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In this report, the feasibility of producing a completely biological TEBV exclusively …


Biology Articles » Bioengineering » A completely biological tissue-engineered human blood vessel » Figures

Figures
- A completely biological tissue-engineered human blood vessel

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Figure 1. Organization of the TEBV. A) Macroscopic view of a mature TEBV (9 wk of adventitial maturation). The vessel is self-supporting when removed from culture medium (open lumen = 3 mm). Note that the various layers now form a continuous vascular wall (inset, graduation = 1 mm). B) Paraffin cross section of the vascular wall stained with Masson's trichrome shows collagen in blue-green and cells in dark purple. Aside from an oversized internal elastic lamina (IM=125 µm), the histology is similar to that of a muscular artery with a large media (M=320 µm) and a surrounding adventitia (A=235 µm). C) En face view of the endothelium seeded on the IM. Cytoplasmic green fluorescence reveals cell viability, metabolic activity, and degree of confluence. Red fluorescence (DiI-ac-LDL uptake) confirms cell viability and the endothelial nature of the cells. Blue fluorescence shows the characteristic von Willebrand factor expression in ECs (orange = red + green; pink = red + green + blue). Scale bar = 25 µm. D) Frozen cross section of the media–adventitia junction (arrows) stained for desmin (nuclei are stained blue). Scale bar = 50 µm. E) Frozen cross section of the adventitia double stained for elastin (green) and vimentin (red). Scale bar = 50 µm.

Figure 1

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Figure 2. Mechanical strength of the TEBV. A) Wall thickness of adventitia as a function of culture time before sheet detachment and rolling (maturation period is 5 wk in all cases; *significantly different from 4 wk (P*significantly different from 5 wk (P*significantly different from 5 wk (P*P

Figure2

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Figure 3. Extracellular matrix ultrastructure and collagenase expression. A) Transmission electron micrograph of the adventitial matrix. Uranyl acetate and lead citrate stain. Scale bar = 500 nm. B) Gelatin zymogram showing gelatinase activity in conditioned culture medium. Lanes 1 and 2: fibroblast sheet prior to rolling (FIB) and 48 h after rolling (adventitia = ADV). Lanes 3 and 4: SMC sheet prior to rolling (SMC) and 48 h after rolling on a IM (vascular media = VM). Identical results were obtained without a IM. Lanes 5 to 8: addition of the adventitia over the VM. Note that albumin bands (dark 68 kDa band) are lighter in conditioned mediums from sheets prior to rolling because different sample volumes were used to take into account the total volume of culture medium.

Figure 3

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Figure 4. Inhibition of platelet adhesion by the endothelium. Scanning electron micrographs of unendothelialized IM (A) promoted platelet adhesion and activation whereas endothelialized IM (B) almost completely inhibited the process.

Figure 4

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Figure 5. In vivo grafting of the TEBV. A) Angiogram of the lower limbs 7 days after implantation. Two patent TEBV are visible (arrows) providing normal blood flow in both legs. B) After angiographic examination, the TEBV was explanted along with adjacent segments of the femoral artery, gently flushed with saline, fixed in formaldehyde, longitudinally slit open, and pinned down to expose the luminal surface. Blood infiltrations are seen inside the vessel wall. The luminal surface is free of thrombus. Anastomosis showed no signs of deterioration. Graduation = 1 mm.

Figure 5

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