Marine collagen scaffolds and photobiomodulation on bone healing process in a model of calvaria defects.

INTRODUCTION: Collagen from marine esponges has been used as a promising material for tissue engineering proposals. Similarly, photobiomodulation (PBM) is able of modulating inflammatory processes after an injury, accelerating soft and hard tissue healing and stimulating neoangiogenesis. However, the effects of the associated treatments on bone tissue healing have not been studied yet. In this context, the present study aimed to evaluate the biological temporal modifications (using two experimental periods) of marine sponge collagen or sponging (SPG) based scaffold and PBM on newly formed bone using a calvaria bone defect model. MATERIAL AND METHODS: Wistar rats were distributed into two groups: SPG or SPG/PBM and euthanized into two different experimental periods (15 and 45 days post-surgery). A cranial critical bone defect was used to evaluate the effects of the treatments. Histology, histomorfometry and immunohistological analysis were performed. RESULTS: Histological findings demonstrated that SPG/PBM-treated animals, 45 days post-surgery, demonstrated a higher amount of connective and newly formed bone tissue at the region of the defect compared to CG. Notwithstanding, no difference among groups were observed in the histomorphometry. Interestingly, for both anti-transforming growth factor-beta (TGF-ß) and anti-vascular endothelial growth factor (VEGF) immunostaining, higher values for SPG/PBM, at 45 days post-surgery could be observed. CONCLUSION: It can be concluded that the associated treatment can be considered as a promising therapeutical intervention.

Influence of the incorporation of marine spongin into a Biosilicate®: an in vitro study.

The combination of different biomaterials can be a promising intervention for the composites manufacture, mainly by adding functional and structural characteristics of each material and guarantee the advantages of the use of these composites. In this context, the aim of this study was to develop and evaluated the influence of the incorporation of marine spongin (SPG) into Biosilicate® (BS) in different proportions be used during bone repair. For this purpose, it was to develop and investigate different BS/SPG formulations for physico-chemical and morphological characteristics by pH, loss mass, Fourier transform infrared spectrometer (FTIR) and scanning electron microscope (SEM) analysis. Additionally, the influence of these composites on cell viability, proliferation, and alkaline phosphatase (ALP) activity were investigated. The results revealed that the pH values of all BS groups (with or without SPG) increased over time. A significant mass loss was observed in all composites, mainly with higher SPG percentages. Additionaly, SEM micrographies demonstrated fibers of SPG into BS and material degradation over time. Moreover, FTIR spectral analysis revealed characteristic peaks of PMMA, BS, and SPG in BS/SPG composites. BS/SPG groups demonstrated a positive effect for fibroblast proliferation after 3 and 7 days of culture. Additionally, BS and BS/SPG formulations (at 10% and 20% of SPG) presented similar values of osteoblasts viability and proliferation after 7 days of culture. Furthermore, ALP activity demonstrated no significant difference between BS and BS/SPG scaffolds, at any composition. Based on the present in vitro results, it can be concluded that the incorporation of SPG into BS was possible and produced an improvement in the physical-chemical characteristics and in the biological performance of the graft especially the formulation with 80/20 and 90/10. Future research should focus on in vivo evaluations of this novel composite.

Incorporation of Collagen from Marine Sponges (Spongin) into Hydroxyapatite Samples: Characterization and In Vitro Biological Evaluation.

Biomaterial-based bone grafts have an important role in the field of bone tissue engineering. One of the most promising classes of biomaterials is collagen, including the ones from marine biodiversity (in general, called spongin (SPG)). Also, hydroxyapatite (HA) has an important role in stimulating bone metabolism. Therefore, this work investigated the association of HA and SPG composites in order to evaluate their physico-chemical and morphological characteristics and their in vitro biological performance. For this, pre-set composite disks were evaluated by means of mass loss after incubation, pH, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and "in vitro" cell viability. pH measurements showed no statistical difference between groups. Moreover, a higher mass loss was observed for HA/SPG70/30 compared to the other groups for all experimental periods. Moreover, SEM representative micrographs showed the degradation of the samples with and without immersion. FTIR analysis demonstrated the absorption peaks for poly(methyl methacrylate) (PMMA), HA, and SPG. A higher L292 cell viability for control and PMMA was observed compared to HA and HA/SPG 90/10. Also, HA/SPG 70/30 showed higher cell viability compared to HA and HA/SPG 90/10 on days 3 and 7 days of culture. Furthermore, HA showed a significant lower MC3T3 cell viability compared to control and HA/SPG 70/30 on day 3 and no significant difference was observed between the composites in the last experimental period. Based on our investigations, it can be concluded that the mentioned composites were successfully obtained, presenting improved biological properties, especially the one mimicking the composition of bone (with 70% of HA and 30% of SPG). Consequently, these data highlight the potential of the introduction of SPG into HA to improve the performance of the graft for bone regeneration applications. Further long-term studies should be carried out to provide additional information concerning the late stages of material degradation and bone healing in the presence of HA/SPG.

Characterization and Cytotoxicity Evaluation of a Marine Sponge Biosilica.

Bone fractures characterize an important event in the medical healthcare, being related to traumas, aging, and diseases. In critical conditions, such as extensive bone loss and osteoporosis, the tissue restoration may be compromised and culminate in a non-union consolidation. In this context, the osteogenic properties of biomaterials with a natural origin have gained prominence. Particularly, marine sponges are promising organisms that can be exploited as biomaterials for bone grafts. Thus, the objectives of this study were to study the physicochemical and morphological properties of biosilica (BS) from sponges by using scanning electron microscopy, Fourier-transform infrared, X-ray diffraction (SEM, FTIR and XRD respectively), mineralization, and pH. In addition, tests on an osteoblast precursor cell line (MC3T3-E1) were performed to investigate its cytotoxicity and proliferation in presence of BS. Bioglass (BG) was used as gold standard material for comparison purposes. Sponge BS was obtained, and this fact was proven by SEM, FTIR, and XRD analysis. Calcium assay showed a progressive release of this ion from day 7 and a more balanced pH for BS was maintained compared to BG. Cytotoxicity assay indicated that BS had a positive influence on MC3T3-E1 cells viability and qRT-PCR showed that this material stimulated Runx2 and BMP4 gene expressions. Taken together, the results indicate a potential use of sponge biosilica for tissue engineering applications.

Characterization and biocompatibility of a fibrous glassy scaffold.

Bioactive glasses (BGs) are known for their ability to bond to living bone and cartilage. In general, they are readily available in powder and monolithic forms, which are not ideal for the optimal filling of bone defects with irregular shapes. In this context, the development of BG-based scaffolds containing flexible fibres is a relevant approach to improve the performance of BGs. This study is aimed at characterizing a new, highly porous, fibrous glassy scaffold and evaluating its in vitro and in vivo biocompatibility. The developed scaffolds were characterized in terms of porosity, mineralization and morphological features. Additionally, fibroblast and osteoblast cells were seeded in contact with extracts of the scaffolds to assess cell proliferation and genotoxicity after 24, 72 and 144 h. Finally, scaffolds were placed subcutaneously in rats for 15, 30 and 60 days. The scaffolds presented interconnected porous structures, and the precursor bioglass could mineralize a hydroxyapatite (HCA) layer in simulated body fluid (SBF) after only 12 h. The biomaterial elicited increased fibroblast and osteoblast cell proliferation, and no DNA damage was observed. The in vivo experiment showed degradation of the biomaterial over time, with soft tissue ingrowth into the degraded area and the presence of multinucleated giant cells around the implant. At day 60, the scaffolds were almost completely degraded and an organized granulation tissue filled the area. The results highlight the potential of this fibrous, glassy material for bone regeneration, due to its bioactive properties, non-cytotoxicity and biocompatibility. Future investigations should focus on translating these findings to orthotopic applications. Copyright © 2015 John Wiley & Sons, Ltd.

Characterization and biological evaluation of the introduction of PLGA into biosilicate®.

The aims of this study were to characterize different BS/PLGA composites for their physicochemical and morphological characteristics and evaluate the in vitro and in vivo biological performance. The physicochemical and morphological modifications were analyzed by pH, mass loss, XRD, setting time, and SEM. For in vitro analysis, the osteoblast and fibroblast viability was evaluated. For in vivo evaluations, histopathology and immunohistochemistry were performed in a tibial defect in rats. After incubation, all composites presented lower values in pH and mass loss over time. Moreover, XRD and SEM analysis confirmed that the composites degraded over time. Additionally, pore formation was observed by SEM analysis after incubation mainly in BS/PLGA groups. BS/PLGA showed significantly increased in osteoblast viability 24 h. Moreover, BS/PLGA composites demonstrated an increase in fibroblast viability in all periods analyzed when compared to BS. In the in vivo study, after 2 and 6 weeks of implantation of biomaterials, histopathological findings revealed that the BS/PLGA composites degrades over time, mainly at periphery. Moreover, can be observed the presence of granulation tissue, bone formation, Runx-2, and RANKL immunoexpression in all groups. In conclusion, BS/PLGA composites present appropriate physicochemical characteristics, stimulate the cellular viability, and enhance the bone repair in vivo. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1063-1074, 2017.

Effects of photobiomodulation on the fatigue level in elderly women: an isokinetic dynamometry evaluation.

Aging is responsible by a series of morphological and functional modifications that lead to a decline of muscle function, particularly in females. Muscle tissue in elderly people is more susceptible to fatigue and, consequently, to an increased inability to maintain strength and motor control. In this context, therapeutic approaches able of attenuating muscle fatigue have been investigated. Among these, the photobiomodulation demonstrate positive results to interacts with biological tissues, promoting the increase in cell energy production. Thus, the aim of this study was to investigate the effects of photobiomodulation (808 nm, 250 J/cm(2), 100 mW, 7 J each point) in the fatigue level and muscle performance in elderly women. Thirty subjects entered a crossover randomized double-blinded placebo-controlled trial. Photobiomodulation was delivered on the rectus femoris muscle of the dominant limb immediately before the fatigue protocol. In both sessions, peripheral muscle fatigue was analyzed by surface electromyography (EMG) and blood lactate analysis. Muscle performance was evaluated using an isokinetic dynamometer. The results showed that photobiomodulation was able of reducing muscle fatigue by a significant increase of electromyographic fatigue index (EFI) (p = 0.047) and decreasing significantly lactate concentration 6 min after the performance of the fatigue protocol (p = 0. 0006) compared the placebo laser session. However, the photobiomodulation was not able of increasing muscle performance measured by the isokinetic dynamometer. Thus, it can be conclude that the photobiomodulation was effective in reducing fatigue levels. However, no effects of photobiomodulation on muscle performance was observed.

Effect of three exercise programs on patients with chronic obstructive pulmonary disease.

We compared the effect of three different exercise programs on patients with chronic obstructive pulmonary disease including strength training at 50_80% of one-repetition maximum (1-RM) (ST; N = 11), low-intensity general training (LGT; N = 13), or combined training groups (CT; N = 11). Body composition, muscle strength, treadmill endurance test (TEnd), 6-min walk test (6MWT), Saint George's Respiratory Questionnaire (SGRQ), and baseline dyspnea (BDI) were assessed prior to and after the training programs (12 weeks). The training modalities showed similar improvements (P > 0.05) in SGRQ-total (ST = 13 +/- 14%; CT = 12 +/- 14%; LGT = 11 +/- 10%), BDI (ST = 1.8 +/- 4; CT = 1.8 +/- 3; LGT = 1 +/- 2), 6MWT (ST = 43 +/- 51 m; CT = 48 +/- 50 m; LGT = 31 +/- 75 m), and TEnd (ST = 11 +/- 20 min; CT = 11 +/- 11 min; LGT = 7 +/- 5 min). In the ST and CT groups, an additional improvement in 1-RM values was shown (P < 0.05) compared to the LGT group (ST = 10 +/- 6 to 57 +/- 36 kg; CT = 6 +/- 2 to 38 +/- 16 kg; LGT = 1 +/- 2 to 16 +/- 12 kg). The addition of strength training to our current training program increased muscle strength; however, it produced no additional improvement in walking endurance, dyspnea or quality of life. A simple combined training program provides benefits without increasing the duration of the training sessions.

Effect of three exercise programs on patients with chronic obstructive pulmonary disease

We compared the effect of three different exercise programs on patients with chronic obstructive pulmonary disease including strength training at 50_80 percent of one-repetition maximum (1-RM) (ST; N = 11), low-intensity general training (LGT; N = 13), or combined training groups (CT; N = 11). Body composition, muscle strength, treadmill endurance test (TEnd), 6-min walk test (6MWT), Saint George's Respiratory Questionnaire (SGRQ), and baseline dyspnea (BDI) were assessed prior to and after the training programs (12 weeks). The training modalities showed similar improvements (P > 0.05) in SGRQ-total (ST = 13 ± 14 percent; CT = 12 ± 14 percent; LGT = 11 ± 10 percent), BDI (ST = 1.8 ± 4; CT = 1.8 ± 3; LGT = 1 ± 2), 6MWT (ST = 43 ± 51 m; CT = 48 ± 50 m; LGT = 31 ± 75 m), and TEnd (ST = 11 ± 20 min; CT = 11 ± 11 min; LGT = 7 ± 5 min). In the ST and CT groups, an additional improvement in 1-RM values was shown (P < 0.05) compared to the LGT group (ST = 10 ± 6 to 57 ± 36 kg; CT = 6 ± 2 to 38 ± 16 kg; LGT = 1 ± 2 to 16 ± 12 kg). The addition of strength training to our current training program increased muscle strength; however, it produced no additional improvement in walking endurance, dyspnea or quality of life. A simple combined training program provides benefits without increasing the duration of the training sessions.

Pulmonary function testing in quadriplegic subjects.

STUDY DESIGN: Cross-sectional study. OBJECTIVE: Pulmonary functional capacity in 23 Brazilian quadriplegic subjects (ASIA A), aged 30 (9.5) years, weight 66 (10.75) kg, height 176 (7) cm, was investigated at 42 (64) months postinjury. SETTING: University Hospital--UNICAMP, Campinas, Brazil. METHOD: Subjects performed forced vital capacity (FVC) and maximal voluntary ventilation (MVV) tests while seated in their standard wheelchairs. Forced Expired Volume after 1 s (FEV1) and FVC/FEV1 ratio were calculated from these tests. Values obtained were compared to three prediction equations from the literature that are used specifically for spinal cord subjects and include different variables in their formulae, such as age, gender, height, postinjury time and injury level. Data are expressed as median (interquartile interval). Differences between values were demonstrated by median confidence interval with significance level set at alpha=0.05. RESULTS: Obtained data were statistically different from prediction equation results, with FVC 3.11 (0.81), 4.46 (0.28), 4.16 (0.33), 4.26 (0.42); FEV1 2.77 (1.03), 3.67 (0.21), 3.66 (0.30), 3.45 (0.39) and MVV 92 (27), 154.2 (11.9), 156.6 (14), 157.3 (16.8), where the first value is obtained experimentally and the second, third and fourth values correspond to predicted values. The results obtained from spirometry test in this study differed significantly from the results obtained when prediction equations were used. CONCLUSION: The use of prediction equations developed to estimate pulmonary function in wheelchair users significantly overestimates pulmonary function of quadriplegic individuals with complete lesions (ASIA group A), in comparison to measured values.

The effects of laser irradiation on osteoblast and osteosarcoma cell proliferation and differentiation in vitro.

OBJECTIVE: The aim of this study was to investigate the effects of 670-nm, 780-nm, and 830-nm laser irradiation on cell proliferation of normal primary osteoblast (MC3T3) and malignant osteosarcoma (MG63) cell lines in vitro. BACKGROUND: Some studies have shown that laser phototherapy is able to stimulate the osteogenesis of bone tissue, increasing osteoblast proliferation and accelerating fracture consolidation. It has been suggested that laser light may have a biostimulatory effect on tumor cells. However, the mechanism by which the laser acts on cells is not fully understood. MATERIALS AND METHODS: Neonatal, murine, calvarial, osteoblastic, and human osteosarcoma cell lines were studied. A single laser irradiation was performed at three different wavelengths, at the energies of 0.5, 1, 5, and 10 J/cm(2). Twenty-four hours after laser irradiation, cell proliferation and alkaline phosphatase assays were assessed. RESULTS: Osteoblast proliferation increased significantly after 830-nm laser irradiation (at 10 J/cm(2)) but decreased after 780-nm laser irradiation (at 1, 5, and 10 J/cm(2)). Osteosarcoma cell proliferation increased significantly after 670-nm (at 5 J/cm(2)) and 780-nm laser irradiation (at 1, 5, and 10 J/cm(2)), but not after 830-nm laser irradiation. Alkaline phosphatase (ALP) activity in the osteoblast line was increased after 830-nm laser irradiation at 10 J/cm(2), whereas ALP activity in the osteosarcoma line was not altered, regardless of laser wavelength or intensity. CONCLUSION: Based on the conditions of this study, we conclude that each cell line responds differently to specific wavelength and dose combinations. Further investigations are required to investigate the physiological mechanisms responsible for the contrasting outcomes obtained when using laser irradiation on cultured normal and malignant bone cells.

Efeitos de um programa de atividade física na postura hipercifótica torácica, na dorsalgia e na qualidade de vida de mulheres com osteoporose / Effects of a physical exercise program on the thoracic hyperkyphosis posture, on back pain and on quality of life in osteoporotic women

O objetivo deste estudo foi avaliar os efeitos de um programa de exercicios na postura hipercifotica toracica, na dorsalgia e na qualidade de vida dde mulhres com osteoporose. Sujeitos: 16 mulheres, com idade entre 65 e 75 anos. Os voluntarios foram submetidos a uma avaliacao fisica e a uma avaliacao do grau de cifose e responderam a questionarios para avaliar a dorsalgia, o Br-MPQ, e a qualidade de vida, o OPAQ. O programa foi composto de tres sessoes por semana (com 1 hora de duracao cada), durante 12 semanas. Os dados foram analisados estatisticamente pelo teste nao-parametrico de Wilcoxon. Foram observadas diferencas significativas(p=0,05) no grau de cifose (de 58,19o. +-14,83o. na avaliacao para 55o. +- 13,26o. na reavaliacao), diminuicao significativa na dorsalgia e melhora significativa da qualidade de vida. Os resultados encontrados permitem concluir que o programa de exercicios foi eficiente para melhorar a postura (com diminuicao do grau cifose), reduzir a dorsalgia e melhorar a qualidade de vida das mulheres osteoporoticas. Este estudo atingiu seu objetivo de desenvolver um programa de exercicios eficiente para melhorar alguns dos problemas fisicos presentes em individuos osteoporoticos