Figure 7.

(7A) Proliferation of EaRASMCs is not impacted by coculture with DOX-SMPs (cathepsin K Ab-conjugated or unmodified), although cell proliferation in the cultures treated with the Ab-conjugated SMPs was significantly lower than that cultured with the unmodified SMPs. DOX loading within the SMPs was 2% w/w. SMP concentration in the cultures was 0.2 mg/ml. Cells were harvested at 1 and 21 days after initial seeding (mean ± SD; n = 3 cultures per group; # denotes significance of differences versus unconjugated DOX SMPs, deemed for p< 0.05). (7B) Cathepsin K antibody conjugation of DOX-SMPs does not alter their pro-elastogenic effects on cultured EaRASMCs. SMP-untreated EaRASMC cultures were investigated as treatment controls. All cell layers were cultured for 21 days. Amounts of deposited elastic matrix, comprised of both the alkali-soluble and alkali-insoluble elastin fractions were normalized to DNA content of the respective cell layers (mean ± SD; n =3 cultures per group; # denotes significance of differences relative to treatment controls deemed for p < 0.05). (7C) Transmission electron micrographs showing effects of cathepsin K-Ab-conjugated and unconjugated DOX-SMPs on elastic matrix deposition in TNF-α activated SMC cultures. Elastic matrix deposition was sparse in the SMP-untreated cultures and few amorphous elastin deposits and no mature fibers were seen (1). Numerous forming elastic fibers were seen in the SMP-treated cultures (2), with a greater number of amorphous elastin deposits (white arrows) associated with the microfibrillar components in the cultures that received the cathepsin-K Ab-modified DOX-SMPs (3). Scale bars: 1 μm.