A study of apatite formation on natural nano-hydroxyapatite/ chitosan composite in simulated body fluid

This study is focused on the ability of apatite formation on the surface of nano-hydroxyapatite (HA)/chitosan (CH) composite in simulated body fluid (SBF) in vitro. At first, natural nano-HA was prepared according to a wet-balling method and the composite was prepared by combining the natural nano-hydroxyapatite and chitosan, and then in vitro biomineralization test of natural nano-HA/CH composite was carried out in standard SBF. Subsequently, the quantity of the weight of the particles formed on the composite surface in SBF was measured by analytical balance, and the morphology change on the surface of the composite was observed by a scanning electron microscope (SEM). Lastly, a Fourier transform infrared spectroscope (FTIR) was used to investigate the chemical components of the particles formed on the natural nano-HA/CH composite surface in SBF. The result of quantity assessment shows that the weight of the composite increased with the increase of soaking time. The SEM image shows that the particles were gradually formed on natural nano-HA/CH composite surface, and the FTIR spectrum of the particles on composite surface confirms that these particles were carbonate apatite. This study indicates that the nano-HA/CH composite has a good ability for apatite formation in SBF, which predicts the bone-inducing ability of natural nano-HA/CH composite in vivo.     

Hydrolysis of monetite/chitosan composites in α-MEM and SBF solutions

There are two objectives of this work. The first objective is to study the hydrolysis behavior of monetite cements formed in the presence and absence of the chitosan in cell culture media (α-MEM) and simulated body fluid (SBF) solutions at 37°C. During hydrolysis, monetite transformed to carbonated apatite. Therefore, the second objective is to examine how addition of chitosan affects on the formation of carbonated apatite phases. The changes in the phase structure of monetite after hydrolysis reactions were characterized using XRD, FTIR and SEM. Pure monetite and monetite/chitosan composite were soaked in α-MEM and SBF solution for 4 and 7 days. In α-MEM solution, the monetite particles started to transform into carbonated apatite with a slow rate. However, in SBF, the rate of monetite transformation to carbonated apatite was more rapid. The presence of the chitosan had no significant effect on the precipitation of carbonated apatite on the monetite particles.     

In vitro evaluation of hydroxyapatite reinforced polyhydroxybutyrate composite

Particulate hydroxyapatite (HA) was incorporated into polyhydroxybutyrate (PHB) to form a bioactive and biodegradable composite for applications in hard tissue replacement and regeneration. HA/PHB composite containing 10, 20, and 30 vol.% of HA was made for in vitro evaluation. In vitro studies were conducted using an acellular simulated body fluid (SBF). Composite specimens were immersed in SBF at 37 °C for various periods of time prior to surface analysis and mechanical testing. Results obtained from scanning electron microscopic (SEM) examination, thin film X-ray diffraction (TF-XRD) analysis, and Fourier transform infrared (FTIR) spectroscopy showed that a layer of bone-like apatite formed within a short period on HA/PHB composite after its immersion in SBF, demonstrating high in vitro bioactivity of the composite. The bioactivity and mechanical properties of the composite could be changed by varying the amount of HA in the composite. Dynamic mechanical analysis (DMA) revealed that the storage modulus (E′) of HA/PHB composite increased initially with immersion time in SBF, due to apatite formation on composite surface and decreased after prolonged immersion in SBF, indicating degradation of the composite in a simulated body environment. HA/PHB composite thus has the potential for its intended applications.     

可注射nano HA/PAG/CS复合材料的制备与表征

通过原位聚合法制备了可注射纳米羟基磷灰石/天门冬氨酸-谷氨酸共聚物/硫酸钙复合材料(HA/PAG/CS), 采用FTIR、XRD、SEM对复合材料的组成结构、表面形貌及力学性能进行了表征, 研究了复合材料在模拟体液(SBF)中的降解性能。结果显示: 复合材料无机相羟基磷灰石、硫酸钙与有机相天门冬氨酸-谷氨酸共聚物之间存在化学相互作用, 具有良好的抗压强度; 7周后, 复合材料在SBF中完全降解, 降解方式为表面降解; 在降解过程中, 浸泡液的pH值在6.4~7.4之间变化; 复合材料在SBF中浸泡后, 其表面能够沉积磷灰石, 表明复合材料具有良好的生物活性, 有利于植入体与骨组织形成良好的界面结合。     

<Emphasis Type="Italic">In vitro</Emphasis> bioactivity studies of larnite and larnite/chitin composites prepared from biowaste for biomedical applications

Larnite (Ca₂SiO₄) was synthesized by the sol–gel combustion process by using raw eggshell powder as a calcium source and urea as a fuel. The main focus of this work is to convert biowaste into a biomedical material at a low-processing temperature. X-ray diffraction (XRD) pattern confirms the phase purity of the larnite and Fourier transform infrared (FTIR) spectra confirms the presence of characteristic functional groups of larnite. Scanning electron microscopy (SEM) image shows agglomerated particles with cauliflower-like morphology and energy dispersive X-ray spectroscopy (EDX) confirms the presence of the stoichiometric ratio of required elements. Atomic force microscope (AFM) images reveal the presence of pores on the surface of spherical particles. Larnite/chitin composites were fabricated into scaffold with different ratios of bioceramic to biopolymer (70:30, 80:20) to investigate the influence of the polymer content on the apatite formation ability in simulated body fluid (SBF) medium. XRD pattern and FTIR spectra of the scaffold immersed in SBF shows apatite deposition within 5 days. The deposition of hydroxyapatite (HAP) on the scaffold surface increases with the increase in polymer content of the composite.     

表面改性对Cf/HA-PMMA混杂生物复合材料的结构及性能的影响

采用原位合成与溶液共混相结合的方法,制备了短切碳纤维(C_f) 增强纳米羟基磷灰石(HA)-聚甲基丙烯酸甲酯(PMMA)生物复合材料。重点研究了短切碳纤维和纳米HA粒子表面改性前后对C_f/HA-PMMA复合材料微观结构和力学性能的影响。采用XRD、FTIR、XPS和SEM等对纳米HA粒子、碳纤维和复合材料的组成结构及断面的微观形貌等进行测试和表征,使用万能材料试验机测试其弯曲、压缩性能。结果表明：经表面氧化的碳纤维和用卵磷脂改性后的纳米HA与PMMA基体的界面结合性明显得到改善;采用卵磷脂表面改性后的纳米HA及表面预氧化后的碳纤维制备的C_f/HA-PMMA复合材料的弯曲性能得到显著提高,与采用未表面改性纳米HA和未表面氧化碳纤维所制备的C_f/HA-PMMA复合材料相比,弯曲、压缩强度和弹性模量分别提高1.6倍、2倍和4.3倍。     

纳米羟基磷灰石/聚碳酸酯复合生物材料Ⅱ:体外生物活性

研究了纳米羟基磷灰石/聚碳酸酯(n-HA/ PC)生物复合材料在模拟体液(SBF)中的表面变化,并用傅里叶红外光谱(FTIR) 、X射线衍射仪(XRD)和扫描电子显微镜(SEM)对材料的表面变化进行了分析。结果表明,n-HA/PC生物复合材料在模拟体液(SBF)中浸泡后,表面会沉积碳酸化羟基磷灰石(CHA),随着浸泡时间的延长,沉积层变厚,CHA晶体形貌变得规整。对n-HA/PC复合材料进行了细胞实验,通过四唑盐(MTT)检测和扫描电镜观察,表明n-HA/PC复合材料无细胞毒性,细胞形态正常,是一种有应用前景的可承力骨修复替代材料。      

Preparation and properties of a novel bone repair composite: nano-hydroxyapatite/chitosan/carboxymethyl cellulose

Nano-hydroxyapatite/chitosan/carboxymethyl cellulose (n-HA/CS/CMC) composites with weight ratios of 70/10/20, 70/15/15 and 70/20/10 were prepared through a co-solution method. The properties of the composites were characterized by means of burn-out test, IR, XRD, TEM and universal material testing machine. The degradation and bioactivity were also investigated by in vitro test in a simulated body fluid (SBF) for 8 weeks. The results showed that n-HA particles were dispersed uniformly in organic phase, and strong chemical interactions formed among the three phases. Moreover, the composites were similar to natural bone in morphology and size. In addition, the compressive strength was improved compared with n-HA/CS composite. The biodegradation rate was controllable by altering weight ratio of the CS/CMC. Meanwhile, the composites could induce apatite particles to deposit in SBF. All the above results indicate that the novel composites of n-HA/CS/CMC have a promising prospect used for bone repair materials in view of the good mechanical property, adjustable biodegradation rate and bioactivity in SBF. Additionally, the study would provide a good guide to exploit clinical application of natural cellulose.     

一种新型仿生骨组织工程支架的制备与表征

利用改性生物玻璃粉体和胶原、透明质酸钠、磷酸丝氨酸等天然生物分子复合制备仿生型三维多孔骨组织工程支架材料,利用体外模拟实验结合SEM、FTIR、XRD 等测试方法对材料的显微结构、生物矿化性能进行了综合研究,研究表明该材料具有良好的孔隙结构,在模拟生理溶液(SBF)中反应24h即可在支架表面形成碳酸羟基磷灰石(HCA).     

In vitro study of nano-sized zinc doped bioactive glass

Surface reactivity in physiological fluid has been linked to bioactivity of a material. Past research has shown that bioactive glass containing zinc has the potential in bone regeneration field due to its enhanced bioactivity. However, results from literature are always contradictory. Therefore, in this study, surface reactivity of bioactive glass containing zinc was evaluated through the study of morphology and composition of apatite layer formed after immersion in simulated body fluid (SBF). Nano-sized bioactive glass with 5 and 10 mol% zinc were synthesized through quick alkali sol–gel method. The synthesized Zn–bioglass was characterized using field emission scanning electron microscope (FESEM), energy dispersive X-ray spectrometer (EDX), X-ray diffractometer (XRD) and Fourier transform infrared spectrometer (FTIR). Samples after SBF immersion were characterized using scanning electron microscope (SEM) and EDX. Morphological study through SEM showed the formation of spherical apatite particles with Ca/P ratio closer to 1.67 on the surface of 5 mol% Zn–bioglass. Whereas, the 10 mol% Zn–bioglass samples induced the formation of flake-like structure of calcite in addition to the spherical apatite particles with much higher Ca/P ratio. Our results suggest that the higher Zn content increases the bioactivity through the formation of bone-bonding calcite as well as the spherical apatite particles.     

Preparation and in vitro bioactivity of poly(d,l-lactide) composite containing hydroxyapatite nanocrystals

The nano-sized hydroxyapatite (n-HA) was incorporated into poly(d,l-Lactide) (PDLLA) to form a bioactive and biodegradable composite for application in hard tissue replacement and regeneration. Thin film of PDLLA composite containing 20 mass% of n-HA fillers was successfully developed through integration of solvent co-blending and hot pressing techniques. firstly, n-HA and PDLLA were chemically synthesized, respectively, then mixed together and homogeneously dispersed in N,N-dimethyl formamide(DMF) solvent, finally, the dried blended hybrid containing PDLLA matrix and n-HA fillers was put into the mould and compacted by hot-pressing machine under 8 MPa pressure at 110 °C for 15 min. In vitro studies were conducted using the simulated body fluid(SBF). Composite specimens were soaked in SBF from 1 day to 21 days prior to surface analysis. Results obtained from scanning electron microscopy(SEM) examination, Energy dispersive X-ray detector(EDX) analysis and X-ray diffraction (XRD) analysis showed that a layer of non-stoichiometric apatite formed within 7 days on HA/PDLLA composite surface after its immersion in SBF, demonstrating moderate in vitro bioactivity of n-HA/PDLLA composite, though a moderate rate of apatite formation in SBF was found on initial stage of immersion periods for n-HA/PDLLA composite, compared to the other biomaterial composite. This type of composite film exhibited certain desirable bioactive characteristics, and they are promising bone candidates to develop novel bioactive composites for biomedical application.     

纳米HA/PA6复合材料的体外生物活性

研究了PA6和纳米HA/PA6复合材料在模拟体液(SBF)中的行为变化,用IR,XRD,SEM和EDS等手段对材料的表面变化进行了分析,讨论了PA6和纳米HA/PA6复合材料的稳定性、亲水性和生物活性。结果表明:在SBF中PA6的吸水率大概在6%左右,纳米HA/PA6复合材料的吸水率有少量下降,PA6和纳米HA/PA6复合材料出现一定的溶解和降解。在SBF中,PA6表面形成Ca,P化合物中的Ca/P比例为1.12,与HA的理论值1.67有一定的差别;HA/PA6复合材料在其表面形成了HA沉积物和碳酸取代的磷灰石沉积物,Ca/P逐步变化为1.67,表现出较好的生物活性。复合材料表面沉积的HA和原来合成的HA具有相近的结晶形貌,该复合材料可作为优良的骨修复填充材料和组织工程支架材料。      

磷灰石包覆金红石型TiO2纳米粉体的制备及表征

将金红石型TiO₂纳米粉体浸泡在模拟体液中, 于37℃经过不同时间, 制备出磷灰石包覆金红石型TiO₂纳米粉体. 用XRD、SEM、TEM、EDX、FTIR、ICP AES和BET方法对复合粉体进行了表征, XRD结果表明,磷灰石的含量可随着金红石型TiO2在模拟体液中浸泡时间的延长而增加. FTIR结果中显示了磷灰石的O-H和PO₄^3-吸收峰, 说明复合粉体中有磷灰石存在. ICP-AES结果表明溶液中Ca和P浓度随浸泡时间延长而下降, 表明时间延长后更多的Ca和P被消耗. TEM和EDX结果证明了金红石型TiO₂表面有磷灰石存在, HRTEM结果显示磷灰石（211）面的晶格间距为0.27nm, 晶粒尺寸约为40nm.     

Evaluation of in vitro bioactivity and biocompatibility of Bioglass®-reinforced polyethylene composite

The bioactivity and biocompatibility of Bioglass®-reinforced high-density polyethylene composite (Bioglass®/HDPE) have been evaluated in simulated body fluid (SBF) and by in vitro cell culture, respectively. The formation of a biologically active hydroxy-carbonate apatite (HCA) layer on the composite surface after immersion in SBF was demonstrated by thin-film X-ray diffraction, infrared spectroscopy and scanning electron microscopy, indicating the in vitro bioactivity of Bioglass®/HDPE composites. The HCA layer was formed on the 40 vol% composite surface within 3 days immersion in SBF at a formation rate comparable to those on bioactive glass-ceramics, showing that in vitro bioactivity could be obtained in a composite. Furthermore, the composite was biocompatible to primary human osteoblast-like cells. In comparison with unfilled HDPE and tissue culture plastic control, a significant increase in cellular metabolic activity was found on the composite. Therefore, Bioglass®/HDPE composites have a promising biological response as a potential implant material.     

Preparation and bioactivity evaluation of hydroxyapatite-titania/chitosan-gelatin polymeric biocomposites

Biocomposites consisting of hydroxyapatite (HA) and natural polymers such as collagen, chitosan, chitin,and gelatin have been extensively investigated. However, studies on the combination of HA and titania with chitosan and gelatin have not been conducted yet. Novel biodegradable hydroxyapatite-titania/chitosan-gelatin polymeric composites were fabricated. In this work, our results are concerning with the preparation and characterization of HA powder and HA filler containing titania powder (10 and 30%) with a chitosan and gelatin copolymer matrix. The present research focuses on characterizing the structure of this novel class of biocomposites. Thermogravimetric analysis (TGA), X-ray diffraction (XRD), and Fourier Transformed Infrared Spectroscopy (FT-IR), Scanning electron microscopy (SEM-EDAX) were employed to assess the produced composites. The mechanical properties in terms of compressive strength and hardness test were also investigated. The in vitro study in simulated body fluid (SBF) was performed to assess the bioactivity of composites. The results proved that apatite resembling natural bone are formed faster and greater in the case the composite of HA containing 10% titania into chitosan-gelatin polymeric matrix when they are soaked in a simulated body fluid (SBF) than the composite containing 30% titania. The biocomposites containing HA with 10% titania are expected to be attractive for bioapplications as bone substitutes and scaffolds for tissue engineering in future.     

Synthesis and characterization of hydroxyapatite/chitosan nanocomposite materials for medical engineering applications

Incorporation of hydroxyapatite (HA) with organic polymer in favor of composites would be used in biomaterial engineering. According to prior researches, because of its chemical similarity to natural bone and dental, this product could improve bioactivity and bone bonding ability. In this research, nano-hydroxyapatite/chitosan composite material was prepared via in situ Hybridization route. The surface chemical characterization on the nanocomposite was evaluated by Fourier transformed infrared (FTIR) and X-ray diffraction (XRD). Surface topography, roughness and morphology of the samples were observed by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The characterization results confirmed homogeneity, interaction and integration between the HA and chitosan matrix. It was indicated that composite samples consist of homogeneous aggregations around 40–100 nm, in which many HA nanocrystals align along the chitosan molecules. HA grain gradually decreased in size when amount of chitosan increased from 0 to 6 g into 100 cc solution. It can be seen that by increasing chitosan, the aggregation of nanoparticles enhance and subsequently, improve the expected compatibility among HA filler and chitosan matrix. Furthermore, the mechanical compressive testing indicated that the synthesized composites have acceptable mechanical behavior for tissue substitution. The mechanistic of the biodegradable nanocomposite systems, their preparation and characterization for medical usage are strongly discussed.     

Hydroxyapatite/Bioactive Glass Films Produced by a Sol–Gel Method: In Vitro Behavior

Hydroxyapatite (HA) and HA/bioactive glass (49S) films were deposited on Si(100) substrates by a sol–gel dip‐coating method. The microstructure and in vitro bioactivity of the films were investigated by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and X‐ray photoelectron spectroscopy (XPS). Polycrystalline HA and amorphous bioactive glass films were obtained after annealing at 600 and 400 °C, respectively. The crystallization temperature of HA was determined to be around 568 °C. The surfaces of the HA films were covered with an apatite layer consists of spherulites formed by nanosized needle‐like aggregates after the soaking in simulated body fluid (SBF) for 10 days, while amorphous HA/bioactive glass surface was covered with larger spherical crystallites. Both XPS and EDS results obtained from HA/bioactive glass film, after soaking in SBF, showed increasing P amounts on the surface at the expense of Si. The higher density of the newly formed layer on HA/bioactive glass surface than that of the HA surface after 10 days of soaking was evidence of increased reaction rate and apatite forming ability when bioactive glass layer is present on the HA films.     

电气石/壳聚糖复合纤维的表征及细胞相容性

以壳聚糖为基体,电气石为分散相,采用溶液纺丝法制备电气石/壳聚糖复合纤维,利用光学显微镜、扫描电镜以及红外光谱仪对材料进行表征。电气石/壳聚糖复合纤维与人骨肉瘤细胞株（MG63）体外共培养,初步评价了材料的细胞相容性。结果显示,电气石颗粒在复合纤维中分散均匀且被壳聚糖包裹,纤维表面无裸露电气石。细胞在电气石/壳聚糖复合纤维表面黏附及生长增殖状况良好,材料对细胞无明显毒性。该材料有望成为一种良好的创伤修复敷料。     

Fabrication of fibrous poly(butylene succinate)/wollastonite/apatite composite scaffolds by electrospinning and biomimetic process

In this paper, a novel kind of Poly(butylene succinate) (PBSU) /wollastonite/apatite composite scaffold was fabricated via electrospinning and biomimetic process. Pure PBSU scaffold and composite scaffolds with 12.5 wt% and 25 wt% wollastonite were firstly fabricated by electrospinning. SEM micrographs showed that all the electrospun scaffolds had homogeneous fibrous structures with interconnected pores and randomly oriented ultrafine fibers. The composite scaffolds were then surface modified using a biomimetic process. SEM and XRD results showed that apatite could deposit on the surfaces of the composite fibers after incubation in SBF and a novel fibrous structure with microspheres composed of worm-like apatite on composite fibers was formed. Incubation time and wollastonite content were found to influence the morphology of the scaffolds during the biomimetic process obviously. Both the amount and the size of the microspheres on the composite scaffolds increased with increased incubation time. After a certain incubation time, microspheres formed on the composite fibers with less wollastonite had a relatively larger size. Therefore, the microstructure of the composite scaffolds could be adjusted by controlling the wollastonite content and the incubation time. All of these results suggest that it is an effective approach to fabricate PBSU/wollastonite/apatite fibrous composite scaffolds with different material content and controllable microstructure for bone tissue engineering.     

生物活性玻璃/PLA-PEG-PLA/聚乳酸组织工程支架在SBF溶液中的降解和矿化

通过测定pH值、质量损失率、SEM、XRD和FTIR,系统研究了生物活性玻璃/聚乳酸-聚乙二醇-聚乳酸嵌段共聚物（PLA-PEG-PLA）/聚乳酸组织工程支架在模拟体液（SBF）中的降解和生物矿化性能。研究结果表明：随着支架在SBF溶液中浸泡时间的延长,SBF的pH值和支架的质量呈下降趋势;生物活性玻璃的存在使pH值升高,而PLA-PEG-PLA嵌段共聚物的存在使pH值降低。XRD、FTIR图谱和SEM图像表明：在SBF中浸泡一定时间后,有无定型或结晶不完善的磷灰石在生物活性玻璃/PLA-PEG-PLA/聚乳酸组织工程支架表面沉积形成,并且PLA-PEG-PLA共聚物降解速度比聚乳酸快;在SBF中浸泡7天后,PLA-PEG-PLA共聚物的含量已经很难通过FTIR检测出来。