We have used Raman analysis to assess bone healing on different models. Benefits on the isolated or combined use of mineral trioxide aggregate, bone morphogenetic proteins, guided bone regeneration and laser on bone repair have been reported, but not their combination. We studied peaks of hydroxyapatite and CH groups on defects grafted with MTA, treated or not with laser, BMPs, and GBR. Ninety rats were divided in 10 groups each, subdivided into three subgroups. Laser (λ850 nm) was applied at every other day for 2 weeks. Raman readings were taken at the surface of the defect. Statistical analysis (CHA) showed significant differences between all groups (p = 0.001) and between Group II and all other (p < 0.001), but not with Group X (p = 0.09). At day 21 differences were seen between all groups (p = 0.031) and between Groups VIII and X when compared with Groups VI (p = 0.03), V (p < 0.001), IV (p < 0.001), and IX (p = 0.04). At the end of the experimental period no significant differences were seen. On regards CH, significant differences were seen at the 15(th) day (p = 0.002) and between Group II and all other groups (p < 0.0001) but not with control. Advanced maturation on irradiated bone is because of increased secretion of calcium hydroxyapatite (CHA) that is indicative of greater calcification and resistance of the bone. We conclude that the association of the MTA with laser phototherapy (LPT) and/or not with GBR resulted in a better bone repair. The use of the MTA associated to IR LPT resulted in a more advanced and quality bone repair.
Fractures have different etiology and treatment and may be associated or not to bone losses. Laser light has been shown to improve bone healing. We aimed to assess, through Raman spectroscopy, the level of CHA (approximately 958 cm(-1)) on complete fractures animals treated with IRF treated or not with Low Level Laser Therapy-LLLT and associated or not to BMPs and GBR. Complete tibial fractures were created on 15 animals that were divided into five groups. LLLT (Laser Unit, Kondortech, São Carlos, SP, Brazil, lambda790 nm, 4 J/cm(2)/point, 40 mW, phi approximately 0.5 cm(2), 16 J/cm(2) session) started immediately after surgery and repeated at 48 h interval (2 weeks). Animal death occurred after 30 days. Raman spectroscopy was performed at the surface of the fracture. Our results showed significant differences between the groups IRF + BL /IRF_NBL (p = 0.05); between all experimental groups and untreated bone; bone/IRF + BL; IRF + BL + Bio + GBR; IRF + BL + LLLT; IRF + BL + Bio + GBR + LLLT; IRF_NBL (p < 0.001, all); IRF_NBL/IRF + BL + LLLT (p = 0.03); IRF_NBL/IRF + BL + Bio + GBR + LLLT (p = 0.02); IRF + BL/IRF + BL + LLLT (p = 0.04); IRF + BL/IRF + BL + Bio + GBR + LLLT (p = 0.002); IRF + BL + Bio + GBR/IRF + BL + Bio + GBR + LLLT (p = 0.05). It is concluded that the use of NIR LLLT associated to BMPs and GBR was effective in improving bone healing on fractured bones due to increased levels of CHA.
Hepatitis C has become one of the higher health problems around the world. Near-infrared Raman spectroscopy had been used to spectrally differentiate among health human blood serum from the one with hepatitis C contaminationin vitro. In this study a Raman spectrometer with 80 mW, 830 nm excitation, liquid-nitrogen cooled CCD and imaging spectrograph were used to collect Raman scattering from 24 blood samples (14 healthy and 10 diseased) with collection time of 120 s. It has been used an algorithm based on the Principal Components Analysis (PCA) for main spectral features identification and Mahalanobis distance for blood spectrum classification depending on the serology. It was observed that the highest spectral differences between the two types of human blood serum were found in 1002, 1169, 1262 and 1348 cm−1Raman bands. The spectral analysis using multivariate statistics presented good results when compared to classical diagnosis for viral hepatitis C, showing that Raman spectroscopy can classify human blood serum spectrum in one of the two categories by identifying biochemical alterations that occur in the presence of viral infections.
The aim of the present study was to assess, by Raman spectroscopy and laser fluorescence, the repair of surgical fractures fixed with wire osteosynthesis treated or not with infrared laser (λ780 nm, 50 mW, 4 × 4 J/cm(2) =16 J/cm(2), ϕ=0.5 cm(2), CW) associated or not to the use of hydroxyapatite and guided bone regeneration. Surgical tibial fractures were created under general anesthesia on 15 rabbits that were divided into five groups, maintained on individual cages, at day/night cycle, fed with solid laboratory pelted diet, and had water ad libitum. The fractures in groups II, III, IV, and V were fixed with wires. Animals in groups III and V were grafted with hydroxyapatite (HA) and guided bone regeneration (GBR) technique used. Animals in groups IV and V were irradiated at every other day during 2 weeks (4 × 4 J/cm(2), 16 J/cm(2) =112 J/cm(2)). Observation time was that of 30 days. After animal death, specimens were taken and kept in liquid nitrogen and used for Raman spectroscopy. The Raman results showed basal readings of 1,234.38 ± 220. Groups WO+B+L showed higher readings (1,680.22 ± 822) and group WO+B the lowest (501.425 ± 328). Fluorescence data showed basal readings of 5.83333 ± 0.7. Groups WO showed higher readings (6.91667 ± 0.9) and group WO+B+L the lowest (1.66667 ± 0.5). There were significant differences between groups on both cases (p<0.05). Pearson correlation was negative and significant (R (2) = -0.60; p<0.001), and it was indicative that, when the Raman peaks of calcium hydroxyapatite (CHA) are increased, the level of fluorescence is reduced. It is concluded that the use of near-infrared lasertherapy associated to HA graft and GBR was effective in improving bone healing on fractured bones as a result of the increasing deposition of CHA measured by Raman spectroscopy and decrease of the organic components as shown by the fluorescence readings.
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