Histopathological and Micro-CT Scan Evaluation of the Repair of Large Bone Lesions using Apatite Carbonate and Titanium-Containing Bioactive Glass in a Rabbit Model


Published: Apr 18, 2025
Keywords:
Bone regeneration carbonate apatite bioactive glass titanium histopathology rabbit
S Moradi
A Tabatabaei Naeini
M Sayyari
A Shaikhzadeh
Abstract

In recent years, several techniques have been used to reconstruct large bone fractures, including various scaffolds and bone reparative materials, but the results have not been remarkable. Therefore, this study investigates and compares the effect of new scaffolds based on apatite carbonate and bioactive glasses containing titanium in promoting the healing process of large bone lesions in animal models in laboratory conditions. After constructing the scaffolds, in vivo studies were performed by creating four circular defects in the calvarial bones of 10 adult New Zealand rabbits, and then the cavities were randomly filled with bioactive glass powders containing titanium and apatite carbonate. The survival and proliferation of mesenchymal stem cells through the MTT method and bone defects in the studied groups were evaluated using different diagnostic imaging methods and histological analyses. The XRD and FTIR results confirmed the high purity of the fabricated Bg-Ca and Bg-Ti scaffolds. In the MTT method, the scaffolds made at a concentration of 10 mg/ml had no cytotoxicity against MSCs. Also, in total, microCT scan analyzes and histological findings showed a significant improvement in the healing process in rabbits treated with Bg-Ti and Bg-Ca compared to the group that received the Bg scaffold alone and in the control group.

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References
Abulyazied, D. E., A. M. Alturki, R. A. Youness and H. Abomostafa
(2021). “Synthesis, structural and biomedical characterization of hydroxyapatite/
borosilicate bioactive glass nanocomposites.” Journal
of Inorganic and Organometallic Polymers and Materials 31: 4077-
Al-Bakhsh, B. A. J., F. Shafiei, A. Hashemian, K. Shekofteh, B. Bolhari
and M. Behroozibakhsh (2019). “In-vitro bioactivity evaluation and
physical properties of an epoxy-based dental sealer reinforced with
synthesized fluorine-substituted hydroxyapatite, hydroxyapatite and
bioactive glass nanofillers.” Bioactive Materials 4: 322-333.
Ansari, M. (2019). “Bone tissue regeneration: biology, strategies and interface
studies.” Progress in biomaterials 8(4): 223-237.
Bal, Z., T. Kaito, F. Korkusuz and H. Yoshikawa (2020). “Bone regeneration
with hydroxyapatite-based biomaterials.” Emergent Materials
: 521-544.
Battafarano, G., M. Rossi, V. De Martino, F. Marampon, L. Borro, A. Secinaro
and A. Del Fattore (2021). “Strategies for bone regeneration:
from graft to tissue engineering.” International journal of molecular
sciences 22(3): 1128.
Bellucci, D., V. Cannillo, A. Anesi, R. Salvatori, L. Chiarini, T. Manfredini
and D. Zaffe (2018). “Bone regeneration by novel bioactive glasses
containing strontium and/or magnesium: a preliminary in-vivo
study.” Materials 11(11): 2223.
Bellucci, D., A. Sola and V. Cannillo (2013). “Bioactive glass/ZrO2 composites
for orthopaedic applications.” Biomedical Materials 9(1):
Bigham-Sadegh, A., H. S. Torkestani, S. Sharifi and S. Shirian (2020).
“Effects of concurrent use of royal jelly with hydroxyapatite on bone
healing in rabbit model: radiological and histopathological evaluation.”
Heliyon 6(7): e04547.
Carlini, A. S., L. Adamiak and N. C. Gianneschi (2016). “Biosynthetic
polymers as functional materials.” Macromolecules 49(12): 4379-
Currey, J. D. (2013). Bones. Bones, Princeton University Press.
de Melo Pereira, D. and P. Habibovic (2018). “Biomineralization‐Inspired
Material Design for Bone Regeneration.” Advanced Healthcare Materials
(22): 1800700.
Ege, D., K. Zheng and A. R. Boccaccini (2022). “Borate Bioactive Glasses
(BBG): Bone Regeneration, Wound Healing Applications, and Future
Directions.” ACS Applied Bio Materials 5(8): 3608-3622.
El-Rashidy, A. A., J. A. Roether, L. Harhaus, U. Kneser and A. R. Boccaccini
(2017). “Regenerating bone with bioactive glass scaffolds: A
review of in vivo studies in bone defect models.” Acta biomaterialia
: 1-28.
Eldeeb, A. E., S. Salah, M. Mabrouk, M. S. Amer and N. A. Elkasabgy
(2022). “Dual-drug delivery via zein in situ forming implants augmented
with titanium-doped bioactive glass for bone regeneration:
preparation, in vitro characterization, and in vivo evaluation.” Pharmaceutics
(2): 274.
Fernandes, H. R., A. Gaddam, A. Rebelo, D. Brazete, G. E. Stan and J. M.
Ferreira (2018). “Bioactive glasses and glass-ceramics for healthcare
applications in bone regeneration and tissue engineering.” Materials
(12): 2530.
Hu, K. and B. R. Olsen (2016). “The roles of vascular endothelial growth
factor in bone repair and regeneration.” Bone 91: 30-38.
Jalili, M., A. T. Naeini, M. S. A. Khafi, K. Mys and A. Khoshzaban (2022).
“Effectiveness of Reinforced 45S5 Bioglass with Yttria-Stabilized
Tetragonal Zirconia and Iron in the Regeneration of Rabbit Calvarial
Defects.” Acta Veterinaria Eurasia 48(1): 35-48.
Jiang, S., M. Wang and J. He (2021). “A review of biomimetic scaffolds
for bone regeneration: Toward a cell‐free strategy.” Bioengineering &
Translational Medicine 6(2): e10206.
Kashirina, A., Y. Yao, Y. Liu and J. Leng (2019). “Biopolymers as bone
substitutes: A review.” Biomaterials science 7(10): 3961-3983.
Kopecká, I. and E. Svobodová (2014). “Methodology for infrared spectroscopy
analysis of sandwich multilayer samples of historical materials.”
Heritage Science 2(1): 1-8.
Kouroupis, D., A. Kyrkou, E. Triantafyllidi, M. Katsimpoulas, G. Chalepakis,
A. Goussia, A. Georgoulis, C. Murphy and T. Fotsis (2016).
“Generation of stem cell-based bioartificial anterior cruciate ligament
(ACL) grafts for effective ACL rupture repair.” Stem cell research
(2): 448-457.
Krishnakumar, G. S., A. Roffi, D. Reale, E. Kon and G. Filardo (2017).
“Clinical application of bone morphogenetic proteins for bone healing:
a systematic review.” International orthopaedics 41: 1073-1083.
Lett, J. A., S. Sagadevan, I. Fatimah, M. E. Hoque, Y. Lokanathan, E.
Léonard, S. F. Alshahateet, R. Schirhagl and W. C. Oh (2021). “Recent
advances in natural polymer-based hydroxyapatite scaffolds:
Properties and applications.” European Polymer Journal 148: 110360.
Lü, S., X. Bai, H. Liu, P. Ning, Z. Wang, C. Gao, B. Ni and M. Liu (2017).
“An injectable and self-healing hydrogel with covalent cross-linking
in vivo for cranial bone repair.” Journal of Materials Chemistry B
(20): 3739-3748.
Mirjalili, F., S. A. Manafi and R. Rashadi (2020). “Assessment of cytotoxicity
and mechanical properties of bioactive 45S5 fluoroapatite-glass
nanocomposite.” Journal of Ceramic Science and Engineering 8(4):
-101.
Mistry, S., D. Kundu, S. Datta and D. Basu (2011). “Comparison of bioactive
glass coated and hydroxyapatite coated titanium dental implants
in the human jaw bone.” Australian dental journal 56(1): 68-75.
Oryan, A., S. Monazzah and A. Bigham-Sadegh (2015). “Bone injury and
fracture healing biology.” Biomedical and environmental sciences
(1): 57-71.
Radulescu, D.-E., I. A. Neacsu, A.-M. Grumezescu and E. Andronescu
(2022). “Novel trends into the development of natural hydroxyapatite-
based polymeric composites for bone tissue engineering.” Polymers 14(5): 899.
Ramesh, N., S. C. Moratti and G. J. Dias (2018). “Hydroxyapatite-polymer
biocomposites for bone regeneration: A review of current
trends.” Journal of Biomedical Materials Research Part B: Applied
Biomaterials 106(5): 2046-2057.
Sari, M., Chotimah, I. D. Ana and Y. Yusuf (2022). “Cell viability assay
and surface morphology analysis of carbonated hydroxyapatite/
honeycomb/titanium alloy coatings for bone implant applications.”
Bioengineering 9(7): 325.
Schneible, J. D. (2020). A Material Toolbox for Advanced Therapeutics,
North Carolina State University.
Shamsi, M., A. Salimi, M. Ghallasi and R. Halabian (2018). “Effect of
synthetic biologically activated 45S5 glass nanoparticles on osteogenesis
differentiation of mesenchymal human bone marrow.” The
Scientific Journal of Iranian Blood Transfusion Organization 15(4):
-286.
Smith, S., O. ElKashty, F. Tamimi, S. D. Tran and M. Cerruti (2021).
“Titanium-Containing Silicate-Based Sol-Gel Bioactive Glass: Development,
Characterization, and Applications.” Langmuir 37(49):
-14253.
van Gestel, N. A., J. Geurts, D. J. Hulsen, B. van Rietbergen, S. Hofmann
and J. Arts (2015). “Clinical applications of S53P4 bioactive glass
in bone healing and osteomyelitic treatment: a literature review.”
BioMed research international 2015.
Yadegari, M., S. A. Bigham, S. Farhang and S. Farhangi (2020). “Effect
of Nano-hydroxyapatite Concurrent with Bone Marrow Treatment on
Experimental Bone Defect Healing in Rabbit: Radiological and Histopathological
Evaluation.”
Yun, J.-H., J.-H. Yoo, S.-H. Choi, M.-H. Lee, S.-J. Lee, S. U. Song and
N.-S. Oh (2012). “Synergistic effect of bone marrow-derived mesenchymal
stem cells and platelet-rich plasma on bone regeneration
of calvarial defects in rabbits.” Tissue Engineering and Regenerative
Medicine 9(1): 17-23.
Zeng, J.-H., S.-W. Liu, L. Xiong, P. Qiu, L.-H. Ding, S.-L. Xiong, J.-T. Li,
X.-G. Liao and Z.-M. Tang (2018). “Scaffolds for the repair of bone
defects in clinical studies: A systematic review.” Journal of orthopaedic
surgery and research 13(1): 1-14.
Zhu, T., Y. Cui, M. Zhang, D. Zhao, G. Liu and J. Ding (2020). “Engineered
three-dimensional scaffolds for enhanced bone regeneration in
osteonecrosis.” Bioactive materials 5(3): 584-601.