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1.
Molecules ; 28(8)2023 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-37110788

RESUMO

In this study, we aimed to develop natural and/or functional materials with antioxidant and anti-inflammatory effects. We obtained extracts from natural plants through an oil and hot-water extraction process and prepared an extract composite of an effective unsaturated fatty acid complex (EUFOC). Furthermore, the antioxidant effect of the extract complex was evaluated, and the anti-inflammatory effect was explored by assessing its inhibitory effect on nitric oxide production through its HA-promoting effect. We conducted a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay to evaluate the cell viability of the EUFOC, and the results showed that EUFOC was not cytotoxic at the test concentrations. In addition, it showed no endogenous cytotoxicity in HaCaT (human keratinocyte) cells. The EUFOC showed excellent 1,1-diphenyl-2-picrylhydrazyl- and superoxide-scavenging abilities. Moreover, it exerted an inhibitory effect on NO production at concentrations that did not inhibit cell viability. The secretion of all the cytokines was increased by lipopolysaccharide (LPS) treatment; however, this was inhibited by the EUFOC in a concentration-dependent manner. In addition, hyaluronic acid content was markedly increased by the EUFOC in a dose-dependent manner. These results suggest that the EUFOC has excellent anti-inflammatory and antioxidant properties, and hence, it can be used as a functional material in various fields.


Assuntos
Antioxidantes , Ácido Hialurônico , Humanos , Antioxidantes/farmacologia , Extratos Vegetais/farmacologia , Óxido Nítrico/metabolismo , Anti-Inflamatórios/farmacologia , Citocinas
2.
J Cell Mol Med ; 26(16): 4520-4529, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35796406

RESUMO

Scoparone (SCOP), an active and efficient coumarin compound derived from Artemisia capillaris Thunb, has been used as a traditional Chinese herbal medicine. Herein, we investigated the effects of SCOP on the osteogenic processes using MC3T3-E1 pre-osteoblasts in in vitro cell systems. SCOP (C11 H10 O4 , > 99.17%) was purified and identified from A. capillaries. SCOP (0.1 to 100 µM concentrations) did not have cytotoxic effects in pre-osteoblasts; however, it promoted alkaline phosphatase (ALP) staining and activity, and mineralized nodule formation under early and late osteogenic induction. SCOP elevated osteogenic signals through the bone morphogenetic protein 2 (BMP2)-Smad1/5/8 pathway, leading to the increased expression of runt-related transcription factor 2 (RUNX2) with its target protein, matrix metallopeptidase 13 (MMP13). SCOP also induced the non-canonical BMP2-MAPKs pathway, but not the Wnt3a-ß-catenin pathway. Moreover, SCOP promoted autophagy, migration and adhesion under the osteogenic induction. Overall, the findings of this study demonstrated that SCOP has osteogenic effects associated with cell differentiation, adhesion, migration, autophagy and mineralization.


Assuntos
Subunidade alfa 1 de Fator de Ligação ao Core , Osteogênese , Autofagia , Proteína Morfogenética Óssea 2/metabolismo , Diferenciação Celular , Linhagem Celular , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Cumarínicos/farmacologia , Osteoblastos/metabolismo
3.
Pharmacol Res ; 184: 106423, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-36064078

RESUMO

BMP2 is clinically used as an ectopic bone inducer and plays a significant role in bone development, formation, and diseases. Chitinase 3-like 1 protein (Chi3L1) is found in the skeletal system. However, Chi3L1-mediated bone metabolism and aging-related bone erosion via BMP2 signaling have not yet been demonstrated. Herein, Chi3L1 increased BMP2-induced osteoblast differentiation in mesenchymal precursor cells and human primary osteoblasts. Chi3L1KO(-/-) showed abnormal bone development, and primary osteoblasts isolated from Chi3L1KO(-/-) exhibited impaired osteoblast differentiation and maturation. Chi3L1 also potentiated BMP2 signaling and RUNX2 expression in primary osteoblasts. Chi3L1 interacted with BMPRIa, which increased the surface expression of BMPRIa and promoted BMP2 signaling to induce osteoblast differentiation. Chi3L1KO(-/-) mice showed bone formation reduced with a decrease in RUNX2 expression in calvarial defects. Chi3L1KO(-/-) mice exhibited aging-related osteoporotic bone loss with decreases in the levels of RUNX2 and OPG, while serum PYD level and osteoclast number increased. Chi3L1 increased OPG via non-canonical BMP2 signaling in osteoblasts, which suppressed osteoclastogenesis in BMMs. Furthermore, ROC analysis showed that serum Chi3L1 level clinically decreased in osteoporosis patients. Our findings demonstrate that Chi3L1 promotes bone formation, suppresses osteoclastogenesis, and prevents aging-related osteoporosis.


Assuntos
Quitinases , Osteoporose , Animais , Biomarcadores/metabolismo , Diferenciação Celular , Proteína 1 Semelhante à Quitinase-3/genética , Proteína 1 Semelhante à Quitinase-3/metabolismo , Quitinases/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Humanos , Camundongos , Osteoblastos/metabolismo , Osteogênese , Osteoporose/tratamento farmacológico , Osteoporose/metabolismo
4.
Sensors (Basel) ; 22(6)2022 Mar 10.
Artigo em Inglês | MEDLINE | ID: mdl-35336325

RESUMO

Heart rate variability (HRV) is closely related to changes in the autonomic nervous system (ANS) associated with stress and pain. In this study, we investigated whether HRV could be used to assess cancer pain in mice with peritoneal metastases. At 12 days after cancer induction, positive indicators of pain such as physiological characteristics, appearance, posture, and activity were observed, and time- and frequency-domain HRV parameters such as mean R-R interval, square root of the mean squared differences of successive R-R intervals, and percentage of successive R-R interval differences greater than 5 ms, low frequency (LF), high frequency (HF), and ratio of LF and HF power, were found to be significantly decreased. These parameters returned to normal after analgesic administration. Our results indicate that overall ANS activity was decreased by cancer pain and that HRV could be a useful tool for assessing pain.


Assuntos
Dor do Câncer , Neoplasias Peritoneais , Animais , Sistema Nervoso Autônomo , Frequência Cardíaca/fisiologia , Camundongos
5.
Int J Mol Sci ; 23(15)2022 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-35955423

RESUMO

Triterpenes are a diverse group of natural compounds found in plants. Soyasapogenol B (SoyB) from Arachis hypogaea (peanut) has various pharmacological properties. This study aimed to elucidate the pharmacological properties and mechanisms of SoyB in bone-forming cells. In the present study, 1-20 µM of SoyB showed no cell proliferation effects, whereas 30-100 µM of SoyB increased cell proliferation in MC3T3-E1 cells. Next, osteoblast differentiation was analyzed, and it was found that SoyB enhanced ALP staining and activity and bone mineralization. SoyB also induced RUNX2 expression in the nucleus with the increased phosphorylation of Smad1/5/8 and JNK2 during osteoblast differentiation. In addition, SoyB-mediated osteoblast differentiation was not associated with autophagy and necroptosis. Furthermore, SoyB increased the rate of cell migration and adhesion with the upregulation of MMP13 levels during osteoblast differentiation. The findings of this study provide new evidence that SoyB possesses biological effects in bone-forming cells and suggest a potentially beneficial role for peanut-based foods.


Assuntos
Arachis , Triterpenos , Autofagia , Diferenciação Celular , Linhagem Celular , Necroptose , Ácido Oleanólico/análogos & derivados , Osteoblastos/metabolismo , Saponinas , Triterpenos/metabolismo , Triterpenos/farmacologia
6.
Int J Mol Sci ; 22(21)2021 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-34768792

RESUMO

Stem cells have received attention in various diseases, such as inflammatory, cancer, and bone diseases. Mesenchymal stem cells (MSCs) are multipotent stem cells that are critical for forming and repairing bone tissues. Herein, we isolated calycosin-7-O-ß-glucoside (Caly) from the roots of Astragalus membranaceus, which is one of the most famous medicinal herbs, and investigated the osteogenic activities of Caly in MSCs. Caly did not affect cytotoxicity against MSCs, whereas Caly enhanced cell migration during the osteogenesis of MSCs. Caly increased the expression and enzymatic activities of ALP and the formation of mineralized nodules during the osteogenesis of MSCs. The osteogenesis and bone-forming activities of Caly are mediated by bone morphogenetic protein 2 (BMP2), phospho-Smad1/5/8, Wnt3a, phospho-GSK3ß, and phospho-AKT, inducing the expression of runt-related transcription factor 2 (RUNX2). In addition, Caly-mediated osteogenesis and RUNX2 expression were attenuated by noggin and wortmannin. Moreover, the effects were validated in pre-osteoblasts committed to the osteoblast lineages from MSCs. Overall, our results provide novel evidence that Caly stimulates osteoblast lineage commitment of MSCs by triggering RUNX2 expression, suggesting Caly as a potential anabolic drug to prevent bone diseases.


Assuntos
Calcificação Fisiológica/efeitos dos fármacos , Glucosídeos/farmacologia , Isoflavonas/farmacologia , Osteogênese/efeitos dos fármacos , Animais , Astragalus propinquus/metabolismo , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Proteína Morfogenética Óssea 2/metabolismo , Calcificação Fisiológica/fisiologia , Diferenciação Celular/efeitos dos fármacos , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Glucosídeos/isolamento & purificação , Glucosídeos/metabolismo , Humanos , Isoflavonas/isolamento & purificação , Isoflavonas/metabolismo , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/fisiologia , Camundongos , Células NIH 3T3 , Osteoblastos/metabolismo , Osteogênese/fisiologia , Extratos Vegetais/isolamento & purificação , Extratos Vegetais/farmacologia
7.
Nanomedicine ; 24: 102129, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31760162

RESUMO

Osseointegration is important in osteopenia and osteoporosis patients due to their low bone densities. Gold nanoparticles (GNPs) are greatly beneficial materials as osteogenic agents. The aim of this study is to investigate osseointegration between bones and double layers of GNP-immobilized titanium (Ti) implants. The physicochemical properties of the Ti surface were evaluated by scanning electron microscopy, by atomic force microscopy, by means of the contact angle using water drops, and by x-ray photoelectron spectroscopy. Osteogenic differentiation of human bone-marrow-derived mesenchymal stem cells was analyzed and showed the higher values in double layers of GNP (GNP2) groups. In addition, we performed an in vivo study using hydroxyapatite (HA) and GNP2 spine pedicle screws in ovariectomized (OVX) and SHAM rabbits. Osseointegration parameters also showed higher values in GNP2 than in HA groups. These findings suggest that implants with double layers of GNPs can be a useful alternative in osteoporotic patients.


Assuntos
Durapatita/química , Ouro/química , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/metabolismo , Nanopartículas Metálicas/química , Osseointegração/efeitos dos fármacos , Titânio/química , Titânio/farmacologia , Fosfatase Alcalina/metabolismo , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Proliferação de Células/efeitos dos fármacos , Proliferação de Células/fisiologia , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Coelhos , Reação em Cadeia da Polimerase Via Transcriptase Reversa
8.
Int J Mol Sci ; 21(4)2020 Feb 24.
Artigo em Inglês | MEDLINE | ID: mdl-32102392

RESUMO

Exosomes are nanosized vesicles (30-140 nm) of endocytic origin that play important roles in regenerative medicine. They are derived from cell membranes during endocytic internalization and stabilize in biological fluids such as blood and synovia. Temporomandibular joint osteoarthritis (TMJ OA) is a degenerative disease, which, in addition to chronic pain, is characterized by progressive cartilage breakdown, condylar bone remodeling, and synovitis. However, traditional clinical treatments have limited symptom- and structure-modifying effects to restore damaged cartilage and other TMJ tissues. This is due to the limited self-healing capacity of condylar cartilage. Recently, stem-cell-derived exosomes have been studied as an alternative therapeutic approach to tissue repair and regeneration. It is known that trophic regulation of mesenchymal stem cells (MSCs) has anti-inflammatory and immunomodulatory effects under pathological conditions, and research on MSC-derived exosomes is rapidly accumulating. MSC-derived exosomes mimic the major therapeutic effects of MSCs. They affect the activity of immune effector cells and possess multilineage differentiation potential, including chondrogenic and osteogenic differentiation. Furthermore, exosomes are capable of regenerating cartilage or osseous compartments and restoring injured tissues and can treat dysfunction and pain caused by TMJ OA. In this review, we looked at the uniqueness of TMJ, the pathogenesis of TMJ OA, and the potential role of MSC-derived exosomes for TMJ cartilage and bone regeneration.


Assuntos
Exossomos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Osteoartrite/metabolismo , Regeneração , Medicina Regenerativa/métodos , Articulação Temporomandibular/metabolismo , Animais , Humanos , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Osteoartrite/fisiopatologia , Osteogênese , Articulação Temporomandibular/patologia , Articulação Temporomandibular/fisiopatologia
9.
J Mater Sci Mater Med ; 30(6): 63, 2019 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-31127379

RESUMO

The menisci have crucial roles in the knee, chondroprotection being the primary. Meniscus repair or substitution is favored in the clinical management of the meniscus lesions with given indications. The outstanding challenges with the meniscal scaffolds include the required biomechanical behavior and features. Suturability is one of the prerequisites for both implantation and implant survival. Therefore, we proposed herein a novel highly interconnected suturable porous scaffolds from regenerated silk fibroin that is reinforced with 3D-printed polycaprolactone (PCL) mesh in the middle, on the transverse plane to enhance the suture-holding capacity. Results showed that the reinforcement of the silk fibroin scaffolds with the PCL mesh increased the suture retention strength up to 400%, with a decrease in the mean porosity and an increase in crystallinity from 51.9 to 55.6%. The wet compression modulus values were significantly different for silk fibroin, and silk fibroin + PCL mesh by being 0.16 ± 0.02, and 0.40 ± 0.06 MPa, respectively. Both scaffolds had excellent interconnectivity (>99%), and a high water uptake feature (>500%). The tissue's infiltration and formation of new blood vessels were assessed by means of performing an in vivo subcutaneous implantation of the silk fibroin + PCL mesh scaffolds that were seeded with primary human meniscocytes or stem cells. Regarding suturability and in vivo biocompatibility, the findings of this study indicate that the silk fibroin + PCL mesh scaffolds are suitable for further studies to be carried out for meniscus tissue engineering applications such as the studies involving orthotopic meniscal models and fabrication of patient-specific implants.


Assuntos
Materiais Biocompatíveis/química , Fibroínas/química , Poliésteres/química , Impressão Tridimensional , Telas Cirúrgicas , Animais , Bombyx , Força Compressiva , Humanos , Menisco/citologia , Microscopia Eletrônica de Varredura , Porosidade , Pressão , Regeneração , Células-Tronco/citologia , Estresse Mecânico , Suturas , Engenharia Tecidual/métodos , Alicerces Teciduais , Água/química , Microtomografia por Raio-X
10.
Sci Technol Adv Mater ; 20(1): 826-836, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31489055

RESUMO

In an aging society, bone disorders such as osteopenia, osteoporosis, and degenerative arthritis cause serious public health problems. In order to solve these problems, researchers continue to develop therapeutic agents, increase the efficacy of developed therapeutic agents, and reduce side effects. Gold nanoparticles (GNPs) are widely used in tissue engineering applications as biosensors, drug delivery carriers, and bioactive materials. Their special surface property enables easy conjugation with ligands including functional groups such as thiols, phosphines, and amines. This creates an attractive advantage to GNPs for use in the bone tissue engineering field. However, GNPs alone are limited in their biological effects. In this study, we used thiol-PEG-vitamin D (SPVD) to conjugate vitamin D, an essential nutrient critical for maintaining normal skeletal homeostasis, to GNPs. To characterize vitamin D-conjugated GNPs (VGNPs), field emission transmission electron microscopy, energy dispersive X-ray spectroscopy, dynamic light scattering, and ultraviolet/visible absorption analysis were carried out. The developed VGNPs were well bound through the thiol groups between GNPs and vitamin D, and were fabricated in size of 60 nm. Moreover, to demonstrate VGNPs osteogenic differentiation effect, various assays were carried out through cell viability test, alkaline phosphatase assay, calcium deposition assay, real-time polymerase chain reaction, and immunofluorescence staining. As a result, the fabricated VGNPs were found to effectively enhance osteogenic differentiation of human adipose-derived stem cells (hADSCs) in vitro. Based on these results, VGNPs can be utilized as functional nanomaterials for bone regeneration in the tissue engineering field.

11.
Biochem Biophys Res Commun ; 496(3): 785-791, 2018 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-29395078

RESUMO

An agarose scaffold can be useful for supporting and guiding injured axons after spinal cord injury (SCI), but the electrophysiological signal of regenerated axon in scaffolds has not yet been determined. The current study investigated whether a Matrigel-loaded agarose scaffold would enhance the regeneration of axons after SCI. Moreover, the functional connectivity of regenerated axons within the channels of the scaffold was evaluated by directly recording motor evoked potentials. Our data showed that the agarose scaffold containing Matrigel can support and enhance linearly organized axon regeneration after SCI. Additionally, motor evoked potentials were successfully recorded from regenerated axons. These results demonstrate that an agarose scaffold loaded with Matrigel could promote the regeneration of axons and guide the reconnection of functional axons after SCI.


Assuntos
Axônios/patologia , Colágeno/química , Regeneração Tecidual Guiada/instrumentação , Laminina/química , Regeneração Nervosa/fisiologia , Proteoglicanas/química , Traumatismos da Medula Espinal/patologia , Traumatismos da Medula Espinal/terapia , Alicerces Teciduais , Animais , Materiais Biomiméticos/síntese química , Combinação de Medicamentos , Desenho de Equipamento , Análise de Falha de Equipamento , Masculino , Crescimento Neuronal , Próteses e Implantes , Ratos , Ratos Sprague-Dawley , Recuperação de Função Fisiológica , Sefarose/química , Traumatismos da Medula Espinal/fisiopatologia , Resultado do Tratamento
12.
Bioelectromagnetics ; 39(5): 394-404, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29709064

RESUMO

Although we recently demonstrated that static magnetic fields (SMFs) of 3, 15, and 50 mT stimulate osteoblastic differentiation, the effects of SMFs on osteoclastogenesis are still poorly understood. This study focused on the suppressive effects of SMFs on receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastogenesis and bone resorption. Direct SMFs inhibit RANKL-induced multinucleated osteoclast formation, tartrate-resistant acid phosphatase activity, and bone resorption in mouse bone marrow-derived macrophage cells. The conditioned medium from osteoblasts treated with SMFs also resulted in the inhibition of osteoclast differentiation as well as resorption. The RANKL-induced expression of osteoclast-specific transcription factors, such as c-Fos and NFATc1, was remarkably downregulated by SMF at 15 mT. In addition, SMF inhibited RANKL-activated Akt, glycogen synthase kinase 3ß (GSK3ß), extracellular signal-regulated kinase, c-jun N-terminal protein kinase, mitogen-activated protein kinase (MAPK), and nuclear factor-κB (NF-κB) formation. These findings indicate that SMF-mediated attenuation of RANKL-induced Akt, GSK3ß, MAPK, and NF-κB pathways could contribute to the direct and indirect inhibition of osteoclast formation and bone resorption. Therefore, SMFs could be developed as a therapeutic agent against periprosthetic or peri-implant osteolysis. Additionally, these could be used against osteolytic diseases such as osteoporosis and rheumatoid arthritis. Bioelectromagnetics. 39:394-404, 2018. © 2018 Wiley Periodicals, Inc.


Assuntos
Células da Medula Óssea/fisiologia , Diferenciação Celular/fisiologia , Campos Magnéticos , Osteoclastos/fisiologia , Animais , Células da Medula Óssea/citologia , Reabsorção Óssea/patologia , Reabsorção Óssea/fisiopatologia , Células Cultivadas , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Feminino , Glicogênio Sintase Quinase 3 beta/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Camundongos Endogâmicos ICR , NF-kappa B/metabolismo , Fatores de Transcrição NFATC/metabolismo , Osteoclastos/citologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas Proto-Oncogênicas c-fos/metabolismo , Ligante RANK/metabolismo , Transdução de Sinais
13.
Adv Exp Med Biol ; 1078: 103-117, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30357620

RESUMO

Tissues are often damaged by physical trauma, infection or tumors. A slight injury heals naturally through the normal healing process, while severe injury causes serious health implications. Therefore, many efforts have been devoted to treat and repair various tissue defects. Recently, tissue engineering approaches have attracted a rapidly growing interest in biomedical fields to promote and enhance healing and regeneration of large-scale tissue defects. On the other hand, with the recent advances in nanoscience and nanotechnology, various nanomaterials have been suggested as novel biomaterials. Graphene, a two-dimensional atomic layer of graphite, and its derivatives have recently been found to possess promoting effects on various types of cells. In addition, their unique properties, such as outstanding mechanical and biological properties, allow them to be a promising option for hard tissue regeneration. Herein, we summarized recent research advances in graphene-based nanocomposites for hard tissue regeneration, and highlighted their promising potentials in biomedical and tissue engineering.


Assuntos
Regeneração Óssea , Grafite , Nanocompostos , Engenharia Tecidual , Materiais Biocompatíveis , Humanos , Nanotecnologia
14.
Int J Mol Sci ; 19(6)2018 06 12.
Artigo em Inglês | MEDLINE | ID: mdl-29895782

RESUMO

The present study evaluated the protective effects of melatonin in ethanol (EtOH)-induced senescence and osteoclastic differentiation in human periodontal ligament cells (HPDLCs) and cementoblasts and the underlying mechanism. EtOH increased senescence activity, levels of reactive oxygen species (ROS) and the expression of cell cycle regulators (p53, p21 and p16) and senescence-associated secretory phenotype (SASP) genes (interleukin [IL]-1ß, IL-6, IL-8 and tumor necrosis factor-α) in HPDLCs and cementoblasts. Melatonin inhibited EtOH-induced senescence and the production of ROS as well as the increased expression of cell cycle regulators and SASP genes. However, it recovered EtOH-suppressed osteoblastic/cementoblastic differentiation, as evidenced by alkaline phosphatase activity, alizarin staining and mRNA expression levels of Runt-related transcription factor 2 (Runx2) and osteoblastic and cementoblastic markers (glucose transporter 1 and cementum-derived protein-32) in HPDLCs and cementoblasts. Moreover, it inhibited EtOH-induced osteoclastic differentiation in mouse bone marrow⁻derived macrophages (BMMs). Inhibition of protein never in mitosis gene A interacting-1 (PIN1) by juglone or small interfering RNA reversed the effects of melatonin on EtOH-mediated senescence as well as osteoblastic and osteoclastic differentiation. Melatonin blocked EtOH-induced activation of mammalian target of rapamycin (mTOR), AMP-activated protein kinase (AMPK), mitogen-activated protein kinase (MAPK) and Nuclear factor of activated T-cells (NFAT) c-1 pathways, which was reversed by inhibition of PIN1. This is the first study to show the protective effects of melatonin on senescence-like phenotypes and osteoclastic differentiation induced by oxidative stress in HPDLCs and cementoblasts through the PIN1 pathway.


Assuntos
Cemento Dentário/citologia , Etanol/farmacologia , Melatonina/farmacologia , Osteoclastos/efeitos dos fármacos , Osteoclastos/metabolismo , Ligamento Periodontal/citologia , Diferenciação Celular/efeitos dos fármacos , Células Cultivadas , Senescência Celular/efeitos dos fármacos , Cemento Dentário/metabolismo , Humanos , Peptidilprolil Isomerase de Interação com NIMA/metabolismo , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteoclastos/citologia , Ligamento Periodontal/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo
15.
Biochim Biophys Acta ; 1853(3): 561-72, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25523141

RESUMO

Oxysterols, oxidized derivatives of cholesterol, are biologically active molecules. Specific oxysterols have potent osteogenic properties that act on osteoprogenitor cells. However, the molecular mechanisms underlying these osteoinductive effects on embryonic stem cells (ESCs) are unknown. This study investigated the effect of an oxysterol combination of 22(S)-hydroxycholesterol and 20(S)-hydroxycholesterol (SS) on osteogenic differentiation of ESCs and the alterations to mitochondrial activity during differentiation. Osteogenic differentiation was assessed by alkaline phosphatase (ALP) activity, matrix mineralization, mRNA expression of osteogenic factors, runt-related transcription factor 2, osterix, and osteocalcin, and protein levels of collagen type IA (COLIA) and osteopontin (OPN). Treatment of cells with SS increased osteoinductive activity compared to the control group. Intracellular reactive oxygen species production, intracellular ATP content, mitochondrial membrane potential, mitochondrial mass, mitochondrial DNA copy number, and mRNA expression of peroxisome proliferator-activated receptor-γ coactivators 1α and ß, transcription factors involved in mitochondrial biogenesis, were significantly increased during osteogenesis, indicating upregulation of mitochondrial activity. Oxysterol combinations also increased protein levels of mitochondrial respiratory complexes I-V. We also found that SS treatment increased hedgehog signaling target genes, Smo and Gli1 expression. Inhibition of Hh signaling by cyclopamine suppressed mitochondrial biogenesis and ESC osteogenesis. Subsequently, oxysterol-induced Wnt/ß-catenin pathways were inhibited by repression of Hh signaling and mitochondrial biogenesis. Transfection of ß-catenin specific siRNA decreased the protein levels of COLIA and OPN, as well as ALP activity. Collectively, these data suggest that lipid-based oxysterols enhance differentiation of ESCs toward the osteogenic lineage by regulating mitochondrial activity, canonical Hh/Gli, and Wnt/ß-catenin signaling.


Assuntos
Células-Tronco Embrionárias/efeitos dos fármacos , Hidroxicolesteróis/farmacologia , Mitocôndrias/fisiologia , Osteoblastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Células Cultivadas , Transporte de Elétrons/efeitos dos fármacos , Transporte de Elétrons/genética , Células-Tronco Embrionárias/fisiologia , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Proteínas Hedgehog/genética , Proteínas Hedgehog/metabolismo , Camundongos , Osteoblastos/fisiologia , Osteogênese/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética
16.
Phys Chem Chem Phys ; 17(5): 2996-9, 2015 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-25557615

RESUMO

The last decade has seen artificial blood vessels composed of natural polymer nanofibers grafted into human bodies to facilitate the recovery of damaged blood vessels. However, electrospun nanofibers (ENs) of biocompatible materials such as chitosan (CTS) suffer from poor mechanical properties. This study describes the design and fabrication of artificial blood vessels composed of a blend of CTS and PCL ENs and coated with PCL strands using rapid prototyping technology. The resulting tubular vessels exhibited excellent mechanical properties and showed that this process may be useful for vascular reconstruction.


Assuntos
Órgãos Artificiais , Impressão Tridimensional , Materiais Biocompatíveis/química , Vasos Sanguíneos/anatomia & histologia , Vasos Sanguíneos/fisiologia , Quitosana/química , Humanos , Nanofibras/química , Poliésteres/química , Engenharia Tecidual , Alicerces Teciduais
17.
Cell Biochem Funct ; 32(7): 605-11, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25187163

RESUMO

Human periodontal ligament-derived stem cells (PDLSCs) demonstrate self-renewal capacity and multilineage differentiation potential. In this study, we investigated the transdifferentiation potential of human PDLSCs into pancreatic islet cells. To form three-dimensional (3D) clusters, PDLSCs were cultured in Matrigel with media containing differentiation-inducing agents. We found that after 6 days in culture, PDLSCs underwent morphological changes resembling pancreatic islet-like cell clusters (ICCs). The morphological characteristics of PDLSC-derived ICCs were further assessed using scanning electron microscopy analysis. Using reverse transcription-polymerase chain reaction analysis, we found that pluripotency genes were downregulated, whereas early endoderm and pancreatic differentiation genes were upregulated, in PDLSC-derived ICCs compared with undifferentiated PDLSCs. Furthermore, we found that PDLSC-derived ICCs were capable of secreting insulin in response to high concentrations of glucose, validating their functional differentiation into islet cells. Finally, we also performed dithizone staining, as well as immunofluorescence assays and fluorescence-activated cell sorting analysis for pancreatic differentiation markers, to confirm the differentiation status of PDLSC-derived ICCs. These results demonstrate that PDLSCs can transdifferentiate into functional pancreatic islet-like cells and provide a novel, alternative cell population for pancreatic repair.


Assuntos
Transdiferenciação Celular , Ilhotas Pancreáticas/citologia , Ligamento Periodontal/citologia , Células-Tronco/citologia , Linhagem da Célula , Células Cultivadas , Endoderma/citologia , Endoderma/metabolismo , Humanos , Ilhotas Pancreáticas/metabolismo , Dente Molar/citologia , Células-Tronco/metabolismo
18.
Clin Oral Implants Res ; 25(7): 831-7, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23560589

RESUMO

PURPOSE: We evaluated the activities of both osteoblastic and osteoclastic differentiation on sandblasted/acid etched (SLA), hydrophilic SLA surfaces (modSLA) and pretreatment titanium (PT). MATERIAL AND METHODS: The osteoblastic differentiation was evaluated by alkaline phosphatase analysis and Alizarin Red S staining, and the expression of bone-related proteins, alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), osteopontin (OPN), and osteocalcin (OCN), was investigated by reverse transcriptase-polymerase chain reaction (RT-PCR). Primary mice monocytes were expanded and differentiated in the presence of macrophage-colony stimulating factor (M-CSF), and osteoclastic differentiation was evaluated by actin ring formation assay and tartrate-resistant acid phosphatase (TRAP) activity assay. Real-time PCR tests were performed to investigate the expression of gene mRNA expression levels in osteoclast cells. RESULT: Differentiation of osteoblasts in the Alizarin Red S test staining and ALP assay was significantly increased in the modSLA surface. The preceding results were supported by the result of RT-PCR for the expression of Runx2, OPN, and OCN. As for osteoclastic activity, differentiated osteoclasts rarely existed on the SLA and modSLA surface with actin ring. The results of real-time PCR and TRAP activity supported the preceding results. CONCLUSION: It may be concluded that the modSLA surface promotes osteogenic effect and prevents osteoclastic differentiation. Promotion of osteoblastic proliferation after a short-term cell culture might be responsible for stimulated bone regeneration implying that early loading may be possible. Also, the anti-osteoclastic effect of the modSLA surface may contribute to maintenance of the marginal bone level of dental implants, implying long-term stability would be provided by this surface technology. The modSLA surface may not only make early loading possible but possibly reduce marginal bone loss during the maintenance phase.


Assuntos
Corrosão Dentária/métodos , Osteoblastos/citologia , Osteoclastos/citologia , Condicionamento Ácido do Dente , Fosfatase Ácida/metabolismo , Fosfatase Alcalina/metabolismo , Animais , Diferenciação Celular , Células Cultivadas , Interações Hidrofóbicas e Hidrofílicas , Isoenzimas/metabolismo , Fator Estimulador de Colônias de Macrófagos/farmacologia , Camundongos , Microscopia Confocal , Microscopia Eletrônica de Varredura , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Coloração e Rotulagem , Propriedades de Superfície , Fosfatase Ácida Resistente a Tartarato , Titânio
19.
J Nanosci Nanotechnol ; 14(10): 7488-94, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25942814

RESUMO

Electrospun chitosan (CTS) nanofibers have been well known for use as a wound dressing in the biomedical field. Nevertheless, fatal bacterial infections are still a serious problem when CTS nanofibers are used for wound treatment. In this study, we designed a novel wound dressing based on blending the chitosan with polyurethane (CTS/PU) containing silver sulfadiazine (AgSD) in order to enhance both antibacterial activity and mechanical strength. This fiber sheet was produced using the electrospinning (ELSP) technique. The CTS/PU containing AgSD fiber sheet was characterized by energy-dispersive X-ray spectroscopy (EDX). The physicochemical properties of the CTS/PU/AgSD fiber sheets were also characterized by thermogravimetric analysis (TGA) and Fourier transform infrared spectroscopy (FT-IR). The electrospun fibers were morphologically characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). For an in vitro evaluation, the CTS/PU/AgSD fiber sheets were tested for their antibacterial activity against gram-negative Pseudomonas aeruginosa (P. aeruginosa), gram-positive Staphylococcus aureus (S. aureus) and Methicillin-resistant Staphylococcus aureus (MRSA). The results indicate that CTS/PU/AgSD fiber sheets have strong antimicrobial activity as displayed by inhibition of bacterial growth and prevention of infection during the healing process. These results indicate that this material would be good for use as a wound dressing material.


Assuntos
Antibacterianos/química , Antibacterianos/farmacologia , Bandagens/microbiologia , Quitosana/química , Poliuretanos/química , Sulfadiazina de Prata/química , Cicatrização , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/efeitos dos fármacos , Cicatrização/efeitos dos fármacos
20.
J Hazard Mater ; 465: 133036, 2024 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-38000286

RESUMO

A highly efficient system incorporates the real-time visualization of the two toxic molecules (H2S and N2H4) and the recognition of corresponding transforms using a fluorescent sensor. In this paper, a dual-responsive probe (QS-DNP) based on methylquinolinium-salicyaldehyde-2,4-dinitrophenyl was developed that can simultaneously detect H2S and N2H4 at two independent fluorescent channels without signal crosstalk. QS-DNP showed excellent anti-interference, high selectivity, outstanding water solubility, low LOD values (H2S: 51 nM; N2H4: 40 nM), low cytotoxicity, and mitochondrial localization properties. The 2,4-dinitrophenyl site was sensitive to H2S, and the CC bridge was reactive to N2H4, with strong fluorescence at 680 and 488 nm, respectively. The wavelength gap between these two channels is 192 nm; verify that there is no signal crosstalk throughout detection. By this means, the probe was used to simultaneously detect H2S and N2H4 in real soil samples, food samples, and living cells. The endogenous H2S and N2H4 were monitored in HeLa cells and investigated the mitochondria organelle of living cells with a positive charge on QS-DNP. Overall, all results emphasize that the QS-DNP probe is a powerful tool for the simultaneous detection of H2S and N2H4 and presents a potential new sensing approach.


Assuntos
Corantes Fluorescentes , Hidrazinas , Sulfeto de Hidrogênio , Humanos , Células HeLa , Mitocôndrias , Espectrometria de Fluorescência
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