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Synthesis and Characterization of Amorphous Calcium Phosphate Nanoparticles

비정질 칼슘 포스페이트 나노 입자의 합성과 특성

  • Han, Ji-Hoon (Department of Polymer Science and Chemical Engineering, Pusan National University) ;
  • Chung, Sungwook (School of Chemical, Biomolecular, and Environmental Engineering, Pusan National University)
  • 한지훈 (부산대학교 화학공학.고분자공학과) ;
  • 정성욱 (부산대학교 화공생명.환경공학부)
  • Received : 2018.08.23
  • Accepted : 2018.09.17
  • Published : 2018.12.10

Abstract

The synthesis and characterization of amorphous calcium phosphate (ACP) nanoparticles were reported in this work. We show that relatively monodisperse ACP nanoparticles with a size of sub-100 nm can be prepared by a hydrothermal reaction of calcium chloride ($CaCl_2$) and disodium adenosine triphosphate ($Na_2ATP$) in the presence of sodium phytate as an additive. Their compositions and structures were confirmed using a series of material characterization techniques. Our results exhibit that ACP nanoparticles synthesized using sodium phytate enhanced the stability of maintaining their amorphous nature and prevented from a conversion to crystalline hydroxyapatite (HAP). ACP nanoparticles with the improved stability have potential uses in biomaterial applications in regenerative medicine.

본 연구에서는 비정질 칼슘 포스페이트(ACP) 나노 입자의 합성과 특성 분석을 진행하였다. 염화칼슘(calcium chloride ($CaCl_2$))과 아데노신 인산나트륨(disodium adenosine triphosphate ($Na_2ATP$)) 그리고 피트산 나트륨(sodium phytate) 첨가제를 열수 반응을 통해 상대적으로 단분산된 100 nm 크기 이하의 ACP 나노 입자를 성공적으로 합성하였고 나노 입자의 화학적 조성과 구조를 재료 분석을 통해 확인하였다. 피트산 나트륨 첨가제의 사용을 통해 얻은 ACP 나노 입자는 비정질성을 유지하고 결정성 하이드록시아파타이트(HAP)로의 전환을 방지하는 안정성이 향상되었음을 발견하였다. 본 연구를 통해 발견된 향상된 안정성을 가지는 ACP 나노 입자는 재생 의학 분야에서의 생체 적합 물질로의 응용에 중요한 잠재적 용도가 있을 것이라 사료된다.

Keywords

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Figure 1. Schematic illustration of the preparation of amorphous calcium phosphate (ACP) nanoparticles via a hydrothermal method.

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Figure 2. FESEM images of as-synthesized ACP nanoparticles. (A) ACP nanoparticles synthesized in the presence of MgCl2. Inset is a higher magnification FESEM image. (B) ACP nanoparticles synthesized in the presence of sodium phytate. (C) Crystalline hydroxyapatite (HAP) nanorods that were transformed from ACP nanoparticles synthesized with no additives after a short induction period. Inset is a higher magnification FESEM image.

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Figure 3. FESEM images and their corresponding EDS spectra of as-synthesized ACP nanoparticles. (A) ACP nanoparticles synthesized with no additives. (B) ACP nanoparticles synthesized in the presence of MgCl2.

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Figure 4. X-ray powder diffraction (XRD) patterns of ACP nanoparticles and HAP nanorods. Blue curve (ACP1) exhibits the XRD pattern of the ACP nanoparticles synthesized in the presence of MgCl2 (blue curve). Red curve (ACP2) shows the XRD pattern of the ACP particles synthesized in the presence of sodium phytate. Green curve (HAP) corresponding to the XRD pattern of HAP nanorods shows their characteristic peaks (denoted with asterisk marks) that are assigned based on JCPDS No. 09-0432 (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article).

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Figure 5. FT-IR spectra of as-synthesized ACP nanoparticles and HAP nanorods. Black curve corresponds to FT-IR of Na2ATP. Red (ACP1) and blue (ACP2) curves correspond to the spectra from the as-synthesized ACP nanoparticles in the presence of MgCl2 and sodium phytate, respectively. Green (HAP) curve corresponds to HAP nanorods that were transformed from ACP nanoparticles synthesized with no additive. The positions of characteristic ν3 and ν4 vibrations of PO43- groups were embodied with the arrows in each spectrum (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article).

References

  1. C. Qi, J. Lin, L. H. Fu, and P. Huang, Calcium-based biomaterials for diagnosis, treatment, and theranostics, Chem. Soc. Rev., 47, 357-403 (2018). https://doi.org/10.1039/C6CS00746E
  2. C. Combes, S. Cazalbou, and C. Rey, Apatite biominerals, Minerals, 6, 1-25 (2016).
  3. W. J. Jin, S. Q. Jiang, H. H. Pan, and R. K. Tang, Amorphous phase mediated crystallization: Fundamentals of biomineralization, Crystals, 8, 1-24 (2018).
  4. H. R. Wang et al., Oriented and ordered biomimetic remineralization of the surface of demineralized dental enamel using HAP@ACP nanoparticles guided by glycine, Sci. Rep., 7, 40701-40713 (2017). https://doi.org/10.1038/srep40701
  5. E. Beniash, R. A. Metzler, R. S. K. Lam, and P. U. P. A. Gilbert, Transient amorphous calcium phosphate in forming enamel, J. Struct. Biol., 166, 133-143 (2009). https://doi.org/10.1016/j.jsb.2009.02.001
  6. A. Dey et al., The role of prenucleation clusters in surface-induced calcium phosphate crystallization, Nat. Mater., 9, 1010-1014 (2010). https://doi.org/10.1038/nmat2900
  7. H. Zhou and J. Lee, Nanoscale hydroxyapatite particles for bone tissue engineering, Acta Biomater., 7, 2769-2781 (2011). https://doi.org/10.1016/j.actbio.2011.03.019
  8. M. Nagano, T. Nakamura, T. Kokubo, M. Tanahashi, and M. Ogawa, Differences of bone bonding ability and degradation behavior in vivo between amorphous calcium phosphate and highly crystalline hydroxyapatite coating, Biomaterials, 17, 1771-1777 (1996). https://doi.org/10.1016/0142-9612(95)00357-6
  9. A. L. Boskey, Amorphous calcium phosphate: The contention of bone, J. Dent. Res., 76, 1433-1436 (1997). https://doi.org/10.1177/00220345970760080501
  10. S. Kim, H. S. Ryu, H. Shin, H. S. Jung, and K. S. Hong, In situ observation of hydroxyapatite nanocrystal formation from amorphous calcium phosphate in calcium-rich solutions, Mater. Chem. Phys., 91, 500-506 (2005). https://doi.org/10.1016/j.matchemphys.2004.12.016
  11. C. G. Wang et al., Crystallization at Multiple Sites inside Particles of Amorphous Calcium Phosphate, Cryst. Growth Des., 9, 2620-2626 (2009). https://doi.org/10.1021/cg801069t
  12. A. L. Boskey and A. S. Posner, Conversion of amorphous calcium phosphate to microcrystalline hydroxyapatite. A pH-dependent, solution- mediated, solid-solid conversion, J. Phys. Chem., 77, 2313-2317 (1973). https://doi.org/10.1021/j100638a011
  13. G. H. Nancollas and B. Tomazic, Growth of calcium-phosphate on hydroxyapatite crystals - Effect of supersaturation and ionic medium, J. Phys. Chem., 78, 2218-2225 (1974). https://doi.org/10.1021/j100615a007
  14. H. C. Margolis, S. Y. Kwak, and H. Yamazaki, Role of mineralization inhibitors in the regulation of hard tissue biomineralization: Relevance to initial enamel formation and maturation, Front. Physiol., 5:339 (2014).
  15. S. Q. Jiang, W. Jin, Y.-N. Wang, H. Pan, Z. Sun, and R. Tang, Effect of the aggregation state of amorphous calcium phosphate on hydroxyapatite nucleation kinetics, RSC Adv., 7, 25497-25503 (2017). https://doi.org/10.1039/C7RA02208E
  16. Z. Zyman, D. Rokhmistrov, and V. Glushko, Structural changes in precipitates and cell model for the conversion of amorphous calcium phosphate to hydroxyapatite during the initial stage of precipitation, J. Cryst. Growth, 353, 5-11 (2012). https://doi.org/10.1016/j.jcrysgro.2012.04.041
  17. H. Furedi-Milhofer, L. Brecevic, and B. Purgaric, Crystal growth and phase transformation in the precipitation of calcium phosphates, Faraday Discuss. Chem. Soc., 61, 184-193 (1976). https://doi.org/10.1039/DC9766100184
  18. S. Q. Jiang, H. H. Pan, Y. Chen, X. R. Xu, and R. K. Tang, Amorphous calcium phosphate phase-mediated crystal nucleation kinetics and pathway, Faraday Discuss., 179, 451-461 (2015). https://doi.org/10.1039/C4FD00212A
  19. R. Wuthier and E. Eanes, Effect of phospholipids on the transformation of amorphous calcium phosphate to hydroxyapatite in vitro, Calcif. Tissue Res., 19, 197-210 (1975). https://doi.org/10.1007/BF02564004
  20. R. Z. LeGeros et al., Amorphous calcium phosphates (ACP): Formation and stability, Key Eng. Mater., 284, 7-10 (2005).
  21. N. C. Blumenthal, F. Betts, and A. S. Posner, Stabilization of amorphous calcium-phosphate by Mg and ATP, Calcif. Tissue Res., 23, 245-250 (1977). https://doi.org/10.1007/BF02012793
  22. Y. Chen, W. J. Gu, H. H. Pan, S. Q. Jiang, and R. K. Tang, Stabilizing amorphous calcium phosphate phase by citrate adsorption, Cryst. Eng. Comm., 16, 1864-1867 (2014). https://doi.org/10.1039/C3CE42274G
  23. Z. Amjad, Inhibition of the amorphous calcium phosphate phase transformation reaction by polymeric and non-polymeric inhibitors, Phosphorus Res. Bull., 7, 45-54 (1997). https://doi.org/10.3363/prb1992.7.0_45
  24. C. Qi, Y.-J. Zhu, X.-Y. Zhao, B.-Q. Lu, Q.-L. Tang, J. Zhao, and F. Chen, Highly stable amorphous calcium phosphate porous nanospheres: Microwave-assisted rapid synthesis using ATP as phosphorus source and stabilizer, and their application in anticancer drug delivery, Chemistry, 19, 981-987 (2013). https://doi.org/10.1002/chem.201202829
  25. Y. Tanizawa and T. Suzuki, Effects of silicate ions on the formation and transformation of calcium phosphates in neutral aqueous solutions, J. Chem. Soc. Faraday Trans., 91, 3499-3503 (1995). https://doi.org/10.1039/ft9959103499
  26. P. Bar-Yosef Ofir, R. Govrin-Lippman, N. Garti, and H. Füredi-Milhofer, The influence of polyelectrolytes on the formation and phase transformation of amorphous calcium phosphate, Cryst. Growth Des., 4, 177-183 (2004). https://doi.org/10.1021/cg034148g
  27. M. J. Root, Inhibition of the amorphous calcium phosphate phase transformation reaction by polyphosphates and metal ions, Calcif. Tissue Int., 47, 112-116 (1990). https://doi.org/10.1007/BF02555994
  28. C. Qi, Q. L. Tang, Y. J. Zhu, X. Y. Zhao, and F. Chen, Microwave-assisted hydrothermal rapid synthesis of hydroxyapatite nanowires using adenosine 5'-triphosphate disodium salt as phosphorus source, Mater. Lett., 85, 71-73 (2012). https://doi.org/10.1016/j.matlet.2012.06.106
  29. F. Syberg, Y. Suveyzdis, C. Koetting, K. Gerwert, and E. Hofmann, Time-resolved Fourier transform infrared spectroscopy of the nucleotide-binding domain from the ATP-binding cassette transporter MsbA, J. Biol. Chem., 287, 23923-23931 (2012). https://doi.org/10.1074/jbc.M112.359208
  30. M. Liu, M. Krasteva, and A. Barth, Interaction of phosphate groups of ATP and aspartyl phosphate with the Sarcoplasmic Reticulum $Ca^{2+}$-ATPase: A FTIR study, Biophys. J., 89, 4352-4363 (2005). https://doi.org/10.1529/biophysj.105.061689
  31. Z. F. Zhou et al., Calcium phosphate-phosphorylated adenosine hybrid microspheres for anti-osteosarcoma drug delivery and osteogenic differentiation, Biomaterials, 121, 1-14 (2017). https://doi.org/10.1016/j.biomaterials.2016.12.031
  32. Z. He, C. W. Honeycutt, T. Zhang, and P. M. Bertsch, Preparation and FT-IR characterization of metal phytate compounds, J. Environ. Qual., 35, 1319-1328 (2006). https://doi.org/10.2134/jeq2006.0008

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