RESUMO
The Franz cells permeation assay has been performed for over 25 years. However, the advent of nanotechnology created a whole new world, especially with regard to topical products. In this new global scenario an increasing number of nanostructure-based delivery systems (NDSs) have emerged and a global warning relating to the safety of these NDSs is arising. This work studied the efficacy of the Franz cells assay, comparing it with the radiolabeling biodistribution test. For this purpose a formulation of sunscreen based on an NDS was developed and characterized. The results demonstrated both that the NDS did not present in vitro cytotoxicity and that the radiolabeling biodistribution test is more precise for the evaluation of NDS cosmetics than the Franz cells assay, since it detected the permeation of the NDS at a picogram order. Due to this fact, and considering all the concerns related to NDSs and nanoparticles in general, more precise methods must be used in order to guarantee the safe use of these new classes of products.
Assuntos
Portadores de Fármacos/química , Nanopartículas/química , Absorção Cutânea , Protetores Solares/administração & dosagem , Protetores Solares/farmacocinética , Animais , Linhagem Celular , Portadores de Fármacos/toxicidade , Emulsões/química , Emulsões/toxicidade , Haplorrinos , Camundongos , Nanopartículas/toxicidade , Ratos Wistar , Pele/metabolismo , Protetores Solares/toxicidade , Distribuição TecidualRESUMO
Cell cultures are often used to study bone mineralization; however, not all systems achieve a bone-like matrix formation. In this study, the mineralized matrix assembled in F-OST osteoblast cultures was analyzed, with the aim of establishing a novel model for bone mineralization. The ultrastructure of the cultures was investigated using scanning electron microscopy, atomic force microscopy, and transmission electron microscopy (TEM). The mineral phase was characterized using conventional and high-resolution TEM, energy-dispersive X-ray spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, and solid-state (31)P and (1)H nuclear magnetic resonance. F-OST osteoblast cultures presented a clear nodular mineralization pattern. The chief features of the mineralizing nodules were globular accretions ranging from about 100 nm to 1.5 µm in diameter, loaded with needle-shaped crystallites. Accretions seemed to bud from the cell membrane, increase in size, and coalesce into larger ones. Arrays of loosely packed, randomly oriented collagen fibrils were seen along with the accretions. Mineralized fibrils were often observed, sometimes in close association with accretions. The mineral phase was characterized as a poorly crystalline hydroxyapatite. The Ca/P atomic ratio was 1.49 ± 0.06. The presence of OH was evident. The lattice parameters were a = 9.435 Å and c = 6.860 Å. The average crystallite size was 20 nm long and 10 nm wide. Carbonate substitutions were seen in phosphate and OH sites. Water was also found within the apatitic core. In conclusion, F-OST osteoblast cultures produce a bone-like matrix and may provide a good model for bone mineralization studies.
Assuntos
Matriz Óssea/ultraestrutura , Osteoblastos/citologia , Animais , Matriz Óssea/química , Calcificação Fisiológica , Células Cultivadas , Durapatita/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Microscopia de Força Atômica , Microscopia Eletrônica de Transmissão , Osteoblastos/metabolismo , Osteoblastos/ultraestrutura , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
One of the main problems that remain in the implant industry is poor osseointegration due to bioinertness of implants. In order to promote bioactivity, calcium (Ca), phosphorus (P) and strontium (Sr) were incorporated into a TiO2 porous layer produced by micro-arc oxidation. Ca and P as bioactive elements are already well reported in the literature, however, the knowledge of the effect of Sr is still limited. In the present work, the effect of various amounts of Sr was evaluated and the morphology, chemical composition and crystal structure of the oxide layer were investigated. Furthermore, in vitro studies were carried out using human osteoblast-like cells. The oxide layer formed showed a triplex structure, where higher incorporation of Sr increased Ca/P ratio, amount of rutile and promoted the formation of SrTiO3 compound. Biological tests revealed that lower concentrations of Sr did not compromise initial cell adhesion neither viability and interestingly improved mineralization. However, higher concentration of Sr (and consequent higher amount of rutile) showed to induce collagen secretion but with compromised mineralization, possibly due to a delayed mineralization process or induced precipitation of deficient hydroxyapatite. Ca-P-TiO2 porous layer with less concentration of Sr seems to be an ideal candidate for bone implants.
Assuntos
Materiais Revestidos Biocompatíveis , Estrôncio , Humanos , Osseointegração , Propriedades de Superfície , TitânioRESUMO
Precambrian cellular remains frequently have simple morphologies, micrometric dimensions and are poorly preserved, imposing severe analytical and interpretational challenges, especially for irrefutable attestations of biogenicity. The 1.88 Ga Gunflint biota is a Precambrian microfossil assemblage with different types and qualities of preservation across its numerous geological localities and provides important insights into the Proterozoic biosphere and taphonomic processes. Here we use synchrotron-based ptychographic X-ray computed tomography to investigate well-preserved carbonaceous microfossils from the Schreiber Beach locality as well as poorly-preserved, iron-replaced fossil filaments from the Mink Mountain locality, Gunflint Formation. 3D nanoscale imaging with contrast based on electron density allowed us to assess the morphology and carbonaceous composition of different specimens and identify the minerals associated with their preservation based on retrieved mass densities. In the Mink Mountain filaments, the identification of mature kerogen and maghemite rather than the ubiquitously described hematite indicates an influence from biogenic organics on the local maturation of iron oxides through diagenesis. This non-destructive 3D approach to microfossil composition at the nanoscale within their geological context represents a powerful approach to assess the taphonomy and biogenicity of challenging or poorly preserved traces of early microbial life, and may be applied effectively to extraterrestrial samples returned from upcoming space missions.
RESUMO
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 (TiO2 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 TiO2 NPs during 72 h did not change their viability significantly. Though, higher concentrations of TiO2 NPs influenced osteoblast cell cycle without interfering in their ability to differentiate and mineralize. For higher concentration of TiO2 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 TiO2 NPs to prevent or reverse bone resorption.
Assuntos
Nanopartículas/toxicidade , Osteoblastos/citologia , Esferoides Celulares/citologia , Esferoides Celulares/efeitos dos fármacos , Titânio/farmacologia , Titânio/toxicidade , Ciclo Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Citocinas/metabolismo , Relação Dose-Resposta a Droga , Homeostase/efeitos dos fármacos , Humanos , Minerais/metabolismo , Esferoides Celulares/metabolismo , Titânio/químicaRESUMO
The use of nanoparticles (NPs) in the healthcare market is growing exponentially, due to their unique physicochemical properties. Titanium dioxide nanoparticles (TiO2 NPs) are used in the formulation of sunscreens, due to their photoprotective capacity, but interactions of these particles with skin cells on the nanoscale are still unexplored. In the present study we aimed to determine whether the initial nano-biological interactions, namely the formation of a nano-bio-complex (other than the protein corona), can predict rutile internalization and intracellular trafficking in primary human fibroblasts and keratinocytes. Results showed no significant effect of NPs on fibroblast and keratinocyte viability, but cell proliferation was possibly compromised due to nano-bio-interactions. The bio-complex formation is dependent upon the chemistry of the biological media and NPs' physicochemical properties, facilitating NP internalization and triggering autophagy in both cell types. For the first time, we observed that the intracellular traffic of NPs is different when comparing the two skin cell models, and we detected NPs within multivesicular bodies (MVBs) of keratinocytes. These structures grant selected input of molecules involved in the biogenesis of exosomes, responsible for cell communication and, potentially, structural equilibrium in human tissues. Nanoparticle-mediated alterations of exosome quality, quantity and function can be another major source of nanotoxicity.
RESUMO
Dentistry and orthopedics are undergoing a revolution in order to provide more reliable, comfortable and long-lasting implants to patients. Titanium (Ti) and titanium alloys have been used in dental implants and total hip arthroplasty due to their excellent biocompatibility. However, Ti-based implants in human body suffer surface degradation (corrosion and wear) resulting in the release of metallic ions and solid wear debris (mainly titanium dioxide) leading to peri-implant inflammatory reactions. Unfortunately, our current understanding of the biological interactions with titanium dioxide nanoparticles is still very limited. Taking this into consideration, this study focuses on the internalization of titanium dioxide nanoparticles on primary bone cells, exploring the events occurring at the nano-bio interface. For the first time, we report the selective binding of calcium (Ca), phosphorous (P) and proteins from cell culture medium to anatase nanoparticles that are extremely important for nanoparticle internalization and bone cells survival. In the intricate biological environment, anatase nanoparticles form bio-complexes (mixture of proteins and ions) which act as a kind of 'Trojan-horse' internalization by cells. Furthermore, anatase nanoparticles-induced modifications on cell behavior (viability and internalization) could be understand in detail. The results presented in this report can inspire new strategies for the use of titanium dioxide nanoparticles in several regeneration therapies.
Assuntos
Endocitose , Nanopartículas Metálicas/química , Osteoblastos/metabolismo , Titânio/metabolismo , Cálcio/metabolismo , Sobrevivência Celular , Células Cultivadas , Citoplasma/metabolismo , Citoplasma/ultraestrutura , Humanos , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica , Osteoblastos/citologia , Osteoblastos/ultraestrutura , Tamanho da Partícula , Fósforo/metabolismo , Ligação Proteica , Titânio/química , Difração de Raios XRESUMO
OBJECTIVE: To describe the development and initial psychometric properties of a new outcome measure for health behaviors that delay or prevent secondary impairments associated with spinal cord injury (SCI). DESIGN: Persons with SCI were surveyed during routine annual physical evaluations. SETTING: Veterans Affairs Medical Center Spinal Cord Injury Unit, which specializes in primary care for persons with SCI. PARTICIPANTS: Forty-nine persons with SCI, aged 19-73 years, 1-50 years post-SCI. MAIN OUTCOME MEASURE: The newly developed Spinal Cord Injury Lifestyle Scale (SCILS). RESULTS: Internal consistency is high (alpha = 0.81). Correlations between clinicians' ratings of participants' health behavior and the new SCILS provide preliminary support for construct validity. CONCLUSIONS: The SCILS is a brief, self-report measure of health-related behavior in persons with SCI. It is a promising new outcome measure to evaluate the effectiveness of clinical and educational efforts for health maintenance and prevention of secondary impairments associated with SCI.
Assuntos
Comportamentos Relacionados com a Saúde , Traumatismos da Medula Espinal/psicologia , Adolescente , Adulto , Idoso , Feminino , Humanos , Estilo de Vida , Masculino , Pessoa de Meia-Idade , Psicometria , Qualidade de Vida , Reprodutibilidade dos Testes , Traumatismos da Medula Espinal/complicações , Traumatismos da Medula Espinal/reabilitação , Traumatismos da Medula Espinal/terapia , Resultado do TratamentoAssuntos
Encefalite por Arbovirus , Encefalomielite Equina , Adolescente , Adulto , Idoso , Doenças dos Animais/microbiologia , Animais , Animais Domésticos , Formação de Anticorpos , Vetores Artrópodes , Aves , Encefalopatias/microbiologia , Criança , Pré-Escolar , Clima , Culicidae , Técnicas de Cultura , Demografia , Reservatórios de Doenças , Vetores de Doenças , Vírus da Encefalite/isolamento & purificação , Encefalomielite Equina/sangue , Encefalomielite Equina/diagnóstico , Encefalomielite Equina/microbiologia , Encefalomielite Equina/mortalidade , Exsudatos e Transudatos/microbiologia , Feminino , Geografia , Testes de Inibição da Hemaglutinação , Humanos , Lactente , Masculino , Mamíferos , Pessoa de Meia-Idade , Testes de Neutralização , População Rural , VenezuelaAssuntos
Neoplasias Pancreáticas/diagnóstico por imagem , Portografia , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeAssuntos
Síndrome de Down/sangue , Fosfolipídeos/sangue , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-IdadeRESUMO
La vitamina C incluye dos formas activas, ácido ascórbico (AA) y ácido dehidroascórbico (ADHA), que funcionan como un sistema redox interconvertible. El nivel de vitamina C en vegetales está condicionado por factores de orden climático y agronómico, por el manejo poscosecha y las condiciones de almacenamiento. Por tratarse de un nutriente extremadamente lábil, se producen pérdidas o deterioro durante su procesamiento. En los vegetales recién cosechados la vitamina se encuentra mayormente como AA, pero gradualmente pasa a ADHA, su forma oxidada. El ADHA es muy inestable y al escindirse su anillo lactónico se pierde la actividad vitamínica. A los efectos de determinar el nivel de vitamina C en jugos de limón fresco y procesado, tal cual lo adquiere el consumidor, se analizó el contenido de vitamina C y de sus formas biológicamente activas, en muestras tomadas al azar en supermercados y negocios minoristas. Los resultados obtenidos demuestran que en los jugos analizados, los tiempos de poscosecha, las condiciones de almacenamiento y el procesado influyen notablemente sobre el contenido de la vitamina
Assuntos
Ácido Ascórbico , Bebidas/análise , Citrus/uso terapêutico , Ácido Ascórbico/análise , Cromatografia Líquida de Alta Pressão/tendências , Oxidantes/antagonistas & inibidoresRESUMO
Estudaram-se os niveis de lipideos totais, colesterol e triglicerideos no soro e na fracao alfa-lipoproteina (HDL), em 11 pacientes hipotireoideos, 39 hipertiroideos e 21 individuos normais. Em termos percentuais, os pacientes hipertireoideos manifestaram maior percentagem de colesterol e triglicerideos na fracao alfa-lipoproteina. A fracao colesterol total HDL/colesterol foi mais elevada nos hipotireoideos. Nao houve diferenca estatisticamente significante para os valores absolutos dos lipideos da fracao alfa-lipoproteina entre hipo e hipertiroideos