Tissue-engineered valved conduits in the pulmonary circulation.

OBJECTIVE: Bioprosthetic and mechanical valves and valved conduits are unable to grow, repair, or remodel. In an attempt to overcome these shortcomings, we have evaluated the feasibility of creating 3-leaflet, valved, pulmonary conduits from autologous ovine vascular cells and biodegradable polymers with tissue-engineering techniques. METHODS: Endothelial cells and vascular medial cells were harvested from ovine carotid arteries. Composite scaffolds of polyglycolic acid and polyhydroxyoctanoates were formed into a conduit, and 3 leaflets (polyhydroxyoctanoates) were sewn into the conduit. These constructs were seeded with autologous medial cells on 4 consecutive days and coated once with autologous endothelial cells. Thirty-one days (+/-3 days) after cell harvesting, 8 seeded and 1 unseeded control constructs were implanted to replace the pulmonary valve and main pulmonary artery on cardiopulmonary bypass. No postoperative anticoagulation was given. Valve function was assessed by means of echocardiography. The constructs were explanted after 1, 2, 4, 6, 8, 12, 16, and 24 weeks and evaluated macroscopically, histologically, and biochemically. RESULTS: Postoperative echocardiography of the seeded constructs demonstrated no thrombus formation with mild, nonprogressive, valvular regurgitation up to 24 weeks after implantation. Histologic examination showed organized and viable tissue without thrombus. Biochemical assays revealed increasing cellular and extracellular matrix contents. The unseeded construct developed thrombus formation on all 3 leaflets after 4 weeks. CONCLUSION: This experimental study showed that valved conduits constructed from autologous cells and biodegradable matrix can function in the pulmonary circulation. The progressive cellular and extracellular matrix formation indicates that the remodeling of the tissue-engineered structure continues for at least 6 months.

Tissue engineering of autologous aorta using a new biodegradable polymer.

BACKGROUND: Ovine pulmonary valve leaflets and pulmonary arteries have been tissue-engineered (TE) from autologous cells and biodegradable polyglycolic acid (PGA)-polyglactin copolymers. Use of this cell-polymer construct in the systemic circulation resulted in aneurysm formation. This study evaluates a TE vascular graft in the systemic circulation which is based on a new copolymer of PGA and polyhydroxyalkanoate (PHA). METHODS: Ovine carotid arteries were harvested, expanded in vitro, and seeded onto 7-mm diameter PHA-PGA tubular scaffolds. The autologous cell-polymer vascular constructs were used to replace 3-4 cm abdominal aortic segments in lambs (group TE, n = 7). In a control group (n = 4), aortic segments were replaced with acellular polymer tubes. Vascular patency was evaluated with echography. All control animals were sacrificed when the grafts became occluded. Animals in TE group were sacrificed at 10 days (n = 1), 3 (n = 3), and 5 months (n = 3). Explanted TE conduits were evaluated for collagen content, deoxyribonucleic acid (DNA) content, structural and ultrastructural examination, mechanical strength, and matrix metalloproteinase (MMP) activity. RESULTS: The 4 control conduits became occluded at 1, 2, 55, and 101 days. All TE grafts remained patent, and no aneurysms developed by the time of sacrifice. There was one mild stenosis at the anastomotic site after 5 months postoperatively. The percent collagen and DNA contents approached the native aorta over time (% collagen = 25.7%+/-3.4 [3 months] vs 99.6%+/-11.7 [5 months], p < 0.05; and % DNA = 30.8%+/-6.0 [3 months] vs 150.5%+/-16.9 [5 months], p < 0.05). Histology demonstrated elastic fibers in the medial layer and endothelial specific von Willebrand factor on the luminal surface. The mechanical strain-stress curve of the TE aorta approached that of the native vessel. A 66 kDa MMP-2 was found in the TE and native aorta but not in control group. CONCLUSIONS: Autologous aortic grafts with biological characteristics resembling the native aorta can be created using TE approach. This may allow the development of "live" vascular grafts.

Automated face analysis by feature point tracking has high concurrent validity with manual FACS coding.

The face is a rich source of information about human behavior. Available methods for coding facial displays, however, are human-observer dependent, labor intensive, and difficult to standardize. To enable rigorous and efficient quantitative measurement of facial displays, we have developed an automated method of facial display analysis. In this report, we compare the results with this automated system with those of manual FACS (Facial Action Coding System, Ekman & Friesen, 1978a) coding. One hundred university students were videotaped while performing a series of facial displays. The image sequences were coded from videotape by certified FACS coders. Fifteen action units and action unit combinations that occurred a minimum of 25 times were selected for automated analysis. Facial features were automatically tracked in digitized image sequences using a hierarchical algorithm for estimating optical flow. The measurements were normalized for variation in position, orientation, and scale. The image sequences were randomly divided into a training set and a cross-validation set, and discriminant function analyses were conducted on the feature point measurements. In the training set, average agreement with manual FACS coding was 92% or higher for action units in the brow, eye, and mouth regions. In the cross-validation set, average agreement was 91%, 88%, and 81% for action units in the brow, eye, and mouth regions, respectively. Automated face analysis by feature point tracking demonstrated high concurrent validity with manual FACS coding.