Single local injection of 3S-HMGB1 enhances early bone healing and titanium implant osseointegration in type 2 diabetic mice

Authors

DOI:

https://doi.org/10.1590/

Keywords:

Diabetes Mellitus, Oral Osseointegration, 3S-HMGB1, CXCR4

Abstract

Diabetes significantly impairs bone healing via several mechanisms, including sustained inflammation, oxidative stress, and poor osteogenic responses. Fully reduced isoform of High Mobility Group Box 1 with three serine substitutions (3S-HMGB1) has shown regenerative properties while being resistant to oxidative degradation. Objective:  This study evaluated the therapeutic potential of a single local injection of 3S-HMGB1 in early osseointegration under diabetic conditions in mice. Methodology:  A total of 48 male 129/Sv mice (24 non-diabetic [ND], 24 diabetic [D]) received titanium implants following maxillary first molar extraction. ND and D mice (n:12) were injected with either saline (control) or 3S-HMGB1 (0.75 mg/kg) into the fresh extraction socket. Osseointegration was evaluated at 7 and 21 days post-implantation using microCT, histology (bone-to-implant contact [BIC] and birefringence), and immunohistochemistry for Runx2 and CXCR4. Results:  ND controls exhibited early osteogenic activity, with a predominance of Runx2-positive cells at 7 days and successful osseointegration by 21 days. In contrast, D controls showed reduced numbers of Runx2-positive cells and markedly lower BV/TV, indicating compromised bone healing at 21 days. Treatment with 3S-HMGB1 resulted in significantly increased bone volume at the implant site in D animals (55.6±7.20% vs. 44.6±6.23%) and restored BIC from 44.9±9.32% (D controls) to 61.78±11.31% (D 3S-HMGB1), near ND levels (65.16±7.64%). Both ND and D groups treated with 3S-HMGB1 presented enhanced collagen organization. No significant differences were found in CXCR4 between groups; however, in D animals, a distinct peri-implant staining pattern suggested impaired recruitment despite preserved stem cell niches in the bone marrow. Conclusions:  Collectively, our findings indicate that a single injection of redox-stable 3S-HMGB1 may represent a promising regenerative strategy to mitigate early implant failure in diabetes. Future studies should explore sustained delivery approaches to enhance long-term outcomes.

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Published

2025-10-20

Issue

Vol. 33 (2025)

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Original Articles

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Copyright (c) 2025 Bhuvana Lakkasetter Chandrashekar, Alexandra Arteaga, Evelin Rios, Jimena Mora, Gustavo Pompermaier Garlet, Danieli C. Rodrigues, Claudia Cristina Biguetti

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How to Cite

Chandrashekar, B. L., Arteaga, A., Rios, E., Mora, J., Garlet, G. P., Rodrigues, D. C., & Biguetti, C. C. (2025). Single local injection of 3S-HMGB1 enhances early bone healing and titanium implant osseointegration in type 2 diabetic mice. Journal of Applied Oral Science, 33, e20250358. https://doi.org/10.1590/