Icariin suppresses bone resorption activity of rabbit osteoclasts in vitro

The effect of icariin on the bone resorption activity of rabbit osteoclasts is assessed in vitro. Osteoclasts were isolated from Japanese white rabbits and cultured on plates with a sterilized bone slice in each well. After treatment with icariin at various concentrations, the bone resorption activity of osteoclasts was evaluated by examining pit areas, superoxide anion (·O2-) generation, size and number of actin rings and intracellular calcium concentration [Ca2 ]i. As revealed by these data, icariin elicited continuous decline of [Ca2 ]i, making actin ring constricted and ·O2- generation decreased. These events resulted in smaller and fewer pits which indicate suppressed bone resorption activity of rabbit osteoclasts by icariin.     

Preparation of bioactive glasses with controllable degradation behavior and their bioactive characterization

Bioactive glasses and ceramics have been widely investigated for bone repair because of their excel-lent bioactive characteristics. However, these biomaterials undergo incomplete conversion into a bone-like material, which severely limits their biomedical application. In this paper, borosilicate bioac-tive glasses were prepared by traditional melting process. The results showed that borosilicate glasses possessed high biocompatibility and bioactivity. In addition, when immersed in a 0.02 mol/L K2HPO4 solution, particles of a borate glass were fully converted to HA. The desirable conversion rate to HA may be achieved through the adjustment of the B2O3/SiO2 ratio. The results of XRD and FTIR analysis indicated that the degradation product was carbonate-substituted hydroxyapatite, which was similar to the inorganic component of bone.     

细菌纤维素组织工程支架的仿生矿化研究

骨骼创伤已经成为当今影响人类健康的一大病症。因此,骨修复材料就成为研究的一大热点。骨组织工程支架作为重要的骨修复材料,可以诱导成骨细胞生长并为新骨生长提供条件。传统的骨组织工程支架包括合成高分子(如聚乳酸、聚乙醇酸等)和天然高分子(如胶原、壳聚糖等)。与传统支架材料相比,细菌纤维素(BC)具有良好的生物相容性、精细的纳米空间三维网络结构,有作为组织工程支架的潜能。通过仿生矿化处理,BC纳米纤维表面可以生长出羟基磷灰石(HA)的晶体颗粒,且HA颗粒均匀覆盖在纳米纤维表面。通过热分析得出,仿生矿化处理会使BC的热稳定性得到一定的提升。     

羟基磷灰石基生物材料培养破骨细胞研究现状

骨重建是新骨组织替换旧骨或受损骨的生理过程，在无瘢痕骨愈合和受损骨再生中起着必要的作用。骨重建主要是骨形成细胞如成骨细胞（osteoblast，OB），骨吸收细胞如破骨细胞（osteoclast，OC），和巨噬细胞（macrophage，MP）等细胞之间的相互协调活动。OC作为生物体内唯一多核细胞，负责生物体内骨质的脱钙与骨基质的吸收。自然骨具有一定硬度、韧性，良好的生物活性，保证骨重建中细胞的增殖、分化、骨形成或骨再吸收等活性。以羟基磷灰石（hydroxyapatite，HA）为主的磷酸钙（HA-CaPs）是自然骨中矿物质的主要成分，具有优异的生物性能而被广泛应用于骨组织修复领域。总结了HA-CaPs物相成分、表面形貌等对OC等骨吸收细胞增殖、分化和骨吸收活性的影响，并通过OC对于骨修复材料体内吸收机制及细胞活性调控机制等探讨HA-CaPs与细胞间相互关系，以期为磷酸钙人工骨替代材料更加广泛的生物应用作理论参考。     

Synergistic effect of trace elements and flavonoids from Epimedium koreanum Nakai on primary osteoblasts

Several trace elements, particularly, manganese （Mn） and zinc （Zn）, are essential in bone metabolism as cofactors for specific enzymes. It has been reported that there exists the relationship between osteoporosis and trace element-deficiency and the efficacy of Ca, Mn and Zn supplementation on spinal bone mineral density in postmenopausal women. Traditional Chinese medicines （TCM）, such as Herba epimedii, were proved to be effective for prevention of osteoprosis in vivo; however, the efficacy of the main constituents and/or crude extract was not ideal in vitro, which suggested that they may work in another way. The purpose of the present study was to examine whether the combination of icariin and total flavonoids （TF） from Herba epimedii with mineral elements, which were abundant in Herba epimedii, would have a more beneficial effect on the viability and differentiation of primary osteoblasts than either agent alone, and to analyze the dada for a possible synergistic, additive or antagonistic effect. The combinations of 10 μmol/L Zn, Ca and Mn with icariin and total flavonoids greatly improved the cell viability and meanwhile dramatically enhanced the alkaline phosphatase activity as compared to each agent alone. On the other hand, an increased cell growth inhibition was also observed by combining 0.1 μmol/L, 1 pmol/L Zn with 10μmol/L icariin, and 10 μmol/L Mn with 0.06 μg/mL total flavonoids. Meanwhile a decreased alkaline phosphatase activity was also found in several icariin-Zn/Mn and total flavonoids-Zn/Ca/Mn combinations. These results suggested that mineral elements （Zn, Ca, Mn） greatly enhanced the efficacy of icariin and total flavonoids from Herba epimedii on the viability and differentiation of primary osteoblasts by certain combinations.     

人工骨修复材料磷酸八钙纤维的合成

磷酸八钙（OCP）纤维具有优良的生物活性和力学性能,可广泛应用于骨替换和骨修复。本文采用均相沉淀法制备了磷酸八钙纤维,并通过红外光谱、XRD衍射、扫描电镜对产物进行表征、探讨了反应体系初始pH值、反应温度及体系pH值调节剂尿素浓度等因素对合成磷酸盐的组成及其形貌的影响。实验发现合成条件对磷酸八钙形貌及产物组成的影响较大：尿素浓度过低,所得产物形貌的长短和粗细分布极不均匀;尿素浓度偏高时,产物较为粗短,且产物为OCP和羟基磷灰石（HA）的混合磷酸盐;体系反应温度的提高有利于OCP的快速析出和产率的提高,所得产物也更为纤细。随着体系初始pH值的升高,所得磷酸盐的形貌逐渐变得粗短,且不均匀,不利于生成OCP,产物有向HA转化的趋势。研究表明,当Ca(NO3)2为0.167mol/L,(NH4)2HPO4为0.1mol/L,(NH2)2CO为0.6～0.7mol/L,体系初始pH值在2.2左右,反应温度为90℃时,可制得纤细的磷酸八钙纤维。     

碳酸钠液相改性硅-磷酸钙复合骨水泥研究

以质量分数70%的硅酸三钙(Ca3SiO5,C3S)和30%磷酸氢钙(CaHPO4·2H2O,DCPD)复合得到的DCP30粉体材料为固相,以不同浓度碳酸钠溶液为液相,得到碳酸钠改性骨水泥材料。使用X射线衍射(XRD)、扫描电镜(SEM)、万能材料试验机等手段对不同浓度改性材料进行表征。结果显示:添加碳酸钠液相,骨水泥初、终凝时间分别缩短至16和55min;调控碳酸钠液相浓度,可以实现短期抗压强度优化;使用碳酸钠后,固化自发生成羟基磷灰石(HA)。浸泡模拟体液(SBF)7天,材料表面覆盖HA沉积层,生物活性优越。碳酸钠液相改性硅-磷酸钙复合骨水泥体系的水化性能、短期力学性能以及生物活性均优于Ca3SiO5水泥和未改性硅-磷酸钙复合骨水泥,是一种良好的生物活性骨修复材料。     

水热法合成含锌羟基磷灰石

自然骨中的无机组分并不是纯的羟基磷灰石[（Ca10（PO4）6（OH）2,Hydroxyapatite,简称HA],还含有微量镁离子（Mg2＋）、锌离子（Zn2＋）等。为满足临床的需要,常常需要在HA中添加锌元素制成含锌羟基磷灰石（Zn-HA）,其有望成为新一代的骨修复和替换材料。以硝酸钙、磷酸三铵、硝酸锌为原料,在210℃下水热处理8h合成Zn-HA和Ca19Zn2（PO4）14混合粉末。采用X射线衍射（XRD）、电子扫描电镜（SEM）和热重—差热分析（TG-DTA）法对合成粉体的晶相、形貌和热稳定性进行分析。结果表明：在210℃、8h的实验条件下,反应釜的填充度较大时容易合成Zn-HA,但Zn掺量的摩尔分数不宜超过4%;随着Zn掺量和反应釜填充度的增加,HA的结晶度下降;Zn的掺入促使HA易分解,使其热稳定性较差。     

Comparison of Compositions and Structural Characterizations of Synthetic Nano-hydroxyapatite and Mineral Phase from Porcine Femoral Bone

In the paper, nano-hydroxyapatite was synthesized by hydrothermal treatment. The erystal morphology and compositions of the synthetic HA were investigated using TEM,IR, XRD and XPS techniques and were compared with those of porcine femoral bone apatite. The results showed that the synthetic HA is nanometer rod crystal similar to the bone apatite, and the average crystal sizes calculated by Scherrer equation for the four samples (HA dried at 80℃, unburned bone apatite, HA calcined at 750℃ and bone apatite calcined at 750℃ )are 20.6, 21.7, 22.5 and 27.6nm ,respectively, Moreover, the essential compositions for the synthetic HA and bone apatite are almost the same.     

淫羊藿甙及其与铁离子配合物的电喷雾质谱研究

本文利用多级串联电喷雾质谱 (ESI-MSn)研究了淫羊藿甙及其与铁离子形成的配合物 ,得到了淫羊藿甙的质谱碎裂规律 ;并对淫羊藿甙与铁离子形成的配合物结构进行了初步确定。结果表明 :淫羊藿甙与铁离子可形成比较稳定的 2 :1的配合物     

淫羊藿苷与过氧化氢酶相互作用的荧光光谱法和分子对接研究

利用过氧化氢酶的内源荧光,通过荧光光谱法对淫羊藿苷与过氧化氢酶的相互作用进行了研究.结果表明,淫羊藿苷主要以静电作用力与过氧化氢酶发生相互作用,16℃时结合常数和结合位点数分别为4.59×104 L/mol和1.00.根据F（o）rster非辐射能量转移理论,计算二者相互作用的结合距离为4.05 nm.对二者之间的相互作用进行了分子模拟,表明淫羊藿苷在过氧化氢酶4个亚基的中心区域通过静电作用力、疏水作用力和氢键作用力与过氧化氢酶发生相互作用,与荧光光谱法的研究结果相一致.     

利塞膦酸盐对磷硅酸钙理化及生物性能的影响

将利塞膦酸盐(0.5%和1.0%)与含有磷硅酸钙的骨水泥粉末混合后进行水化反应,测定骨水泥固化时间及抗压强度,并分析其载药前后的微观结构变化.通过细胞毒性与基因表达水平检测分析了骨水泥的生物性能.结果表明:在0.5%药物水平下,骨水泥促进成骨细胞增殖与分化效果最佳.药物可延缓骨水泥固化,但骨水泥的气孔率会随之降低且晶体结构会更加紧密.载有利塞膦酸的磷硅酸钙骨水泥具有与松质骨相似的力学强度及良好的生物相容性,作为骨组织修复材料具有很好的应用价值.     

Maturation of Osteoblast-Like Saos-2 on Hydroxyapatite Ceramics

Hydroxyapatite ceramics (.HA) has been proved to be excellent in biocompatibility and bioactivity.However, limited information is available concerning how HA ceramics affects the maturation of es-tcoblasts in molecular biological level /n v/tro. This study examines the mRNA expression and protein production of hone-related genes in estcoblast-like cell line(Saos-2) cultured on HA disks. Saos-2 cells are seeded onto the substrates and cultured for 18 days. Harvested cells are tested for the cell growth rate, expression of mRNAs for esteocalcin and alkaline phosphatase, and protein production of bone sialo-protein and esteocalcin. MTS assay shows that cell proliferates well on HA ceramic substrate. After 9d,bone sialoprotein and esteocalcin protein production in SAPS-2 increases more on HA surfaces than on control material. As bone sialoprotein and esteocalcin are the genes to be highly expressed at the late stage of estcoblast differentiation, this study reveals that after long time culture in HA, HA can induce Saos-2 maturation. The behavior of Saos-2 on HA surfaces revealed in this study provides valuable infor-mation for the understanding of the biocompatibility and bioactivity of HA ceramics.     

纳米羟基磷灰石和中药结合在骨修复材料中的应用

探讨了几种具有能促进骨质再生和愈合、加速成骨细胞活化和骨基质钙化等疗效的中药,随着生物材料的发展,有望在骨修复材料中加入促进骨愈合的中药制剂来促进骨的生长,同时这些中药还有消炎、抗菌的功效,达到了局部给药的目的,减少炎症的发生.     

Comparative study of microstructural remodification to porous β-TCP and HA in rabbits

To assess the remolding ability of repaired bone in hydroxyapatite （HA） and β-calcium phosphate （β-TCP） scaffold, two 75% porosity bioceramics with the same three-dimensional geometry were implanted into femoral condyles of rabbits. Histological and micro-computed tomography （micro-CT） results demonstrated abundant new bone formation in the porous HA scaffold along with indistinctive scaffold degradation. Results also indicated that scaffold resorption in the β-TCP group, which was followed by a replacement with newly formed bone, was significantly higher than that in the HA group. The crosslinking trabeculae remodeled from the mixtures of the newly formed bone and β-TCP scaffold remnants might be helpful to promoting even loading and reducing stress. The bone remodeling pattern resulted from bone formation and scaffold resorption was significantly different for the two bioceramics. The results demonstrated that the 75% porous β-TCP was more suitable for new bone remodification than HA scaffold.     

Hydroxyapatites enriched in silicon – Bioceramic materials for biomedical and pharmaceutical applications

Hydroxyapatite (Ca10(PO4)6(OH)2, abbreviated as HA) plays a crucial role in implantology, dentistry and bone surgery. Due to its considerable similarity to the inorganic fraction of the mineralized tissues (bones, enamel and dentin), it is used as component in many bone substitutes, coatings of metallic implants and dental materials. Biomaterial engineering often takes advantage of HA capacity for partial ion substitution because the incorporation of different ions in the HA structure leads to materials with improved biological or physicochemical properties. The objective of the work is to provide an overview of current knowledge about apatite materials substituted with silicon ions. Although the exact mechanism of action of silicon in the bone formation process has not been fully elucidated, research has shown beneficial effects of this element on bone matrix mineralization as well as on collagen type I synthesis and stabilization. The paper gives an account of the functions of silicon in bone tissue and outlines the present state of research on synthetic HA containing silicate ions (Si-HA). Finally, methods of HA production as well as potential and actual applications of HA materials modified with silicon ions are discussed.     

自固化载硒纳米微球/磷酸钙复合骨修复材料的制备及性能

研究制备具有促修复、抗肿瘤的可注射缓释硒纳米微球复合磷酸钙骨水泥材料.采用乳化交联法制备Na_2SeO_3/CS缓释硒纳米微球,将微球与磷酸钙骨水泥(CPC)复合,制备Na_2SeO_3/CS/CPC骨修复系统,对该系统的固化时间、力学强度、缓释硒及降解性能、形貌、晶相构成进行测定和表征分析,并对其体外细胞活性进行研究.结果表明:相对于纯的CPC,当掺入Na_2SeO_3/CS纳米微球的量为4%时,复合CPC的注射性良好,固化时间5.75～11.5 min,固化强度提高,微观结构显示CPC均匀地包裹在壳聚糖微球表面并形成了针片状HA晶体,微球的添加对复合CPC材料的晶相组分无显著影响,缓释硒效应良好,有效缓释达22 d,降解性优于纯CPC,形成了蜂窝状完善的网状三维立体多孔结构,利于组织细胞和血管及神经的黏附长入.体外细胞实验表明,复合CPC对人乳腺癌细胞MCF-7及MG-63人骨肉瘤细胞增生的抑制作用显著,且对两株细胞增生的抑制性差异不显著.本研究为非承骨微创、缺陷修复及骨肿瘤的术后恢复、防止复发和转移的预防和治疗提供一种新思路,同时为拓展功能元素硒的合理应用奠定实验基础.     

纳米羟基磷灰石对两种药物的载药性能研究

骨缺损常常伴随着炎症、肿瘤等情况,以各种骨修复材料为载体的药物缓释体系是一种新型的给药方式.以纳米羟基磷灰石(n-HA)为载药体系,用红外光谱和透射电镜表征了n-HA的结构,考察了其对抗生素阿莫西林和抗肿瘤药鬼臼毒素的不同吸附作用,并研究其对抗生素阿莫西林吸附量的影响.实验结果表明,n-HA的红外图谱及粒径与人体骨颇为接近,同时n-HA可以吸附有羟基的阿莫西林,并且吸附性能随着阿莫西林初始浓度的不同而不同,吸附速度快,4 h达到吸附平衡;但是对没有特征基团的鬼臼毒素,n-HA不能吸附.n-HA这种对有某些特征基团分子的特异性吸附作用,对载药骨材料的研究具有非常重要的指导意义.     

淫羊藿苷分子印迹聚合物吸附性能研究

以沉淀聚合法制备了淫羊藿苷分子印迹聚合物(MIPs).并用动态吸附、静态吸附和选择性吸附实验研究了制备的分子印迹聚合物对模板分子淫羊藿苷的吸附性能.结果表明制备的淫羊藿苷分子印迹聚合物对模板分子淫羊藿苷具有较好的吸附能力和识别能力,可以作为固相萃取材料来分离提取中草药淫羊藿中的淫羊藿苷.     

淫羊藿苷与脱水淫羊藿素的体外抗氧化研究

为研究淫羊藿苷和脱水淫羊藿素的体外抗氧化活性,分别测定了两者对1,1-二苯基-2-苦苯肼自由基(DPPH.)、超氧阴离子自由基(O2.)、羟自由基(OH.)的清除能力、对脂质过氧化的抑制能力、还原力以及总抗氧化能力.实验结果表明,淫羊藿苷具有很强的清除DPPH.、超氧阴离子自由基,还原力和总抗氧化的能力,有较强的清除OH.的能力,且均呈剂量依赖性.脱水淫羊藿素对DPPH.的清除能力很强,对OH.、超氧阴离子和脂质过氧化有较强的清除和抑制作用,表明淫羊藿苷和脱水淫羊藿素具有明显的抗氧化活性.