RESUMEN
BACKGROUND: Staphylococcus arlettae is a rarely reported coagulase-negative staphylococcus (CoNS) isolated from infected humans and livestock. Observing phage-bacteria interaction could improve the understanding of bacterial pathogenetic mechanisms, providing foundational evidence for phage therapy or phage detection. Herein, we aimed to characterise and annotate a novel bacteriophage, vB_SarS_BM31 (BM31), specific to S. arlettae. This bacteriophage was isolated from a milk sample associated with bovine mastitis and collected in the Sichuan Province, China. RESULTS: The BM31 genome comprised a linear double-stranded DNA of 42,271 base pair in length with a G + C content of 34.59%. A total of 65 open reading frames (ORFs) were assembled from phage DNA, of which 29 were functionally annotated. These functional genes were divided into four modules: the structural, DNA packing and replication, lysis, and lysogeny modules. Holin (ORF25), lysin (ORF26), and integrase (ORF28) were located closely in the entire BM31 genome and were important for lyse or lysogeny cycle of BM31. The phage was identified as a temperate phage according to whole genome analysis and life cycle assay, with basic biological characteristics such as small burst size, short latency period, and narrow host range, consistent with the characteristics of the family Siphoviridae, subcluster B14 of the Staphylococcus bacteriophage. CONCLUSIONS: The present isolation and characterisation of BM31 contributes to the Staphylococcus bacteriophage database and provides a theoretical foundation for its potential applications. To the best of our knowledge, BM31 is the only shared and completely reported phage against S. arlettae in the entire public database.
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ADN Viral , Genoma Viral , Animales , Bovinos , Femenino , Humanos , Análisis de Secuencia de ADN , ADN Viral/genética , ADN Viral/química , Staphylococcus/genética , Fagos de Staphylococcus/genéticaRESUMEN
OBJECTIVE: To verify the role of YAP/WNT5A/FZD4 axis in stretch-induced osteogenic differentiation of hPDLCs. BACKGROUND: During orthodontic tooth movement, differentiation of human periodontal ligament cells (hPDLCs) at the tension side of the periodontal ligament mediates new bone formation. WNT5A promotes osteogenesis and its regulator Yes-associated protein (YAP) is responsive to mechanical stimulation in hPDLCs. However, the mechanisms of YAP and WNT5A in alveolar bone remodeling remain unclear. METHODS: Cyclic stretch was applied to hPDLCs to mimic the orthodontic stretching force. Osteogenic differentiation was determined by alkaline phosphatase (ALP) activity, Alizarin Red staining, qRT-PCR and western blotting. To detect activation of YAP and expression of WNT5A and its receptor Frizzled-4 (FZD4), western blotting, immunofluorescence, qRT-PCR and ELISA were performed. Verteporfin, Lats-IN-1, small interfering RNAs and recombinant protein were used to explore the relationship of YAP, WNT5A and FZD4, and the effect of their relationship on stretch-induced osteogenesis of hPDLCs. RESULTS: WNT5A, FZD4 and nuclear localization of YAP were upregulated by cyclic stretch. YAP positively regulated WNT5A and FZD4 expression and osteogenic differentiation of hPDLCs under cyclic stretch by YAP inhibition or activation assay. Knockdown of WNT5A and FZD4 attenuated YAP-induced and stretch-induced osteogenic differentiation. Recombinant WNT5A rescued the suppressed osteogenic differentiation by YAP inhibitor in hPDLCs, whereas knockdown of FZD4 weakened the effect of WNT5A and amplified the suppression. CONCLUSIONS: WNT5A/FZD4 could be positively regulated by YAP and the YAP/WNT5A/FZD4 axis mediated osteogenic differentiation of hPDLCs under cyclic stretch. This study provided further insight into the biological mechanism of orthodontic tooth movement.
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Osteogénesis , Ligamento Periodontal , Humanos , Células Cultivadas , Diferenciación Celular , Proteínas/metabolismo , Proteína Wnt-5a/metabolismo , Receptores Frizzled/metabolismoRESUMEN
Osteogenic differentiation and bone formation are tightly regulated by several factors, including microRNAs (miRNAs). However, miRNA expression patterns and function during mechanical loading-induced osteogenic differentiation of human periodontal ligament cells (PDLCs) remain unclear. Here, we investigated the differential expression of miRNA-195-5p in the periodontal tissues of mice under orthodontic mechanical loading and in primary human PDLCs exposed to a simulated tension strain. The miR-195-5p was observed to be down-regulated and negatively correlated with osteogenic differentiation. Overexpression of miR-195-5p significantly inhibited PDLC differentiation under cyclic tension strain (CTS), whereas the functional inhibition of miR-195-5p yielded an opposite effect. Further experiments confirmed that WNT family member 3A (WNT3A), fibroblast growth factor 2 (FGF2), and bone morphogenetic protein receptor-1A (BMPR1A), proteins important for osteogenic activity and stability, were direct targets of miR-195-5p. Mechanical loading increased the WNT3A, FGF2, and BMPR1A protein levels, while miR-195-5p inhibited WNT3A, FGF2, and BMPR1A protein expression. WNT, FGF, and BMP signaling were involved in osteogenic differentiation of PDLCs under CTS. Further study confirmed that reintroduction of WNT3A and BMPR1A can rescue the inhibition of miR-195-5p on osteogenic differentiation of PDLCs. Our findings are the first to demonstrate that miR-195-5p is a mechanosensitive gene that plays an important role in mechanical loading-induced osteogenic differentiation and bone formation.
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Diferenciación Celular , Mecanotransducción Celular , Osteogénesis , Ligamento Periodontal/metabolismo , Regiones no Traducidas 3' , Adolescente , Animales , Sitios de Unión , Receptores de Proteínas Morfogenéticas Óseas de Tipo 1/genética , Receptores de Proteínas Morfogenéticas Óseas de Tipo 1/metabolismo , Células Cultivadas , Factor 2 de Crecimiento de Fibroblastos/genética , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Regulación de la Expresión Génica , Humanos , Ratones , Ratones Endogámicos C57BL , MicroARNs/genética , MicroARNs/metabolismo , Modelos Animales , Aparatos Ortodóncicos , Ligamento Periodontal/patología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Transducción de Señal , Estrés Mecánico , Factores de Tiempo , Transfección , Proteína Wnt3A/genética , Proteína Wnt3A/metabolismo , Adulto JovenRESUMEN
Sixty-nine experienced Chinese orthodontists evaluated 108 Chinese patients' facial attractiveness from set of photographs (frontal, lateral, and frontal smiling photos) taken at the end of orthodontic treatment. These 108 patients, which contained an equal number of patients with Class I, II, and III malocclusion, were randomly selected from 6 orthodontic treatment centers throughout China. Spearman rank-order correlation coefficients (rs) analyses were performed to examine agreement in ranking between all judge pairs. Pearson correlation and multivariate regression were performed to examine the correlation between cephalometric measures and end-of-treatment Photo Attractiveness Rank.96.68% judge pairs showed moderate correlated (+0.4â≤ârsâ<â+0.7) subjective rankings. Cephalometric measures significantly correlated with end-of-treatment Photo Attractiveness Rank included interincisal angle (râ=â0.330, Pâ<â0.05), L1/MP° (râ=â0.386, Pâ<â0.05), L1-NBmm (râ=â0.451, Pâ<â0.01), L1/NB° (râ=â0.374, Pâ<â0.05), and profile angle (râ=â0.353, Pâ<â0.05) in Class I patients with an explained variance of 32.8%, and ANB angle (râ=â0.432, Pâ<â0.01), angle of convexity (râ=â0.448, Pâ<â0.01), profile angle (râ=â0.488, Pâ<â0.01), Li to E-line (râ=â0.374, Pâ<â0.05), Li to B-line (râ=â0.543, Pâ<â0.01), and Z angle (râ=â0.543, Pâ<â0.01) in Class II patient with an explained variance of 43.3%.There was less association than expected between objective measurements on the lateral cephalograms and clinicians' rankings of facial attractiveness on clinical photography in Chinese patients. Straight-stand lower incisor was desired for facial attractiveness of Class I malocclusion; and sagittal relationship and lip prominence influence the esthetics of Class II malocclusion in Chinese population.
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Cefalometría/métodos , Estética Dental , Cara/anatomía & histología , Maloclusión/terapia , Adolescente , Adulto , Niño , China , Femenino , Humanos , Masculino , Maloclusión/diagnóstico , Sonrisa , Adulto JovenRESUMEN
The bone formation (osteogenesis) of human periodontal ligament cells (hPDLCs) under tension stress is essential for alveolar bone remodeling during orthodontic tooth movement (OTM). Deubiquitinating enzymes (DUBs) remove ubiquitin from target proteins, affecting their function and mediating cell survival and differentiation. However, whether and how DUBs regulate hPDLC function under tension force is poorly understood. In this study, we first investigated the expression of DUBs in hPDLCs under cyclic tension stimulation (CTS). Up-regulation of USP12 was observed in hPDLCs and at the tension side of molar teeth in OTM C57BL6 mice models. Knockdown (KD) of USP12 led to enhanced osteogenesis of hPDLCs under CTS. RNA-seq analysis suggested that the unfolded protein response (UPR) was the prevailing biological process in hPDLCs with USP12 KD, indicating that USP12 depletion triggered endoplasmic reticulum (ER) stress. The three major UPR-related signaling branches, namely PERK/eIF2α/ATF4, IRE1α/XBP1s, and ATF6 axis, were activated in hPDLCs with USP12 KD. By utilizing specific inhibitors, we proved that the PERK/eIF2α/ATF4 axis predominantly mediated the enhanced osteogenesis in hPDLCs with USP12 KD under CTS. In summary, our study demonstrates that USP12 serves as a key regulator for CTS-induced osteogenesis in hPDLCs, suggesting that USP12 upregulation serves as an adaptive mechanism for hPDLCs to alleviate ER stress during OTM.
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Osteogénesis , Ligamento Periodontal , Humanos , Animales , Ratones , Osteogénesis/fisiología , Endorribonucleasas/metabolismo , Ratones Endogámicos C57BL , Proteínas Serina-Treonina Quinasas/metabolismo , Diferenciación Celular/fisiología , Células Cultivadas , Ubiquitina Tiolesterasa/metabolismoRESUMEN
Background and Objectives: To investigate the role of exosomes from periodontal ligament cells (PDLCs) in bone marrow mesenchymal stem cell (BMSC) migration. Methods and Results: Human PDLCs were applied cyclic tension stretching. Exosomes were extracted from cultured PDLCs by ultracentrifugation, then characterized for their size, morphology and protein markers by NTA, TEM and western blotting. The process that PKH26-labeled exosomes taken up by BMSCs was assessed by confocal microscope. BMSC migration was examined by Transwell assay. Exosomes derived from PDLCs were identified. Cyclic tension stretch application on PDLCs can enhance the migration ability of BMSCs through exosomes. The exosomal miRNA expression profiles of unstretched and stretched PDLCs were tested by miRNA microarray. Four miRNAs (miR-4633-5p, miR-30c-5p, miR-371a-3p and let-7b-3p) were upregulated and six (miR-4689, miR-8485, miR-4655-3p, miR-4672, miR-3180-5p and miR-4476) were downregulated in the exosomes after stretching. Sixteen hub proteins were found in the miRNA-mRNA network. Gene Ontology and KEGG pathway analyses demonstrated that the target genes of differentially expressed exosomal miRNAs closely related to the PI3K pathway and vesicle transmission. Conclusions: The exosomes derived from cyclic tension-stretched PDLCs can promote the migration of BMSCs. Alternation of microRNA profiles provides a basis for further research on the regulatory function of the exosomal miRNAs of PDLCs during orthodontic tooth movement.
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A boy, aged 12 years 3 months, sought treatment for a complete unilateral Brodie bite. His maxillary dental arch was asymmetric, and his dentition was 10 mm wider than normal values for his age. The transverse discrepancy was his chief complaint because it caused a chewing dysfunction. We used a special slow maxillary contraction appliance, which contains a screw, connectors, and retainers, to contract the maxillary transverse asymmetric dental arch. This was followed by preadjusted fixed appliances to level the teeth and adjust the occlusion. A symmetric and functional Class I occlusion was achieved.
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Arco Dental/patología , Diseño de Aparato Ortodóncico , Niño , Humanos , Masculino , Maloclusión/terapiaRESUMEN
With the improper use of antibiotics, an increasing number of multidrug-resistant bacteria have been reported worldwide, posing challenges for disease treatment. Klebsiella pneumoniae is an important zoonotic pathogen that colonises the respiratory tract. Endolysin therapy has emerged with the development of phages. In this study, a lytic phage vB_KpnS_MK54 was isolated from the drinking water of a forest musk deer (FMD) farm in Sichuan Province. It was the first reported phage obtained from FMD. The primary biological characteristics were determined, and whole-genome sequencing analysis was performed. The phage which belongs to the family Siphoviridae is highly specific for lytic host bacteria and is moderately adaptable to different environments. Whole-genome sequencing results showed that the phage genome size was 46,218 bp. There were 80 coding DNA sequences (CDSs) in total, 32 of which had known functions. The last CDS is the phage endolysin LysG24. A new peptide-modified endolysin (LysCA) was constituted by connecting the cecropin A peptide residues with LysG24 to investigate the antibacterial activities of both LysG24 and LysCA. The results showed that the lytic profile of LysG24 and LysCA was wider than that of phage MK54. For in vitro tests, both endolysins destroyed 99% of the host bacteria within 6 h. The lysing ability and environmental adaptability of LysCA were significantly stronger than those of LysG24. For in vivo tests, LysG24 and LysCA exhibited therapeutic effects in a mouse model of pneumonia wherewith the mice were infected with K. pneumoniae (LPKP), wherein both LysG24 and LysCA can effectively reduce the pulmonary inflammatory response. The LPKP bacterial load in the treatment group was significantly lower than that in the bacterial group, among which LysCA displayed a more obvious therapeutic effect. Furthermore, the safety test showed that the endolysins had no toxic effects on mice. In general, both LysG24 and LysCA showed excellent antibacterial activity in vivo and in vitro, with high safety and strong adaptability to the environment, manifesting their latent potential as new antimicrobial agents.
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Bovine mastitis is a common disease occurring in dairy farms and can be caused by more than 150 species of pathogenic bacteria. One of the most common causative organisms is Streptococcus agalactiae, which is also potentially harmful to humans and aquatic animals. At present, research on S. agalactiae in China is mostly concentrated in the northern region, with limited research in the southeastern and southwestern regions. In this study, a total of 313 clinical mastitis samples from large-scale dairy farms in five regions of Sichuan were collected for isolation of S. agalactiae. The epidemiological distribution of S. agalactiae was inferred by serotyping isolates with multiplex polymerase chain reaction. Susceptibility testing and drug resistance genes were detected to guide the clinical use of antibiotics. Virulence genes were also detected to deduce the pathogenicity of S. agalactiae in Sichuan Province. One hundred and five strains of S. agalactiae (33.6%) were isolated according to phenotypic features, biochemical characteristics, and 16S rRNA sequencing. Serotype multiplex polymerase chain reaction analysis showed that all isolates were of type Ia. The isolates were up to 100% sensitive to aminoglycosides (kanamycin, gentamicin, neomycin, and tobramycin), and the resistance rate to ß-lactams (penicillin, amoxicillin, ceftazidime, and piperacillin) was up to 98.1%. The TEM gene (ß-lactam-resistant) was detected in all isolates, which was in accordance with a drug-resistant phenotype. Analysis of virulence genes showed that all isolates harbored the cfb, cylE, fbsA, fbsB, hylB, and α-enolase genes and none harbored bac or lmb. These data could aid in the prevention and control of mastitis and improve our understanding of epidemiological trends in dairy cows infected with S. agalactiae in Sichuan Province.
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Mastitis Bovina , Infecciones Estreptocócicas , Animales , Antibacterianos/farmacología , Bovinos , Farmacorresistencia Microbiana , Femenino , Humanos , Mastitis Bovina/epidemiología , Mastitis Bovina/microbiología , Tipificación Molecular , Prevalencia , ARN Ribosómico 16S , Infecciones Estreptocócicas/epidemiología , Infecciones Estreptocócicas/microbiología , Infecciones Estreptocócicas/veterinaria , Streptococcus agalactiae , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
Klebsiella pneumoniae is an important pathogen that can infect both humans and cattle. The widespread K. pneumoniae and its high drug resistance make it difficult to treat Klebsiella infections/diseases. In this study, a lytic K. pneumoniae bacteriophage vB_Kpn_B01 was isolated from a dairy farm trough in Sichuan Province, and its biological properties were studied, and the entire genome of vB_Kpn_B01 was sequenced. The therapeutic effects of the phage on disease-causing mice were preliminarily tested. Phages found in this study are double-stranded DNA bacterial viruses belonging to the family Siphoviridae, Sugarlandvirus. The results suggest that vB_Kpn_B01 has strong specificity and low adaptability to different adverse conditions. Meanwhile, the predicted gene products of phage vB_Kpn_B01 comprised 149 coding sequences (CDS) and 25 tRNAs, of which 34 CDS had known functions. Of course, vB_Kpn_B01 did not contain any known antibiotic-resistant or virulent genes. The pathological sections of the liver and lungs of mice showed that the inflammatory scores of the treatment group were lower than in the bacterial group. Phage vB_Kpn_B01 alleviated the inflammatory response in the organs of the infected mice, and the organ tissue bacterial load of the treatment group was significantly lower than that of the bacterial group. Therefore, vB_Kpn_B01 can inhibit the proliferation of K. pneumoniae 18 in vivo and can alleviate the inflammation of target organs caused by infectious bacteria, which preliminarily indicates that vB_Kpn_B01 has a certain therapeutic effect on laboratory-infected mice.
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In this study, porous chitosan/collagen scaffolds were prepared through a freeze-drying process, and loaded with the plasmid vector encoding human bone morphogenetic protein-7 (BMP-7) gene. To investigate the feasibility and efficacy of this gene-activated scaffold on dental tissue engineering, human dental pulp stem cells (DPSCs) were seeded in this scaffold for in vitro and in vivo study. In vitro results indicated that cells can be transfected successfully by loaded plasmid and secrete BMP-7 until day 24. Evaluation of DNA content, ALP activity, calcium content, SEM, and real-time PCR revealed that cells on gene-activated scaffold showed better proliferation properties and odontoblastic differentiation behaviors than cells on pure scaffolds. Then, these cell-scaffold complexes were implanted subcutaneously and retrieved after 4 weeks for histology evaluation. In vivo results that gene-activated scaffold group could still trace the existence of tranfected cells at week 4 and showed the upregulated expression of DSPP compared to pure scaffold groups. On the basis of our results, chitosan/collagen-loaded BMP-7 DNA appears to be an effective substrate candidate for gene delivery and indeed enhanced DPSCs differentiation toward an odontoblast-like phenotype in vitro and in vivo. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A:, 2012.
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Proteína Morfogenética Ósea 7 , Diferenciación Celular/efectos de los fármacos , Colágeno/química , ADN , Pulpa Dental/metabolismo , Odontoblastos/metabolismo , Células Madre/metabolismo , Adolescente , Adulto , Animales , Proteína Morfogenética Ósea 7/química , Proteína Morfogenética Ósea 7/farmacología , ADN/química , ADN/farmacología , Femenino , Xenoinjertos , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Trasplante de Células MadreRESUMEN
During the orthodontic tooth movement, cells in periodontal ligament could differentiate into osteoblasts to synthesize alveolar bone as well as affect the proliferation, migration and differentiation of mesenchymal stem cells, which also contribute to bone remodeling. However, the mechanism is still largely elusive. Here, we evaluated the expression of CREB at the tension site of mouse periodontal ligament under orthodontic mechanical strain and in the cyclic tension strain treated human periodontal ligament cells. Then, through gain and loss of function analysis, we revealed that CREB in PDLCs promotes SDF-1 and FGF2 secretion, which enhance the migration and osteoblastic differentiation of BMSCs. We further discovered that CREB transcriptionally activates FGF2 and SDF-1 expressions by binding to the promoter regions.In conclusion, this study confirms that CREB is an upregulated gene in periodontal ligament under orthodontic tension strain stimulation and plays an important role in regulating BMSCs' physiological activity in orthodontic tension strain-induced bone formation.
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Diferenciación Celular , Movimiento Celular , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Fenómenos Mecánicos , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Técnicas de Movimiento Dental , Fenómenos Biomecánicos , Humanos , Estrés MecánicoRESUMEN
Periodontal remodeling and alveolar bone resorption and formation play essential roles during orthodontic tooth movement (OTM). In the process, human periodontal ligament cells (HPDLCs) sense and respond to orthodontic forces, contributing to the alveolar bone formation. However, the underlying mechanism in this process is not fully elucidated. In the present study, cyclic stress stimulus was applied on HPDLCs to mimic the orthodontic forces during OTM. Our results demonstrated that cyclic stretch promoted the osteogenic differentiation of HPDLCs. Moreover, our data suggested that yes-associated protein (YAP), the Hippo pathway effector, which also involved in mechanical signaling transduction, was activated as we found that the nuclear translocation of YAP was significantly increased in the cyclic stress treated HPDLCs. The mRNA expression of CTGF and CYR61, the target genes of YAP, was also remarkably increased. Furthermore, knockdown of YAP suppressed the cyclic stretch induced osteogenesis in HPDLCs, while overexpression of YAP in HPDLCs enhanced osteogenesis. We also noticed that YAP activities could be suppressed by the ROCK and nonmuscle myosin II inhibitors, Y-27632 and Blebbistatin. The inhibitors also significantly inhibited the cyclic stretch induced osteogenesis in HPDLCs. Finally, in the murine OTM model, our results revealed that YAP was upregulated and nuclearly translocated in the PDLCs at the tension side. In summary, our present study demonstrated that cytoskeleton remodeling induced activation of YAP signaling pathway was crucial for the cyclic stretch-induced osteogenesis of HPDLCs, which might play important roles during OTM.
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Proteínas Adaptadoras Transductoras de Señales/metabolismo , Diferenciación Celular/fisiología , Osteogénesis/fisiología , Ligamento Periodontal/metabolismo , Ligamento Periodontal/fisiología , Fosfoproteínas/metabolismo , Adolescente , Adulto , Amidas/uso terapéutico , Animales , Resorción Ósea/tratamiento farmacológico , Resorción Ósea/metabolismo , Resorción Ósea/fisiopatología , Diferenciación Celular/efectos de los fármacos , Células Cultivadas , Niño , Citoesqueleto/efectos de los fármacos , Citoesqueleto/metabolismo , Citoesqueleto/fisiología , Femenino , Humanos , Masculino , Mecanotransducción Celular/efectos de los fármacos , Mecanotransducción Celular/fisiología , Ratones , Ratones Endogámicos C57BL , Osteogénesis/efectos de los fármacos , Ligamento Periodontal/efectos de los fármacos , Piridinas/uso terapéutico , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Estrés Mecánico , Técnicas de Movimiento Dental/métodos , Factores de Transcripción , Proteínas Señalizadoras YAP , Adulto JovenRESUMEN
Background: Staphylococcus arlettae is a rarely reported coagulase-negative staphylococcus (CoNS) isolated from infected humans and livestock. Observing phage-bacteria interaction could improve the understanding of bacterial pathogenetic mechanisms, providing foundational evidence for phage therapy or phage detection. Herein, we aimed to characterise and annotate a novel bacteriophage, vB_SarS_BM31 (BM31), specific to S. arlettae. This bacteriophage was isolated from a milk sample associated with bovine mastitis and collected in the Sichuan Province, China. Results: The BM31 genome comprised a linear double-stranded DNA of 42,271 base pair in length with a G + C content of 34.59%. A total of 65 open reading frames (ORFs) were assembled from phage DNA, of which 29 were functionally annotated. These functional genes were divided into four modules: the structural, DNA packing and replication, lysis, and lysogeny modules. Holin (ORF25), lysin (ORF26), and integrase (ORF28) were located closely in the entire BM31 genome and were important for lyse or lysogeny cycle of BM31. The phage was identified as a temperate phage according to whole genome analysis and life cycle assay, with basic biological characteristics such as small burst size, short latency period, and narrow host range, consistent with the characteristics of the family Siphoviridae, subcluster B14 of the Staphylococcus bacteriophage. Conclusions: The present isolation and characterisation of BM31 contributes to the Staphylococcus bacteriophage database and provides a theoretical foundation for its potential applications. To the best of our knowledge, BM31 is the only shared and completely reported phage against S. arlettae in the entire public database.(AU)
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Humanos , Lisogenia , 34726 , Integrasas , Mastitis , Staphylococcus , GenomaRESUMEN
Two of the challenges for clinical implementation of nano-therapeutic strategies are optimization of tumor targeting and clearance of the nanoagents in vivo. Herein, a cell-mediated therapy by transporting 2D Bi2Se3 nanosheets within macrophage vehicles is described. The Bi2Se3 nanosheets with excellent near-infrared photothermal performance exhibit high macrophage uptake and negligible cytotoxicity thus facilitating the fabrication of Bi2Se3-laden-macrophages. Compared with bare Bi2Se3, the Bi2Se3-laden-macrophages after intravenous injection show prolonged blood circulation and can overcome the hypoxia-associated drug delivery barrier to target the tumor efficiently and dramatically enhance the efficiency of photothermal cancer therapy. The Bi2Se3-laden-macrophages possess good biocompatibility as demonstrated by the biochemical and histological analyses and furthermore, most of the materials are excreted from the body within 25 days. Our findings reveal a desirable system for highly efficient near-infrared photothermal cancer therapy.
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Nanoestructuras/química , Nanoestructuras/uso terapéutico , Fototerapia/métodos , Animales , Proliferación Celular/efectos de los fármacos , Femenino , Humanos , Hipertermia Inducida/métodos , Rayos Infrarrojos , Células MCF-7 , Neoplasias Mamarias Animales/terapia , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Nanopartículas/química , Células RAW 264.7RESUMEN
Orthodontic tooth movement (OTM) is associated with bone remodeling mediated by orthodontic mechanical loading. Increasing studies reported that Wnt signaling played crucial roles in mechanical stimuli induced bone remodeling. However, little is known about the involvement of Wnt signaling in orthodontic force-induced bone formation during OTM. In virtue of the OTM mice model as we previously reported, where new bone formation was determined by micro-CT and immunoreactivity of osteocalcin and osterix, we explored the activation of Wnt signaling pathway during OTM. Our results proved the nuclei translocation of ß-catenin, suggesting the activation of canonical Wnt signaling pathway in the periodontal ligament cells (PDLCs) near the alveolar bone at the tension site (TS). Moreover, the immunoreactivity of Wnt5a, but not Wnt3a in PDLCs indicated the activation of canonical Wnt pathway might be mediated by Wnt5a, but not Wnt3a as in most cases. The co-location of Wnt5a and ß-catenin that was evidenced by double labeling immunofluorescence staining further supported the hypothesis. In addition, the high expression of FZD4 and LRP5 in PDLCs at TS of periodontium suggested that the activation of Wnt signaling pathway was mediated by these receptors. The negligible expression of ROR2 also indicated that canonical but not non-canonical Wnt signaling pathway was activated by Wnt5a, since previous studies demonstrated that the activation of canonical/non-canonical Wnt signaling pathway was largely dependent on the receptors. In summary, we here reported that Wnt5a mediated activation of canonical Wnt signaling pathway might contribute to the orthodontic force induced bone remodeling.
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Remodelación Ósea , Osteogénesis , Diente/crecimiento & desarrollo , Diente/metabolismo , Vía de Señalización Wnt , Proteína Wnt-5a/genética , Proteína Wnt-5a/metabolismo , Animales , Biomarcadores , Receptores Frizzled/metabolismo , Inmunohistoquímica , Proteína-5 Relacionada con Receptor de Lipoproteína de Baja Densidad/metabolismo , Masculino , Fenómenos Mecánicos , Ratones , Modelos Animales , Receptores Huérfanos Similares al Receptor Tirosina Quinasa/metabolismo , Proteínas Wnt/metabolismo , beta Catenina/metabolismoRESUMEN
A hexagonal onplant of 7.7 mm diameter was placed on the palatal bone of the maxilla in an 1-year five-month-old female patient with a Class III malocclusion and midface deficiency. Elastic traction (400 g per side) was applied from a facemask to the onplant at 30 degrees to the occlusal plane 12 hours per day for 12 months. The maxilla was found to have displaced forward and downward by 2.9 mm. The mandible was rotated downward and backward. There was a 3 degrees increase in mandibular plane angle and an increase in the lower face height. Clinically, there was a significant improvement in midface esthetics, noted by an increase in fullness of the infraorbital region and correction of the skeletal discrepancy between the maxillary and mandibular jaw relationship. Contrary to the reports that use teeth rather than onplants as anchorage, there was no forward movement of the maxillary molars and minimal extrusion of the maxillary molars. These results suggest that onplants can be used as an extremely stable anchorage for maxillary orthopedic facemask treatment.
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Aparatos de Tracción Extraoral , Maloclusión de Angle Clase III/terapia , Técnica de Expansión Palatina/instrumentación , Cefalometría , Niño , Arco Dental/anatomía & histología , Femenino , Humanos , Maloclusión/terapia , Mandíbula/anatomía & histología , Maxilar/anatomía & histología , Diseño de Aparato Ortodóncico , Hueso Paladar/cirugía , Resultado del TratamientoRESUMEN
The aim of this investigation was to compare root resorption in the same individual after application of continuous intrusive and extrusive forces. In nine patients (mean age 15.3 years), the maxillary first premolars were randomly intruded or extruded with a continuous force of 100 cN for eight weeks. Eleven maxillary first premolars from six randomly selected orthodontic patients served as controls. Root resorption was determined using scanning electron microscopy. Quantitative assessment of the percentage of resorbed area of the total root surface was performed on composite micrographs. The severity of root resorption was also assessed by visual scoring of the roots. Root resorption mainly occurred at the apical part of the roots in both experimental groups. A significant difference in root resorption was found between the intruded and the control teeth (P = .006) but not between the extruded and the control teeth. However, the mesial and distal root surfaces showed resorption on 5.78 +/- 3.86% of the root surface of the intruded teeth and 1.28 +/- 1.24% of the root surface of the extruded teeth, and this difference was significant (P = .004). In addition, a large individual variation was found. From this study, it can be concluded that intrusion of teeth causes about four times more root resorption than extrusion. Because the amount of root resorption due to intrusion or extrusion in the same patient is correlated, every clinician should be aware that the extrusion of teeth might also cause root resorption in susceptible patients.
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Resorción Radicular/etiología , Técnicas de Movimiento Dental , Adolescente , Adulto , Diente Premolar/patología , Niño , Femenino , Humanos , Masculino , Microscopía Electrónica de Rastreo , Alambres para Ortodoncia , Planificación de Atención al Paciente , Extracción Seriada , Estrés Mecánico , Ápice del Diente/patología , Técnicas de Movimiento Dental/instrumentación , Técnicas de Movimiento Dental/métodos , Raíz del Diente/patologíaRESUMEN
Tension force-induced bone formation is a complex biological process altered by various factors, for example miRNAs and gene regulatory network. However, we know little about critical gene regulators and their functional consequences on this complex process. The aim of this study was to determine the integrated relation between microRNA and mRNA expression in tension force-induced bone formation in periodontal ligament cells by a system biological approach. We identified 818 mRNAs and 32 miRNAs differentially expressed between cyclic tension force-stimulated human periodontal ligament cells and control cells by microarrays. By using miRNA/mRNA network analysis, protein-protein interactions network analysis, and hub analysis, we found that miR-195-5p, miR-424-5p, miR-1297, miR-3607-5p, miR-145-5p, miR-4328, and miR-224-5p were core microRNAs of tension force-induced bone formation. WDR33, HSPH1, ERBB3, RIF1, IKBKB, CREB1, FGF2, and PAG1 were identified as hubs of the PPI network, suggesting the biological significance in this process. The miRNA expression was further examined in human PDLC and animal samples by using quantitative real-time PCR. Thus, we proposed a model of tension force-induced bone formation which is co-regulated through integration of the miRNA and mRNA. This study illustrated the benefits of system biological approaches in the analysis of tension force-induced bone formation as a complex biological process. We used public information and our experimental data to do comprehensive analysis and revealed the coordination transcriptional control of miRNAs of tension force-induced bone formation.
Asunto(s)
MicroARNs/genética , Ligamento Periodontal/citología , ARN Mensajero/genética , Animales , Femenino , Regulación de la Expresión Génica/fisiología , Humanos , Técnicas In Vitro , Ratones , Ratones Endogámicos C57BL , Osteogénesis/genética , Osteogénesis/fisiología , Ligamento Periodontal/fisiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Estrés Mecánico , Análisis de Matrices Tisulares , Movilidad DentariaRESUMEN
OBJECTIVE: The ideal denture adhesive demonstrates a neutral or slightly basic pH, satisfactory bonding load, and minimal toxicity to the oral mucosa. The purpose of this study was to evaluate the properties of a new denture adhesive (Comfort-DA, developed by the authors). METHODS: Comfort-DA was evaluated in the laboratory to determine the clinical effect of pH, bonding strength, and cytotoxicity. Firstly, the material was diluted at 1.0, 2.0, 3.3, 5.0, and 10% concentrations. Then the pH values were measured at 1-, 2-, 3-, 4-, 5-, 6-, and 8-hour intervals. Secondly, bonding load test was performed between acrylic resin samples on a universal testing machine. An existing product, Fittydent, was used as a control. Finally, the cytotoxicity of several dilutions of Comfort-DA was recorded with a MTT assay on human oral fibroblast cells. Cell viability was recorded by optical density after culturing in an atmosphere of 5% CO2 and 95% air at 37 degrees C for 2, 3 and 4 days, respectively. Wells containing fresh medium alone served as controls. All the results were compared with ANOVA or Tukey intervals (alpha = 0.05). RESULTS: Comfort-DA remained above the critical pH for hydroxyapatite (pH 6.5) in all dilutions throughout the study. Results of bonding strength test indicated that Comfort-DA was significantly stronger than Fittydent (p < 0.05). The statistical analysis of results for cytotoxicity showed that Comfort-DA only displayed slight cytotoxicity. SIGNIFICANCE: The generally favorable laboratory performance of Comfort-DA formulations indicates that this product may be an efficacious denture adhesive.