RESUMO
The application of algae to the most meaningful fields of our life, such as food, environment and energy, finds a further confirmation in the extension of this application to cultural heritage protection. In this letter, we present the results of a preliminary study testing how a polysaccharide extracted from algal matrix can restore degraded paper giving back it mechanical strength and chemical structure.
Assuntos
Fenômenos Mecânicos/efeitos dos fármacos , Papel , Extratos Vegetais/química , Cultura , Recuperação e Remediação Ambiental , Humanos , Papel/normas , Extratos Vegetais/farmacologia , Polissacarídeos/química , Polissacarídeos/farmacologia , Rodófitas/químicaRESUMO
This study aimed to evaluate the effects of different synthetic and natural-derived root canal irrigants (6% sodium hypochlorite [NaOCl], 6% calcium hypochlorite [Ca(OCl)2] and 6.5% grape seed extract [GSE]) on dentin mechanical properties (flexural strength, ultimate tensile strength [UTS] and fracture resistance). Rectangular-shaped beams and hourglass-shaped sections obtained from mid-coronal and root dentin were treated with 6% NaOCl, 6% Ca(OCl)2 or 6.5% GSE for 30 min. The irrigant solutions were replaced every 5 min. Then, the dentin specimens were rinsed with distilled water (DW) followed by incubation with 17% EDTA for 1 min, and thoroughly rinsed with DW again. Specimens from the control group were tested without prior irrigation. After treatment with the irrigants, dentin beams were used to assess the flexural strength (n = 10) while UTS was evaluated using the root dentin hourglass-shaped sections (n = 10). Similarly, roots with 1 mm of dentinal wall thickness were obtained from human teeth and treated with the same irrigant solutions (n = 10). A compressive loading was applied to the coronal surfaces of roots until fracture. The values of each mechanical test were statistically analyzed individually by one-way ANOVA followed by Tukey HSD test (P < 0.05). NaOCl significantly reduced the mechanical properties of dentin in all mechanical tests (P < 0.05) and no statistical difference was found among Ca(OCl)2, GSE and control group (P > 0.05). It can be concluded that Ca(OCl)2 and GSE may be alternative irrigant solutions, since they do not negatively affect the dentin mechanical properties.
Assuntos
Compostos de Cálcio/farmacologia , Dentina/efeitos dos fármacos , Extrato de Sementes de Uva/farmacologia , Irrigantes do Canal Radicular/farmacologia , Hipoclorito de Sódio/farmacologia , Análise do Estresse Dentário/métodos , Humanos , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Estresse Mecânico , Resistência à Tração/efeitos dos fármacos , Dente/efeitos dos fármacos , Dente/fisiologia , Fraturas dos Dentes/prevenção & controleRESUMO
Treatment with lipid-lowering drugs, statins, is common all over the world. Lately, the occurrence of spontaneous tendon ruptures or tendinosis have suggested a negative influence of statins upon tendon tissue. But how statins might influence tendons is not clear. In the present study, we investigated the effect of statin treatment on mechanical strength, cell proliferation, collagen content and gene expression pattern in a tendon-like tissue made from human tenocytes in vitro. Human tendon fibroblasts were grown in a 3D tissue culture model (tendon constructs), and treated with either simvastatin or atorvastatin, low or high dose, respectively, for up to seven days. After seven days of treatment, mechanical testing of the constructs was performed. Collagen content and cell proliferation were also determined. mRNA levels of several target genes were measured after one or seven days. The maximum force and stiffness were reduced by both statins after 7 days (p<0.05), while the cross sectional area was unaffected. Further, the collagen content was reduced by atorvastatin (p = 0.01) and the cell proliferation rate was decreased by both types of statins (p<0.05). Statin treatment also introduced increased mRNA levels of MMP-1, MMP-3, MMP-13, TIMP-1 and decreased levels of collagen type 1 and 3. In conclusion, statin treatment appears to have a negative effect on tendon matrix quality as seen by a reduced strength of the tendon constructs. Further, activated catabolic changes in the gene expression pattern and a reduced collagen content indicated a disturbed balance in matrix production of tendon due to statin administration.
Assuntos
Atorvastatina/farmacologia , Metabolismo Energético/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Inibidores de Hidroximetilglutaril-CoA Redutases/farmacologia , Fenômenos Mecânicos/efeitos dos fármacos , Sinvastatina/farmacologia , Tendões/efeitos dos fármacos , Tendões/fisiologia , Adolescente , Adulto , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Colágeno/genética , Colágeno/metabolismo , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Humanos , Metaloproteinases da Matriz/genética , Metaloproteinases da Matriz/metabolismo , Traumatismos dos Tendões/genética , Traumatismos dos Tendões/metabolismo , Traumatismos dos Tendões/fisiopatologia , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Adulto JovemRESUMO
BACKGROUND: Arterial function is regulated by estrogen, but no consistent pattern of arterial mechanical remodeling in response to depleted estrogen levels is available. OBJECTIVE: To examine long-term effects of ovariectomy (OVX) on the mechanical properties, morphology, and histological structure of the carotid artery in middle-aged rats and a potentially protective effect of Sideritis euboea extract (SID), commonly consumed as "mountain tea". METHODS: 10-month-old female Wistar rats were allocated into control (sham-operated), OVX, OVX+SID, and OVX+MALT (maltodextrin; excipient used for dilution of SID) groups. They were sacrificed after 6 months and their carotid arteries were submitted to inflation/extension tests and to dimensional and histological evaluation. RESULTS: Remodeling in OVX rats was characterized by a decreased in situ axial extension ratio, along with increased opening angle, thickness, and area of the vessel wall and of its medial layer, but unchanged lumen diameter. Compositional changes involved increased elastin/collagen densities. Characterization by the "four-fiber" microstructure-motivated model revealed similar in situ biaxial response of carotid arteries in OVX and control rats. CONCLUSIONS: Carotid artery remodeling in OVX rats was largely consistent with hypertensive remodeling, despite the minor arterial pressure changes found, and was not altered by administration of SID, despite previous evidence of its osteo-protective effect.
Assuntos
Artérias Carótidas/efeitos dos fármacos , Artérias Carótidas/fisiologia , Fenômenos Mecânicos/efeitos dos fármacos , Ovariectomia/efeitos adversos , Extratos Vegetais/farmacologia , Sideritis/química , Animais , Fenômenos Biomecânicos/efeitos dos fármacos , Peso Corporal/efeitos dos fármacos , Artérias Carótidas/anatomia & histologia , Feminino , Hemodinâmica/efeitos dos fármacos , Tamanho do Órgão/efeitos dos fármacos , Extratos Vegetais/administração & dosagem , Ratos , Ratos Wistar , Útero/efeitos dos fármacos , Útero/crescimento & desenvolvimentoRESUMO
In this study, water-soluble, one-step highly reduced and functionalized graphene oxide was prepared via a facile, environment-friendly method by using tea polyphenol (TP), which acted as both reducing agent and stabilizer. The product obtained, that is, tea polyphenol-reduced graphene oxide (TPG), was used as a reinforcing building block for the modification of a mechanically weak chitosan (CS), TPG/CS. The morphology and physicochemical and mechanical properties of the composite were examined by various characterizations. The tensile strength and elastic modulus of CS were greatly improved by TPG, as compared to the findings for GO incorporation. Additionally, to our knowledge, this study is an in-depth analysis of the osteoblast functions of CS/TPG, including aspects such as cell cytotoxicity, proliferation, and expression of ossification genes, alkaline phosphatase (ALP), and Runt-related transcription factor (Runx2), which showed advantages in favorably modulating cellular activity. It was concluded that TPG/CS showed a higher elastic modulus, better hydrophilicity, and excellent biocompatibility than the pristine chitosan for promoting the proliferation and differentiation of osteoblasts, as well as for accelerating the expression of ALP and Runx2 (as shown by reverse transcription polymerase chain reaction (RT-PCR)). These results may provide new prospects for the use of TPG in the modification of biomaterials and for broadening the application of TPG in biological fields.
Assuntos
Materiais Biocompatíveis/farmacologia , Quitosana/química , Grafite/química , Fenômenos Mecânicos/efeitos dos fármacos , Polifenóis/farmacologia , Chá/química , Animais , Regulação da Expressão Gênica/efeitos dos fármacos , Microscopia de Força Atômica , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/ultraestrutura , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Espectroscopia de Infravermelho com Transformada de Fourier , Análise Espectral Raman , Propriedades de Superfície , Difração de Raios XRESUMO
Bioceramic samples with osteogenic properties, suitable for use in the regeneration of hard tissue, were synthesized. The materials consisting of α-tricalcium phosphate (αTCP) and also αTCP doped with either 1.5 wt.% or 3.0 wt.% of dicalcium silicate (C2S) in the system Dicalcium Silicate-Tricalcium Phosphate (C2S-TCP) were obtained by solid state reaction. All materials were composed of a single phase, αTCP in the case of a pure material, or solid solution of C2S in αTCP (αTCPss) for the doped αTCP. Viability, proliferation and in vitro osteoinductive capacity were investigated by seeding, adult mesenchymal stem cells of human origin (ahMSCs) which were CD73(+), CD90(+), CD105(+), CD34(-) and CD45(-) onto the 3 substrates for 30 days. Results show a non-cytotoxic effect after applying an indirect apoptosis test (Annexin V/7-AAD staining), so ahMSCs adhered, spread, proliferated and produced extracellular matrix (Heparan-sulfate proteoglycan (HS) and osteopontin (OP)) on all the ceramics studied. Finally, the cells lost the cluster differentiation marker expression CD73, CD90 y CD105 characteristic of ahMSCs and they showed an osteoblastic phenotype (Alkaline phosphatase activity (ALP), Osteocalcin production (OC), Collagen type I expression (Col-I), and production of mineralization nodules on the extracellular matrix). These observations were more evident in the αTCP ceramic doped with 1.5 wt.% C2S, indicating osteoblastic differentiation as a result of the increased concentration of solid solution of C2S in αTCP (αTCPss). Overall, these results suggest that the ceramics studied are cytocompatible and they are able to induce osteoblastic differentiation of undifferentiated ahMSCs.
Assuntos
Células-Tronco Adultas/citologia , Compostos de Cálcio/farmacologia , Fosfatos de Cálcio/farmacologia , Diferenciação Celular/efeitos dos fármacos , Cerâmica/farmacologia , Células-Tronco Mesenquimais/citologia , Silicatos/farmacologia , Adulto , Células-Tronco Adultas/efeitos dos fármacos , Células-Tronco Adultas/enzimologia , Células-Tronco Adultas/ultraestrutura , Fosfatase Alcalina/metabolismo , Apoptose/efeitos dos fármacos , Cálcio/análise , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Fenômenos Químicos/efeitos dos fármacos , Meios de Cultura/química , Humanos , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/enzimologia , Células-Tronco Mesenquimais/ultraestrutura , Osteocalcina/metabolismo , Fósforo/análise , Silício/análiseRESUMO
Bone morphogenetic protein-2 (BMP2) is among the most popular anabolic agents and substantially increase bone volume related to enhanced osteoblast differentiation. Here we demonstrate a remarkable deterioration in the nanomechanical properties of mineralized tissue induced from osteoblasts solely by the function of BMP2. Mineralized tissue of primary osteoblasts cultured with BMP2 shows molecular features of both bone and cartilage, but depletion of lysyl oxidase family members leads to poor nanomechanical properties of the mineralized tissue. Lysyl oxidase like-2 supplementation reinforces the inferior mineralized tissue induced from osteoblasts by BMP2 through intermolecular cross-linking of type II or type X collagen-rich extracellular matrix. This may also mimic a consolidation of bone fracture gaps, despite the fact that the distribution of the bone properties in such microenvironments has been poorly elucidated. These findings confirm the importance of testing newly induced bone down to the microscale and nanoscale in bone tissue engineering. FROM THE CLINICAL EDITOR: Bone morphogenetic protein-2 is known to substantially increase bone volume related to enhanced osteoblast differentiation; however, this team of investigators report a remarkable deterioration in the nanomechanical properties of mineralized tissue induced from osteoblasts solely by the function of BMP2.
Assuntos
Aminoácido Oxirredutases/farmacologia , Proteína Morfogenética Óssea 2/farmacologia , Fenômenos Mecânicos/efeitos dos fármacos , Nanopartículas/química , Osteogênese/efeitos dos fármacos , Fator de Crescimento Transformador beta/farmacologia , Animais , Calcificação Fisiológica/efeitos dos fármacos , Células Cultivadas , Perfilação da Expressão Gênica , Masculino , Camundongos , Análise de Sequência com Séries de Oligonucleotídeos , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Proteínas Recombinantes/farmacologia , Análise Espectral RamanRESUMO
The present study investigates the potential use of non-catalyzed water-soluble blocked polyurethane prepolymer (PUP) as a bifunctional cross-linker for collagenous scaffolds. The effect of concentration (5, 10, 15 and 20%), time (4, 6, 12 and 24 h), medium volume (50, 100, 200 and 300%) and pH (7.4, 8.2, 9 and 10) over stability, microstructure and tensile mechanical behavior of acellular pericardial matrix was studied. The cross-linking index increased up to 81% while the denaturation temperature increased up to 12 °C after PUP crosslinking. PUP-treated scaffold resisted the collagenase degradation (0.167±0.14 mmol/g of liberated amine groups vs. 598±60 mmol/g for non-cross-linked matrix). The collagen fiber network was coated with PUP while viscoelastic properties were altered after cross-linking. The treatment of the pericardial scaffold with PUP allows (i) different densities of cross-linking depending of the process parameters and (ii) tensile properties similar to glutaraldehyde method.
Assuntos
Reagentes de Ligações Cruzadas/farmacologia , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Pericárdio/efeitos dos fármacos , Poliuretanos/farmacologia , Água/química , Animais , Cálcio/metabolismo , Varredura Diferencial de Calorimetria , Bovinos , Módulo de Elasticidade/efeitos dos fármacos , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/ultraestrutura , Glutaral/farmacologia , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Pericárdio/ultraestrutura , Fósforo/metabolismo , Estresse Mecânico , Temperatura , Resistência à Tração/efeitos dos fármacos , Fatores de TempoRESUMO
This study investigates the effect of insulin-like growth factor (IGF)-I on the development of anatomically-shaped alginate menisci seeded with meniscal fibrochondrocytes. To accomplish this, bovine meniscal fibrochondrocytes were seeded into 2% w/v alginate, crosslinked with calcium sulfate, and injected into anatomical molds derived from microcomputed tomography scans. The meniscal constructs were then cultured for up to 4 weeks with or without 100 ng/mL IGF-I supplemented in the media. Histological, immunohistological, biochemical, and mechanical analyses were performed to characterize tissue development, accumulation and localization of extracellular matrix, and mechanical properties. After 4 weeks of culture, IGF-I treatment significantly improved mechanical and biochemical properties, while maintaining DNA content, with a 26-fold increase in glycosaminoglycan (GAG) content and 10-fold increase in collagen content compared to 0-week controls, and a 3-fold increase in the equilibrium modulus at 2 weeks compared to controls. IGF-I-treated menisci had â¼60% of the GAG content of native tissue and the compressive equilibrium modulus matched native properties by 2 weeks of culture. Further, IGF-I-treated menisci developed a distinct surface layer similar to native tissue with elongated cells and collagen fibers aligned parallel to the surface, the presence of types I and II collagen, and accumulation of lubricin. This study demonstrates that IGF-I treatment can greatly increase the mechanical and biochemical properties of engineered tissues and aid in the development of a distinct surface zone similar to the superficial zone of native menisci.
Assuntos
Fator de Crescimento Insulin-Like I/farmacologia , Meniscos Tibiais/anatomia & histologia , Meniscos Tibiais/efeitos dos fármacos , Engenharia Tecidual/métodos , Alginatos/farmacologia , Animais , Bovinos , Colágeno Tipo I/metabolismo , Colágeno Tipo II/metabolismo , DNA/metabolismo , Ácido Glucurônico/farmacologia , Glicoproteínas/metabolismo , Glicosaminoglicanos/metabolismo , Ácidos Hexurônicos/farmacologia , Imuno-Histoquímica , Fenômenos Mecânicos/efeitos dos fármacos , Meniscos Tibiais/citologiaRESUMO
Factors influencing poly(3-hydroxybutyrate) P(3HB) production by Cupriavidus necator CCUG52238(T) utilizing oil palm frond (OPF) juice were clarified in this study. Effects of initial medium pH, agitation speed, and ammonium sulfate (NH(4))(2)SO(4) concentration on the production of P(3HB) were investigated in shake flasks experiments using OPF juice as the sole carbon source. The highest P(3HB) content was recorded at pH 7.0, agitation speed of 220 rpm, and (NH(4))(2)SO(4) concentration at 0.5 g/L. By culturing the wild-type strain of C. necator under the aforementioned conditions, the cell dry weight (CDW) and P(3HB) content obtained were 9.31 ± 0.13 g/L and 45 ± 1.5 wt.%, respectively. This accounted for 40% increment of P(3HB) content compared to the nonoptimized condition. In the meanwhile, the effect of dissolved oxygen tension (DOT) on P(3HB) production was investigated in a 2-L bioreactor. Highest CDW (11.37 g/L) and P(3HB) content (44 wt.%) were achieved when DOT level was set at 30%. P(3HB) produced from OPF juice had a tensile strength of 40 MPa and elongation at break of 8% demonstrated that P(3HB) produced from renewable and cheap carbon source is comparable to those produced from commercial substrate.
Assuntos
Cupriavidus necator/metabolismo , Hidroxibutiratos/metabolismo , Extratos Vegetais/metabolismo , Folhas de Planta/metabolismo , Óleos de Plantas/metabolismo , Poliésteres/metabolismo , Sulfato de Amônio/farmacologia , Biomassa , Reatores Biológicos/microbiologia , Cupriavidus necator/efeitos dos fármacos , Cupriavidus necator/crescimento & desenvolvimento , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Fenômenos Mecânicos/efeitos dos fármacos , Oxigênio/farmacologia , Óleo de Palmeira , Solubilidade , Temperatura , Fatores de TempoRESUMO
Tissue-engineered scaffolds may improve experimental outcomes in cardiac cell therapy by targeted delivery of stem cells and mechanically support an infarcted left ventricular (LV) wall. We transplanted cardiomyocyte-like cells (5×10(5)) with scaffolding via epicardial patching (cell patch, n=17) or a low-dose intramyocardial hydrogel (LD hydrogel, n=18), a high-dose (5×10(6)) intramyocardial hydrogel (HD hydrogel, n=18) or transplanting a serum-free medium control (control, n=13), a blank patch (n=14), and a blank gel (n=16) for targeted cardiomyoplasty in a myocardial infarcted rat model. LV real-time hemodynamics were assessed using a 1.9-F pressure-volume catheter 7 weeks after stem cell transplantation. All mode of scaffold transplantation protected diastolic function by preserving LV wall integrity that resulted in a lower end diastolic pressure-volume relationship (EDPVR) as compared to a control medium-injected group. Moreover, epicardial patching, but not hydrogel injection, reduced ventricular wall stress with a significantly better LV end diastolic pressure (EDP: 5.3±2.4 mmHg vs. 9.6±6.9 mmHg, p<0.05) as compared to control. Furthermore, epicardial patching additionally preserved systolic function by modulating negative remodeling through restricting dilatation of the LV chamber. In comparison to control, an improved ejection fraction in the cell patch group (80.1%±5.9% vs. 67.9%±3.2%, p<0.01) was corroborated by load-independent enhancement of the end systolic pressure-volume relationship (ESPVR: 0.88±0.61 mmHg/uL vs. 0.29±0.19 mmHg/uL, p<0.05) and preload recruitable stroke work (PRSW: 68.7±26.4 mmHg vs. 15.6±16.2 mmHg, p<0.05) in systolic function. Moreover, the cell patch group (14.2±1.7 cells/high-power field vs. 7.4±1.6 cells/high power field, p<0.05) was significantly better in myocardial retention of transplanted stem cells as compared to the LD hydrogel group. Collectively, myocardial transplantation of compliant scaffolding materials alone may physically improve wall mechanics, largely independent of stem cells. However, epicardially grafted cell patch conferred added systolic contractility by improving stem cell retention and cellular alignment leading to improved LV remodeling and geometric preservation postinfarction.
Assuntos
Hemodinâmica , Infarto do Miocárdio/fisiopatologia , Infarto do Miocárdio/terapia , Transplante de Células-Tronco , Células-Tronco/citologia , Alicerces Teciduais/química , Animais , Apoptose/efeitos dos fármacos , Materiais Biocompatíveis/farmacologia , Contagem de Células , Testes de Função Cardíaca , Ventrículos do Coração/efeitos dos fármacos , Ventrículos do Coração/fisiopatologia , Ventrículos do Coração/cirurgia , Hemodinâmica/efeitos dos fármacos , Humanos , Marcação In Situ das Extremidades Cortadas , Masculino , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Contração Miocárdica/efeitos dos fármacos , Infarto do Miocárdio/diagnóstico por imagem , Miócitos Cardíacos/citologia , Miócitos Cardíacos/transplante , Neovascularização Fisiológica/efeitos dos fármacos , Ratos , Ratos Wistar , UltrassonografiaRESUMO
Nickel-titanium (NiTi) shape memory alloys (SMAs) are commonly used in a range of biomedical applications. However, concerns exist regarding their use in certain biomedical scenarios due to the known toxicity of Ni and conflicting reports of NiTi corrosion resistance, particularly under dynamic loading. Titanium-niobium (TiNb) SMAs have recently been proposed as an alternative to NiTi SMAs due to the biocompatibility of both constituents, the ability of both Ti and Nb to form protective surface oxides, and their superior workability. However, several properties critical to the use of TiNb SMAs in biomedical applications have not been systematically explored in comparison with NiTi SMAs. These properties include cytocompatibility, corrosion resistance, and alterations in alloy surface composition in response to prolonged exposure to physiological solutions. Therefore, the goal of the present work was to comparatively investigate these aspects of NiTi (49.2 at.% Ti) and TiNb (26 at.% Nb) SMAs. The results from the current studies indicate that TiNb SMAs are less cytotoxic than NiTi SMAs, at least under static culture conditions. This increased TiNb cytocompatibility was correlated with reduced ion release as well as with increased corrosion resistance according to potentio-dynamic tests. Measurements of the surface composition of samples exposed to cell culture medium further supported the reduced ion release observed from TiNb relative to NiTi SMAs. Alloy composition depth profiles also suggested the formation of calcium phosphate deposits within the surface oxide layers of medium-exposed NiTi but not of TiNb. Collectively, the present results indicate that TiNb SMAs may be promising alternatives to NiTi for certain biomedical applications.
Assuntos
Ligas/toxicidade , Níquel/toxicidade , Titânio/toxicidade , Animais , Cálcio/análise , Morte Celular/efeitos dos fármacos , Corrosão , Elasticidade/efeitos dos fármacos , Íons , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Camundongos , Células NIH 3T3 , Níquel/análise , Nióbio/análise , Fósforo/análise , TemperaturaRESUMO
Poly(methylmethacrylate) (PMMA)-based cements containing magnetite (C-PMMA/Fe(3)O(4)) is useful in hyperthermia treatment for bone tumor. We have prepared C-PMMA/Fe(3)O(4) by incorporating Fe(3) O(4) powders of different diameters (means of 300, 35, and 11 nm) into the polymerization reaction of methyl methacrylate monomer to develop a new bone cement with high heating efficiencies in alternating current (AC) magnetic fields. Further, we have investigated the in vitro heating capability of the cements in different AC magnetic fields. The mechanical strength and biocompatibility of the resultant cements were also assessed. Their heat generation strongly depends on the magnetite nanoparticle sizes and applied magnetic fields. The cement containing Fe(3)O(4) with mean diameter around 35 nm exhibited the highest heating capability in AC magnetic fields of 120 and 300 Oe at 100 kHz while that with mean diameter around 11 nm exhibited optimum heating capability in AC magnetic fields of 40 Oe at 600 kHz. The incorporation of Fe(3)O(4) into cement-30 wt % of the total amount of cement-did not significantly change the compressive strength of cement, and the proliferation of rat fibroblast Rat-1 cells on cement discs was not inhibited. Our investigations are useful for designing new PMMA/Fe(3)O(4) bone cement with high heating efficiencies and biocompatibilities for bone tumor treatments.
Assuntos
Cimentos Ósseos/farmacologia , Neoplasias Ósseas/terapia , Hipertermia Induzida , Nanopartículas de Magnetita/uso terapêutico , Polimetil Metacrilato/farmacologia , Animais , Materiais Biocompatíveis/farmacologia , DNA/metabolismo , Eletricidade , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Temperatura Alta , Campos Magnéticos , Nanopartículas de Magnetita/ultraestrutura , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Ratos , Difração de Raios XRESUMO
The development of a new family of implantable bioinspired materials is a focal point of bone tissue engineering. Implant surfaces that better mimic the natural bone extracellular matrix, a naturally nano-composite tissue, can stimulate stem cell differentiation towards osteogenic lineages in the absence of specific chemical treatments. Herein we describe a bioactive composite nanofibrous scaffold, composed of poly-caprolactone (PCL) and nano-sized hydroxyapatite (HA) or beta-tricalcium phosphate (TCP), which was able to support the growth of human bone marrow mesenchymal stem cells (hMSCs) and guide their osteogenic differentiation at the same time. Morphological and physical/chemical investigations were carried out by scanning, transmission electron microscopy, Fourier-transform infrared (FTIR) spectroscopy, mechanical and wettability analysis. Upon culturing hMSCs on composite nanofibers, we found that the incorporation of either HA or TCP into the PCL nanofibers did not affect cell viability, meanwhile the presence of the mineral phase increases the activity of alkaline phosphatase (ALP), an early marker of bone formation, and mRNA expression levels of osteoblast-related genes, such as the Runt-related transcription factor 2 (Runx-2) and bone sialoprotein (BSP), in total absence of osteogenic supplements. These results suggest that both the nanofibrous structure and the chemical composition of the scaffolds play a role in regulating the osteogenic differentiation of hMSCs.
Assuntos
Materiais Biocompatíveis/farmacologia , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Alicerces Teciduais/química , Fosfatase Alcalina/metabolismo , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Fenômenos Mecânicos/efeitos dos fármacos , Células-Tronco Mesenquimais/enzimologia , Células-Tronco Mesenquimais/ultraestrutura , Nanofibras/ultraestrutura , Osteogênese/genética , Polímeros/química , Polímeros/farmacologia , Reação em Cadeia da Polimerase em Tempo Real , Espectroscopia de Infravermelho com Transformada de Fourier , Coloração e Rotulagem , Molhabilidade/efeitos dos fármacosRESUMO
In vitro culturing and mechanical properties of three types of three-dimensional poly(caprolactone) scaffolds with interconnecting open-foam networks are reported. The scaffolds targeted bone tissue regeneration and were fabricated using twin screw extrusion and coextrusion techniques, for continuous mixing/shaping and formation of single or multilayers with distinct and tailorable porosities and pore sizes. Human fetal preosteoblastic cells, hFOB, were cultured on the extruded and coextruded scaffolds under osteogenic supplements and the samples of the resulting tissue constructs were removed and characterized for cell viability and proliferation using the MTS assay, differentiation, and mineralized matrix synthesis via the alkaline phosphatase, ALP, activity and Alizarin Red staining and cell migration using confocal microscopy and scanning electron microscopy. The hFOB cells formed a confluent lining on scaffold surfaces, migrated to the interior and generated abundant extracellular matrix after 2 weeks of culturing, indicative of the promise of such scaffolds for utilization in tissue engineering. The scaffolds and tissue constructs exhibited compressive fatigue behavior that was similar to that of cancellous bone, suggesting the suitability of their use as bone graft substitutes especially for repair of critical-sized defects or nonunion fractures.
Assuntos
Parafusos Ósseos , Osso e Ossos/fisiologia , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Osteoblastos/citologia , Poliésteres/farmacologia , Engenharia Tecidual/métodos , Fosfatase Alcalina/metabolismo , Fenômenos Biomecânicos/efeitos dos fármacos , Osso e Ossos/efeitos dos fármacos , Cálcio/metabolismo , Comunicação Celular/efeitos dos fármacos , Contagem de Células , Células Cultivadas , Força Compressiva/efeitos dos fármacos , Feto/citologia , Humanos , Microscopia Confocal , Microscopia Eletrônica de Varredura , Osteoblastos/efeitos dos fármacos , Osteoblastos/enzimologia , Estresse Mecânico , Alicerces Teciduais/químicaRESUMO
We have created hyaluronic acid (HA)-based, cell-adhesive hydrogels that direct the initial attachment and the subsequent differentiation of human mesenchymal stem cells (MSCs) into pre-osteoblasts without osteogenic supplements. HA-based hydrogel particles (HGPs) with an average diameter of 5-6 µm containing an estimated 2.2 wt% gelatin (gHGPs) were synthesized by covalent immobilization of gelatin to HA HGPs prepared via an inverse emulsion polymerization technique. Separately, a photocrosslinkable HA macromer (HAGMA) was synthesized by chemical modification of HA with glycidyl methacrylate (GMA). Doubly crosslinked networks (DXNs) were engineered by embedding gHGPs in a secondary network established by HAGMA at a particle concentration of 2.5 wt%. The resultant composite gels, designated as HA-gHGP, have an average compressive modulus of 21 kPa, and are non-toxic to the cultured MSCs. MSCs readily attached to these gels, exhibiting an early stage of stress fiber assembly 3 h post seeding. By day 7, stellate-shaped cells with extended filopodia were found on HA-gHGP gels. Moreover, cells had migrated deep into the matrix, forming a three dimensional, branched and interconnected cell community. Conversely, MSCs on the control gels lacking gelatin moieties formed isolated spheroids with rounded cell morphology. After 28 days of culture on HA-gHGP, Type I collagen production and mineral deposition were detected in the absence of osteogenic supplements, suggesting induction of osteogenic differentiation. In contrast, cells on the control gels expressed markers for adipogenesis. Overall, the HA-gHGP composite matrix has great promise for directing the osteogenic differentiation of MSCs by providing an adaptable environment through the spatial presentation of cell-adhesive modules.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Reagentes de Ligações Cruzadas/farmacologia , Gelatina/farmacologia , Ácido Hialurônico/farmacologia , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Animais , Anticorpos/farmacologia , Bovinos , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Fluorescamina/metabolismo , Humanos , Receptores de Hialuronatos/metabolismo , Integrinas/imunologia , Fenômenos Mecânicos/efeitos dos fármacos , Microscopia de Contraste de Fase , Tamanho da Partícula , FenótipoRESUMO
Polypropylene sutures (PP) are already used in surgery. Because microbial infection leads to complications, we developed antimicrobial PP suture by plasma-induced graft polymerization of acrylic acid followed by chitosan binding on the remaining carboxyl groups. Mechanical properties and surface morphologies were analyzed on these sutures. Tetracycline hydrochloride (TC) or nanosilver (NS) was then immobilized to PP. The resulting PP sutures evidenced drug release properties and antimicrobial activity in vitro. PP implanted in vivo for 30 days in the muscle of rats showed the absence of adverse effects and a tissue organization. This new polypropylene suture with suitable antimicrobial features appears to be a promising macromolecular material for clinical and cosmetic applications.
Assuntos
Materiais Biocompatíveis/síntese química , Teste de Materiais/métodos , Gases em Plasma/química , Polipropilenos/síntese química , Suturas , Células 3T3 , Animais , Anti-Infecciosos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Quitosana/química , Escherichia coli/efeitos dos fármacos , Masculino , Fenômenos Mecânicos/efeitos dos fármacos , Camundongos , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Polipropilenos/química , Implantação de Prótese , Ratos , Ratos Wistar , Staphylococcus aureus/efeitos dos fármacos , Propriedades de Superfície/efeitos dos fármacos , Tetraciclina/farmacologiaRESUMO
The marine diazotrophic cyanobacterium Trichodesmium responds to elevated atmospheric CO(2) partial pressure (pCO(2)) with higher N(2) fixation and growth rates. To unveil the underlying mechanisms, we examined the combined influence of pCO(2) (150 and 900 microatm) and light (50 and 200 micromol photons m(-2) s(-1)) on Trichodesmium IMS101. We expand on a complementary study that demonstrated that while elevated pCO(2) enhanced N(2) fixation and growth, oxygen evolution and carbon fixation increased mainly as a response to high light. Here, we investigated changes in the photosynthetic fluorescence parameters of photosystem II, in ratios of the photosynthetic units (photosystem I:photosystem II), and in the pool sizes of key proteins involved in the fixation of carbon and nitrogen as well as their subsequent assimilation. We show that the combined elevation in pCO(2) and light controlled the operation of the CO(2)-concentrating mechanism and enhanced protein activity without increasing their pool size. Moreover, elevated pCO(2) and high light decreased the amounts of several key proteins (NifH, PsbA, and PsaC), while amounts of AtpB and RbcL did not significantly change. Reduced investment in protein biosynthesis, without notably changing photosynthetic fluxes, could free up energy that can be reallocated to increase N(2) fixation and growth at elevated pCO(2) and light. We suggest that changes in the redox state of the photosynthetic electron transport chain and posttranslational regulation of key proteins mediate the high flexibility in resources and energy allocation in Trichodesmium. This strategy should enable Trichodesmium to flourish in future surface oceans characterized by elevated pCO(2), higher temperatures, and high light.
Assuntos
Dióxido de Carbono/farmacologia , Cianobactérias/efeitos dos fármacos , Cianobactérias/efeitos da radiação , Luz , Fenômenos Mecânicos , Fixação de Nitrogênio/efeitos dos fármacos , Fixação de Nitrogênio/efeitos da radiação , Aclimatação/efeitos dos fármacos , Aclimatação/efeitos da radiação , Proteínas de Bactérias/metabolismo , Dióxido de Carbono/metabolismo , Clorofila/metabolismo , Cianobactérias/enzimologia , Transporte de Elétrons/efeitos dos fármacos , Transporte de Elétrons/efeitos da radiação , Metabolismo Energético/efeitos dos fármacos , Fluorescência , Fenômenos Mecânicos/efeitos dos fármacos , Fenômenos Mecânicos/efeitos da radiação , Nitrogenase/metabolismo , Fotossíntese/efeitos dos fármacos , Fotossíntese/efeitos da radiação , Complexo de Proteína do Fotossistema II , Subunidades Proteicas/metabolismoRESUMO
Chitosan and collagen type I are naturally derived materials used as cell carriers because of their ability to mimic the extracellular environment and direct cell function. In this study beta-glycerophosphate (beta-GP), an osteogenic medium supplement and a weak base, was used to simultaneously initiate gelation of pure chitosan, pure collagen, and chitosan-collagen composite materials at physiological pH and temperature. Adult human bone marrow-derived stem cells (hBMSC) encapsulated in such hydrogels at chitosan/collagen ratios of 100/0, 65/35, 25/75, and 0/100 wt% exhibited high viability at day 1 after encapsulation, but DNA content dropped by about half over 12 days in pure chitosan materials while it increased twofold in materials containing collagen. Collagen-containing materials compacted more strongly and were significantly stiffer than pure chitosan gels. In monolayer culture, exposure of hBMSC to beta-GP resulted in decreased cell metabolic activity that varied with concentration and exposure time, but washing effectively removed excess beta-GP from hydrogels. The presence of chitosan in materials resulted in higher expression of osterix and bone sialoprotein genes in medium with and without osteogenic supplements. Chitosan also increased alkaline phosphatase activity and calcium deposition in osteogenic medium. Chitosan-collagen composite materials have potential as matrices for cell encapsulation and delivery, or as in situ gel-forming materials for tissue repair.
Assuntos
Osso e Ossos/fisiologia , Quitosana/farmacologia , Colágeno/farmacologia , Glicerofosfatos/farmacologia , Hidrogéis/farmacologia , Temperatura , Engenharia Tecidual/métodos , Fosfatase Alcalina/metabolismo , Animais , Osso e Ossos/efeitos dos fármacos , Bovinos , Morte Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , DNA/metabolismo , Géis , Regulação da Expressão Gênica/efeitos dos fármacos , Glicerofosfatos/toxicidade , Humanos , Teste de Materiais , Fenômenos Mecânicos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Soluções , Espectrometria por Raios X , Células-Tronco/citologia , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismoRESUMO
Healing large bone defects and non-unions remains a significant clinical problem. Current treatments, consisting of auto and allografts, are limited by donor supply and morbidity, insufficient bioactivity and risk of infection. Biotherapeutics, including cells, genes and proteins, represent promising alternative therapies, but these strategies are limited by technical roadblocks to biotherapeutic delivery, cell sourcing, high cost, and regulatory hurdles. In the present study, the collagen-mimetic peptide, GFOGER, was used to coat synthetic PCL scaffolds to promote bone formation in critically-sized segmental defects in rats. GFOGER is a synthetic triple helical peptide that binds to the alpha(2)beta(1) integrin receptor involved in osteogenesis. GFOGER coatings passively adsorbed onto polymeric scaffolds, in the absence of exogenous cells or growth factors, significantly accelerated and increased bone formation in non-healing femoral defects compared to uncoated scaffolds and empty defects. Despite differences in bone volume, no differences in torsional strength were detected after 12 weeks, indicating that bone mass but not bone quality was improved in this model. This work demonstrates a simple, cell/growth factor-free strategy to promote bone formation in challenging, non-healing bone defects. This biomaterial coating strategy represents a cost-effective and facile approach, translatable into a robust clinical therapy for musculoskeletal applications.