Construction of an acellular porcine aortic valve / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare a porcine aortic valve (PAV) free of the cellular components.</p><p><b>METHODS</b>The cellular components of porcine PAV were completely removed using trypsin and Triton X-100, and the acellular PAV was examined microscopically with HE staining with its physical and chemical properties assessed. Transmission electron microscopy was used to observe the integrity of the collagen and elastin and the DNA contents in the PAV was detected to confirm the total removal of the cellular components. With the fresh PAV as the control, small pieces of the acellular PAV were implanted into the subcutaneous tissues of 4 rabbits, and 4 weeks after the implantation, the implants were harvested for microscopic observation.</p><p><b>RESULTS</b>The cellular components were effectively removed from the cusps and roots of the PAV by trypsin and TritonX-100, with marked soluble protein loss [(0.24-/+0.04)% vs (0.48-/+0.12)%] and significantly increased water content [(92.2-/+1.5)% vs (89.2-/+1.6)%]. The acellular PAV still maintained good fibrous scaffold structure and the shrinkage temperature and tension at fracture underwent no significantly changes [(67.9-/+1.0) degrees celsius; vs (68.8-/+0.8) degrees celsius; and (489.3-/+19.0) g/mm2 vs (540.7-/+19.5) g/mm2, respectively]. The PAVs implanted in rabbits showed only mild tissue reaction with a few infiltrating neutrophils, lymphocytes and plasmocytes observed 4 weeks later. The accelular PAV caused obviously milder inflammatory reactions than fresh PAV.</p><p><b>CONCLUSIONS</b>The acellular PAV prepared by treatment with trypsin and Triton X-100 retains good fibrous scaffold structure and mechanical strength with low antigenicity.</p>

Effects of intensive insulin therapy on plasma nitric oxide and endothelin-1 levels in patients undergoing cardiac surgery under cardiopulmonary bypass / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the effects of intensive insulin therapy on plasma nitric oxide (NO) and endothelin-1 (ET-1) levels in patients undergoing cardiac valve replacement under cardiopulmonary bypass (CPB).</p><p><b>METHODS</b>A total of 36 patients were randomly assigned to routine therapy (RT) group and intensive insulin therapy (IT) group, with 18 patients in each group. The blood glucose levels during surgery were maintained at 3.9 to 10.0 mmol/L and those after surgery at 3.9 to 6.1 mmol/L in IT group, whereas patients in RT group didn't undergo the treatment of controlling glucose levels during operation and maintained below 13.9 mmoVL after operation. Levels of plasma NO and ET-1 in both groups were respectively measured before surgical anesthesia, at the initiation of CPB, and 0 h, 4 h, 12 h, 24 h and 48 h after the termination of CPB.</p><p><b>RESULTS</b>In RT group, plasma NO concentration was decreased since the initiation of CPB [from (68.2 +/- 16.3) micromol/L to (67.8 +/- 8.4) micromol/L] and reached the trough at the termination of CPB [ (60.0 +/- 10.2) micromol/L, P < 0.05 compared with that before anesthesia]. Then it began to increase and neared to the preoperational level 48 h after the termination of CPB. In contrast, plasma ET-1 concentration was increased since the initiation of CPB [from (62.2 +/- 10.2) ng/L to (68.3 +/- 10.8) ng/L] and reached the peak at the termination of CPB [ (112.5 +/- 18.6) ng/L, P < 0.01 compared with that before anesthesia]. Then it began to decrease and reached the preoperational level 24 h after the termination of CPB. In IT group, however, the changes of NO and ET-1 levels at different time points during CPB and thereafter didn't reach the significance as compared with those before anesthesia.</p><p><b>CONCLUSIONS</b>Intensive insulin therapy may relieve the changes of CPB-induced NO and ET-1 levels during cardiovascular surgery, which suggests its protective effects on cardiovascular function.</p>

Experimental study of differentiation of canine bone marrow mesenchymal stem cell into fibroblasts in vitro / 中华外科杂志

<p><b>OBJECTIVE</b>To explore the way of stably inducing canine bone marrow mesenchymal stem cells (BMSCs) to differentiate into fibroblasts and myofibroblasts in vitro, and provide seed cells for fabricating tissue engineering heart valves (TEHV).</p><p><b>METHODS</b>Adult canine BMSCs were separated by a gradient centrifugation on Percoll (density 1.073 g/ml), then the cells were incubated in low-glucose Dulbecco Eagle's minimum essential medium (LG-DMEM) with 10% bovine calf serum. Cell phenotype were identified by immunohistochemistry staining. The second and third generation of BMSCs were committedly induced by conditioning culture medium, which were detected by immunohistochemistry staining. The induced-BMSCs were freezed, preserved and resuscitated after 7 d to observe the cell growth, proliferation and function.</p><p><b>RESULTS</b>BMSCs deriving from the bone marrow mononuclear cells separated by a Percoll gradient were positive expression of alpha-smooth muscle antibody, vimentin and negative expression of CD34, laminin. About (50 +/- 3)% induced-BMSCs were positive expression of laminin. Approximately (85 +/- 3)% freezed induced-BMSCs could be resuscitated. And the growth, proliferation and function were well.</p><p><b>CONCLUSION</b>BMSCs could be committedly induced to differentiate into fibroblasts and myofibroblasts in vitro. It is suitable to be the seed cells.</p>

Construction of a tissue-engineered valve with decellular porcine aortic valve scaffold in the abdominal aorta of canine / 中华外科杂志

<p><b>OBJECTIVE</b>To explore an experimental method for construction of tissue-engineered heart valve (TEHV) in canine abdominal aorta.</p><p><b>METHODS</b>The decellular porcine aortic valve (PAV) leaflets seeded with canine vessel interstitial cells and endothelial cells (ECs) were implanted into 6 canine abdominal aortas. Valve specimens were obtained respectively at the end of 4, 6, 8 and 10 weeks after implantation were studied for morphology, histology and immunohistochemistry.</p><p><b>RESULTS</b>(1) After 4 weeks implantation, multiple layers of cells grew into peripheral portion of valve scaffold, while new extracellular matrix appeared, and original scaffold tissue was partially absorbed. (2) At the end of 10th week after implantation, the decellular PAV scaffold disappeared completely and was substituted by recipient cells and new extracellular matrix. The interstitial cells in matrix was mainly consisted of fibroblasts and myofibroblast. The matrix was mainly composed by type I, III collagen, some elastic fibers with neutral and acid mucopolysaccharide. (3) Surface of valve leaflets were covered with endothelial cells.</p><p><b>CONCLUSIONS</b>(1) TEHV is primarily constructed with recellularized PAV after implantation into canine abdominal aorta for 10 weeks. (2) Heterotopic implantation into the abdominal aorta is an alternative experimental procedure to study the TEHV.</p>