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Low-level laser therapy improves bone formation: stereology findings for osteoporosis in rat model

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Abstract

Low-level laser therapy (LLLT) benefits bone metabolism, but its use needs to be standardized. We evaluated the effects of LLLT on bone defects in calvaria of ovariectomized rats. Stereology was used to calculate tissue repair volume (V tr ), density of trabecular bone volume (Vv t ), total volume of newly formed trabecular bone (Vtot), and the area occupied by collagen fibers (A C ). Fifty-four Wistar rats were submitted to bilateral ovariectomy, and bone defects were created in calvaria after 150 days. The animals were divided into nine groups (n = 6), and 24 h after defects, the treatment started with a 780-nm low-intensity GaAlAs laser: G1, G2, and G3 received 3 sessions of 0, 20, and 30 J/cm2 respectively; G4, G5, and G6 received 6 sessions of 0, 20, and 30 J/cm2, respectively; and G7, G8, and G9 received 12 sessions of 0, 20, and 30 J/cm2, respectively. A normal distribution was found for all of the data. The test used to verify the normality was the Kolmogorov-Smirnov (KS, p > 0.05). The one-way ANOVA followed by Tukey’s post hoc test was used for data processing. A difference of p < 0.05 was considered statistically significant. Groups G2 and G1 showed significance for V tr , Vv t , Vtot, and (A C ). Results were significant for (Vv t ) and (Vtot) between G3 and G1. There were no significant results between G5 and G4 as well as between G8 and G7. Groups G6 and G4 results showed statistical difference for V tr , Vv t , Vtot, and (A C ). Groups G9 and G7 showed significance for V tr , Vv t , Vtot, and (A C ). In conclusion, there was new bone formation in the groups that received 20 and 30 J/cm2 when compared to control groups, but over time, the dose of 30 J/cm2 showed better stereological parameters when compared to 20 J/cm2.

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References

  1. NIH Consensus Development Panel on Osteoporosis Prevention, Diagnosis, and Therapy (2001) Osteoporosis prevention, diagnosis, and therapy. JAMA 285:785–795

    Article  Google Scholar 

  2. Payne JB, Reinhardt RA, Nummikoski PV, Patil KD (1999) Longitudinal alveolar bone loss in postmenopausal osteoporotic/osteopenic women. Osteoporos Int 10:34–40

    Article  CAS  PubMed  Google Scholar 

  3. Martínez-Maestre MÁ, González-Cejudo C, Machuca G, Torrejón R, Castelo-Branco C (2010) Periodontitis and osteoporosis: a systematic review. Climacteric 13:523–529

    Article  PubMed  Google Scholar 

  4. Dodd DZ, Rowe DJ (2013) The relationship between postmenopausal osteoporosis and periodontal disease. J Dent Hyg 87:336–344

    PubMed  Google Scholar 

  5. Pereira FM, Rodrigues VP, Oliveira AE, Brito LM, Lopes FF (2015) Association between periodontal changes and osteoporosis in postmenopausal women. Climacteric 18:311–315

    Article  CAS  PubMed  Google Scholar 

  6. von Wowern N, Klausen B, Kollerup G (1994) Osteoporosis: a risk factor in periodontal disease. J Periodontol 65:1134–1138

    Article  Google Scholar 

  7. Karu T (1989) Photobiology of low-power laser effects. Health Phys 56:691–704

    Article  CAS  PubMed  Google Scholar 

  8. Eduardo CP (2010) Lasers em odontologia. Guanabara Koogan, Rio de Janeiro

    Google Scholar 

  9. Pretel H, Lizarelli RF, Ramalho LT (2007) Effect of low-level laser therapy on bone repair: histological study in rats. Lasers Surg Med 39:788–796

    Article  PubMed  Google Scholar 

  10. Stein A, Benayahu D, Maltz L, Oron U (2005) Low-level laser irradiation promotes proliferation and differentiation of human osteoblasts in vitro. Photomed Laser Surg 23:161–166

    Article  CAS  PubMed  Google Scholar 

  11. Lirani-Galvão AP, Jorgetti V, Silva OL (2006) Comparative study of how low-level laser therapy and low-intensity pulsed ultrasound affect bone repair in rats. Photomed Laser Surg 24:735–740

    Article  PubMed  Google Scholar 

  12. Poppi RR, Silva AL, Nacer RS, Vieira RP, Oliveira LVF, Júnior NSF, Carvalho PTC (2011) Evaluation of the osteogenic effect of low-level laser therapy (808 nm and 660 nm) on bone defects induced in the femurs of female rats submitted to ovariectomy. Lasers Med Sci 26:515–522

    Article  Google Scholar 

  13. Freire MRS, Almeida D, Santos JN, Sarmento VA (2010) The photobiomodulation in the bone repair after radiotherapy: experimental study in rats. Proc. SPIE 7552, Mechanisms for Low-Light Therapy V, 75520H, doi:10.1117/12.842314

  14. Pinheiro ALB, Oliveira MG, Martins PPM, Ramalho LMP, Oliveira MAM, Novaes Júnior A, Nicolau RA (2001) Biomodulatory effects of LLLT on bone regeneration. Laser Ther 13:73–79

    Article  Google Scholar 

  15. Marcondes FK, Bianchi FJ, Tanno AP (2002) Determination of the estrous cycle phases of rats: some helpful considerations. Braz J Biol 62:609–614

    Article  CAS  PubMed  Google Scholar 

  16. Chicarelli M, Ramos FMM, Manzi FR, Novaes PD, Bóscolo FN, Almeida SM (2007) Effect of gamma rays on the bone repair process in rats with estrogen deficiency. Braz Oral Res 21:75–80

    Article  PubMed  Google Scholar 

  17. Fávaro-Pípi E, Ribeiro DA, Ribeiro JU, Bossini P, Oliveira P, Parizotto NA, Tim C, de Araújo HS, Renno AC (2011) Low-level laser therapy induces differential expression of osteogenic genes during bone repair in rats. Photomed Laser Surg 29:311–317

    Article  PubMed  Google Scholar 

  18. Nyengaard JR, Gundersen HJG (1992) The isector: a simple and direct method for generating isotropic, uniform random sections from small pieces. J Microsc 165:427–431

    Article  Google Scholar 

  19. Gundersen HJD (2002) The smooth fractionator. J Microsc 207:191–210

    Article  CAS  PubMed  Google Scholar 

  20. Lelovas PP, Xanthos TT, Thoma SE, Lyritis GP, Dontas IA (2008) The laboratory rat as an animal model for osteoporosis research. Comp Med 58:424–430

    CAS  PubMed Central  PubMed  Google Scholar 

  21. Kalu DN (1991) The ovariectomized rat model of postmenopausal bone loss. Bone Miner 15:171–192

    Article  Google Scholar 

  22. Frost HM, Jee WS (1992) On the rat model of human osteopenias and osteoporosis. Bone Miner 18:227–236

    Article  CAS  PubMed  Google Scholar 

  23. Wronski TJ, Dann LM, Horner SL (1989) Time course of vertebral osteopenia in ovariectomized rats. Bone 10:295–301

    Article  CAS  PubMed  Google Scholar 

  24. Wronski TJ, Dann LM, Scott K, Cintrón M (1989) Long-term effects of ovariectomy and aging on the rat skeleton. Calcif Tissue Int 45:360–366

    Article  CAS  PubMed  Google Scholar 

  25. Li M, Shen Y, Wronski TJ (1997) Time course of femoral neck osteopenia in ovariectomized rats. Bone 20:55–61

    Article  CAS  PubMed  Google Scholar 

  26. Schimitz JP, Hollinger JO (1986) The critical size defect as an experimental model for craniomandibulofacial nonunions. Clin Orthop Relat Res 205:299–308

    Google Scholar 

  27. Rawlinson SCF, Boyde A, Davis GR, Howell PGT, Hughes FJ, Kingsmill VJ (2009) Ovariectomy vs. hypofunction: their effects on rat mandibular bone. J Dent Res 88:615–620

    Article  CAS  PubMed  Google Scholar 

  28. Dai Q-G, Zhang P, Wu Y-Q, Ma X-H, Pang J, Jiang L-Y, Fang B (2014) Ovariectomy induces osteoporosis in the maxillary alveolar bone: an in vivo micro-CT and histomorphometric analysis in rats. Oral Dis 20:514–520

    Article  PubMed  Google Scholar 

  29. Silva AGP (2007) www.aulas.e-agps.info/estereologia/estereologia.pdf. Acessed 26 June 2014

  30. Howard CV, Reed MG (2010) Unbiased stereology. QTP Publications, Liverpool

    Google Scholar 

  31. Karu TI, Lubart R (2000) Effects of low-power light on biological systems. V. SPIE Proc 4159:1–17

    Google Scholar 

  32. Renno AC, Moura FM, Santos NS, Tirico RP, Bossini PS, Parizotto NA (2006) Effects of 830-nm laser light on preventing bone loss after ovariectomy. Photomed Laser Surg 24:642–645

    Article  PubMed  Google Scholar 

  33. Pires-Oliveira DA, Oliveira RF, Amadei SU, Pacheco-Soares C, Rocha RF (2010) Laser 904 nm action on bone repair in rats with osteoporosis. Osteoporos Int 21:2109–2114

    Article  CAS  PubMed  Google Scholar 

  34. Siessere S, Sousa LG, Issa JP, Iyomasa MM, Pitol DL, Barbosa AP, Semprini M, Sebald W, Bentley MV, Regalo SC (2011) Application of low-level laser irradiation (LLLI) and rhBMP-2 in critical bone defect of ovariectomized rats: histomorphometric evaluation. Photomed Laser Surg 29:453–458

    Article  CAS  PubMed  Google Scholar 

  35. Bossini PS, Renno ACM, Ribeiro DA, Fangel R, Ribeiro AC, Lahoz MA, Parizotto NA (2012) Low level laser therapy (830 nm) improves bone repair in osteoporotic rats: similar outcomes at two different dosages. Exp Gerontol 47:136–142

    Article  PubMed  Google Scholar 

  36. Rosa AP, de Sousa LG, Regalo SC, Issa JP, Barbosa AP, Pitol DL, de Oliveira RH, de Vasconcelos PB, Dias FJ, Chimello DT, Siéssere S (2012) Effects of the combination of low-level laser irradiation and recombinant human bone morphogenetic protein-2 in bone repair. Lasers Med Sci 27:971–977

    Article  PubMed  Google Scholar 

  37. Pinheiro ALB, Gerbi MEMM (2006) Photoengineering of bone repair process. Photomed Laser Surg 24:169–178

    Article  CAS  PubMed  Google Scholar 

  38. Tim CR, Pinto KN, Rossi BR, Fernandes K, Matsumoto MA, Parizotto NA, Rennó CN (2014) Low-level laser therapy enhances the expression of osteogenic factors during bone repairs in rats. Lasers Med Sci 29:146–156

    Article  Google Scholar 

  39. Sella VR, Bomfim FR, Machado PC, Silva Morsoleto MJ, Chohfi M, Plapler H (2015) Effect of low-level laser therapy on bone repair: a randomized controlled experimental study. Lasers Med Sci 30:1061–1068

    Article  PubMed  Google Scholar 

  40. Theodoro LH, Caiado RC, Longo M, Novaes VC, Zanini NA, Ervolino E, de Almeida JM, Garcia VG (2014) Effectiveness of the diode laser in the treatment of ligature-induced periodontitis in rats: a histopathological, histometric, and immunohistochemical study. Lasers Med Sci Apr 15

  41. Soares LG, Marques AM, Guarda MG, Aciole JM, dos Santos JN, Pinheiro AL (2014) Influence of the λ780nm laser light on the repair of surgical bone defects grafted or not with biphasic synthetic micro-granular hydroxylapatite + beta-calcium triphosphate. J Photochem Photobiol B 131:16–23

    Article  CAS  PubMed  Google Scholar 

  42. Fernandes KR, Ribeiro DA, Rodrigues NC, Tim C, Santos AA, Parizotto NA, Araujo HS, Driusso P, Rennó AC (2013) Effects of low-level laser therapy on the expression of osteogenic genes related in the initial stages of bone defects in rats. J Biomed Opt 18:038002. doi:10.1117/1

    Article  PubMed  Google Scholar 

  43. Almeida AL, Medeiros IL, Cunha MJ, Sbrana MC, Oliveira PG, Esper LA (2013) The effect of low-level laser on bone healing in critical size defects treated with or without autogenous bone graft: an experimental study in rat calvaria. Clin Oral Implants Res 25:1131–1136. doi:10.1111/clr.12239

    Article  PubMed  Google Scholar 

  44. Xu M, Deng T, Mo F, Deng B, Lam W, Deng P, Zhang X, Liu S (2009) Low-intensity pulsed laser irradiation affects RANKL and OPG mRNA expression in rat calvarial cells. Photomed Laser Surg 27:309–315

    Article  CAS  PubMed  Google Scholar 

  45. Saad A, Yamany ME, Abbas O, Yehia M (2010) Possible role of low level laser therapy on bone turnover in ovariectomized rats. Endocr Regul 44:155–163

    Article  CAS  PubMed  Google Scholar 

  46. Kanenari M, Zhao J, Abiko Y (2011) Enhancement of microtubule-associated protein-1 alpha gene expression in osteoblasts by low level laser irradiation. Laser Ther 20:47–51

    Article  PubMed Central  PubMed  Google Scholar 

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Acknowledgments

We thank FAPESP (São Paulo Research Foundation—Processes No. 2011/50686-0 and 2012/10184-9), CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico), and Pró-Reitoria de Pesquisa da Universidade de São Paulo for financial support.

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The authors declare that they have no conflict of interest.

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“All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.”

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Correspondence to Selma Siessere.

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Scalize, P.H., de Sousa, L.G., Regalo, S.C.H. et al. Low-level laser therapy improves bone formation: stereology findings for osteoporosis in rat model. Lasers Med Sci 30, 1599–1607 (2015). https://doi.org/10.1007/s10103-015-1773-y

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  • DOI: https://doi.org/10.1007/s10103-015-1773-y

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