Figure 1. Schematic of EpîNanoPaint and its application by pen-brushing.
A. Illustration of ahNP structure. The ahNP has a hydrophobic PCL core and is adhesive because of the sulfo-NHS groups on the PEG arms. For ahNP immobilization on extracellular matrix (ECM), sulfo-NHS reacts with NH2 on ECM proteins to form an amide bond. The sulfo-NHS byproduct is highly water-soluble and can be easily removed.
B. Size and zeta potential of PAMAM-PCL-OH and ahNP from DLS. Quantification: Data are presented as mean ± SEM, n= 3 independent repeat experiments.
C. TEM image demonstrating the spherical shape of ahNP.
D. Cumulative release of pinometostat from ahNP at 37°C for 100 days. Quantification: Data are presented as mean ± SEM, n=3 independent repeat experiments.
E. SEM image of human adventitial ECM33. Scale bar: 1 μm.
F. The sulfo-NHS (N-hydroxysulfosuccinimide) ester on ahNP forms an amide bond with NH2 rich on the adventitial ECM.
G. Schematic of epi-vascular application by pen-brushing ahNP onto the vessel adventitia.
H. Schematic of ahNP immobilization on the adventitial ECM.
I. Confocal image (collected 7 days after brushing) showing adventitial Cy5 signal and medial (M) elastin auto-fluorescence (scale bar: 50 μm).
J. Effect of Pino/ahNP on H3K79me2. IH was induced by open-surgery and balloon injury in obese Zucker rat common carotid arteries, and Pino-loaded ahNP or empty ahNP was pen-brushed on the adventitia of the injured arteries. Arteries were collected 28 days later and cross-sections were used for immunofluorescence staining of H3K79me2. Neointima is demarcated by arrow heads. Scale bar: 50 μm.
K. Quantification for J. Shown are enlarged merge images. Scale bar: 50 μm. The values from 4–6 sections were pooled to generate the mean for each animal. The means from 4 animals in each group were then averaged to produce the final mean ± SEM (n = 4 rats). Unpaired Student t-test: **p<0.01