激光快速成形技术制作纯钛植入材料的实验研究
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摘要
LRF技术与传统金属加工技术相比具有显著的优势。为了详细了解LRF技术用于纯钛植入材料制作的可行性,本研究利用LRF技术制作了纯钛植入材料,并对LRF Ti的物理机械性能、生物安全性、生物相容性和耐腐蚀性进行了评价。
第Ⅰ部分LRF Ti的制备和相关机械性能评价
选用合适的工艺参数制作LRF Ti材料,选用的激光成形工艺参数为:激光功率:2700W;光斑直径:3mm;光束扫描速度:500mm/min;保护气体是氩气:载粉气流量250L/h,约束气流量5～7.5L/min,送粉速度4.2 g/min。采用多道搭接Z向生长方式成形,成形过程层高0.5 mm。然后根据国家标准测试LRF Ti材料的机械性能、显微硬度和疲劳强度,并对材料的化学成分和组织结构进行观察和分析,对LRF Ti用作植入材料的生物力学相容性进行评价。
结果:按照国家标准GB/T 228-2002进行拉伸实验,LRF Ti试样的抗拉强度(475 MPa)、屈服强度(383MPa)均高于GB/T 13810-1997中TA2纯钛板材的标准(440MPa、320 MPa),特别是延伸率,LRF Ti试样达到27%,远高于医用TA2延伸率标准中18%的规定。试样的氧含量(0.083%)低于YY0315-1999标准中TA1、TA2的氧含量(0.15%、0.25%)。从拉伸断面扫描电镜照片可以看出断面主要为大小不一的韧窝,可以判断为典型的韧性断裂。杨氏模量(103.7GPa)稍低于TA2的弹性模量(108 Gpa)。扫描电镜(SEM)微观组织照片显示LRF Ti的微观组织结构均匀,无气孔、裂纹等缺陷;α和β相相互交织成网篮状组织。显微硬度值(188.4～206.3)稍高于CP Ti(TA2)的显微硬度值(166.3～181.9)。LRF Ti在室温下循环107次时的疲劳强度极限是250-275MPa,高于TA1的193 MPa和TA2的230 MPa。
结论:利用纯钛粉末通过LRF技术制备了规格为90mm×30mm×15mm、外表规则光滑、亮白色、低氧含量(0.083%)的LRF Ti试样。LRF Ti试样具有更好的生物力学相容性,机械性能满足牙科植入物用纯钛材的要求,符合生物医学硬组织替代材料的标准。
第Ⅱ部分LRF Ti的生物安全性评价
本研究按照标准GB/T 16886.5-2002、YY/T 0279-1995、YY/T 0127.1-93、YY/T 0127.2-93,分别从细胞毒性试验、口腔粘膜刺激试验、溶血试验、急性全身毒性试验和骨内植入试验等方面,综合评价LRF Ti的生物安全性。
结果:LRF Ti的细胞毒性测试结果为0级,材料缝合处动物口腔颊囊粘膜无病理反应,即对粘膜无刺激反应。溶血率为2.68%,小于5%,不会引起急性溶血。将LRF Ti材料浸提液注射入动物体内,每组动物在不同时间段的体重变化无显著性差异(P>0.05),在72h的观察期内,无急性毒性症状出现。骨内埋植试验结果可见种植体与骨组织接触紧密,周围骨组织无吸收现象。种植体表面有骨陷窝和骨组织形成,周围骨组织内的成骨细胞和成纤维细胞的细胞突向种植体表面伸出,LRF Ti和CP Ti种植体没有明显差别。
结论:LRF Ti具有优良的生物安全性。
第Ⅲ部分LRF Ti的生物相容性评价
通过比较LRF Ti和CP Ti表面的接触角和表面能、小鼠颅骨成骨细胞在试样表面的附着动力学、成骨细胞碱性磷酸酶活性以及细胞在试样表面的形态,评价LRF Ti材料的生物相容性。
结果表明:LRF Ti的表面能大于CP Ti。在成骨细胞培养的初期,两者的细胞增殖速度无差异;随着细胞培养时间的延长,LRF Ti组的成骨细胞增殖速度快于CP Ti组,碱性磷酸酶的活性增加;细胞形态显示细胞功能活跃,活性增加。
结论:LRF Ti能够刺激成骨细胞的增值生长,具有良好的生物相容性。第Ⅳ部分LRF Ti的耐腐蚀性能评价
根据国家标准,通过对比观察LRF Ti和医用钛及钛合金在不同溶液中的金属离子析出情况、应力加载下的耐腐蚀情况,以及氟离子和pH值对电化学行为的影响等,评价LRF Ti材料的耐腐蚀性能。
结果:在人工唾液、乳酸、2%NaF人工唾液(PH=7)和2%NaF人工唾液(PH=3)中,LRF Ti的钛离子析出量分别是62ng/ml、302ng/ml、24.7μg/ml和156.7μg/ml,低于CP Ti的66.1ng/ml、353ng/ml、33.1μg/ml和295.7μg/ml。氟与钛表面的氧化膜反应形成腐蚀产物Na3TiF6;在2%NaF人工唾液(PH=3)中,钛离子的析出量最大。在应力和氟离子的共同作用下,CP Ti的腐蚀程度比LRF Ti严重。在人工唾液、2%NaF人工唾液(PH=7)和2%NaF人工唾液(PH=3)中,LRF Ti的自腐蚀电位分别为-469mV、-925mV和-943mV,均高于CP Ti的-555mV、-943mV和-956mV;自腐蚀电流密度分别为2.14×10-6 Acm-2、2.65×10-6 Acm-2和4.51×10-6 Acm-2 ,均小于CP Ti的2.18×10-6Acm-2、4.49×10-6Acm-2和6.63×10-6Acm-2;膜电阻(2.77×106?cm2、3.41×105?cm2和4.3×104?cm2)和传递电阻(6.71×104?cm2、2028?cm2和4.4×103?cm2)分别高于CP Ti的膜电阻(1.42×106?cm2、6.65×104?cm2和16.93?cm2)和传递电阻(236.9?cm2、44.1?cm2和2.56×103?cm2)。
结论:pH值的降低和溶液中氟离子的存在能够加速钛离子的释放。在人工唾液和含氟溶液中LRF Ti耐电化学腐蚀和耐应力腐蚀性能比CP Ti好。
To investigate the feasibility of laser rapid forming titanium(LRF Ti)for medical implant materials,this paper evaluated the mechanical properties, biological safety,biocompatibility and corrosion behavior of LRF Ti according to national standard.
PartⅠFabrication of pure titanium specimens by LRF technique and the test of mechanical properties
Pure Ti specimens were fabricated by LRF technique with suitable parameters.The parameters used in the test were:power,2200W;spot diameter, 3mm.The powder was delivered in a gas jet through nozzles coaxially with the laser beam.Powder feeder rate was 4.2g/min.The process was occurred in closed chambers with controlled inert atmospheres(Ar:250L/h, 5-7.5L/min).The layer thickness was 0.5mm.To evaluate the mechanical properties of LRF Ti,the specimens underwent tensile test,microhardness test,chemical composition analysis,microstructure observation and fatigue test.
Results:The tensile strength and yield strength of LRF Ti were 475 MPa and 383MPa,they were higher than TA2 respectively according to GB/T 13810-1997.The elongation(27%) was far higher than TA2(18%). The oxygen content of LRF Ti was 0.083% and far below TA1(0.15%) and TA2(0.25%) according to YY0315-1999.The fracture surface of tensile specimens was ductile. Yang’s modulus of LRF Ti(103.7GPa) was slightly lower than TA2 (108GPa).The microstructure of LRF Ti was basket -weave structure. The Vickers hardness is 188.4~206.3.The fatigue strength of LRF Ti was 250-275MPa at the end of 107cycles while TA1 and TA2 were 193 and 230 MPa respectively.
Conclusion: The LRF Ti specimens fabricated completely were silvery with a smooth surface with low oxygen content. The LRF Ti had better mechanical compatibility and the mechanical properties meet the requirement of pure titanium implant materials.
PartⅡBiological safety test of LRF Ti
Biological tests were carried out to evaluate the biological safety of LRF Ti for biomedical materials according to standards GB/T 16886.5-2002,YY/T 0279-1995,YY/T 0127.1-93 and YY/T 0127.2-93.The biological tests included Cytotoxicity test, oral mucous membrane irritation test ,hemolysis test, acute systemic toxicity test and bone implant test.
Results: The cytotoxicity rank of LRF Ti was 0; The oral mucous membrane surround LRF Ti specimens had no pathological reaction and membrane irritation. Hemolysis ratio was 2.68% and fell bellow the standard limits (5% ,YY/T 0127.1-93),LRF Ti didn’t result in hemolysis. The animals had no signs of toxicity during 72 hour after injected with the leaching liquor of LRF Ti,the weights of animals had no statistical difference between different test periods compared to controlled group. The animals had no bone absorption near the LRF Ti implants , the implants contacted bone tissue tightly. There were bone lacunas and new osseous tissue formation on the surface of LRF Ti implants. The synapsis of osteoblasts and fibroblasts stretched to the surface of LRF Ti implants. The reacton of bone tissue surrounded LRF Ti implants had no difference with CP Ti implants in vivo.
Conclusion: LRF Ti had excellent biological safety and meet the requirement of biological test standard.
PartⅢBiocompatibility test of LRF Ti
Cell culture tests in vitro were undertook to evaluate the biocompatibility of LRF Ti. The cell cranium osteoblasts of mouse were used in the experiment. The contact angles and total surface energies of LRF Ti and CP Ti specimens were tested also.
Results: The total surface energy of LRF Ti sample was higher than CP Ti sample. The cell growth rate had no statistical difference between LRF Ti and CP Ti samples. The multiplication of LRF Ti specimen was faster than CP Ti specimen with the cultivation time increased. The alkaline phosphatase (ALP) activity were increased consequently. The morphology of osteoblast indicated high activity of osteoblast on the surface of LRF Ti specimen.
Conclusion: LRF Ti stimulated osteoblasts multiplication and had favorable biocompatibility.
PartⅣThe corrosion behavior of LRF Ti
Electrochemical test,static immersion test and stress-corrosion test were used to evaluate the corrosion behavior of LRF Ti according to standard. The influence of fluoride and pH value on the corrosion resistance of LRF Ti were also investigated.
Results:In artificial saliva,1%lactic acid solution,2%NaF artificial saliva (pH=7) and 2%NaF artificial saliva (pH=3),the amounts of Ti ion released from LRF Ti sample(62ng/ml,302ng/ml,24.7μg/ml and 156.7μg/ml) were lower than CP Ti (66.1ng/ml, 353ng/ml, 33.1μg/ml and 295.7μg/ml) respectively;The corrosion products were Na3TiF6. The greatest amounts of metal ion release were observed in 2% NaF artificial saliva (pH=3). LRF Ti had less corrosion products and pitting corrosion than CP Ti in stress-corrosion test. In artificial saliva, 2%NaF artificial saliva(pH=7) and 2%NaF artificial saliva (pH=3),the free circuit potential (Ecorr) of LRF Ti (-469mV,-925mV and-943mV) were higher than those of CP Ti (-555mV,-943mV and-956mV);the current density of LRF Ti (2.14×10-6 Acm-2、2.65×10-6 Acm-2 and 4.51×10-6 Acm-2) were lower than those of CP Ti (2.18×10-6Acm-2、4.49×10-6Acm-2 and 6.63×10-6Acm-2),the film resistance (2.77×106?cm2、3.41×105?cm2 and 4.3×104?cm2)and transfer resistance (6.71×104?cm2、2028?cm2 and 4.4×103?cm2)of LRF Ti were higher than those of CP Ti(1.42×106?cm2、6.65×104?cm2and16.93?cm2)(236.9?cm2、44.1 ?cm2and 2.56×103?cm2)respectively。
Conclusion: Low pH value and fluoride in the solution could accelerate Ti iron release,LRF Ti had better corrosion resistance for long-term implants than CP Ti.
第Ⅰ部分LRF Ti的制备和相关机械性能评价
选用合适的工艺参数制作LRF Ti材料,选用的激光成形工艺参数为:激光功率:2700W;光斑直径:3mm;光束扫描速度:500mm/min;保护气体是氩气:载粉气流量250L/h,约束气流量5～7.5L/min,送粉速度4.2 g/min。采用多道搭接Z向生长方式成形,成形过程层高0.5 mm。然后根据国家标准测试LRF Ti材料的机械性能、显微硬度和疲劳强度,并对材料的化学成分和组织结构进行观察和分析,对LRF Ti用作植入材料的生物力学相容性进行评价。
结果:按照国家标准GB/T 228-2002进行拉伸实验,LRF Ti试样的抗拉强度(475 MPa)、屈服强度(383MPa)均高于GB/T 13810-1997中TA2纯钛板材的标准(440MPa、320 MPa),特别是延伸率,LRF Ti试样达到27%,远高于医用TA2延伸率标准中18%的规定。试样的氧含量(0.083%)低于YY0315-1999标准中TA1、TA2的氧含量(0.15%、0.25%)。从拉伸断面扫描电镜照片可以看出断面主要为大小不一的韧窝,可以判断为典型的韧性断裂。杨氏模量(103.7GPa)稍低于TA2的弹性模量(108 Gpa)。扫描电镜(SEM)微观组织照片显示LRF Ti的微观组织结构均匀,无气孔、裂纹等缺陷;α和β相相互交织成网篮状组织。显微硬度值(188.4～206.3)稍高于CP Ti(TA2)的显微硬度值(166.3～181.9)。LRF Ti在室温下循环107次时的疲劳强度极限是250-275MPa,高于TA1的193 MPa和TA2的230 MPa。
结论:利用纯钛粉末通过LRF技术制备了规格为90mm×30mm×15mm、外表规则光滑、亮白色、低氧含量(0.083%)的LRF Ti试样。LRF Ti试样具有更好的生物力学相容性,机械性能满足牙科植入物用纯钛材的要求,符合生物医学硬组织替代材料的标准。
第Ⅱ部分LRF Ti的生物安全性评价
本研究按照标准GB/T 16886.5-2002、YY/T 0279-1995、YY/T 0127.1-93、YY/T 0127.2-93,分别从细胞毒性试验、口腔粘膜刺激试验、溶血试验、急性全身毒性试验和骨内植入试验等方面,综合评价LRF Ti的生物安全性。
结果:LRF Ti的细胞毒性测试结果为0级,材料缝合处动物口腔颊囊粘膜无病理反应,即对粘膜无刺激反应。溶血率为2.68%,小于5%,不会引起急性溶血。将LRF Ti材料浸提液注射入动物体内,每组动物在不同时间段的体重变化无显著性差异(P>0.05),在72h的观察期内,无急性毒性症状出现。骨内埋植试验结果可见种植体与骨组织接触紧密,周围骨组织无吸收现象。种植体表面有骨陷窝和骨组织形成,周围骨组织内的成骨细胞和成纤维细胞的细胞突向种植体表面伸出,LRF Ti和CP Ti种植体没有明显差别。
结论:LRF Ti具有优良的生物安全性。
第Ⅲ部分LRF Ti的生物相容性评价
通过比较LRF Ti和CP Ti表面的接触角和表面能、小鼠颅骨成骨细胞在试样表面的附着动力学、成骨细胞碱性磷酸酶活性以及细胞在试样表面的形态,评价LRF Ti材料的生物相容性。
结果表明:LRF Ti的表面能大于CP Ti。在成骨细胞培养的初期,两者的细胞增殖速度无差异;随着细胞培养时间的延长,LRF Ti组的成骨细胞增殖速度快于CP Ti组,碱性磷酸酶的活性增加;细胞形态显示细胞功能活跃,活性增加。
结论:LRF Ti能够刺激成骨细胞的增值生长,具有良好的生物相容性。第Ⅳ部分LRF Ti的耐腐蚀性能评价
根据国家标准,通过对比观察LRF Ti和医用钛及钛合金在不同溶液中的金属离子析出情况、应力加载下的耐腐蚀情况,以及氟离子和pH值对电化学行为的影响等,评价LRF Ti材料的耐腐蚀性能。
结果:在人工唾液、乳酸、2%NaF人工唾液(PH=7)和2%NaF人工唾液(PH=3)中,LRF Ti的钛离子析出量分别是62ng/ml、302ng/ml、24.7μg/ml和156.7μg/ml,低于CP Ti的66.1ng/ml、353ng/ml、33.1μg/ml和295.7μg/ml。氟与钛表面的氧化膜反应形成腐蚀产物Na3TiF6;在2%NaF人工唾液(PH=3)中,钛离子的析出量最大。在应力和氟离子的共同作用下,CP Ti的腐蚀程度比LRF Ti严重。在人工唾液、2%NaF人工唾液(PH=7)和2%NaF人工唾液(PH=3)中,LRF Ti的自腐蚀电位分别为-469mV、-925mV和-943mV,均高于CP Ti的-555mV、-943mV和-956mV;自腐蚀电流密度分别为2.14×10-6 Acm-2、2.65×10-6 Acm-2和4.51×10-6 Acm-2 ,均小于CP Ti的2.18×10-6Acm-2、4.49×10-6Acm-2和6.63×10-6Acm-2;膜电阻(2.77×106?cm2、3.41×105?cm2和4.3×104?cm2)和传递电阻(6.71×104?cm2、2028?cm2和4.4×103?cm2)分别高于CP Ti的膜电阻(1.42×106?cm2、6.65×104?cm2和16.93?cm2)和传递电阻(236.9?cm2、44.1?cm2和2.56×103?cm2)。
结论:pH值的降低和溶液中氟离子的存在能够加速钛离子的释放。在人工唾液和含氟溶液中LRF Ti耐电化学腐蚀和耐应力腐蚀性能比CP Ti好。
To investigate the feasibility of laser rapid forming titanium(LRF Ti)for medical implant materials,this paper evaluated the mechanical properties, biological safety,biocompatibility and corrosion behavior of LRF Ti according to national standard.
PartⅠFabrication of pure titanium specimens by LRF technique and the test of mechanical properties
Pure Ti specimens were fabricated by LRF technique with suitable parameters.The parameters used in the test were:power,2200W;spot diameter, 3mm.The powder was delivered in a gas jet through nozzles coaxially with the laser beam.Powder feeder rate was 4.2g/min.The process was occurred in closed chambers with controlled inert atmospheres(Ar:250L/h, 5-7.5L/min).The layer thickness was 0.5mm.To evaluate the mechanical properties of LRF Ti,the specimens underwent tensile test,microhardness test,chemical composition analysis,microstructure observation and fatigue test.
Results:The tensile strength and yield strength of LRF Ti were 475 MPa and 383MPa,they were higher than TA2 respectively according to GB/T 13810-1997.The elongation(27%) was far higher than TA2(18%). The oxygen content of LRF Ti was 0.083% and far below TA1(0.15%) and TA2(0.25%) according to YY0315-1999.The fracture surface of tensile specimens was ductile. Yang’s modulus of LRF Ti(103.7GPa) was slightly lower than TA2 (108GPa).The microstructure of LRF Ti was basket -weave structure. The Vickers hardness is 188.4~206.3.The fatigue strength of LRF Ti was 250-275MPa at the end of 107cycles while TA1 and TA2 were 193 and 230 MPa respectively.
Conclusion: The LRF Ti specimens fabricated completely were silvery with a smooth surface with low oxygen content. The LRF Ti had better mechanical compatibility and the mechanical properties meet the requirement of pure titanium implant materials.
PartⅡBiological safety test of LRF Ti
Biological tests were carried out to evaluate the biological safety of LRF Ti for biomedical materials according to standards GB/T 16886.5-2002,YY/T 0279-1995,YY/T 0127.1-93 and YY/T 0127.2-93.The biological tests included Cytotoxicity test, oral mucous membrane irritation test ,hemolysis test, acute systemic toxicity test and bone implant test.
Results: The cytotoxicity rank of LRF Ti was 0; The oral mucous membrane surround LRF Ti specimens had no pathological reaction and membrane irritation. Hemolysis ratio was 2.68% and fell bellow the standard limits (5% ,YY/T 0127.1-93),LRF Ti didn’t result in hemolysis. The animals had no signs of toxicity during 72 hour after injected with the leaching liquor of LRF Ti,the weights of animals had no statistical difference between different test periods compared to controlled group. The animals had no bone absorption near the LRF Ti implants , the implants contacted bone tissue tightly. There were bone lacunas and new osseous tissue formation on the surface of LRF Ti implants. The synapsis of osteoblasts and fibroblasts stretched to the surface of LRF Ti implants. The reacton of bone tissue surrounded LRF Ti implants had no difference with CP Ti implants in vivo.
Conclusion: LRF Ti had excellent biological safety and meet the requirement of biological test standard.
PartⅢBiocompatibility test of LRF Ti
Cell culture tests in vitro were undertook to evaluate the biocompatibility of LRF Ti. The cell cranium osteoblasts of mouse were used in the experiment. The contact angles and total surface energies of LRF Ti and CP Ti specimens were tested also.
Results: The total surface energy of LRF Ti sample was higher than CP Ti sample. The cell growth rate had no statistical difference between LRF Ti and CP Ti samples. The multiplication of LRF Ti specimen was faster than CP Ti specimen with the cultivation time increased. The alkaline phosphatase (ALP) activity were increased consequently. The morphology of osteoblast indicated high activity of osteoblast on the surface of LRF Ti specimen.
Conclusion: LRF Ti stimulated osteoblasts multiplication and had favorable biocompatibility.
PartⅣThe corrosion behavior of LRF Ti
Electrochemical test,static immersion test and stress-corrosion test were used to evaluate the corrosion behavior of LRF Ti according to standard. The influence of fluoride and pH value on the corrosion resistance of LRF Ti were also investigated.
Results:In artificial saliva,1%lactic acid solution,2%NaF artificial saliva (pH=7) and 2%NaF artificial saliva (pH=3),the amounts of Ti ion released from LRF Ti sample(62ng/ml,302ng/ml,24.7μg/ml and 156.7μg/ml) were lower than CP Ti (66.1ng/ml, 353ng/ml, 33.1μg/ml and 295.7μg/ml) respectively;The corrosion products were Na3TiF6. The greatest amounts of metal ion release were observed in 2% NaF artificial saliva (pH=3). LRF Ti had less corrosion products and pitting corrosion than CP Ti in stress-corrosion test. In artificial saliva, 2%NaF artificial saliva(pH=7) and 2%NaF artificial saliva (pH=3),the free circuit potential (Ecorr) of LRF Ti (-469mV,-925mV and-943mV) were higher than those of CP Ti (-555mV,-943mV and-956mV);the current density of LRF Ti (2.14×10-6 Acm-2、2.65×10-6 Acm-2 and 4.51×10-6 Acm-2) were lower than those of CP Ti (2.18×10-6Acm-2、4.49×10-6Acm-2 and 6.63×10-6Acm-2),the film resistance (2.77×106?cm2、3.41×105?cm2 and 4.3×104?cm2)and transfer resistance (6.71×104?cm2、2028?cm2 and 4.4×103?cm2)of LRF Ti were higher than those of CP Ti(1.42×106?cm2、6.65×104?cm2and16.93?cm2)(236.9?cm2、44.1 ?cm2and 2.56×103?cm2)respectively。
Conclusion: Low pH value and fluoride in the solution could accelerate Ti iron release,LRF Ti had better corrosion resistance for long-term implants than CP Ti.
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