Porous nanocomposite scaffolds prepared from gelatin and hydroxyapatite for bone tissue engineering: direct mixing and biomimetic

Document Type : Original Article

Authors
1 amirkabir university
2 university Amirkabir
3 AMIRKABIR UNIVERSITY
4 Amirkabir University
Abstract
In this study, gelatin-hydroxyapatite (GEL/HA) composites as biological organic-inorganic composites were made up of the capability for use in hard tissue to be examined by two methods, direct mixing and biomimetic. In the direct mixing method, after the synthesis of hydroxyapatite, the resulting powder was mixed with gelatin; in the biomimetic method, hydroxyapatite was synthesized in the presence of gelatin. The thin layer composite substrates were prepared with a thickness of 2 mm from the resulting mixture by combining methods including casting solvent and freeze-drying. The three-dimensional scaffolds were modified by glutaraldehyde as a crosslinked factor. The results showed that scaffolds have a high porosity of approximately 88% and are interconnected with holes. According to the SEM image, the average pore size is around 100 μm. A study of Infrared spectroscopy (FT-IR) and X-ray diffraction (XRD) indicated that the apatite phase formation non-stoichiometric low crystalline along with extensive replacement of carbonate ions on the network that is very biologically close to apatite phase and two composite components not only physically but also chemically interact with each other. Compressive strength test results also show that both scaffolds have mechanical properties similar to the cancellous bone. Young's modulus and density increase, and porosity and water absorption decrease by increasing the content of the hydroxyapatite composite. Despite the suitability of both methods, it seems that the biomimetic-made scaffolds are more suitable due to the higher density, higher tolerance levels of stress and Young's modulus, lower crystallinity, and replacement of carbonate ions appropriate method.
Keywords
Subjects

[1] J.L. González-Carrasco, “Metals as bone repair
materialsˮ, In Bone Repair Biomaterials, Woodhead,
2009, pp. 154-193.
[2] C.H. C hang, C.Y. Lin, C.H. Chang, F.H. Liu,
Y.T. Huang, and Y.S. Liao, “Enhanced biomedical
applicability of ZrO2–SiO2 ceramic composites in 3D
printed bone scaffoldsˮ, Sci. Rep., vol. 12, no. 1, pp.
1-11, 2022.
[3] S. Samavedi, A.R. Whittington, and A.S. Goldstein,
“Calcium phosphate ceramics in bone tissue
engineering: a review of properties and their influence
on cell behaviorˮ, Acta Biomater., vol. 9, no. 9, pp.
8037-8045, 2013.
[4] G.R. Rodríguez, T.M.F. Patrício, and J.D. López,
“Natural polymers for bone repairˮ, In Bone Repair
Biomaterials, Woodhead, 2019, pp. 199-232.
[5] B. Amiri, M. Ghollasi, M. Shahrousvand, M.
Kamali, and A. Salimi, “Osteoblast differentiation
of mesenchymal stem cells on modified PES-PEG
electrospun fibrous composites loaded with Zn2SiO4
bioceramic nanoparticlesˮ, Differentiation, vol. 92, no.
4, pp. 148-158, 2016.
[6] P. Bhattacharjee, B. Kundu, D. Naskar, T.K. Maiti,
D. Bhattacharya, and S.C. Kundu, “Nanofibrous
nonmulberry silk/PVA scaffold for osteoinduction and
osseointegrationˮ, Biopolymers, vol. 103, no. 5, pp.
271-284, 2015.
[7] R. Zeinali, L.J. Del Valle, J. Torras, and J. Puiggalí,
“Recent progress on biodegradable tissue engineering
scaffolds prepared by thermally-induced phase
separation [8] M.L. Chinta, A. Velidandi, N.P.P. Pabbathi, S.
Dahariya, and S.R. Parcha, “Assessment of properties,
applications and limitations of scaffolds based on
cellulose and its derivatives for cartilage tissue
engineering: A reviewˮ, Int. J. Biol. Macromol., vol.
175, pp. 495-515, 2021.
[9] R. Logith Kumar, A. Keshav Narayan, S. Dhivya,
A. Chawla, S. Saravanan, and N. Selvamurugan, “A
review of chitosan and its derivatives in bone tissue
engineeringˮ, Carbohyd. Polym., vol. 151, pp. 172-
188, 2016.
[10] G.A.N. Atia, H.K. Shalaby, M. Zehravi, M.M.
Ghobashy, H.A.N. Attia, Z. Ahmad et al., “Drug-loaded
chitosan scaffolds for periodontal tissue regenerationˮ,
Polymers, vol. 14, no. 15, pp. 3192, 2022.
[11] A. Yıldız, A.A. Kara, and F. Acartürk, “Peptide-protein
based nanofibers in pharmaceutical and biomedical
applicationsˮ, Int. J. Biol. Macromol., vol. 148, pp.
1084-1097, 2020.
[12] T.R. de Lima Nascimento, M.M. de Amoedo Campos
Velo, C.F. Silva, S. Costa Cruz, B.L.C. Gondim, R.F.L.
Mondelli, and L.R.C. Castellano, “Current applications
of biopolymer-based scaffolds and nanofibers as drug
delivery systemsˮ, Curr. Pharm. Design, vol. 25, no.
37, pp. 3997-4012, 2019.
[13] A.E. Purushothaman, K. Thakur, and B.
Kandasubramanian, “Development of highly porous,
Electrostatic force assisted nanofiber fabrication for
biological applicationsˮ, Int. J. Polym. Mater. Polym.
Biomater., vol. 69, no. 8, pp. 477-504, 2020.
[14] H. Syed and A.K. Sailaja, “Biodegradable polymers
and their applications in drug delivery systemˮ,
Interactions, vol. 3, no. 4, 2018.
[15] M. Fernández and J. Orozco, “Advances in
functionalized photosensitive polymeric nanocarriersˮ,
Polymers, vol. 13, no. 15, pp. 2464, 2021.
[16] S. Gundu, N. Varshney, A.K. Sahi, and S.K. Mahto,
“Recent developments of biomaterial scaffolds and
regenerative approaches for craniomaxillofacial bone
tissue engineeringˮ, J. Polym. Res., vol. 29, no. 3, pp.
1-23, 2022.
[17] S. Kanwar and S. Vijayavenkataraman, “Design of
3D printed scaffolds for bone tissue engineering: A
reviewˮ, Bioprinting, vol. 24, pp. e00167, 2021.
[18] Y. Yamaguchi, T. Matsuno, A. Miyazawa, Y.
Hashimoto, and T. Satomi, “Bioactivity evaluation of
biphasic hydroxyapatite bone substitutes immersed
and grown with supersaturated calcium phosphate
solutionˮ, Materials, vol. 14, no. 18, pp. 5143, 2021.
[19] R.A. Youness, M.S. Amer, and M.A. Taha, “Tribo-
mechanical measurements and in vivo performance
of zirconia-containing biphasic calcium phosphate
material implanted in a rat model for bone replacement
applicationsˮ, Mater. Chem. Phys., vol. 285, pp.
126085, 2022.
[20] N.N.F.N.M. Noordin, N. Ahmad, M. Mariatti, B.H.
Yahaya, A.R. Sulaiman, and Z.A.A. Hamid, “A review
on bioceramics scaffolds for bone defect in different
types of animal models: HA and β-TCPˮ, Biomed.
Phys. Eng. Express., vol. 8, no. 5, 2022.
[21] M. Azami, F. Moztarzadeh, and M. Tahriri,
“Preparation, characterization and mechanical
properties of controlled porous gelatin/hydroxyapatite
nanocomposite through layer solvent casting combined
with freeze-drying and lamination techniquesˮ, J.
Porous Mater., vol. 17. no. 3, pp. 313-320, 2010.
[22] J.E. Devin, M.A. Attawia, and C.T. Laurencin, “Three-
dimensional degradable porous polymer-ceramic
matrices for use in bone repairˮ, J. Biomater. Sci.
Polym. Ed., vol. 7, no. 8, pp. 661-669, 1996.
[23] J.M. Taboas, R.D. Maddox, P.H. Krebsbach, and S.J.
Hollister, “Indirect solid free form fabrication of local
and global porous, biomimetic and composite 3D
polymer-ceramic scaffoldsˮ, Biomaterials, vol. 24, no.
1, pp. 181-194, 2003.
[24] K. Grassie and Y. Khan, “Bone tissue engineeringˮ, In
Musculoskeletal Tissue Engineering, Elsevier, 2022,
pp. 1-40.
[25] B. Johari, M. Ahmadzadehzarajabad, M. Azami, M.
Kazemi, M. Soleimani, S. Kargozar et al., “Repair of
rat critical size calvarial defect using osteoblast-like
and umbilical vein endothelial cells seeded in gelatin/
hydroxyapatite scaffoldsˮ, J. Biomed Mater. Res. Part
A, vol. 104, no. 7, pp. 1770-1778, 2016.
[26] D. Jeyachandran, L. Li, R. Fairag, L. Haglund, and M.
Cerruti, “Simple fabrication and enhanced bioactivity
of bioglass-poly(lactic-co-glycolic acid) composite
scaffolds with matrix microporosityˮ, Macromol.
Mater. Eng., vol. 307, no. 7, pp. 2100863, 2022.
[27] G. Cao, Nanostructures and Nanomaterials: Synthesis,
Properties and Applications, Imperial college, 2004.
[28] M.C. Chang, C.C. Ko, and W.H. Douglas, “Preparation
of hydroxyapatite-gelatin nanocompositeˮ,
Biomaterials, vol. 24, no. 17, pp. 2853-2862, 2003.
[29] E. Landi, A. Tampieri, G. Celotti, R. Langenati, M.
Sandri, and S. Sprio, “Nucleation of biomimetic apatite
in synthetic body fluids: dense and porous scaffold
developmentˮ, Biomaterials, vol. 26, no. 16, pp. 2835-
2845, 2005.
[30] C. Shu, Y. Xianzhu, X. Zhangyin, X. Guohua, L.
Hong, and Y. Kangde, “Synthesis and sintering of
nanocrystalline hydroxyapatite powders by gelatin-based precipitation methodˮ, Ceram. Int., vol. 33, no.
2, pp. 193-196, 2007.
[31] W.B. Liu, S.X. Qu, R. Shen, C.X. Jiang, X.H. Li, B.
Feng et al., “Influence of pH values on preparation of
hydroxyapatite/gelatin compositesˮ, J. Mater. Sci., vol.
41, no. 6, pp. 1851-1853, 2006.
[32] J.E. Won, A. El-Fiqi, S.H. Jegal, C.M. Han, E.J. Lee,
J.C. Knowles et al., “Gelatin-apatite bone mimetic co-
precipitates incorporated within biopolymer matrix to
improve mechanical and biological properties useful
for hard tissue repairˮ, J. Biomater. Appl., vol. 28, no.
8, pp. 1213-1225, 2014.
[33] R. Hodgskinson and J.D. Currey, “The effect of
variation in structure on the Young’s modulus of
cancellous bone: a comparison of human and non-
human materialˮ, Proc. Inst. Mech. Eng. Part H: J.
Eng. Med., vol. 204, no. 2, pp. 115-121, 1990.