Perivascular Delivery of Blebbistatin Reduces Neointimal Hyperplasia after Carotid Injury in the Mouse

Abstract

Proliferation and migration of smooth muscle cells (SMC) require myosin II activity; thus, we examined whether blebbistatin, a cell-permeable selective inhibitor of myosin II ATP activity, would impair neointimal hyperplasia after vascular injury. Delivery of blebbistatin via a perivascular polymer cuff reduced neointimal formation by 73% and luminal obstruction by 75% after carotid denudation injury in C57BL/6 mice. Blebbistatin treatment was also associated with a reduction in cell density within the neointima; total number of cells (76 Ϯ 7 to 27 Ϯ 3 cells/high-powered field) and actin-positive cells (64 Ϯ 4 to 24 Ϯ 2 cells/high-powered field) in the neointima were reduced in blebbistatin-treated mice compared with vehicle-treated mice. In a model of vascular injury with an intact endothelium, implantation of a blebbistatin-secreting cuff after carotid ligation in FVB/N mice was associated with a 61% decrease in neointimal area and a significant decrease in luminal obstruction (88 Ϯ 4% in vehicle-treated mice versus 36 Ϯ 4% in blebbistatin-treated mice; p Ͻ 0.0001). In cultured rat aortic SMC, blebbistatin disrupted cellular morphology and actin cytoskeleton structure, and these effects were rapid and completely reversible. Blebbistatin had a dose-dependent inhibitory effect on DNA replication and cell proliferative responses to platelet-derived growth factor-BB, angiotensin II, and ␣-thrombin, migratory responses to serum, and migratory responses after blunt injury. In summary, perivascular delivery of blebbistatin reduced neointimal hyperplasia after carotid injury in the mouse.

Actomyosin contractility provokes contact inhibition in E-cadherin-ligated keratinocytes

“…Blebbistatin treatment reduced cell proliferation in sparse cultures ( Fig. 1b ), which was consistent with the previous reports using different cell types 26 27 28 . By contrast, the ratio of EdU positive cells against total cells in confluent cultures was increased in response to the treatment with blebbistatin ( Fig.…”

Permeability mapping of gelatin methacryloyl hydrogels

“…Cell culture medium (Dulbecco’s Modified Eagle Medium, Sigma-Aldrich) was perfused within the straight channels of 500-μm diameter at a continuous rate of 20 μL/min for 3 days by a syringe pump (Harvard Apparatus, Holliston, MA). The culture medium contained 25 μM blebbistatin (ab120425, Abcam, Cambridge, MA) to ensure that the cells would not proliferate in the first part of the biological study [32]. A cytotoxicity kit (Fisher Scientific) was used to assess the cell viability within the transverse cross sections of hydrogels ( i.e.…”

Perivascular medical devices and drug delivery systems: Making the right choices

“…Similar observations were made when PCL or PLCL were employed for sirolimus and the paclitaxel preparation of perivascular DDS. [75,108,111,115,117]. A zero-order release duration of more than 21 days was achieved for paclitaxel and sirolimus, resulting in a 74 % reduction of IH [108].…”

“…This could result in a nonconformable covering of the graft and could also trigger injury to the vasa vasorum, which may in turn, induce IH formation [109]. To improve elasticity, PEG or methylated PEG was added as a plasticizer to improve the elastic properties of cuffs and wraps [11,76,110,111]. EVAc, usually used at 40 % vinyl acetate co-polymer composition, can form flexible matrices with ACCEPTED MANUSCRIPT satisfactory elastic properties [11,76,106,[112][113][114][115].…”