纳米羟基磷灰石/硫酸钙人工骨在模拟体液浸泡环境下的行为研究

目的 将纳米羟基磷灰石与半水硫酸钙组合构建一种可注射的人工骨材料,研究其在模拟体液环境中的降解与物质变化,并探讨该变化与材料成骨活性的关系.方法 采用模拟人体体液SBF9#浸泡两种晶体物质和不同成分组合的复合材料样品,测定失重率、钙磷元素释放速率,并对材料表面形态和晶体变化作扫描电镜观察和XRD分析.结果 纯羟基磷灰石在早期快速失重后保持稳定,其余材料均稳定降解,复合材料的HA含量越高,降解速率越快.纳米羟基磷灰石组的溶液Ca浓度始终较低;各组均有P元素释出,但随时间减少.XRD分析发现复合材料浸泡中CS峰下降,HA峰强度增高.扫描电镜观察到浸泡后复合材料表面呈现新的晶体形态.结论 CSD与HA在SBF环境下可能出现表面Ca、P的释放-吸附动态过程,材料的降解速度主要取决于CSD,可通过调节成分配比而加以控制.     

载药复合纳米人工骨的注射、凝固、抗稀散性及药物缓释放的性能研究

目的 考察纳米羟基磷灰石(n-HA)的载药情况,并复合α-半水硫酸钙(CSH)构建复合人工骨材料,比较载药与未载药的复合人工骨的各方面性能,以及对复合材料的药物缓释放情况进行考察.方法 用离子交换法把阿莫西林载入n-HA中,与未载药的n-HA用UV-vis和TEM进行对比检测.再分别以质量比1:1与CSH复合,并对两组复合人工骨的各方面性能进行对比考察.把复合材料浸泡于生理盐水,每天更新浸泡液,测试骨水泥中药物的释放效果.结果 n-HA可有效地运载入阿莫西林.载药组的注射性能优于未载药组,凝固时间、抗稀散性相差不大.药物在浸泡数天内持续释放.结论 n-HA可以作为药物缓释放载体,以质量比1:1与CSH构建复合人工骨材料,其注射性能、凝固时间、抗稀散性均能符合临床要求.     

纳米羟基磷灰石/硫酸钙复合人工骨的生物安全性研究

目的研制纳米羟基磷灰石/半水硫酸钙(n-HA/CSH)复合型人工骨,并对其进行体内、外生物安全性测试。方法对n-HA/CSH人工骨进行急性全身毒性试验、皮内刺激试验、致敏试验、MTT细胞毒性试验和遗传毒性实验(Ames试验)并与对照组比较。结果人工骨浸取液静脉及腹腔注射后不引起小鼠呼吸、进食改变或死亡,体重稳定。家兔皮内注射72小时后仅出现红斑或微弱水肿,豚鼠皮内注射后未出现过敏反应。MTT细胞毒性试验显示含HA10%、20%、40%人工骨及纯n-HA、CSH的细胞增殖率均在77%以上,细胞毒性均为0～1级,Ames试验表明含HA40%人工骨的不同浓度生理盐水浸取液引起鼠伤寒沙门氏菌回复突变数均不超过阴性对照组的2倍。结论n-HA/CSH复合材料不引起全身毒性反应、皮内刺激反应和急性过敏反应。且无MTT细胞毒性,细胞相容性良好。同时,复合材料的生理盐水浸取液不引起鼠伤寒沙门氏菌回复突变数增加。     

β-磷酸三钙/α-半水硫酸钙复合人工骨体外降解速度可与成骨一致

背景：骨修复材料的理想降解速度应该与新骨形成速度相匹配,逐渐被新生骨逐渐爬行替代。 目的：探讨β-磷酸三钙/α-半水硫酸钙复合人工骨的体外降解速度。 方法：将β-磷酸三钙/α-半水硫酸钙复合人工骨、β-磷酸三钙、α-半水硫酸钙试件置于PBS模拟体液中,测量不同浸泡时间的pH变化、试件降解率和压缩强度变化。 结果与结论：在模拟体液中复合人工骨和α-半水硫酸钙的pH值随时间延长逐渐降低,而β-磷酸三钙的pH值变化不大,4周后复合人工骨的pH值稳定在5.6左右。PBS模拟体液浸泡过程中,复合人工骨与α-半水硫酸钙的质量和压缩强度均随时间延长而不断降低,而β-磷酸三钙的质量随时间的变化降低较小。说明β-磷酸三钙/α-半水硫酸钙复合人工骨通过物理溶解而逐渐降解,其体外降解率介于β-磷酸三钙和α-半水硫酸钙之间。     

原位法制备羟基磷灰石/聚乳酸纳米复合材料的体外生物学研究

通过羟基磷灰石/聚乳酸(HA/PLLA)纳米复合材料在模拟体液(SBF)中的浸泡实验评价该材料的生物学性能。测试发现HA/PLLA纳米复合材料在浸泡过程中SBF的pH值呈现下降趋势,HA的存在缓冲了PLLA的酸性;复合材料表面有类骨磷灰石层沉积,并有"蚕茧状"类骨磷灰石颗粒和夹有短棒状晶体的片状晶体簇生成;同时复合材料的降解导致表面形成大量蜂巢状多孔。因此原位法制备的HA/PLLA纳米复合材料具有较好的生物活性和可降解性。     

羟基磷灰石/聚乳酸复合人工骨修复材料的研究进展

近年来,羟基磷灰石/聚乳酸复合生物材料用做人工骨修复材料得到了广泛的研究。本文综述了近年来羟基磷灰石/聚乳酸(HA/PLA)复合人工骨修复材料的各种制备方法及其性能影响因素的研究进展。     

SrHA/PMMA复合骨水泥的制备、理化性能及细胞相容性研究

目的结合PMMA和锶羟基磷灰石(Sr HA)各自的优势,制备出兼具高的力学强度、合适的固化时间、较低的热释放、生物活性和骨整合性能的Sr HA/PMMA复合骨水泥,并系统性地研究Sr HA的引入对复合骨水泥的体外固化性能、力学强度和生物学性能的影响。方法将水热合成法制备的锶羟基磷灰石引入PMMA基体,制备Sr HA/PMMA复合骨水泥。系统性地对Sr HA/PMMA复合骨水泥的力学强度、固化时间、热释放、生物活性进行研究。将复合骨水泥和细胞共培养,利用MTT法、扫描电镜等研究Sr HA/PMMA复合骨水泥的细胞毒性,粘附和增殖。结果结果表明,与纯的PMMA骨水泥(对照组)相比,Sr HA/PMMA复合骨水泥的固化热释放明显降低(约80~84℃),同时又维持了合适的固化时间(8~11分钟)和较好的力学性能(抗压强度为90MPa左右)。Sr HA的引入,不仅赋予了复合骨水泥生物活性,也显著地改善了其细胞/材料的相互作用。浸泡在SBF后,Sr HA/PMMA复合骨水泥显示出更好的体外矿化性能。与成骨细胞MC3T3-E1共培养后,表面沉积的羟基磷灰石能够更好的促进细胞的粘附和爬行。结论兼具优异的理化性能和生物活性的Sr HA/PMMA复合骨水泥,有着广阔的骨科微创修复应用前景。     

人工颈椎间盘不同涂层影响骨髓间充质干细胞的黏附及分化

背景：钛合金人工颈椎间盘具有良好的生物相容性,但钛合金存在生物活性差、与骨结合强度低、在生理环境下易造成金属离子释放等问题。 目的：观察国产人工颈椎间盘钛合金终板不同涂层对大鼠骨髓间充质干细胞黏附、分化的影响。 方法：将第3代Wistar大鼠骨髓间充质干细胞,接种于含羟基磷灰石涂层、钛粉复合羟基磷灰石涂层的钛合金板及裸钛合金板的24孔板内,培养的第24,48小时后分别终止培养,扫描电镜观察细胞生长情况;接种24 h后加成骨诱导剂进行诱导,并于第3,5,7天各收集细胞裂解后离心的上清,检测碱性磷酸酶表达。 结果与结论：骨髓间充质干细胞与材料复合培养48 h后,羟基磷灰石组和钛粉复合羟基磷灰石组表面的细胞呈多角形,并伸出细长伪足伸入到材料的微孔内,与材料表面紧密黏附;而裸钛合金组骨髓间充质干细胞分化较差且黏附率低。随时间延长,各组碱性磷酸酶表达均增加,羟基磷灰石组和钛粉复合羟基磷灰石组各时间点的碱性磷酸酶表达均较裸钛合金组明显增高(P < 0.05)。表明羟基磷灰石、钛粉复合羟基磷灰石涂层可有促进大鼠骨髓间充质干细胞的黏附和分化。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

纳米微球负载抗生素/羟基磷灰石复合支架的缓释性能及治疗感染性骨缺损的研究

目的 评价新型纳米微球负载抗生素/羟基磷灰石复合支架材料的体外缓释性能及治疗感染性骨缺损的疗效。方法 运用HPLC法检测不同时间点药物的体外释放量。建立兔胫骨感染性骨缺损动物模型24只,共分为4组：a组动物仅单纯清创;b组植入1 mg Lev/PMMA;c组植入n-HA/PU;d组植入1 mg Lev/n-HA/PU。在植入材料术后6周、12周观察放射学、组织病理学,Micro CT评价新骨生成。评价新型复合材料控制感染、诱导成骨能力、治疗感染性骨缺损的效果。结果 1 mg Lev/n-HA/PU组比1 mg Lev/PMMA组释放的抗生素更多（P<0.05）,缓释性能表现更好。单纯清创组X线表现出局部骨破坏、死骨形成。1 mg Lev/n-HA/PU组无明显骨破坏。Micro CT发现1 mg Lev/n-HA/PU组材料周围新生骨小梁数量与其余三组比较,差异有统计学意义（P<0.05）,结论 新型载抗生素复合支架材料具有良好的缓释性能、抗感染能力、骨诱导能力,可有效治疗胫骨感染性骨缺损。     

羟基磷灰石骨水泥中碳酸根存在的意义及其对溶解度的影响

目的探讨羟基磷灰石骨水泥中碳酸根存在的意义及其对溶解度产生的影响。方法制备碳酸化羟基磷灰石骨水泥，定量分析其固化产物晶体中的碳酸根结构。以磷酸钙骨水泥为对照组，分别预制直径为8mm、高为12mm的圆柱体试样。经模拟体液浸泡后，定期观察、测量每一试样的失重率，对照分析其体外溶解性的差别。结果碳酸化羟基磷灰石骨水泥固化产物的晶体中含有质量浓度5．6％的碳酸根，与自然骨的矿物成分更接近。随着时间的改变，材料在模拟体液中逐渐由白色转为微黄色，棱角逐渐变钝。体视显微镜下观察发现材料表面出现点、坑、片状溶蚀，碳酸化羟基磷灰石骨水泥较磷酸钙骨水泥更明显。失重率的检测表明，酸化羟基磷灰石骨水泥的失重率改变明显高于磷酸钙骨水泥。结论羟基磷灰石骨水泥中碳酸根的存在可增加其溶解度，加快降解速度。     

Influence of carboxymethyl chitin on stability and biocompatibility of 3D nanohydroxyapatite/gelatin/carboxymethyl chitin composite for bone tissue engineering

A novel three-dimensional (3D) scaffold has been developed from the unique combination of nanohydroxyapatite/gelatin/carboxymethyl chitin (n-HA/gel/CMC) for bone tissue engineering by using the solvent-casting method combined with vapor-phase crosslinking and freeze-drying. The surface morphology and physiochemical properties of the scaffold were investigated by dissolvability test, infrared absorption spectra (IR), X-ray diffraction (XRD), transmission electron microscope (TEM), scanning electron microscope (SEM), mechanical testing, and soaking in simulated body fluid (SBF). An optimized (composition and processing parameters) ratio of n-HA:gel:CMC (1:2:1), exhibited ideal porous structure with regular interconnected pores (75-250 μm) and higher mechanical strength. Result suggested that the divalent (Ca(++)), carboxyl (COO(-)), amino (NH4(+)), and phosphate (PO4(3-)) groups created favorable ionic interactions which facilitated structural stability and integrity of the composite scaffold. The SBF soaking experiment confirmed the apatite nucleation ability, induced by CMC incorporation. Furthermore, hemocompatibility (hemolysis, platelet adhesion, and protein adsorption) and biocompatibility with MG63 osteoblast cells (MTT assay, cell morphology, and confocal studies from within the 3D scaffold) indicated that the structural and dimensional stability of composite scaffold provided an optimal mechanosensory environment for enhancement of cell adhesion, proliferation, and network formation. The n-HA/gel/CMC composite, therefore, may serve as a promising composite scaffold for guided bone regeneration.     

Employing the cyclophosphate to accelerate the degradation of nano-hydroxyapatite/poly(amino acid) (n-HA/PAA) composite materials

Owing to the good degradability and biocompatibility of polyphosphoesters (PPEs), the aim of the current study was to investigate a novel degradable composite of nano-hydroxyapatite/poly(amino acid) (n-HA/PAA) with cyclophosphate (CPE) via in situ melting polymerization to improve the degradation of n-HA/PAA. The structure of each composite was characterized via Fourier transform infrared spectroscopy, X-ray diffraction, and X-ray photoelectron spectroscopy. The degradation properties were studied in terms of the weight loss and pH in a phosphate-buffered saline (PBS) solution, while the surface morphology was examined using a scanning electron microscope-energy dispersive spectrometer (SEM-EDS) after soaking the surface in simulated body fluid (SBF). The cell proliferation, cell adhesion, and alkaline phosphatase (ALP) activity were used for the analysis of cytocompatibility. The weight loss results showed that the n-HA/PAA composite was 9.98 wt%, weighed after soaking in the PBS solution for 12 weeks, whereas the nano-hydroxyapatite/polyphosphoester-amino acid (n-HA/PPE-AA) composite was 46.94 wt%. The pH of the composites was in a suitable range between 6.64 to 7.06 and finally stabilized at 7.39. The SEM and EDS results revealed the formation of an apatite-like layer on the surface of the n-HA/PPE-AA composites after soaking in SBF for one week. The cell counting Kit 8 (CCK-8) assay of the cell culture in the leaching liquid of the n-HA/PPE-AA composites exhibited non-cytotoxicity and high-proliferation, and the cell adhesion showed the well spreading and normal phenotype extension of the cells on the n-HA/PPE-AA composites surface. Concurrently, the co-culture results of the composites and cells confirmed that the n-HA/PPE-AA composites exhibited a higher ALP activity. In summary, the results demonstrated that the n-HA/PPE-AA composites had a controllable degradation property, good bioactivity, and cytocompatibility.     

Preparation and characterization of nano-hydroxyapatite/chitosan/konjac glucomannan composite

Nano-hydroxyapatite (n-HA)/chitosan (CS)/konjac glucomannan (KGM) composite was prepared by coprecipitation method and investigated by thermal gravitivity/differentiate thermal analysis, Fourier transform infrared spectroscopy, X-ray diffraction, inductively coupled plasma emission spectroscopy, scanning electron microscopy, and energy dispersive X-ray analyzer. The analyses showed that the three phases of n-HA, CS, and KGM combined closely to each other. Further, in vitro tests were conducted to investigate the degradation and bioactivity of the composite. During immersion in simulated body fluid (SBF), pores appeared and a new substance containing Ca and P formed on the surface of the composite. Also, the concentration of Ca and P in SBF changed and weight loss of the composite was observed during time. The composite revealed a high degradation in SBF. Evidently, the new composite has a potential to be used as a carrier of implantable drug delivery system. The biodegradation rate and route could be different from CS and KGM, which will provide an opportunity to control the degradation rate or drug releasing rate by simply adjusting the ratio of CS and KGM.     

玉米蛋白3-D多孔支架材料仿生矿化的初步研究

为了提高玉米蛋白的骨诱导性和骨结合性,以促进其在骨组织工程领域中的应用,将玉米蛋白多孔支架材料浸泡于5倍模拟体液(5×SBF)中,尝试在不同的条件下对玉米蛋白多孔支架材料表面进行仿生矿化。通过场发射扫描电镜(SEM)观察仿生矿化后多孔支架材料各个表面的形貌,能谱(EDS)计算出钙磷比(Ca/P)比。在5倍模拟体液中浸泡后,材料各表面均形成了分布和尺寸相对均匀的微米级(2~10 um)颗粒。其钙磷比接近羟基磷灰石,可以认为获得了比较理想的玉米蛋白-羟基磷灰石多孔支架复合材料。     

Fabrication and characterization of bioactive wollastonite/PHBV composite scaffolds

Composite scaffolds of polyhydroxybutyrate-polyhydroxyvalerate (PHBV) with bioactive wollastonite were fabricated by a compression moulding, thermal processing, and salt particulate leaching method. Structure and mechanical properties of the scaffolds were determined. The bioactivity of the composites was evaluated by soaking in a simulated body fluid (SBF), and the formation of the hydroxyapatite (HAp) layer was determined by Scanning Electron Microscope (SEM) and Energy-Dispersive Spectrometer (EDS). The results showed that the wollastonite/PHBV composites were bioactive as it induced the formation of HAp on the surface of the composite scaffolds after soaking in SBF for 14 days. In addition, the measurements of the water contact angles suggested that incorporation of wollastonite into PHBV could improve the hydrophilicity of the composites and the enhancement was dependent on the wollastonite content. Furthermore, the pH and ion concentration changes of SBF solutions with composite scaffolds showed that the composites released Ca and Si ions, which could neutralize the acidic by-products of the PHBV and stabilize the pH of the SBF solutions between 7.2 and 7.8 within a 3-week soaking period. All of these results suggest that the incorporation of wollastonite was a useful approach to obtain composite scaffolds with improved properties.     

中空羟基磷灰石复合rhBMP-2修复骨缺损过程的再血管化研究

目的　探讨和观察中空羟基磷灰石复合rhBMP-2在骨缺损修复过程的再血管化。  方法　将48只成年的新西兰雄性大白兔制作成桡骨骨缺损模型,随机分3组,各组分别植入以下材料：中空HA/ rhBMP-2复合人工骨、单纯中空HA人工骨、单纯rhBMP-2。植入后于4、8、12、16周分别注射99mTc-MDP进行放射性核素骨显像并监测骨缺损修复过程中再血管化情况,同时进行大体、X线、组织学观察。  结果　术后各时间段,中空HA/ rhBMP-2复合人工骨组在X线及放射性核素聚集强度明显高于单纯中空HA人工骨组(P<0.05) ,表现为成骨代谢活跃及早期的再血管化能力。  结论　中空HA/ rhBMP-2复合人工骨具有良好的骨缺损修复能力,成骨活性持久,再血管化能力强,有望成为一种理想的骨缺损修复材料。     

In vitro testing of calcium phosphate (HA, TCP, and biphasic HA-TCP) whiskers

Calcium phosphate [single-phase hydroxyapatite (HA, Ca(10)(PO(4))(6)(OH)(2)), single-phase tricalcium phosphate (beta-TCP, Ca(3)(PO(4))(2)), and biphasic HA-TCP] whiskers were formed by using a novel microwave-assisted molten salt mediated process. Aqueous solutions containing NaNO(3), HNO(3), Ca(NO(3))(2) x 4H(2)O, and KH(2)PO(4) (with or without urea) were used as starting reagents. These solutions were irradiated in a household microwave oven for 5 min. As-recovered precursors were then simply stirred in water at room temperature for 1 h to obtain the whiskers of the desired calcium phosphate (CaP) bioceramics. These whiskers were evaluated, respectively, in vitro by (1) soaking those in synthetic body fluid (SBF) solutions at 37 degrees C for one week, and (2) performing cell attachment and total protein assay tests on the neat whiskers by using a mouse osteoblast cell line (7F2). beta-TCP, HA, and HA-TCP biphasic whiskers were all found to possess apatite-inducing ability when soaked in SBF. SBF-soaked whiskers were found to have BET surface areas ranging from 45 to 112 m(2)/g. Although the osteoblast viability and protein concentrations were found to be the highest on the neat HA whiskers, cells were attached and proliferated on all the whiskers.     

新型有序纳米介孔生物活性玻璃在模拟体液中的表面生物活性研究

目的评价新型行序纳米介孔生物活性玻璃（80SMBG）的体外生物活性及其对体液环境的影响。方法以Novabone为对照,利用模拟人体体液体外浸泡实验（SBF）的方法和扫描电镜（SEM）、等离子发射光谱仪（ICP）pH仪等技术,比较80SMBG与Novabone体外生成类似于天然骨中无机矿物的羟基磷灰石（HA）的能力,分析SBF中各离子浓度、pH值的变化情况。结果80SMBG在SBF中浸泡4h表面即有羟基磷灰石（HA）生成,浸泡1～4hSBF中Ca、P、Si的离子浓度迅速达到峰值,8h后逐渐降低直至稳定,pH仅在7．40～7．65范围内变化。结论80SMBG的体外生物活性更高,对体液微环境的影响更轻微,作为骨组织修复替代材料或骨组织工程支架材料具有较高的研究和应用价值。