RESUMEN
Lymph node metastasis (LNM) is one of the common features of oral tongue squamous cell carcinoma (OTSCC). LNM is also taken as a sign of advanced OTSCC and poor survival rate. Recently, single-cell RNA sequencing has been applied in investigating the heterogeneity of tumor microenvironment and discovering the potential biomarkers for helping the diagnosis and prognosticating. Pathogenesis of LNM in OTSCC remains unknown. Specifically, cancer-associated fibroblasts (CAFs) and epithelial tumor cells could foster the progression of tumors. Thus, in this study, we aimed to comprehensively analyze the roles of subpopulations of CAFs and epithelial tumor cells in lymph node metastatic OTSCC using the integration of OTSCC single-cell RNA sequencing datasets. Four distinct subtypes of CAFs, namely vascular CAFs, myofibroblast CAFs, inflammatory CAFs, and growth arrest CAFs were successfully discovered in LNM tumor and confirmed the roles of GAS and PTN pathways in the progression of tumor metastasis. In addition, NKAIN2+ epithelial cells and FN1+ epithelial cells specifically exhibited an upregulation of PTN, NRG, MIF, and SPP1 signaling pathways in the metastatic OTSCC. In doing so, we put forth some potential biomarkers that could be utilized for the purpose of diagnosing and prognosticating OTSCC during its metastatic phase and tried to confirm by immunofluorescence assays.
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Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Neoplasias de la Lengua , Humanos , Carcinoma de Células Escamosas de Cabeza y Cuello/patología , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Neoplasias de la Lengua/genética , Neoplasias de la Lengua/patología , Fibroblastos/patología , Células Epiteliales/patología , Biomarcadores , Metástasis Linfática/patología , Neoplasias de Cabeza y Cuello/patología , Análisis de Secuencia de ARN , Microambiente TumoralRESUMEN
BACKGROUND: Peri-implantitis is a polybacterial infection that can lead to the failure of dental implant rehabilitation. This study aimed to profile the microbiome of the peri-implant plaque and estimate the effect of periodontitis on it among 40 Chinese participants with dental implant prostheses and presenting with varying peri-implant and periodontal health states. METHODS: Submucosal plaque samples were collected from four distinct clinical categories based on both their implant and periodontal health status at sampling point. Clinical examinations of dental implant and remaining teeth were carried out. Metagenomic analysis was then performed. RESULTS: The microbiome of the peri-implantitis sites differed from that of healthy implant sites, both taxonomically and functionally. Moreover, the predominant species in peri-implantitis sites were slightly affected by the presence of periodontitis. T. forsythia, P. gingivalis, T. denticola, and P. endodontalis were consistently associated with peri-implantitis and inflammatory clinical parameters regardless of the presence of periodontitis. Prevotella spp. and P. endodontalis showed significant differences in the peri-implantitis cohorts under different periodontal conditions. The most distinguishing function between diseased and healthy implants is related to flagellar assembly, which plays an important role in epithelial cell invasion. CONCLUSIONS: The composition of the peri-implant microbiome varied in the diseased and healthy states of implants and is affected by individual periodontal conditions. Based on their correlations with clinical parameters, certain species are associated with disease and healthy implants. Flagellar assembly may play a vital role in the process of peri-implantitis.
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Implantes Dentales , Placa Dental , Microbiota , Periimplantitis , Enfermedades Periodontales , Periodontitis , Humanos , Periimplantitis/microbiología , Implantes Dentales/microbiología , Estudios Transversales , Placa Dental/microbiologíaRESUMEN
BACKGROUND: Spontaneous spheroid culture is a novel three-dimensional (3D) culture strategy for the rapid and efficient selection of progenitor cells. The objectives of this study are to investigate the pluripotency and differentiation capability of spontaneous spheroids from alveolar bone-derived mesenchymal stromal cells (AB-MSCs); compare the advantages of spontaneous spheroids to those of mechanical spheroids; and explore the mechanisms of stemness enhancement during spheroid formation from two-dimensional (2D) cultured cells. METHODS: AB-MSCs were isolated from the alveolar bones of C57BL/6 J mice. Spontaneous spheroids formed in low-adherence specific culture plates. The stemness, proliferation, and multi-differentiation capacities of spheroids and monolayer cultures were investigated by reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunofluorescence, alkaline phosphatase (ALP) activity, and oil-red O staining. The pluripotency difference between the spontaneous and mechanical spheroids was analyzed using RT-qPCR. Hypoxia-inducible factor (HIFs) inhibition experiments were performed to explore the mechanisms of stemness maintenance in AB-MSC spheroids. RESULTS: AB-MSCs successfully formed spontaneous spheroids after 24 h. AB-MSC spheroids were positive for MSC markers and pluripotency markers (Oct4, KLF4, Sox2, and cMyc). Spheroids showed higher Ki67 expression and lower Caspase3 expression at 24 h. Under the corresponding conditions, the spheroids were successfully differentiated into osteogenic and adipogenic lineages. AB-MSC spheroids can induce neural-like cells after neurogenic differentiation. Higher expression of osteogenic markers, adipogenic markers, and neurogenic markers (NF-M, NeuN, and GFAP) was found in spheroids than in the monolayer. Spontaneous spheroids exhibited higher stemness than mechanical spheroids did. HIF-1α and HIF-2α were remarkably upregulated in spheroids. After HIF-1/2α-specific inhibition, spheroid formation was significantly reduced. Moreover, the expression of the pluripotency genes was suppressed. CONCLUSIONS: Spontaneous spheroids from AB-MSCs enhance stemness and pluripotency. HIF-1/2α plays an important role in the stemness regulation of spheroids. AB-MSC spheroids exhibit excellent multi-differentiation capability, which may be a potent therapy for craniomaxillofacial tissue regeneration.
Asunto(s)
Células Madre Mesenquimatosas , Esferoides Celulares , Animales , Ratones , Técnicas de Cultivo de Célula/métodos , Ratones Endogámicos C57BL , Células Cultivadas , Células Madre , Diferenciación Celular , Osteogénesis/genética , Hipoxia/metabolismoRESUMEN
Bone defect regeneration depends on the population and lifespan of M2 macrophages, which are regulated by dual signals generated by the "physical" spatial configuration of biological tissues and "molecular" chemokines. Herein, inspired by the reprogramming of macrophages, immunoengineered porous microspheres are constructed to accelerate bone repair through the regulation of both "physical" and "molecular" signals. The porous structure of injectable poly (l-lactic acid) (PLLA) microspheres prepared by the microfluidic technique provides a "physical signal" for osteogenic differentiation. Additionally, interleukin (IL)-4-loaded liposomes (Ls) are modified on PLLA microspheres through amide bonds to produce IL-4/Ls/PLLA microspheres, providing a "molecular signal" in stimulating the differentiation of macrophages to M2 type. It is confirmed that IL-4/Ls/PLLA microspheres could induce M2-macrophages polarization and potentiate osteoblast proliferation and differentiation while coculturing with macrophages and osteoblasts in vitro. Besides, IL-4/Ls/PLLA microspheres are proved to promote bone defect regeneration by inducing the conversion of M1 macrophages to M2 through dual biosignal-functional regulation in both the calvaria defect and maxillary sinus defect models. Overall, the immuno-reprogrammed IL-4/Ls/PLLA microspheres achieve the precise immuno-reprogramming of macrophages by dual biosignal-functional regulation. This immune reengineering strategy paves a way for clinical bone defect treatment.
Asunto(s)
Interleucina-4 , Osteogénesis , Regeneración Ósea/fisiología , Microesferas , Osteoblastos , Poliésteres/químicaRESUMEN
BACKGROUND AND OBJECTIVE: Type 2 diabetes (T2D)-associated periodontitis is severe and refractory in many cases. Considered an inflammatory disease, T2D predisposes to periodontitis by increasing whole-body inflammation. One mechanism of increased inflammation is thatT2D is mediated by loss of production or function of the anti-inflammatory hormone adiponectin. In our previous report, AdipoRon, an adiponectin receptor agonist, and AdipoAI, a newly discovered, more specific agonist, attenuated T2D-associated inflammation by inhibiting osteoclastogenesis and LPS-induced endotoxemia. Autophagy plays an important role during osteoclast differentiation and function. The impact of AdipoAI on osteoclast function and autophagy involved in osteoclastogenesis is not known. Here, we compare AdipoRon and AdipoAI potency, side effects and mechanism of action in T2D-associated periodontitis. METHODS: The RAW 264.7 cell line was used for in vitro studies. We analyzed the potential cytotoxicity of AdipoAI using the CCK-8 assay. The anti-osteoclastogenic potential of AdipoAI was studied by real-time qPCR and tartrate-resistant acid phosphatase staining. The actions of AdipoAI involved in autophagy were tested by real-time qPCR, western blot and immunofluorescence staining. In the diet-induced obesity model of T2D, we investigated the impact of AdipoAI on fasting blood glucose, alveolar bone loss, and gingival inflammation in mice with experimental periodontitis. RESULTS: AdipoRon inhibited osteoclastogenesis and AdipoAI inhibited osteoclastogenesis at lower doses than AdipoRon without any cytotoxicity. In DIO mice with experimental periodontitis, AdipoAI reduced mouse body weight in 14 days, reducing fasting glucose levels, alveolar bone destruction, osteoclast number along the alveolar bone surface, and decreased the expression of pro-inflammatory factors in periodontal tissues. AdipoAI and AdipoRon also enhanced LC3A/B expression when cultured with RANKL.3-Methyladenine, a known autophagy inhibitor, decreased LC3A/B expression and reversed the inhibition of osteoclastogenesis during AdipoAI treatment. CONCLUSIONS: Our results demonstrate that AdipoAI ameliorates the severity of T2D-associated periodontitis by enhancing autophagy in osteoclasts at lower doses than AdipoRon without demonstrable side effects. Thus, AdipoAI has pharmaceutical potential for treating diabetes-associated periodontal disease.
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Pérdida de Hueso Alveolar , Diabetes Mellitus Tipo 2 , Periodontitis , Adiponectina , Pérdida de Hueso Alveolar/tratamiento farmacológico , Pérdida de Hueso Alveolar/metabolismo , Pérdida de Hueso Alveolar/prevención & control , Animales , Autofagia , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/tratamiento farmacológico , Diabetes Mellitus Tipo 2/metabolismo , Ratones , Osteoclastos , Periodontitis/tratamiento farmacológico , Periodontitis/metabolismo , Ligando RANK/metabolismo , Receptores de Adiponectina/agonistas , Receptores de Adiponectina/metabolismo , Receptores de Adiponectina/uso terapéuticoRESUMEN
Periodontitis is one of the most common chronic inflammations of the oral cavity, which eventually leads to tooth loss. Betulinic acid (BetA) is an organic acid that has anti-inflammatory effects and is derived from fruits and plants, but its effect on the osteogenic differentiation of human periodontal ligament stem cells (hPDLSCs) is still unclear. This study aimed to explore the effect of BetA on the osteogenic differentiation of hPDLSCs and its mechanism. Our results revealed that BetA not only promoted the viability of hPDLSCs but also induced their osteogenic differentiation in a dose-dependent manner. In addition, RNA sequencing was used to screen the differentially expressed genes (DEGs) after hPDLSCs were treated with BetA, and 127 upregulated and 138 downregulated genes were identified. Gene Ontology enrichment analysis showed that DEGs were mainly involved in the response to lithium ions and the positive regulation of macrophage-derived foam cell differentiation. The Kyoto Encyclopedia of Genes and Genomes analysis results revealed that DEGs were enriched in the nuclear factor-κB and interleukin-17 signaling pathways. More importantly, we confirmed that early growth response gene 1 (EGR1), one of the three DEGs involved in bone formation, significantly promoted the expression of osteogenic markers and the mineralization of hPDLSCs. Knockdown of EGR1 obviously limited the effect of BetA on the osteogenic differentiation of hPDLSCs. In conclusion, BetA promoted the osteogenic differentiation of hPDLSCs through upregulating EGR1, and BetA might be a promising candidate in the clinical application of periodontal tissue regeneration.
Asunto(s)
Diferenciación Celular/efectos de los fármacos , Proteína 1 de la Respuesta de Crecimiento Precoz/metabolismo , Osteogénesis/efectos de los fármacos , Triterpenos Pentacíclicos/farmacología , Ligamento Periodontal/efectos de los fármacos , Ligamento Periodontal/metabolismo , Células Madre/metabolismo , Adulto , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Proteína 2 de la Respuesta de Crecimiento Precoz/genética , Proteína 2 de la Respuesta de Crecimiento Precoz/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Ligamento Periodontal/citología , Proteínas Proto-Oncogénicas c-fos/genética , Proteínas Proto-Oncogénicas c-fos/metabolismo , Transducción de Señal/efectos de los fármacos , Células Madre/citología , Células Madre/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Adulto Joven , Ácido BetulínicoRESUMEN
LIM and SH3 protein 1 (LASP1) is a specific focal adhesion protein that promotes metastasis in a variety of tumours. However, its role in head and neck squamous cell carcinoma (HNSCC) has not been fully validated. The purpose of this study was to analyse the interaction of LASP1 and its binding partner in HNSCC. The expression of LASP1 and HSPA1A in HNSCC was analysed by real-time PCR and Western blot. The effects of LASP1 on the biology behaviour of HNSCC cell lines were observed in vivo and in vitro. Co-immunoprecipitation analysis was performed to confirm the interaction between LASP1 and HSPA1A. LASP1 was highly expressed in HNSCC and associated with poor prognosis for patients. LASP1 also promoted cell proliferation, colony formation, invasion and cell cycle G2/M phase transition. Heat shock protein family A member 1A (HSPA1A) is identified as a chaperone protein of LASP1 and co-localized in the cytoplasm. HSPA1A positively regulates the interaction of LASP1 with P-AKT and enhances the malignant behaviour of HNSCC cells. LASP1 and HSPA1A are both up-regulated in HNSCC, and directly binds to each other. Double inhibition of LASP1 and HSPA1A expression may be an effective method for the treatment of HNSCC.
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Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas del Citoesqueleto/metabolismo , Proteínas HSP70 de Choque Térmico/metabolismo , Neoplasias de Cabeza y Cuello/metabolismo , Neoplasias de Cabeza y Cuello/patología , Proteínas con Dominio LIM/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/patología , Proteínas Adaptadoras Transductoras de Señales/genética , Ciclo Celular/genética , Línea Celular Tumoral , Movimiento Celular/genética , Proliferación Celular/genética , Estudios de Cohortes , Proteínas del Citoesqueleto/genética , Femenino , Regulación Neoplásica de la Expresión Génica , Proteínas HSP70 de Choque Térmico/química , Proteínas HSP70 de Choque Térmico/genética , Neoplasias de Cabeza y Cuello/genética , Humanos , Proteínas con Dominio LIM/genética , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , Metástasis de la Neoplasia , Fosforilación , Pronóstico , Unión Proteica , Proteínas Proto-Oncogénicas c-akt/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello/genética , Regulación hacia Arriba/genéticaRESUMEN
Fluorosis and bone pathologies can be caused by chronic and/or excessive fluoride intake. Despite this, few studies have been conducted on the cellular mechanisms underlying osteoblast toxicity in the presence of NaF. Here, we investigated the effects of fluoride on MC3T3-E1 cells. We showed that the proliferation of MC3T3-E1 cells was inhibited by exposure to NaF. In addition, apoptosis was induced by NaF, as caspase-associated proteins showed a higher level of expression and apoptotic bodies were formed. Furthermore, endoplasmic reticulum (ER) stress induced by NaF activated the unfolded protein response (UPR) and upregulated the expression of the glucose-regulated proteins 94 (GRP94) and 78 (BiP). Therefore, ER stress plays a vital role in NaF-induced autophagy and apoptosis. Furthermore, apoptosis is promoted following the inhibition of NaF-induced autophagy. In conclusion, under NaF treatment, the ER stress-signaling pathway is activated, leading to apoptosis and autophagy and affecting the proliferation and survival of MC3T3-E1 cells.
Asunto(s)
Apoptosis , Autofagia , Estrés del Retículo Endoplásmico , Osteoblastos/efectos de los fármacos , Fluoruro de Sodio/toxicidad , Animales , Línea Celular , Ratones , Osteoblastos/fisiología , Transducción de Señal , Fluoruro de Sodio/farmacología , Respuesta de Proteína DesplegadaRESUMEN
Oral squamous cell carcinoma (OSCC) is a highly prevalent cancer worldwide, and OSCC often goes undiagnosed until advanced disease is present, which contributes to a low survival rate for OSCC patients. The identification of biomarkers for the early detection OSCC and novel therapeutic targets for OSCC treatment is an important research objective. We performed bioinformatics analyses of the gene expression profile of OSCC using microarray data to identify genes that contribute to the development of OSCC. We also predicted the transcription factors involved in the regulation of differential gene expression in OSCC. Our results showed that PI3K, EGFR, STAT1, and CPBP are important contributors to the changes in cellular physiology that occur during the development of OSCC. Therefore, these genes represent potential diagnostic biomarkers and therapeutic targets for OSCC.
RESUMEN
Satb2 is a special AT-rich binding transcription factor essential for osteoblast differentiation and bone formation. Specific microRNAs (miRNAs) have been identified to regulate the complex process of osteogenic differentiation. It remains unclear how miRNA expressions is changed in the Satb2-induced osteogenic differentiation of bone marrow stromal cells (BMSCs). From the miRNA expression profile data collected by us from Satb2-induced osteogenic differentiation of mouse BMSCs, we found that miR-27a was significantly down-regulated relative to non-treated cells 7 days post induction. By in silico analysis, we identified Sp7 as a miR-27a targeting gene and verified the findings by Western blot analysis, qRT-PCR, and luciferase reporter assays. We also analyzed the function of miR-27a in osteogenic differentiation by transfection of exogenous miR-27a into BMSCs. Overexpression of miR-27a remarkably inhibited the expression of Sp7 and attenuated Satb2-induced osteogenic differentiation. Our results suggest that expression of Sp7 during the early stage of Satb2-induced osteogenic differentiation of BMSCs is regulated by miR-27a.
Asunto(s)
Células de la Médula Ósea/metabolismo , Diferenciación Celular , Proteínas de Unión a la Región de Fijación a la Matriz/metabolismo , MicroARNs/metabolismo , Osteogénesis , Factores de Transcripción/metabolismo , Animales , Células de la Médula Ósea/citología , Ratones , Factor de Transcripción Sp7 , Células del Estroma/citología , Células del Estroma/metabolismoRESUMEN
Satb2 acts as a potent transcription factor to promote osteoblast differentiation and bone regeneration. Recently, microRNAs (miRNA) have been identified as critical regulators of osteogenic differentiation. This study aimed to identify specific miRNAs and their regulatory roles in the process of Satb2-induced osteogenic differentiation. We studied the differentially expressed miRNAs by Satb2 overexpression in murine bone marrow stromal cells using miRNA microarray. Ten down-regulated miRNAs including miR-27a, miR-125a-5p, and miR-466f-3p, and 18 up-regulated miRNAs including miR-17, miR-20a and miR-210 were found to be differentially expressed and their expression were verified by quantitative real time PCR. The differentially expressed miRNAs were further subjected to gene ontology and KEGG analysis. The highly enriched GOs and KEGG pathway showed target genes of these miRNAs were significantly involved in multiple biological processes (mesenchymal cell differentiation, bone formation, and skeletal development), and several osteogenic pathways (TGF-ß/BMP, MAPK, and Wnt signaling pathway). Finally, miR-27a was selected for target verification and function analysis. BMP2, BMPR1A, and Smad9, members of the TGF-ß/BMP superfamily, which were predicted to be target genes of miR-27a, were confirmed to be significantly up-regulated in Satb2-overexpressing cells by quantitative real time PCR. Overexpression of miR-27a significantly inhibited osteogenesis and repressed BMP2, BMPR1A, and Smad9 expression. In this study, we identified that a number of differentially regulated miRNAs, whose target genes involved in the TGF-ß/BMP signaling pathway, play an important role in the early stage of Satb2-induced osteogenic differentiation.
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Diferenciación Celular , Proteínas de Unión a la Región de Fijación a la Matriz/fisiología , Células Madre Mesenquimatosas/fisiología , MicroARNs/metabolismo , Factores de Transcripción/fisiología , Transcriptoma , Fosfatasa Alcalina/metabolismo , Animales , Proteína Morfogenética Ósea 2/genética , Proteína Morfogenética Ósea 2/metabolismo , Receptores de Proteínas Morfogenéticas Óseas de Tipo 1/genética , Receptores de Proteínas Morfogenéticas Óseas de Tipo 1/metabolismo , Células Cultivadas , Ontología de Genes , Ratones , Ratones Endogámicos C57BL , MicroARNs/genética , Osteogénesis , Interferencia de ARN , Transducción de Señal , Proteína Smad8/genética , Proteína Smad8/metabolismo , Ingeniería de Tejidos , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
Special AT-rich sequence-binding protein 2 (SATB2 ) acts as a potent transcription factor to promote osteoblast differentiation and bone regeneration. In this study, we first used lentiviral-mediated gene transfer of Satb2 into mouse bone marrow stromal cells (BMSCs) and investigated the capacity of SATB2 overexpression to promote osteogenic differentiation in vitro and in vivo. We found that LV-Satb2 -transduced BMSCs produced SATB2 protein and underwent rapid and marked osteogenic differentiation, as demonstrated by increased expression of osteoblastic genes, including runt-related transcription factor 2 (Runx2), transcription factor Sp7 (Sp7), activating transcription factor 4 (Atf4), and bone sialoprotein (Bsp), and increased alkaline phosphatase activity and Alizarin Red S staining. To analyze the induction of bone formation in vivo, LV-Satb2-transduced BMSCs were implanted into the hindlimbs of syngeneic mice, with ß-tricalcium phosphate as the scaffolding material. Four weeks after implantation, transduction with LV-Satb2 had greatly enhanced the formation of new bone. These data demonstrated the capacity of lentiviral-mediated SATB2 to promote the osteogenic differentiation of BMSCs in vitro and to enhance bone formation through a tissue-engineering technique that may be useful in bone-regenerative medicine.
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Vectores Genéticos/genética , Lentivirus/genética , Proteínas de Unión a la Región de Fijación a la Matriz/genética , Células Madre Mesenquimatosas/fisiología , Osteogénesis/genética , Factores de Transcripción/genética , Factor de Transcripción Activador 4/análisis , Fosfatasa Alcalina/análisis , Animales , Regeneración Ósea/fisiología , Fosfatos de Calcio/química , Técnicas de Cultivo de Célula , Diferenciación Celular/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/análisis , Técnicas de Transferencia de Gen , Sialoproteína de Unión a Integrina/análisis , Masculino , Trasplante de Células Madre Mesenquimatosas/métodos , Ratones , Ratones Endogámicos C57BL , Osteoblastos/fisiología , Factor de Transcripción Sp7 , Ingeniería de Tejidos/métodos , Andamios del Tejido/química , Factores de Transcripción/análisis , Trasplante Isogénico , Dedos de Zinc/genéticaRESUMEN
BACKGROUND: This study aims to observe the morphological characteristics and identify the function characteristics of junctional epithelium (JE) tissues and cultured JE cells. METHODS: Paraffin sections of human molar or premolar on the gingival buccolingual side were prepared from 6 subjects. HE staining and image analysis were performed to measure and compare the morphological difference among JE, oral gingival epithelium (OGE) and sulcular epithelium (SE). Immunohistochemistry was applied to detect the expression pattern of cytokeratin 5/6, 7, 8/18, 10/13, 16, 17, 19, and 20 in JE, OGE and SE. On the other hand, primary human JE and OGE cells were cultured in vitro. Cell identify was confirmed by histology and immunohistochemistry. In a co-culture model, TEM was used to observe the attachment formation between JE cells and tooth surface. RESULTS: Human JE was a unique tissue which was different from SE and OGE in morphology. Similarly, morphology of JE cells was also particular compared with OGE cells cultured in vitro. In addition, JE cells had a longer incubation period than OGE cells. Different expression of several CKs illustrated JE was in a characteristic of low differentiation and high regeneration. After being co-cultured for 14 d, multiple cell layers, basement membrane-like and hemidesmosome-like structures were appeared at the junction of JE cell membrane and tooth surface. CONCLUSIONS: JE is a specially stratified epithelium with low differentiation and high regeneration ability in gingival tissue both in vivo and in vitro. In co-culture model, human JE cells can form basement membrane-like and hemidesmosome-like structures in about 2 weeks.
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Inserción Epitelial/anatomía & histología , Membrana Basal/anatomía & histología , Adhesión Celular/fisiología , Técnicas de Cultivo de Célula , Diferenciación Celular/fisiología , Membrana Celular/fisiología , Membrana Celular/ultraestructura , Técnicas de Cocultivo , Inserción Epitelial/citología , Inserción Epitelial/fisiología , Células Epiteliales/citología , Células Epiteliales/fisiología , Epitelio/anatomía & histología , Epitelio/fisiología , Encía/anatomía & histología , Encía/citología , Encía/fisiología , Hemidesmosomas/ultraestructura , Humanos , Queratinas/análisis , Regeneración/fisiología , Técnicas de Cultivo de Tejidos , Raíz del Diente/anatomía & histologíaRESUMEN
OBJECTIVE: To compare the outcomes of open healing to complete closure for collagen membrane coverage for immediate implant placements with simultaneous guided bone regeneration (GBR) in two retrospective cohorts. METHODS: The subjects included 118 patients who received Bio-Gide® collagen membrane coverage for immediate implant placements and GBR in 20 anterior and 98 posterior teeth. For 58 patients, gingival flaps were released to achieve full coverage of collagen membrane (CC group). For 60 patients, no efforts were made to release the gingival flaps and collagen membrane was left exposed for open healing (OH group). Antibiotics and analgesics were prescribed for 7 days after surgery. The width of crestal open wounds were measured after surgery (W0), and at 1, 2 and 16 weeks (W16). Changes in bone mass were assessed by cone-beam computed tomography after implant placement and again at W16. Gingival and bone tissues over the implant cover screws were harvested and assessed for 16 patients in the OH group at W16. RESULTS: No wound dehiscence occurred in the CC group from W0 to W16. Both the vertical and horizontal bone dimension changes were not significantly different between the OH and CC group. For the OH group, soft tissue was completely healed at W16 when the initial wound widths were ≤6 mm. For those with initial wound widths ≥ 7 mm, the cover screws were exposed in 5/16 patients at W16 but did not affect the final restorations. Tissue staining showed keratinized mucosa and new bone formation above the dental implant in the OH group. CONCLUSION: Open healing achieved healing outcomes similar to those of complete closure for collagen membrane coverage following immediate implant placements. CLINICAL SIGNIFICANCE: For immediate implant placement requiring bone grafting and collagen membrane coverage, it is unnecessary to release the gingival flaps or use tissue grafts to achieve full coverage of the crestal wounds. Open healing with exposed membrane could achieve similar outcomes with less pain and swelling.
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Implantes Dentales , Regeneración Tisular Dirigida , Humanos , Implantación Dental Endoósea/métodos , Estudios Retrospectivos , Colágeno/uso terapéutico , Regeneración Tisular Dirigida/métodos , Regeneración ÓseaRESUMEN
The aim of this study was to evaluate the bone tissue effects under dynamic loading using finite element analysis (FEA) for four angled abutments with different deviated palatal lateral tilt angles. A three-dimensional model of the posterior maxillary region and an implant crown model were reconstructed and assembled with a three-dimensional model of the implant, angled abutment, and central screw to create a total of 10 three-dimensional finite element models tilted at 15 ∘ , 20 ∘ , 25 ∘ , and 30 ∘ in three groups, and the dynamic loads simulating oral mastication were loaded on the implant crown to analyze the equivalent stresses and strains in the peri-implant bone tissues. Under the dynamic loading, the cortical bone on the buccal side of the implant neck showed different degrees of stress concentration, and the cortical bone stress was much higher than the cancellous bone, and the strain concentration area of each model was located in the bone tissue around the implant neck and base. For the use of angular abutment, under the premise that the cortical bone stresses and strains of the 10 models meet the requirements for use, the peak stresses of 2.907 MPa, 3.018 MPa, and 2.164 MPa were achieved by using the 20 ∘ angular abutment to achieve the tilt angles of 20 ∘ , 25 ∘ , and 30 ∘ implantation, which is more advantageous compared with other models.
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Pilares Dentales , Implantes Dentales , Análisis de Elementos Finitos , Maxilar , Estrés Mecánico , Humanos , Maxilar/fisiología , Imagenología Tridimensional/métodos , Análisis del Estrés Dental/métodos , Hueso Cortical/fisiologíaRESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Rhizoma Drynariae, as the dried rhizome of Drynaria fortunei (Kunze ex Mett.) J. Sm., is a traditional Chinese medicine for treating the injury and bone broken of falling and beating. Total flavonoids is considered as the major and effective compounds for the therapeutic efficacy of Rhizoma Drynariae. AIM OF THE STUDY: To explore the effect of total flavonoids from Rhizoma Drynariae (TFRD) on bone regeneration and the underlying mechanisms. MATERIALS AND METHODS: The effect of TFRD in various doses on bone reconstruction in cranial bone defect rats was explored in vivo. The active ingredients in TFRD-medicated serum were characterized by serum pharmacochemistry and integrated by network pharmacology analysis and target prediction. To elucidate the underlying mechanism of TFRD on bone regeneration, experimental validation in vitro was executed to assess the influence of different concentrations of TFRD-medicated serum on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). RESULTS: Micro-CT, histological examination, immunohistochemical analysis, and ELSA demonstrated that administration of TFRD could promote bone reconstruction in a rat cranial defect model. We identified 27 active components of TFRD using ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS). Results from CCK8, ALP, and Alizarin Red S staining revealed that TFRD-medicated serum notably enhanced BMSCs proliferation and osteogenic differentiation. qRT-PCR and Western blot harvested results consistent with those predicted by network pharmacology, providing further evidence that TFRD activated the TGF-ß signaling pathway to benefit bone regeneration. CONCLUSION: The active components of TFRD modulate the TGF-ß signaling pathway to facilitate osteogenesis, thereby repairing cranial bone defects.
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Osteogénesis , Polypodiaceae , Animales , Ratas , Farmacología en Red , Rizoma , Espectrometría de Masas en Tándem , Regeneración Ósea , Flavonoides/farmacología , Flavonoides/uso terapéutico , Factor de Crecimiento Transformador betaRESUMEN
Osseointegrated dental implants replace missing teeth and create an artificial surface for biofilms of complex microbial communities to grow. These biofilms on implants and dental surfaces can trigger infection and inflammation in the surrounding tissue. This study investigated the microbial characteristics of peri-implant mucositis (PM) and explored the correlation between microbial ecological imbalance, community function, and disease severity by comparing the submucosal microflora from PM with those of healthy inter-subject implants and intra-subject gingivitis (G) within a group of 32 individuals. We analyzed submucosal plaques from PM, healthy implant (HI), and G sites using metagenome shotgun sequencing. The bacterial diversity of HIs was higher than that of PM, according to the Simpson index. Beta diversity revealed differences in taxonomic and functional compositions across the groups. Linear discriminant analysis of the effect size identified 15 genera and 37 species as biomarkers that distinguished PM from HIs. Pathways involving cell motility and protein processing in the endoplasmic reticulum were upregulated in PM, while pathways related to the metabolism of cofactors and vitamins were downregulated. Microbial dysbiosis correlated positively with the severity of clinical inflammation measured by the sulcus bleeding index (SBI) in PM. Prevotella and protein processing in the endoplasmic reticulum also correlated positively with the SBI. Our study revealed PM's microbiological and functional traits and suggested the importance of certain functions in disease severity.IMPORTANCEPeri-implant mucositis is an early stage in the progression of peri-implantitis. The high prevalence of it has been a threat to the widespread use of implant prosthodontics. The link between the submucosal microbiome and peri-implant mucositis was demonstrated previously. Nevertheless, the taxonomic and functional composition of the peri-implant mucositis microbiome remains controversial. In this study, we comprehensively characterize the microbial signature of peri-implant mucositis and for the first time, we investigate the correlations between microbial dysbiosis, functional potential, and disease severity. With the help of metagenomic sequencing, we find the positive correlations between microbial dysbiosis, genus Prevotella, pathway of protein processing in the endoplasmic reticulum, and more severe mucosal bleeding in the peri-implant mucositis. Our studies offer insight into the pathogenesis of peri-implant mucositis by providing information on the relationships between community function and disease severity.
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Bacterias , Implantes Dentales , Disbiosis , Microbiota , Humanos , Masculino , Persona de Mediana Edad , Femenino , Bacterias/clasificación , Bacterias/aislamiento & purificación , Bacterias/genética , Implantes Dentales/microbiología , Implantes Dentales/efectos adversos , Adulto , Disbiosis/microbiología , Índice de Severidad de la Enfermedad , Anciano , Gingivitis/microbiología , Periimplantitis/microbiología , Mucositis/microbiología , Estomatitis/microbiología , Estomatitis/etiología , Metagenoma , Biopelículas/crecimiento & desarrolloRESUMEN
PURPOSE: This study aimed to evaluate the stress distribution and secondary stability involved in five implant strategies, including implant-supported prostheses (ISP) and tooth-implant-supported prostheses (TISP), used for bone atrophy in the maxillary posterior region with teeth loss using finite element analysis, and to explore the more desirable implant methods. METHODS: Five implant strategies were made to analyze and compare: M1, implant-supported prosthesis consisting of a short implant with a regular implant; M2, implant-supported prosthesis consisting of a tilted implant with a regular implant; M3, cantilever structure; M4, tooth-implant-supported prosthesis consisting of a short implant with a regular implant; M5, tooth-implant-supported prosthesis consisting of a regular implant, and M6, with only the natural teeth as a control group. Dynamic loading of the above models was performed in finite element analysis software to assess the stress distribution of the bone tissue and implants using the von Mise criterion. Finally, the secondary stability of different models was evaluated by modal analysis. RESULTS: The maximum stress distribution in the cortical bone in M1(60 MPa) was smaller than that in M2(97 MPa) and M3(101 MPa), The first principal strain minimum was obtained in M2 (2271µÎµ). M4 (33 MPa, 10085 Hz) with the best mechanical properties and highest resonance frequency. But increased the loading on the natural teeth. CONCLUSIONS: Short implants and tilted implants are both preferred implant strategies, if cantilever construction is necessary, a tooth-implant-supported prosthesis consisting of a short implant and a regular implant is recommended.
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OBJECTIVES: This study aimed to explore the influence of alveolar bone morphologic variables on the outcome of guided bone regeneration (GBR) in the anterior maxilla region. METHODS: Twenty-eight patients who received single maxillary anterior tooth delayed implant placed simultaneously with GBR were recruited. Baseline data including age, gender, implant site, implant brand, and bone graft materials were recorded. The resorption rate of the grafted bone (RRGB), labial bone width at 0 mm, 2 mm, and 4 mm apical to the implant platform at Tn (LBW0Tn, LBW2Tn, LBW4Tn), implant angulation (IA), maximum bone graft thickness (MBGT), bone graft volume (BGV), and the initial bone morphologic variables bone concavity depth (BCD) and bone concavity angulation (BCA) were measured. The Pearson correlation analysis, analysis of variance (ANOVA), and optimal binning method were used to explore the potential predictors for GBR. RESULTS: Among 28 patients, the labial bone width of implant and bone graft volume decreased significantly when measured 6 months after surgery. The mean percentage of RRGB was 49.78%. RRGB was not correlated with gender, age, bone graft material, IA, MBGT, bone graft volume at T1, implant site, and implant brand (P > .05). BCD and BCA were each moderately correlated with RRGB (r = -0.872 [P < .001] and r = 0.686 [P < .001], respectively). A BCD ≥1.03 mm and a BCA <155.30° resulted in a significantly lower percentage of RRGB (P < .001). CONCLUSIONS: A significant grafted bone materials volume reduction was detected after GBR with collagen membrane and deproteinized bovine bone mineral (DBBM). The initial bone morphology can influence GBR outcome, and a bone concavity with a depth ≥1.03 mm and an angulation <155.30° led to a lower RRGB. BCD and BCA can be used as variables to predict the outcome of GBR.
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Aumento de la Cresta Alveolar , Implantes Dentales , Humanos , Animales , Bovinos , Maxilar/cirugía , Aumento de la Cresta Alveolar/métodos , Regeneración Ósea , Colágeno , Trasplante Óseo/métodosRESUMEN
The genome of oral squamous cell carcinoma (OSCC) has been extensively characterized via bulk sequencing, revealing a multitude of genetic changes. The gene IGF2BP3, which encodes for the insulin-like growth factor 2 mRNA-binding protein 3, has been observed to be highly expressed in several types of cancer. This finding suggests that IGF2BP3 may play a significant role in the initiation and advancement of cancer. Nevertheless, the mechanisms by which IGF2BP3 contribute to OSCC are yet to be fully understood. In this study, we have observed that IGF2BP3 exhibits overexpression in OSCC. Based on our findings from bulk sequencing analysis, we have concluded that IGF2BP3 could potentially serve as a biomarker for predicting poor prognosis in OSCC. Moreover, it has been demonstrated that IGF2BP3 exhibits a significant association with the initiation and advancement of tumors both in vivo and in vitro. The evaluation of IGF2BP3 expression levels in relation to the cell cycle stage was conducted using single-cell RNA sequencing data. Tumor cells characterized by elevated IGF2BP3 expression demonstrated a higher percentage of cells in the G2/M transition phase. This study presents new findings indicating that the molecular target IGF2BP3 can serve as a prognostic indicator for tumors and has an impact on the development and progression of OSCC by influencing the regulation of the cell cycle.