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1.
目的 探讨重组骨生长肽对组织工程性骨肌皮瓣的影响。方法 自2004年12月至2005年5月,采用新西兰大耳白兔36只,在其髂后上棘抽取骨髓基质干细胞进行体外培养,诱导分化为成骨细胞。然后将第3代细胞与羟基磷灰石/磷酸三钙(HA/TCP)、重组骨生长肽结合,共同培养14d后,经相差显微镜、扫描电镜观察体外细胞的生长情况,最后分别将72块复合物植入36只白兔的双侧背阔肌中,构成组织工程骨肌皮瓣。含重组骨生长肽的一侧为实验组;含成骨细胞的一侧为对照组。术后6周,进行大体解剖观察,行HE染色,观察骨肌皮瓣的生长情况。结果 骨髓基质干细胞可以诱导分化为成骨细胞,钙盐沉积,可形成不透光的矿化结节。在不同时间的骨髓基质干细胞,实验组ALP活性较对照组高(P〈0.05),并表现为时间依赖性。扫描电镜显示:支架材料表面和孔隙内均有成骨细胞生长,有良好的增殖活动。两组均可形成肌骨瓣,但实验组在成骨速度、成骨质量、血管化程度等方面均优于对照组。结论 应用重组骨生长肽可成功地构建组织工程性骨肌皮瓣。  相似文献   

2.
目的:观察大鼠骨髓基质干细胞(MSCs)与经胶原涂层处理的煅烧骨(CCB)复合行体外培养时贴附、增殖能力的变化,探讨构建组织工程骨体外培养的最适时间。方法:制备鼠尾胶原(I型胶原),无菌条件下,将煅烧骨置于胶原液中,2h后取出骨块,置滤网中将孔内外的胶原溶液沥干。体外培养的大鼠MSCs经矿化液诱导向成骨细胞分化。将分化后的成骨细胞滴加到涂胶原的煅烧骨块上,同时以未涂胶原的煅烧骨块为对照组。3天,7天时取材,扫描电镜下观察实验组与对照组成骨细胞在煅烧骨表面的贴附数量及形态,用细胞计数法测定细胞在两组材料的生长曲线,组织化学方法检测成骨细胞的ALP活性。结果:扫描电镜发现实验组成骨细胞的贴附数量显著高于对照组(P〈0.01)。细胞计数认为实验组细胞在材料上粘附数量多,增殖速度快。实验组ALP活性明显高于对照组(P〈0.01)。结论:胶原修饰的煅烧骨具有良好的组织相容性,能明显提高成骨细胞的贴附,并且促进成骨细胞活性维持,体外培养7~8天时,材料上的细胞数达到最大,应尽快植入体内。  相似文献   

3.
成骨生长肽对大鼠骨髓基质细胞增殖和成骨分化的作用   总被引:2,自引:0,他引:2  
肖毅  王建国  白增亮 《中国骨伤》2008,21(11):843-845
目的:观察成骨生长肽诱导体外培养的大鼠骨髓基质细胞增殖及向成骨分化的作用。方法:取6周龄SD大鼠,贴壁法分离培养骨髓基质细胞,在不同浓度成骨生长肽的诱导下,观察细胞形态变化,绘制骨髓基质细胞生长曲线,碱性磷酸酶和钙结节组织化学染色。结果:成骨生长肽对骨髓基质细胞增殖和成骨分化的作用呈剂量依赖性:成骨生长肽浓度在10^-10及10^-11mol/L时促进骨髓基质细胞增殖,而在10^-8及10^-9mol/L时对骨髓基质细胞的增殖略有抑制作用;成骨生长肽浓度在10^-10及10^-11mol/L时骨髓基质细胞碱性磷酸酶染色基本呈阴性,与对照组相比差异无统计学意义,而在10^-8及10^-9mol/L浓度下可以显著提高骨髓基质细胞碱性磷酸酶染色的阳性率。结论:成骨生长肽可以明显促进大鼠骨髓基质细胞增殖及向成骨细胞分化,其促成骨活性具有显著的浓度依赖性。  相似文献   

4.
目的探讨利用组织工程方法,以小肠粘膜下层为支架材料复合成骨诱导后的骨髓基质干细胞构建骨组织的可行性。方法将取自兔骨髓中的骨髓基质干细胞经成骨诱导液诱导后,与经处理的猪小肠粘膜下层在体外共培养。1周后,将共培养的猪小肠粘膜下层埋置于无胸腺裸鼠皮下。分别在不同时间进行扫描电镜、透射电镜、组织学和免疫组织化学观察。结果体外培养时,见细胞与材料粘附良好,且分泌大量的细胞外基质,细胞分化、增殖活跃。大体观察植入体内的细胞-材料复合物,见颜色变白,组织硬度增加,组织学和电镜观察见有大量骨组织形成。免疫组化示细胞为具有分泌特异性骨钙蛋白的成骨细胞。结论骨髓基质干细胞经成骨诱导为成骨细胞后与小肠粘膜下层共培养,植入裸鼠体内后可形成骨组织,小肠粘膜下层是一种良好的骨组织工程支架材料。  相似文献   

5.
目的研究兔骨髓基质细胞体外向成骨细胞分化的生物学特性及其与牛煅烧骨体外复合培养的生物相容性.方法将体外培养1周的兔骨髓基质细胞向成骨细胞诱导分化,于1、2、4周时提取RNA,采用RT-PCR技术检测ALP和I型胶原mRNA表达,并对培养2周的细胞行VonKossa染色观察钙结节形成;另制备细胞-煅烧骨复合物,继续培养1、7、14 d后取出,扫描电镜观察及X射线衍射仪检测.结果体外诱导培养2、4周后,兔骨髓基质细胞成功表达ALT和I型胶原,VonKossa染色可见钙结节形成.扫描电镜及X射线衍射分析证实细胞在煅烧骨表面大量贴附成活,14 d后细胞铺满支架表面,合成并分泌胶原纤维及钙盐.结论兔骨髓基质细胞体外可成功向成骨细胞诱导分化,成骨特性表达佳;与牛煅烧骨体外复合培养显示良好的生物相容性.  相似文献   

6.
[目的]评价异体冻干颗粒骨体外生物相容性,为其作为骨组织工程支架材料的临床应用提供实验依据。[方法]4周龄SD大鼠,雌雄不限,采用全骨髓贴壁法体外培养骨髓基质干细胞(BMSCs),取生长状态良好的第3代BMSCs行成骨诱导培养并鉴定。制备异体冻干颗粒骨浸提液,将成骨诱导7d的BMSCs加入浸提液作为实验组,未加浸提液的为对照组,培养1、3、5d,采用MTT法检测异体冻干颗粒骨浸提液对细胞增殖的影响。成骨诱导7d的BMSCs按1×106/ml接种于异体冻干颗粒骨上,倒置相差显微镜、扫描电镜观察与其复合培养的细胞在材料上的生长情况;酶消化法消化复合于冻干颗粒骨上的成骨细胞,流式细胞仪检测细胞周期变化。[结果]BMSCs成骨诱导7d,ALP染色示胞浆内可见阳性蓝染颗粒,胞核呈红色。MTT法检测结果显示细胞在浸提液中生长与增殖状态良好,实验组与对照组差异无统计学意义(P0.05);倒置相差显微镜可见颗粒骨表面粘附多数成骨细胞;扫描电镜观察可见细胞在材料表面粘附、伸展并能增殖、分泌产生细胞外基质;流式细胞术检测显示,异体颗粒骨对细胞周期无影响,实验组细胞均为正常2倍体细胞。实验组与对照组差异无统计学意义(P0.05)。[结论]异体冻干颗粒骨无细胞毒性并具有良好的组织细胞相容性,为其临床应用提供了实验依据。  相似文献   

7.
成人骨髓源成骨细胞体内异位成骨的实验研究   总被引:2,自引:1,他引:1  
目的 观察成人骨髓源成骨细胞与珊瑚羟基磷灰石 (CHA)复合构建的组织工程化骨组织的体内异位成骨能力 ,探讨适宜的组织工程化骨组织的构建方式。 方法 抽取健康成人骨髓 ,采用全骨髓法培养 ,使成人骨髓基质干细胞 (hBMSCs)定向诱导分化为成骨细胞 ,然后种植于CHA上 ,复合培养 5d后植入裸鼠股部肌袋内 ,以未种植细胞的CHA作为对照。术后 4,8,12周取材作一般观察、X线摄片、组织学检查和源于成人的碱性磷酸酶 (ALP)及骨钙素 (OCN)的RT PCR检测。 结果 原代和传代培养的细胞具有活跃的增殖能力 ,成骨细胞与CHA复合生长良好。实验组术后 4、8、12周均有新骨形成 ,随着时间延长 ,新骨生成量增多 ;对照组则均无新骨形成。术后 4周实验组RT PCR检测源于人的ALP及OCN表达均为阳性 ,对照组阴性。 结论 成人骨髓源成骨细胞与CHA复合培养后构建的组织工程化骨组织 ,具有良好的异位成骨能力 ,可望应用于临床修复骨缺损。  相似文献   

8.
[目的]观察成骨生长肽(osteogenic growth peptid,OGP)基因转染兔骨髓基质干细胞后的表达及表达产物对骨髓基质干细胞向成骨细胞分化的影响.[方法]构建重组真核表达载体pcDNA_(3.1)-OGP,并在脂质体介导下,将其导入兔骨髓基质干细胞.通过G418筛选获得阳性克隆.用RT-PCR方法榆测OGP基因在骨髓基质干细胞内的表达.Ⅰ型胶原和碱性磷酸酶的检测观察转染pcDNA_(3.1)-OGP后骨髓基质干细胞向成骨细胞分化情况.[结果]成功构建真核表达载体pcDNA_(3.1)-OGP,用RT-PCR方法及碱性磷酸酶和Ⅰ型胶原检测证实OGP基因能在骨髓基质干细胞中表达并促进其向成骨细胞分化.[结论]经pcDNA_(3.1)-OGP转染的兔骨髓基质干细胞不仅可以表达OGP,而且具有向成骨细胞系分化的特性.  相似文献   

9.
目的 观察骨髓来源的成骨细胞与新型双相陶瓷材料复合培养后细胞生长状况及其体内异位成骨能力。方法 将来源于兔骨髓的成骨细胞与新法制备的HA/TCP共培养2周,通过相差显微镜、扫描电镜观察体外细胞生长情况;然后将复合物自体异位植入体内,6周后观察体内成骨情况。结果 扫描电镜显示材料表面和孔隙内均有成骨细胞生长,有良好增殖活动性和稳定细胞表型,材料中心区未见细胞生长;植入体内6周后取材见内植物有新骨形成、多位于内植物表面,可见成骨细胞、骨细胞、髓腔样结构、板层样骨基质等正常骨组织结构。结论 新型双相陶瓷支架可用作组织工程骨的细胞外基质材料,骨髓来源的成骨细胞可用作骨组织工程的种子细胞。  相似文献   

10.
在体内组织中,有两类生成骨细胞的前体细胞:成骨前体细胞(DOPC)和可诱导的成骨前体细胞(IOPC)。前者主要存在于骨髓的基质细胞中,具有干细胞的特点,能自身复制分化成一定类型的细胞,此种基质细胞可分化成为成骨细胞、成纤维细胞、网状细胞等,即基质细胞中只有一部分属于骨祖细胞,能自发地分化为成骨细胞,骨髓多能干细胞在  相似文献   

11.
Background : We investigated the vasopressor hormone response following mesenteric traction (MT) with hypotension due to prostacyclin (PGI2) release in patients undergoing abdominal surgery with a combined general and epidural anesthesia. Methods : In a prospective, randomized, placebo-controlled study we administered 400 mg ibuprofen (i.v.) in 42 patients scheduled for abdominal surgery. General anesthesia was combined with epidural anesthesia (T4-L1). Before as well as 5, 15, 30, 45, and 90 min after MT we recorded plasma osmolality, hemodynamics and measured 6-keto-PGFlα (stabile metabolite of PGI2), TXB2 (stabile metabolite of thromboxane A2) active renin, and arginine vasopressin (AVP) plasma concentrations by radioimmunoassay. Catecholamine levels were assessed by high-pressure liquid chromatography (HPLC) with electrochemical detection. Results : Following MT, arterial hypotension occurred along with a substantial PGI2 release. This was completely abolished by ibuprofen administration. Although plasma levels of 6-keto-PGF (1133 (708) vs. 60 (3) ng/L, median (median absolute deviation), P=0.0001, placebo vs. ibuprofen) remained significantly elevated, blood pressure was restored within 30 min after MT in the placebo group. At the same point in time plasma concentrations of TXB2 (164 (87) vs. 58 (1) ng/L, P=0.0001), epinephrine (46 (33) vs. 14 (6) ng/L, P=0.001), AVP (41 ± (18) vs. 12 (7) ng/L, P=0.0004), and active renin (27 (12) vs. 12 (4) ng/L, P = 0.001) were significantly higher in placebo-treated patients. Conclusion : Under combined general and epidural anesthesia arterial hypotension following MT due to endogenous PGI2 release is associated with enhanced release of AVP, active renin, epinephrine and thromboxane A2, presumably contributing to hemodynamic stability within 30 min after MT.  相似文献   

12.
Don Dame 《Artificial organs》1996,20(5):613-617
Abstract: Virtually all blood pumps contain some kind of rubbing, sliding, closely moving machinery surfaces that are exposed to the blood being pumped. These valves, internal bearings, magnetic bearing position sensors, and shaft seals cause most of the problems with blood pumps. The original teaspoon pump design prevented the rubbing, sliding machinery surfaces from contacting the blood. However, the hydraulic efficiency was low because the blood was able to "slip around" the rotating impeller so that the blood itself never rotated fast enough to develop adequate pressure. An improved teaspoon blood pump has been designed and tested and has shown acceptable hydraulic performance and low hemolysis potential. The new pump uses a nonrotating "swinging" hose as the pump impeller. The fluid enters the pump through the center of the swinging hose; therefore, there can be no fluid slip between the revolving blood and the revolving impeller. The new pump uses an impeller that is comparable to a flexible garden hose. If the free end of the hose were swung around in a circle like half of a jump rope, the fluid inside the hose would rotate and develop pressure even though the hose impeller itself did not "rotate"; therefore, no rotating shaft seal or internal bearings are required.  相似文献   

13.
Abstract: A variety of protein-bound or hydrophobic substances, accumulating as a result of pathologic conditions such as exogenous or endogenous intoxications, are removed poorly by conventional detoxification methods because of low accessibility (hemodialysis), insufficient adsorption capabilities (hemosorption), low efficiency (peritoneal dialysis), or economic limitations (high-volume plasmapheresis). Combining advantages of existing methods with microspheric technology, a module-based system was designed. Major operating parameters of the latter can be modified to allow for adjustment to individual clinical situations. An extracorporeal blood circuit including a plasmafilter is combined with a secondary high-velocity plasma circuit driven by a centrifugal pump. Different microspheric adsorbers can be combined in one circuit or applied in sequence. Thus, a prolonged treatment can be tailored using specially designed selective adsorber materials. Comparing this system with existing methods (high-flux hemodialysis, molecular adsorbent recycling system), results from our in vitro studies and animal experiments demonstrate the superior efficiency of substance removal.  相似文献   

14.
Background : Our objective was to determine whether administration of propranolol or verapamil modifies the hemodynamic adaptation to continuous positive-pressure ventilation (CPPV), in particular the regional distribution of cardiac output (CO).
Methods : General hemodynamics and regional blood flows assessed by microsphere technique (15 (μm) were recorded in 16 anesthetized pigs during spontaneous breathing (SB) and CPPV with 8 cm H2O end-expiratory pressure (CPPV8) before and after intravenous administration of propranolol (0.3 mg · kg−1 followed by 0.15 mg · kg−1 · h−1, n=8) or verapamil (0.1 mg · kg−1 followed by 0.3 mg · kg−1 · h−1, n=8).
Results : CPPV8 depressed CO by 25% without shifts in its relative distribution with the exception of a noteworthy increase in adrenal perfusion. Propranolol increased arterial blood pressure, and due to a fall in heart rate, CO dropped by 25%. The kidneys and, to a lesser extent, the splanchic region and central nervous system received increased fractions of the remaining CO at the expense of skeletal muscle flow. Similar patterns were seen during SB and CPPV8 such that the combination of propranolol and CPPV8 depressed CO by 50%. The circulatory effects of verapamil were less evident but myocardial perfusion tended to increase.
Conclusions : The combination of propranolol or verapamil with CPPV does not result in any specific hemodynamic interaction in anesthetized pigs, except that the combined effect of propranolol and CPPV may severely reduce CO.  相似文献   

15.
Background: Obesity is increasing globallly, including in the formerly "Eastern Bloc" countries. Methods: A survey was made of obesity and bariatric surgery. Results: In the 8 East and Central European countries studied, with total population 300 million, roughly 43% of the population was overweight (BMI 25-30), 23% obese (BMI > 30), with about 15 million people morbidly obese (BMI > 40). From 0-10 morbidly obese individuals/100,000/year undergo bariatric surgery. Conclusion: Most countries were found to provide inadequate treatment for obesity.The majority of the morbidly obese are not treated effectively. However, health-care awareness of obesity and bariatric surgeons are slowly increasing.  相似文献   

16.
Background : Inhibitory effects of volatile anaesthetics on platelet aggregation have been demonstrated in several studies. However, the influence of volatile anaesthetics on intracoronary platelet adhesion has not been elucidated so far.
Methods : Isolated hearts of guinea pigs were perfused with buffer in the absence or presence of volatile anaesthetics (0.5 and 1 MAC) at constant coronary flow rates of 5 ml/min for 25 min, then 1 ml/min for 30 min and again 5 ml/min for 10 min. Before, during and after low-flow perfusion, a bolus of human platelets was applied into the coronary system. To simulate thrombogenic conditions, 0.3 U/ml human thrombin was infused during low-flow perfusion and reperfusion. The number of platelets sequestered to the endothelium was calculated from the difference between coronary in- and output of platelets. The myocardial production of lactate and consumption of pyruvate and coronary perfusion pressure were also determined.
Results : At a flow rate of 5 ml/min only about 3% of the applied platelets did not emerge from the coronary system, in any group. In contrast, 13.1±1.2% (mean±SEM) of infused platelets became adherent in low-flow perfusion in the control group without anaesthetic. The adherence was reduced with each 1 MAC isoflurane (to 6.2±1.2%), sevoflurane (to 4.4±0.9%) or halothane (to 3.2±1.5%) (each P <0.05 vs. control). Volatile anaesthetic, 0.5 MAC, did not inhibit platelet adhesion to a statistically significant extent in any case. Perfusion pressure and metabolic parameters were not statistically different between the control and the hearts exposed to anaesthetics.
Conclusion : Volatile anaesthetics in a concentration of 1 MAC can reduce the adhesion of platelets in the coronary system under reduced flow conditions. This action does not arise from vasodilation or inhibition of ischaemic stress.  相似文献   

17.
Background: It has been shown that the depressive effects of both propofol and midazolam on consciousness are synergistic with opioids, but the nature of their interactions on other physiological systems, e. g. respiration, has not been fully investigated. The present study examined the effect of propofol and midazolam alone and in combination with fentanyl on phrenic nerve activity (PNA) and whether such interactions are additive or synergistic. Methods: PNA was recorded in 27 anaesthetised and artificially ventilated rabbits. In three groups, propofol, fentanyl and midazolam were administered intravenously in incremental doses to construct dose-response curves for the depressant effects of each one on PNA. In another two groups, the effect of pretreatment with either fentanyl 1 μg · kg?1 i. v. or midazolam 0.05 mg · kg?1 i. v. on the effects of propofol and fentanyl respectively on PNA were studied. Results: Propofol and fentanyl caused a dose-dependent depression of PNA with complete abolition at the highest total doses of 16 mg · kg?1 i. v. and 32 μg · kg?1 i. v., respectively. In contrast, midazolam in incremental doses to a total of 0.8 mg · kg?1 reduced mean PNA by 63%, but approximately 12% of PNA remained at a total dose as high as 6.4 mg · kg?1. The mean ED50s, calculated from dose-response curves, were 5.4 mg · kg?1, 3.9 μg · kg?1 and 0.4 mg · kg?1 for propofol, fentanyl and midazolam, respectively. Initial doses of either fentanyl 1 μg · kg?1 i. v. or midazolam 0.05 mg · kg?1 i. v. acted synergistically with subsequent doses of either propofol or fentanyl to abolish PNA at total doses of 8 mg · kg?1 and 8 μg · kg?1, respectively. Conclusion: Fentanyl has a synergistic interaction with both propofol and midazolam on PNA and hence potentially on respiration.  相似文献   

18.
Background: Catecholaminergic support is often used to improve haemodynamics in patients undergoing major abdominal surgery. Dopexamine is a synthetic vasoactive catecholamine with beneficial microcirculatory properties. Methods: The influence of perioperative administration of dopexamine on cardiorespiratory data and important regulators of macro- and microcirculation were studied in 30 patients undergoing Whipple pancreaticduodenectomy. The patients received randomized and blinded either 2 μg · kg?1 · min?1 of dopexamine (n=15) or placebo (n=15, control group). The infusion was started after induction of anaesthesia and continued until the morning of the first postoperative day. Endothelin-1 (ET-1), vasopressin, atrial natriuretic peptide (ANP), and catecholamine plasma levels were measured from arterial blood samples. Measurements were carried out after induction of anaesthesia, 2 h after onset of surgery, at the end of surgery, 2 h after surgery, and on the morning of the first postoperative day. Results: Cardiac index (CI) increased significantly in the dopexamine group (from 2.61±0.41 to 4.57±0.78 1 · min?1 · m?2) and remained elevated until the morning of the first postoperative day. Oxygen delivery index (DO2I) and oxygen consumption index (VO2I) were also significantly increased in the dopexamine group (DO2I: from 416±91 to 717±110 ml/m2 · m2; VO2I: from 98±25 to 157±22 ml/m2 · m2), being significantly higher than in the control group. pHi remained stable only in the dopexamine patients, indicating adequate splanchnic perfusion. Vasopressive regulators of circulation increased significantly only in the untreated control patients (vasopressin: from 4.37±1.1 to 35.9±12.1 pg/ml; ET-1: from 2.88±0.91 to 6.91±1.20 pg/ml). Conclusion: Patients undergoing major abdominal surgery may profit from prophylactic perioperative administration of dopexamine hydrochloride in the form of improved haemodynamics and oxygenation as well as beneficial influence on important regulators of organ blood flow.  相似文献   

19.
Background: Halothane inhibits in vitro and in vivo activity of cytochrome P-450 (CYP) 2E1. There are several fluorinated volatile anaesthetics besides halothane, and most of them are defluorinated by CYP2E1. It is unclear whether other fluorinated anaesthetics inhibit the in vivo activity of CYP2E1.
Methods: We compared the inhibitory effects of therapeutic concentrations of four inhalational anaesthetics, halothane, enflurane, isoflurane, and sevoflurane, on chlorzoxazone metabolism in rabbits receiving artificial ventilation.
Results: All four inhalational anaesthetics decreased arterial blood pressure and increased plasma chlorzoxazone concentration. However, no significant differences in the plasma chlorzoxazone concentration were found between the four anaesthetics. The estimated chlorzoxazone clearance increased after beginning inhalation with all four agents, but no significant difference in clearance was noted between agents.
Conclusions: At therapeutic concentrations, the in vivo inhibitory effect on chlorzoxazone metabolism was similar for all four inhalational anaesthetics examined, even though their chemical characteristics and extent of hepatic metabolism differ considerably.  相似文献   

20.
A concept of balanced analgesia using nonsteroidal anti-inflammatory drugs (NSAIDs), paracetamol (acetaminophen), opioids, and corticosteroids can also be used in patients with pre-existing illnesses. NSAIDs are the most effective treatment for acute pain of moderate intensity in children; however, these drugs should be avoided in patients at increased risk for serious side effects, e.g. patients with renal impairment, bleeding tendency, or extreme prematurity. NSAIDs can be given with minimal risks to the younger child with mild to moderate asthma, and, in these patients, the use of steroids can be encouraged; in addition to their antiemetic and analgesic action, a beneficial effect on asthma symptoms can be expected. In the non-intubated child with cerebral trauma, exaggerated sedation caused by opioids and increased bleeding tendency caused by NSAIDs must be avoided. In neonates and small infants, the oral administration of sucrose or glucose is helpful to minimize pain reaction during short uncomfortable interventions.  相似文献   

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