Biomedical Chemistry: Research and Methods 2019, 2(3), e00096

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

E.A. Kuvshinova1*, N.V. Petrakova2, N.S. Sergeeva1, V.A. Kirsanova1, I.K. Sviridova1, A.Y. Teterina2, V.S. Komlev2, A.D. Kaprin1

1Herzen Moscow Oncology Research Institute – branch of the National Medical Research Radiological Centre,
3 the 2-nd Botkinskiy proezd, Moscow, 125284 Russia; *e-mail: beliay@mail.ru

2Baikov Institute of Metallurgy and Material Science, Russian Academy of Science,
49 Leninskiy prospect 49, Moscow, 119334 Russia

Keywords:functionalization; biomimetic coating; biologically active molecules; octacalcium phosphate; β-tricalcium phosphate

DOI:10.18097/BMCRM00096

The whole version of this paper is available in Russian.

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.
Figure 1. SEM-photographs of the experimental materials: (a, b) – OCP, (a - х200, b - х2000),
(c, d) – β-TCP (c - х200, d – х2500).
Figure 2. Micrographs of the β-TCP ceramic surface: (a) – initial granules (х32000),
(b) – granules soaked in 2xSBF solution during 24 h (х40000).
Figure 3. FTIR spectra of the initial TCP granules (1) and soaked in 2xSBF during 24 h (2),
(a) – an overall view, (b) – detailing of the phosphate band.
Figure 4. Comparison of BSA loading efficacy on the OCP, β-TCPmod. and β-TCP surface with protein concentration in SCS and DPBS 3.0 mg/ml.
Figure 5. FTIR spectra of materials before and after BSA functionalisation in various solutions:
(a) - β-TCP, 1 - β-TCP initial, 2 - β-TCP in SCS, 3 - β-TCPmod. in SCS, 4 - β-TCP in DPBS,
5 - β-TCPmod. in DPBS; (с) - OCP, 1- OCP initial, 2 - OCP in SCS, 3 - OCP in DPBS,
(b) and (d) – detailing of FTIR-bands at 1350-1850 cm-1.

CLOSE
Table 1. The ionic composition of used buffer solutions.

CLOSE
Table 2. Comparison of BSA amount loaded and released (for 6 days) from 1 mg of OCP.

CLOSE
Table 3. Characteristics of used incorporation solutions: SCS, SBFmod., DPBS, TRIS.

FUNDING

The reported study was funded by RFBR within to the research project № 18-29-11052.

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