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Raman study of the repair of surgical bone defects grafted with biphasic synthetic microgranular HA + β-calcium triphosphate and irradiated or not with λ780 nm laser

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Abstract

The treatment of bone loss due to different etiologic factors is difficult, and many techniques aim to improve repair, including a wide range of biomaterials and, recently, photobioengineering. This work aimed to assess, through Raman spectroscopy, the level of bone mineralization using the intensities of the Raman peaks of both inorganic (∼960, ∼1,070, and ∼1,077 cm−1) and organic (∼1,454 and ∼1,666 cm−1) contents of bone tissue. Forty rats were divided into four groups each subdivided into two subgroups according to the time of killing (15 and 30 days). Surgical bone defects were made on femur of each animal with a trephine drill. On animals of group Clot, the defect was filled only by blood clot; on group Laser, the defect filled with the clot was further irradiated. On animals of groups Biomaterial and Laser + Biomaterial, the defect was filled by biomaterial and the last one was further irradiated (λ780 nm, 70 mW, Φ ∼ 0.4 cm2, 20 J/cm2 session, 140 J/cm2 treatment) in four points around the defect at 48-h intervals and repeated for 2 weeks. At both 15th and 30th day following killing, samples were taken and analyzed by Raman spectroscopy. At the end of the experimental time, the intensities of both inorganic and organic contents were higher on group Laser + Biomaterial. It is concluded that the use of laser phototherapy associated to biomaterial was effective in improving bone healing on bone defects as a result of the increasing deposition of calcium hydroxyapatite measured by Raman spectroscopy.

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Notes

  1. Protocol 08.2010.

  2. About 2 months old, average weight 295 ± 25 g.

  3. Labina®, Purina, São Paulo, Brazil.

  4. INSIGHT Equipamentos Ltda—Monte Alegre, Ribeirão Preto, São Paulo, Brazil.

  5. 0.04 ml/100 g of atropine subcutaneously.

  6. 10 % ketamine (0.1 mL/100 g—Cetamin®, Syntec, São Paulo, Brazil) + 2 % xylazin (0.1 mL/100 g; Xilazin®, Syntec, São Paulo, Brazil).

  7. SIN, São Paulo, Brazil.

  8. NSK, Tochigi, Japan.

  9. Driller 600®, SIN, São Paulo, SP, Brazil.

  10. Pentabiotico®, 0.2 ml; Fort Dodge Animal Health, Overland Park, KS, USA.

  11. TwinFlex Evolution®, MMOptics, São Carlos, São Paulo, Brazil; λ780 nm, 70 mW, Φ ∼ 0.4 cm2, 20 J/cm2.

  12. Power Meter Thorlabs PM30-121, Thorlabs GmbH, Munich, Germany.

  13. Insight Equipamentos, model EB 248, Ribeirão Preto, SP, Brazil.

  14. SIN-DRILLER 600 BML, São Paulo, SP, Brazil.

  15. Andor Technology, model Shamrock SR-303i®, Belfast, Northern Ireland.

  16. B&W TEK, model BRM-785-0.30-100-0.22.s, Newark, DE, USA.

  17. B&W TEK, model BAC-100-785, Newark, DE, USA.

  18. Andor Technology, model IDUs® DU401A-BR-DD, Belfast, Northern Ireland.

  19. Andor Technology, Solis (i) software, Belfast, Northern Ireland.

  20. Intensity correction and wavenumber calibration.

  21. Oriel Instruments, model 63358, Strattford, CT, USA.

  22. The Mathworks, Newark, NJ, USA.

  23. Minitab, Belo Horizonte, MG, Brazil.

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Authors and Affiliations

  1. Center of Biophotonics, School of Dentistry, Federal University of Bahia, Av. Araújo Pinho, 62, Canela, Salvador, BA, 40110-150, Brazil

    Luiz Guilherme P. Soares, Aparecida Maria C. Marques, Artur Felipe S. Barbosa, Nicole R. Santos, Jouber Mateus S. Aciole, Caroline Mathias C. Souza & Antonio Luiz B. Pinheiro

  2. National Institute of Optics and Photonics, Physics Institute of São Carlos, University of São Paulo, São Carlos, SP, Brazil, 13560-970

    Aparecida Maria C. Marques & Antonio Luiz B. Pinheiro

  3. Camilo Castelo Branco University, Núcleo do Parque Tecnológico de São José dos Campos: Rod. Presidente Dutra Km 139, São José dos Campos, Eugênio de Melo, SP, 12247-004, Brazil

    Antonio Luiz B. Pinheiro & Landulfo Silveira Jr.

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  1. Luiz Guilherme P. Soares
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  2. Aparecida Maria C. Marques
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  3. Artur Felipe S. Barbosa
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  4. Nicole R. Santos
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  5. Jouber Mateus S. Aciole
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  6. Caroline Mathias C. Souza
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  7. Antonio Luiz B. Pinheiro
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  8. Landulfo Silveira Jr.
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Correspondence to Antonio Luiz B. Pinheiro.

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Soares, L.G.P., Marques, A.M.C., Barbosa, A.F.S. et al. Raman study of the repair of surgical bone defects grafted with biphasic synthetic microgranular HA + β-calcium triphosphate and irradiated or not with λ780 nm laser. Lasers Med Sci 29, 1539–1550 (2014). https://doi.org/10.1007/s10103-013-1297-2

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