聚醚醚酮复合材料表面生物活性涂层的制备与性能

以聚醚醚酮/钡玻璃粉(PEEK-BGF)复合材料为基体, 通过硅烷偶联剂, 在复合材料表面构建具有生物活性的纳米羟基磷灰石(nHA)和甲基丙烯酸酯基的光固化树脂复合涂层. 采用扫描电子显微镜(SEM)和X射线光电子能谱(XPS)分析了材料表面形貌和元素分布, 测试了涂层与复合材料之间的粘接强度. 通过检测大鼠成骨细胞总蛋白含量和碱性磷酸酶表达水平, 评价新型光固化纳米羟基磷灰石/聚甲基丙烯酸酯(nHA/PMMA)复合涂层的生物活性. 研究结果表明, nHA填充的光固化复合材料形成粗糙的表面, 随着nHA的填充量提高, 涂层表面生物学活性得到提高.     

生物活性复合材料在生物矿化过程中的微观形貌与离子浓度变化

采用3-羟基丁酸酯-3-羟基戊酸酯的共聚物(PHBV)与磷酸三钙(TCP)、羟基磷灰石(HA)、生物活性玻璃(BG)等进行复合,获得了性能优良的生物活性骨组织工程支架材料和骨修复材料.研究和比较了3种复合材料在体外模拟生理环境中发生的生物矿化反应,对比了反应前后因矿化物的形成导致的表面微观形貌的变化,通过检测反应液离子浓度的变化对不同材料的矿化过程和生物活性进行了定量表征.研究结果表明,3种复合材料在模拟生理溶液中发生生物矿化的过程和程度不同,其中PHBV/BG的生物活性反应最为明显.     

纳米羟基磷灰石/丝素蛋白复合支架材料的降解特性及生物相容性研究

应用共混法制备了纳米羟基磷灰石/丝素蛋白复合支架材料, 通过体外降解和细胞培养实验研究了复合支架材料的降解特性和生物相容性. 体外降解实验结果显示, 复合支架材料具有稳定的降解能力; 在降解过程中, 羟基磷灰石由于与降解液发生钙、磷等离子的交换, 使其结晶得到了进一步生长和完善. 利用细胞计数法、四甲基偶氮唑盐(MTT)比色法和碱性磷酸酶(ALP)活性测定等分析了复合支架材料的生物相容性, 结果表明, MG63细胞在复合支架材料上具有良好的粘附、增殖能力, 并可引起早期的骨分化. 因此, 纳米羟基磷灰石/丝素蛋白复合支架作为骨组织工程的支架材料具有良好的应用前景.     

Incorporation of nanohydroxyapatite and vitamin D3 into electrospun PCL/Gelatin scaffolds: The influence on the physical and chemical properties and cell behavior for bone tissue engineering

Scaffold, an essential element of tissue engineering, should provide proper physical and chemical properties and evolve suitable cell behavior for tissue regeneration. Polycaprolactone/Gelatin (PCL/Gel)‐based nanocomposite scaffolds containing hydroxyapatite nanoparticles (nHA) and vitamin D3 (Vit D3) were fabricated using the electrospinning method. Structural and mechanical properties of the scaffold were determined by scanning electron microscopy (SEM) and tensile measurement. In this study, smooth and bead‐free morphology with a uniform fiber diameter and optimal porosity level with appropriate pore size was observed for PCL/Gel/nHA nanocomposite scaffold. The results indicated that adding nHA to PCL/Gel caused an increase of the mechanical properties of scaffold. In addition, chemical interactions between PCL, gelatin, and nHA molecules were shown with XRD and FT‐IR in the composite scaffolds. MG‐63 cell line has been cultured on the fabricated composite scaffolds; the results of viability and adhesion of cells on the scaffolds have been confirmed using MTT and SEM analysis methods. Here in this study, the culture of the osteoblast cells on the scaffolds showed that the addition of Vit D3 to PCL/Gel/nHA scaffold caused further attachment and proliferation of the cells. Moreover, DAPI staining results showed that the presence and viability of the cells were greater in PCL/Gel/nHA/Vit D3 scaffold than in PCL/Gel/nHA and PCL/Gel scaffolds. The results also approved increasing cell proliferation and alkaline phosphatase (ALP) activity for MG‐63 cells cultured on PCL/Gel/nHA/Vit D3 scaffold. The results indicated superior properties of hydroxyapatite nanoparticles and vitamin D3 incorporated in PCL/Gel scaffold for use in bone tissue engineering.     

Biomimetic polymer/apatite composite scaffolds for mineralized tissue engineering

The material surface must be considered in the design of scaffolds for bone tissue engineering so that it supports bone cells adhesion, proliferation and differentiation. A biomimetic approach has been developed as a 3D surface modification technique to grow partially carbonated hydroxyapatite (the bonelike mineral) in prefabricated, porous, polymer scaffolds using a simulated body fluid in our lab. For the rational design of scaffolding materials and optimization of the biomimetic process, this work focused on various materials and processing parameters in relation to apatite formation on 3D polymer scaffolds. The apatite nucleation and growth in the internal pores of poly(L-lactide) and poly(D,L-lactide) scaffolds were significantly faster than in those of poly(lactide-co-glycolide) scaffolds in simulated body fluids. The apatite distribution was significantly more uniform in the poly(L-lactide) scaffolds than in the poly(lactide-co-glycolide) scaffolds. After incubation in a simulated body fluid for 30 d, the mass of poly(L-lactide) scaffolds increased approximately 40%, whereas the mass of the poly(lactide-co-glycolide) scaffolds increased by about 15% (see Figure). A higher ionic concentration and higher pH value of the simulated body fluid enhanced apatite formation. The effects of surface functional groups on apatite nucleation and growth were found to be more complex in 3D scaffolds than on 2D films. Surprisingly enough, it was found that carboxyl groups significantly reduced the apatite formation, especially on the internal pore surfaces of 3D scaffolds. These findings are critically important in the rational selection of materials and surface design of 3D scaffolds for mineralized tissue engineering and may contribute to the understanding of biomineralization as well.SEM micrograph of a poly(L-lactide) scaffold.     

Superior corrosion protection and in vitro biocompatibility of Na-HAp/CS composite coating on PoPD-coated 316L SS

The synthesis and development of sodium (Na)-substituted hydroxyapatite (HAp)/chitosan (CS) composite using poly (O-phenylenediamine) (PoPD) coating on 316L SS substrate for improving bioactivity and corrosion protection was studied. The surface of Na-HAp/CS/PoPD bilayer coatings on 316L SS substrate was characterized by diverse analytical techniques. The open circuit potential (OCP) measurement, potentiodynamic polarization, and impedance test revealed that the bilayer coating provides excellent protection to the substrate against the corrosion in the simulated body fluid (SBF) solution. This interior layer of the coating acts as a barrier against the release of metal ions from the substrate, which was confirmed by inductively coupled plasma-atomic emission spectroscopy. Besides, the mechanical properties of the coatings were analyzed. From the obtained results, the bilayer coating exhibited greater mechanical strength than the individual coating. An in vitro bioactivity of the coatings was assessed by immersion in the SBF solution at 7–28 days. The apatite formation of bilayer coatings on 316L SS substrate is found to be more bioactive compared with the Na-HAp, PoPD, and Na-HAp/CS. The in vitro biocompatibility test showed no adverse effects, which was proved by the enhanced biocompatibility of the bilayer coating on 316L SS.     

Interfacial Compatibilization into PLA/Mg Composites for Improved In Vitro Bioactivity and Stem Cell Adhesion

The present work highlights the crucial role of the interfacial compatibilization on the design of polylactic acid (PLA)/Magnesium (Mg) composites for bone regeneration applications. In this regard, an amphiphilic poly(ethylene oxide-b-L,L-lactide) diblock copolymer with predefined composition was synthesised and used as a new interface to provide physical interactions between the metallic filler and the biopolymer matrix. This strategy allowed (i) overcoming the PLA/Mg interfacial adhesion weakness and (ii) modulating the composite hydrophilicity, bioactivity and biological behaviour. First, a full study of the influence of the copolymer incorporation on the morphological, wettability, thermal, thermo-mechanical and mechanical properties of PLA/Mg was investigated. Subsequently, the bioactivity was assessed during an in vitro degradation in simulated body fluid (SBF). Finally, biological studies with stem cells were carried out. The results showed an increase of the interfacial adhesion by the formation of a new interphase between the hydrophobic PLA matrix and the hydrophilic Mg filler. This interface stabilization was confirmed by a decrease in the damping factor (tanδ) following the copolymer addition. The latter also proves the beneficial effect of the composite hydrophilicity by selective surface localization of the hydrophilic PEO leading to a significant increase in the protein adsorption. Furthermore, hydroxyapatite was formed in bulk after 8 weeks of immersion in the SBF, suggesting that the bioactivity will be noticeably improved by the addition of the diblock copolymer. This ceramic could react as a natural bonding junction between the designed implant and the fractured bone during osteoregeneration. On the other hand, a slight decrease of the composite mechanical performances was noted.     

A comparative study of the in vitro degradation of poly(l-lactic acid)/β-tricalcium phosphate scaffold in static and dynamic simulated body fluid

Porous poly(l-lactic acid)/β-tricalcium phosphate (PLLA/β-TCP) composite is a new promising scaffold for bone tissue engineering. Porous scaffolds fabricated by liquid anti-solvent precipitation principle were subjected to degradation in dynamic simulated body fluid (DSBF) and in static simulated body fluid (SSBF) at 37 °C for 24 weeks, respectively. Results indicated that a large number of apatite layer were formed on the scaffolds. The results further indicated that SBF flow decreased the degradation rate of molecular weight and compressive strength significantly. The porosity and mass changes were related to the apatite formation and SBF flow. All the results might be owed to the mutual effects of the flow of SBF and the addition of β-TCP. The degradation rate of scaffolds could be adjusted by the additional fraction of β-TCP to meet the requirements of application in vivo.     

A study on biomineralization behavior of N-methylene phosphochitosan scaffolds

Biomimetic growth of calcium phosphate over natural polymer may be an effective approach to constituting an organic/inorganic composite scaffold for bone tissue engineering. In this work, N-methylene phosphochitosan (NMPCS) was prepared via formaldehyde addition and condensation with phosphoric acid in a step that allowed homogeneous modification without obvious deterioration in chitosan (CS) properties. The NMPCS obtained was characterized by using FT-IR and elemental analysis. The macroporous scaffolds were fabricated through a freeze-drying technique. A comparative study on NMPCS and CS scaffold biomimetic mineralization was carried out in different media, i.e, a simulated body fluid (SBF) or alternative CaCl(2) and Na(2)HPO(4) solutions respectively. Apatite formation within NMPCS and CS scaffolds was identified with FT-IR, scanning electron microscopy (SEM), energy-dispersive X-ray (EDX) and X-ray diffractometery (XRD). The results revealed alternate soaking of the scaffolds in CaCl(2) and Na(2)HPO(4) solutions was better than soaking in SBF solution alone in relation to apatite deposition on the scaffold pore walls. Biomineralization provides an approach to improve nature derived materials, e.g., chitosan derivative NMPCS properties e.g., compressive modulus, etc. SEM image of a NMPCS/apatite composite scaffold.     

羟基磷灰石/氧化铁复合涂层的制备及其诱导骨生长的生物活性

用电泳沉积法制得羟基磷灰石/壳聚糖/氧化铁(HA/CS/Fe2O3)复合涂层,经700 ℃烧结处理得到HA/Fe2O3复合涂层。 通过SEM、EDS、XRD、FT-IR、电化学和万能材料试验机等对复合涂层的表面形貌、物相组成、抗腐蚀性和结合强度进行了表征和测试,最后采用1.5SBF浸泡法对复合涂层的生物活性进行了评价。 结果表明,当悬浮液中的HA、CS与Fe2O3质量比为100∶100∶1时,所制得的HA/Fe2O3复合涂层表面粗糙,抗腐蚀性强,具有良好的诱导骨生长生物活性,基体与复合涂层结合强度可达27.5 MPa。     

Effects of Laminin For Osteogenesis in Porous Hydroxyapatite

Summery: As a tooth is composed of hard tissue covering pulp, it may be suitable for tooth regeneration to use porous cylindrical hydroxyapatite (HA) scaffolds with a hollow center. Generally, in vivo examination, bone marrow cell suspension for osteogenesis in cell/HA composite scaffold without subculture is prepared at a density of 1 × 10⁷ cells/ml or higher. In dentistry, stem cells would be obtained from tooth pulp. For dentine formation, a smaller number of stem cells must be used. In this study, a suspension of rat bone marrow cells at 1 × 10⁶ cells/ml of density was prepared to estimate the adhesive effect of laminin. After immersion of HA scaffold in laminin solution, bone marrow cells were seeded in the pores of the HA scaffolds by immersion in the cell suspension for preparing the cell/HA composite scaffolds. The specimens were respectively implanted in the dorsal subcutis of 7-week-old male Fischer 344 rats for 4 weeks for histological examination. Comparing with the results of in vivo examination, alkaline phosphatase activity of bone marrow cells on laminin-coated plate with and without dexamethasone cultured for 2 weeks was measured in vitro. It was considered that laminin contributed to bone formation in pores of a scaffold.     

Homogeneous organic/inorganic hybrid scaffolds with high osteoinductive activity for bone tissue engineering

Composite scaffolds of polymers/β-tricalcium phosphate (TCP) have been widely used for bone regeneration due to the combination of osteoinductivity of TCP and mechanical properties of the polymers. However, the difference in surface properties of the two material causes composite has poor uniformity and weak two-phase interaction, resulting in poor TCP release and weak new bone-forming ability. In this research, a TCP sol was developed to replace traditional TCP nanoparticles for the preparation of homogeneous polycaprolactone (PCL)/TCP sol nanofibrous scaffolds. It was found that compared with TCP nanoparticles, TCP sol homogeneously distributed in PCL nanofibers, and greatly improved the hydrophilicity, biodegradability, and mechanical properties of the scaffolds. It is also confirmed that loading TCP sol promoted the formation of bone-like apatite on the surface of the scaffolds. Biological experiments showed that all scaffolds supported rat bone marrow mesenchymal stem cells (rBMSCs) proliferation, especially scaffolds loaded with TCP sol. The increase in alkaline phosphatase activity and collagen production, enhanced calcium deposition, and up-regulation of osteocalcin expression demonstrated that the loading TCP sol expanded an advantage of scaffolds in promoting rBMSCs osteogenic differentiation, suggesting it dramatically improved the osteoinductive activity of PCL/TCP hybrid system and had a great potential application in bone regeneration.     

钛基多孔HA/SiO2复合涂层的制备及性能研究

为了制得表面多孔且与基材结合强度高的羟基磷灰石(HA)涂层,实验中以正丁醇为分散介质,以SiO2粉末为添加剂,纯钛片为基材,电泳沉积制备羟基磷灰石/二氧化硅/壳聚糖/(HA/SiO2/CS)复合涂层,经后续热处理得到多孔HA/SiO2复合涂层,采用扫描电镜(SEM)、傅立叶红外光谱仪(FT-IR)、X射线衍射仪(XRD)、万能材料试验机对涂层的表面形貌、组成、结构和结合强度进行测试和表征,并通过模拟体液(SBF)浸泡法对复合涂层的生物活性进行评价.结果表明:当悬浮液中的HA/SiO2/CS质量比为1∶1∶1时,制得的HA/SiO2/CS涂层经700℃热处理后获得的HA/SiO2复合涂层孔洞分布均匀,大孔孔径在10～15μm,小孔孔径在1～5μm;涂层与基材的结合强度达到25.5 MPa;多孔HA/SiO2复合涂层在SBF中浸泡7 d后,涂层表面碳磷灰石化;说明实验中添加SiO2所制得的多孔HA/SiO2复合涂层与钛基材结合强度高,且具有良好的生物活性.     

Bioactivation of knitted cellulose scaffolds by strontium

Strontium has attained increasing interest in the treatment of osteoporosis due to its anabolic as well as antiresorptive activity. Knitted cellulose scaffolds with a porosity suitable to promote bone tissue ingrowth and vascularisation were doped with SrCO₃. The in vitro bioactivity of the modified scaffolds was proven by formation of hydroxyapatite during exposure to simulated body fluid. The Sr-release kinetics during static exposure to simulated body fluid is dominated by an accelerated Sr-release in the initial state followed by a reduced release corresponding to a diffusion controlled rate. Microstructural analyses indicate that initially precipitated SrCO₃ transforms to Sr _x Ca_1−x CO₃ solid solutions that subsequently serve as a template for the precipitation of bone like carbonated hydroxyapatite under conditions simulating the inorganic part of the human blood plasma.     

Interaction of acid and alkali treated titanium with dynamic simulated body environment

Interaction of acid and acid+alkali treated titanium samples with simulated body fluid was studied. In case of alkali treated titanium, the dynamic arrangement of the test enabled the detection of primary calcium and phosphate ion adsorption from the solution and later apatite crystal growth (XRD). The induction time for crystal growth was 24.2±0.3 h. On acid-only treated titanium no crystal growth was detected. The calcium phosphate adsorption layer formed on the acid treated samples was detectable by XPS only, however it differed from that one formed on the acid+alkali treated samples. The adsorption layer formed on the acid+alkali treated samples contained larger amount of calcium, especially in the shortest exposure times. Charging of the apatite crystallites during the XPS measurement enabled the determination their Ca/P ratio separately from Ca/P ratio of the adsorption layers. XPS and EDS analyses indicated that the spherulitic crystallites consisted of carbonated hydroxyapatite with the Ca/P ratio close to that one of the stoichiometric hydroxyapatite. It is proposed that the adsorption layer formed spontaneously and immediately on the acid+alkali treated titanium can provide an ideal interface between the metal implant and the apatite cement line, the first structure formed by osteoblast cells during the formation of the new bone on foreign surfaces. This revised version was published online in August 2006 with corrections to the Cover Date.     

Fabrication of poly(lactide‐co‐glycolide) scaffold embedded spatially with hydroxyapatite particles on pore walls for bone tissue engineering

Poly(lactide‐co‐glycolide) (PLGA) scaffolds embedded spatially with hydroxyapatite (HA) particles on the pore walls (PLGA/HA‐S) were fabricated by using HA‐coated paraffin spheres as porogens, which were prepared by Pickering emulsion. For comparisons, PLGA scaffolds loaded with same amount of HA particles (2%) in the matrix (PLGA/HA‐M) and pure PLGA scaffolds were prepared by using pure paraffin spheres as porogens. Although the three types of scaffolds had same pore size (450–600 µm) and similar porosity (90%–93%), the PLGA/HA‐S showed the highest compression modulus. The embedment of the HA particles on the pore walls endow the PLGA/HA‐S scaffold with a stronger ability of protein adsorption and mineralization as well as a larger mechanical strength against compression. In vitro culture of rat bone marrow stem cells revealed that cell morphology and proliferation ability were similar on all the scaffolds. However, the alkaline phosphatase activity was significantly improved for the cells cultured on the PLGA/HA‐S scaffolds. Therefore, the method for fabricating scaffolds with spatially embedded nanoparticles provides a new way to obtain the bioactive scaffolds for tissue engineering. Copyright © 2011 John Wiley & Sons, Ltd.     

碳纳米管/壳聚糖复合微球的原位仿生矿化及表征

对碳纳米管(CNTs)进行酸化处理, 采用乳化交联法制备CNTs/壳聚糖(CS)复合微球, 在其表面诱导羟基磷灰石仿生合成, 研究了CNTs对复合微球仿生矿化的影响, 并与纯CS微球的仿生矿化进行了对比. 利用扫描电子显微镜(SEM)、 X射线衍射仪(XRD)、 溶胀率和含水率测试等考察了复合微球矿化前后的形貌特征、 物相结构及稳定性. 结果表明, 在相同时间下, CNTs/CS复合微球表面纳米羟基磷灰石的形成能力明显优于纯CS微球, 且形态稳定性更高. 细胞实验结果表明, 与MG63细胞共培养7 d时, 矿化复合微球细胞增殖明显.     

Production and characterization of chitosan fibers and 3-D fiber mesh scaffolds for tissue engineering applications

This study reports on the production of chitosan fibers and 3-D fiber meshes for the use as tissue engineering scaffolds. Both structures were produced by means of a wet spinning technique. Maximum strain at break and tensile strength of the developed fibers were found to be 8.5% and 204.9 MPa, respectively. After 14 d of immersion in simulated body fluid (SBF), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), and inductively coupled plasma emission (ICP) spectroscopy analyses showed that a bioactive Ca-P layer was formed on the surface of the fibers, meaning that they exhibit a bioactive behavior. The samples showed around 120% max. swelling in physiological conditions. The pore sizes of 3-D chitosan fiber mesh scaffolds were observed to be in the range of 100-500 microm by SEM. The equilibrium-swelling ratio of the developed scaffolds was found to be around 170% (w/w) in NaCl solution at 37 degrees C. Besides that, the limit swelling strain was less than 30%, as obtained by mechanical spectroscopy measurements in the same conditions. The viscoelastic properties of the scaffolds were also evaluated by both creep and dynamic mechanical tests. By means of using short-term MEM extraction test, both types of structures (fibers and scaffolds) were found to be non-cytotoxic to fibroblasts. Furthermore, osteoblasts directly cultured over chitosan fiber mesh scaffolds presented good morphology and no inhibition of cell proliferation could be observed.Osteoblast-like cells proliferating over chitosan based fibers after 7 d of culture.     

Fabrication and properties of mineralized collagen‐chitosan/hydroxyapatite scaffolds

A hydroxyapatite (HAp)/biopolymer composite scaffold was fabricated by mineralizing a crosslinked collagen/chitosan, which was pre‐mineralized with Ca²⁺ and phosphate salts, in simulated body fluid (SBF) for only 24 hr. A self‐organized structure similar to bone is expected. Microstructures of the crosslinked collagen/chitosan scaffold, the pre‐mineralized collagen–chitosan scaffold (CCS), and the mineralized collagen‐chitosan/HAp scaffolds (MCCHS) were characterized by scanning electron microscopy (SEM), revealing non‐alteration of the porous structure and formation of the HAp particles. X‐ray diffractometer (XRD) confirmed the crystalline structure of the HAp. Thermal gravimetric analysis found that more HAp particles were formed when the CCSs were pre‐mineralized in a higher concentration of Ca²⁺. Water‐uptake ratio of the crosslinked CCS was ～160, decreased to ～120 after incubating in Ca²⁺ solution, and further decreased to ～20 after mineralization. Mechanical strength of the CCS was improved significantly after the in situ mineralization too. The method introduced here may be potentially applied to obtain other biopolymer/HAp composite in a short period. Copyright © 2008 John Wiley & Sons, Ltd.     

In Vitro Biomineralization and Bulk Characterization of Chitosan/Hydroxyapatite Composite Microparticles Prepared by Emulsification Cross-Linking Method: Orthopedic Use

Chitosan/hydroxyapatite composite microparticles were prepared by a solid-in-water-in-oil emulsification cross-linking method. The characteristics and activity in presence of simulated body fluid for 14 and 21?days were investigated. The size distribution, surface morphology, and microstructure of these biomaterials were evaluated. The scanning electron microscopy revealed an aggregate of microparticles with a particle size, ranged from 4 to 10???m. The deposited calcium phosphate was studied using X-ray diffraction analysis, Fourier transform infrared spectroscopy, and inductively coupled plasma/atomic emission?spectroscopy analysis of phosphorus. These results show that the mineral, formed on microparticles, was a mixture of carbonated hydroxyapatite and calcite. Scanning electron microscopy revealed that calcium phosphate crystals growth was in form of rods organized as concentric triangular packets interconnected to each other by junctions. Interaction between chitosan and growing carbonated hydroxyapatite and calcite crystals are responsible for a composite growth into triangular and spherical shapes. The results demonstrated that these microparticles were potential materials for bone repair.