Synthesis And Biocompatibility Of Spherical Nano-hydroxyapatite Bioluminescence Probe Materials

Hydroxyapatite?HA?is the main inorganic component of human teeth and animal bones.It has become the most studied inorganic material,and also been one of the most widely used biological materials because of its better biological compatibility and biological activity.The size,morphology,surface,biocompatibility and other aspects of synthetic biomaterials has important effects on the applications of the materials.Based on the principle of liquid phase method of preparation of HA,it was prepared by hydrothermal method.Above all the common compound amino acids of human body was adopted as the template to improve the morphology,size,biocompatibility of the nano HA.In order to detect the role of nano HA in organism process,rare earth was doped into nano HA.It is expected to make hydroxyapatite material become a type of new biological probe.The main content of this topic and main points were as follows:?1?The preparation of spherical nano HA.Hydrothermal method was applied in this experiment with calcium nitrate and diammonium hydrogen phosphate as raw materials,and the ratio Ca/P was controlled 10/6.With 20 kinds of L-amino acid as the template,spherical nano HA were synthesized under suitable condition of temperature,pH and reaction time.The structure and morphology of the prepared HA that was characterized via X-ray powder diffraction?XRD?and transmission electron microscope?TEM?.Results showed that L-arginine?pH=9,185??,L-threonine?pH=9,185??,L-histidine?pH=9,125??,L-tyrosine?pH=10,195??,L-asparagine?pH=10,185??,L-lysine?pH=11,175??,L-glutamic acid?pH=10,175??,L-alanine?pH=11,150??,L-leucine?pH=11,175??,L-methionine?pH=9,180??,L-tryptophan?pH=10,150??as the template and under the corresponding conditions of pH and temperature,the spherical nano HA were obtained with 30⁵0 nm of particle size,and that dispersion is good.?2?The preparation of spherical nano Tb-HA with same mole fraction of Tb³⁺.Based on the preparation of spherical nano HA with 11 kinds of L-amino acid as the template,2mol%of Tb³⁺was doped to prepare the nano Tb-HA under the certain condition of temperature and pH.Experimental results showed that,6 kinds of L-amino acids included L-glutamic acid?pH=10?,L-asparagine?pH=11?,L-methionine?pH=11?,L-isoleucine?pH=11?,L-alanine?pH=11?,L-tyrosine?pH=10?as template was demonstrated once again to prepare the spherical nano Tb-HA.?3?The preparation of spherical nano Tb-HA with different mole fraction of Tb³⁺and Eu³⁺.Based on the preparation of Tb-HA nanoparticles with with same mole fraction of Tb³⁺,L-asparagine,L-glutamic acid,L-methionine were used as templates respectively to prepare the nano Tb-HA,and doped with rare earth mole fraction of 0mol%,1 mol%,3 mol%,5 mol%,7 mol%.The characterization of structure morphology and luminous performance of crystal found that different L-amino acids as a template,the particle size and crystalline of Tb-HA would be different.Considering the crystalline of Tb-HA,which L-asparagine as template to prepare the spherical morphology of HA could doped with the most mole fraction was 3 mol%.L-methionine with that was is also 3 mol%and L-glutamic acid with that was 7mol%.?4?The luminescence property of Tb-HA and Eu-HA nanoparticles.Based on successful preparation of Tb-HA and Eu-HA doped with different mole fraction rare earth ion,the results about characterization of fluorescence spectrum showed that fluorescence performance of HA is good,and with the increase of doped molar fraction,fluorescence intensity gradually increased and the size of particle decreases slightly.Tb-HA excitied by 374 nm uv light could produce three characteristic fluorescence emission peak at 490nm of ⁵D₄-⁷F₆,544nm of ⁵D₄-⁷F₅ and 585nm of⁵D₄-⁷F₄.543 nm is the maximum emission wavelength.Eu-HA nanoparticles excitied by 394 nm uv light could produce two characteristic emission peaks at 592 nm of⁵D₀-⁷F₁ and 617 nm of ⁵D₀-⁷F₂.617 nm is the maximum emission wavelength.?5?Five sample with different Tb³⁺doping of mole fraction and two different morphology of Tb-HA were determined by MTT test.The method is to explore the influence of sample by co-culture with mouse fibroblast cells?L929 cells?and the concentration of the sample is 500?g/mL,1000?g/mL,1500?g/mL and 2000?g/mL.The experiment results showed that increase with the concentration of the sample,the cell toxicity is bigger and cultivate the longer of cocultivation,the cytotoxicity is bigger.When the samples were co-cultured for 24h,it was non-toxic.When it was developed to 48h,slightly toxic was found.If it cultured over 72h,the cytotoxicity would be obvious.It is recommended that the training time is no more than 48 h.