Effects of calcium phosphates incorporation on structural, thermal and drug-delivery properties of collagen:chitosan scaffolds

Mirella Romanelli Vicente Bertolo, Virginia Conceição Amaro Martins, Ana Maria de Guzzi Plepis

Abstract


In this study, we evaluated how different procedures of calcium phosphate synthesis and its incorporation in collagen:chitosan scaffolds could affect their structural and thermal properties, aiming the obtention of homogeneous scaffolds which can act as drug delivery vehicles in bone tissue engineering. Therefore, three different scaffold preparation procedures were developed, changing the order of addition of the components: in CC-CNPM1 and CC-CNPM2, calcium phosphate synthesis was performed in situ in the chitosan gel (1%, w/w) followed by mixture with collagen (1%, w/w), with changes in the reagents used for calcium phosphate formation; in CC-CNPM3 procedure, calcium phosphate was synthesized ex situ and then incorporated into the collagen gel, in which chitosan in powder was mixed. In all procedures, 5% (in dry mass) of ciprofloxacin was incorporated. FTIR analysis confirmed the presence of calcium phosphate in all scaffolds. DSC curves showed that collagen denaturation temperature (Td) increased with calcium incorporation. SEM photomicrographs of scaffolds cross-section revealed porous scaffolds with calcium phosphate grains internally distributed in the polymeric matrix. XRD diffractograms indicated that the calcium phosphates obtained are hydroxyapatite. The pore size distribution was more homogeneous for CC-CNPM3, which also stood out for its smaller porosity and lower absorption in PBS. These results indicate that the in situ or ex situ phosphate incorporation in the scaffolds had a great influence on its structural properties, which also had consequences for ciprofloxacin release. CC-CNPM3 released a smaller amount of antibiotic (30%), but its release profile was better described by all the tested models.


Keywords


Calcium phosphate, collagen, chitosan, bone regeneration, ciprofloxacin release

References


Lanza R, Langer R, Vacanti J, Principles of Tissue Engineering. Academic Press, New York, NY, USA, 3rd edition (2007).

Wojnar R, Bone and Cartilage – its Structure and Physical Properties. Biomechanics of Hard Tissues: Modeling, Testing, and Materials, ed. by Andreas Ochsner e Waqar Ahmed, WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim, Germany (2010).

Velasco MA, Narváez-Tovar CA, Garzón-Alvarado DA, Design, Materials, and Mechanobiology of Biodegradable Scaffolds for Bone Tissue Engineering. BioMed Res Int, 2015 :729076, (2015).

Fisher MB, Mauck RL, Tissue Engineering and Regenerative Medicine: Recent Innovations and the Transition to Translation. Tissue Eng Part B Rev, 19 :1-13 (2013).

Zugravu MV et al., Physical properties and in vitro evaluation of collagen–chitosan–calcium phosphate microparticle-based scaffolds for bone tissue regeneration. J Biomed Appl, 28(4) :566-579 (2012).

Nwe N, Furuike T, Tamura H, The mechanical and biological properties of chitosan scaffolds for tissue regeneration templates are significantly enhanced by chitosan from Gongronella butleri. Materials, 2 :374–398 (2009).

Wu S et al., Biomimetic porous scaffolds for bone tissue engineering. Mat Sci Eng R, 80 :1–36 (2014).

Rodrígues-Vázquez M et al., Chitosan and Its Potential Use as a Scaffold for Tissue Engineering in Regenerative Medicine. Biomed Res Int, 821279 (2015).

Dong C & Yonggang Lv, Application of Collagen Scaffold in Tissue Engineering: Recent Advances and New Perspectives. Polymers, 8(2) :42 (2016).

Gopi D, Kanimozhi K, Kavitha L, Opuntia ficus indica peel derived pectin mediated hydroxyapatite nanoparticles: Synthesis, spectral characterization, biological and antimicrobial activities. Spectrochim Acta, Part A, 141 :135-143 (2015).

Chen DZ et al., Dynamic mechanical properties and in vitro bioactivity of PHBHV/HA nanocomposite. Compos Sci Technol, 67 :1617–1626 (2007).

Pelin IM et al., Preparation and characterization of a hydroxyapatite-collagen composite as component for injectable bone substitute. Mater Sci Eng C, 29 :2188–2194 (2009).

Elhendawi H et al., Effect of synthesis temperature on the crystallization and growth of in situ prepared nanohydroxyapatite in chitosan matrix. ISRN Biomaterials, 2014 :897468 (2014).

Ghomi H, Fathi MH, Edris H, Preparation of nanostructure hydroxyapatite scaffold for tissue engineering applications. J. Sol-Gel Sci Technol, 58 :642-650 (2011).

Keeney M et al., The ability of a collagen/calcium phosphate scaffold to act as its own vector for gene delivery and to promote bone formation via transfection with VEGF165. Biomaterials, 31 :2893-2902 (2010).

Inzana JA et al., 3D printing of composite calcium phosphate and collagen scaffolds for bone regeneration. Biomaterials, 35 :4026-4034 (2014).

Ferreira AM, Gentile P, Chiono V, Ciardelli G, Collagen for bone tissue regeneration. Acta Biomater, 8 :3191–3200 (2012).

Nandi SK et al., Local antibiotic delivery systems for the treatment of osteomyelitis: a review. Mater Sci Eng C, 29 :2478-2485 (2009).

Zhang X et al., Teicoplanin-loaded borate bioactive glass implants for treating chronic bone infection in a rabbit tibia osteomyelitis model. Biomaterials, 31 :5865-5874 (2010).

Horn MM, Martins VCA, Plepis AMG, Interaction of anionic collagen with chitosan: Effect on thermal and morphological characteristics. Carbohydr Polym, 77 :239–243 (2009).

Kurita K, Chitin and chitosan: Functional biopolymers from marine crustaceans. Mar Biotechnol, 8(3) :203-226 (2006).

Lavertu M et al., A validated 1H NMR method for the determination of the degree of deacetylation of chitosan. J Pharm Biomed Anal, 32(6) :1149-1158 (2003).

Rinaudo M. Chitin and chitosan: Properties and applications. Prog Polym Sci, 31 :603-632 (2006).

Barrère F, Layrolle P, Van Blitterswijk CA, De Groot K, Biomimetic Calcium Phosphate Coatings on Ti6Al4V: A Crystal Growth Study of Octacalcium Phosphate and Inhibition by Mg2+ and HCO3-. Bone, 25(2) :107S-111S (1999).

Pereda M et al., Chitosan-gelatin composites and bi-layer films with potential antimicrobial activity. Food Hydrocoll, 25(5) :1372-1381 (2011).

Batista TM, Martins VCA, Plepis AMG, Thermal behavior of in vitro mineralized anionic collagen matrices. J Therm Anal Calorim, 95 :945-949 (2009).

Berzina-Cimdina L & Borodajenko N, Research of Calcium Phosphates Using Fourier Transform Infrared Spectroscopy, Infrared Spectroscopy - Materials Science, Engineering and Technology, Prof. Theophanides Theophile (Ed.), ISBN: 978-953-51-0537-4, InTech, 2012.

Kanapathipillai M et al., Synthesis and Characterization of Ionic Block Copolymer Templated Calcium Phosphate Nanocomposites. Chem Mater, 20 :5922-5932 (2008).

Koutsopoulos S. Synthesis and characterization of hydroxyapatite grains: A review study on the analytical methods. J Biomed Mater Res, 62(4) :600-612 (2002).

Li B, Huang Y, Wang Y, Zhou Y. Mineralization of Bone-like Apatite in Chitosan Hydrogel. Key Eng Mat, 434-435 :605-608 (2010).

Zhao H, Ma L, Gao C, Shen J. Fabrication and properties of mineralized collagen-chitosan/hydroxyapatite scaffolds. Polym Adv Technol, 19 :1590-1596 (2008).

Karageorgiou V & Kaplan D, Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials, 26(27) :5474–5491 (2005).

Liu LS, Thompson AY, Heidaran MA, Poser JW, Spiro RC. An osteoconductive collagen/hyaluronate matrix for bone regeneration. Biomaterials, 20(12) :1097–1108 (1999).

Chesnutt BM et al., Design and characterization of a novel chitosan/nanocrystalline calcium phosphate composite scaffold for bone regeneration. [Evaluation Studies]. J Biomed Mater Res Part A, 88(2) :491–502 (2008).

Martins VCA & Goissis G, Nonstoichiometric hydroxyapatite-anionic collagen composite as support for the double sustained release of gentamicin and norfloxacin/ciprofloxacin. Artif Organs, 24(3) :224-230 (2000).

Wallis SC et al., Interaction of norfloxacin with divalent and trivalent pharmaceutical cations: in vitro complexation and in vivo pharmacokinetic studies in the dog. J Pharm Sci, 85 :803-809 (1996).

Kelly DJ et al. Serum concentrations of penicillin, doxycycline, and ciprofloxacin during prolonged therapy in rhesus monkeys. J Infect Dis; 166 :1184-1187 (1992).

Paarakh MP et al., Release kinetics – concepts and applications. Int J Pharm Res Technol, 8 :12-20 (2018).

Shaikh HK, Kshirsagar RV & Patil SG. Mathematical models for drug release characterization: a review. Int J Pharm Pharm Sci, 4(4): 324-338 (2015).




DOI: https://doi.org/10.25061/2595-3931/IJAMB/2019.v2i2.32

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