Bone morphogenetic proteins (BMPs) are multi-functional growth factors belonging to the transforming growth factor ss superfamily. Family members are expressed during limb development, endochondral ossification, early fracture, and cartilage repair. The activity of BMPs was first identified in the 1960s but the proteins responsible for bone induction were unknown until the purification and cloning of human BMPs in the 1980s. To date, about 15 BMP family members have been identified and characterized. The signal triggered by BMPs is transduced through serine/threonine kinase receptors, type I and II subtypes. Three type I receptors have been shown to bind BMP ligands, namely: type IA and IB BMP receptors and type IA activin receptors. BMPs seem to be involved in the regulation of cell proliferation, survival, differentiation and apoptosis, but their hallmark is their ability to induce bone, cartilage, ligament, and tendon formation at both heterotopic and orthotopic sites. This suggests that, in the future, they may play a major role in the treatment of bone diseases. Several animal studies have illustrated the potential of BMPs to enhance spinal fusion, repair critical-size defects, accelerate union, and heal articular cartilage lesions. Difficulties in producing and purifying BMPs from bone tissue have prompted the attempts made by several laboratories, including ours, to express these proteins in the recombinant form in heterologous systems. This review focuses on BMP structure, molecular mechanisms of action and significance and potential applications in medical, dental and veterinary practice for the treatment of cartilage and bone-related diseases.
Titanium (Ti) and its alloys are widely used in dental implants and hip-prostheses due to their excellent biocompatibility. Growing evidence support that surface degradation due to corrosion and wear processes, contribute to implant failure, since the release of metallic ions and wear particles generate local tissue reactions (peri-implant inflammatory reactions). The generated ions and wear debris (particles at the micron and nanoscale) stay, in a first moment, at the interface implant-bone. However, depending on their size, they can enter blood circulation possibly contributing to systemic reactions and toxicities. Most of the nanotoxicological studies with titanium dioxide nanoparticles (TiO 2 NPs) use conventional two-dimensional cell culture monolayers to explore macrophage and monocyte activation, where limited information regarding bone cells is available. Recently three-dimensional models have been gaining prominence since they present a greater anatomical and physiological relevance. Taking this into consideration, in this work we developed a human osteoblast-like spheroid model, which closely mimics bone cell-cell interactions, providing a more realistic scenario for nanotoxicological studies. The treatment of spheroids with different concentrations of TiO 2 NPs during 72 h did not change their viability significantly. Though, higher concentrations of TiO 2 NPs influenced osteoblast cell cycle without interfering in their ability to differentiate and mineralize. For higher concentration of TiO 2 NPs, collagen deposition and pro-inflammatory cytokine, chemokine and growth factor secretion (involved in osteolysis and bone homeostasis) increased. These results raise the possible use of this model in nanotoxicological studies of osseointegrated devices and demonstrate a possible therapeutic potential of this TiO 2 NPs to prevent or reverse bone resorption.
BackgroundQuantitative real time polymerase chain reaction (qPCR) is an extremely powerful technique for monitoring gene expression. The quantity of the messenger ribonucleic acids (mRNA) of interest should be normalized using a reference gene, in order to avoid unreliable results originated by the obtained RNA quality and quantity, manipulation errors and inhibitory contaminants. A reference gene is any gene that is stably and consistently expressed under the conditions being studied. Completely false data can be generated if a reference gene is not chosen adequately.ResultsIn the present study, we compared expression levels of five putative reference genes (HPRT1, ACTB, GAPDH, RPL13A and B2M) in primary cultures of four different human cells: mesenchymal stromal cells obtained from bone marrow, adipose tissue or umbilical cord Whartońs Jelly, and dermal fibroblasts, under different expansion and differentiation conditions. We observed that reference genes are not the same for different cells under the same culture conditions.ConclusionMost stable reference genes under our experimental conditions were: RPL13A for adipose tissue- and Whartońs Jelly-derived mesenchymal stromal cells, and HPRT1 for bone marrow-derived mesenchymal stromal cells and dermal fibroblasts. ACTB was the most unstable gene when evaluating adipose tissue- and Whartońs Jelly-derived mesenchymal stromal cells, whilst GAPDH and B2M were the most unstable genes for bone marrow-derived mesenchymal stromal cells and dermal fibroblasts, respectively.
Studies indicate that tooth crown diameters are clinical markers for sex differentiation. Therefore, the aim of this study was to assess the degree of sexual dimorphism in different teeth. Maximum mesiodistal (MD) and buccolingual (BL) dimensions of 2400 permanent teeth from 100 pretreatment orthodontic dental study casts and clinical records (50 males and 50 females) from the Department of Pediatric Dentistry and Orthodontics, Federal University of Rio de Janeiro, Brazil, were examined. Comparison of the MD and BL dimensions between males and females was performed using the Student's t test with alpha 0.05, effect size, and discriminant function analysis. Comparisons in MD and BL widths between sexes demonstrated that the combined mean in the female group presented reduction when compared with the male group, except for the BL dimension of tooth 26. In regard to the MD dimensions, statistically significant differences were observed in various dental groups. The greatest sexual dimorphism was observed in the left mandibular canine (p<0.001) with effect size over 0.8 (0.94), which characterizes large effect. In BL dimension, numerous teeth demonstrated statistical differences between the sexes. Our findings reinforced the magnitude of sexual dimorphism in tooth size, and, in addition, highlighted the differences in specific dental groups.
An acid phosphatase (EC 3.1.3.2) has been identified and purified from castor bean (Ricinus communis L., IAC‐80) seed through sulphopropyl (SP)‐Sephadex, diethylaminoethyl (DEAE)‐Sephadex, Sephacryl S‐200, and Concanavalin A‐Sepharose chromatography. The enzyme was purified 2 000‐fold to homogeneity, with a final specific activity of 3.8 μkat mg−1 protein. The purified enzyme revealed a single diffuse band with phosphatase activity on nondenaturing polyacrylamide gel electrophoresis, at pH 8.3. The relative molecular mass, determined by high‐performance liquid chromatography (HPLC), was found to be 60 kDa. The acid phosphatase had a pH optimum of 5.5 and an akpparent Km value for p‐nitrophenylphosphate of 0.52 mM. The enzyme‐catalyzed reaction was inhibited by inorganic phosphate, fluoride, vanadate, molybdate, p‐chloromercuribenzoate (pCMB), Cu2+ and Zn2+. The strong inhibition by pCMB, Cu2+ and vanadate suggests the presence of sulfhydryl groups essential for catalysis. The castor bean enzyme also recognized tyrosine‐phosphate and inorganic pyrophosphate (KPPi) as substrate. The highest specificity constant (Vmax/Km) was observed with KPPi, making it a potential physiological substrate.
A low molecular weight bovine kidney acid phosphatase, electrophoretically homogeneous and with a relative molecular mass of 17.8 kDa, was used in this work. Among the various substrates tested, FMN was found to be the most effective, at pH 7.0. Distinct activation energy values were obtained for p‐nitrophenyl phosphate‐ (45.44 kJ mol‐1) and flavin mononucleotide‐ (28.60 kJ mol‐1) hydrolysis reactions. The FMN hydrolysis was strongly inhibited by Cu2+ and pCMB, but activated by guanosine. Pyridoxal‐phosphate and vanadate were competitive inhibitors for the FMN‐dependent reaction.
Chocolate bars and chocolate cookies are foodstuffs highly appreciated by children. The possibility of having fluorine (F) among their components, associated with an excessive consumption, may make them decisive contributors to the total daily F intake. Thus, they could participate in the establishment of dental fluorosis. The aim of this study was to analyze the fluorine concentration [F] of the chocolates bars (CB) Baton, Confeti, Garoto Ball, Kinder Ovo, M&M's, Milkybar, Nescau, Nescau Ball, Surpresa, Surpresa Bichos, Tortuguita; and of the chocolate cookies (CC) Danyt's, Hipopó, Nescau, Passatempo, Pokémon, Sítio do Pica-Pau Amarelo and Trakinas. Samples were purchased in Bauru, São Paulo, Brazil. Three grams of each product were previously ashed at 525°C (CB and cookies fillings) and at 550°C (cookies dough), during 4 hours. Fluorine was separated from the ash by hexamethyldisiloxane (HMDS)-facilitated diffusion. Fluorine analysis was carried out with the specific electrode. Mean [F]s ± SD and amplitude (unit mg/g) were: CB = 0.30 ± 0.45 (0.07 -1.60, n = 12) and CC = 1.08 ± 2.64 (0.04 -7.10, n = 7). It was concluded that some of the analyzed foods may be important contributors to the total daily F intake. As for the product that had the highest [F] (Danyt's), when only 3 units are consumed just once a day, they may supply up to 40% of the maximum recommended daily F intake (0.07 mg/kg body weight) for a 2-year-old child (12 kg). The [F] in these products should be informed on their labels. DESCRIPTORS: Chocolate; Fluorine; Fluorosis, dental; Child. RESUMO:Chocolates em barra e bolachas de chocolate são guloseimas altamente apreciadas pelas crianças. A possibilidade de conter flúor (F) em seus componentes, associada a seu excessivo consumo podem torná-los contribuintes decisivos para a ingestão diária total de F. Assim, eles poderiam participar no estabelecimento da fluorose dental. O objetivo deste estudo foi analisar a concentração de flúor [F] dos chocolates em barra (CB) Baton, Confeti, Garoto Ball, Kinder Ovo, M&M's, Milkybar, Nescau, Nescau Ball, Surpresa, Surpresa Bichos, Tortuguita; e das bolachas de chocolate (CC) Danyt's, Hipopó, Nescau, Passatempo, Pokémon, Sítio do Pica-Pau Amarelo e Trakinas. Os produtos foram adquiridos em Bauru, São Paulo, Brasil. Três gramas de cada produto foram previamente calcinadas a uma temperatura de 525°C (CB e recheio das bolachas), e de 550°C (massas das bolachas), durante 4 horas. O F foi separado das cinzas por difusão facilitada por hexametildisiloxano (HMDS). As análises de F foram feitas com o eletrodo específico. As [F]s médias ± DP e amplitude (mg/g) foram: CB = 0,30 ± 0,45 (0,07 -1,60, n = 12) e CC = 1,08 ± 2,64 (0,04 -7,10, n = 7). Concluiu-se que alguns dos alimentos analisados podem ser importantes contribuintes para a ingestão diária total de F. No caso do produto que apresentou a maior [F] (Danyt's), quando apenas 3 unidades são consumidas uma única vez ao dia, elas podem fornecer mais de 40% da ingestão diária máxima de F recomendada (0,07 m...
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