羟基磷灰石陶瓷诱导成骨细胞的研究

通过定量检测在羟基磷灰石(HA)表面生长的成骨细胞骨涎蛋白和骨钙蛋白的表达,研究了羟基磷灰石陶瓷对成骨细胞的影响．研究结果表明,随着成骨细胞培养时间的延长,在羟基磷灰石表面生长的成骨细胞骨钙蛋白和骨涎蛋白的表达量相对于对照组有显著增加．由于骨钙蛋白和骨涎蛋白是成骨细胞分化晚期高度表达的两种标志性蛋白质,因此其表达量的显著提高意味着羟基磷灰石陶瓷可以诱导成骨细胞的成熟。这一结果可进一步为理解HA陶瓷的生物相容性和生物活性提供分子生物学依据。     

钙硅摩尔比对硅钙石及磷结晶产品的影响

以电石渣和白炭黑为原料合成硅钙石,并以硅钙石为晶种结晶,考察了不同钙硅摩尔比对硅钙石及磷结晶产品结构的影响;研究发现钙硅比的差异对硅钙石及磷结晶产品的影响较大,钙硅比为1.75∶1时合成羟基硅钙石与磷酸盐反应后的磷结晶产品主要为晶型异常的羟基磷灰石晶体,而其他几种钙硅比下的磷结晶产品主要为不同形式的磷酸钙,不同的钙硅比导致了硅钙石结晶程度的差异,影响了其反应活性,进而影响硅钙石释放Ca2+和OH-的能力,因此得到不同结构的磷酸钙及羟基磷灰石;钙硅比为1.75∶1的条件下合成的羟基硅钙石最有利于以结晶为羟基磷灰石的形式从废水中回收磷。     

牛血清白蛋白-羟基磷灰石难溶性复合物的FT-IR光谱研究

利用富立叶变换红外光谱(FT-IR)法对牛血清白蛋白(BSA)-羟基磷灰石［Ca₁₀(OH)₂(PO₄)₆］难溶性复合物的组成及BSA与羟基磷灰石的作用方式进行了研究。结果表明,复合物中BSA与羟基磷灰石之间存在着强烈的相互作用,这种相互作用使水溶性的BSA进入固体羟基磷灰石的结构形成难溶性BSA-羟基磷灰石复合物。复合物具有非化学计量的性质,体现了生物矿化的特征。正是这种蛋白质与羟基磷灰石间的复杂相互作用,形成高级自组装结构,使矿化产物具有高强度和韧性。     

以碳酸钙为模板微波水热合成碳酸羟基磷灰石微球

通过将碳酸钙微球浸入到磷酸氢二铵溶液中,并在60℃条件下进行微波水热处理,获得了直径约为1μm的羟基磷灰石微球,其转化机理为溶解-再沉淀反应。碳酸钙微球浸入到磷酸氢二铵溶液中后,在微波辅助作用下,钙离子从碳酸钙表面释放并与溶液中的磷酸根离子进行反应,获得了羟基磷灰石微球,该微球保持了起始球霰石的形状和尺寸。本研究为可控构象的磷灰石基生物材料的制备提供了简便而有效的方法。     

马德隆常数的数值确定

离子晶体为数目巨大的多离子系统,其马德隆常数一般由实验确定。考虑晶体表面离子与内部离子对结合能贡献的不同,作者对马德隆常数定义公式进行修正,用程序模拟的办法确定了NaCl晶体的实验参数马德隆常数,并与实验确定值进行了比较。     

Crystal structure of the calcium pump of sarcoplasmic reticulum at 2.6 A resolution

Calcium ATPase is a member of the P-type ATPases that transport ions across the membrane against a concentration gradient. Here we have solved the crystal structure of the calcium ATPase of skeletal muscle sarcoplasmic reticulum (SERCA1a) at 2.6 A resolution with two calcium ions bound in the transmembrane domain, which comprises ten alpha-helices. The two calcium ions are located side by side and are surrounded by four transmembrane helices, two of which are unwound for efficient coordination geometry. The cytoplasmic region consists of three well separated domains, with the phosphorylation site in the central catalytic domain and the adenosine-binding site on another domain. The phosphorylation domain has the same fold as haloacid dehalogenase. Comparison with a low-resolution electron density map of the enzyme in the absence of calcium and with biochemical data suggests that large domain movements take place during active transport.     

Stem cell engraftment at the endosteal niche is specified by the calcium-sensing receptor

During mammalian ontogeny, haematopoietic stem cells (HSCs) translocate from the fetal liver to the bone marrow, where haematopoiesis occurs throughout adulthood. Unique features of bone that contribute to a microenvironmental niche for stem cells might include the known high concentration of calcium ions at the HSC-enriched endosteal surface. Cells respond to extracellular ionic calcium concentrations through the seven-transmembrane-spanning calcium-sensing receptor (CaR), which we identified as being expressed on HSCs. Here we show that, through the CaR, the simple ionic mineral content of the niche may dictate the preferential localization of adult mammalian haematopoiesis in bone. Antenatal mice deficient in CaR had primitive haematopoietic cells in the circulation and spleen, whereas few were found in bone marrow. CaR-/- HSCs from fetal liver were normal in number, in proliferative and differentiative function, and in migration and homing to the bone marrow. Yet they were highly defective in localizing anatomically to the endosteal niche, behaviour that correlated with defective adhesion to the extracellular matrix protein, collagen I. CaR has a function in retaining HSCs in close physical proximity to the endosteal surface and the regulatory niche components associated with it.     

Conformational changes associated with ion permeation in L-type calcium channels

The mechanism by which ions deliver their message to effector proteins involves a change in the protein conformation which is induced by the specific interaction of the ion with its binding site on the protein. In the case of an ion-channel protein, conformational changes induced by permeant ions and the consequences for channel function have received little attention. Here we report that binding of permeant cations to an intra-channel binding site of the dihydropyridine (DHP)-sensitive (L-type) Ca2+ channel leads to a conformational change which destabilizes the protonated state of a group on the external channel surface, and can shift its apparent pK value by more than 2 pH units. The lifetime of the protonated state correlates with the occupancy of an intra-channel binding site by the permeant cation. The demonstration of such conformational changes in a channel protein induced by the permeant ion has important implications for realistic models of the mechanism of ion permeation.     

Phytochemistry: structure of the blue cornflower pigment

The same anthocyanin pigment makes roses red but cornflowers blue, a phenomenon that has so far not been entirely explained. Here we describe the X-ray crystal structure of the cornflower pigment, which reveals that its blue colour arises from a complex of six molecules each of anthocyanin and flavone, with one ferric iron, one magnesium and two calcium ions. We believe that this tetrametal complex may represent a previously undiscovered type of supermolecular pigment.     

氨甲环酸与Wnt信号通路中含kringle结构域蛋白的结合对新生小鼠骨密度的影响

最近研究发现Wnt信号通路在骨形成过程中发挥重要作用Wnt受体如脂蛋白相关蛋白5（lrp5）和孤独受体（Ror2）的缺失或突变导致骨的不正常发育．Dkk是一个分泌型规范Wnt信号系统的抑制剂，通过与脂蛋白相关蛋白5和最近新发现的一种含kringle结构域的蛋白kremen形成三聚体复合物．这种复合物随即被细胞内吞，从而导致细胞表面Wnt受体脂蛋白相关蛋白5水平迅速下降，从而达到抑制Wnt信号通路的日地．通过对kremen和Ror2蛋白序列分析发现kremen和Ror2的胞外部分均含有一个结构上能与赖氨酸结合的kringle结构域．通过给怀孕母鼠注射一种赖氨酸类似物——氨甲环酸来研究kremen和Ror2的kringle结构域上的赖氨酸结合位点被占据对小鼠骨发育的作用．但是，研究结果表明AMCA组和对照组之间的骨密度并没有显著差异，揭示赖氨酸结合位点不参与骨的发育调控．     

硫酸钙富血小板血浆活性人工骨的制备及其理化性能研究

探讨了硫酸钙/富血小板血浆人工骨的制备及其理化性能。以100g/α-半水硫酸钙和富血小板血浆24mL(标准稠度)制备复合人工骨。经扫描电镜观察、X线晶体衍射分析、生物力学测试及体外降解实验,研究复合人工骨的组成结构、力学性能及降解特点。硫酸钙/富血小板血浆人工骨以5～10μm柱状晶体为主要骨架,其间布满直径约1μm的小片状或短柱状结晶,分布均匀,晶体间结合紧密。复合人工骨的抗压强度为38.5MPa。100%体外降解时间为41d。结果表明,硫酸钙/富血小板血浆人工骨可用作骨移植替代材料。     

L-丝氨酸与L-天门冬氨酸对羟基磷灰石仿生矿化的影响

根据仿生合成原理,以L-丝氨酸(L-Ser)和L-天门冬氨酸(L-Asp)为有机基质,通过气相扩散法制备碳-羟基磷灰石晶体,并在牙釉质片上进行仿生再矿化.通过场发射扫描电子显微镜(FESEM)、X射线衍射(XRD)和Fourier变换红外光谱(FTIR)表征产物结构,并考察不同浓度氨基酸对中间产物碳酸钙和最终产物羟基磷灰石粉体及牙釉质基质的影响.结果表明:氨基酸浓度对溶液中游离晶体粉末的形貌影响较大,由碳酸钙转化为羟基磷灰石,具有形貌遗传性,该影响未体现在牙釉质基质上;吸附在牙釉质基质片的氨基酸作为矿化成核位点,通过调节羟基磷灰石的晶体生长排列方向即可实现原位再矿化.     

多孔羟基磷灰石生物陶瓷的研究现状与进展

羟基磷灰石生物陶瓷与人体的无机组成和晶体结构相似 ,具有良好的生物活性和生物相容性 .多孔羟基磷灰石生物陶瓷 ,其相互连通的微孔有利于组织液的微循环并为羟基磷灰石深部的新生骨提供营养 ,促进纤维组织和新生骨的结合和生长 ,是一种性能优异的硬骨组织替代材料 .近年来 ,发明了一系列制备多孔羟基磷灰石陶瓷的方法 ,如有机泡沫浸渍法、添加造孔剂法、快速成型法等 .对于多孔羟基磷灰石生物陶瓷的研究 ,注重研究孔尺寸对多孔体与组织之间相互关系的影响 ,并逐步实现了对多孔体孔径的控制 .本文综述了多孔羟基磷灰石生物陶瓷的制备工艺、生物特性和发展趋势 .     

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.     

Bone indentation recovery time correlates with bond reforming time.

Despite centuries of work, dating back to Galileo, the molecular basis of bone's toughness and strength remains largely a mystery. A great deal is known about bone microsctructure and the microcracks that are precursors to its fracture, but little is known about the basic mechanism for dissipating the energy of an impact to keep the bone from fracturing. Bone is a nanocomposite of hydroxyapatite crystals and an organic matrix. Because rigid crystals such as the hydroxyapatite crystals cannot dissipate much energy, the organic matrix, which is mainly collagen, must be involved. A reduction in the number of collagen cross links has been associated with reduced bone strength and collagen is molecularly elongated ('pulled') when bovine tendon is strained. Using an atomic force microscope, a molecular mechanistic origin for the remarkable toughness of another biocomposite material, abalone nacre, has been found. Here we report that bone, like abalone nacre, contains polymers with 'sacrificial bonds' that both protect the polymer backbone and dissipate energy. The time needed for these sacrificial bonds to reform after pulling correlates with the time needed for bone to recover its toughness as measured by atomic force microscope indentation testing. We suggest that the sacrificial bonds found within or between collagen molecules may be partially responsible for the toughness of bone.     

The crystal structure of the photoprotein aequorin at 2.3 A resolution

Aequorin is a calcium-sensitive photoprotein originally obtained from the jellyfish Aequorea aequorea. Because it has a high sensitivity to calcium ions and is biologically harmless, aequorin is widely used as a probe to monitor intracellular levels of free calcium. The aequorin molecule contains four helix-loop-helix 'EF-hand' domains, of which three can bind calcium. The molecule also contains coelenterazine as its chromophoric ligand. When calcium is added, the protein complex decomposes into apoaequorin, coelenteramide and CO2, accompanied by the emission of light. Apoaequorin can be regenerated into active aequorin in the absence of calcium by incubation with coelenterazine, oxygen and a thiol agent. Cloning and expression of the complementary DNA for aequorin were first reported in 1985 (refs 2, 6), and growth of crystals of the recombinant protein has been described; however, techniques have only recently been developed to prepare recombinant aequorin of the highest purity, permitting a full crystallographic study. Here we report the structure of recombinant aequorin determined by X-ray crystallography. Aequorin is found to be a globular molecule containing a hydrophobic core cavity that accommodates the ligand coelenterazine-2-hydroperoxide. The structure shows protein components stabilizing the peroxide and suggests a mechanism by which calcium activation may occur.     

钛／羟基磷灰石涂层的电沉积过程及其结构特征

采用电沉积方法在钛金属基底上形成磷酸钙盐涂层,涂层经低温碱液处理获取羟基磷灰石涂层。研究了电流密度、主盐浓度、电解液温度、沉积电量、低温碱液处理对涂层表面形貌的影响,并用SEM、ZRD和IR对涂层的组成和结构进行分析。结果表明：电沉积磷酸钙盐涂层经低温碱液处理后得到纯的羟基磷灰石涂层,羟基磷灰石晶体呈针状结构;随电流密度、主盐浓度的增加,晶粒变粗,随电解液温度的升高,晶体发生变化,出现鳞片状结构;涂层质量随沉积电量的增加而增长。     

Crystal structure of the sodium-potassium pump

The Na+,K+-ATPase generates electrochemical gradients for sodium and potassium that are vital to animal cells, exchanging three sodium ions for two potassium ions across the plasma membrane during each cycle of ATP hydrolysis. Here we present the X-ray crystal structure at 3.5 A resolution of the pig renal Na+,K+-ATPase with two rubidium ions bound (as potassium congeners) in an occluded state in the transmembrane part of the alpha-subunit. Several of the residues forming the cavity for rubidium/potassium occlusion in the Na+,K+-ATPase are homologous to those binding calcium in the Ca2+-ATPase of sarco(endo)plasmic reticulum. The beta- and gamma-subunits specific to the Na+,K+-ATPase are associated with transmembrane helices alphaM7/alphaM10 and alphaM9, respectively. The gamma-subunit corresponds to a fragment of the V-type ATPase c subunit. The carboxy terminus of the alpha-subunit is contained within a pocket between transmembrane helices and seems to be a novel regulatory element controlling sodium affinity, possibly influenced by the membrane potential.     

Discussion on the mechanism of the calcium absorption in the human body

The present article discusses a new mechanism of calcium absorption in the human body. The mechanism is revealed as follows. First, after food is digested in the stomach, calcium ions (Ca2+) are released. The small intestine secretes amino acid or short peptide chain with small molecular weight automatically, which are called chelating agent; when the calcium ions from the stomach get to the small intestine, the reaction of the chelating agent with the calcium ions occurs, producing the neutral amino acid calcium chelate. Then, this kind of calcium chelate with small molecular weight is absorbed as a whole into the tissues of the small intestine. After being absorbed, in the cell the calcium chelate can break down its chelating bond automatically and decompose into the amino acid and calcium ion again. Finally, the calcium ion goes into blood through portal vein and is transferred to the organs and also deposits on the bone. The reason for the body抯 calcium insufficiency, which has no linear relation with the calcium intake amount, is the lack of the amino acid secreted by the small intestine. The main barrier that influences the calcium absorption is anion pollution. The calcium absorptivity of the body has nothing to do with the solubility of the calcium source out of the body. A new kind of calcium supplement agent——glycine calcium chelate——is synthesized, whose molecular weight is 206.06 (containing a molecular water). If the glycine calcium chelate is used to make calcium supplement agent, about 20 mg calcium element (converted from the glycine calcium chelate, the same below, no longer indicated) per day for one person, 50 mg at most, is enough to maintain the positive balance of calcium metabolism.     

Crystal structure of a bacterial homologue of Na+/Cl--dependent neurotransmitter transporters

Na+/Cl--dependent transporters terminate synaptic transmission by using electrochemical gradients to drive the uptake of neurotransmitters, including the biogenic amines, from the synapse to the cytoplasm of neurons and glia. These transporters are the targets of therapeutic and illicit compounds, and their dysfunction has been implicated in multiple diseases of the nervous system. Here we present the crystal structure of a bacterial homologue of these transporters from Aquifex aeolicus, in complex with its substrate, leucine, and two sodium ions. The protein core consists of the first ten of twelve transmembrane segments, with segments 1-5 related to 6-10 by a pseudo-two-fold axis in the membrane plane. Leucine and the sodium ions are bound within the protein core, halfway across the membrane bilayer, in an occluded site devoid of water. The leucine and ion binding sites are defined by partially unwound transmembrane helices, with main-chain atoms and helix dipoles having key roles in substrate and ion binding. The structure reveals the architecture of this important class of transporter, illuminates the determinants of substrate binding and ion selectivity, and defines the external and internal gates.