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Membrane-free bone regeneration using biphasic calcium phosphate, heterologous fibrin biopolymer, and photobiomodulation: an in vivo study

Bruna Trazzi Pagani1, Filiphe Gustavo Daré1, Beatriz de Oliveira Fernandes1, Karina Oliveira Santos2, Benedito Barraviera3,4,5, Rui Seabra Ferreira Jr3,4,5, Murilo Priori Alcalde6, Marco Antonio Hungaro Duarte6, Daniela Vieira Buchaim3,4,7,8,9, Rogerio Leone Buchaim2,8,*  [ + show more ]

J Venom Anim Toxins incl Trop Dis, 2026, 32:e20260012
Received: 05 Februay 2026 | Accepted: 01 July 2026 | Published: 03 August 2026
https://doi.org/10.1590/1678-9199-JVATITD-2026-0012

Abstract

Background: Although bone tissue possesses inherent regenerative capacity, criticalsized defects require grafts for complete functional repair.This in vivo study evaluated the bone repair process using laser photobiomodulation therapy (PBM) in defects filled with a combination of hydroxyapatite, β-tricalcium phosphate and heterologous fibrin biopolymer (HFB). Methods: Thirty male rats were divided into three groups: biomaterial alone (BG), biomaterial + HFB (BBG), and biomaterial + HFB + PBM (BBPG). A 5-mm circular calvarial osteotomy was performed and filled according to each protocol. In BBPG, an 830- nm laser was applied immediately post-surgery and three times weekly until euthanasia at 14 or 42 days. Analyses included micro-CT, histomorphology, histomorphometry, and polarized light microscopy of collagen fibers. Results: Micro-CT showed centripetal bone regeneration restricted to defect margins, with biomaterial particles persisting centrally. Histologically, new bone progressed from immature trabecular architecture at day 14 to a mature lamellar conformation by day 42, notably in BBPG. All groups showed a significant temporal increase in new bone percentage. BBPG demonstrated superior bone growth at 42 days (26.64 ± 2.15%) compared to BG (14.85 ± 1.63%) and BBG (20.05 ± 1.70%). The birefringence of the collagen fibers showed a color transition from red to yellowish-green during the analyzed periods. Conclusion: The combination of the biomaterial, fibrin biopolymer and photobiomodulation significantly enhanced bone defect repair and matrix maturation without barrier membranes, presenting high translational potential for cost-effective clinical applications in regenerative medicine.

 

Keywords: Biocompatible materials, Bone regeneration, Photobiomodulation, Fibrin sealant, Biopolymers

 

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