Bioactive glass-ceramic for bone tissue engineering: an in vitro and in vivo study focusing on osteoclasts

Abstract: Despite the crucial role of osteoclasts in the physiological process of bone repair, most bone tissue engineering strategies have focused on osteoblast-biomaterial interactions. Although Biosilicate® with two crystalline phases (BioS-2P) exhibits osteogenic properties and significant bone formation, its effects on osteoclasts are unknown. This study aimed to investigate the in vitro and in vivo effects of BioS-2P on osteoclast differentiation and activity. RAW 264.7 cells were cultured in osteoclastogenic medium (OCM) or OCM conditioned with BioS-2P (OCM-BioS-2P), and the cell morphology, viability, and osteoclast differentiation were evaluated. BioS-2P scaffolds were implanted into rat calvarial defects, and the bone tissue was evaluated using tartrate-resistant acid phosphatase (TRAP) staining and RT-polymerase chain reaction (PCR) after 2 and 4 weeks to determine the gene expressions of osteoclast markers and compare them with those of the bone grown in empty defects (Control). OCM-BioS-2P favored osteoclast viability and activity, as evidenced by an increase in the TRAP-positive cells and matrix resorption. The bone tissue grown on BioS-2P scaffolds exhibited higher expression of the osteoclast marker genes (Ctsk, Mmp 9, Rank) after 2 and 4 weeks and the RankL/Opg ratio after 2 weeks. Trap gene expression was lower at 2 weeks, and a higher number of TRAP-stained areas were observed in the newly formed bone on BioS-2P scaffolds at both 2 and 4 weeks compared to the Controls. These results enhanced our understanding of the role of bioactive glass-ceramics in bone repair, and highlighted their role in the modulation of osteoclastic activities and promotion of interactions between bone tissues and biomaterials.

In Vitro Effect of Low-Level Laser Therapy on Undifferentiated Mouse Pulp Cells / Efeito in Vitro da Laserterapia de Baixa Intensidade em Células Pulpares Indiferenciadas de Camundongo

Low-level laser therapy has been investigated as a possible stimulus for enhancement of proliferation and differentiation of various cell types, but few reports relate undifferentiated mouse pulp cells (OD-21) response to irradiation in in vitro models. The aim of this study was to analyze the influence of low-level laser therapy (λ=660 nm), with three different irradiation times, on the behavior of OD-21 cell line. The cells were cultivated and divided into three groups: non-irradiated/control (group I); irradiated with 88 s (group II); irradiated with 177 s (group III) and irradiated with 265 s (group IV). Cell growth and viability were assessed after 7 and 10 days. Data were analyzed by Kruskal-Wallis and MannWhitney tests (α=.05). At day 7, there was a higher cell growth in groups I and II, as compared to group IV (p<.01). At the 10th day, group I showed a higher cell growth as compared to group II (p<.05). Cell viability in group IV was significantly lower at the 7th day, as compared to groups I (p<.001), II (p<.01) and III (p<.001). Cell viability in all the groups was over 80%, except in group IV at day 7. Irradiation time of group I influenced positively the proliferation and viability of OD-21 cells in late cell culture period. (AU)

A terapia a laser de baixa intensidade tem sido investigada como possível estímulo para aumento da proliferação e diferenciação de vários tipos de células, mas poucos relatos relacionam a resposta de células indiferenciadas da polpa dentária de camundongos (OD-21) à irradiação em modelos in vitro. O objetivo deste estudo foi analisar a influência do laser de baixa intensidade (λ=660 nm), com três períodos de irradiação diferentes, no comportamento das células da linhagem OD-21. As células foram cultivadas e distribuídas em três grupos: não irradiado / controle (grupo I); irradiado com 88 s (grupo II); irradiado com 177 s (grupo III) e irradiado com 265 s (grupo IV). O crescimento e a viabilidade celular foram avaliados após 7 e 10 dias. Os dados foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney (α = 0,05). No dia 7, houve crescimento celular maior nos grupos I e II, em comparação ao grupo IV (p <0,01). No décimo dia, o grupo I apresentou crescimento celular superior ao grupo II (p <0,05). A viabilidade celular no grupo IV foi significativamente menor no sétimo dia, em comparação aos grupos I (p <0,001), II (p <0,01) e III (p <0,001). A viabilidade celular em todos os grupos foi superior a 80%, exceto no grupo IV no dia 7. O tempo de irradiação do grupo I influenciou positivamente a proliferação e a viabilidade das células OD-21 no período mais tardio da cultura celular.A terapia a laser de baixa intensidade tem sido investigada como possível estímulo para aumento da proliferação e diferenciação de vários tipos de células, mas poucos relatos relacionam a resposta de células indiferenciadas da polpa dentária de camundongos (OD-21) à irradiação em modelos in vitro. O objetivo deste estudo foi analisar a influência do laser de baixa intensidade (λ=660 nm), com três períodos de irradiação diferentes, no comportamento das células da linhagem OD-21. As células foram cultivadas e distribuídas em três grupos: não irradiado / controle (grupo I); irradiado com 88 s (grupo II); irradiado com 177 s (grupo III) e irradiado com 265 s (grupo IV). O crescimento e a viabilidade celular foram avaliados após 7 e 10 dias. Os dados foram analisados pelos testes de Kruskal-Wallis e Mann-Whitney (α = 0,05). No dia 7, houve crescimento celular maior nos grupos I e II, em comparação ao grupo IV (p <0,01). No décimo dia, o grupo I apresentou crescimento celular superior ao grupo II (p <0,05). A viabilidade celular no grupo IV foi significativamente menor no sétimo dia, em comparação aos grupos I (p <0,001), II (p <0,01) e III (p <0,001). A viabilidade celular em todos os grupos foi superior a 80%, exceto no grupo IV no dia 7. O tempo de irradiação do grupo I influenciou positivamente a proliferação e a viabilidade das células OD-21 no período mais tardio da cultura celular. (AU)

Effect of focal adhesion kinase inhibition on osteoblastic cells grown on titanium with different topographies

Abstract Objective The present study aimed to investigate the participation of focal adhesion kinases (FAK) in interactions between osteoblastic cells and titanium (Ti) surfaces with three different topographies, namely, untreated (US), microstructured (MS), and nanostructured (NS). Methodology Osteoblasts harvested from the calvarial bones of 3-day-old rats were cultured on US, MS and NS discs in the presence of PF-573228 (FAK inhibitor) to evaluate osteoblastic differentiation. After 24 h, we evaluated osteoblast morphology and vinculin expression, and on day 10, the following parameters: gene expression of osteoblastic markers and integrin signaling components, FAK protein expression and alkaline phosphatase (ALP) activity. A smooth surface, porosities at the microscale level, and nanocavities were observed in US, MS, and NS, respectively. Results FAK inhibition decreased the number of filopodia in cells grown on US and MS compared with that in NS. FAK inhibition decreased the gene expression of Alp, bone sialoprotein, osteocalcin, and ALP activity in cells grown on all evaluated surfaces. FAK inhibition did not affect the gene expression of Fak, integrin alpha 1 ( Itga1 ) and integrin beta 1 ( Itgb1 ) in cells grown on MS, increased the gene expression of Fak in cells grown on NS, and increased the gene expression of Itga1 and Itgb1 in cells grown on US and NS. Moreover, FAK protein expression decreased in cells cultured on US but increased in cells cultured on MS and NS after FAK inhibition; no difference in the expression of vinculin was observed among cells grown on all surfaces. Conclusions Our data demonstrate the relevance of FAK in the interactions between osteoblastic cells and Ti surfaces regardless of surface topography. Nanotopography positively regulated FAK expression and integrin signaling pathway components during osteoblast differentiation. In this context, the development of Ti surfaces with the ability to upregulate FAK activity could positively impact the process of implant osseointegration.

Effect of stem cells combined with a polymer/ceramic membrane on osteoporotic bone repair

Abstract Cell therapy associated with guided bone regeneration (GBR) can be used to treat bone defects under challenging conditions such as osteoporosis. This study aimed to evaluate the effect of mesenchymal stem cells (MSCs) in combination with a poly(vinylidene-trifluoroethylene)/barium titanate (PVDF-TrFE/BT) membrane on bone repair in osteoporotic rats. Osteoporosis was induced in female rats by bilateral removal of the ovaries (OVX) or sham surgery (SHAM), and the osteoporotic condition was characterized after 5 months by microtomographic and morphometric analyses. Calvarial defects were created in osteoporotic rats that immediately received the PVDF-TrFE/BT membrane. After 2 weeks, bone marrow-derived MSCs from healthy rats, characterized by the expression of surface markers using flow cytometry, or phosphate-buffered saline (PBS) (Control) were injected into the defects and bone formation was evaluated 4 weeks post-injection by microtomographic, morphometric, and histological analyses. A reduction in the amount of bone tissue in the femurs of OVX compared with SHAM rats confirmed the osteoporotic condition of the experimental model. More bone formation was observed when the defects were injected with MSCs compared to that with PBS. The modification that we are proposing in this study for the classical GBR approach where cells are locally injected after a membrane implantation may be a promising therapeutic strategy to increase bone formation under osteoporotic condition.

Aging impairs osteoblast differentiation of mesenchymal stem cells grown on titanium by favoring adipogenesis

ABSTRACT Aging negatively affects bone/titanium implant interactions. Our hypothesis is that the unbalance between osteogenesis and adipogenesis induced by aging may be involved in this phenomenon. Objective We investigated the osteoblast and adipocyte differentiation of mesenchymal stem cells (MSCs) from young and aged rats cultured on Ti. Material and Methods Bone marrow MSCs derived from 1-month and 21-month rats were cultured on Ti discs under osteogenic conditions for periods of up to 21 days and osteoblast and adipocyte markers were evaluated. Results Cell proliferation, alkaline phosphatase (ALP) activity, extracellular matrix mineralization and gene expression of RUNX2, osterix, ALP, bone sialoprotein, osteopontin, and osteocalcin were reduced in cultures of 21-month rats compared with 1-month rats grown on Ti. Gene expression of PPAR-γ , adipocyte protein 2, and resistin and lipid accumulation were increased in cultures of 21-month rats compared with 1-month rats grown on the same conditions. Conclusions These results indicate that the lower osteogenic potential of MSCs derived from aged rats compared with young rats goes along with the higher adipogenic potential in cultures grown on Ti surface. This unbalance between osteoblast and adipocyte differentiation should be considered in dental implant therapy to the elderly population.

Effect of collagen sponge and fibrin glue on bone repair

ABSTRACT The ability of hemostatic agents to promote bone repair has been investigated using in vitro and in vivo models but, up to now, the results are inconclusive. Objective In this context, the aim of this study was to compare the potential of bone repair of collagen sponge with fibrin glue in a rat calvarial defect model. Material and Methods Defects of 5 mm in diameter were created in rat calvariae and treated with either collagen sponge or fibrin glue; untreated defects were used as control. At 4 and 8 weeks, histological analysis and micro-CT-based histomorphometry were carried out and data were compared by two-way ANOVA followed by Student-Newman-Keuls test when appropriated (p≤0.05). Results Three-dimensional reconstructions showed increased bone formation in defects treated with either collagen sponge or fibrin glue compared with untreated defects, which was confirmed by the histological analysis. Morphometric parameters indicated the progression of bone formation from 4 to 8 weeks. Additionally, fibrin glue displayed slightly higher bone formation rate when compared with collagen sponge. Conclusion Our results have shown the benefits of using collagen sponge and fibrin glue to promote new bone formation in rat calvarial bone defects, the latter being discreetly more advantageous.

Bone tissue response to plasma-nitrided titanium implant surfaces

A current goal of dental implant research is the development of titanium (Ti) surfaces to improve osseointegration. Plasma nitriding treatments generate surfaces that favor osteoblast differentiation, a key event to the process of osteogenesis. Based on this, it is possible to hypothesize that plasma-nitrided Ti implants may positively impact osseointegration. Objective The aim of this study was to evaluate the in vivo bone response to Ti surfaces modified by plasma-nitriding treatments. Material and Methods Surface treatments consisted of 20% N2 and 80% H2, 450°C and 1.5 mbar during 1 h for planar and 3 h for hollow cathode. Untreated surface was used as control. Ten implants of each surface were placed into rabbit tibiae and 6 weeks post-implantation they were harvested for histological and histomorphometric analyses. Results Bone formation was observed in contact with all implants without statistically significant differences among the evaluated surfaces in terms of bone-to-implant contact, bone area between threads, and bone area within the mirror area. Conclusion Our results indicate that plasma nitriding treatments generate Ti implants that induce similar bone response to the untreated ones. Thus, as these treatments improve the physico-chemical properties of Ti without affecting its biocompatibility, they could be combined with modifications that favor bone formation in order to develop new implant surfaces. .

Oxidative nanopatterning of titanium surfaces promotes production and extracellular accumulation of osteopontin

The bone-biomaterial interface has been characterized by layers of afibrillar extracellular matrix (ECM) enriched in non collagenous proteins, including osteopontin (OPN), a multifunctional protein that in bone controls cell adhesion and ECM mineralization. Physical and chemical aspects of biomaterial surfaces have been demonstrated to affect cell-ECM-substrate interactions. The present paper described the ability of oxidative nanopatterning of titanium (Ti) surfaces to control extracellular OPN deposition in vitro. Ti discs were chemically treated by a mixture of H2SO4/H2O2 for either 30 min [Nano(30') Ti] or 4 h [Nano(4h) Ti]. Non-etched Ti discs were used as control. Primary osteogenic cells derived from newborn rat calvarial bone were plated on control and etched Ti and grown under osteogenic conditions up to 7 days. High resolution scanning electron microscopy revealed that treated Ti discs exhibited a nanoporous surface and that areas of larger nanopits were noticed only for Nano(4h) Ti. Large extracellular OPN accumulation were detectable only for Nano(4h) Ti, which was associated with OPN-positive cells with typical aspects of migrating cells. At day 3, quantitative results in terms of areas of OPN labeling were as follows: Nano(4h) Ti > Nano(30') Ti > Control Ti. In conclusion, chemically nanostructured Ti surfaces may support the enhancement of endogenous extracellular OPN deposition by osteogenic cells in vitro depending on the etching time, a finding that should be taken into consideration in strategies to biofunctionalize implant surfaces with molecules with cell adhesion capacity.

A interface osso-implante é caracterizada pela presença de uma camada de matriz extracellular (MEC) afibrilar rica em proteínas não-colágenas, incluindo osteopontina (OPN), cujas funções no tecido ósseo estão relacionadas à adesão celular e ao controle do processo de mineralização da MEC (crescimento de cristais). Aspectos físicos e químicos das superfícies de biomateriais podem afetar as interações célula-MEC-substrato. O objetivo do presente estudo foi demonstrar a capacidade de aspectos nanotopográficos de superfície de titânio (Ti) de controlar a deposição extracelular de OPN in vitro. Discos de Ti foram tratados quimicamente por solução de H2SO4/H2O2 durante 30 min [Nano(30') Ti] ou 4 h [Nano(4h) Ti]. Superfícies de Ti não tratadas foram usadas como controle. Células osteogênicas primárias derivadas de calvárias de ratos recém-nascidos foram plaqueadas sobre os discos de Ti e cultivadas em condições osteogênicas por até 7 dias. Microscopia eletrônica de varredura de alta resolução revelou que os discos de Ti tratados quimicamente exibiam superfície nanoporosa, com áreas de nanoporos maiores para Nano(4h) Ti. Apenas para esse grupo detectavam-se acúmulos extensos de OPN extracelular, os quais se distribuíam em áreas adjacentes a células OPN-positivas, com aspectos morfológicos típicos de células em migração. Em conclusão, a nanoestruturação química de superfície de Ti pode favorecer o aumento da deposição extracelular de OPN endógena por células osteogênicas in vitro, dependendo do tempo de condicionamento utilizado, o que deve ser considerado no desenvolvimento de estratégias para funcionalizar superfícies de implantes com moléculas com reconhecido efeito no processo de adesão celular.

Effects of a novel calcium aluminate cement on the early events of the progressionof osteogenic cell cultures

The present study evaluated the progression of osteogenic cell cultures exposed to a novel calcium aluminate cement (CAC+) in comparison with the gold standard mineral trioxide aggregate (MTA). Cells were enzimatically isolated from newborn rat calvarial bone, plated on glass coverslips containing either CAC+ or a control MTA samples in the center, and grown under standard osteogenic conditions. Over the 10-day culture period, roundening of sample edges was clearly noticed only for MTA group. Although both cements supported osteogenic cell adhesion, spreading, and proliferation, CAC+-exposed cultures showed significantly higher values in terms of total cell number at days 3 and 7, and total protein content and alkaline phosphatase activity at day 10. The present in vitro results indicate that the exposure to CAC+ supports a higher differentiation of osteogenic cells compared with the ones exposed to MTA. Further experimental studies should consider CAC+ as a potential alternative to MTA when the repair of mineralized tissues is one of the desired outcomes in endodontic therapy.

O objetivo do presente estudo foi avaliar a progressão de cultura de células osteogênicas expostas a um novo cimento de aluminato de cálcio (CAC+) em comparação ao agregado de trióxido mineral (MTA). As células foram obtidas por digestão enzimática de calvária de ratos recém-nascidos, plaqueadas sobre lamínulas de vidro contendo em sua área central discos de CAC+ ou MTA e crescidas em condições osteogênicas por até 10 dias. Durante a cultura primária, observou-se o arredondamento das bordas das amostras de cimento apenas para MTA. Embora ambos os cimentos tenham permitido a adesão, o espraiamento e a proliferação celulares, as culturas crescidas em contato com CAC+ exibiram valores maiores de número total de células em 3 e 7 dias, e de conteúdo de proteína total e atividade de fosfatase alcalina em 10 dias. Os resultados indicam que a exposição ao CAC+ permite o desenvolvimento de uma proporção maior de células em estágios mais avançados da diferenciação osteoblástica, quando comparado ao MTA. Deve-se considerar em futuros estudos experimentais a utilização do CAC+ como um material alternativo ao MTA especialmente quando um dos objetivos do tratamento endodôntico é o de reparação dos tecidos mineralizados da região periapical.

Paralisia facial após técnica anestésica mandibular / Facial nerve palsy after mandibular anesthesia technique

A paralisia do nervo facial tem múltiplas etiologias, como viroses, traumatismo facial, codutas iatrogênicas, tumores, origem idiopática, infarto cerebral e paralisia pseudobulbar, sendo, portanto, um achado bastante raro durante o tratamento dentário. Nessas situações, a paralisia pode estar associada à realização errônea da técnica anestésica, a procedimentos cirúrgicos prolongados de extração dentária, ou ainda, a infecções de origem dental. Neste trabalho, é demonstrado um caso de paralisia hemi-facial completa, ocorrida durante anestesia dos nervos lingual, alveolar inferior e bucal em uma paciente de 63 anos. A recuperação completa dos movimentos faciais foi observada após algumas horas, com a paciente sendo liberada, e o procedimento cirúrgico realizado duas semanas mais tarde.

Facial nerve palsy has many etiologies, such as viruses, facial trauma, iatrogenesis, tumors, idiopathic conditions, cerebral infarction and pseudobulbar palsy, as a result of which it is rarely observed during dental treatment. In this situation, it may be associated with the injection of a local anesthetic, prolonged surgical procedure for the removal of mandibular molars and infections of dental origin. We report a case of complete facial nerve palsy during a mandibular nerve block anesthesia in a 63-year-old woman. The full recovery of facial movements was attained after a few hours and she was discharged. The surgical procedure was performed two weeks later.

Characterization and in vitro cytocompatibility of an acid-etched titanium surface

The aims of this study were to characterize the microstructure of a commercially pure titanium (cpTi) surface etched with HCl/H2SO4 (AE-cpTi) and to investigate its in vitro cytocompatibility compared to turned cpTi (T-cpTi). T-cpTi showed a grooved surface and AE-cpTi revealed a surface characterized by the presence of micropits. Surface parameters indicated that the AE-cpTi surface is more isotropic and present a greater area compared to T-cpTi. The oxide film thickness was similar between both surfaces; however, AE-cpTi presented more Ti and O and less C. Osteoblastic cell proliferation, alkaline phosphatase activity, and bone-like nodule formation were greater on T-cpTi than on AE-cpTi. These results show that acid etching treatment produced a surface with different topographical and chemical features compared to the turned one, and such surface modification affected negatively the in vitro cytocompatibility of cpTi as demonstrated by decreasing culture growth and expression of osteoblastic phenotype.

O objetivo deste estudo foi caracterizar a microestrutura de uma superfície de titânio comercialmente puro (cpTi) condicionada com HCl/H2SO4 (acid etched) (AE-cpTi) e investigar sua citocompatibilidade in vitro, comparada à do cpTi usinado (turned) (T-cpTi). O T-cpTi apresentou uma superfície com sulcos e o AE-cpTi exibiu uma superfície caracterizada pela presença de micro-vales. Os parâmetros de superfície indicaram que a superfície AE-cpTi é mais isotrópica e apresenta uma área maior quando comparada à superfície T-cpTi. A espessura da camada de óxido foi similar para as duas superfícies; no entanto, a AE-cpTi apresentou maiores quantidades de Ti e O e menor, de C. A proliferação de células osteoblásticas, a atividade de fosfatase alcalina e a formação de matriz mineralizada foram maiores na superfície T-cpTi que na AE-cpTi. Esses resultados mostram que o condicionamento ácido produziu uma superfície com características topográficas e químicas diferentes quando comparadas às da superfície usinada. Além disso, observou-se que essas modificações de superfície afetaram de forma negativa a citocompatibilidade in vitro do cpTi como demonstrado pela inibição da proliferação celular e da expressão do fenótipo osteoblástico.

Effects of low-level laser therapy on human osteoblastic cells grown on titanium

The aim of this study was to investigate the effects of low-level laser therapy (LLLT) by using gallium aluminum arsenide (GaAlAs) diode laser on human osteoblastic cells grown on titanium (Ti). Osteoblastic cells were obtained by enzymatic digestion of human alveolar bone and cultured on Ti discs for up to 17 days. Cells were exposed to LLLT at 3 J/cm2 (wavelength of 780 nm) at days 3 and 7 and non-irradiated cultures were used as control. LLLT treatment did not influence culture growth, ALP activity, and mineralized matrix formation. Analysis of cultures by epifluorescence microscopy revealed an area without cells in LLLT treated cultures, which was repopulated latter with proliferative and less differentiated cells. Gene expression of ALP, OC, BSP, and BMP-7 was higher in LLLT treated cultures, while Runx2, OPN, and OPG were lower. These results indicate that LLLT modulates cell responses in a complex way stimulating osteoblastic differentiation, which suggests possible benefits on implant osseointegration despite a transient deleterious effect immediately after laser irradiation.

Este estudo teve como objetivo investigar o efeito do laser diodo de gálio-alumínio-arsênio (GaAlAs) em células osteoblásticas humanas cultivadas sobre discos de Ti. Para tanto, células osteoblásticas foram obtidas por digestão enzimática de osso alveolar humano e cultivadas sobre discos de Ti por 17 dias. As células foram submetidas à irradiação no 3º e 7º dias na dose de 3 J/cm2 e comprimento de onda de 780 nm e células não irradiadas foram usadas como controle. A irradiação não alterou a proliferação celular, atividade de ALP e formação de matriz mineralizada. Microscopia por epifluorescência indicou que após 24 h da aplicação do laser, as culturas irradiadas apresentaram áreas sem células, que mais tarde foram repovoadas por células em fase de proliferação e menos diferenciadas. O laser aumentou a expressão gênica relativa da ALP, OC, BSP e BMP-7 e reduziu a de RUNX2, OPN e OPG. Os resultados indicam que a terapia com laser modula de forma complexa as respostas celulares, estimulando a diferenciação osteoblástica. Assim, é possível sugerir possíveis benefícios do laser na osseointegração de implantes de Ti apesar do efeito deletério às células imediatamente após a irradiação.

Effect of temperature variation on the cytotoxicity of cast dental alloys and commercially pure titanium

Cell culture system has been used to evaluate alloy cytotoxicity under different environments, testing the extracts, but the effect of temperature variation on the cytotoxicity of dental alloys has not been analyzed. OBJECTIVE: The aim of the present study was to investigate if temperature variation could affect dental alloy cytotoxicity, testing alloy extracts in an epithelial cell culture system. MATERIAL AND METHODS: Discs of Ni-Cr, Co-Cr-Mo, Ni-Cr-Ti, Ti-6Al-4V and commercially pure titanium (cp Ti) were cast by arc melting, under argon atmosphere, injected by vacuum-pressure. Discs were immersed in artificial saliva and subjected to different temperatures: 37ºC and thermocycling (37ºC/5ºC/37ºC/55ºC/37ºC). After thermocycling, extracts were put in a subconfluent culture during 6 h, and the number of cells and their viability were used to evaluate cytotoxicity in these temperatures. For each alloy, data from temperature conditions were compared by Student's t-test (α=0.05). RESULTS: The cytotoxicity tests with alloy/metal extracts showed that Ni-Cr, Co-Cr-Mo, Ti-6Al-4V and cp Ti extracts (p>0.05) did not affect cell number or cell viability, while Ni-Cr-Ti (p<0.05) extract decreased cell number and viability when the alloy was subjected to thermocycling. CONCLUSION: Within the limitations of the present study, the Ni-Cr-Ti alloy had cell number and viability decreased when subjected to temperature variation, while the other alloys/metal extracts did not show these results.

Bone response to a Ca- and P-enriched titanium surface obtained by anodization

This study evaluated bone response to a Ca- and P- enriched titanium (Ti) surface treated by a multiphase anodic spark deposition coating (BSP-AK). Two mongrel dogs received bilateral implantation of 3 Ti cylinders (4.1 x 12 mm) in the humerus, being either BSP-AK treated or untreated (machined - control). At 8 weeks postimplantation, bone fragments containing the implants were harvested and processed for histologic and histomorphometric analyses. Bone formation was observed in cortical area and towards the medullary canal associated to approximately 1/3 of implant extension. In most cases, in the medullary area, collagen fiber bundles were detected adjacent and oriented parallel to Ti surfaces. Such connective tissue formation exhibited focal areas of mineralized matrix lined by active osteoblasts. The mean percentages of bone-to-implant contact were 2.3 (0.0-7.2 range) for BSP-AK and 0.4 (0.0-1.3 range) for control. Although the Mann-Whitney test did not detect statistically significant differences between groups, these results indicate a trend of BSP-AK treated surfaces to support contact osteogenesis in an experimental model that produces low bone-to-implant contact values.

O objetivo desse estudo foi avaliar a resposta do tecido ósseo à superfície de titânio (Ti) enriquecida com Ca e P obtida por anodização (BSP-AK). Três cilindros de Ti (4,1 x 12 mm) BSP-AK ou usinado (controle) foram implantados bilateralmente nos úmeros de dois cães de raça indefinida. Oito semanas após a implantação, os fragmentos ósseos contendo os implantes foram removidos e processados para análises histológica e histomorfométrica. A formação óssea foi observada na região cortical e no canal medular até aproximadamente um terço da extensão do implante. Na maioria dos casos, feixes de fibras colágenas dispostos paralelamente à superfície do implante foram observados na região medular. Nessa região observaram-se também áreas focais de formação de matriz mineralizada e osteoblastos ativos. Os implantes do grupo BSP-AK apresentaram média de contato osso-implante 2,3 por cento, com medidas variando de 0,0 a 7,2 por cento e os do grupo controle tiveram média 0,4 por cento, com medidas variando de 0,0 a 1,3 por cento. Apesar do teste de Mann-Whitney não mostrar diferença estatisticamente significante entre os grupos, nossos resultados indicaram uma tendência para a ocorrência de osteogênese de contato na superfície BSP-AK em um modelo experimental que resulta em baixos valores de contato osso-implante.

Electro-acupuncture efficacy on pain control after mandibular third molar surgery

The aim of this study was to evaluate the efficacy of electro-acupuncture (EAC) on postoperative pain control after mandibular third molar surgery. Twenty four young patients (12 male and 12 female) with symmetrically impacted mandibular third molars were selected. Each patient was submitted to two separate surgical procedures under local anesthesia. At one side, extraction was carried out employing both prior (24h) and immediately postoperative application of EAC, while on the contralateral side surgery was carried out without any treatment. EAC was applied on 6 bilateral systemic and 2 auricular points with a WQ10Dl appliance using 40-60Hz frequency for 20 min and individually adjusted intensity. Postoperative pain intensity was rated on a 100 mm visual analog scale (VAS) between 2 and 72 h and recording the amount of analgesics intake after surgery. Statistical analysis was performed using theWilcoxon test. Postoperative pain VAS scores were significantly lower for the EAC group (p<0.05) and analgesic intake decreased (p<0.05) for all evaluated periods (p<0.05). Under the tested conditions EAC therapy is efficient was proved controlling postoperative pain following mandibular third molar surgical removal.

O objetivo desta pesquisa foi verificar a eficácia da aplicação de eletro-acupuntura (EAC) na redução da dor após a exodontia de terceiros molares inferiores inclusos. Foram selecionados 24 pacientes jovens (12 homens e 12 mulheres) com inclusão bilateral de terceiros molares inferiores em posição similar. Cada paciente foi submetido aleatoriamente a dois procedimentos cirúrgicos em dias diferentes: em um deles o dente foi extraído com uma sessão pré-operatória (24 h) e uma pós-operatória imediata de EAC enquanto no outro a extração foi feita sem EAC. A EAC foi aplicada em 6 pontos sistêmicos bilaterais e 2 auriculares com um aparelho WQ10D1 utilizando freqüência de 40-60 Hz por 20 min com intensidade ajustada individualmente. A dor foi avaliada desde 2h até 72h pós-operatórias utilizando a escala visual análoga (EVA) de 100 mm e pelo consumo de analgésicos. Os dados foram comparados pelo teste deWilcoxon. Os escores de dor da EVA foram significantemente menores para o tratamento com EAC (p<0,05) enquanto o consumo de analgésico diminuiu (p<0,05) em todos os períodos (p<0,05). O tratamento com EAC mostrou-se eficiente no controle da dor pós-operatória após cirurgia de extração de terceiros molares inferiores inclusos.

Intraosseous schwannoma of mandibular symphysis: case report

Schwannoma (neurilemmoma) is a benign neoplasm originated from the neural sheath and occurring most frequently in the head and neck. Intraosseous schwannomas are rare. The mandible is the most common site of occurrence for these lesions. This article reports the case of an intraosseous schwannoma located in the mandibular symphysis of an 11-year-old boy. The lesion was surgically removed and no radiographic evidence of recurrence was observed after 5 years.

O schwannoma (neurilemoma) é um neoplasma benigno que se origina da bainha neural e ocorre mais freqüentemente na cabeça e pescoço. Schwannomas intra-ósseos são raros. O local mais comumente afetado é a mandíbula. Este artigo documenta um caso de schwannoma intra-ósseo localizado na sínfise mandibular de um menino com 11 anos de idade. A lesão foi removida cirurgicamente e não houve evidências radiográficas de recorrência após 5 anos de acompanhamento.

Development of the osteoblast phenotype of serial cell subcultures from human bone marrow

Culturas de células de medula óssea têm sido utilizadas para avaliar a biocompatibilidade de substitutos ósseos que poderiam ser empregados em cirurgias maxilofaciais e ortopédicas. No entanto, ainda não está claro se células em subculturas mantêm a habilidade para se diferenciarem em osteoblastos. O objetivo deste estudo foi comparar o desenvolvimento do fenótipo osteoblástico em subculturas seriadas de células de medula óssea humana. Células da primeira à terceira passagem foram cultivadas (2x104 células/poço) em a-MEM suplementado. As células foram incubadas a 37ºC e 5% CO2 / 95% ar atmosférico. A adesão celular foi avaliada em 4 h e 24 h. Aos 7, 14 e 21 dias, a proliferação e viabilidade celulares, conteúdo de proteína total e atividade de fosfatase alcalina (ALP) foram avaliadas. A formação de matriz mineralizada foi avaliada aos 14 e 21 dias. Os dados foram comparados por análise de variância a dois critérios e teste de Duncan. Adesão e viabilidade celular e conteúdo de proteína total não foram afetados pela subcultura seriada. Entretanto, a subcultura seriada interferiu negativamente na diferenciação osteoblástica, como demonstrado pelos parâmetros osteoblásticos da segunda e terceira passagens, tais como a proliferação celular contínua, a baixa atividade de ALP e a baixa quantidade de matriz mineralizada formada, quando comparados à primeira passagem. Portanto, é importante avaliar a capacidade das células para se diferenciarem em osteoblastos antes de selecionar a população de células adequada para testar a biocompatibilidade de materiais para substituir tecido ósseo.

Osteoblast differentiation of human bone marrow cells under continuous and discontinuous treatment with dexamethasone

A dexametasona (Dex) induz diferenciação osteoblástica em diversos modelos de cultura de células. Este estudo investigou o efeito do tratamento contínuo e descontínuo com Dex sobre a diferenciação de células de medula óssea humana (BMSC). Células da cultura primária e da primeira passagem foram cultivadas em meio de cultura com e sem Dex 10-7 M (37ºC e 5% CO2 / 95% ar atmosférico). Aos 7, 14 e 21 dias, os seguintes parâmetros foram avaliados: proliferação e viabilidade celulares, conteúdo de proteína total, atividade de fosfatase alcalina (ALP) e formação de matriz mineralizada. Os dados foram comparados por análise de variância a dois critérios. A Dex não afetou a viabilidade celular e o conteúdo de proteína total, mas reduziu o número de células. A atividade de ALP e a formação de matriz mineralizada foram aumentadas quando apenas a primeira passagem ou cultura primária e primeira passagem foram tratadas com Dex, em comparação aos grupos que não tiveram contato com Dex após a primeira passagem. Estes resultados indicam que, para BMSC humanas, a presença contínua de Dex não parece ser necessária para o desenvolvimento do fenótipo osteoblástico. Contudo, a Dex deve estar presente após a primeira passagem para permitir a diferenciação osteoblástica expressa por proliferação celular reduzida e aumento da atividade de ALP e da formação de matriz mineralizada.

Effect of cpTi surface roughness on human bone marrow cell attachment, proliferation, and differentiation

There is general agreement that rough surfaces improve both biologic and biomechanical responses to titanium (Ti) implants. The aim of this investigation was to study the effect of Ti surface roughness on the response of human bone marrow cell culture evaluating: cell attachment, cell proliferation, total protein content, alkaline phosphatase (ALP) activity, and bone-like nodule formation. Cells were cultured on commercially pure titanium (cpTi) discs with four different average roughnesses (Ra). For attachment evaluation, cells were cultured for 4 h. After 21 days, cell proliferation, total protein content, and ALP activity were evaluated. For bone-like nodule formation, cells were cultured for 28 days. Data were compared by ANOVA and Duncan's multiple range test. Cell attachment was not affected by surface roughness. For cells cultured on Ti with Ra ranging from 0.80 µm to 1.90 µm, proliferation was reduced while total protein content, and ALP activity were increased. There was a non-statistically significant increase of bone-like nodule formation on a surface with Ra near 0.80 µm. These results suggest that for Ti an Ra ranging from 0.80 µm to 1.90 µm would optimize both intermediary and final cellular responses but not affect the initial response, and a smoother surface would not favor any evaluated response

Surface topography of hydroxyapatite affects ROS17/2.8 cells response / A topografia de superfície da hidroxiapatita afeta a resposta de células ROS17/2.8

A hidroxiapatita (HA) tem sido utilizada como revestimento de implantes e para substituiçäo de tecido ósseo. O objetivo deste estudo foi avaliar o efeito da topografia de superfície da HA, resultante da presença de microporosidade, sobre a adesäo, morfologia e proliferaçäo celulares, a medida de proteína total e a atividade de fosfatase alcalina. Discos de HA com diferentes porcentagens de microporosidade (<5 por cento, 15 por cento e 30 por cento) foram fabricados por uma combina;äo das técnicas de pressäo uniaxial e sinterizaçäo. Células ROS172.8 foram cultivadas sobre os discos de HA. Para a adesäo, as células foram cultivadas por duas horas. A morfologia foi avaliada após sete dias. A proliferaçäo, medida de proteína total e atividade de ALP foram avaliadas após sete e quatorze dias. Os dados foram comparados por ANOVA e teste de Duncan quando apropriado. A adesäo (p=0,11) e a medida de proteína total (p=0,31) näo foram afetadas pela topografia de superfície. A proliferaçäo após sete e quatorze dias (p=0,0007 e p=0,003, respectivamente), e a atividade de ALP (p=0,0007) foram significantemente menores na superfície irregular (HA30). Esses resultados sugerem que eventos iniciais näo säo afetados pela topografia, enquanto superfícies com topografias mais regulares (microporosidade de 15 por cento ou menos) favoreceram eventos intermediários e finais, como proliferaçäo e atividade de ALP