Ultrastructural and morphometric study of bone tissue regeneration dynamics using a combination of octacalcium phosphate with chitosan and ampicillin for osteoplasty
DOI:
https://doi.org/10.26641/1997-9665.2022.4.51-59Keywords:
mandible/lower jaw, dentoalveolar apparatus, bone tissue, bone regeneration, osteoplastic materials, сhitosan, morphometric study, scanning electron microscopy, rabbits.Abstract
The relevance of this study is to provide a comprehensive understanding of the dynamics of bone tissue remodeling after the creation of a defect and subsequent filling with an osteoplastic material containing chitosan, a cationic linear polysaccharide. Despite the established clinical use of osteoplastic materials and the unique properties of chitosan, its impact on bone regeneration in the maxillofacial region, its mechanisms and dynamics, remain incompletely understood and require further clarification and detailed investigation. Aim. The aim of this study is to determine the dynamics of histoarchitectural remodeling of bone-ceramic regenerate following the transplantation of octacalcium phosphate in combination with chitosan and ampicillin into an experimental defect in the rabbit mandible. Methods. Adult male rabbits aged 6-7 months and weighing 2.5-3 kg were used for the study. The control group consisted of animals with a bone defect that healed under a blood clot. The experimental group consisted of rabbits in which the bone defect was filled with an osteotropic material containing octacalcium phosphate, chitosan acetate, and ampicillin (OCР-N-Chitosan-Ampicillin). Post-traumatic bone tissue status within the defect area was monitored for 84 days. Ultrastructural changes were studied using scanning electron microscopy. To determine changes in the regenerate composition, three parameters were counted. The data was analyzed using the Student's t-test, and a difference at p < 0.05 was considered statistically significant. Results. The OCР-N-Chitosan-Ampicillin material induced significant regenerative changes in the surface of the lower jaw experimental bone defect that occurred after the injury and correlated with the dynamics of changes in the relative volume of bone tissue, osteoplastic material, and connective tissue in the regenerate. Morphometric analysis of the relative volume of the regenerative components of the experimental defect established a phased nature of the dynamics of the studied changes. The osteocyte lacuno-canalicular system formed following implantation of the material exhibited characteristics of a typical structure. No foci of incomplete osteogenesis were observed. Compared to the control group, in the area of the external bone lamellae after application of the OCР -N-Chitosan-Ampicillin material, the osteons of the regenerate, in terms of structure and geometry, corresponded to the normal bone of the mandibular ramus. Conclusion. In the experimental group of animals where the defect was filled with OCР-N-Chitosan-Ampicillin material, an active increase in the relative volume of bone tissue in the regenerate was observed. This increase followed a similar dynamic trend as the control group, but it was significantly more intense and advanced than the control.
References
- Xue N, Ding X, Huang R, Jiang R, Huang H, Pan X, et al. Bone tissue engineering in the treatment of bone defects. Pharmaceuticals. 2022;15:879. https://doi.org/10.3390/ph15070879
- Valtanen, R. S., Yang, Y. P., Gurtner, G. C., Maloney, W. J., & Lowenberg, D. W. Synthetic and Bone tissue engineering graft substitutes: What is the future?. Injury. 2021;52 Suppl 2:S72-S77. https://doi.org/10.1016/j.injury.2020.07.040
- Aguilar A, Zein N, Harmouch E, Hafdi B, Bornert F, Offner D, et al. Application of chitosan in bone and dental engineering. Molecules. 2019;24(16):3009. https://doi.org/10.3390/molecules24163009
- Dodero A, Scarfi S, Mirata S, Sionkowska A, Vicini S, Alloisio M, et al. Effect of crosslinking type on the physical-chemical properties and biocompatibility of chitosan-based electrospun membranes. Polymers (Basel). 2021;13(5):831. https://doi.org/10.3390/polym13050831
- Roffi A, Kon E, Perdisa F, Fini M, Di Martino A, Parrilli A, et al. A composite chitosan-reinforced scaffold fails to provide osteochondral regeneration. Int J Mol Sci. 2019;20(9):2227. https://doi.org/10.3390/ijms20092227
- Migliorini F, Cuozzo F, Torsiello E, Spiezia F, Oliva F, Maffulli N. Autologous bone grafting in trauma and orthopaedic surgery: an evidence-based narrative review. J Clin Med. 2021;10:4347. https://doi.org/10.3390/jcm10194347
- Gillman CE, Jayasuriya AC. FDA-approved bone grafts and bone graft substitute devices in bone regeneration. Mater Sci Eng C. 2021;130:112466. https://doi.org/10.1016/j.msec.2021.112466
- Hamada S, Mori Y, Shiwaku Y, Hamai R, Tsuchiya K, Baba K, et al. Octacalcium phosphate/gelatin composite (OCP/Gel) enhances bone repair in a critical-sized transcortical femoral defect rat model. Clin Orthop Relat Res. 2022;480:2043–55. https://doi.org/10.1097/CORR.0000000000002257
- Jeong C-H, Kim J, Kim HS, Lim S-Y, Han D, Huser AJ, et al. Acceleration of bone formation by octacalcium phosphate composite in a rat tibia critical-sized defect. J Orthop Transl. 2022;37:100–12. https://doi.org/10.1016/j.jot.2022.09.007
- Maliki S, Sharma G, Kumar A, Moral-Zamorano M, Moradi O, Baselga J, et al. Chitosan as a tool for sustainable development: A mini review. Polymers (Basel). 2022;14(7):1475. https://doi.org/10.3390/polym14071475
- Ke CL, Deng FS, Chuang CY, Lin CH. Antimicrobial actions and applications of chitosan. Polymers (Basel). 2021;13(6):904. https://doi.org/10.3390/polym13060904
- Shah FA, Ruscsák K, Palmquist A. 50 years of scanning electron microscopy of bone – a comprehensive overview of the important discoveries made and insights gained into bone material properties in health, disease, and taphonomy. Bone Res. 2019;7:15. https://doi.org/10.1038/s41413-019-0053-z
- Goldstein JI, Newbury DE, Michael JR, Ritchie NWM, Scott JHJ, Joy DC. Scanning electron microscopy and X-Ray microanalysis. New York, NY: Springer New York; 2017. https://doi.org/10.1007/978-1-4939-6676-9
- Aescht E, Büchl-Zimmermann S, Burmester A, Dänhardt-Pfeiffer S, Desel C, Hamers C, et al. Romeis mikroskopische technik. Heidelberg: Spektrum Akademischer Verlag; 2010. https://doi.org/10.1007/978-3-8274-2254-5
- European Convention for the protection of vertebrate animals used for experimental and other scientific purposes. Strasburg: Council of Europe. 1986;123:52.
- Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the Protection of Animals Used for Scientific Purposes. Off J Eur Union. 2010;53(L276):33–79.
- Paramita P, Ramachandran M, Narashiman S, et al. Sol-gel based synthesis and biological properties of zinc integrated nano bioglass ceramics for bone tissue regeneration. J Mater Sci Mater Med. 2021;32(1):5. https://doi.org/10.1007/s10856-020-06478-3
- Gómez-Barrena E, Padilla-Eguiluz N, Rosset P, Gebhard F, Hernigou P, Baldini N, et al. Early efficacy evaluation of mesenchymal stromal cells (MSC) combined to biomaterials to treat long bone non-unions. Injury. 2020;51(1):S63-S73. https://doi.org/10.1016/j.injury.2020.02.070
- Yan Y, Cheng B, Chen K, Cui W, Qi J, Li X, Deng L. Enhanced osteogenesis of bone marrow-derived mesenchymal stem cells by a functionalized silk fibroin hydrogel for bone defect repair. Adv Healthc Mater. 2019;8(3):e1801043. https://doi.org/10.1002/adhm.201801043
- Pandini FE, Kubo FMM, Plepis AMdG, Martins VdCA, da Cunha MR, Silva VR, et al. In vivo study of nasal bone reconstruction with collagen, elastin and chitosan membranes in abstainer and alcoholic rats. Polymers. 2022; 14(1):188. https://doi.org/10.3390/polym14010188
- Silva SK, Plepis AMG, Martins VDCA, Horn MM, Buchaim DV, Buchaim RL, et al. Suitability of chitosan scaffolds with carbon nanotubes for bone defects treated with photobiomodulation. Int J Mol Sci. 2022;23(12):6503. https://doi.org/10.3390/ijms23126503
- European Pharmacopoeia. 10th ed. Council of Europe, European Directorate for the Quality of Medicines Healthcare; Strasbourg, France: 2019.
- The United States Pharmacopeial Convention . The United States Pharmacopeia: The National Formulary. The United States Pharmacopeial Convention; Rockville, MD, USA: 2018.
Downloads
Published
How to Cite
Issue
Section
License

This work is licensed under a Creative Commons Attribution 4.0 International License.
The authors reserve the right to authorship of their work and transfer to the Journal the right to the first publication of this work under the terms of a license Creative commons Attribution 4.0 International (CC BY 4.0), which allows other people to freely distribute the published work with a mandatory reference to the authors of the original work and the first publication of the work in this journal.By submitting a manuscript to the editorial office of the Journal ‘Morphologia’ authors agree to transfer the rights to protect and use the manuscript (all supplemental materials, particularly protected objects such as photos, drawings, diagrams, tables, etc.), including the reproduction in the press and distribution via the Internet; translation of the manuscript into any language; export and import of journal copies with the Authors’ article in order to make it available for public. Authors convey the rights mentioned above to the editorial office without any temporal or territorial limitation all over the world.
The Authors guarantee that they have the exclusive rights to use the material transferred to editorial office. Editors are not responsible to third parties for contraventions of warranty given by the Authors. The considered rights are transferred to the editorial office since the moment when the current issue is signed for publishing. Reproduction of materials published in the Journal by other individuals and legal entities is possible only with the consent of Editorial office, with the obligatory indication of the full bibliographic reference of the primary publication. The Authors reserve the right to use the published material, its fragments and parts for teaching materials, oral presentations, dissertation thesis prepararion with obligatory bibliographic citation of the original paper. Electron copy of the published article, downloaded from official journal web-site in .pdf format may be put by authors on the official web-site of their institutions, any other official resources with open access.
