The Functionalization of Calcium Phosphate Materials of Protein-Based Biologically Active Molecules

Main Article Content

E.A. Kuvshinova
N.V. Petrakova
N.S. Sergeeva
V.A. Kirsanova
I.K. Sviridova
A.Y. Teterina
V.S. Komlev
A.D. Kaprin


Recent approaches to the calcium phosphate (CaP) materials functionalization with drugs and biomolecules have been actively developed for bone defect reconstruction. However, the current techniques are low efficient in context of drug incorporation and non-controlled release from the materials. Eventually, continuous therapeutic effect in bone defect area couldn’t be achieved. The aim of this work was to develop an effective method for biologically active molecules incorporation onto the surface of CаP materials, and to study the dynamics of its release. Octacalcium phosphate (OCP), β-tricalcium phosphate (β-TCP) and β-tricalcium phosphate with biomimetic calcium phosphate layer (β-TCPmod.) were used as ceramic bioactive carriers. Bovine serum albumin (BSA) was used as a model compounds. BSA incorporation on the ceramics surface was performed by biomimetic co-precipitation from several buffer solutions containing the incorporated compound. The efficiency of biomolecules incorporation was evaluated by measuring BSA concentrations in solutions before and after materials incubation. The release of the incorporated molecules from the materials was investigated for 6 days. The structure and composition of the obtained materials were studied by application of XRD, FTIR, SEM, BET methods. It was shown that the OCP specific surface (surface area, (SBET)) was almost in 12 times higher than SBET of β-TCP. By using biomimetic approach the increase of β-TCP surface area in 1.6 times was achieved; this enhanced protein incorporation more than 3 times. The BSA biomimetic co-precipitation together with CaP on the OCP surface proved to be more effective than its adsorption from salt free solutions. The study of BSA release revealed that only 45% of loaded albumin released during 6 days of observation. Therefore, the effective method of CaP functionalization was developed. Based on biomolecules incorporation by biomimetic co-precipitation from CaP solutions, it provided a low rate of its release.

Article Details

How to Cite
Kuvshinova, E., Petrakova, N., Sergeeva, N., Kirsanova, V., Sviridova, I., Teterina, A., Komlev, V., & Kaprin, A. (2019). The Functionalization of Calcium Phosphate Materials of Protein-Based Biologically Active Molecules. Biomedical Chemistry: Research and Methods, 2(3), e00096.


  1. Bigi, A., Boanini, E. (2017) Functionalized biomimetic calcium phosphates for bone tissue repair. Journal of Applied Biomaterials and Functional Materials; 15(4), 313-325. DOI
  2. Esposti, L.D., Carella, F., Adamiano, A., Tampieri, A., Iafisco, M. (2018) Calcium phosphate-based nanosystems for advanced targeted nanomedicine. Journal Drug Development and Industrial Pharmacy, 44(8),1223-1238. DOI
  3. Parent, M., Baradari, H., Champion, E., Damia, C., Viana-Trecant, M. (2017) Design of calcium phosphate ceramics for drug delivery applications in bone diseases: A review. Journal of Controlled Release, 28(252),1-17. DOI
  4. Bose, S., Tarafder, S. (2012) Calcium phosphate ceramic systems in growth factor and drug delivery for bone tissue engineering: A review. Acta Biomaterialia, 8, 1401-1421. DOI
  5. Bigi, A., Boanini, E. (2018) Calcium phosphates as delivery systems for bisphosphonates. Journal of Functional Biomaterials, 9(1), 6. DOI
  6. Li, W.M., Su, C.W., Chen, Y.W., Chen, S.Y. (2014) In situ DOX-calcium phosphate mineralized Q1 CPT-amphiphilic gelatin nanoparticle for intracellular controlled sequential release of multiple drugs. Acta Biomaterialia, 15, 191-199. DOI
  7. Chena, G., Liuc, B., Liua, H., Zhanga, H., Yanga, K., Wanga, Q., Dingb, J., Chang, F. (2018) Calcium phosphate cement loaded with 10% vancomycin delivering highearly and late local antibiotic сoncentration in vitro. Orthopaedics and traumatology: Surgery and research, 104, 1271-1275. DOI
  8. Uchida, K., Sugo, K., Nakajima, T., Nakawaki, M., Takano, S., Nagura, N., Takaso, M., Urabe, K. (2018) In Vivo release of vancomycin from calcium phosphate cement. BioMed Research International, 4560647. DOI
  9. Peter, B., Pioletti, D.P., Lairb, S., Bujoli, B., Pilet, P., Janvier, P., Guicheux, J., Zambelli, P.-Y, Bouler, J.-M., Gauthier, O. (2005) Calcium phosphate drug delivery system: influence of local zoledronate release on bone implant osteointegration. Bone, 36, 52-60. DOI
  10. Barroug, A., Kuhn, L.T., Gerstenfeld, L.C., Glimcher, M.J. (2004). Interactions of cisplatin with calcium phosphate nanoparticles: in vitro controlled adsorption and release. Journal of Orthopaedic Research, 22, 703-708. DOI
  11. Poli, E., Magnaudeix, A., Damia, C., Lalloué, F., Chaleix, V., Champion, E., Sol, V. (2019) Advanced protocol to functionalize CaP bioceramic surface with peptide sequences and effect on murine pre-osteoblast cells proliferation. Bioorganic and Medicinal Chemistry Letters, 29, 1069-1073. DOI
  12. Borcard, F., Staedler, D., Comas, H., Juillerat, F.K., Sturzenegger, P.N., Heuberger, R., Gonzenbach, U.T., Juillerat-Jeanneret, L., Gerber-Lemaire, S. (2012) Chemical functionalization of bioceramics to enhance endothelial cells adhesion for tissue engineering. Journal of Medicinal Chemistry, 27, 55(18):7988-97. DOI
  13. Dolci L.S., Panzavolta, S., Torricelli, P., Albertini, B., Sicuro, L., Fini, M., Bigi, A., Passerini, N. (2019) Modulation of Alendronate release from a calcium phosphate bone cement: An in vitro osteoblast-osteoclast co-culture study. International Journal of Pharmaceutics, 554, 245-255. DOI
  14. Stigter, M., Bezemera, J., Groot, K., Layrolle, P. (2004) Incorporation of different antibiotics into carbonated hydroxyapatite coatings on titanium implants, release and antibiotic efficacy. Journal of Controlled Release, 99, 127-137. DOI
  15. Gurin, A.N., Komlev, V.S., Fadeeva, I.V., Barinov, S.M. (2010) Octacalcium phosphate - precursor of biomineralization, novel bone scaffold. Stomatologiia, 4, 65-72.
  16. Komlev, V.S., Barinov, S.M., Bozo, I.I., Deev, R.V., Eremin, I.I., Fedotov, A.Y., Gurin, A.N., Khromova, N.V., Kopnin, P.B., Kuvshinova, E.A., Mamonov, V.E., Rybko, V.A., Sergeeva, N.S., Teterina, A.Y., Zorin, V.L. (2014) Bioceramics composed of octacalcium phosphate demonstrate enhanced biological behavior. ACS Applied Materials and Interfaces, 6, 16610−16620. DOI
  17. Dorozhkin, S.V. (2013) Calcium orthophosphate-based bioceramics. Materials, 6 (9), 3840-3942. DOI
  18. Navarro, M., Michiardi, A., Castano, O., Pianeil, J.A. (2008) Biomaterials in orthopaedies. Journal of the Royal Society Interface, 5, 1137-1158. DOI
  19. Lin, X., Groot, K., Wang, D., Hu, D., Wismeijer, D., Liu, Y. (2015) A review paper on biomimetic calcium phosphate coatings. The Open Biomedical Engineering Journal, 9, 56-64. DOI
  20. Su, Y., Luo, C., Zhang, Z., Hermawan, H., Zhu, D., Huang, J., Liang, Y., Li, G., Ren, L. (2018) Bioinspired surface functionalization of metallic biomaterials. Journal of the Mechanical Behavior of Biomedical Materials, 77, 90-105. DOI
  21. Lee, J.Y., Choi, B., Wu, B., Lee, M. (2013) Customized biomimetic scaffolds created by indirect three-dimensional printing for tissue engineering. Biofabrication, 5, 045003. DOI
  22. Habibovic, P., Van Der Valk, C.M., Van Blitterswijk, C.A., De Groot, K. (2004) Influence of octacalcium phosphate coating on osteoinductive properties of biomaterials. Journal of materials science: materials in medicine, 15, 373-380.
  23. Shin, K., Acri, T., Geary, S., Salem, A.K. (2017) Biomimetic mineralization of biomaterials using simulated body fluids for bone tissue engineering and regenerative medicine. Tissue engineering, 23(19-20), 1169-1180. DOI
  24. Mirhadi, B., Mehdikhani, B., Askari, N. (2011) Synthesis of nano-sized β-tricalcium phosphate via wet precipitation. Processing and Application of Ceramics, 5(4), 193-198. DOI
  25. Fadeeva, I.V., Gafurov, M.R., Filippov, Ya.Yu., Davydova, G.A., Savintseva, I.V., Fomin, A.S, Petrakova, N.V., Antonova, O.S., Ahmetov, L.I., Gabbasov, B.F., Izotov, V.V., Orlinsky, S.B., Barinov, S.M. (2016) Copper-substituted tricalcium phosphates. Doklаdy Chemistry, 471(2), 384-387. DOI
  26. Fadeeva, I.V., Fomin, A.S., Barinov, S.M., Petrakova, N.V. (2016) Rus Patent 2578435. A61L27/02, A61L27/10, A61F2/28. The method of obtaining porous ceramics from calcium phosphates for the treatment of bone defects № 2015112518A.
  27. Komlev, V.S., Fedotov, A.Y. (2016) Rus Patent 2596504. A61L27/12, A61K6/00, A61F2/28, C35/447. The method of obtaining ceramics composed on octacalcium phosphate. № 2015122276.
  28. Termine, J.D., Posner, A.S. (1966) Infra-red determination of the percentage of crystallinity in apatitic calcium phosphates. Nature, 211, 6268.
  29. Tarasovich, B.N. (2012) IR spectra of the main classes of organic compounds. Reference materials. Lomonosov Moscow State University.
  30. Yu X., Qu, H., Knecht, D., Wei, M. (2009). Incorporation of bovine serum albumin into biomimetic coatings on titanium with high loading efficacy and its release behavior. Journal of Materials Science: Materials in Medicine, 20 (1), 287-294. DOI
  31. Liu, Y., Hunziker, E.B., Layrolle, P., Bruijn, J.D., De Groot, K. (2004) Bone morphogenetic protein 2 incorporated into biomimetic coatings retains its biological activity. Tissue engineering, 10(1-2), 101-108.
  32. Liu, Y., De Groot, K., Hunziker, E.B. (2005) BMP-2 liberated from biomimetic implant coatings induces and sustains direct ossification in an ectopic rat model. Bone, 36, 745 - 757.
  33. Yu, X., Qu, H., Knecht, D.A., Wei, M. (2009) Incorporation of bovine serum albumin into biomimetic coatings on titanium with high loading efficacy and its release behavior. Journal of Materials Science: Materials in Medicine, 20, 287-294. DOI
  34. Forsgren, J., Brohede, U., Stromme, M., Engqvist, H.K. (2011) Co-loading of bisphosphonates and antibiotics to a biomimetic hydroxyapatite coating. Biotechnology Letters, 33, 1265-1268. DOI
  35. Eliaz, N.,Metoki, N. (2017). Calcium phosphate bioceramics: A review of their history, structure, properties, coating technologies and biomedical applications. Materials, 10(4), 334. DOI
  36. Ducheyne, P., Qiu, Q. (1999) The effect of surface reactivity on bone formation and bone cell function. Biomaterials, 20, 2287-2303.
  37. Veresov, A.G., Putlyaev, V.I., Tretyakov, Y.D. (2004). Chemistry of inorganic biomaterials based on calcium phosphate. Russian Chemical Journal, XLVIII(4), 52-64.
  38. Vasserman, I.M. (1980) Chemical precipitation from solutions. L.: Chemistry.
  39. Barrere, F., Blitterswijk, C.A., De Groot, K., Layrolle, P. (2002) Nucleation of biomimetic Ca-P coatings on Ti6Al4V from a SBF_5 solution: influence of magnesium. Biomaterials, 23, 2211-2220.
  40. Barrere, F., Blitterswijk, C.A., De Groot, K., Layrolle, P. (2002) Influence of ionic strength and carbonate on the Ca-P coating formation from SBF_5 solution. Biomaterials, 23, 1921 - 1930.