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1.
J Periodontal Res ; 58(2): 403-413, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36653725

RESUMO

BACKGROUND AND OBJECTIVES: Periodontitis is an immunoinflammatory disease characterized by irreversible periodontal attachment loss and bone destruction. Ferroptosis is a kind of immunogenic cell death that depends on the participation of iron ions and is involved in various inflammatory and immune processes. However, information regarding the relationship between ferroptosis and immunomodulation processes in periodontitis is extremely limited. The purpose of this study was to investigate the correlation between ferroptosis and immune responses in periodontitis. METHODS: Gene expression profiles of gingivae were collected from the Gene Expression Omnibus data portal. After detecting differentially expressed ferroptosis-related genes (FRGs), we used univariate logistic regression analysis followed by logistic least absolute shrinkage and selection operator (LASSO) regression to establish a ferroptosis-related classification model in an attempt to accurately distinguish periodontitis gingival tissues from healthy samples. The infiltration level of immunocytes in periodontitis was then assessed through single-sample gene-set enrichment analysis. Subsequently, we screened out immune-related genes by weighted correlation network analysis and protein-protein interaction (PPI) analysis and constructed an immune-related network based on FRGs and immune-related genes. RESULTS: A total of 24 differentially expressed FRGs were detected, and an 8-FRG combined signature constituted the classification model. The established model showed outstanding discriminating ability according to the results of receiver operating characteristic (ROC) curve analysis. In addition, the periodontitis samples had a higher degree of immunocyte infiltration. Activated B cells had the strongest positive correlation while macrophages had a strong negative correlation with certain FRGs, and we found that XBP1, ALOX5 and their interacting genes might be crucial genes in the immune-related network. CONCLUSIONS: The FRG-based classification model had a satisfactory determination ability, which could bring new insights into the pathogenesis of periodontitis. Those genes in the immune-related network, especially hub genes along with XBP1 and ALOX5, would have the potential to serve as promising targets of immunomodulatory treatments for periodontitis.


Assuntos
Ferroptose , Linfócitos B , Gengiva , Nível de Saúde , Imunomodulação
2.
Mikrochim Acta ; 187(7): 377, 2020 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-32519072

RESUMO

Lateral flow immunostrips were newly designed and a sensitive and rapid fluorometric method for the determination of 8-hydroxy-2'-deoxyguanosine (8-OHdG) as a model target of small biomarker molecules was developed. The upconversion nanoparticles (UCNPs, NaYF4:Yb/Er core, and polyacrylic acid (PAA)-modified shell, size ~ 39 nm, excitation wavelength = 980 nm; emission wavelength = 540 nm) were employed as fluorescence signal material. The 8-OHdG antibody (Ab) was taken as the recognition probe while UCNP-labeled Ab was taken as the signal probe. Bovine serum albumin (BSA) was designed as carrier protein for 8-OHdG to form 8-OHdG-BSA conjugate as the capture probe. The lateral flow immunostrips were prepared by laminating a sample pad (glass fiber membrane), a test pad (nitrocellulose membrane), and adsorption pad (filter paper) on PVP backing. The capture probe was immobilized on the test zone while an IgG antibody taken as the control probe was immobilized on the control zone. When the signal probe and the sample were in sequence loaded on the sample pad, 8-OHdG analyte bound with the signal probe, and then the excess of the signal probe move along the strip and is collected by the capture probe on the test zone while the remnant signal probe is collected by the control probe on the control zone. The signal probe and capture probe were synthesized and characterized. The fluorescence intensity on the test zone was inversely proportional to the concentration of 8-OHdG for the quantitative determination while the fluorescence emission on the control zone was observed to validate the assay. The developed method showed a wide linear range from 0.10 to 10 nM, a quite low detection limit of 0.05 nM, small sample volume requirement (100 µL), short assay time (15 min), and good method reproducibility (RSD = 4.4%, nine immunostrips). Graphical abstract Schematic illustration of the configuration and measurement principle of lateral flow fluorescence immunostrip for 8-OHdG: (a) configuration; (b) preparation: load of capture probe (BSA-8-OHdG, 2 µL) on test zone; load of control probe (IgG Ab, 2 µL) on control zone; load of signal probe (UCNP-Ab, 16 µL) on sample pad; (c) measurement: load of sample (8-OHdG, 100 µL) on sample pad, collection, and measurement.


Assuntos
8-Hidroxi-2'-Desoxiguanosina/urina , Imunoensaio/métodos , Nanopartículas/química , 8-Hidroxi-2'-Desoxiguanosina/imunologia , Resinas Acrílicas/química , Anticorpos Imobilizados/imunologia , Érbio/química , Érbio/efeitos da radiação , Fluoretos/química , Fluoretos/efeitos da radiação , Humanos , Imunoensaio/instrumentação , Raios Infravermelhos , Limite de Detecção , Nanopartículas/efeitos da radiação , Testes Imediatos , Reprodutibilidade dos Testes , Itérbio/química , Itérbio/efeitos da radiação , Ítrio/química , Ítrio/efeitos da radiação
3.
J Mater Sci Mater Med ; 23(11): 2717-26, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22903598

RESUMO

The aim of the present work was to design a bio-interactive implant surface by coating recombinant human dentin matrix protein 1 (hDMP1) onto titanium and to investigate the biological function of this material. Firstly, the plasmid containing the hDMP1 cDNA was constructed and hDMP1 was expressed, purified and characterized. Then, hDMP1 was coated onto the surface of Ti substrates via a biochemical technique and the procedure was divided into three steps: in the beginning, titanium was treated by regular polishing and denoted as Cp-Ti; then, Cp-Ti received alkaline and water treatment and was nominated as AW-Ti; finally, AW-Ti was coated with hDMP1 and referred to as hDMP1-Ti. The inserts of hDMP1 genes were detected by enzyme digestion as well as gel electrophoresis, and the complete nucleotide sequence of hDMP1 was tested. The purified recombinant hDMP1 was electrophoresed on a 10 % SDS-PAGE gel. Cp-Ti, AW-Ti and hDMP1-Ti were characterized by X-ray photoelectron spectroscope and water contact angles tests. The biological activity of MG63 cells cultured in the three groups was investigated by the cell attachment, proliferation and alkaline phosphatase activity assays. The results show that hDMP1 was successfully constructed and coated onto the titanium surface, and hDMP1-Ti had higher hydrophilicity than Cp-Ti. Compared with Cp-Ti and AW-Ti, hDMP1-Ti showed better in vitro bioactivity.


Assuntos
Fosfatase Alcalina/metabolismo , Adesão Celular , Proliferação de Células , Proteínas da Matriz Extracelular/química , Fosfoproteínas/química , Sequência de Bases , Linhagem Celular , Primers do DNA , DNA Complementar , Eletroforese em Gel de Poliacrilamida , Proteínas da Matriz Extracelular/genética , Humanos , Dados de Sequência Molecular , Fosfoproteínas/genética , Espectroscopia Fotoeletrônica , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Propriedades de Superfície
4.
In Vitro Cell Dev Biol Anim ; 57(6): 620-630, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34212339

RESUMO

Human dental follicle cells (HDFCs) are an ideal cell source of stem cells for dental tissue repair and regeneration and they have great potential for regenerative medicine applications. However, the conventional monolayer culture usually reduces cell proliferation and differentiation potential due to the continuous passage during in vitro expansion. In this study, primary HDFC spheroids were generated on 1% agarose, and the HDFCs spontaneously formed cell spheroids in the agarose-coated dishes. Compared with monolayer culture, the spheroid-derived HDFCs exhibited increased proliferative ability for later passage HDFCs as analysed by Cell Counting Kit-8 (CCK-8). The transcription-quantitative polymerase chain reaction (qRT-PCR), western blot and immunofluorescence assay showed that the expression of stemness marker genes Sox2, Oct4 and Nanog was increased significantly in the HDFC spheroids. Furthermore, we found that the odontogenic differentiation capability of HDFCs was significantly improved by spheroid culture in the agarose-coated dishes. On the other hand, the osteogenic differentiation capability was weakened compared with monolayer culture. Our results suggest that spheroid formation of HDFCs in agarose-coated dishes partially restores the proliferative ability of HDFCs at later passages, enhances their stemness and improves odontogenic differentiation capability in vitro. Therefore, spheroid formation of HDFCs has great therapeutic potential for stem cell clinical therapy.


Assuntos
Técnicas de Cultura de Células , Saco Dentário/crescimento & desenvolvimento , Odontogênese/efeitos dos fármacos , Esferoides Celulares/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Proliferação de Células/efeitos dos fármacos , Saco Dentário/citologia , Saco Dentário/metabolismo , Humanos , Odontogênese/genética , Sefarose/farmacologia , Esferoides Celulares/citologia , Células-Tronco/efeitos dos fármacos
5.
Colloids Surf B Biointerfaces ; 93: 226-34, 2012 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-22305638

RESUMO

In this study, the multiwall carbon nanotubes (MWNTs)/polycaprolactone composite scaffolds were fabricated by the solution evaporation technique. The morphology, phase composition and the mechanical properties of the composite scaffolds were characterized and the cellular bioactivity of the scaffolds was assessed by using rat bone-marrow-derived stroma cells (BMSCs). The attachment, proliferation and differentiation of the BMSCs on the composite scaffolds were analyzed by scanning electron microscopy (SEM), 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) nuclear staining and fluorescein diacetate (FDA) and propidium iodide (PI) live/dead staining, methylthiazol tetrazolium (MTT) assay and alkaline phosphatase (ALP) activity assay, respectively. Results showed that mechanical properties of the composite scaffolds were improved with the addition of MWNTs (0.25-2 wt%). BMSCs on the composite scaffolds differentiated down the osteogenic lineage and expressed high levels of bone marker ALP. The scaffolds with low concentration (0.5 wt%) of MWNTs can enhance the proliferation and differentiation of the BMSCs more than that with higher concentration of MWNTs. It is concluded that MWNTs/PCL composite scaffolds have the potential for bone tissue engineering and the relatively low concentration of MWNTs (0.5 wt%) is preferred.


Assuntos
Células da Medula Óssea/efeitos dos fármacos , Células-Tronco Mesenquimais/efeitos dos fármacos , Nanocompostos/química , Poliésteres/química , Engenharia Tecidual , Fosfatase Alcalina/metabolismo , Animais , Materiais Biocompatíveis , Células da Medula Óssea/citologia , Células da Medula Óssea/fisiologia , Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Indóis , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/fisiologia , Microscopia Eletrônica de Varredura , Nanocompostos/ultraestrutura , Nanotubos de Carbono/química , Nanotubos de Carbono/ultraestrutura , Poliésteres/farmacologia , Ratos , Alicerces Teciduais
6.
Hua Xi Kou Qiang Yi Xue Za Zhi ; 30(2): 222-3, 2012 Apr.
Artigo em Zh | MEDLINE | ID: mdl-22594249

RESUMO

It is to address torquing an individual tooth using a gate spring. The gate spring is made of a rectangular stainless steal wire, in the shape of a gate, which is incorporated to the archwire by spot welding. Torque is generated by the combined effects of the gate spring and the archwire. After 2-3 months, the gate spring can obviously torque individual tooth.


Assuntos
Desenho de Aparelho Ortodôntico , Fios Ortodônticos , Braquetes Ortodônticos , Dente , Técnicas de Movimentação Dentária , Torque
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