Leukocyte depletion during cardiopulmonary bypass has been demonstrated in animal experiments to improve pulmonary function. Conflicting results have been reported, however, with clinical depletion by arterial line filter of leukocytes at the beginning of cardiopulmonary bypass. In this study, we examined whether leukocyte depletion from the residual heart-lung machine blood at the end of cardiopulmonary bypass would improve lung function and reduce the postoperative inflammatory response. Thirty patients undergoing elective heart operations were randomly allocated to a leukocyte-depletion group or a control group. In the leukocyte-depletion group (n = 20), all residual blood (1.2 to 2.1 L) was filtered by leukocyte-removal filters and reinfused after cardiopulmonary bypass, whereas in the control group an identical amount of residual blood after cardiopulmonary bypass was reinfused without filtration (n = 10). Leukocyte depletion removed more than 97% of leukocytes from the retransfused blood (p < 0.01) and significantly reduced circulating leukocytes (p < 0.05) and granulocytes (p < 0.05) compared with the control group. Levels of the inflammatory mediator thromboxane B2 determined at the end of operation (p < 0.05) were significantly lower in the depletion group than in the control group, whereas no statistical differences in interleukin-6 levels were found between the two groups. After operation, pulmonary gas exchange function (arterial oxygen tension at a fraction of inspired oxygen of 0.4) was significantly higher in the leukocyte-depletion group 1 hour after arrival to the intensive care unit (p < 0.05) and after extubation (p < 0.05). There were no statistical differences between the two groups with respect to postoperative circulating platelet levels and blood loss, and no infections were observed during the whole period of hospitalization. These results suggest that leukocyte depletion of the residual heart-lung machine blood improves postoperative lung gas exchange function and is safe for patients who are expected to have a severe inflammatory response after heart operations.
Bovine pericardium has been extensively applied as the biomaterial for artificial heart valves and may potentially be used as a scaffold for tissue-engineered heart valves after decellularization. Although various methods of decellularization are currently available, it is unknown which method is the most ideal one for the decellularization for bovine pericardium. We compared three decellularization methods, namely, the detergent and enzyme extraction (DEE), the trypsin (TS), and the Triton X-100 and sodium-deoxycholate (TSD) method, to examine their efficacy on cell removal and their preservation of the mechanical function and the tissue matrix structure. Results indicated that decellularization was achieved by all the three methods as confirmed by hematoxylin-eosin staining, scanning electron microscopy, as well as quantitative DNA measurement. However, TS and TSD methods resulted in severe structural destruction of the bovine pericardium as shown by von Gieson staining and Gomori staining. Furthermore, both TS and TSD methods changed the mechanical property of the bovine pericardium, as evidenced by a lower elastic modulus, maximal-stress, maximal-disfiguration, maximal-load, and maximal-strain. In conclusion, the DEE method achieved both a complete decellularization and preservation of the mechanical function and tissue structure of the bovine pericardium. Thus, this method is superior to either the TS or the TSD method for preparing decellularized bovine pericardium scaffold for constructing tissue-engineered heart valves.
A biodegradable poly(ε-caprolactone) (PCL) was synthesized by ring-opening polymerization of ε-caprolactone catalyzed by Sn(Oct)2/BDO, followed by the heparin conjugation using EDC/NHS chemistry. The structure of the heparin-PCL conjugate was characterized by (1)H-NMR and GPC. The results of static contact angle and water uptake ratio measurements also confirmed the conjugation of heparin with the polyester. Its in vitro anticoagulation time was substantially extended, as evidenced by activated partial thromboplastin time (APTT) testing. Afterwards the conjugate was electrospun into small-diameter tubular scaffolds and loaded with Fibroblast Growth Factor 2 (FGF2) in aqueous solution. The loading efficiency was assayed by enzyme-linked immunosorbent assay (ELISA); the results indicated that the conjugate holds a higher loading efficiency than the blank polyester. The viability of released FGF2 was evaluated by MTT and cell adhesion tests. The amount and morphology of cells were significantly improved after FGF2 loading onto the electrospun heparin-PCL vascular scaffolds.
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