Effects of Hydroxyapatite Pretreated with Two Sodium Fluoride Concentrations on Early Bone Healing in Beagle Dogs
7 affiliations
- 1Department of Histology and Embryology, Nihon University Graduate School of Dentistry at Matsudo, Matsudo 271-8587, Japan
- 2Department of Physiology, Saveetha Medical College and Hospital, Saveetha Institute of Medical and Technical Sciences, Chennai 600077, India
- 3Research Institute of Oral Science, Nihon University School of Dentistry at Matsudo, Matsudo 271-8587, Japan
- 4Department of Oral Pathology, Faculty of Dentistry, Jamia Millia Islamia, New Delhi 110025, India
- 5Japan Oral Medicine Organization, Tokyo 135-0033, Japan
- 6Department of Histology, Nihon University School of Dentistry at Matsudo, Matsudo 271-8587, Japan
- 7School of Dentistry, Kanagawa Dental University, Yokosuka 238-8580, Japan; [email protected]
Abstract
Hydroxyapatite (HA) is widely used as an osteoconductive bone substitute, and fluoride can influence bone-forming cells in a concentration-dependent manner. This study examined how pretreating resorbable HA with low or high concentrations of sodium fluoride (NaF) affects early mandibular bone healing in beagle dogs. Twelve beagle dogs were used in the project, including three in a pilot study and nine in the main experiment. At each observation time point (Days 4, 7, and 14), one dog was assigned to each of the three HA-based treatment conditions (HA, HA + 0.2% NaF, or HA + 4% NaF). Four experimental defects in each dog received the same assigned treatment, while four contralateral defects were left unfilled as controls. The four defects were considered within-animal subsamples rather than independent biological replicates, and all treatment comparisons were descriptive and exploratory. Before implantation, HA was exposed to the designated NaF solution for 3 min and then washed with 100 mL of distilled water. Healing was evaluated by H&E, Azan, Masson’s trichrome and toluidine blue staining, immunohistochemistry, and quantitative real-time RT-PCR. Histological examination showed progressive tissue and matrix organization around the implanted material, with more evident new bone and osteoblast-associated changes in the low-NaF group at Days 7 and 14. Runx2 immunoreactivity was more prominent in the early healing phase, and osteocalcin was more prominent in the later healing phase. Osteocalcin mRNA gradually increased until Day 14, whereas Runx2 mRNA was maximally expressed at Day 7. Both responses were strongest with low-NaF-pretreated HA. High-NaF-pretreated HA retained osteogenic activity but did not produce a response of the same magnitude. These exploratory findings suggest that NaF pretreatment concentration may influence early HA-associated bone healing, warranting confirmation in studies with independent animal-level replication.
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BibTeX
@article{suzuki2026effects,
title = {{Effects of Hydroxyapatite Pretreated with Two Sodium Fluoride Concentrations on Early Bone Healing in Beagle Dogs}},
author = {Yoshino Suzuki and Ujjal K. Bhawal and Aman Chowdhry and Takao Watanabe and Hiroyuki Okada and Kazunari Kimoto},
journal = {International Journal of Molecular Sciences},
year = {2026},
month = oct,
doi = {10.3390/ijms27219414},
url = {https://www.mdpi.com/1422-0067/27/21/9414},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Effects of Hydroxyapatite Pretreated with Two Sodium Fluoride Concentrations on Early Bone Healing in Beagle Dogs
AU - Yoshino Suzuki
AU - Ujjal K. Bhawal
AU - Aman Chowdhry
AU - Takao Watanabe
AU - Hiroyuki Okada
AU - Kazunari Kimoto
PY - 2026
DA - 2026/10/22
JO - International Journal of Molecular Sciences
DO - 10.3390/ijms27219414
UR - https://www.mdpi.com/1422-0067/27/21/9414
AB - Hydroxyapatite (HA) is widely used as an osteoconductive bone substitute, and fluoride can influence bone-forming cells in a concentration-dependent manner. This study examined how pretreating resorbable HA with low or high concentrations of sodium fluoride (NaF) affects early mandibular bone healing in beagle dogs. Twelve beagle dogs were used in the project, including three in a pilot study and nine in the main experiment. At each observation time point (Days 4, 7, and 14), one dog was assigned to each of the three HA-based treatment conditions (HA, HA + 0.2% NaF, or HA + 4% NaF). Four experimental defects in each dog received the same assigned treatment, while four contralateral defects were left unfilled as controls. The four defects were considered within-animal subsamples rather than independent biological replicates, and all treatment comparisons were descriptive and exploratory. Before implantation, HA was exposed to the designated NaF solution for 3 min and then washed with 100 mL of distilled water. Healing was evaluated by H&E, Azan, Masson’s trichrome and toluidine blue staining, immunohistochemistry, and quantitative real-time RT-PCR. Histological examination showed progressive tissue and matrix organization around the implanted material, with more evident new bone and osteoblast-associated changes in the low-NaF group at Days 7 and 14. Runx2 immunoreactivity was more prominent in the early healing phase, and osteocalcin was more prominent in the later healing phase. Osteocalcin mRNA gradually increased until Day 14, whereas Runx2 mRNA was maximally expressed at Day 7. Both responses were strongest with low-NaF-pretreated HA. High-NaF-pretreated HA retained osteogenic activity but did not produce a response of the same magnitude. These exploratory findings suggest that NaF pretreatment concentration may influence early HA-associated bone healing, warranting confirmation in studies with independent animal-level replication.
SN - 1422-0067
ER -