钛基羟基磷灰石复合涂层的研究
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摘要
羟基磷灰石(Hydroxyapatite, HA)涂层由于具有良好的生物相容性和骨诱导性,被广泛的研究和应用于临床。近年来,将HA与其它生物材料复合,从而改善HA涂层的性能,满足不同的临床需要,成为生物材料领域的研究热点。目前制备的HA复合涂层主要分为HA-有机复合涂层和HA-无机复合涂层。HA-有机复合涂层主要是将HA与生物大分子复合以期获得优良生物学性能,例如壳聚糖(Chitosan, CS),蛋白等。HA-无机复合涂层主要是将HA与无机材料,如ZrO2, TiO2等复合,以改善涂层的力学性能。然而复合涂层不均匀性一直是困扰其走向应用的主要问题。本研究制备了HA与壳聚糖(CS)、明胶(Gelatin)、氧化锆(ZrO2)复合涂层。通过比较脉冲电位和恒电位,不同脉冲电位,电解液成分对涂层结晶、成分、形貌等影响来优化工艺,然后对复合涂层进行了生物学和力学性能评价。采用扫描电子显微镜(SEM)、X射线衍射仪(XRD)、傅立叶红外光谱仪(FTIR)、热重分析仪(TG)和X射线光电子能谱仪(XPS)等分析手段,研究了HA复合涂层的结构组成、形貌和性能。主要内容如下：
采用电化学沉积法在钛金属表面制备出羟基磷灰石／壳聚糖(HA/CS)复合涂层,实现CS与HA在微观尺寸上的复合与杂化。比较了脉冲电位与恒电位模式下复合涂层的形成,研究了电位高低及壳聚糖浓度对复合涂层性能的影响。结果表明,与恒电位模式比较,脉冲电位下制备的涂层较均匀、结晶性好、CS含量高,并且HA与CS杂化程度高。脉冲电位的高低影响涂层的结晶、形貌；脉冲低电位(-1.3 V)更利于复合涂层的沉积。电解液中CS浓度影响复合涂层的形貌及CS在复合涂层中的含量,电解液中CS浓度为0.33 g／l左右比较适合。抗菌检测表明复合涂层具有良好的抗菌性。成骨细胞的培养结果表明复合涂层具有良好的生物相容性。
通过原位水热合成和溶胶-凝胶浸提涂覆法在碱处理的钛表面制备了HA/CS复合涂层。接触角检测表明碱处理使钛表面具有超亲水性,这是形成均匀涂层的必要条件。X射线衍射分析表明复合涂层成分为HA和CS,各组分含量由热重分析确定。扫描电镜对复合涂层的形貌进行观察并发现不同HA含量的复合涂层具有不同的形貌。通过培养成骨细胞考察了复合涂层的细胞相容性。Alamar Blue检测表明HA/CS复合涂层表面细胞黏附及增殖能力较好。碱性磷酸酶(ALP)检测表明HA／CS复合涂层表面的细胞分化能力较好。综合研究结果表明该复合涂层有较好的生物相容性。
采用电化学沉积法制备磷酸钙／明胶(CaP/Gelatin)复合涂层,对其进行表征及性能测试。讨论了不同脉冲电位、明胶浓度、沉积温度、电解液pH值等对复合涂层性能的影响。研究发现脉冲电位下涂层质量优于恒电位。XRD和FTIR分析表明复合涂层的成分为HA和明胶。SEM观察发现复合涂层是结构均匀的多孔网状或花瓣状微纳结构。复合涂层形貌及成分随脉冲电位、明胶浓度等实验条件的变换而变化。低电位利于复合涂层的结晶和沉积。改变明胶在钙磷电解液中的浓度,发现明胶浓度在0.1～1.0g/l范围内不影响涂层形貌,但影响明胶在复合涂层中的含量。沉积温度为50℃,电解液pH值在明胶等电点以下保证了复合涂层具有较好的结晶性、均匀性。Alamar Blue检测表明复合涂层具有较好的生物相容性。
采用电化学沉积法在生物医用钛金属表面制备出均匀的HA/ZrO2复合涂层。通过热处理提高涂层的致密性,同时保留涂层的微纳结构。考察了热处理后复合涂层的成分、形貌、生物相容性及稳定性。X射线衍射分析表明复合涂层成分为HA和ZrO2。扫描电镜对HA/ZrO2复合涂层的形貌进行观察并发现热处理后复合涂层的致密性提高。研究发现ZrO2的加入大大降低了HA/ZrO2复合涂层中钙离子的释放速度,提高了HA/ZrO2复合涂层的稳定性。模拟体液(SBF)浸泡检测表明HA/ZrO2复合涂层能够诱导磷灰石生成,具有较好的生物活性。纳米划痕实验和拉力测试结果表明HA/ZrO2复合涂层具有较好的结合强度。通过培养成骨细胞考察了复合涂层的生物相容性。Alamar Blue检测表明HA/ZrO2复合涂层表面细胞黏附及增殖能力较好。ALP检测表明热处理后HA/ZrO2复合涂层表面的细胞分化能力较强。综合细胞培养结果表明HA/ZrO2复合涂层有较好的生物相容性。
综上所述,本研究成功制备出HA/CS, HA/Gelatin, HA/ZrO2复合涂层,为进一步研究HA复合涂层在骨缺损修复中的应用提供依据。
Bioactive hydroxylapatite (HA) coatings are commonly used to improve the biocompatibility and osteoinductivity of the titanium. HA coated titanium has the advantages of metals and ceramics and has been considered as one of the most promising bone replacement materials. Recently, there is a tendency to develop HA-contained composite coatings to obtain new type of coatings that satisfy the different requirement of clinic applications. One type of the composite coatings is to incorporate biomacromolecules, such as chitosan (CS) and gelatin, into HA coatings in order to promote the biological performance of the coatings. The other is to add inorganic materials, such as ZrO2 and TiO2, into HA coatings in order to improve the mechanical properties of coatings. The homogeneity of composite coatings is still a big challenge. In this study, three types of composite coatings, HA/CS, HA/geltin and HA/ZrO2, were prepared and the processing parameters was optimized. The microstructure, composition and biological performance of coating was evaluated. The main findings were summarized as follows.
Chitosan (CS) is a biopolymer that has good biocompatibility, antibacterial properties and the suitability for cell growth. In the present study, the electrochemical deposition method was-employed to produce HA/CS composite coatings on titanium substrates. The results indicated that pulse voltage mode can improve the CS content in the coatings and can obtain uniform coatings, compared with constant voltage mode. Pulse voltage mode also promoted the interaction between Ca and CS according to XPS analysis. Pulse voltage affected the morphology of the coatings and efficiency of the deposition. The suitable pulse voltage is-1.3 V. The concentration of CS in the electrolyte affected the CS content in the coatings and the efficiency of the deposition. The suitable one is 33.1g/l. The antibacterial test indicated that HA/CS coatings had good bactericidal ability. Osteoblasts were cultured on the coatings to evaluate the biocompatibility of coatings and the results indicated that the composite coatings could favor the attachment and proliferation of osteoblasts.
HA/CS composite coatings were also prepared on titanium surfaces by hydro-thermal synthesis and sol-gel method. The composition, structure, morphology and biocompatibility of composite coatings were characterized. Alkali treatment converted Ti surfaces into super-hydrophilic surfaces, which was the essential prerequisite for preparing uniform HA/CS composite coatings. XRD analysis revealed that the coatings were mainly composed of HA and CS and the content of composition were determined by TG analysis. SEM observation found the coatings with different HA content had different morphologies. Osteoblasts were cultured on the coatings to evaluate their biocompatibility. Alamar Blue assay indicated that cells on the composite coatings had higher proliferation rate than those on pure titanium. Also the ALP activity of the cells on the composite coatings was higher than those on the pure titanium samples. These results demonstrated that as-prepared HA/CS composite coatings have good biocompatibility.
Gelatin is the denatured form of collagen and is expected to be beneficial for hard tissue applications. The CaP/gelatin composite coatings could combine the bioactivity and osteoconductivity of CaP with the good characteristics of gelatin. In the present study, the pulsed electrochemical deposition method was employed to deposit CaP/gelatin composite coatings on titanium substrates. The main purpose of the study is to optimize the processing parameters of the composite coating. Electrolyte was the mixture of the Ca-P and gelatin aqueous solution with different concentration. SEM, XRD and FTIR were used to characterize the composition and morphology of the coating. The coatings prepared under constant voltage and pulse voltage electrochemical deposition were compared. It was found that the pulse voltage mode was more suitable for preparing CaP/gelatin coatings. The results indicated that the optimized experimental conditions were:pulse voltage range, 0～-1.3 V; the concentration of Ca2+,5.0×10-4mol/l; gelatin solution,0.5 g/l; pH value,5.0; temperature,50℃. Osteoblasts were cultured on the coatings to evaluate the biocompatibility of the coatings. Alamar Blue assay indicated that the composite coatings could favor the proliferation of the osteoblasts.
In recent years, HA/ZrO2 composite coatings have attracted extensive research attention because they could combine unique biocompatibility and bioactivity of HA ceramics with excellent mechanical properties of ZrO2. In this study, dense and uniform HA/ZrO2 composite coatings was fabricated on titanium substrates by pulsed electrochemical deposition. The composition, morphologies, biocompatibility and physiological stability of the composite coatings were studied. X-ray diffraction showed that OCP in the coating was converted into HA and the coating was composed of HA and ZrO2 after heat treatments. Scanning electron microscopy indicated that ZrO2 was uniformly distributed in the coatings and the coatings became dense after heat treatments. Simulated body fluid immersion test proved that the as-prepared composite coatings had good bioactivity to induce calcium phosphate formation under simulated physiological environment. Atomic absorption spectrometry analysis was used to measure Ca ion release rate of the coatings immersed in PBS. It was found that the release rate of Ca ion of HA/ZrO2 was lower than that of pure HA coatings. Nanoscratching tests and tensile tests revealed that HA/ZrO2 coatings had better interfacial bonding strength than that of pure HA coatings. Osteoblasts were cultured on the coatings to evaluate the biocompatibility of coatings. The results of the Alamar blue and ALP test indicated that the composite coatings could favor the proliferation and differentiation of the osteoblasts, which indicated that the as-prepared HA/ZrO2 nanocomposite coatings had good biocompatibility.
In summary, three types of HA-contained composite coatings, HA/CS, HA/geltin and HA/ ZrO2, have been successfully produced on titanium substrates. The results of present study provided valuable reference for the further research in developing composite coatings for biomedical applications.
采用电化学沉积法在钛金属表面制备出羟基磷灰石／壳聚糖(HA/CS)复合涂层,实现CS与HA在微观尺寸上的复合与杂化。比较了脉冲电位与恒电位模式下复合涂层的形成,研究了电位高低及壳聚糖浓度对复合涂层性能的影响。结果表明,与恒电位模式比较,脉冲电位下制备的涂层较均匀、结晶性好、CS含量高,并且HA与CS杂化程度高。脉冲电位的高低影响涂层的结晶、形貌；脉冲低电位(-1.3 V)更利于复合涂层的沉积。电解液中CS浓度影响复合涂层的形貌及CS在复合涂层中的含量,电解液中CS浓度为0.33 g／l左右比较适合。抗菌检测表明复合涂层具有良好的抗菌性。成骨细胞的培养结果表明复合涂层具有良好的生物相容性。
通过原位水热合成和溶胶-凝胶浸提涂覆法在碱处理的钛表面制备了HA/CS复合涂层。接触角检测表明碱处理使钛表面具有超亲水性,这是形成均匀涂层的必要条件。X射线衍射分析表明复合涂层成分为HA和CS,各组分含量由热重分析确定。扫描电镜对复合涂层的形貌进行观察并发现不同HA含量的复合涂层具有不同的形貌。通过培养成骨细胞考察了复合涂层的细胞相容性。Alamar Blue检测表明HA/CS复合涂层表面细胞黏附及增殖能力较好。碱性磷酸酶(ALP)检测表明HA／CS复合涂层表面的细胞分化能力较好。综合研究结果表明该复合涂层有较好的生物相容性。
采用电化学沉积法制备磷酸钙／明胶(CaP/Gelatin)复合涂层,对其进行表征及性能测试。讨论了不同脉冲电位、明胶浓度、沉积温度、电解液pH值等对复合涂层性能的影响。研究发现脉冲电位下涂层质量优于恒电位。XRD和FTIR分析表明复合涂层的成分为HA和明胶。SEM观察发现复合涂层是结构均匀的多孔网状或花瓣状微纳结构。复合涂层形貌及成分随脉冲电位、明胶浓度等实验条件的变换而变化。低电位利于复合涂层的结晶和沉积。改变明胶在钙磷电解液中的浓度,发现明胶浓度在0.1～1.0g/l范围内不影响涂层形貌,但影响明胶在复合涂层中的含量。沉积温度为50℃,电解液pH值在明胶等电点以下保证了复合涂层具有较好的结晶性、均匀性。Alamar Blue检测表明复合涂层具有较好的生物相容性。
采用电化学沉积法在生物医用钛金属表面制备出均匀的HA/ZrO2复合涂层。通过热处理提高涂层的致密性,同时保留涂层的微纳结构。考察了热处理后复合涂层的成分、形貌、生物相容性及稳定性。X射线衍射分析表明复合涂层成分为HA和ZrO2。扫描电镜对HA/ZrO2复合涂层的形貌进行观察并发现热处理后复合涂层的致密性提高。研究发现ZrO2的加入大大降低了HA/ZrO2复合涂层中钙离子的释放速度,提高了HA/ZrO2复合涂层的稳定性。模拟体液(SBF)浸泡检测表明HA/ZrO2复合涂层能够诱导磷灰石生成,具有较好的生物活性。纳米划痕实验和拉力测试结果表明HA/ZrO2复合涂层具有较好的结合强度。通过培养成骨细胞考察了复合涂层的生物相容性。Alamar Blue检测表明HA/ZrO2复合涂层表面细胞黏附及增殖能力较好。ALP检测表明热处理后HA/ZrO2复合涂层表面的细胞分化能力较强。综合细胞培养结果表明HA/ZrO2复合涂层有较好的生物相容性。
综上所述,本研究成功制备出HA/CS, HA/Gelatin, HA/ZrO2复合涂层,为进一步研究HA复合涂层在骨缺损修复中的应用提供依据。
Bioactive hydroxylapatite (HA) coatings are commonly used to improve the biocompatibility and osteoinductivity of the titanium. HA coated titanium has the advantages of metals and ceramics and has been considered as one of the most promising bone replacement materials. Recently, there is a tendency to develop HA-contained composite coatings to obtain new type of coatings that satisfy the different requirement of clinic applications. One type of the composite coatings is to incorporate biomacromolecules, such as chitosan (CS) and gelatin, into HA coatings in order to promote the biological performance of the coatings. The other is to add inorganic materials, such as ZrO2 and TiO2, into HA coatings in order to improve the mechanical properties of coatings. The homogeneity of composite coatings is still a big challenge. In this study, three types of composite coatings, HA/CS, HA/geltin and HA/ZrO2, were prepared and the processing parameters was optimized. The microstructure, composition and biological performance of coating was evaluated. The main findings were summarized as follows.
Chitosan (CS) is a biopolymer that has good biocompatibility, antibacterial properties and the suitability for cell growth. In the present study, the electrochemical deposition method was-employed to produce HA/CS composite coatings on titanium substrates. The results indicated that pulse voltage mode can improve the CS content in the coatings and can obtain uniform coatings, compared with constant voltage mode. Pulse voltage mode also promoted the interaction between Ca and CS according to XPS analysis. Pulse voltage affected the morphology of the coatings and efficiency of the deposition. The suitable pulse voltage is-1.3 V. The concentration of CS in the electrolyte affected the CS content in the coatings and the efficiency of the deposition. The suitable one is 33.1g/l. The antibacterial test indicated that HA/CS coatings had good bactericidal ability. Osteoblasts were cultured on the coatings to evaluate the biocompatibility of coatings and the results indicated that the composite coatings could favor the attachment and proliferation of osteoblasts.
HA/CS composite coatings were also prepared on titanium surfaces by hydro-thermal synthesis and sol-gel method. The composition, structure, morphology and biocompatibility of composite coatings were characterized. Alkali treatment converted Ti surfaces into super-hydrophilic surfaces, which was the essential prerequisite for preparing uniform HA/CS composite coatings. XRD analysis revealed that the coatings were mainly composed of HA and CS and the content of composition were determined by TG analysis. SEM observation found the coatings with different HA content had different morphologies. Osteoblasts were cultured on the coatings to evaluate their biocompatibility. Alamar Blue assay indicated that cells on the composite coatings had higher proliferation rate than those on pure titanium. Also the ALP activity of the cells on the composite coatings was higher than those on the pure titanium samples. These results demonstrated that as-prepared HA/CS composite coatings have good biocompatibility.
Gelatin is the denatured form of collagen and is expected to be beneficial for hard tissue applications. The CaP/gelatin composite coatings could combine the bioactivity and osteoconductivity of CaP with the good characteristics of gelatin. In the present study, the pulsed electrochemical deposition method was employed to deposit CaP/gelatin composite coatings on titanium substrates. The main purpose of the study is to optimize the processing parameters of the composite coating. Electrolyte was the mixture of the Ca-P and gelatin aqueous solution with different concentration. SEM, XRD and FTIR were used to characterize the composition and morphology of the coating. The coatings prepared under constant voltage and pulse voltage electrochemical deposition were compared. It was found that the pulse voltage mode was more suitable for preparing CaP/gelatin coatings. The results indicated that the optimized experimental conditions were:pulse voltage range, 0～-1.3 V; the concentration of Ca2+,5.0×10-4mol/l; gelatin solution,0.5 g/l; pH value,5.0; temperature,50℃. Osteoblasts were cultured on the coatings to evaluate the biocompatibility of the coatings. Alamar Blue assay indicated that the composite coatings could favor the proliferation of the osteoblasts.
In recent years, HA/ZrO2 composite coatings have attracted extensive research attention because they could combine unique biocompatibility and bioactivity of HA ceramics with excellent mechanical properties of ZrO2. In this study, dense and uniform HA/ZrO2 composite coatings was fabricated on titanium substrates by pulsed electrochemical deposition. The composition, morphologies, biocompatibility and physiological stability of the composite coatings were studied. X-ray diffraction showed that OCP in the coating was converted into HA and the coating was composed of HA and ZrO2 after heat treatments. Scanning electron microscopy indicated that ZrO2 was uniformly distributed in the coatings and the coatings became dense after heat treatments. Simulated body fluid immersion test proved that the as-prepared composite coatings had good bioactivity to induce calcium phosphate formation under simulated physiological environment. Atomic absorption spectrometry analysis was used to measure Ca ion release rate of the coatings immersed in PBS. It was found that the release rate of Ca ion of HA/ZrO2 was lower than that of pure HA coatings. Nanoscratching tests and tensile tests revealed that HA/ZrO2 coatings had better interfacial bonding strength than that of pure HA coatings. Osteoblasts were cultured on the coatings to evaluate the biocompatibility of coatings. The results of the Alamar blue and ALP test indicated that the composite coatings could favor the proliferation and differentiation of the osteoblasts, which indicated that the as-prepared HA/ZrO2 nanocomposite coatings had good biocompatibility.
In summary, three types of HA-contained composite coatings, HA/CS, HA/geltin and HA/ ZrO2, have been successfully produced on titanium substrates. The results of present study provided valuable reference for the further research in developing composite coatings for biomedical applications.
引文
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