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1.
Chemistry ; 25(61): 13994-14002, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31506999

RESUMO

Quinone methide (QM) as a latent trapping unit has been widely explored in activity-based self-immobilizing reagents. However, further application of this strategy has been largely hampered by the limited labeling efficiency to proteins. In this study, a thorough investigation on the labeling efficiency and the structure of QM-based trapping unit is presented, from which a QM with multiple leaving groups was identified as an optimal trapping unit. An alkaline phosphatase (ALP) immobilizing reagent featured with this multiple-labeling trapping unit exhibited lower nonspecific binding and, remarkably, a significantly higher labeling efficiency over other immobilizing reagents upon enzymatic activation. The utility of this imaging reagent was further demonstrated with the in vitro and in vivo visualization of the ALP activities. Furthermore, the multiple functional trapping unit may find greater value in the other activity-based immobilizing probes.


Assuntos
Fosfatase Alcalina/metabolismo , Indolquinonas/química , Fosfatase Alcalina/química , Animais , Corantes Fluorescentes/química , Células HEK293 , Células HeLa , Humanos , Indolquinonas/metabolismo , Camundongos , Camundongos Nus , Microscopia de Fluorescência , Imagem Óptica , Coloração e Rotulagem , Transplante Heterólogo
2.
Int J Nanomedicine ; 14: 6151-6163, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31447557

RESUMO

Background: Precise control and induction of the differentiation of stem cells to osteoblasts by artificial biomaterials are a promising strategy for rapid bone regeneration and reconstruction. Purpose: In this study, gold nanoparticles (AuNPs)-loaded hydroxyapatite (HA-Au) nanocomposites were designed to guide the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs) through the synergistic effects of both AuNPs and HA. Materials and methods: The HA-Au nanoparticles were synthesized and characterized by several analytical techniques. Cell viability and proliferation of hMSCs were characterized by CCK-8 test. Cellular uptake of nanoparticles was observed by transmission electron microscope. For the evaluation of osteogenic differentiation, alkaline phosphatase (ALP) activity and staining, Alizarin red staining, and a quantitative real-time polymerase chain reaction (RT-PCR) analysis were performed. In order to examine specific signaling pathways, RT-PCR and Western blotting assay were performed. Results: The results confirmed the successful synthesis of HA-Au nanocomposites. The HA-Au nanoparticles showed good cytocompatibility and internalized into hMSCs at the studied concentrations. The increased level of ALP production, deposition of calcium mineralization, as well as the expression of typical osteogenic genes, indicated the enhancement of osteogenic differentiation of hMSCs. Moreover, the incorporation of Au could activate the Wnt/ß-catenin signaling pathway, which seemed to be the molecular mechanism underlying the osteoinductive capability of HA-Au nanoparticles. Conclusion: The HA-Au nanoparticles exerted a synergistic effect on accelerating osteogenic differentiation of hMSCs, suggesting they may be potential candidates for bone repair and regeneration.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Durapatita/farmacologia , Ouro/química , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Nanopartículas Metálicas/química , Osteogênese , Via de Sinalização Wnt/efeitos dos fármacos , Fosfatase Alcalina/metabolismo , Cálcio/metabolismo , Diferenciação Celular/genética , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/enzimologia , Nanopartículas Metálicas/ultraestrutura , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteogênese/genética
3.
Int J Nanomedicine ; 14: 4975-4989, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31371942

RESUMO

The porous surface of a polyetheretherketone (PK)-nanoporous lithium-doped magnesium silicate (NLS) blend (PKNLS) was fabricated on a PK surface by layer-by-layer pressuring, sintering, and salt-leaching. As controls, porous surfaces of a PK/lithium-doped magnesium silicate blend (PKLS) and PK were fabricated using the same method. The results revealed that porosity, water absorption, and protein absorption of the porous surface of PKNLS containing macropores and nanopores were obviously enhanced compared to PKLS and PK containing macropores without nanopores. In addition, PKNLS, with both macroporostiy and nanoporosity, displayed the highest ability of apatite mineralization in simulated body liquid, indicating excellent bioactivity. In vitro responses (including adhesion, proliferation, and differentiation) of MC3T3E1 cells to PKNLS were significantly enhanced compared to PKLS and PK. In vivo implantation results showed that new bone grew into the macroporous surface of PKNLS, and the amount of new bone for PKNLS was the highest. In short, PKNLS integration with PK significantly promoted cells/bone-tissue responses and exhibited excellent osteogenesis in vivo, which might have great potential for bone repair.


Assuntos
Osso e Ossos/fisiologia , Cetonas/farmacologia , Lítio/farmacologia , Silicatos de Magnésio/farmacologia , Nanoporos , Osteoblastos/citologia , Polietilenoglicóis/farmacologia , Adsorção , Fosfatase Alcalina/metabolismo , Animais , Apatitas/química , Osso e Ossos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Imagem Tridimensional , Masculino , Camundongos , Nanoporos/ultraestrutura , Osteoblastos/efeitos dos fármacos , Osteoblastos/ultraestrutura , Osteogênese/efeitos dos fármacos , Porosidade , Coelhos , Água/química , Difração de Raios X
4.
Oncology ; 97(4): 236-244, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31412345

RESUMO

INTRODUCTION: On a global scale, the malignant growth of mammary gland is the most common type of cancer in women. In the progress of mammary carcinoma, osseous metastatic invasion has a pivotal significance because it is a frequent complication occurring at an early stage of the disease. BACKGROUND: Bone metastases in breast cancer patients lead to increased mortality and decreased health-related quality of life. Therefore, early diagnostic assessment and treatment is requested. Meanwhile the progress of the disease should be monitored closely. Regarding health-related quality of life and lifetime prolongation, osseous metastases should be early diagnosed, therapied, and monitored. Up to date the gold standard is the whole-body scintigraphy. This kind of bone imaging features has high sensitivity but shows loss of specificity. AIM: This study aims to investigate the diagnostic versatility of bone markers in its resorption and formation function to detect bone metastases in patients with breast cancer. PATIENTS, MATERIALS, AND METHODS: For this purpose, the concentration of competing bone processing tumor markers in serums of 78 patients was detected and analyzed. Two groups of women with mammary carcinoma with and without osseous metastases were built to examine the presence (or absence) of statistically significant disparity of tumor marker concentration. The tumor markers employed in this study were the carboxyterminal collagen type I telopeptid (CTX), known as beta-crosslaps (ß-CTx), the alkaline phosphatase (AP), and its isoenzymes (especially the bone-specific AP [B-AP]). Additionally, the tumor markers for breast cancer (CA 15-3 and CEA) were analyzed in both groups. RESULTS: Our results provide evidence that in both groups, tumor markers such as ß-CTx and B-AP were a promising tool for the detection and exclusion of bone metastases in breast cancer. This comprehensive investigation shows both ß-CTx and B-AP are able to fulfill the conditions of a competent appliance to detect osseous metastases of patients with mammary carcinoma. CONCLUSION: Concerning the urgency of early and frequent detection, staging, and disease monitoring of mammary carcinoma with osseous metastases, this study renewed and underlined the importance of biochemical tumor markers - especially ß-CTx and B-AP - and laid a clinical-based cornerstone to build up on a prospective research.


Assuntos
Biomarcadores/metabolismo , Neoplasias Ósseas/metabolismo , Osso e Ossos/metabolismo , Neoplasias da Mama/metabolismo , Fosfatase Alcalina/metabolismo , Biomarcadores Tumorais , Neoplasias Ósseas/secundário , Neoplasias da Mama/patologia , Colágeno Tipo I/metabolismo , Feminino , Humanos , Mucina-1/metabolismo , Metástase Neoplásica , Qualidade de Vida , Curva ROC , Cintilografia , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Imagem Corporal Total
5.
Pestic Biochem Physiol ; 158: 156-165, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31378352

RESUMO

Culex pipiens is a main vector for Bancroftian filariasis, Rift Valley Fever and diseases caused by other viruses, leaving several peoples with disabilities. In recent years, plant derived compounds have received much attention as potential alternatives to synthetic chemicals due to their low toxicity to mammals and environmental persistence. Twenty-one monoterpenes from different chemical groups (hydrocarbons and oxygenated products) were evaluated against Culex pipiens larvae. In addition, in vivo biochemical studies including effects on acetylcholine esterase (AChE), acid and alkaline phosphatases (ACP and ALP), total adenosine triphosphatase (ATPase) and gamma-aminobutyric acid transaminase (GABA-T) were investigated. Furthermore, in silico studies including pharmacophore elucidation, ADMET analysis and molecular docking of these compounds were performed. Among all tested monoterpenes, hydrocarbons [p-cymene, (R)-(+)-limonene and (+)-α-pinene], acetates (cinnamyl acetate, citronellyl acetate, eugenyl acetate and terpinyl acetate), alcohols [(±)-ß-citronellol and terpineol], aldehydes [citral and (1R)-(-)-myrtenal] and ketone [(R)-(+)-pulegone] exhibited the highest larval toxicity with LC50 = 14.88, 27.97, 26.13, 2.62, 3.81, 2.74, 21.65, 1.64, 21.70, 21.76, 1.68 and 1.90 mg/L after 48 h of exposure, respectively. The compounds proved a significant inhibition of all tested enzymes except total ATPase. The biochemical and molecular docking studies proved that AChE and GABA-T were the main targets for the tested monoterpenes.


Assuntos
Culex/virologia , Inseticidas/farmacologia , Monoterpenos/farmacologia , Fosfatase Alcalina/metabolismo , Animais , Culex/patogenicidade , Filariose Linfática/transmissão , Ativação Enzimática/efeitos dos fármacos , Esterases/metabolismo , Simulação de Acoplamento Molecular , Transaminases/metabolismo
7.
Bioelectromagnetics ; 40(6): 412-421, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31338867

RESUMO

Pulsed electromagnetic fields (PEMFs) have been shown to be a noninvasive physical stimulant for bone fracture healing. However, PEMF stimulation requires a relatively long period of time and its mechanism of action has not yet been fully clarified. Recently, the mammalian target of rapamycin (mTOR) pathway has been shown to be involved in bone formation. This study aimed to investigate the effects of PEMFs on osteoblastic MC3T3-E1 cells by examining various cellular responses including changes in the mTOR pathway. Continuous PEMF stimulation induced a transient phosphorylation of the mTOR pathway, whereas intermittent PEMF stimulation (1 cycle of 10 min stimulation followed by 20 min of stimulation pause) revitalized the reduced phosphorylation. Moreover, PEMF stimulation stimulated cell proliferation (bromodeoxyuridine incorporation) rather than differentiation (alkaline phosphatase activity), with a more notable effect in the intermittently stimulated cells. These results suggest that intermittent PEMF stimulation may be effective in promoting bone fracture healing by accelerating cell proliferation, and in shortening stimulation time. Bioelectromagnetics. 2019;40:412-421. © 2019 Bioelectromagnetics Society.


Assuntos
Campos Eletromagnéticos , Osteoblastos/citologia , Serina-Treonina Quinases TOR/metabolismo , Fosfatase Alcalina/metabolismo , Bromodesoxiuridina/metabolismo , Linhagem Celular , Proliferação de Células , Estimulação Elétrica , Humanos , Fosforilação , Transdução de Sinais
8.
Arch Insect Biochem Physiol ; 102(1): e21591, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31257641

RESUMO

In Brazil, the use of transgenic plants expressing the insect-toxic Bacillus thuringiensis endotoxin has been successfully used as pest control management since 2013 in transgenic soybean lineages against pest caterpillars such as Helicoverpa armigera. These toxins, endogenously expressed by the plants or sprayed over the crops, are ingested by the insect and bind to receptors in the midgut of these animals, resulting in disruption of digestion and lower insect survival rates. Here, we identified and characterized a membrane-associated alkaline phosphatase (ALP) in the midgut of Anticarsia gemmatalis, the main soybean defoliator pest in Brazil, and data suggested that it binds to Cry1Ac toxin in vitro. Our data showed a peak of ALP activity in homogenate samples of the midgut dissected from the 4th and 5th instars larvae. The brush border membrane vesicles obtained from the midgut of these larvae were used to purify a 60 kDa ALP, as detected by in-gel activity and in vitro biochemical characterization using pharmacological inhibitors and mass spectrometry. When Cry1Ac toxin was supplied to the diet, it was efficient in decreasing larval weight gain and survival. Indeed, in vitro incubation of Cry1Ac toxin with the purified ALP resulted in a 43% decrease in ALP specific activity and enzyme-linked immunosorbent assay showed that ALP interacts with Cry1Ac toxin in vitro, thus suggesting that ALP could function as a Cry toxin ligand. This is a first report characterizing an ALP in A. gemmatalis.


Assuntos
Fosfatase Alcalina/metabolismo , Proteínas de Bactérias/metabolismo , Endotoxinas/metabolismo , Proteínas Hemolisinas/metabolismo , Larva/enzimologia , Mariposas/enzimologia , Fosfatase Alcalina/antagonistas & inibidores , Fosfatase Alcalina/isolamento & purificação , Animais , Proteínas de Bactérias/toxicidade , Endotoxinas/toxicidade , Trato Gastrointestinal/enzimologia , Trato Gastrointestinal/ultraestrutura , Proteínas Hemolisinas/toxicidade , Microvilosidades/enzimologia
9.
Pan Afr Med J ; 32: 169, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31303938

RESUMO

A 61-year old female patient who was referred to the endocrine clinic for evaluation of an elevated alkaline phosphatase. She was originally referred to gastroenterology (GI), however no GI causes of elevated alkaline phosphatase was found. Upon fractionation, it was noted that she had elevation in bone specific alkaline phosphatase. Past history was significant for hypertension, atrial fibrillation and menopause 6 years ago. She was also noted to have multiple drug allergies manifesting as urticaria and flushing. Review of the past records revealed a persistently elevated alkaline phosphatase over the last two years. She had no history of falls or fractures. Computed tomography (CT) abdomen done to rule out biliary pathology, revealed osteosclerotic and osteolytic lesion in the pelvis concerning neoplastic disease. Bone marrow biopsy however, was negative for cancer but consistent with systemic mastocytosis (SM). Dual Energy X-ray absorbimetery (DEXA) scan revealed osteoporosis Serum tryptase levels were elevated; further genetic analysis showed a positive CKIT D816 mutation. She was started on bisphosphonates (initially alendronate and then ibandronate). Upon follow up at two years she had not experienced any fractures and her bone mineral density also had improved significantly.


Assuntos
Fosfatase Alcalina/metabolismo , Mastocitose Sistêmica/diagnóstico , Osteoporose/etiologia , Absorciometria de Fóton/métodos , Densidade Óssea , Difosfonatos/administração & dosagem , Feminino , Humanos , Mastocitose Sistêmica/complicações , Pessoa de Meia-Idade , Osteoporose/diagnóstico , Osteoporose/tratamento farmacológico
10.
Environ Pollut ; 252(Pt B): 1429-1438, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31265953

RESUMO

The aim of the work was to determine the trend, intensity and changes of selected microbial and phytotoxic parameters of degraded soil in the area of former sulphur mine reclaimed by post-flotation lime (PFL), sewage sludge (SS), mineral wool (MW- mixed with soil, MWP-pad) and mineral fertilizer (NPK). The following parameters: number of proteolytic bacteria and fungi, ammonification, nitrification, activities of alkaline phosphatase and arylsulphatase Lepidium sativum growth index (GI) and phenolic compounds were analysed in the soil in second and third year of the experiment. The addition of the SS separately or in combination with other remediation agents was found to be the most valuable for the number of microorganisms, intensification of nitrification process and enzymatic activities. In objects where other materials were added without sewage sludge, the inhibition of fungal growth as well as alkaline phosphatase and arylsulphatase activities was observed, however the inhibitory effect declined with time. The observed increase of GI shows the long-term, positive effect of treatments on soil properties concerning plant growth. The use of lime and lime together with sewage sludge contributed to the decrease in the content of phenolic compounds in the reclaimed soil.


Assuntos
Bactérias/crescimento & desenvolvimento , Biodegradação Ambiental , Fungos/crescimento & desenvolvimento , Lepidium sativum/crescimento & desenvolvimento , Esgotos/química , Poluentes do Solo/análise , Enxofre/análise , Fosfatase Alcalina/metabolismo , Arilsulfatases/metabolismo , Fertilizantes/análise , Nitrificação/fisiologia , Fenol/análise , Solo/química , Microbiologia do Solo
11.
Georgian Med News ; (290): 12-16, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-31322506

RESUMO

Aim - study of marker enzymes, hormonal and carbohydrate-protein indicators of the state of reparative osteogenesis in patients with complicated and uncomplicated course of injuries of facial cranium. The study included 81 patients with injuries of facial cranium, which were divided into 2 groups, depending on the presence of complications. The following enzyme indicators were studied: the level of excretion of hydroxyproline in daily urine; alkaline and acid phosphatase activity; the percentage of bone isoenzymes of alkaline phosphatase. To assess the mineral metabolism, the level of total and ionized calcium and inorganic phosphorus in the blood serum, as well as their excretion in the urine, were determined. To assess the state of metabolism, the concentration of glycosaminoglycans and their fractions in the blood serum were studied. To study the structural and functional state of the bone tissue the densitometry was performed. In patients with complicated course of injuries of facial cranium assosiated with traumatic brain injury there was revealed the increase (р<0,05) of: excretion of phosphorus, uronic acids and oxyproline, while the excretion of calcium was not disturbed (р>0,05), and excretion of magnesium was decreased (р<0,05). It was found out that the level of calcium of blood serum in patients with complicated course is significantly (р<0,05) lower than in the control group and does not depend on the presence of craniocerebral injury (р>0,05). The decrease of the level of ionized calcium content in blood serum can be the confirmation of lower metabolic activity of reparative osteogenesis processes, first of all at the expense of damage of central mechanisms. When studying the content of carbohydrate-protein metabolites by complicated course of injuries of facial cranium, the absolute increase (р<0,05) of concentration of chondroitin-6-sulfates was revealed, and during the analysis of results it was found out that in absolute values, as well as in structural indexes, the specific weight of various fractions changes, that can be the evidence of instability of mechanisms of osteogenesis and of damage of physiological mechanisms of reparative osteogenesis. Densitometric equivalents of forming of complicated course of injuries of facial cranium are the increase of broadband ultrasonic attenuation and the decrease of its spreading speed on the background of low levels of chondroitin-6-sulfates.


Assuntos
Cálcio/sangue , Traumatismos Craniocerebrais , Traumatismos Faciais , Osteogênese/fisiologia , Fósforo/sangue , Crânio/lesões , Fosfatase Ácida/metabolismo , Fosfatase Alcalina/metabolismo , Cálcio/urina , Traumatismos Craniocerebrais/epidemiologia , Traumatismos Craniocerebrais/metabolismo , Traumatismos Faciais/enzimologia , Traumatismos Faciais/metabolismo , Glicosaminoglicanos/metabolismo , Humanos , Hidroxiprolina/urina , Minerais/metabolismo , Fósforo/urina
12.
Nat Commun ; 10(1): 2704, 2019 06 20.
Artigo em Inglês | MEDLINE | ID: mdl-31221964

RESUMO

Attachment of lipid tails to oligonucleotides has emerged as a powerful technology in constructing cell membrane-anchorable nucleic acid-based probes. In practice, however, conventional lipid-conjugated oligonucleotides fail to distinguish among different cell membranes. Herein, a phosphorylated lipid-conjugated oligonucleotide (DNA-lipid-P) is reported for alkaline phosphatase (ALP)-dependent cell membrane adhesion. In the absence of ALP, DNA-lipid-P with its poor hydrophobicity shows only weak interaction with cell membrane. However, in the presence of the highly expressed plasma membrane-associated ALP, DNA-lipid-P is converted to lipid-conjugated oligonucleotide (DNA-lipid) by enzymatic dephosphorylation. As a result of such conversion, the generated DNA-lipid has greater hydrophobicity than DNA-lipid-P and is thus able to insert into cell membranes in situ. Accordingly, DNA-lipid-P enables selective anchoring on cell membranes with elevated ALP level. Since elevated ALP level is a critical index of some diseases and even cancers, DNA-lipid-P holds promise for cell membrane engineering and disease diagnostics at the molecular level.


Assuntos
Fosfatase Alcalina/metabolismo , Membrana Celular/metabolismo , Sondas Moleculares/metabolismo , Oligonucleotídeos/metabolismo , Linhagem Celular Tumoral , Humanos , Interações Hidrofóbicas e Hidrofílicas , Lipídeos/química , Microscopia Confocal/métodos , Microscopia de Fluorescência/métodos , Imagem Molecular/métodos , Sondas Moleculares/química , Oligonucleotídeos/química , Compostos Organofosforados/química , Fosforilação
13.
Int J Clin Pharmacol Ther ; 57(8): 402-407, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31232278

RESUMO

OBJECTIVE: To investigate the population pharmacokinetics of delayed methotrexate (MTX) excretion in children with acute lymphoblastic leukemia (ALL). MATERIALS AND METHODS: A total of 1,659 plasma concentration samples of MTX from 190 patients with 1 - 4 courses (plasma concentrations > 0.1 µmol/L) were collected in this study. The data analysis was performed using Phoenix NLME 1.3 software. The covariates included age, body surface area (BSA), body weight, alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine transaminase (ALT), total bilirubin (TBIL), and serum creatinine (SCr). The final model was validated by bootstrap resampling procedures (1,000 runs) and visual predictive check (VPC) method. RESULTS: The data were best described by a two-compartment linear pharmacokinetic model. The mean values of clearance (CL) and distribution volume (Vd) of MTX were 6.53 L/h and 67.88 L, respectively. Analysis of covariates showed that BSA influenced the CL of MTX. CONCLUSION: The final model was demonstrated as appropriate and effective for assessing the pharmacokinetic parameters of delayed MTX excretion in children with ALL.


Assuntos
Antimetabólitos Antineoplásicos/farmacocinética , Metotrexato/farmacocinética , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamento farmacológico , Alanina Transaminase/metabolismo , Fosfatase Alcalina/metabolismo , Aspartato Aminotransferases/metabolismo , Bilirrubina/sangue , Criança , Creatinina/sangue , Humanos
14.
Int J Nanomedicine ; 14: 3785-3797, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31239662

RESUMO

Background: Brushite (dicalcium phosphate dihydrate, DCPD) cement as a promising bioactive material for bone tissue engineering is widely used to treat defects. However, relatively poor mechanical properties of brushite cement limit its application in loadbearing conditions. The aim of this study is to investigate the effect of graphene oxide (GO) addition to the physical-mechanical-biological properties of brushite cement. Methods: The brushite types of cement were prepared by mixing ß-tricalcium phosphate [ß-TCP, Ca3 (PO4)2] and monocalcium phosphate monohydrate [MCPM, Ca(H2PO4)2. H2O]. GO was introduced at 0, 0.5, 2, and 5 wt.% with the liquid. MG63 cells were cultured on the GO/CPC surfaces to observe various cellular activities and hydroxyapatite (HA) mineralization. Results: Based on our results, GO/CPC composites exhibit improvement in compressive strength compared to pure CPC. New Ca-deficient apatite layer was deposited on the composite surface after immersing immersion in SBF for 7 and 14 days. Field emission scanning electron microscope (FESEM) images indicated that pure and GO incorporated brushite cement facilitates cell adhesion. CPC/GO was slightly toxic to cells such that high concentrations of GO decreased the cell viability. Besides, alkaline phosphatase (ALP) activity of cells was improved compared with the pure CPC. Conclusion: Our results highlight the role of graphene oxide that may have great potential in enabling the utility of graphene-based materials in various biomedical applications.


Assuntos
Cimentos para Ossos/química , Fosfatos de Cálcio/química , Grafite/química , Teste de Materiais , Fosfatase Alcalina/metabolismo , Adesão Celular , Linhagem Celular Tumoral , Sobrevivência Celular , Força Compressiva , Durapatita/química , Módulo de Elasticidade , Humanos , Concentração de Íons de Hidrogênio , Porosidade , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo , Difração de Raios X
15.
Int J Nanomedicine ; 14: 4133-4144, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31239672

RESUMO

Background: Although titanium dioxide nanotubes (TNTs) had great potential to promote osteogenesis, their weak bonding strength with titanium substrates greatly limited their clinical application. Purpose: The objective of this study was to maintain porosity and improve the stability of TNT coatings by preparing some micro-patterned mesoporous/nanotube (MP/TNT) structures via a photolithography-assisted anodization technology. Methods: The adhesion strength of different coatings was studied by ultrasonic cleaning machine and scratch tester. The early adhesion, spreading, proliferation and differentiation of MC3T3-E1 cells on different substrates were investigated in vitro by fluorescent staining, CCK8, alkaline phosphatase activity, mineralization and polymerase chain reaction assays, respectively. Results: Results of ultrasonic and scratch assays showed that the stability of TNTs (especially 125 nm) was significantly improved after being patterned with MP structures. In vitro cell assays further demonstrated that the insertion of MP structure into 125 nm TNT coating, which was denoted as MP125, could effectively improve the early adhesion, spreading and proliferation of surface MC3T3-E1 cells without damaging their osteogenic differentiation. Conclusion: We determined that the MP/TNT patterned samples (especially MP125) have excellent stability and osteogenesis properties, and may have better clinical application prospects.


Assuntos
Nanotubos/química , Osteogênese , Titânio/química , Adsorção , Fosfatase Alcalina/metabolismo , Animais , Adesão Celular/genética , Diferenciação Celular/genética , Linhagem Celular , Proliferação de Células/genética , Forma Celular/genética , Sobrevivência Celular/genética , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Fluorescência , Regulação da Expressão Gênica , Humanos , Camundongos , Minerais/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogênese/genética , Osteopontina/genética , Osteopontina/metabolismo , Porosidade , Água/química
16.
Int J Nanomedicine ; 14: 4229-4245, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31239677

RESUMO

Purpose: Gene therapies via Noggin small interfering (si)RNA (siNoggin) and bone morphogenetic protein (BMP)-2 plasmid DNA (pBMP-2) may be promising strategies for bone repair/regeneration, but their ideal delivery vectors, efficacy difference, and underlying mechanisms have not been explored, so these issues were probed here. Methods: This study used lipopolysaccharide-amine nanopolymersomes (LNPs), an efficient cytosolic delivery vector developed by the research team, to mediate siNoggin and pBMP-2 to transfect MC3T3-E1 cells, respectively. The cytotoxicity, cell uptake, and gene knockdown efficiency of siNoggin-loaded LNPs (LNPs/siNoggin) were studied, then the osteogenic-differentiation efficacy of MC3T3-E1 cells treated by LNPs/pBMP-2 and LNPs/siNoggin, respectively, were compared by measuring the expression of osteogenesis-related genes and proteins, alkaline phosphatase (ALP) activity, and mineralization of the extracellular matrix at all osteogenic stages. Finally, the possible signaling pathways of the two treatments were explored. Results: LNPs delivered siNoggin into cells efficiently to silence 50% of Noggin expression without obvious cytotoxicity. LNPs/siNoggin and LNPs/pBMP-2 enhanced the osteogenic differentiation of MC3T3 E1 cells, but LNPs/siNoggin was better than LNPs/pBMP-2. BMP/Mothers against decapentaplegic homolog (Smad) and glycogen synthase kinase (GSK)-3ß/ß-catenin signaling pathways appeared to be involved in osteogenic differentiation induced by LNPs/siNoggin, but GSK-3ß/ß-catenin was not stimulated upon LNPs/pBMP-2 treatment. Conclusion: LNPs are safe and efficient delivery vectors for DNA and RNA, which may find wide applications in gene therapy. siNoggin treatment may be a more efficient strategy to enhance osteogenic differentiation than pBMP-2 treatment. LNPs loaded with siNoggin and/or pBMP-2 may provide new opportunities for the repair and regeneration of bone.


Assuntos
Proteína Morfogenética Óssea 2/metabolismo , Proteínas de Transporte/metabolismo , Diferenciação Celular , Lipopolissacarídeos/farmacologia , Nanopartículas/química , Osteogênese , Polímeros/química , RNA Interferente Pequeno/administração & dosagem , Fosfatase Alcalina/metabolismo , Aminas/química , Animais , Diferenciação Celular/genética , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Matriz Extracelular/metabolismo , Glicogênio Sintase Quinase 3 beta/metabolismo , Camundongos , Minerais/química , Nanopartículas/toxicidade , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Plasmídeos/administração & dosagem , Transdução de Sinais/efeitos dos fármacos , Transfecção , beta Catenina/metabolismo
17.
Nat Commun ; 10(1): 2829, 2019 06 27.
Artigo em Inglês | MEDLINE | ID: mdl-31249296

RESUMO

Extracellular vesicles (EVs) are involved in the regulation of cell physiological activity and the reconstruction of extracellular environment. Matrix vesicles (MVs) are a type of EVs released by bone-related functional cells, and they participate in the regulation of cell mineralization. Here, we report bioinspired MVs embedded with black phosphorus (BP) and functionalized with cell-specific aptamer (denoted as Apt-bioinspired MVs) for stimulating biomineralization. The aptamer can direct bioinspired MVs to targeted cells, and the increasing concentration of inorganic phosphate originating from BP can facilitate cell biomineralization. The photothermal effect of the Apt-bioinspired MVs can also promote the biomineralization process by stimulating the upregulated expression of heat shock proteins and alkaline phosphatase. In addition, the Apt-bioinspired MVs display outstanding bone regeneration performance. Our strategy provides a method for designing bionic tools to study the mechanisms of biological processes and advance the development of medical engineering.


Assuntos
Vesículas Extracelulares/metabolismo , Fósforo/metabolismo , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Aptâmeros de Nucleotídeos/genética , Aptâmeros de Nucleotídeos/metabolismo , Biomineralização , Osso e Ossos/química , Osso e Ossos/citologia , Osso e Ossos/metabolismo , Vesículas Extracelulares/química , Feminino , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Osteoblastos/química , Osteoblastos/metabolismo , Fosfatos/metabolismo , Fósforo/química , Ratos
18.
Int J Nanomedicine ; 14: 3831-3843, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31213804

RESUMO

Purpose: On the basis of reasonable superposition of various surface treatment methods, alkali-treated titanium with nanonetwork structures (TNS) was coated with mussel adhesive protein (MAP) and named TNS-MAP. The aims were to optimize the biological properties of TNS, endue it with new properties, and enhance its utility in clinical dental applications. Methods: TNS disks were coated with MAP and the product surface was characterized. Its osteogenic properties were determined by evaluating its effects on cell adhesion, cell proliferation, the expression of osteogenesis-related genes, and in vivo experiments. Results: The treated materials showed excellent hydrophilicity, good surface roughness, and advantages of both TNS and MAP. TNS-MAP significantly promoted initial cell attachment especially after 15 mins and 30 mins. At every time point, cell adhesion and proliferation, the detection rate of osteogenesis-related markers in the extracellular matrix, and the expression of osteogenesis-related genes were markedly superior on TNS-MAP than the control. The in vivo experiments revealed that TNS-MAP promoted new bone growth around the implants and the bone-implant interface. Conclusion: We verified through in vitro and in vivo experiments that we successfully created an effective TNS-MAP composite implant with excellent biocompatibility and advantages of both its TNS and MAP parent materials. Therefore, the new biocomposite implant material TNS-MAP may potentially serve in practical dentistry and orthopedics.


Assuntos
Álcalis/química , Materiais Revestidos Biocompatíveis/farmacologia , Nanopartículas/química , Osseointegração/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Proteínas/farmacologia , Titânio/farmacologia , Fosfatase Alcalina/metabolismo , Animais , Interface Osso-Implante/diagnóstico por imagem , Interface Osso-Implante/patologia , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Ratos Sprague-Dawley , Espectroscopia de Infravermelho com Transformada de Fourier , Microtomografia por Raio-X
19.
Int J Nanomedicine ; 14: 3929-3941, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31213809

RESUMO

Introduction: Hierarchical nanofibrous scaffolds are emerging as a promising bone repair material due to their high cell adhesion activity and nutrient permeability. However, the existing method for hierarchical nanofibrous scaffolds fabrication is complicated and not perfectly suitable for further biomedical application in view of both structure and function. In this study, we constructed a hierarchical nanofibrous poly (l-lactic acid)/poly(ε-caprolactone) (PLLA/PCL) scaffold and further evaluated its bone healing ability. Methods: The hierarchical PLLA/PCL nanofibrous scaffold (PLLA/PCL) was prepared by one-pot TIPS and then rapidly mineralized at room temperature by an electrochemical deposition technique. After electrode-positioning at 2 V for 2 hrs, a scaffold coated with hydroxyapatite (M-PLLA/PCL) could be obtained. Results: The pore size of the M-PLLA/PCL scaffold was hierarchically distributed so as to match the biophysical structure for osteoblast growth. The M-PLLA/PCL scaffold showed better cell proliferation and osteogenesis activity compared to the PLLA/PCL scaffold. Further in vivo bone repair studies indicated that the M-PLLA/PCL scaffold could accelerate defect healing in 12 weeks. Conclusion: The results of this study implied that the as-prepared hydroxyapatite coated hierarchical PLLA/PCL nanofibrous scaffolds could be developed as a promising material for efficient bone tissue repair after carefully tuning the TIPS and electrodeposition parameters.


Assuntos
Regeneração Óssea/fisiologia , Galvanoplastia/métodos , Minerais/química , Nanofibras/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/farmacologia , Tecidos Suporte/química , Fosfatase Alcalina/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Eletricidade , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/ultraestrutura , Osteogênese/efeitos dos fármacos , Porosidade , Ratos Sprague-Dawley , Crânio/diagnóstico por imagem , Crânio/efeitos dos fármacos , Crânio/patologia , Fatores de Tempo , Engenharia Tecidual/métodos , Microtomografia por Raio-X
20.
Food Chem Toxicol ; 131: 110540, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31173816

RESUMO

The effect of menaquinone-7 isolated from cheonggukjang was comparatively investigated with vitamin K1 and menaquinone-4 on cell differentiation and mineralization of the osteoblastic cell line MC3T3-E1. Results indicated that all vitamin K species significantly increased MC3T3-E1 cell proliferation, cellular alkaline phosphatase activity, osteocalcin synthesis, and calcium deposition in a dose-dependent manner. Menaquinone-4 and menaquinone-7 had more potent effects on calcium deposition than vitamin K1, and their effects were only partly reduced by warfarin (γ-carboxylation inhibitor) treatment, while warfarin abolished the induction activity of vitamin K1 on calcification. This suggests that vitamin K1 and K2 (menaquinone-4 & menaquinone-7) may have different mechanisms in stimulating osteoblast mineralization. In addition, the mRNA expression ratio of osteoprotegerin and the receptor activator of nuclear factor-kB ligand was also dramatically increased by treatment with vitamin K1 (62%), menaquinone-4 (247%), and menaquinone-7 (329%), suggesting that vitamin K may suppress the formation of osteoclast by up-regulating the ratio of osteoprotegerin/receptor activator of nuclear factor-kB ligand in osteoblasts. These results provide compelling evidence that vitamin K1, menaquinone-4, and menaquinone-7 all can promote bone health, which might be associated with elevations in the osteoprotegerin/receptor activator of nuclear factor-kB ligand ratio.


Assuntos
Biomineralização/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Alimentos de Soja , Vitamina K 1/farmacologia , Vitamina K 2/análogos & derivados , Fosfatase Alcalina/metabolismo , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Expressão Gênica/efeitos dos fármacos , Camundongos , Osteoblastos , Osteocalcina/metabolismo , Osteoprotegerina/genética , Ligante RANK/genética , Vitamina K 2/isolamento & purificação , Vitamina K 2/farmacologia
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