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Imaging of nano-hydroxyapatite/chitosan scaffolds using a cone beam computed tomography device on rat calvarial defects with histological verification

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Abstract

Objectives

Τhis study aims at determining the ability of cone beam computed tomography (CBCT) to visualize critical-size defects (CSD) created at rat calvaria and filled with 75/25 w/w nano-hydroxyapatite/chitosan (nHAp/CS) scaffolds, prior to their histological investigation.

Materials and methods

Thirty adult Sprague Dawley rats, 15 males and 15 females, were used. Two CSD, 5 mm in diameter, were bilaterally trephined in the parietal bone. The right CSD was filled with nHAp/CS scaffold, while the left CSD remained empty, as the control group. Two female rats died post-operatively. Rats were euthanized at 2, 4, and 8 weeks post-surgery. Twenty-eight specimens (15 × 2 × 10 mm) were resected—containing both CSDs—and then scanned using a NewTom VGi CBCT imaging unit (Verona, Italy). The manufacturer’s software trace region profile tool (NNT v6.2, Verona, Italy) was used in selected axial slices. The greyscale value (in VGiHU) and the traced/selected region of interest (ROI, in mm2) of those areas were automatically calculated. Subsequently, all specimens were histologically examined.

Results

An increased VGiHU (P = 0.000), was observed in the experimental group relative to the control group. The ROI of CSD (in mm2) was significantly reduced (P = 0.001) from the fourth to the eighth week in both groups. No statistically significant difference between male and female rats (P = 0.188) was observed with respect to VGiHU.

Conclusions

The nHAp/CS scaffolds are easily visualized using a particular high-resolution CBCT device.

Clinical relevance

Both the CBCT measurements and also the histological results suggest that the nHAp/CS scaffold presence contributes to new bone formation in rat calvarial CSD.

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Acknowledgments

Hyperbranched poly (ethyleneimine) was kindly donated by BASF Hellas S.A.

Funding

This research is co-financed by Greece and the European Union (European Social Fund—ESF) through the Operational Programme “Human Resources Development, Education and Lifelong Learning” in the context of the project “Strengthening Human Resources Research Potential via Doctorate Research” (MIS-5000432), implemented by the State Scholarships Foundation (ΙΚΥ).

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Authors and Affiliations

  1. Department of Oral Diagnosis and Radiology, Faculty of Dentistry, National and Kapodistrian University of Athens, 2 Thivon Str, 11527, Goudi, Athens, Greece

    Emmanouil Chatzipetros, Zafeiroula Yfanti, Catherine Donta, Spyros Damaskos & Kostas Tsiklakis

  2. Department Oral and Maxillofacial Surgery, Faculty of Dentistry, National and Kapodistrian University of Athens, Athens, Greece

    Panos Christopoulos

  3. Department of Immunochistochemistry and Molecular Biology, 401 Military Hospital of Athens, Athens, Greece

    Evangelos Tsiambas

  4. Institute of Nanoscience and Nanotechnology, National Centre for Scientific Research “Demokritos”, Aghia Paraskevi, Attiki, Greece

    Dimitris Tsiourvas

  5. Department of Oral Medicine and Pathology, Faculty of Dentistry, National and Kapodistrian University of Athens, Athens, Greece

    Eleni-Marina Kalogirou & Konstantinos I. Tosios

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  1. Emmanouil Chatzipetros
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  2. Zafeiroula Yfanti
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Correspondence to Emmanouil Chatzipetros.

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Ethical approval

All animal experimental protocols and procedures were approved by the Directorate of Agricultural and Veterinary Policy with protocol number 1181/2-03-2017 and registration code EL 25 BIO 05, Athens, Greece.

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Chatzipetros, E., Yfanti, Z., Christopoulos, P. et al. Imaging of nano-hydroxyapatite/chitosan scaffolds using a cone beam computed tomography device on rat calvarial defects with histological verification. Clin Oral Invest 24, 437–446 (2020). https://doi.org/10.1007/s00784-019-02939-4

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