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BACKGROUND: The aim of this study was to investigate the impact of Porphyromonas gingivalis (P. gingivalis) on the progression of oral squamous cell carcinoma (OSCC) through neutrophil extracellular traps (NETs) in the tumor immune microenvironment. METHODS: The expression of NETs-related markers was identified through immunohistochemistry, immunofluorescence, and Western blotting in different clinical stages of OSCC samples. The relationship between NETs-related markers and clinicopathological characteristics in 180 samples was analyzed using immunohistochemistry data. Furthermore, the ability to predict the prognosis of OSCC patients was determined by ROC curve analysis and survival analysis. The effect of P. gingivalis on the release of NETs was identified through immunofluorescence and immunohistochemistry, both in vitro and in vivo. CAL27 and SCC25 cell lines were subjected to NETs stimulation to elucidate the influence of NETs on various cellular processes, including cell proliferation, migration, invasion, and metastasis in vitro. Furthermore, the impact of NETs on the growth and metastatic potential of OSCC was assessed using in vivo models involving tumor-bearing mice and tumor metastasis mouse models. RESULTS: Immunochemistry analysis revealed a significant correlation between the NETs-related markers and clinical stage, living status as well as TN stage. P. gingivalis has demonstrated its ability to effectively induce the release of NETs both in vivo and in vitro. NETs have the potential to facilitate cell migration, invasion, and colony formation. Moreover, in vivo experiments have demonstrated that NETs play a pivotal role in promoting tumor metastasis. CONCLUSION: High expression of NETs-related markers demonstrates a strong correlation with the progression of OSCC. Inhibition of the NETs release process stimulated by P. gingivalis and targeted NETs could potentially open up a novel avenue in the field of immunotherapy for patients afflicted with OSCC.
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Carcinoma de Células Escamosas , Armadilhas Extracelulares , Neoplasias Bucais , Porphyromonas gingivalis , Microambiente Tumoral , Porphyromonas gingivalis/imunologia , Humanos , Armadilhas Extracelulares/imunologia , Armadilhas Extracelulares/metabolismo , Microambiente Tumoral/imunologia , Animais , Neoplasias Bucais/imunologia , Neoplasias Bucais/patologia , Neoplasias Bucais/microbiologia , Linhagem Celular Tumoral , Feminino , Masculino , Carcinoma de Células Escamosas/imunologia , Carcinoma de Células Escamosas/patologia , Pessoa de Meia-Idade , Camundongos , Progressão da Doença , Camundongos Endogâmicos BALB C , Proliferação de Células , Movimento Celular , Camundongos Nus , Infecções por Bacteroidaceae/imunologia , Infecções por Bacteroidaceae/microbiologia , Neutrófilos/imunologia , IdosoRESUMO
OBJECTIVE: The aim of this study was to investigate the effect of mammalian-enabled (Mena) on tongue squamous cell carcinoma (TSCC) metastasis and its mechanism. MATERIALS AND METHODS: Immunochemistry was performed to investigate the Mena and tumor-related markers expression, and its clinicopathological characteristics in 46 TSCC specimens. TSCC cell SCC9 and Cal27 untransfected or stable transfected with Mena overexpression and small interfering RNA were used to determine the role of Mena in cell proliferation, cell migration, invasion and metastasis, and EMT-related markers in vitro, and the effect of Mena on TSCC growth and metastasis through tumor-bearing and tumor metastasis immunodeficient mice models in vivo. RESULTS: Immunochemistry showed that the expression of Mena was significantly correlated with lymphatic metastasis and TNM stage, E-cadherin, Vimentin, and MMP2. Mena did not affect cell proliferation and colony formation in vitro, and tumor growth in vivo. However, it promoted cell migration and invasion in vitro, and TSCC metastasis in vivo. CONCLUSIONS: Mena expression is associated with lymphatic metastasis and tumor stage and promotes TSCC invasion and metastasis by inducing the EMT process. Thus, Mena may be a biomarker for prognosis and targeted therapy in TSCC patients.
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This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editor-in-Chief. The authors have duplicated an image that previously appeared in: J Clin Otorhinolaryngol Head Neck Surg (China) 2018 https://doi.org/10.13201/j.issn.1001-1781.2018.16.011 One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents a severe abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of JOMS that this was not detected during the submission process.
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BACKGROUND: The surgical extraction of impacted third molars is one of the most common procedures in oral and maxillofacial surgery, which associated with several postoperative complications. The aim of this clinical trial was to estimate the implication of concentrated growth factor (CGF) on postoperative sequelae after the completely impacted lower third molar extraction. MATERIALS AND METHODS: A total of 74 sides of 37 participants who had completely bilateral impacted lower third molars were enrolled in this split-mouth, randomized singleblind, clinical trial. Surgical extraction was undertaken on both sides of the mandible. Randomization was achieved by opaque, sealed envelopes. The postoperative outcomes including wound healing, swelling and pain were clinically assessed at different-time intervals(1st, 3rd and 7th days). A p-value < 0.05 was considered statistically significant. RESULTS: The wound healing index was significantly better in the test sides (P = 0.001). Regarding the facial swelling, the test sides had significantly less values than the control sides, particularly on the 1st (1.01 ± .57 vs. 1.55 ± .56) and 3rd days (1.42 ± 0.8 vs. 2.63 ± 1.2) postoperatively. Nonetheless, the swelling was disappeared within the 7th day in both sides. The pain scores of visual analog scale were no a statistically significant difference between both sides on the 1st day, meanwhile, the pain scores were significantly lower in the test sides compared with the control sides, especially on the 3rd (P = 0.001) and 7th days (P < 0.001) postoperatively. CONCLUSION: The application of CGF following the surgical extraction of lower third molar has accelerated the healing of soft tissues as well as reduced postoperative sequelae such as swelling and pain. Therefore, the CGF could be promoted among clinicians during the lower third molar surgical extraction. TRIAL REGISTRATION: This study was registered with the TCTR identification number TCTR20210325002 on 25/03/2021 at Thai Clinical Trials Register-Medical Research Foundation of Thailand (MRF). Also it was ethically approved from the institutional ethics committee at the Hospital of Stomatology, Xian Jiaotong University, Xian, China (No: 032), and has been conducted in accordance to the guidelines of the declaration of Helsinki. Written informed consent was obtained from all participants in the study.
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Dente Serotino , Dente Impactado , Edema/etiologia , Edema/prevenção & controle , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Mandíbula/cirurgia , Dente Serotino/cirurgia , Dor/complicações , Dor Pós-Operatória/etiologia , Dor Pós-Operatória/prevenção & controle , Método Simples-Cego , Extração Dentária/efeitos adversos , Dente Impactado/cirurgiaRESUMO
PURPOSE: To evaluate the usefulness of the modified cosmetic incision (MCI) and advanced wound closure method in partial parotidectomy by comparison with the modified Blair incision (MBI). PATIENTS AND METHODS: This study retrospectively enrolled 44 patients who underwent partial parotidectomy for benign parotid tumors. These patients were divided into 2 groups: MCI group and MBI group. The MCI surgical procedures were performed via a minimal facelift incision with no preauricular incision, postauricular and hairline incision, or extensive hairline incision and an advanced wound closure method, using continuous absorbable intradermal sutures and skin adhesive. The MBI surgical procedures were performed via a conventional MBI and standard transdermal, interrupted, nonabsorbable suturing approach. The operation variables and the cosmetic results of the patients in each group were compared. RESULTS: A total of 23 patients underwent the MCI and advanced wound closure approach and 21 patients underwent the MBI and standard wound closure approach. No significant differences were found in gender, mean age, tumor size, or tumor site between the 2 groups (P > .05). No differences between groups were seen in operative time and intraoperative blood loss volume (P > .05). Several postoperative complications, such as facial paralysis, Frey syndrome, salivary fistula, infection, or tumor recurrence, did not differ between the 2 groups (P > .05). However, postoperative drainage volume in the MCI group was significantly lower than that in the MBI group (P < .01). Moreover, the postoperative cosmetic satisfaction, skin numbness, and scar evaluation results in the MCI group were better than those in the MBI group (P < .001). CONCLUSIONS: MCI combined with continuous absorbable intradermal sutures and skin adhesive for partial parotidectomy is technically feasible and safe and could produce excellent cosmetic outcomes in selected patients with benign parotid tumors.
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Cicatriz , Neoplasias Parotídeas , Dissecação , Humanos , Recidiva Local de Neoplasia , Neoplasias Parotídeas/cirurgia , Estudos RetrospectivosRESUMO
Increasing evidence has shown that aberrant miRNAs contribute to the development and progression of human melanoma. Previous studies have shown that miR-125b functions as a suppressor in malignant melanoma. However, the molecular function and mechanism by which miR-125b influences melanoma growth and invasion are still unclear. In this study, we aimed to investigate the role of miR-125b in melanoma progression and metastasis. We found that miR-125b expression is significantly downregulated in primary melanoma, and an even greater downregulation was observed in metastatic invasion. Restored expression of miR-125b in melanoma suppressed cell proliferation and invasion both in vitro and in vivo. Furthermore, our findings demonstrate that upregulating miR-125b significantly inhibits malignant phenotypes by repressing the expression of integrin alpha9 (ITGA9). Finally, our data reveal that upregulated expression of ITGA9 in melanoma tissues is inversely associated with miR-125b levels. Together, our results demonstrate that upregulation of ITGA9 in response to the decrease in miR-125b in metastatic melanoma is responsible for melanoma tumor cell migration and invasion.
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Transição Epitelial-Mesenquimal/genética , Integrinas/genética , Melanoma/genética , Melanoma/patologia , MicroRNAs/genética , Interferência de RNA , Animais , Sequência de Bases , Sítios de Ligação , Adesão Celular/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Integrinas/química , Camundongos , MicroRNAs/química , Metástase Neoplásica , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
Objectives: Head and neck squamous cell carcinoma (HNSCC) is the most common malignancy of the head and neck. However, the molecular mechanisms governing the development of HNSCC have not been fully elucidated. Materials and Methods: Differentially expressed genes (DEGs) were screened out from The Cancer Genome Atlas (TCGA) and GSE23036 datasets. Weighted gene coexpression network analysis (WGCNA) was used to reveal the correlations among genes and to search for significantly correlated gene modules. The expression levels of genes in HNSCC and normal samples according to antibody-based detected methods was assessed by utilizing the Human Protein Atlas (HPA). The impact of the selected hub genes on the prognosis of HNSCC patients was assessed by analysing immunohistochemistry (IHC) and immunofluorescence (IF) expression levels and clinical data. Results: Twenty-four genes positively correlated with tumour status and 15 genes negatively correlated with tumour status were screened out by WGCNA. PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC and were finally screened out and verified by GEPIA and HPA database analysis. Immunohistochemistry of samples collected from 175 patients with HNSCC and subsequent statistical analysis also showed that PLAU and LAMC2 were associated with a poor prognosis in patients with HNSCC, and the levels of these two factors were positively correlated. The expression and co-localization of PLAU and LAMC2 in HNSCC tissues were confirmed by double immunofluorescence labeling. Conclusions: There was a positive correlation between PLAU and LAMC2 expression in HNSCC samples, and PLAU and LAMC2 might be independent prognostic biomarkers for HNSCC.
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BACKGROUND: Clinical instructional strategies and the climate in which teaching and learning take place have a significant impact on the quality of dental education. Therefore, this study aimed to evaluate the impact of early microsurgery training on the skills of dental intern students who are planning to join an oral and maxillofacial surgical field (DIS) as compared with junior residents within an oral and maxillofacial surgery department who had no microsurgery experience (JR). METHODS: A total of 100 trainees, 70 were DIS, while the other 30 were JR. The average age was 23.87 ± 2.05 years for DIS group and 31.05 ± 3.06 for JR group. All trainees attended a microsurgical course (theoretical and practical parts) for seven days within a Microvascular Laboratory for Research and Education of a university-affiliated tertiary hospital. Two blinded examiners had assessed the performance of trainees independently using a specific scoring system. The independent sample t-test was used to compare the effect of microsurgery training between DIS and JR groups. The significance level was set at 0.05. RESULTS: The DIS group had showed higher attendance rate than JR group (p < 0.01), with a lower absence score in DIS than JR groups (0.33 ± 0.58 vs. 2.47 ± 1.36). The total score of the theoretical test was significantly different between both groups (p < 0.01). In this context, the DIS group had revealed higher total score than JR group (15.06 ± 1.92 vs. 12.73 ± 2.49). In term of tissue preservation, there was a significant difference between both groups, with the DIS had better performance score than JR (1.49 ± 0.51 vs. 0.93 ± 0.59). Further, the practical exam score was significantly higher in DIS group than JR group (p < 0.01). CONCLUSION: Overall, the performance of dental intern students was favourably compared with junior residents in most aspects. Therefore, it is promising and essential for dental colleges to add a microsurgery course to the curriculum of dental intern students who plan to specialize in oral and maxillofacial surgery.
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Internato e Residência , Humanos , Adulto Jovem , Adulto , Estudos Prospectivos , Competência Clínica , Currículo , EstudantesRESUMO
Aim: The aim of this clinical trial was to assess the impact of autologous concentrated growth factor (CGF) as a socket-filling material and its ridge preservation properties following the lower third molar extraction. Materials and methods: A total of 60 sides of 30 participants who had completely symmetrical bilateral impacted lower third molars were enrolled. The primary outcome variables of the study were bone height and width, bone density, and socket surface area in the coronal section. Cone beam computed tomography images were obtained immediately after surgery and three months after surgery as a temporal measure. Follow-up data were compared to the baseline using paired and unpaired t-tests. Results: CGF sites had higher values in height and width when compared to control sites (Buccal wall 32.9 ± 3.5 vs 29.4 ± 4.3 mm, Lingual wall 25.4 ± 3.5 vs 23.1 ± 4 mm, and Alveolar bone width 21.07 ± 1.55vs19.53 ± 1.90 mm, respectively). Bone density showed significantly higher values in CGF sites than in control sites (Coronal half 200 ± 127.3 vs -84.1 ± 121.3 and Apical half 406.5 ± 103 vs 64.2 ± 158.6, respectively). There was a significant difference between both sites in the reduction of the periodontal pockets. Conclusion: CGF application following surgical extraction provides an easy, low-cost, and efficient option for alveolar ridge preservation. Thus, the use of CGF by dentists during dental extractions may be encouraged, particularly when alveolar ridge preservation is required. Clinical trial registration: TCTR identification, TCTR20221028003.
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Extração Dentária , Alvéolo Dental , Humanos , Tomografia Computadorizada de Feixe Cônico , Extração Dentária/efeitos adversos , Alvéolo Dental/diagnóstico por imagem , Alvéolo Dental/cirurgiaRESUMO
Objective: The effect of miR-626 on the radiosensitivity to oral squamous cell carcinoma (OSCC) was evaluated in this study. Materials and Methods: The level of miR-626 in OSCC patients was determined by analyzing the data of miRNA microarray GSE113956. miR-626 was overexpressed by miR-626 mimics and knockdown were performed by miR-626 inhibitor. The level of miR-626 was detected by quantitative real-time polymerase chain reaction. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and colony formation assays were used to detect the effect of miR-626 on the growth of OSCC cells. Flow cytometry was used to detect the apoptosis of OSCC cells. Western blot and dual luciferase reporter assays were used to explore the underlying mechanism of miR-626 regulating the radiosensitivity to OSCC. The effect of miR-626 on the radiosensitivity to OSCC were examined in an in vivo xenograft model. Results: The serum miR-626 level of OSCC patients was significantly higher than that of healthy controls. miR-626 mimics significantly promoted the OSCC cell growth, but the miR-626 inhibitor significantly suppressed the OSCC cell growth. Radiation combined with the miR-626 inhibitor significantly suppressed the cell proliferation and promoted the apoptosis of SCC-4 and HSC4 cells. Moreover, miR-626 regulates the nuclear factor kappa-B (NF-κB) signaling mediated by TRAF-interacting protein with forkhead-associated domain B. Furthermore, inhibition of miR-626 enhances the radiosensitivity to OSCC in nude mice. Conclusions: miR-626 inhibition enhanced the radiosensitivity to OSCC through the downregulation of NF-κB signaling.
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Background: Mena, a cytoskeletal regulatory protein, is involved in actin-based regulation of cell motility and adhesion, and contributes to tumor invasion and metastasis. However, the role of Mena in oral squamous cell carcinoma remains unclear. This is the first research focusing on the prognostic value of Mena in OSCC. In this study, we aimed to investigate the correlation between Mena expression and clinicopathological significance, as well as prognostic value in OSCC. Methods: Mena gene expression profiles of OSCC and normal tissues were collected from Oncomine, TCGA, and GEO databases. Biological function was analyzed through GO, KEGG and GSEA enrichment. Further, the expression level of Mena and tumor-related markers in 151 OSCC specimens was examined by IHC staining based on tissue microarray. Kaplan-Meier analysis was used to assess the prognostic performance of Mena in OSCC. Result: Mena was generally upregulation in various malignancies, especially OSCC. The functional analyses indicated that Mena was involved in the assembly and regulation of actin, cell movement, and EMT. IHC staining revealed that high expression of Mena in OSCC was correlated with Lymphatic metastasis, TNM stage, E-cadherin, Vimentin, and MMP-2, but insignificantly Ki67. Kaplan-Meier analysis demonstrated that elevated expression of Mena was significantly associated with poor overall survival and disease-free survival of OSCC patients. Conclusion: Mena could be a novel biomarker for predicting the prognosis of OSCC patients, which supports a theoretical basis for developing molecular target therapy.
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Neutrophil elastase (NE) plays a pivotal role in inflammation. However, the mechanism underlying NE-mediated inflammation in obesity remains unclear. Here, we report that NE activates protease-activated receptor-2 (PAR2), stimulates actin filament (F-actin) formation, decreases intercellular junction molecule VE-cadherin expression, and increases the permeability of human arterial endothelial cells (hECs). NE also prompts degradation of VE-cadherin and its binding proteins p120- and ß-catenins via MG132-sensitive proteasomes. NE stimulates phosphorylation of myosin light-chain (MLC) and its regulator myosin phosphatase target subunit-1 (MYPT1), a target of Rho kinase (ROCK). Inhibitors of PAR2 and ROCK prohibit NE-induced F-actin formation, MLC phosphorylation, and VE-cadherin reduction in hECs, and impede monocyte transmigration through hEC monolayer pretreated with either neutrophils or NE. Further, administration of an NE inhibitor GW311616A significantly attenuates vascular leakage, leukocyte infiltration, and the expression of proinflammatory cytokines in the white adipose tissue from high-fat diet (HFD)-induced obese mice. Likewise, NE-deficient mice are resistant to HFD-induced vascular leakage in the heart. Together, NE regulates actomyosin cytoskeleton activity and VE-cadherin expression by activating PAR2 signaling in the endothelial cells, leading to increased vascular permeability and leukocyte extravasation. Hence, inhibition of NE is a potential approach to mitigate vascular injury and leukocyte infiltration in obesity-related systemic inflammation.
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Permeabilidade Capilar , Elastase de Leucócito , Actinas/metabolismo , Animais , Células Endoteliais/metabolismo , Humanos , Inflamação/metabolismo , Elastase de Leucócito/metabolismo , Leucócitos/metabolismo , Camundongos , Camundongos Obesos , Obesidade/metabolismoRESUMO
Melanoma is an invasive and malignant type of tumor with unsatisfactory therapeutic outcomes. The present study aimed to detect the expression levels of microRNA (miR)-125b in formalin-fixed paraffin-embedded (FFPE) melanoma tissues and the association of its expression levels with the clinical features, diagnosis and prognosis of melanoma. Expression levels of miR-125b in 29 FFPE melanoma specimens (16 primary and 13 metastatic tumors), and 16 intradermal nevus (IDN) specimens as a control, were detected by reverse transcription-quantitative PCR. Associations among miR-125b expression and mortality, patient age and sex, tumor location and size, lymph node metastasis (LNM) and TNM stage were analyzed by t-test. The diagnostic value of miR-125b for melanoma was evaluated by receiver operating characteristic (ROC) curve analysis. Prognosis of patients in the microRNA-125b low- and high-expression groups was analyzed by Fisher's exact test. The association between miR-125b expression and the overall survival of patients with melanoma was assessed using Kaplan-Meier curve analysis and a Cox proportional hazards model. The results revealed that the expression levels of miR-125b in primary and metastatic melanomas were significantly lower than those in the IDN control group (P<0.05), and the expression levels of miR-125b in the metastatic group were significantly lower than those in the primary group (P<0.05). In addition, the expression levels of miR-125b were significantly associated with LNM (P=0.001) and TNM stage (P=0.004), but not with age, sex, tumor size or location (P>0.05). ROC curve analysis revealed that the area under the curve (AUC) was 0.880, with a 95% CI of 0.777-0.984 (P<0.05). The overall survival rate of the patients with a low expression level of miR-125b (20.0%) was lower than that of patients with a high expression level of miR-125b (64.3%) (P<0.05). miR-125b expression was an independent predictor of overall survival in patients with melanoma [hazard ratio (HR), 0.252; 95% CI, 0.087-0.729]. Overall, these findings indicated that a low expression level of miR-125b was associated with higher LNM and TNM stage in patients with melanoma, and that this has a certain diagnostic value. miR-125b may be used for the early screening of melanoma and determining the prognosis of patients with melanoma, and may be a potential target for the treatment of the disease.
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Pathological scarring is a result of the hypertrophy of scar tissue during tissue repair following trauma. The aim of the present study was to assess the effect of ubiquitinspecific protease 4 (USP4) silencing on pathological scarring, and to evaluate the mechanistic basis for the effect. An MTT assay was used to assess cell viability. Immunoprecipitation (IP) was used to determine ubiquitination levels of the TGFß receptor (TßR)I and Smad7. Tumor formation was assessed by injecting keloid fibroblasts. Hematoxylin and eosin staining was used to detect pathological changes in tumor tissue. Reverse transcription quantitative polymerase chain reaction and western blot analysis assays were used to evaluate the expression levels of TßRI and Smad7. Compared with the untreated control animals, cell viability and the expression of TßRI and Smad7 increased significantly in animals treated with TGFß. Short hairpin RNA for USP4 (shUSP4) decreased the cell viability of negative control cells, TGFßinduced cellular proliferation, and the expression of TßRI and Smad7. IP experiments indicated that the ubiquitination level of TßRI was decreased following USP4 silencing. There was no remarkable difference in the structure of scar tissue among the various animal groups at 14 days following treatment, while the necrotic area of the scar tissue in the shUSP4 and vialinin A (USP inhibitor)treated animals increased significantly at the 28th and 42nd day compared with the control animals. At days 14, 28 and 42, the expression levels of TßRI and Smad7 in the shUSP4 and vialinin Atreated animals were significantly decreased compared with the control animals (P<0.05). In summary, interference with or inhibition of USP4 prevented the activity of the TGFß/Smad pathway signaling and inhibited the formation of pathological scars.
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Cicatriz/genética , Queloide/genética , Receptor do Fator de Crescimento Transformador beta Tipo I/genética , Proteína Smad7/genética , Fator de Crescimento Transformador beta/genética , Proteases Específicas de Ubiquitina/genética , Animais , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Cicatriz/metabolismo , Cicatriz/patologia , Cicatriz/prevenção & controle , Fibroblastos/metabolismo , Fibroblastos/patologia , Fibroblastos/transplante , Regulação da Expressão Gênica , Humanos , Queloide/metabolismo , Queloide/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Receptor do Fator de Crescimento Transformador beta Tipo I/metabolismo , Transdução de Sinais , Proteína Smad7/metabolismo , Compostos de Terfenil/farmacologia , Fator de Crescimento Transformador beta/metabolismo , Transplante Heterólogo , Proteases Específicas de Ubiquitina/antagonistas & inibidores , Proteases Específicas de Ubiquitina/metabolismoRESUMO
Dental pulp/dentine complex regeneration is indispensable to the construction of biotissue-engineered tooth roots and represents a promising approach to therapy for irreversible pulpitis. We used a tissue-engineering method based on odontogenic stem cells to design a three-dimensional (3D) and scaffold-free stem-cell sheet-derived pellet (CSDP) with the necessary physical and biological properties. Stem cells were isolated and identified and stem cells from root apical papilla (SCAPs)-based CSDPs were then fabricated and examined. Compact cell aggregates containing a high proportion of extracellular matrix (ECM) components were observed, and the CSDP culture time was prolonged. The expression of alkaline phosphatase (ALP), dentine sialoprotein (DSPP), bone sialoprotein (BSP) and runt-related gene 2 (RUNX2) mRNA was higher in CSDPs than in cell sheets (CSs), indicating that CSDPs have greater odonto/osteogenic potential. To further investigate this hypothesis, CSDPs and CSs were inserted into human treated dentine matrix fragments (hTDMFs) and transplanted into the subcutaneous space in the backs of immunodeficient mice, where they were cultured in vivo for 6 weeks. The root space with CSDPs was filled entirely with a dental pulp-like tissue with well-established vascularity, and a continuous layer of dentine-like tissue was deposited onto the existing dentine. A layer of odontoblast-like cells was found to express DSPP, ALP and BSP, and human mitochondria lined the surface of the newly formed dentine-like tissue. These results clearly indicate that SCAP-CSDPs with a mount of endogenous ECM have a strong capacity to form a heterotopic dental pulp/dentine complex in empty root canals; this method can be used in the fabrication of bioengineered dental roots and also provides an alternative treatment approach for pulp disease.
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Polpa Dentária/fisiologia , Dentina/fisiologia , Regeneração , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Animais , Diferenciação Celular , Separação Celular , Humanos , Camundongos , Células-Tronco/citologiaRESUMO
This study investigated the hypothesis that a mesenchymal stem cells (MSC)-implant complex could be used in type 2 diabetic rats. Diabetes was modeled with type 2 diabetic rats induced by high fat diet with low dose streptozotocin (STZ) injected intraperitoneally. MSC sheets were harvested from culture flasks, wrapped around implants to construct the complexes, and then cultured in an osteogenic medium. The layered cell sheets integrated well with the implants and remained viable, with small mineralized nodules visible on the implant surfaces after culturing. The MSC-implant complexes were inserted into the right tibiae of the diabetic rats. Titanium implants served as controls. After four and eight weeks of healing, the tibiae were observed via MicroCT and harvested for histological examination. For the MSC-implant complexes, MicroCT analysis showed that bone volume ratio and trabecular thickness increased significantly (p<0.05), and trabecular separation decreased significantly (p<0.05) compared to the titanium implants in diabetic rats. Histological examination revealed a greater amount of new bone tissue forming around the MSC-implant complexes and a higher bone implant contact (BIC) rate than the titanium implants. These findings demonstrate that MSC-implant complexes possess osteogenic abilities and can be used in diabetic rats to improve the BIC rate. Thus, MSC-implant complexes provide a novel tissue engineering approach that promotes osseous healing and may potentially be useful in the treatment of diabetic patients.