel López‐Aguirre, Alejandro Reinares-Sebastián, Inés Trigo‐Damas, Masahiko Takada, José Á. Obeso, Javier Blesa
Abstract
Low-intensity focused ultrasound (LIFU) combined with intravenously circulating microbubbles has recently emerged as a novel approach for increasing delivery through the blood-brain barrier (BBB). This technique safely and transiently enables therapeutic agents to overcome the BBB, which typically poses a significant obstacle for treatment of brain disorders. However, the full impact of LIFU on the entire neurovascular unit (NVU), as well as the mechanisms and factors involved in restoring BBB integrity still require further elucidation. We conducted immunohistochemical analyses of the putamen in non-human primates to monitor changes over time [immediately post-treatment (3 h) and at 7- and 30-days post-BBB opening] in vascular, glial, and immune cells. Additionally, we examined the dynamic interactions among these elements and their role in the restorative process at the BBB level. A mild inflammatory response primarily involving microglia, astrocytes, and T- and B-lymphocytes was observed in the treated putamen acutely after BBB opening. These cells, recruited in response to the vascular changes, stimulate upregulation of PDGFRβ, a pericyte-specific marker, and VEGF-A, a pro-angiogenic factor. This was associated with vascular sprouting by 7 days post-BBB opening. Importantly, no notable long-term alterations were observed in the NVU 30 days post-BBB opening. These results offer further evidence regarding the efficacy and safety of LIFU in achieving BBB opening in the primate brain, indicating that nearly all changes in the NVU revert to baseline within 30 days post-treatment. This also suggests that angiogenesis may play an important role in restoring vascular integrity after BBB opening. • BBB permeability to Gd enhances up to 24 h post-LIFU, disappearing in 7 days. • LIFU-induced BBB opening triggers a mild, acute inflammatory response involving glia and lymphocytes. • PDGFRβ and VEGF-A upregulation leads to vascular sprouting within 7 days post-LIFU. • The neurovascular unit returns to baseline with no significant long-term changes by 30 days. • Angiogenesis plays a role in restoring vascular integrity after LIFU-induced BBB disruption.