影响α-TCP和HA生物陶瓷表面形成类骨磷灰石的形貌、结构的因素研究

以磷酸三钙（α-TCP）和羟基磷灰石（HA）生物陶瓷为研究对象，采用X射线衍射（XRD）、扫描电子显微镜（SEM）研究αTCP和HA在模拟体液（SBF）及磷酸盐缓冲液（PBS）中浸泡后表面类骨磷灰石的生长和形成，考察初始溶液的离子成分和浓度及陶瓷溶解性能对形成类骨磷灰石的形貌、结构的影响.实验表明：在SBF及PBS中浸泡后，HA仅在SBF中有少量的类骨磷灰石颗粒生成；α-TCP在SBF中表面形成类骨磷灰石颗粒，而在PBS中形成片状的磷酸八钙（OCP），XRD和SEM结果分别从结构和形态学方面确认磷酸八钙在     

羟基磷灰石涂层修饰钛表面的蛋白质吸附及其对成骨细胞黏附和分化的作用(英文)

通过等离子体喷涂的方法在纯钛表面修饰羟基磷灰石涂层,可有效促进成骨分化并加速骨再生.生物材料表面的蛋白质吸附对后续细胞行为起着至关重要的作用,然而材料表面性能-蛋白质黏附-细胞行为之间的内在关系仍有待研究和阐明.本文检测了羟基磷灰石涂层(HA-Ti)和纯钛表面(Ti)吸附蛋白质层的性质,研究了蛋白质层对两种材料表面细胞黏附状态、死活细胞数和成骨分化的影响.相比Ti表面,HA-Ti表面纤维黏连蛋白(Fn)和血清蛋白吸附量较少,成骨细胞和干细胞黏附数量少;但HA-Ti表面对人成骨细胞BMP-2的吸附量稍大,诱导成骨分化明显.这说明HA涂层表面对蛋白质吸附性能的差异会导致细胞响应的不同,进而影响其生物学功能.     

纳米相陶瓷支架与人成骨细胞生物相容性的体外实验研究

研究了2种新型多孔纳米陶瓷支架材料(hydroxyapatite,HA和tricalcium phosphate,TCP)的生物相容性.从人4月龄胚胎骨膜组织中分离培养出骨膜来源的成骨细胞(periosteal-derived osteoblast,POB),采用人胚骨膜成骨细胞与2种陶瓷支架的体外复合培养,观察细胞在材料上的生长及生理功能表达情况.发现人POB在HA和TCP表面生长繁殖良好,并发挥成骨功能.材料对细胞的增殖还有一定的促进作用(p<0.05).实验表明: 2种新型多孔陶瓷支架材料均有良好的生物相容性.     

等离子体改性的4种口腔陶瓷的生物学性能研究

用等离子体活化改性口腔陶瓷材料BGC、HA、β-TCP和双相磷酸钙陶瓷,并用模拟体液(SBF)中类骨磷灰石的形成、体外成骨细胞培养、SEM、XPS、XRD等对其进行表征。结果发现:与活化改性前相比较,等离子体活化改性后的口腔陶瓷材料更有利于类骨磷灰石的形成,并能促进成骨细胞的增殖。活化机理是等离子体中丰富的高能、高活性的粒子轰击材料,使其被刻蚀和粗化,增加了表面的溶解性和提供了更多的活性位点,易使局域的钙、磷离子浓度达到过饱和,更有利于类骨磷灰石的成核和生长。表明等离子体改性提高了口腔陶瓷材料的活性。     

β－TCP／HA双相多孔磷灰石陶瓷成骨机制的研究

通过体外实验，确定了骨形成蛋白因子ＢＭＰ在多孔磷灰石陶瓷上的吸附位点，以及两者之间的吸附性能．在β－磷酸三钙／羟基磷灰石（β－ＴＣＰ／ＨＡ）双相多孔磷灰石陶瓷诱导成骨中，ＨＡ为宿主骨及β－ＴＣＰ降解提供钙、磷源的成骨机制．这些实验结果有助于增进对羟基磷灰石骨修复材料成骨作用的了解，并能较好地解释β－ＴＣＰ／ＨＡ双相磷灰石陶瓷在体内非骨环境中的诱导成骨作用．     

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。     

bHGA对体外培养成骨细胞活性的影响

为探讨12b-羟基-去-D-杰氏山竹子素A(bHGA)的体外潜在成骨活性,用bHGA处理MC3T3-E1成骨细胞,研究了其对细胞碱性磷酸酶活性、矿化、骨相关基因表达及早期成骨分化相关基因表达水平的影响.结果表明:bHGA处理细胞后,对Runx2 mRNA表达无显著影响,而对Osx mRNA的表达具有一定抑制作用.bHGA能同时促进骨钙蛋白基因OCN和Ⅰ型胶原Col 1 mRNA的表达,但抑制碱性磷酸酶mRNA的表达,其对骨桥蛋白基因OPN mRNA的表达没有显著影响.说明bHGA不利于骨早期分化,但它通过促进骨钙蛋白和I型胶原mRNA的表达而促进骨成熟.     

钛合金表面羟基磷灰石膜及组织相容性

在乙酸钙、磷酸二氢钙为主成分的弱酸性电解液中,以乙二胺四乙酸(EDTA)和柠檬酸作为络合剂,采用微弧氧化的方法,在钛合金表面成功地直接制备出含有羟基磷灰石(HA)的陶瓷层.随着电解液Ca/P的增加,膜层中HA含量升高,膜层表面孔洞分布更均匀,表面变粗糙.动物种植试验表明,陶瓷层覆盖植入体产生的炎症反应较弱,新的肌肉组织能够在膜层表面生长.     

粉体形态对网化HA陶瓷支架烧结和力学性能的影响:球形和针状HA粉体对比研究

以球形和针状的羟基磷灰石（HA）粉体作基料，采用有机泡沫浸渍法制备出高孔隙率的三维网状HA生物陶瓷支架，并研究粉体形态的不同对网化HA支架力学性能的影响，从而优化制备工艺以改善网化HA支架的力学性能，提高抗压强度．运用SEM、XRD、FTIR以及抗压测试考察了不同粉体对网状HA陶瓷支架的微观结构、相成分、烧结性能和力学性能的影响．结果表明：在相同的工艺条件下，球形粉体可以显著改善浆料的性能，影响浆料在支架上的涂覆量．因此球形粉体所制备的网状HA陶瓷支架的烧结性能和抗压强度明显优于针状粉体所制备的网状HA陶瓷支架，同时保持支架的贯通性和孔隙率．     

β—TCP／HA双相多孔磷灰石陶瓷成骨机制的研究

通过体外实验，确定了骨形成蛋白因子ＢＭＰ在多孔磷灰石陶瓷上的吸附位点，以及两者之间的吸附性能。在β－磷酸三钙／羟基磷灰石（β－ＴＣＰ／ＨＡ）双相多孔磷灰石陶瓷诱导成骨中，ＨＡ为宿主骨及β－ＴＣＰ降解提供钙、磷源有成骨机制。这些实验结果有助于增进对羟基磷灰石骨修复材料成骨作用的了解，并能较好地解释β－ＴＣＰ／ＨＡ双相磷灰石陶瓷在体内非骨环境中的诱导成骨作用。     

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 recognition mechanism of porcine osteocalcin from crystal structure

Osteocalcin is the most abundant noncollagenous protein in bone, and its concentration in serum is closely linked to bone metabolism and serves as a biological marker for the clinical assessment of bone disease. Although its precise mechanism of action is unclear, osteocalcin influences bone mineralization, in part through its ability to bind with high affinity to the mineral component of bone, hydroxyapatite. In addition to binding to hydroxyapatite, osteocalcin functions in cell signalling and the recruitment of osteoclasts and osteoblasts, which have active roles in bone resorption and deposition, respectively. Here we present the X-ray crystal structure of porcine osteocalcin at 2.0 A resolution, which reveals a negatively charged protein surface that coordinates five calcium ions in a spatial orientation that is complementary to calcium ions in a hydroxyapatite crystal lattice. On the basis of our findings, we propose a model of osteocalcin binding to hydroxyapatite and draw parallels with other proteins that engage crystal lattices.     

HMBA对人成骨肉瘤MG-63细胞形态结构和终末分化指标表达的影响

应用环六亚甲基双乙酰胺处理人成骨肉瘤MG-63细胞,观察MG-63细胞处理前后形态与超微结构及其相关终末分化指标的表达变化.实验结果显示,经5 mmol/L HMBA处理后,MG-63细胞体积增大,趋于扁平铺展状态,细胞大小较为一致,排列较为规则,细胞核形态规则,核质比例减小,核仁减少,核内异染色质减少,常染色质增多,细胞核内的线粒体和高尔基体较为发达,内质网数量增多,细胞表面的微绒毛减少,在成熟细胞中可见钙化糖原颗粒,细胞表面出现钙化小泡沉积,并且形成典型的骨结节.常规细胞化学和免疫细胞化学检测显示,HMBA处理后的细胞中Ⅰ型胶原纤维、骨钙素、骨粘素的表达显著增加,实验结果表明HMBA能够有效诱导人成骨肉瘤细胞的分化,并促进其终末分化指标的表达.     

Cytoskeleton reorganization and FAK phosphorylation are involved in lanthanum （Ⅲ）-promoted proliferation and differentiation in rat osteoblasts

The effect of lanthanum ion (La³⁺) on osteoblast function and cytoskeleton is assessed in vitro. Osteoblasts were isolated from Sprague-Dawley fetal neonatal rats. Cell proliferation and gene expression levels of cbfa-1, alkaline phosphatase (ALP), osteocalcin (OC), bone sialoprotein (BSP) and osteopontin (OPN) were examined by cell counting and RT-PCR. Cytoskeleton F-actin was stained with rhodamine-conjugated phalloidin and was visualized by a confocal microscope. As the results, 10⁻⁸-10⁻⁴ M La³⁺-induced osteoblast proliferation on day 2. Data from the RT-PCR assay revealed that 10⁻⁶ M La³⁺ up-regulated the expression levels of ALP, BSP, and cbfa-1 on day 4, while it enhanced the expressions of OC and OPN on day 21. The F-actin cytoskeleton was strengthened and reorganized under the exposure of La³⁺. In addition, the phosphorylation of focal adhesion kinase (FAK) was significantly promoted in 24 h evaluated by Western blot analysis. These findings indicate that La³⁺ promotes osteoblast activity through the phosphorylation of FAK and reorganization of the cytoskeleton.     

Osteogenic differentiation of mesenchymal stem cells promoted by overexpression of connective tissue growth factor

Objective: Large segmental bone defect repair remains a clinical and scientific challenge with increasing interest focusing on combining gene transfection with tissue engineering techniques. The aim of this study is to investigate the effect of connective tissue growth factor (CTGF) on the proliferation and osteogenic differentiation of the bone marrow mesenchymal stem cells (MSCs). Methods: A CTGF-expressing plasmid (pCTGF) was constructed and transfected into MSCs. Then expressions of bone morphogenesis-related genes, proliferation rate, alkaline phosphatase activity, and mineralization were examined to evaluate the osteogenic potential of the CTGF gene-modified MSCs. Results: Overexpression of CTGF was confirmed in pCTGF-MSCs. pCTGF transfection significantly enhanced the proliferation rates of pCTGF-MSCs (P<0.05). CTGF induced a 7.5-fold increase in cell migration over control (P<0.05). pCTGF transfection enhanced the expression of bone matrix proteins, such as bone sialoprotein, osteocalcin, and collagen type I in MSCs. The levels of alkaline phosphatase (ALP) activities of pCTGF-MSCs at the 1st and 2nd weeks were 4.0- and 3.0-fold higher than those of MSCs cultured in OS-medium, significantly higher than those of mock-MSCs and normal control MSCs (P<0.05). Overexpression of CTGF in MSCs enhanced the capability to form mineralized nodules. Conclusion: Overexpression of CTGF could improve the osteogenic differentiation ability of MSCs, and the CTGF gene-modified MSCs are potential as novel cell resources of bone tissue engineering.     

兔成骨细胞的体外培养及生物学行为观察

以一月龄幼兔的颅顶骨为材料 ,采用组织块法 ,进行细胞的原代培养并传代之 .结果显示 :(1)原代细胞具有成骨细胞的形态特征 ,经 6次传代后细胞形态变化不明显 ;(2 )细胞具有较强的碱性磷酸酶 (AL P)活性且稳中有升 ;(3)细胞培养液和细胞内均含有骨钙素 ;(4)连续培养 1个月后 ,成骨细胞有矿化结节形成 ;(5 )骨片经 4次贴壁仍有细胞爬出且细胞形态相同 .该结果表明所培养的细胞具有成骨细胞的生物学行为 ,为骨细胞生物学和骨组织工程的研究提供了一种客观而有效的手段     

大豆苷元对体外培养大鼠成骨细胞增殖与分化的影响

目的探讨大豆苷元对体外培养大鼠成骨细胞的增殖与分化能力的影响.方法从新生大鼠颅骨中分离培养大鼠成骨细胞,加入不同浓度的大豆苷元溶液进行培养,48,72 h后用MTT法测定成骨细胞的增殖,PNPP法测定碱性磷酸酶活性及RIA法测定骨钙素含量.结果不同浓度的大豆苷元作用48,72 h后,测定OD值均高于阴性对照组;成骨细胞在大豆苷元的作用下,与阴性对照相比,碱性磷酸酶活性显著增高;成骨细胞BGP含量显著增加.结论大豆苷元具有促进体外大鼠成骨细胞的增殖以及分化的作用.     

纳米羟基磷灰石陶瓷的制备及其力学性能的研究

利用两段式无压烧结的方法，制备了致密的纳米羟基磷灰石陶瓷，分别采用XRD，SEM和纳米压痕方法测试了纳米羟基磷灰石陶瓷的微观结构和力学性能.它与传统烧结方法制备的粗晶羟基磷灰石陶瓷的力学性能相比，随着晶粒尺寸的减小，纳米羟基磷灰石陶瓷的硬度提高了46.9%，弹性模量提高了23.4%，其中晶界增多和显微应力的产生是导致纳米羟基磷灰石陶瓷力学性能提高的主要原因.     

体外培养大鼠颅骨成骨细胞的生物学特性

应用机械和多步酶解的方法，从新生大鼠颅骨分离成骨细胞。偶氮染色法和钙钴法证明细胞的碱性磷酸酶活性呈阳性反应；Ｖｏｎ Ｋｏｓｓａ和钙分子探针Ｆｌｕｏ－３染色显示细胞节内有钙的沉积；电镜和倒置相差显微镜观察可见矿化结节及其形成过程；碱性磷酸酶性活性和骨钙素的动态分析提示这两种蛋白因子与成骨细胞分化与成熟的相关性。这一体外培养方法的建立，为骨的发育和成骨细胞生物学的研究提供了模型。