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Antisense nucleic acid drugs are susceptible to nuclease degradation, rapid renal clearance, and short circulatory half-life. In this work, we introduce a modular-based recombinant human albumin-oligonucleotide (rHA-cODN) biomolecular assembly that allows incorporation of a chemically stabilized therapeutic gapmer antisense oligonucleotide (ASO) and FcRn-driven endothelial cellular recycling. A phosphodiester ODN linker (cODN) was conjugated to recombinant human albumin (rHA) using maleimide chemistry, after which a complementary gapmer ASO, targeting ADAMTS5 involved in osteoarthritis pathogenesis, was annealed. The rHA-cODN/ASO biomolecular assembly production, fluorescence labeling, and purity were confirmed using polyacrylamide gel electrophoresis. ASO release was triggered by DNase-mediated degradation of the linker strand, reaching 40% in serum after 72 h, with complete release observed following 30 min of incubation with DNase. Cellular internalization and trafficking of the biomolecular assembly using confocal microscopy in C28/I2 cells showed higher uptake and endosomal localization by increasing incubation time from 4 to 24 h. FcRn-mediated cellular recycling of the assembly was demonstrated in FcRn-expressing human microvascular endothelial cells. ADAMTS5 in vitro silencing efficiency reached 40%, which was comparable to free gapmer after 72 h incubation with human osteoarthritis patients' chondrocytes. This work introduces a versatile biomolecular modular-based "Plug-and-Play" platform potentially applicable for albumin-mediated half-life extension for a range of different types of ODN therapeutics.
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Oligonucleotídeos , Osteoartrite , Humanos , Oligonucleotídeos/química , Células Endoteliais/metabolismo , Albuminas , Oligonucleotídeos Antissenso/química , Albumina Sérica Humana/metabolismo , DesoxirribonucleasesRESUMO
OBJECTIVE: To evaluate the effect of grafting with strontium (Sr)-loaded deproteinized bovine bone (DBB) on bone healing in calvarial critical size defects (CSD) in rats. MATERIAL AND METHODS: Two circular bone defects (5 mm in diameter) were created in the calvaria of 42 rats. One of the defects, randomly chosen, was grafted with (a) DBB, (b) DBB loaded with 19.6 µg/g of Sr (DBB/Sr1), or (c) DBB loaded with 98.1 µg/g of Sr (DBB/Sr2). The other defect was left empty as negative control. Groups of seven animals from each of the groups were euthanized 15 and 60 days post-op. Bone healing in the CSD was evaluated by micro-CT and histology/histomorphometry and immunohistochemistry. RESULTS: DBB/Sr2-grafted sites showed statistically significantly shorter radiographic residual defect length compared with DBB/Sr1- and DBB-grafted sites, and with empty controls at 60 days. Further, the amount of new bone formation in the DBB/Sr1- and DBB/Sr2-grafted sites was significantly higher compared with that in the DBB-grafted sites at 60 days. A larger number of DBB/Sr1- and DBB/Sr2-grafted sites presented with no- or only limited to mild inflammation, compared with the DBB-grafted sites, especially at 60 days. Higher expression of osteocalcin was observed in DBB/Sr1- and DBB/Sr2-grafted sites as compared to DBB-grafted sites. CONCLUSION: Grafting with Sr-loaded DBB enhanced bone formation in CSD in rats, when compared with grafting with non-loaded DBB. CLINICAL RELEVANCE: Grafting with Sr-loaded DBB may enhance bone formation in bone defects.
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Regeneração Óssea , Substitutos Ósseos , Estrôncio , Animais , Bovinos , Feminino , Osteogênese , Ratos , CrânioRESUMO
BACKGROUND AND OBJECTIVE: Strontium (Sr) enhances osteogenic differentiation of certain multipotent cells. Periodontal ligament cells (PDLCs) are known to be multipotent, and Sr might be useful in periodontal bone tissue engineering. This study investigates the effect of high concentration of Sr on the proliferation and osteogenic behavior of PDLCs in vitro. MATERIAL AND METHODS: Primary human PDLCs were cultured in MEM + 10% FBS without (Ctrl) or with Sr in four diverse concentrations: Sr1, 11.3 × 10-3 mg/L, human serum physiological level; Sr2, 13 mg/L, typical human serum level after strontium ranelate treatment; Sr3, 130 mg/L, and Sr4, 360 mg/L. The spreading area (2, 4, 6, 24 hours), proliferation rate (1, 3, 7 days), osteogenic behavior (alkaline phosphatase - ALP activity, 7 and 14 days; expression of osteogenic genes, ALP, Runt-related transcription factor 2 - RUNX2, osteopontin - OPN, osteocalcin - OCN, and osteoprotegerin -OPG, 1, 3, 7, 14, 21 days), and formation of mineralized nodules (14 and 21 days) of the PDLCs were assessed. Data were compared group- and period-wise using ANOVA tests. RESULTS: Periodontal ligament cells cultured with Sr4 showed increased spreading area (after 4 hours), proliferation rate (from 3 days), and OCN and OPN (from 7 days) gene expression as compared to Ctrl, Sr1, Sr2, and Sr3. Sr4 also led to lower ALP activity (from 7 days), ALP (from 3 days), and RUNX2 (at 7 and 14 days) gene expression, together with more evident formation of mineralized nodules, compared to Ctrl, Sr1, Sr2, and Sr3. CONCLUSION: Periodontal ligament cells responded to Sr4 with increased cellular proliferation and osteogenic behavior in vitro.
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Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Ligamento Periodontal/citologia , Estrôncio/farmacologia , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Relação Dose-Resposta a Droga , Feminino , Expressão Gênica/efeitos dos fármacos , Humanos , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogênese/genética , Osteopontina/genética , Osteopontina/metabolismo , Osteoprotegerina/genética , Osteoprotegerina/metabolismo , Estimulação Química , Engenharia Tecidual , Adulto JovemRESUMO
Biofunctional surface patterns capable of resisting nonspecific bioadsorption while retaining bioactivity play crucial roles in the advancement of life science and biomedical technologies. The currently available functional surface coatings suffer from a high level of nonspecific surface adsorption of proteins under biologically challenging conditions, leading to a loss of activity in functional moieties over time. In this study, the recently discovered facile method of temperature-induced polyelectrolyte (TIP) grafting has been used to graft two biofunctional variants (biotin and nitrilotriacetic acid, NTA) of poly(l-lysine)-grafted PEG (PLL-g-PEG) onto a titanium surface. A significant increase in the polymer adsorption was observed from the TIP-grafted surfaces assembled at 80 °C, compared to the polymer surfaces assembled at ambient temperature (20 °C). These functional PLL-g-PEG surfaces were subsequently incubated in whole human blood continuously for up to 7 days, and the TIP-grafted surfaces achieved close-to-zero nonspecific protein adsorption, as confirmed by ultrasensitive time-of-flight secondary ion mass spectrometry (ToF-SIMS). To test the maintenance of the bioactivity of the biotin and NTA moieties, submicrometer-scale mono- (biotin) and bi- (biotin/NTA) functional surface chemical patterns were fabricated via two-step TIP grafting using colloidal lithography (CL), preincubated in blood for up to 7 days and sequentially exposed to streptavidin and Ni(2+)-histidine-tagged calmodulin. The fluorescence microscopy studies revealed that the PLL-g-PEG-NTA and -biotin surfaces grafted from the TIP method were still capable of recognizing the corresponding affinity proteins for up to 1 and 7 days of preincubation in blood, respectively. These results highlight the bioresistant robustness realized by the facile TIP grafting method, which in turn preserves the activities of biofunctional moieties over a prolonged period in whole blood.
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Polímeros/química , Adsorção , Humanos , Polietilenoglicóis/química , Polilisina/análogos & derivados , Polilisina/química , Propriedades de Superfície , Titânio/químicaRESUMO
OBJECTIVES: To investigate the impact of a Ti-Sr-O technology, applied to either a turned surface or an SLA surface, on the mechanical robustness of osseointegration, benchmarked against the SLActive surface. MATERIAL AND METHODS: Ti discs (6.25-mm-diameter and 2-mm-thick) with three different surfaces were inserted on the proximal-anterior part of the tibial plateau of adult Swedish loop rabbits: (I) turned surface modified with Ti-Sr-O (turned + Ti-Sr-O), (II) SLA surface modified with Ti-Sr-O (SLA + Ti-Sr-O), and (III) SLActive surface (SLActive). Following a healing period of 2 weeks and 4 weeks, the pull-out (PO) force needed to detach the discs from the bone was assessed, as a surrogate of osseointegration. RESULTS: The SLActive surface exhibited statistically significant higher median PO forces, compared with the SLA + Ti-Sr-O surfaces at both 2- and 4 weeks post-op (p > .05). In this study, no single turned + Ti-Sr-O surface disk was integrated. CONCLUSIONS: The tested Ti-Sr-O technology failed to enhance osseointegration; however, this finding may be related to the inappropriateness of the rabbit tibia plateau model for assessing third-generation implant surface technologies, due to the limited diffusion and clearance at the disk-bone interface.
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Implantes Dentários , Osseointegração , Óxidos , Titânio , Animais , Coelhos , Tíbia/cirurgia , Propriedades de Superfície , EstrôncioRESUMO
The article mentioned in the title of this comment paper reports on an investigation of the organic binder presence and distribution on stone wool fibres with surface sensitive techniques (X-ray photoelectron spectroscopy (XPS), QUASES XPS modelling, time-of-flight secondary ion mass spectrometry (ToF-SIMS) mapping) and attempts to correlate the results with fibre performance in in vitro acellular biosolubility tests. However, the study has assumptions, hypothesis and results that do not take into account the recognised science and regulations on biopersistence of stone wool fibres, limitations of the utilized surface sensitive techniques and modelling approach and it contains a contradiction with biosolubility experiments. In this comment article, we discuss these points, propose improved QUASES XPS modelling and present recent ToF-SIMS mapping results that reflect biosolubility behaviour of the stone wool fibres.
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The immobilization of enzymes on solid supports is an important challenge in biotechnology and biomedicine. In contrast to other methods, enzyme deposition in polymer brushes offers the benefit of high protein loading that preserves enzymatic activity in part due to the hydrated 3D environment that is available within the brush structure. The authors equipped planar and colloidal silica surfaces with poly(2-(diethylamino)ethyl methacrylate)-based brushes to immobilize Thermoplasma acidophilum histidine ammonia lyase, and analyzed the amount and activity of the immobilized enzyme. The poly(2-(diethylamino)ethyl methacrylate) brushes are attached to the solid silica supports either via a "grafting-to" or a "grafting-from" method. It is found that the grafting-from method results in higher amounts of deposited polymer and, consequently, higher amounts of Thermoplasma acidophilum histidine ammonia lyase. All polymer brush-modified surfaces show preserved catalytic activity of the deposited Thermoplasma acidophilum histidine ammonia lyase. However, immobilizing the enzyme in polymer brushes using the grafting-from method resulted in twice the enzymatic activity from the grafting-to approach, illustrating a successful enzyme deposition on a solid support.
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Histidina Amônia-Liase , Polímeros , Polímeros/química , Metacrilatos/química , Dióxido de SilícioRESUMO
Biochemical and biomechanical signals regulate stem cell function in the niche environments in vivo. Current in vitro culture of mouse embryonic stem cells (mESC) uses laminin (LN-511) to provide mimetic biochemical signaling (LN-521 for human systems) to maintain stemness. Alternative approaches propose topographical cues to provide biomechanical cues, however combined biochemical and topographic cues may better mimic the in vivo environment, but are largely unexplored for in vitro stem cell expansion. In this study, we directly compare in vitro signals from LN-511 and/or topographic cues to maintain stemness, using systematically-varied submicron pillar patterns or flat surfaces with or without preadsorbed LN-511. The adhesion of cells, colony formation, expression of the pluripotency marker,octamer-binding transcription factor 4 (Oct4), and transcriptome profiling were characterized. We observed that either biochemical or topographic signals could maintain stemness of mESCs in feeder-free conditions, indicated by high-level Oct4 and gene profiling by RNAseq. The combination of LN-511 with nanotopography reduced colony growth, while maintaining stemness markers, shifted the cellular phenotype indicating that the integration of biochemical and topographic signals is antagonistic. Overall, significantly faster (up to 2.5 times) colony growth was observed at nanotopographies without LN-511, suggesting for improved ESC expansion.
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Células-Tronco Embrionárias , Células-Tronco Embrionárias Murinas , Animais , Camundongos , Humanos , Células Cultivadas , Ligantes , Fator 3 de Transcrição de Octâmero/genética , Fator 3 de Transcrição de Octâmero/metabolismo , Fenótipo , Diferenciação Celular/fisiologiaRESUMO
The development of cost-effective methodologies for the precise nanometer-scale positioning of biomolecules permits the low-cost production of various biofunctional devices for a range of biomedical and nanotechnological applications. By combining colloidal lithography and the mussel-inspired multifunctional polydopamine coating, we present a novel parallel benchtop method that allows rapid nanoscale patterning of proteins without the need for electrically powered equipment in the fabrication process. The PDA-immobilized binary nanopattern consisting of BSA surrounded by PLL-g-PEG is fabricated over a large area, and the integrity of the pattern is confirmed using AFM and FM.
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Dopamina/química , Nanotecnologia/métodos , Polietilenoglicóis/química , Polilisina/análogos & derivados , Impressão/métodos , Proteínas/química , Coloides , Microscopia de Força Atômica , Polilisina/química , Análise Serial de Proteínas , Propriedades de SuperfícieRESUMO
We report a facile method of generating ultradense poly(l-lysine)-graft-poly(ethylene glycol) (PLL-g-PEG) surface by using high temperature alone, which in turn provides dramatic improvement in resisting nonspecific bioadsorption. X-ray photoelectron spectroscopy (XPS) revealed that the surface graft density increased ~4 times higher on the surface prepared at 80 °C compared to 20 °C. The studies from small-angle X-ray scattering (SAXS) and the effect of varying ionic strength during/post assemblies at 20 and 80 °C indicated that the "cloud point grafting effect" is not the cause for obtaining high density grafting. Stringent long-term bioresistance tests have been conducted and the temperature-induced PLL-g-PEG surfaces have achieved (1) zero mammalian cell adsorption/migration for up to 36 days and (2) extremely close-to-zero protein adsorptions have been observed even after 36 days in 10% serum media and 24 h in whole blood within the ultrasensitive detection limit of time-of-flight secondary ion mass spectrometry (ToF-SIMS).
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Materiais Revestidos Biocompatíveis/química , Polietilenoglicóis/química , Polilisina/análogos & derivados , Transplantes , Adsorção , Animais , Sangue , Células Cultivadas , Humanos , Concentração Osmolar , Espectroscopia Fotoeletrônica , Polilisina/sangue , Polilisina/química , Propriedades de Superfície , TemperaturaRESUMO
The characterization of roughness at the nanoscale by the means of atomic force microscopy (AFM) was performed on high aspect ratio glancing angle deposited titanium thin films. With the use of scanning electron microscopy as well as x-ray photoelectron spectroscopy, it was shown that the AFM measurements gave rise to incorrect roughness values for the films consisting of the highest aspect ratio structures. By correcting for this experimental artefact, the difference between the saturated roughness value of a film grown with conventional physical vapour deposition and films grown with a glancing angle of deposition was shown to behave as a power law function of the deposition angle, with a saturated roughness exponent of κ = 7.1 ± 0.2. This power law scaling was confirmed by three-dimensional molecular dynamics simulations of glancing angle deposition, where the saturated roughness exponent was calculated to κ = 6.7 ± 0.4.
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Membranas Artificiais , Modelos Químicos , Modelos Moleculares , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Titânio/química , Simulação por Computador , Substâncias Macromoleculares/química , Teste de Materiais , Conformação Molecular , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
It is well known that strontium (Sr) has a significant effect on peri-implant bone healing when administered systemically. Due to the risk of adverse effects of such treatments, new routes focusing on the local, sustained release of Sr from bone-implant contact surfaces have been explored, with success in in vivo experiments. However, the increase of Sr concentrations in the peri-implant bone has not been described in depth yet. Here, we show that a local, sustained Sr release from Ti-Sr-O physical vapor deposition (PVD) coatings by magnetron sputter coating increases the Sr/Ca ratio close to the implant in a rabbit model and that the Sr/Ca background level is reached approximately 500 µm from the implant.
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Osseointegração , Estrôncio , Animais , Cálcio/farmacologia , Materiais Revestidos Biocompatíveis/farmacologia , Coelhos , Estrôncio/farmacologia , Propriedades de Superfície , Titânio/farmacologiaRESUMO
Nano- and micromotors are self-navigating particles that gain locomotion using fuel from the environment or external power sources to outperform Brownian motion. Herein, motors that make use of surface polymerization of hydroxyethylmethylacrylate to gain locomotion are reported, synthetically mimicking microorganisms' way of propulsion. These motors have enhanced Brownian motion with effective diffusion coefficients up to â¼0.5 µm2 s-1 when mesoporous Janus particles are used. Finally, indication of swarming is observed when high numbers of motors homogenously coated with atom-transfer radical polymerization initiators are used, while high-density Janus motors lost their ability to exhibit enhanced Brownian motion. This report illustrates an alternative route to self-propelled particles, employing a polymerization process that has the potential to be applied for various purposes benefiting from the tool box of modern polymer chemistry.
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Polydopamine (PDA) is the final oxidation product of dopamine or other catecholamines. Since the first reports of PDA coatings starting around 2007, these coatings have been widely studied as a versatile and inexpensive one-step coating option for biomaterial functionalization. The coating attach to a wide range of materials and can subsequently be modified with biomolecules or nanoparticles. However, as a strong candidate for biomaterial research and even clinical use, it is important to unravel the changes in physico-chemical properties and the cell-PDA interaction as a function of heat sterilization procedures and shelf storage periods. Four groups were examined in this study: titanium (Ti), PDA-coated Ti samples and PDA-coated Ti samples either stored for up to two weeks at room temperature or heated at 121 °C for 24 h, respectively. We used X-ray Photoelectron Spectroscopy (XPS), Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) and Water contact angle (WCA) to characterize chemical composition and surface properties of the groups. Cell adhesion and proliferation was examined by three different cell types: human primary dermal fibroblasts (hDF), human epidermal keratinocytes (HaCaTs) and a murine preosteoblastic cell line (MC3T3-E1), respectively. Cells were cultured on PDA coated samples for 4 h, 3 days and 5 days. Both thermal treatment of PDA at 121â for 24 h and storage of the samples for 2 weeks increased the amount of quinone groups at the surface and decreased the amount of primary amine groups as detected by XPS and ToF-SIMS. Even though these surface reactions increased the WCA of the PDA coating, we found that the post-treatments increased cell proliferation for both hDFs, HaCaTs and MC3T3-E1 s as compared to pristine PDA. This emphasizes the importance of post-treatment and shelf-time for PDA coatings.
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Materiais Biocompatíveis , Indóis , Animais , Adesão Celular , Humanos , Indóis/farmacologia , Queratinócitos , Camundongos , PolímerosRESUMO
Obejective: To investigate the effect of increasing Strontium (Sr) concentrations on the growth and osteogenic behavior of human bone marrow stromal cells (BMSCs) from mesenchymal (i.e., fibula) and ectomesenchymal (i.e., mandible) embryonic origins. Materials and methods: Fibula and mandible BMSCs were cultured in media without (Ctrl) or with Sr in four diverse concentrations: Sr1, 11.3 × 10-3 mg/L, human seric physiological level; Sr2, 13 mg/L, human seric level after strontium ranelate treatment; Sr3, 130 mg/L, and Sr4, 360 mg/L. Proliferation rate (1, 3, and 7 days), osteogenic behavior (alkaline phosphatase [ALP] activity, 7 and 14 days; expression of osteogenic genes (ALP, osteopontin, and osteocalcin at 7, 14, and 21 days), and formation of mineralized nodules (14 and 21 days) of the BMSCs were assessed. Data was compared group- and period-wise using analysis of variance tests. Results: Fibula and mandible BMSCs cultured with Sr4 showed increased proliferation rate, and osteocalcin and osteopontin gene expression together with more evident formation of mineralized nodules, compared all other Sr concentrations. For both cell populations, Sr4 led to lower ALP activity, and ALP gene expression, compared with the other Sr concentrations. Conclusion: BMSCs from mesenchymal (i.e., fibula) and ectomesenchymal (i.e., mandible) embryonic origins showed increased cellular proliferation and osteogenic behavior when cultured with Sr4, in vitro.
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Calcificação Fisiológica , Células-Tronco Mesenquimais/citologia , Mesoderma/citologia , Osteogênese , Estrôncio/farmacologia , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Diferenciação Celular , Células Cultivadas , Humanos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Mesoderma/efeitos dos fármacos , Mesoderma/metabolismo , Pessoa de Meia-Idade , Osteocalcina/genética , Osteocalcina/metabolismo , Osteopontina/genética , Osteopontina/metabolismoRESUMO
A cell's ability to remodel adsorbed protein layers on surfaces is influenced by the nature of the protein layer itself. Remodelling is often required to accomplish cellular adhesion and extracellular matrix formation which forms the basis for cell spreading, increased adhesion and expression of different phenotypes. The adhesion of NIH3T3 (EGFP) fibroblasts to serum protein (albumin or fibronectin) precoated tantalum (Ta) and oxidised polystyrene (PS(ox)) surfaces was examined using the quartz crystal microbalance with dissipation (QCM-D) monitoring and fluorescence microscopy. The cells were either untreated or treated with cycloheximide to examine the contribution of endogenous protein production during cell adhesion to the QCM-D response over a period of 2h. Following adsorption of albumin onto Ta and PS(ox) there was no difference detected between the response to seeding untreated and cycloheximide treated cells. The QCM-D was able to detect differences in the untreated cellular responses to fibronectin versus serum precoated Ta and PS(ox) substrates, while cycloheximide treatment of the cells produced the same QCM-D response for fibronectin and serum precoatings on each of the materials. This confirmed that the process of matrix remodelling by the cells is dependent on the underlying substrate and the preadsorbed proteins and that the QCM-D response is dominated by changes in the underlying protein layer. Changes in dissipation correspond to the development of the actin cytoskeleton as visualised by actin staining.
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Materiais Revestidos Biocompatíveis/química , Cicloeximida/farmacologia , Matriz Extracelular/metabolismo , Inibidores da Síntese de Proteínas/farmacologia , Quartzo , Adsorção , Animais , Bovinos , Adesão Celular/efeitos dos fármacos , Técnicas de Cultura de Células , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Fibronectinas/química , Camundongos , Microscopia de Fluorescência , Células NIH 3T3 , Oxirredução , Poliestirenos/química , Soro/química , Soroalbumina Bovina/química , Ovinos , Especificidade por Substrato , Tantálio/química , Fatores de TempoRESUMO
The complex mechanisms of protein adsorption at the solid-liquid interface is of great importance in many research areas, including protein purification, biocompatibility of medical implants, biosensing, and biofouling. The protein adsorption process depends crucially on both the nanoscale chemistry and topography of the interface. Here, we investigate the adsorption of the cell-binding protein fibronectin on flat and nanometer scale rough tantalum oxide surfaces using ellipsometry and quartz crystal microbalance with dissipation (QCM-D). On the flat tantalum oxide surfaces, the interfacial protein spreading causes an increase in the rigidity and a decrease in the thickness of the adsorbed fibronectin layer with decreasing bulk protein concentration. For the tantalum oxide surfaces with well-controlled, stochastic nanometer scale roughness, similar concentration effects are observed for the rigidity of the fibronectin layer and saturated fibronectin uptake. However, we find that the nanorough tantalum oxide surfaces promote additional protein conformational changes, an effect especially apparent from the QCM-D signals, interpreted as an additional stiffening of the formed fibronectin layers.
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Fibronectinas/química , Nanoestruturas/química , Tantálio/química , Adsorção , Anticorpos/imunologia , Cristalização , Fibronectinas/imunologia , Fibronectinas/ultraestrutura , Humanos , Microscopia de Força Atômica , Nanoestruturas/ultraestrutura , Óxidos/química , Propriedades de SuperfícieRESUMO
The adsorption of fibronectin on gold, Ti-, and Ta-oxide surfaces is investigated by means of the quartz crystal microbalance with dissipation (QCM-D) technique. The surface chemistry (gold, Ti-, and Ta-oxide) is found to influence the frequency shift observed during adsorption of the fibronectin layer with the magnitude being Delta f Au>Delta f Ti-oxide approximately Delta f Ta-oxide. Corresponding variations in the dissipation change normalised to frequency change (Delta D/Delta f) for the layer are observed. The QCM-D data are further analyzed by the random sequential adsorption (RSA) model, and adsorption rate parameter ka and footprint (a) determined, which supported the trend seen in the Delta f and Delta D/Delta f values. The value of ka found by the RSA modelling of the QCM-D resonance frequency data is found to match the ratio between the mass measured by QCM-D and the mass reported by optical techniques in literature. We conclude that comparison of the adsorption rate parameter (ka) obtained by RSA modelling of the QCM-D data with ka values obtained from RSA modelling of data obtained using optical techniques can be a route to determine the degree of hydration of the adsorbed protein layer.
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Fibronectinas/química , Ouro/química , Óxidos/química , Tantálio/química , Titânio/química , Adsorção , Humanos , Quartzo/química , Propriedades de SuperfícieRESUMO
PURPOSE: Studies have shown that strontium-doped medical applications benefit bone metabolism leading to improved bone healing and osseointegration. Based on this knowledge, the aim of the study was to evaluate the performance of an implant surface, functionalized by a physical vapor deposition (PVD) coating (Ti-Sr-O), designed to yield predictable release of strontium. The Ti-Sr-O functionalized surface is compared to a routinely used, commercially available surface (SLActive™) with respect to bone-to-implant contact (BIC%) and new bone formation (BF%) in two defined regions of interest (ROI-I and ROI-II, respectively). MATERIALS AND METHODS: Ti-Sr-O functionalized, SLActive, and Grade 4 titanium implants were inserted in the femoral condyle of adult male New Zealand White rabbits. The PVD magnetron-sputtered Ti-Sr-O surface coating was characterized using scanning electron microscopy (SEM) for morphology and coating thickness. Strontium release and mechanical stability of the coating, under simulated insertion conditions, were evaluated. Furthermore, histomorphometrical BIC and BF were carried out 2 weeks after insertion. RESULTS: Histomorphometry revealed increased bone formation of Ti-Sr-O with significant differences compared to SLActive and Grade 4 titanium in both regions of interest, ROI-I and ROI-II, at 0-250 µm and 250-500 µm distance from the implant surfaces. Analogous results of bone-to-implant contact were observed for the two modified surfaces. CONCLUSION: The results show that a nanopatterned Ti-Sr-O functionalized titanium surface, with sustained release of strontium, increases peri-implant bone volume and could potentially contribute to enhancement of bone anchorage of osseointegrated implants.
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Interface Osso-Implante/fisiologia , Próteses e Implantes , Estrôncio/farmacocinética , Animais , Osso e Ossos/fisiologia , Fêmur/fisiologia , Fêmur/cirurgia , Masculino , Microscopia Eletrônica de Varredura , Osseointegração/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia , Coelhos , Estrôncio/química , Propriedades de Superfície , Titânio/químicaRESUMO
Understanding the behavior of chondrocytes in contact with artificial culture surfaces is becoming increasingly important in attaining appropriate ex vivo culture conditions of chondrocytes in cartilage regeneration. Chondrocyte transplantation-based cartilage repair requires efficiently expanded chondrocytes, and the culture surface plays an important role in guiding the behavior of the cell. Micro- and nano-engineered surfaces make it possible to modulate cell behavior. We hypothesized that the combined influence of topography, substrate, and surface chemistry may affect the chondrocyte culturing in terms of proliferation and phenotypic means. Human chondrocytes were cultured on polystyrene fabricated microstructures, flat polydimethylsiloxane (PDMS), or polystyrene treated with fibronectin or oxygen plasma and cultured for 1, 4, 7, and 10 days. The behavior of chondrocytes was evaluated by proliferation, viability, chondrogenic gene expression, and cell morphology. Contrary to our hypothesis, microstructures in polystyrene did not significantly influence the behavior of chondrocytes neither under normoxic- nor hypoxic conditions. However, changes in the substrate stiffness and surface chemistry were found to influence cell viability, gene expression, and morphology of human chondrocytes. Oxygen plasma treatment was the most important parameter followed by the softer substrate type PDMS. The findings indicate the culture of human chondrocytes on softer substratum and surface activation by oxygen plasma may prevent dedifferentiation and may improve chondrocyte transplantation-based cartilage repair. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2805-2816, 2018.