骨髓间充质细胞与速固化磷酸钙支架共培养的实验研究

[目的]观察人骨髓间充质干细胞在速固化磷酸钙骨水泥支架中的增殖和生长情况。[方法]将速固化壳聚糖-磷酸钙骨水泥支架材料预先成形。采用全骨髓法分离人骨髓间充质细胞并在体外对细胞进行培养和扩增。72h首次换液,细胞融合80%传代,取第3代细胞用于实验。将细胞与磷酸钙骨水泥支架复合培养,分别利用四甲基偶氮唑蓝MTT法及扫描电镜等检测细胞在材料中的增殖和生长情况。[结果]用酶标仪检测OD值提示共培养后,支架内的细胞数量逐渐增加。扫描电镜结果显示支架材料内部为多孔结构,孔内见有细胞生长,细胞表面可见分泌颗粒。[结论]速固化壳聚糖-磷酸钙骨水泥对人骨髓间充质干细胞有较好的细胞相容性,细胞可在材料内黏附和增殖,是理想的骨组织工程支架材料。     

大鼠骨髓间充质干细胞与多孔双相磷酸钙陶瓷支架体外黏附的实验研究

目的研究大鼠骨髓间充质干细胞(marrowmesenchymalstemcells,MSCs)诱导培养后在多孔双相磷酸钙(biphasiccalciumphosphate,BCP)陶瓷支架材料上的黏附和生长。方法SD大鼠MSCs经矿化诱导培养、扩增,具有成骨细胞表型后,与多孔BCP陶瓷支架及普通多孔羟基磷灰石(hydroxyapatite,HA)陶瓷支架体外复合培养,扫描电镜观察比较MSCs在两种材料支架表面的黏附数量和形态;同时以0.5、1.0、2.0、3.0和4.0×106/ml浓度细胞悬液接种于多孔BCP支架材料,检测适宜的接种浓度及单位体积支架材料可黏附MSCs数量。结果大鼠MSCs经诱导培养14d后,行矿化沉积茜素红染色、型胶原免疫细胞化学染色及碱性磷酸酶细胞化学染色,结果均为阳性。大鼠MSCs黏附于多孔BCP陶瓷上的细胞数(88.00±6.58)明显高于HA陶瓷组(39.00±3.65),两组比较差异有统计学意义(P<0.01)。当接种浓度为2.0×106/ml时,单位体积的陶瓷支架材料最多可黏附MSCs数量为1.28×107个/cm3,为细胞适宜接种浓度。结论大鼠MSCs在体外经矿化诱导培养可表达成骨细胞表型,细胞浓度为2.0×106/ml时与多孔BCP陶瓷支架材料具有良好的黏附能力。     

Micro-CT observation of rat dental pulp healing after pulpotomy in in vivo study

We report the newly developed Micro-CT, which allows us to observe the individual animal over a long experimental period and to compare changes in pulp tissue in relation to growth and aging without considering individual differences. Further, we used pathological examination to prove similar the result observing from Micro-CT. We have examined wound healing of teeth after pulpotomy in rats, and could clearly observe histopathological changes in the affected teeth and the absorption of temporary filling material and pulp capping agents. In cases with breakage of the dental crown, the CT images agreed with the pathological observations, and it was possible to estimate the time of breakage. In vivo Micro-CT is possible to apply in continuous recording of small experimental animals, such as rats, under anesthesia and the result is sufficiently high. High-quality image was obtained in of the entire head region of the rat. It was suggested that this method can be used for long-term continuous observation of changes in the teeth conditions after pulpotomy in experimental animals. We report the newly developed Micro-CT, which allows us to observe the individual animal over a long experimental period and to compare changes in pulp tissue in relation to growth and aging without considering individual differences.     

细胞内游离Ca2+在发育中大鼠皮层神经元无镁诱导惊厥性损伤中的作用

目的: 观察细胞内游离Ca2+([Ca2+]i)在培养的不同发育阶段皮层神经元无镁诱导惊厥性损伤中的作用,探讨惊厥性脑损伤年龄依赖性的可能机制.方法:体外培养6 d、17 d的胚胎大鼠皮层神经元用无镁细胞外液处理3 h,或于无镁处理前用NMDA(N-甲基-D-门冬氨酸)受体拮抗剂或Ca2+通道阻滞剂预处理,用MTT代谢率测定的方法检测神经元损伤,以Fluo-3作标记用激光共聚焦显微镜扫描的方法检测[Ca2+]i.结果:体外培养6 d、17 d的神经元单纯无镁组MTT代谢率较同期对照组降低.应用MK-801 10 μmol*L-1、AP-5 50 μmol*L-1、尼莫地平10 μmol*L-1预处理后再给无镁处理,培养6 d、17 d的神经元MTT代谢率均不同程度高于同期单纯无镁组.培养6 d、17 d的神经元相对荧光强度之间差异有显著性,两者与基线荧光强度比较差异亦有显著性.应用上述各种拮抗剂后,[Ca2+]i改变的峰值均明显低于同期单纯无镁组.结论: 在体外不同发育阶段的神经元,短暂无镁处理诱导惊厥样放电所引起的神经元线粒体功能损伤以及[Ca2+]i改变程度不同.这种[Ca2+]i改变的年龄依赖性可能是惊厥导致神经元损伤的年龄依赖性的机制之一.NMDA受体-Ca2+通道激活是导致这种[Ca2+]i改变及神经元损伤的关键环节.     

水泥型羟基磷灰石人工骨的制备及性能分析

Objective: To prepare hydroxyapatite cement (or calcium phosphate cement,CPC) and analyze its capability. Methods: Tetracalcium phospluge (TTCP ) was prepared by the method of high heat. TTCP reacted with in simulated body situation and produced CPC. Its capability was analyzed by scanning electron microscopy ( SEM), X-ray diffraction( XRD). Its density, absorbing water coefficient, macroporosity and compressive strength were measured also. Results: The main element of CPC is hydroxyapatile (HA), its microstructure comprised of petal crys-tals. The diameter of micropore was 4-10μm, density was 1. 922 g/cm^3, macroporosity was 29. 777%, absorbing coefficient was 15. 503%, compressive strength was 42.70 Mpa. Conclusion: This CPC has three-dimensional spatial structure, its strength meets the need of cancellous bone grafting.     

PLACENTAL CALCIUM PUMP: CLINICAL-BASED EVIDENCE

Placenta can be considered as a pump of calcium necessary for the normal development of the fetus. We believe that the location of this pump is in the placental basement membrane. The calcification of this membrane has been described only in cases of in utero fetal death. In this study we describe for the first time a case of placental calcification in a living fetus. The fetus of a normal 21-year-old pregnant woman showed heart abnormalities but the genetic analysis showed a normal male karyotype. The histology of the placenta demonstrated multiple intravillous linear and granular calcific incrustations The hemtoxylin/eosin stain of the sections revealed basement membrane calcific incrustations and intravillous calcium deposits. We postulate that the fetal circulation in the villi was impaired and the calcium that reached the villi from the mother was deposited at this level.     

Fumonisins as a possible contributory risk factor for primary liver cancer: A 3-year study of corn harvested in Haimen, China, by HPLC and ELISA

Employing HPLC fluorometry, gas-liquid chromatography (GLC) and a novel enzymelinked immunosorbent assay (ELISA) based on a monoclonal antibody, 40 corn samples, each collected in 1993 from agricultural stocks for human consumption in Haimen (Jiangsu County) and Penlai (Shandong Province), high- and low-risk areas for primary liver cancer (PLC) in China, respectively, were analysed for fumonisins (FBs), aflatoxins (AFs) and trichothecenes. Levels and positive rates of FBs and deoxynivalenol (DON) were significantly higher in Haimen than in Penlai. ELISA of the 40 corn samples harvested in the two areas in 1994 revealed that FB contamination levels and rates in these areas were comparable to those observed in 1993 in Haimen. ELISA analysis of 1993 and 1994 products revealed a wide occurrence of AFB₁ but the positive rates as well as levels were not significantly different between these areas. ELISA of the same sample number of corn harvested in 1995 revealed that FB contamination in Haimen was significantly higher than in Penlai. These 3-yearly surveys of corn samples (240 in total) demonstrated that corn harvested in Haimen was highly contaminated with FBs and that the contamination level, as well as positive rate in 1993 and 1995, were 10–50-fold higher than those in Penlai, suggesting FBs as a risk factor for promotion of PLC in endemic areas, along with the trichothecene DON. Co-contamination with AFs, potent hepatocarcinogens, was assumed to play an important role in the initiation of hepatocarcinogenesis.     

水泥型羟基磷灰石人工骨的制备及性能分析(英文)

目的:制备水泥型羟基磷灰石人工骨,并进行性能分析。方法:高温法制备出磷酸四钙,然后在模拟体内环境下将其与无水磷酸氢钙发生水化固化反应,合成水泥型羟基磷灰石人工骨,采用X线衍射、扫描电镜观察、体积密度、吸水率、孔隙率及耐压强度测量进行性能分析。结果:该人工骨主要成分为低结晶度的羟基磷灰石;微观结构呈针状或花瓣状晶体构成,微孔直径约4-10 μm,孔隙相互交通;平均体积密度为1.922 g·cm~(-3);平均吸水率为15.503％;孔隙率为29.777％;耐压强度为42.70 MPa。结论:本研究所制人工骨具有三维空间结构,力学性能符合人体非负重骨植骨要求。     

Artificial bone of porous tricalcium phosphate ceramics and its preliminary clinical application

Summary The authors have prepared the artificial bone of porous tricalcium phosphate ceramics according to an appropriate formula and manufacturing technology. Physical and chemical testing shows that it possesses several distinguishing features: the communicating pores and macro/micropores; mean pore size, 380 μm (from 240 μm to 510 μm); porosity, 46.4 %; and compressive strength, 97.4 kg/cm². It consists of CaO (49.09 %) and P₂O₅ (48.84 %). The testing of its biocompatibility shows that it is devoid of systemic or local toxicity, and free of irritation or foreign body response in tissues, and it does not result in hemolysis or mutation. The new bone readily grows into its pores with direct contact to the implanted material. 11 cases of bone defects were treated with this artificial bone with satisfactory results.     

预防性应用钙拮抗剂对缺血大鼠心肌细胞和红细胞内钙变化的影响

利用大鼠缺血模型观察预防性应用钙桔抗剂尼莫地平对大鼠缺血心肌和红细胞内钙的影响发现，缺血时大鼠心肌细胞、心肌组织和红细胞内均有钙积聚，红细胞膜钙泵活性下降；缺血前30分钟给予尼莫地平可使红细胞内钙恢复至正常水平，红细胞膜钙泵活性升高；心肌细胞和心肌组织内钙虽较缺血鼠降低，但仍高于对照组。提示：（1）缺血时细胞的钙内流可能是全身性的；（2）预防性应用尼莫地平可阻滞或减轻细胞内钙超载，同时还可保护细胞膜膜泵活性。