Figure 1.
The newly developed integrated vascular bioreactor system.
A, (1) the intelligent bionic culture setting; (2) the culture biochamber; (3) silicone tube; (4) the stepper motor-driven pump; (5) the gas/liquid and liquid/liquid exchanger. B, the high-magnification image of the culture chamber. C, the DO sensor and PH sensor. D, the DO controller and PH controller. E, the flow control valve to control the carbon dioxide, nitrogen, and oxygen fluid. F, the force monitor.
Figure 2.
The schema of the perfusion-based bioreactor systems.
(1) air inlet; (2) air outlet; (3) fluid inlet; (4) fluid outlet; (5) the gas/liquid and liquid/liquid exchanger; (6) silicone tube; (7) the stepper motor-driven pump; (8) the culture biochamber; (9) the tissue-engineered vascular graft; Arrow, the liquid flow; (10) the pressure sensor.
Figure 3.
The real-time pressure and pulse rates generated by the motor-driven pump monitored by a baroceptor.
Figure 4.
A, The non-computer-controlled culture system: (1) the linear motor-driven pump; (2) the culture chamber; (3) the instrument control system.
Figure 5.
The phenotypes of the newly developed small-diameter vessel cultured for 2 weeks.
A, the gross appearance of the newly developed vessel. B, HE staining analysis of histologic sections of the newly developed vessels. C and D, SEM analysis of endothelial cells and smooth muscle cells, respectively. E, the comparison of the survival of SMCs cultured under dynamic conditions with that of SMCs cultured under static conditions determined by MTT assay after 2-week culture. Bars are mean ± S.D. from three independent experiments. *Significantly different from static conditions, P<0.05.
Figure 6.
Determination of endothelin-1, NO and PGI2 secretion of vessels.
A, immunohistochemistry for endothelin-1 expression of the newly developed small-diameter vessel. Bars are mean ± S.D. from three independent experiments. *Significantly different from static conditions, P<0.05. B, total NO secretion determined by use of a chemiluminescence NO detector. Bars are mean ± S.D. from six independent experiments. *P<0.05. C, total PGI2 secretion determined as its metabolite 6-keto-PGF1α assayed by ELISA. Bars are mean ± S.D. from three independent experiments. *P<0.05. D, the morphology of platelet adhesion on the inner lumen surface of tissue engineered vessels was examined by SEM.
Figure 7.
Biochemical assay of newly developed grafts.
A and B immunohistochemistry for calponin and smooth muscle α-actin, respectively. C, elastica van Gieson staining of elastin. D, determination of collagen deposition of newly developed grafts by Masson trichrome staining. Bars are mean ± S.D. from three independent experiments. *P<0.05.
Figure 8.
qRT-PCR analysis of collagen-related gene (Collagen type I, III and IV) expression in cells cultured static conditions and dynamic conditions at indicated times.
Bars are mean ± S.D. from three independent experiments. *P<0.05.
Figure 9.
The mechanical properties of the constructs and normal rat vessels as measured using biaxial tensile testing of ring sections of the constructs.
Bars are mean ± S.D. from four-seven independent experiments. *Significantly different from static condition group, P<0.05.
Figure 10.
The comparison of cell survival and morphous of constructs between cultured in the computer-controlled bioreactor and the non-computer-controlled bioreactor.
A, SEM analysis of endothelial cells and smooth muscle cells cultured both in the computer-controlled and non-computer-controlled system after 2 weeks, respectively. B, the proliferation of endothelial cells and smooth muscle cells cultured both in the computer-controlled and non-computer-controlled system after 2 weeks determined by MTT assay. Bars are mean ± S.D. from three independent experiments. *P<0.05.