Clearance of beta-amyloid is facilitated by apolipoprotein E and circulating high-density lipoproteins in bioengineered human vessels
- University of British Columbia, Canada
Figures
Figure 1
Schematic representation of bioengineered vessels.
Scaffolding material was prepared into a tubular shape approximately 2 mm in diameter and 15 mm long (a). Scaffolds were sequentially seeded with primary human umbilical vein smooth muscle cells …
Figure 2 with 1 supplement
Histological structure of bipartite bioengineered vessels.
(a) Haematoxylin and Eosin staining revealed dense tissue formation composed of cells and extracellular matrix (ECM). (b) Picrosirius staining confirmed secretion of collagen. Further immunostaining …
Figure 2—figure supplement 1
Aβ40 and Aβ42 accumulate similarly in tissues seeded with cells originating from umbilical cord or cortex.
Aβ40 and Aβ42 monomers (1 μM) were injected into the tissue chamber (antelumen) of tissues seeded with SMC from umbilical cords or cerebral arteries and incubated for 24 hr under flow conditions …
Figure 3 with 1 supplement
Aβ40 and Aβ42 accumulation within and transport through bipartite vessels.
Aβ40 and Aβ42 monomers (0, 0.1, 1.0 and 10 μM) were injected into the tissue chamber (antelumen) and incubated for 48 hr under flow conditions (a–f). Aβ deposition within bioengineered vessels was …
Figure 3—figure supplement 1
Histological structure of Aβ deposited in bioengineered vessels.
(a) Confocal immunostaining against OC fibrils and α-SMA confirmed the deposition of fibrillized Aβ40 and Aβ42 outside of the cells. (b) Confocal immunostaining against 6E10 and Col-IV confirmed the …
Figure 4
Lipoproteins reduce Aβ42 accumulation within bioengineered bipartite vessels.
Aβ40 and Aβ42 monomers (1 μM) were incubated without or with recombinant apoE (ratio 25:1) for 2 hr at 37°C before injection into the anteluminal tissue chamber. The levels of transported Aβ was …
Figure 5
HDL facilitates Aβ42 transport and reduces accumulation in bioengineered bipartite vessels in the presence of recombinant apoE.
Aβ40 and Aβ42 monomers (1 μM) were incubated without or with recombinant apoE2 (a–d) or apoE4 (e–h) (ratio 25:1) for 2 hr at 37°C before injecting into the tissue chamber in the absence or presence …
Figure 6 with 3 supplements
Histological structure of tripartite bioengineered vessels.
Immunostaining against CD31 confirmed the presence of an EC monolayer on the luminal side of tripartite vessels (a), the expression of α-SMA confirmed the smooth muscle phenotype of cells in the …
Figure 6—figure supplement 1
Astrocytes in bioengineered tissues express aquaporin four and NDRG2.
The astrocytes in tripartite tissue were further analysed with immunostaining against aquaporin 4 (AQ4) (a) or NDRG2 (b) with astrocytic markers (GFAP, S100β). Bars represent 50 μm.
Figure 6—figure supplement 2
HUVEC express the brain Glut-1 transporter when culture in bioengineered vessels.
The EC bioengineered tissues were further analysed with immunostaining against Glut-1 and the EC marker CD31 in comparison with human brain and umbilical cord. Bars represent 200 μm.
Figure 6—figure supplement 3
Endothelia of bioengineered vessels are impermeable to FITC-dextran.
Barrier integrity was assessed by measuring permeability of 250 μg/ml of 4 kDa or 40 kDa FITC-dextran circulated through the lumen for 1 hr. Graphs represent mean ±SEM for at least three independent …
Figure 7
HDL facilitates Aβ transport and reduces accumulation in bioengineered tripartite vessels expressing native astrocyte apoE.
(a–d) Tripartite vessels were treated with the LXR agonist GW3965 (0.8 μM) for 72 hr to stimulate astrocyte apoE3 secretion. Aβ40 and Aβ42 monomers (1 μM) were injected in the tissue chamber in the …
Figure 8
Amyloid fibrils are transported more slowly but accumulate similarly between bipartite and tripartite tissues.
(a–d) Pre-aggregated or monomeric Aβ40 and Aβ42 (1 μM) were injected in the tissue chamber. The levels of transported Aβ were measured by ELISA from samples collected from the luminal circulating …
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- https://doi.org/10.7554/eLife.29595.016
