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1.
J Photochem Photobiol B ; 197: 111515, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31255939

RESUMO

An extraordinary arrangement of research is as yet going on in the area of orthopedic implants advancement to determine different issues being looked by the engineering today. In spite of a few detriments of the orthopedic metallic inserts, they keep on being utilized, essentially as a result of their unrivaled mechanical properties. We investigated the conceivable utilization of silicon carbide (SiC) as a nano-ceramic covering material of titanium (Ti)-based all out femoral substitution implants. The thought is to keep wear garbage arrangement from the delicate titanium exterior. Silicon carbide is a hard and firmly holding bio-ceramic surface substance, and in light of these physico-chemical properties, it isn't actually degradable, just like the case with apatite (HA). To improve cytocompatibility and osseous-integration, we deposited anodized titanium nanotubes (TiO2) inserts, by electrochemical deposition method (EDM), with silicon carbide (SiC) with apatite (SiC@HA). The deposition was affirmed by SEM, while phase composition properties were assessed by XRD. Calcium affidavit, osteocalcin creation, and articulation of bone genes were essentially higher in rodent osteoblast cell culture on SiC@HA-covered anodized titanium nanotubes than in cells cultured on uncoated anodized titanium nanotubes. Implantation into rodent femurs likewise demonstrated that the SiC@HA-covered substance had unrivaled osseous-integration movement in correlation with that of customary inserts, as evaluated by in vivo tomography and histology. Therefore, anodized titanium nanotubes covered with SiC@HA holds guarantee as an orthopedic implant substance.


Assuntos
Regeneração Óssea , Compostos Inorgânicos de Carbono/química , Materiais Revestidos Biocompatíveis/química , Durapatita/química , Nanopartículas/química , Compostos de Silício/química , Titânio/química , Animais , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/metabolismo , Osso e Ossos/patologia , Adesão Celular/efeitos dos fármacos , Materiais Revestidos Biocompatíveis/farmacologia , Materiais Revestidos Biocompatíveis/uso terapêutico , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Fraturas do Fêmur/terapia , Regulação da Expressão Gênica/efeitos dos fármacos , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteocalcina/metabolismo , Próteses e Implantes , Ratos
2.
J Biol Regul Homeost Agents ; 33(4): 1105-1111, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31332987

RESUMO

The adapter protein myeloid differentiation primary response gene 88 (MyD88) links the intracellular domains of interleukin receptors 1 and 18, and most Toll-like receptors (TLRs) to interleukin 1 receptor associated kinase (IRAK) signaling and subsequent NF-κB-mediated transcription. Previous work showed that mice with global deficiency of MyD88 (MyD88-/-) have osteopenic cancellous bone along with a reduction in osteoblastic but also osteoclastic surfaces. To further elucidate the role of MyD88 in bone, we utilized mice with osteoclast-restricted MyD88 expression in bone (MyD88OC). Bones of MyD88OC and wild type (wt) mice were examined by microCT analysis. Mechanical properties of bones were tested by three-point bending, and gene expression measured using quantitative real-time polymerase chain reaction. In MyD88OC mice, no osteopenic traits were observed, however, a drastic reduction in geometric parameters was detected. In trabecular bone a loss of connectivity density (-44%, p less than 0.0001) was measured and in cortical bone Imax (-31%, p less than 0.0001), Imin (-20%, p less than 0.001), J (-26%, p less than 0.0001) were reduced. Mechanical testing showed increased load to failure (77%, p less than 0.01) and decreased deflection at failure (-68%, p less than 0.01) of the femur. On the molecular level, relative gene expression analysis showed a (-29%, p less than 0.01) reduction in receptor activator of nuclear factor κ B ligand (RANKL) and no difference in osteoprotegerin (OPG) or RANK. Further, the bone resorption markers cathepsin K (CTSK) and tartrate-resistant acid phosphatase 5 (TRAP) were unchanged. In contrast, the bone formation markers collagen type 1 (COL1A1) and osteocalcin (OC) were decreased by -72% (p less than 0.0001) and -82% (p less than 0.0001), respectively. Together, our data suggests that the function of MyD88 in osteoclasts is sufficient to maintain bone mass, while it fails to preserve bone geometry, likely through dysfunctions in osteoblasts.


Assuntos
Reabsorção Óssea , Osso e Ossos/patologia , Fator 88 de Diferenciação Mieloide/metabolismo , Osteoclastos/citologia , Animais , Catepsina K/metabolismo , Diferenciação Celular , Colágeno Tipo I/metabolismo , Camundongos , Osteoblastos , Osteocalcina/metabolismo , Osteoclastos/metabolismo , Osteoprotegerina/metabolismo , Ligante RANK/metabolismo , Receptor Ativador de Fator Nuclear kappa-B/metabolismo , Fosfatase Ácida Resistente a Tartarato/metabolismo
3.
Int J Nanomedicine ; 14: 4881-4893, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31308664

RESUMO

Purpose: This study was designed to evaluate the in vitro and in vivo biocompatibility and osteointegration of plasma-sprayed hydroxyapatite (HA)-coated polyethylene terephthalate (PET) ligaments encapsulated with a simvastatin (SV)-chitosan (CS) composite. Methods: This study compared the in vitro and in vivo bone responses to three different PET ligaments: SV/CS/PET-HA, CS/PET-HA and PET-HA. A field emission scanning electron microscope was used to characterize the morphology, and the in vitro SV release profile was analyzed. MC3T3 cells were cocultured with SV/CS/PET-HA, CS/PET-HA and PET-HA to test their biocompatibility using CCK-8 tests. Osteogenic differentiation was investigated by the expression of marker genes using qPCR. Osteointegration was performed by implanting the PET ligaments into the proximal tibia bone tunnels of male Sprague-Dawley rats for 3 weeks and 6 weeks. The bone-implant interface was evaluated by micro-computed tomography (micro-CT) and histological analysis. Results: The characteristic nanoporous structures mainly formed on the surface of the plasma-sprayed HA particles in the SV/CS/PET-HA and CS/PET-HA groups. The SV release test showed that the sustained release of simvastatin lasted for 25 days in the SV/CS/PET-HA group. The in vitro studies demonstrated that the SV/CS/PET-HA ligaments induced osteogenic differentiation in the MC3T3 cells, with higher mRNA expression levels of collagen-1, bone morphogenetic protein-2, osteocalcin and alkaline phosphatase than those in the CS/PET-HA and PET-HA ligament groups. The in vivo tests showed that both micro-CT analysis (bone mineral density and bone volume per total volume) and histological analysis (bone implant contact and interface area) revealed significantly higher peri-implant bone formation and less interface area in the SV/CS/PET-HA group than in the other groups. Conclusion: The SV-CS composite nanoporous structure was associated with the improved biocompatibility and osteogenic differentiation in vitro and enhanced osteointegration process in vivo of plasma-sprayed HA-coated PET ligaments.


Assuntos
Quitosana/química , Durapatita/farmacologia , Ligamentos/efeitos dos fármacos , Nanoporos , Osseointegração/efeitos dos fármacos , Polietilenotereftalatos/farmacologia , Sinvastatina/farmacologia , Animais , Osso e Ossos/diagnóstico por imagem , Osso e Ossos/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Liberação Controlada de Fármacos , Masculino , Nanoporos/ultraestrutura , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogênese/efeitos dos fármacos , Osteogênese/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos Sprague-Dawley , Microtomografia por Raio-X
4.
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
5.
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
6.
Life Sci ; 232: 116582, 2019 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-31220525

RESUMO

AIMS: Vascular calcification/aging can cause different kind of serious diabetic vascular complications. High glucose could induce vascular smooth muscle cells (VSMCs) calcification/aging and then lead to diabetes-related vascular calcification/aging. In this study, we investigated how information in the blood is transmitted to VSMCs and the mechanisms of VSMCs calcification/aging under hyperglycaemic conditions. MATERIALS AND METHODS: Transmission electron microscopy and molecular size analysis were used to assess the morphology and size of exosomes. Alizarin Red S staining and senescence-associated ß galactosidase (SA-ß-gal) staining were carried out to detect calcification and senescence in VSMCs, respectively. Proteomics analysis was carried out to detect the different expression of exosomal proteins. Protein levels were measured by western blot analysis. KEY FINDINGS: The results show that exosomes isolated from high glucose stimulated human umbilical vein endothelial cell (HG-HUVEC-Exo) exhibited a bilayer structure morphology with a mean diameter of 63.63 ±â€¯2.96 nm. The presence of exosome markers including CD9, CD63 and TSG101 were also detected in HG-HUVEC-Exo. High glucose could induce VSMCs calcification/aging by increasing the expression of osteocalcin (OC) and p21 as well as the formation of mineralised nodules and SA-ß-gal positive cells. Fluorescence microscopy verified that the exosomes were taken up by VSMCs and Notch3 protein was enriched in HG-HUVEC-Exo. Most importantly, mTOR signalling was closely related to Notch3 protein and was involved in regulating HG-HUVEC-Exo-induced VSMCs calcification/aging. SIGNIFICANCE: The data demonstrate that Notch3 is required for HG-HUVEC-Exo promoted VSMCs calcification/aging and regulates VSMCs calcification/aging through the mTOR signalling pathway.


Assuntos
Músculo Liso Vascular/metabolismo , Receptor Notch3/fisiologia , Calcificação Vascular/metabolismo , Calcificação Fisiológica/efeitos dos fármacos , Cálcio/metabolismo , Células Cultivadas , Senescência Celular/fisiologia , Complicações do Diabetes/metabolismo , Complicações do Diabetes/fisiopatologia , Células Endoteliais/metabolismo , Células Endoteliais/fisiologia , Exossomos/metabolismo , Glucose/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Hiperglicemia/metabolismo , Músculo Liso Vascular/fisiologia , Miócitos de Músculo Liso/metabolismo , Osteocalcina/metabolismo , Receptor Notch3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Calcificação Vascular/fisiopatologia
7.
Cell Biol Int ; 43(5): 565-573, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30958604

RESUMO

Heterotopic ossification (HO) is a common disturbing complication of intra-articular fractures. Its prevention and treatment are still difficult as its pathogenesis is unclear. It was reported that PDGFRα+ muscle cells in skeletal muscle may participate in the formation of HO; however, the specific mechanism is still unknown. This study investigated the function of miR-19b-3p in osteogenic differentiation of PDGFRα+ muscle cells. MiR-19b-3p was upregulated during PDGFRα+ muscle cell osteogenic differentiation. The exogenous expression of miR-19b-3p led to an increase in osteogenic marker gene transcription and translation during the osteogenic differentiation of PDGFRα+ muscle cells. Furthermore, both alkaline phosphatase and alizarin red staining increased in miR-19b-3p mimic transfected cells. Over-expression of miR-19b-3p led to the down-regulation of gene of phosphate and tension homology deleted on chromosome ten (PTEN). Additionally, the dual luciferase reporter assay demonstrated that PTEN was a direct target of miR-19b-3p. The increase of osteocalcin, osteopontin, and Runt-related transcription factor 2 protein levels induced by ectopic miR-19b-3p expression could be partially reversed by PTEN over-expression. In conclusion, our results suggested that miR-19b-3p may be a promising target in inhibiting PDGFRα+ muscle cell osteogenic differentiation and treatment of HO.


Assuntos
MicroRNAs/metabolismo , Ossificação Heterotópica/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/metabolismo , Diferenciação Celular/fisiologia , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Regulação para Baixo , Humanos , MicroRNAs/genética , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Ossificação Heterotópica/genética , Ossificação Heterotópica/patologia , Osteocalcina/metabolismo , PTEN Fosfo-Hidrolase/genética , Receptor alfa de Fator de Crescimento Derivado de Plaquetas/genética
8.
Zhongguo Zhong Yao Za Zhi ; 44(3): 535-540, 2019 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-30989919

RESUMO

To investigate the preventive effect and possible mechanism of puerarin(Pur) in rat model of disuse osteoporosis(DOP),thirty healthy Wistar female rats of 2 months old were randomly divided into control group(Control), hindlimb suspension group(HLS), and puerarin group(HLS+Pur) in hindlimb suspension, with 10 rats in each group. A disuse osteoporosis model was established by tail suspension method, and 15.4 mg·kg~(-1) puerarin suspension was administered to HLS+Pur group every day, and the same volume of distilled water was administered to Control group and HLS group respectively. After 28 days, the rats were sacrificed by abdominal aorta blood collection, the main organs of the rats were removed, and the bone tissues of the rats were dissected. The organ index of the rats was calculated and the histopathology of the organs was observed under microscope. Bone mineral density test and bone biomechanical experiment were performed. Bone histomorphometry results were observed after bone tissue sectioning, and serum biochemical markers of bone metabolism were determined. There was no significant difference in organ index between the groups. There was no obvious abnormality in the pathological examination of the organs. The results of bone mineral density showed that puerarin could significantly increase the bone density of the tibia and vertebrae caused by hindlimb suspension. The mechanical parameters experiments showed that puerarin could effectively increase the maximum load and elastic modulus of the tibia and vertebrae. Fluorescence labeling showed that the fluorosis interval increased and the bone formation increased during puerarin treatment. The VG staining results showed that compared with the HLS group, in the puerarin group, the number of trabecular bone increased, the thickness of the trabecular bone became thicker, and the bone separation became smaller, which greatly improved the bone microstructure after hindlinb suspension. In addition, serum biochemical indicators showed that puerarin could promote bone formation index bone calcium. The content of osteocalcin(OC) increased and inhibited the formation of tartrate-resistant acid phosphatase 5 b(TRACP 5 b). Puerarin has a preventive effect in the rat model of disuse osteoporosis and its effect is good, and its mechanism may be related to promoting bone formation and inhibiting bone resorption.


Assuntos
Densidade Óssea , Isoflavonas/farmacologia , Osteoporose/tratamento farmacológico , Animais , Feminino , Osteocalcina/metabolismo , Ratos , Ratos Wistar , Fosfatase Ácida Resistente a Tartarato/metabolismo
9.
Nutrients ; 11(4)2019 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-30934881

RESUMO

Vitamin D is well known for its effects on calcium and mineral metabolism. However, vitamin D effects on bone turnover markers (BTMs), which are used together with bone mineral density (BMD) to evaluate bone health, are less clear. We therefore examined vitamin D effects on BTMs (beta-cross laps (CTX) and osteocalcin (OC)) and BMD in a post-hoc analysis of a randomized controlled trial (RCT). This is a post-hoc analysis of the Graz Vitamin D&TT-RCT, a single-center, double-blind, randomized placebo-controlled trial conducted between December 2012 and November 2017 at the endocrine outpatient clinic at the Medical University of Graz, Austria. A total of 200 healthy men with serum 25-hydroxyvitamin D (25(OH)D) levels <75 nmol/L participated in the trial. Subjects were randomized to receive 20,000 IU of vitamin D3/week (n = 100) or placebo (n = 100) for 12 weeks. Outcome measures were BTMs, BMD, and trabecular bone score (TBS). A total of 192 men (mean age and 25(OH)D: 43 (±13) years and 54.9 (±18.3) nmol/L, respectively) completed the study. We found no significant treatment effect on BTMs, BMD, or TBS (p > 0.05 for all). In middle-aged healthy men, vitamin D treatment for 12 weeks had no significant effect on BTMs or BMD.


Assuntos
Densidade Óssea/efeitos dos fármacos , Osso e Ossos/metabolismo , Suplementos Nutricionais , Vitamina D/administração & dosagem , Vitamina D/farmacologia , Adulto , Método Duplo-Cego , Humanos , Masculino , Pessoa de Meia-Idade , Osteocalcina/metabolismo , Estações do Ano , Testosterona/sangue , Testosterona/metabolismo
10.
Mater Sci Eng C Mater Biol Appl ; 100: 165-177, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30948050

RESUMO

The objective of this study was to examine behavior and function of osteoblasts on saliva-contaminated titanium and its potential improvement after UV light treatment. Acid-etched titanium disks were contaminated with human saliva. Osteoblasts derived from rat femur were cultured on contaminated and clean titanium disks. Contaminated disks further treated with UV light were also tested. The number of attached cells, the degree of cell spreading, and the expression of adhesion protein were significantly decreased on saliva-contaminated surfaces compared with clean surfaces. The gene expression of osteocalcin was also downregulated on contaminated surfaces, whereas ALP activity and mineralization were not significantly influenced. The impaired functions on contaminated surfaces were significantly increased if the surfaces were further treated with UV and even outperformed the ones on clean titanium surfaces. XPS analysis revealed that the atomic percentage of carbon and nitrogen detected on contaminated surfaces were substantially decreased after UV treatment. These results suggest that osteoblastic behavior and function were compromised on titanium surfaces contaminated with saliva. The compromised functions no longer happened if the surfaces were further treated with UV light, providing the basis to understand the effect of biological contamination on osseointegration and to explore UV treatment as a decontaminating technology.


Assuntos
Saliva/química , Titânio/química , Raios Ultravioleta , Animais , Apoptose/efeitos dos fármacos , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Adesão Celular/efeitos dos fármacos , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Humanos , Masculino , Microscopia Confocal , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Ratos , Ratos Sprague-Dawley , Propriedades de Superfície , Titânio/farmacologia
11.
Mater Sci Eng C Mater Biol Appl ; 100: 196-208, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30948053

RESUMO

Recently, natural polymers are reinforced with silica particles for hard tissue engineering applications to induce bone regeneration. In this study, as two novel bioactive agents, effects of diatomite and polyhedral oligomeric silsesquioxanes (POSS) on chitosan (CS)/Na-carboxymethylcellulose (Na-CMC) polymer blend scaffolds are examined. In addition, the effect of silica reinforcements was compared with Si-substituted nano-hydroxyapatite (Si-Hap) particles. The morphology, physical and chemical structures of the scaffolds were characterized with SEM, liquid displacement, FT-IR, mechanical analysis, swelling and degradation studies. The particle size and the crystal structure of diatomite, POSS and Si-Hap particles were determined with DLS and XRD analyses. In vitro studies were performed to figure out the cytotoxicity, proliferation, ALP activity, osteocalcin production and biomineralization to demonstrate the promising use of natural silica particles in bone regeneration. Freeze-dried scaffolds showed 190-307 µm pore size range and 61-70% porosity. Both inorganic reinforcements increased the mechanical strength, enhanced the water uptake capacity and fastened the degradation rate. The nanocomposite scaffolds did not show any cytotoxic effect and enhanced the surface mineralization in osteogenic medium. Thus, diatomite and POSS cage structures can be potential reinforcements for nanocomposite design in hard tissue engineering applications.


Assuntos
Regeneração Óssea , Carboximetilcelulose Sódica/química , Terra de Diatomáceas/química , Metacrilatos/química , Compostos de Organossilício/química , Polieletrólitos/química , Dióxido de Silício/química , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Regeneração Óssea/efeitos dos fármacos , Osso e Ossos/fisiologia , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Força Compressiva , Camundongos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteocalcina/metabolismo , Porosidade
12.
Pol J Vet Sci ; 22(1): 143-150, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30997775

RESUMO

Osteocalcin is a major non-collagenous component of the bone extracellular matrix and is considered to be an indicative factor of osteoblast differentiation. In the present study, we detected osteocalcin expression in different antler areas and growth phases by immunohisto- chemistry. Osteocalcin was highly expressed in all areas during the mineralization period and in mesenchymal cell and chondrocyte areas during the rapid growth period. The nucleotide sequence of the osteocalcin gene in sika deer antler was determined. The open reading frame was 303 bp encoding a protein of 100 amino acids. The estimated molecular mass of osteocalcin was 10.38 kDa and the theoretical isoelectric point was 5.37. The osteocalcin gene with a 6× His-tag at the C-terminus was cloned into the pGEX-4T1 vector and expressed in Escherichia coli under optimal conditions. The recombinant soluble protein fused with GST was purified with Ni-NTA resin. The purified osteocalcin protein exhibited a significant increase in HA adhesion and promoted antler chondrocyte proliferation. Osteocalcin is an important factor in regulating the rapid growth and differentiation of deer antlers.


Assuntos
Chifres de Veado/metabolismo , Clonagem Molecular , Cervos/fisiologia , Regulação da Expressão Gênica/fisiologia , Osteocalcina/metabolismo , Sequência de Aminoácidos , Animais , Masculino , Osteocalcina/genética
13.
Mater Sci Eng C Mater Biol Appl ; 99: 1174-1181, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30889651

RESUMO

Titanium (Ti) and its alloys are mainly used for dental and orthopedic applications due to their excellent biocompatibility and mechanical properties. However, their intrinsic bioinertness often quotes as a common complaint for biomedical applications. Herein, we produced nanopattern Ti surfaces with 10 nm nanopores in 120 nm dimples by electrochemical nanopattern formation (ENF), and evaluated the osteogenic differentiation of human mesenchymal stem cells (hMSCs) on the nanopattern Ti surfaces. The ENF surfaces were obtained by removing the TiO2 nanotube (NT) layers prepared by an anodization process. To determine the in vitro effects of the ENF surface, cell proliferation assay, alkaline phosphatase activity assay, alizarin red staining, western blotting, and immunocytochemistry were performed. Atomic force microscopy and scanning electron microscopy analysis show that the ENF surface has an ultrafine surface roughness with highly aligned nanoporous morphology. hMSCs on ENF surfaces exhibit increased proliferation and enhanced osteogenic differentiation as compared to the ordered TiO2 nanotubular and compact TiO2 surfaces. Surface modification with the ENF process is a promising technique for fabricating osteointegrative implant materials with a highly bioactive, rigid and purified nano surfaces.


Assuntos
Diferenciação Celular , Eletroquímica , Células-Tronco Mesenquimais/citologia , Nanotecnologia , Osteogênese , Titânio/farmacologia , Fosfatase Alcalina/metabolismo , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Nanotubos/química , Nanotubos/ultraestrutura , Osteocalcina/metabolismo , Osteogênese/efeitos dos fármacos , Propriedades de Superfície
14.
Mol Brain ; 12(1): 23, 2019 03 25.
Artigo em Inglês | MEDLINE | ID: mdl-30909971

RESUMO

It is now generally accepted that the extra-skeleton functionalities of bone are multifaceted. Its endocrine functions came first to light when it was realized that osteoblasts, the bone forming cells, maintain energy homeostasis by improving glucose metabolism, insulin sensitivity and energy expenditure through osteocalcin, a multipurpose osteokine secreted by osteoblasts. Recently, the emerging knowledge on the functional aspects of this osteokine expanded to properties including adult and maternal regulation of cognitive functions. Therapeutic potential of this osteokine has also been recently reported in experimental Parkinson's disease models. This review highlights such findings on the functions of osteocalcin in the brain and emphasizes on exploring and analyzing much more in-depth basic and clinical studies.


Assuntos
Encéfalo/metabolismo , Cognição/fisiologia , Doença dos Neurônios Motores/metabolismo , Doença dos Neurônios Motores/fisiopatologia , Osteocalcina/metabolismo , Transdução de Sinais , Animais , Humanos , Doença dos Neurônios Motores/terapia , Fármacos Neuroprotetores/metabolismo
15.
Mater Sci Eng C Mater Biol Appl ; 99: 710-718, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30889744

RESUMO

Surface chemistry and topography can determinatively affect the osseointegration of dental implants. Strontium (Sr) has a significant effect on the promotion of bone formation and inhibitation of bone resorption. The emphasis of this study lies on the evaluation of a new surface treatment that aims to improve the early osseointegration of dental implantation both in vitro and in vivo. A hydrothermal method was used to prepare an SrTiO3 incorporation on sandblasted large-grit double acid-etched (SLA) titanium surfaces in SrCl2 solution. The composition and morphology of the SrTiO3 doped surface were analyzed by X-ray diffraction, X-ray photoelectron spectroscopy,and scanning electron microscopy. In addition, the external release figure of Sr was examined by inductively coupled plasma mass spectrometry. The proliferation, adhesion and differentiation of MC3T3-E1 cells on this surface were evaluated in vitro and presented a significant increase in SLA-Sr group compared with that in SLA group. An in vivo study in 24 New Zealand rabbits indicated a remarkable growth in the volume of direct bone-to-implant contact and peri-implant bone in SLA-Sr group, which were compared with SLA group after 3 and 6 weeks, and removal torque tests exhibited a higher torque removal value of SLA-Sr implants. The study gave the result that the biological effect of SLA-Sr implants was significantly superior to that of the SLA implants at the early stage of osseointegration.


Assuntos
Osteogênese/efeitos dos fármacos , Óxidos/farmacologia , Estrôncio/farmacologia , Titânio/farmacologia , Ataque Ácido Dentário , Fosfatase Alcalina/metabolismo , Animais , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Íons , Masculino , Camundongos , Nanoestruturas/química , Osteocalcina/metabolismo , Espectroscopia Fotoeletrônica , Coelhos , Estrôncio/química , Propriedades de Superfície , Torque , Difração de Raios X
16.
Med Sci Monit ; 25: 2289-2295, 2019 Mar 29.
Artigo em Inglês | MEDLINE | ID: mdl-30923307

RESUMO

BACKGROUND The essence of osteoporosis is mainly the imbalance of bone formation and absorption. Previous studies indicated that SIRT1 is closely related to bone metabolism and bone mass as a regulator of bone mass. The literature reports that microRNAs are significant regulators of osteoblast proliferation and differentiation. MATERIAL AND METHODS In this study, SIRT1 protein and mRNA levels were examined by Western blot and RT-PCR. Osteogenic proliferation was examined by CCK8 assay and osteogenic markers, including ALP, OCN, and RUNX2, were examined by ELISA. The target of miR-132-3p was identified by luciferase reporter assay. RESULTS LPS downregulated the SIRT1 protein level and ß-glycerophosphate upregulated the SIRT1 protein level. The results demonstrated that SIRT1 overexpression promoted the proliferation and differentiation in MC3T3-E1 cells, and SIRT1 interference had the opposite effect. Luciferase reporter assay revealed that miR-132-3p inhibited the reporter gene activity of SIRT1. LPS upregulated the mRNA level of miR-132-3p, and ß-glycerophosphate downregulated the mRNA level of miR-132-3p. CONCLUSIONS miR-132-3p is a pivotal regulator in osteogenic proliferation and differentiation by targeting SIRT1.


Assuntos
MicroRNAs/genética , Osteoporose/genética , Sirtuína 1/metabolismo , Células 3T3 , Fosfatase Alcalina/metabolismo , Animais , Diferenciação Celular/genética , Linhagem Celular , Proliferação de Células/genética , Células Cultivadas , China , Regulação da Expressão Gênica/genética , Lipopolissacarídeos/farmacologia , Camundongos , MicroRNAs/fisiologia , Osteoblastos/metabolismo , Osteocalcina/metabolismo , Osteogênese/efeitos dos fármacos , Osteoporose/metabolismo , Sirtuína 1/genética
17.
Mater Sci Eng C Mater Biol Appl ; 99: 905-918, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30889765

RESUMO

BACKGROUND: Due to unmet need for bone augmentation, our aim was to promote osteogenic differentiation of human adipose stem cells (hASCs) encapsulated in gellan gum (GG) or collagen type I (COL) hydrogels with bioactive glass (experimental glass 2-06 of composition [wt-%]: Na2O 12.1, K2O 14.0, CaO 19.8, P2O5 2.5, B2O3 1.6, SiO2 50.0) extract based osteogenic medium (BaG OM) for bone construct development. GG hydrogels were crosslinked with spermidine (GG-SPD) or BaG extract (GG-BaG). METHODS: Mechanical properties of cell-free GG-SPD, GG-BaG, and COL hydrogels were tested in osteogenic medium (OM) or BaG OM at 0, 14, and 21 d. Hydrogel embedded hASCs were cultured in OM or BaG OM for 3, 14, and 21 d, and analyzed for viability, cell number, osteogenic gene expression, osteocalcin production, and mineralization. Hydroxyapatite-stained GG-SPD samples were imaged with Optical Projection Tomography (OPT) and Selective Plane Illumination Microscopy (SPIM) in OM and BaG OM at 21 d. Furthermore, Raman spectroscopy was used to study the calcium phosphate (CaP) content of hASC-secreted ECM in GG-SPD, GG-BaG, and COL at 21 d in BaG OM. RESULTS: The results showed viable rounded cells in GG whereas hASCs were elongated in COL. Importantly, BaG OM induced significantly higher cell number and higher osteogenic gene expression in COL. In both hydrogels, BaG OM induced strong mineralization confirmed as CaP by Raman spectroscopy and significantly improved mechanical properties. GG-BaG hydrogels rescued hASC mineralization in OM. OPT and SPIM showed homogeneous 3D cell distribution with strong mineralization in BaG OM. Also, strong osteocalcin production was visible in COL. CONCLUSIONS: Overall, we showed efficacious osteogenesis of hASCs in 3D hydrogels with BaG OM with potential for bone-like grafts.


Assuntos
Tecido Adiposo/citologia , Diferenciação Celular , Colágeno Tipo I/farmacologia , Vidro/química , Osteogênese , Polissacarídeos Bacterianos/farmacologia , Células-Tronco/citologia , Animais , Biomarcadores/metabolismo , Calcificação Fisiológica/efeitos dos fármacos , Contagem de Células , Diferenciação Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Força Compressiva , Reagentes para Ligações Cruzadas/química , Durapatita/química , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Íons , Pessoa de Meia-Idade , Minerais/química , Osteocalcina/metabolismo , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Ratos , Soro/metabolismo , Análise Espectral Raman , Células-Tronco/efeitos dos fármacos , Tecidos Suporte/química
18.
Expert Opin Investig Drugs ; 28(5): 489-496, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30908082

RESUMO

BACKGROUND: An efficacious anti-inflammatory corticosteroid with reduced side effects has been long sought. We report the pooled results from three clinical proof-of-mechanism Phase I studies of BI 653048 in healthy subjects, a functionally selective, nonsteroidal glucocorticoid (GC). RESEARCH DESIGN AND METHODS: Three Phase I trials were conducted: a single rising-dose study and a multiple rising-dose study to evaluate the safety, tolerability, and pharmacokinetics of BI 653048, and a multiple parallel-arm-dose study with intravenous lipopolysaccharide challenge to assess in vivo pharmacodynamics. The pharmacodynamics, efficacy, and safety of BI 653048 and prednisolone were compared. RESULTS: Treatment with 200 mg BI 653048 was associated with a reduced expression of IL1R2, ITGB3, and SDPR versus 20 mg prednisolone; comparable levels of FKBP5, ZBTB16, and DDIT4 expression were observed. Changes in C-peptide, glucose, insulin, and cortisol were moderate compared with prednisolone. A greater reduction of osteocalcin was observed with 200 mg BI 653048 versus 20 mg prednisolone. Comparable anti-inflammatory efficacy was demonstrated for 200 mg BI 653048 and 20 mg prednisolone. BI 653048 was well tolerated in healthy subjects. CONCLUSION: BI 653048 demonstrated the desired anti-inflammatory effects of the nonsteroidal GC; however, the undesirable side-effect profile associated with GC steroids could not be disassociated from BI 653048. TRIAL REGISTRATION: ClinicalTrials.gov identifiers NCT02217644, NCT02217631, and NCT02224105.


Assuntos
Anti-Inflamatórios/farmacologia , Benzamidas/farmacologia , Glucocorticoides/farmacologia , Piridinas/farmacologia , Pirróis/farmacologia , Receptores de Glucocorticoides/agonistas , Adulto , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/efeitos adversos , Benzamidas/administração & dosagem , Benzamidas/efeitos adversos , Ensaios Clínicos Fase I como Assunto , Glucocorticoides/administração & dosagem , Glucocorticoides/efeitos adversos , Humanos , Inflamação/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Osteocalcina/metabolismo , Prednisolona/efeitos adversos , Prednisolona/farmacologia , Piridinas/administração & dosagem , Piridinas/efeitos adversos , Pirróis/administração & dosagem , Pirróis/efeitos adversos , Ensaios Clínicos Controlados Aleatórios como Assunto , Adulto Jovem
19.
Int J Mol Sci ; 20(6)2019 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-30917596

RESUMO

Cadmium is a common environmental pollutant that causes bone damage. However, the effects of cadmium on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs) and its mechanism of action in this process are unclear. Here, we determined the effects of cadmium chloride (CdCl2) on the osteogenic differentiation of BMMSCs and the potential mechanism involved in this process. As determined in the present investigation, CdCl2, in a concentration-dependent manner, affected the viability of BMMSCs and their cytoskeletons. Exposure to 0.1 or 0.2 µM CdCl2 inhibited osteogenic differentiation of BMMSCs, which was reflected in the down-regulation of osteoblast-related genes (ALP, OCN, Runx2, OSX, and OPN); in suppression of the protein expression of alkaline phosphatase (ALP) and runt-related transcription factor 2 (Runx2); and in decreased ALP activity and capacity for mineralization. Moreover, mRNA microarray was performed to determine the roles of these factors in BMMSCs treated with CdCl2 in comparison to control BMMSCs. As determined with the microarrays, the Wingless-type (Wnt), mothers against decapentaplegic and the C. elegans gene Sam (SMAD), and Janus kinase-Signal Transducers and Activators of Transcription (JAK-STAT) signaling pathways were involved in the effects caused by CdCl2. Moreover, during differentiation, the protein levels of Wnt3a, ß-catenin, lymphoid enhancer factor 1 (LEF1), and T-cell factor 1 (TCF1) were reduced by CdCl2. The current research shows that CdCl2 suppresses the osteogenesis of BMMSCs via inhibiting the Wnt/ß-catenin pathway. The results establish a previously unknown mechanism for bone injury induced by CdCl2.


Assuntos
Células da Medula Óssea/metabolismo , Cloreto de Cádmio/farmacologia , Diferenciação Celular , Células-Tronco Mesenquimais/metabolismo , Osteoblastos/metabolismo , Via de Sinalização Wnt , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/efeitos dos fármacos , Células Cultivadas , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteocalcina/genética , Osteocalcina/metabolismo , Osteopontina/genética , Osteopontina/metabolismo , Ratos , Ratos Sprague-Dawley , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
20.
Cell Tissue Bank ; 20(1): 61-75, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30729369

RESUMO

To investigate the impact of different anticoagulants and coagulants with autologous platelet-rich plasma (PRP) in order to evaluate the clinical application of PRP standardization. Bone marrow stem cells (BMSCs) were seeded into autologous PRP gel scaffolds with different anticoagulants (EDTA, heparin sodium HS, and sodium citrate SC) as well as control group (the whole blood group). Quality of PRP was evaluated and flow cytometric assay was used to detect the activity of the platelet (CD62p, PAC-1). BMSCs were also seeded into PRP with different coagulants (Thrombin, Collagen-I, ADP) as well as PRP un-activated (negative group) and L-DMEM complete culture without PRP (control group). The effects of different coagulants with PRP on proliferation, osteogenic differentiation of BMSCs were analyzed by methyl thiazolyl tetrazolium assay (MTT), ALP staining, Von Kossa staining, Confocal microscopic observation, RT-PCR and Western Blot at the morphological, cellular and molecular levels. Different anticoagulants (EDTA, HS, and SC) could affect the quality of PRP. EDTA group revealed the best quality and activity (CD62p, PAC-1). With different coagulants (Thrombin, Collagen-I and ADP) in the proliferation of BMSCs, the MTT assay showed that the proliferation of BMSCs was increased in all groups with time. On the sixth day of culture, the cell number of each PRP group was significantly higher than that in the control group (P < 0.05), while the most rapidly increasing was found in Collagen-I group. The cumulative release of growth factor (TGF-ß1, PDGF) at each time point in the PRP gel of the four groups was higher than that in the control group (P < 0.05). Collagen-I was considered as the best PRP coagulant. When thrombin was used as a platelet coagulant, the release of growth factor in PRP was rapid and direct, while the release of growth factor in Collagen-I-activated PRP was sustained and slow, and the total release of ADP-activated PRP growth factors was the lowest. The study demonstrated the similar outcome in osteogenic differentiation. In terms of gene expression and western bolt, the PCR results showed that the expression levels of OCN gene and RUNX2 protein in each PRP group were higher than that in the control group (P < 0.05). Different anticoagulants caused different degrees of lysis and spontaneous activation of platelets, which lead to different quality of PRP. Compared with HS and SC, EDTA could maintain the structural integrity of platelets, reduce their spontaneous activation, and increase the release of PRP growth factors for a longer period of time, thus ensuring the biomass of PRP. In addition, different coagulants also showed different results in the proliferation as well as osteogenic differentiation of BMSCs. Compared with Thrombin and ADP, Collagen-I may be a better choice.


Assuntos
Anticoagulantes/farmacologia , Coagulantes/farmacologia , Plasma Rico em Plaquetas/metabolismo , Animais , Bioensaio , Plaquetas/citologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Linhagem da Célula/efeitos dos fármacos , Linhagem da Célula/genética , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/genética , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/genética , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Fosfatase 2 de Especificidade Dupla/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Osteocalcina/genética , Osteocalcina/metabolismo , Osteogênese/efeitos dos fármacos , Osteogênese/genética , Selectina-P/metabolismo , Fator de Crescimento Derivado de Plaquetas/farmacologia , Coelhos , Padrões de Referência , Fator de Crescimento Transformador beta1/farmacologia
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