RESUMO
BACKGROUND: Revascularization is a challenge for the tissue-engineered bone carrying cells after implanted into human body. Previous studies have found that tanshinol can improve the functions of endothelial progenitor cells and exert vascular protective effects.OBJECTIVE: To prepare the β-calcium phosphate (β-TCP) scaffold with tanshinol coating, and to observe its cytocompatibility.METHODS: The β-TCP scaffolds coated with 10-7, 10-6 and 10-5 mol of tanshinol were constructed by negative pressure absorption method. The distribution of tanshinol coating on the scaffold was observed using scanning electron microscopy,and the inner ingredients were analyzed by infrared spectrum. Human endothelial progenitor cells (hEPCs) were cultured in the extracts of β-TCP and β-TCP scaffolds with 10-7, 10-6, 10-5 mol of tanshinol coatings, respectively. The cell proliferation was detected at 2, 4, 6, 8 and 10 days of culture; the levels of nitric oxide and vascular endothelial growth factor in the supernatants were detected at 1, 7 and 14 days of culture; the lumen formation on the matrigel was observed after 14-day culture. hEPCs were respectively seeded onto the β-TCP and β-TCP scaffolds with different dosages of tanshinol coating,and then the cell growth was observed under scanning electron microscope at 7 days.RESULTS AND CONCLUSION: The tanshinol coating evenly distributed on the inner surface of the pores, and its crystalline structure became dense with dosage increasing. Infrared spectrum analysis revealed no changes in the characteristic absorption peak of tanshinol and TCP in the scaffold. The β-TCP scaffolds with tanshinol coating could promote the proliferation of hEPCs, especially the scaffolds with 10-6 and 10-5 mol tanshinol coating. Compared with the β-TCP scaffold, the scaffolds with 10-6 and 10-5 mol tanshinol coating significantly upregulated the nitric oxide level at 14 days of culture, and significantly increased the level of vascular endothelial growth factor at 7 and 14 days of culture (P <0.05). Although it could be found in all β-TCP scaffolds with tanshinol coating, the lumen formation was the maturest in the scaffold with 10-5 mol tanshinol coating. These results suggest the β-TCP scaffolds with tanshinol coating can promote the proliferation and endothelial differentiation of hEPCs, and hold a good cytocompatibility.
RESUMO
BACKGROUND: There are various methods to observe and detect early angiogenesis in the process of entochondrostosis, but each holds certain deficiencies.OBJECTIVE: To explore the possibility of tetracycline and alizarin complexone as an indirect marker ofangiogenesis in the process of entochondrostosis.METHODS: New Zealand white rabbit models of bilateral radial bone defects were prepared, followed by β-tricalcium phosphate implantation, and then given the injection of tetracycline and alizarin complexone at 1 and 15 days,respectively. Samples were collected at 28 days, some of which were observed using fluorescence/light microscope after ink perfusion and hard tissue slicing, and the others were decalcified and observed using immunohistochemistry. The uniformity between lumen structures labeled with bone affinity fluorescein and vascular structures marked by immunohistochemistry and ink perfusion was compared.RESULTS AND CONCLUSION: The lumen structure labeled with bone affinity fluorescein was confirmed to be a CD34 positive vascular structure. Under the fluorescence microscope, the bone affinity fluorescein labeled vascular morphology was consistent with ink perfusion-labeled, and black ink lines could be observed in the lumen structures labeled with bone affinity fluorescein after ink perfusion. In addition, the color of the lumen labeled with fluorescein was more gorgeous,three-dimensional structure more vivid, and the vascular evolution process distinguished more easily by different fluorescein colors, exhibiting unique advantages. Therefore, it is available to detect the early angiogenesis in the process of entochondrostosis.
RESUMO
Objective It is still controversial how to deal with the congenital heart disease with advanced pulmonary hypertension.The choices of treatment for these patients must depend on the character and degree of pulmonary vascular change.The pulmonary vascular change correlates well with hemodynamics and acute pulmonary vasoreactivity test.Therefore,it will play an important role to obtain accurately these evaluations.Methods This paper studies 85 cases (38 males and 46 females) with congenital heart disease complic ated with advanced pulmonary hypertension and bidirectional shunt,which are diagnosed by echocardiography.Whose age range is (22.8 ± 16.5 ) and weight range (46.4 ± 12.1 ) kg.Among those cases,42 are VSD,11 are ASD,9 are VSD with patent ductus arteriosu,7 are patent ductus arteriosu,5 are ASD with VSD,and 11 are the others.With congenital heart disease complicated with advanced pulmonary hypertension and bidirectional shunt,which are diagnosed by echocardiography.There were floating catheter retention of 4 to 13 days ( average 7.5 days).Through the right subclavian vein or jugular vein,floating catheter is inserted into pulmonary artery.Pulmonary artery pressure is recorded continuously and analyzed.Hemodynamics,acute pulmonary vasoreactivity test and drug sensitivity test are evaluated.Differences of the hemodynamics are analyzed between echocardiography and floating catheter examination.Results Compared with floating catheter examination,there are not significantly difference for the diagnosis of advanced pulmonary hypertension(98.8％ vs.100％,P ＞ 0.05),less accurately diagnosis (64.3％ vs.100％,P ＜ 0.05 ) and higher (H)false positive rate (8.4％ vs.0,P ＜ 0.05) for Eisenmenger syndrome in echocardiography examination.Floating catheter examination shows that mean systolic pulmonary artery pressure(PAP)/mean systolie body artery blood pressure (BP) and mean PAP / mean BP were equal to 1.22 ± 0.35 and 1.07 ± 0.11 respectively.The patients presented total pulmonary vascular resistance of ( 17.6 ± 8.3) Wood units.Bidirectional shunt volume is more than 30％ cardiac outputs in 7 cases.Apart from 14 cases with Eisenmenger syndrome,total pulmonary vascular resistance is ( 11.3 ± 3.7) Wood units.Acute pulmonary vasoreactivity test presents positive in 67 cases.After drug sensitivity test,14 cases with Eisenmenger syndrome underwent medical treatment and waited lung transplantation or heart-lung transplantation.Floating catheter examination happened hematoma(2/85 ) in 2 patients.71 cases underwent surgical treatment.Operative complications included right heart failure (6/71) and pulmonary infection (5/71).Operative mortality was 5.6％ (4/71).The main causes of death were right heart failure.Compared with preoperative arterial oxygen saturation,postoperative arterial oxygen saturation increased by (9.7 ± 4.1 ) ％ ( P ＜ 0.05 ).Conclusion Floating catheter examination can evaluate accurately pulmonary hypertension,hemodynamics,acute pulmonary vasoreactivity test,drug sensitivity test and bidirectional shunt volume.All the patients,who were diagnosed as advanced pulmonary hypertension with bidirectional shunt by echocardiography,should undergo floating catheterization.Floating catheter examination can provide reliable,objective theoretical basis for the choice of treatment in the congenital heart diseases with advanced pulmonary hypertension.
RESUMO
To study the effects of different pH HEPES-KH reperfusate solution on immature myocardial protection, isolated perfused Langendorff model from immature rabbit hearts were developed formed. Control group (C) was perfused only with pH 7.4 HEPES-KH solution for 90 min. Ischemia/reperfusion group (group I/R) was perfused with pH 7.4 HEPES-KH solution before ischemia or after ischemia. Experimental group (group E), after ischemia, was perfused with pH 6.8, pH 7.1 and pH 7.4 HEPES-KH solutions for 5 min, 5 min, and 20 min, respectively. The left ventricular function recovery, MWC, LDH and CK leakage, MDA, ATP content, and SOD activity were determined. Our results showed that the left ventricular function recovery, ATP content and SOD activity in group E were higher than those of group I/R (P < 0.05). MWC, MDA content, LDH and CK leakage in group E were lower than those of group I/R (P < 0.05). These findings suggested that pH paradox might be one of important mechanisms for immature myocardial ischemia-reperfusion injury, and acidic perfusate, at the beginning of reperfusion, might attenuate pH paradox and ameliorate functional recovery in isolated perfused immature rabbit hearts.
Assuntos
Miocárdio/metabolismo , Distribuição Aleatória , Superóxido Dismutase/metabolismoRESUMO
To study the effects of different pH HEPES-KH reperfusate solution on immature myocardial protection, isolated perfused Langendorff model from immature rabbit hearts were developed formed. Control group (C) was perfused only with pH 7.4 HEPES-KH solution for 90 min. Ischemia/reperfusion group (group I/R) was perfused with pH 7.4 HEPES-KH solution before ischemia or after ischemia. Experimental group (group E), after ischemia, was perfused with pH 6.8, pH 7.1 and pH 7.4 HEPES-KH solutions for 5 min, 5 min, and 20 min, respectively. The left ventricular function recovery, MWC, LDH and CK leakage, MDA, ATP content, and SOD activity were determined. Our results showed that the left ventricular function recovery, ATP content and SOD activity in group E were higher than those of group I/R (P < 0.05). MWC, MDA content, LDH and CK leakage in group E were lower than those of group I/R (P < 0.05). These findings suggested that pH paradox might be one of important mechanisms for immature myocardial ischemia-reperfusion injury, and acidic perfusate, at the beginning of reperfusion, might attenuate pH paradox and ameliorate functional recovery in isolated perfused immature rabbit hearts.
Assuntos
Animais , Feminino , Masculino , Coelhos , Soluções Cardioplégicas , Farmacologia , Coração , Parada Cardíaca Induzida , Concentração de Íons de Hidrogênio , Isquemia Miocárdica , Metabolismo , Traumatismo por Reperfusão Miocárdica , Metabolismo , Miocárdio , Metabolismo , Distribuição Aleatória , Superóxido Dismutase , MetabolismoRESUMO
Ninety fresh umbilical cords of newborns were harvested and dissected. There were 86 cords with double arteries and single vein and 4 cords with only 1 artery and 1 vein. The outer and inner perimeter of single artery, double arteries and the vein was 3 8?0 3mm vs 6 2?0 3mm, 2 4?0 2mm vs 4 6?0 2mm and 10 0?0 4mm vs 10 8?0 4mm at empty and flatened state, respectively. The diameter of vein at 20mmHg filling pressure and single and double arteries at 80mmHg was 7 2?0 3mm, 4 8?0 1mm and 3 9?0 8mm, respectively. According to these values, various drainage and perfusion metal tubes were designed and evaluated both in vitro and in vivo. By using these tubes,extracorporeal circulation was established successfully via umbilical artery and vein in a newborn who suffered from cardiac arrest after birth. The results suggest that CPB through umbilical artery and vein is simple, practical and effective.