Crystallinity of TiO2 nanotubes and its effects on fibroblast viability, adhesion, and proliferation.

Titanium and titanium alloys are widely used as a biomaterial due to their mechanical strength, corrosion resistance, low elastic modulus, and excellent biocompatibility. TiO2 nanotubes have excellent bioactivity, stimulating the adhesion, proliferation of fibroblasts and adipose-derived stem cells, production of alkaline phosphatase by osteoblasts, platelets activation, growth of neural cells and adhesion, spreading, growth, and differentiation of rat bone marrow mesenchymal stem cells. In this study, we investigated the functionality of fibroblast on titania nanotube layers annealed at different temperatures. The titania nanotube layer was fabricated by potentiostatic anodization of titanium, then annealed at 300, 530, and 630 °C for 5 h. The resulting nanotube layer was characterized using SEM (Scanning Electron Microscopy), TF-XRD (Thin-film X-ray diffraction), and contact angle goniometry. Fibroblasts viability was determined by the CellTiter-Blue method and cytotoxicity by Lactate Dehydrogenase test, and the cell morphology was analyzed by scanning electron microscopy. Also, cell adherence, proliferation, and morphology were analyzed by fluorescence microscopy. The results indicate that the modification in nanotube crystallinity may provide a favorable surface fibroblast growth, especially on substrates annealed at 530 and 630 °C, indicating that these properties provide a favorable template for biomedical implants.

Effect of crystalline phases of titania nanotube arrays on adipose derived stem cell adhesion and proliferation.

The aim of this work was to evaluate the cellular response to titanium nanotube arrays with variable crystalline structure. Cytotoxicity, viability and the ability of the titania nanotube arrays to stimulate adhesion and proliferation of adipose derived stem cells (ADSCs) was evaluated. Titania nanotube arrays were fabricated by electrochemical anodization of titanium in diethyleneglycol/hydrofluoric acid electrolyte at 60 V for 6 h, then annealed at 300, 530 and 630 °C for 5 h. The nanotube arrays were characterized using scanning electron microscopy (SEM), contact angle goniometry, x-ray diffraction (XRD) and protein adsorption. ADSCs were cultured on titania nanotube arrays at a density of 1 × 104 cells/ml. The cells were allowed to adhere and to proliferate for 1, 4 and 7 days. Cell viability was characterized by the CellTiter-Blue® Cell Viability Assay; and cell morphology was characterized by SEM. Cell adhesion, proliferation and morphology were characterized using fluorescence microscopy by staining the cells with DAPI and rhodamine/phalloidin. The results from this study showed that the annealing at 300 and 530 °C formed anatase phase, and annealing at 630 °C formed anatase/rutile phase. The results indicated that the modification of the crystalline structure (i.e. anatase/rutile phase) of titania nanotube arrays influenced the ADSC adhesion and proliferation. Future studies are now directed towards evaluating differentiation of this cellular model in osteoblasts.

Antitumoral effects of Amblyomma sculptum Berlese saliva in neuroblastoma cell lines involve cytoskeletal deconstruction and cell cycle arrest / Efeito antitumoral da saliva do carrapato Amblyomma sculptum Berlese em células de neuroblastoma envolve desconstrução do citoesqueleto e parada do ciclo celular

Abstract The antitumor properties of ticks salivary gland extracts or recombinant proteins have been reported recently, but little is known about the antitumor properties of the secreted components of saliva. The goal of this study was to investigate the in vitro effect of the saliva of the hard tick Amblyomma sculptum on neuroblastoma cell lines. SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32, and CHLA-20 cells were susceptible to saliva, with 80% reduction in their viability compared to untreated controls, as demonstrated by the methylene blue assay. Further investigation using CHLA-20 revealed apoptosis, with approximately 30% of annexin-V positive cells, and G0/G1-phase accumulation (>60%) after treatment with saliva. Mitochondrial membrane potential (Δψm) was slightly, but significantly (p < 0.05), reduced and the actin cytoskeleton was disarranged, as indicated by fluorescent microscopy. The viability of human fibroblast (HFF-1 cells) used as a non-tumoral control decreased by approximately 40%. However, no alterations in cell cycle progression, morphology, and Δψm were observed in these cells. The present work provides new perspectives for the characterization of the molecules present in saliva and their antitumor properties.

Resumo As propriedades antitumorais de extratos de glândulas salivares de carrapatos ou proteínas recombinantes foram relatadas recentemente, mas pouco se sabe sobre as propriedades antitumorais dos componentes secretados da saliva. O objetivo deste estudo foi investigar o efeito in vitro da saliva bruta do carrapato duro Amblyomma sculptum sobre as linhagens celulares de neuroblastoma. Células SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32 e CHLA-20 foram suscetíveis à saliva, com redução de 80% na sua viabilidade em comparação com controles não tratados, como demonstrado pelo ensaio de Azul de Metileno. Investigações posteriores utilizando CHLA-20 revelaram apoptose, com aproximadamente 30% de células positivas para anexina-V, e G0/G1 (> 60%) após tratamento com saliva. O potencial de membrana mitocondrial (Δψm) foi reduzido significativamente (p <0,05), e o citoesqueleto de actina foi desestruturado, como indicado pela microscopia de fluorescência. A viabilidade do fibroblasto humano (células HFF-1), usado como controle não tumoral, diminuiu em aproximadamente 40%. No entanto, não foram observadas alterações na progressão do ciclo celular, morfologia e Δψm nestas células. O presente trabalho fornece novas perspectivas para a caracterização das moléculas presentes na saliva e suas propriedades antitumorais.

Antitumoral effects of Amblyomma sculptum Berlese saliva in neuroblastoma cell lines involve cytoskeletal deconstruction and cell cycle arrest.

The antitumor properties of ticks salivary gland extracts or recombinant proteins have been reported recently, but little is known about the antitumor properties of the secreted components of saliva. The goal of this study was to investigate the in vitro effect of the saliva of the hard tick Amblyomma sculptum on neuroblastoma cell lines. SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32, and CHLA-20 cells were susceptible to saliva, with 80% reduction in their viability compared to untreated controls, as demonstrated by the methylene blue assay. Further investigation using CHLA-20 revealed apoptosis, with approximately 30% of annexin-V positive cells, and G0/G1-phase accumulation (>60%) after treatment with saliva. Mitochondrial membrane potential (Δψm) was slightly, but significantly (p < 0.05), reduced and the actin cytoskeleton was disarranged, as indicated by fluorescent microscopy. The viability of human fibroblast (HFF-1 cells) used as a non-tumoral control decreased by approximately 40%. However, no alterations in cell cycle progression, morphology, and Δψm were observed in these cells. The present work provides new perspectives for the characterization of the molecules present in saliva and their antitumor properties.

Effects of titania nanotube surfaces on osteogenic differentiation of human adipose-derived stem cells.

The surface of an implant is important for successful osseointegration and long-term stability as it can aid in cell migration and proliferation, cell differentiation and allow extracellular matrix production. Earlier studies have shown that nanostructuring the surface of titanium can enhance mesenchymal stem cell (MSC) migration, proliferation, and differentiation. Although many studies have evaluated MSC response on nanostructured surfaces, there are only a few studies that have explored the response of adipose-derived stem cells (ADSC) on titania nanotube surfaces. Because ADSC exhibit great potential in regenerative medicine and have already proven effective in developing new treatments, this study aims to further understand how ADSC interact with titania nanotube surfaces. The results of this study indicate that titania nanotube surfaces enhance ADSC proliferation and differentiation that is also dependent on the size of nanotubes. Additionally, the favorable response of ADSC on nanotube surfaces suggests a potential application in orthopedic tissue regeneration.

Antitumoral effects of Amblyomma sculptum Berlese saliva in neuroblastoma cell lines involve cytoskeletal deconstruction and cell cycle arrest

The antitumor properties of ticks salivary gland extracts or recombinant proteins have been reported recently, but little is known about the antitumor properties of the secreted components of saliva. The goal of this study was to investigate the in vitro effect of the saliva of the hard tick Amblyomma sculptum on neuroblastoma cell lines. SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32, and CHLA-20 cells were susceptible to saliva, with 80% reduction in their viability compared to untreated controls, as demonstrated by the methylene blue assay. Further investigation using CHLA-20 revealed apoptosis, with approximately 30% of annexin-V positive cells, and G0/G1-phase accumulation (>60%) after treatment with saliva. Mitochondrial membrane potential (??m) was slightly, but significantly (p < 0.05), reduced and the actin cytoskeleton was disarranged, as indicated by fluorescent microscopy. The viability of human fibroblast (HFF-1 cells) used as a non-tumoral control decreased by approximately 40%. However, no alterations in cell cycle progression, morphology, and ??m were observed in these cells. The present work provides new perspectives for the characterization of the molecules present in saliva and their antitumor properties.

Antitumoral effects of Amblyomma sculptum Berlese saliva in neuroblastoma cell lines involve cytoskeletal deconstruction and cell cycle arrest

The antitumor properties of ticks salivary gland extracts or recombinant proteins have been reported recently, but little is known about the antitumor properties of the secreted components of saliva. The goal of this study was to investigate the in vitro effect of the saliva of the hard tick Amblyomma sculptum on neuroblastoma cell lines. SK-N-SK, SH-SY5Y, Be(2)-M17, IMR-32, and CHLA-20 cells were susceptible to saliva, with 80% reduction in their viability compared to untreated controls, as demonstrated by the methylene blue assay. Further investigation using CHLA-20 revealed apoptosis, with approximately 30% of annexin-V positive cells, and G0/G1-phase accumulation (>60%) after treatment with saliva. Mitochondrial membrane potential (??m) was slightly, but significantly (p < 0.05), reduced and the actin cytoskeleton was disarranged, as indicated by fluorescent microscopy. The viability of human fibroblast (HFF-1 cells) used as a non-tumoral control decreased by approximately 40%. However, no alterations in cell cycle progression, morphology, and ??m were observed in these cells. The present work provides new perspectives for the characterization of the molecules present in saliva and their antitumor properties.

Effects of calcium and phosphorus incorporation on the properties and bioactivity of TiO2 nanotubes.

In this study, we investigate the formation of calcium and phosphorus-doped TiO2 nanotubes, produced by potentiostatic anodization of Ti in viscous electrolyte-containing HF and Ca/P ions. Characterization of the produced oxide layer was conducted using scanning electron microscopy, glancing-angle X-ray diffraction, X-ray photoelectron spectroscopy, contact angle, and protein adsorption measurements. Adipose-derived stem cells were used to study material cytotoxicity, cell viability and proliferation, and cell morphology and growth. To evaluate the adipose-derived stem-cell differentiation, we investigated the expression of osteocalcin and osteopontin by cells as well as calcium mineralization. Results show that it was possible to produce a superhydrophilic titanium oxide nanotube layer with incorporation of Ca and P ions. The presence of Ca and P in the oxide layer not only improved the cell adhesion and proliferation but also stimulated the production of key marker proteins indicating differentiation of cells.

In-vitro cell adhesion and proliferation of adipose derived stem cell on hydroxyapatite composite surfaces.

The goal of this work was to enhance the mechanical strength and fracture toughness of brittle hydroxyapatite (HAP) by reinforcing it with nanocomposites such as graphene oxide (GO), carbon nanotubes (CNT) and Titania. The goal was also to evaluate the cytotoxicity and the cellular adhesion/proliferation of these composites. The composites were characterized for their crystallinity, functionality, morphology and mechanical properties. Altering the composition by adding 1wt% GO and CNT significantly altered the wettability, hardness and roughness. Further, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FITR) and X-ray photoelectron spectroscopy (XPS) results confirm the crystal structure, bulk chemical composition and surface elemental composition respectively of the composites. The bulk hardness of HAP with CNT was significantly higher than that of HAP. The wettability of HAP with GO was significantly lower than that of HAP with GO and Titania. Adipose Derived Stem Cells (ADSCs) were used for this study to evaluate cytotoxicity and viability. HAP with CNT and HAP with CNT and Titania were found to be least cytotoxic compared to other composites as evaluated by Lactate Dehydrogenase (LDH) assay and alamarBlue assay. ADSC adhesion and proliferation was investigated after 1, 4 and 7days of culture using fluorescence microscopy. All the composites nurtured ADSC adhesion and proliferation, however, distinct morphological changes were observed by using Scanning Electron Microscopy (SEM). Overall, these composites have the potential to be used as bone graft substitutes.