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The triggering receptor expressed on myeloid cells 2 (TREM2) is an immune receptor that affects cellular phenotypes by modulating phagocytosis and metabolism, promoting cell survival, and counteracting inflammation. Its role in renal injury, in particular, unilateral ureteral obstruction (UUO) or ischemia-reperfusion injury (IRI)-induced renal injury remains unclear. In our study, WT and Trem2-/- mice were employed to evaluate the role of TREM2 in renal macrophage infiltration and tissue injury after UUO. Bone marrow-derived macrophages (BMDM) from both mouse genotypes were cultured and polarized for in vitro experiments. Next, the effects of TREM2 on renal injury and macrophage polarization in IRI mice were also explored. We found that TREM2 expression was upregulated in the obstructed kidneys. TREM2 deficiency exacerbated renal inflammation and fibrosis 3 and 7 days after UUO, in association with reduced macrophage infiltration. Trem2-/- BMDM exhibited increased apoptosis and poorer survival compared with WT BMDM. Meanwhile, TREM2 deficiency augmented M1 and M2 polarization after UUO. Consistent with the in vivo observations, TREM2 deficiency led to increased polarization of BMDM towards the M1 proinflammatory phenotype. Mechanistically, TREM2 deficiency promoted M1 and M2 polarization via the JAK-STAT pathway in the presence of TGF-ß1, thereby affecting cell survival by regulating mTOR signaling. Furthermore, cyclocreatine supplementation alleviated cell death caused by TREM2 deficiency. Additionally, we found that TREM2 deficiency promoted renal injury, fibrosis, and macrophage polarization in IRI mice. The current data suggest that TREM2 deficiency aggravates renal injury by promoting macrophage apoptosis and polarization via the JAK-STAT pathway. These findings have implications for the role of TREM2 in the regulation of renal injury that justify further evaluation.
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Apoptose , Macrófagos , Glicoproteínas de Membrana , Camundongos Endogâmicos C57BL , Receptores Imunológicos , Fatores de Transcrição STAT , Transdução de Sinais , Animais , Macrófagos/metabolismo , Receptores Imunológicos/metabolismo , Receptores Imunológicos/deficiência , Receptores Imunológicos/genética , Glicoproteínas de Membrana/metabolismo , Glicoproteínas de Membrana/deficiência , Glicoproteínas de Membrana/genética , Camundongos , Fatores de Transcrição STAT/metabolismo , Janus Quinases/metabolismo , Rim/patologia , Rim/metabolismo , Camundongos Knockout , Masculino , Fibrose , Traumatismo por Reperfusão/patologia , Traumatismo por Reperfusão/metabolismo , Traumatismo por Reperfusão/genética , Obstrução Ureteral/patologia , Obstrução Ureteral/metabolismo , Obstrução Ureteral/complicações , Polaridade Celular , Serina-Treonina Quinases TOR/metabolismo , Injúria Renal Aguda/metabolismo , Injúria Renal Aguda/patologia , Injúria Renal Aguda/genéticaRESUMO
OBJECTIVE: Patients with laryngotracheal stenosis (LTS) often have dysphagia after laryngotracheal reconstruction with T-tube insertion, which affects the quality of life. The purpose of this study is to observe the effect of swallowing rehabilitation therapy on the improvement of quality of life in patients of otolaryngology-head and neck surgery with dysphagia undergoing T-tube implantation treatment through longitudinal study. METHODS: Thirty-eight patients with LTS who experienced dysphagia after laryngotracheal reconstruction and T-tube implantation were recruited. All patients received swallowing rehabilitation therapy. The assessment of swallowing function was performed using the 10-item Eating Assessment Tool (EAT-10), the 30 mL water swallow test (WST), and flexible endoscopic evaluation of swallow (FEES). RESULTS: After swallowing rehabilitation therapy, timing of swallowing, grade of dysphagia, performance on FEES and 30 mL WST, and EAT-10 score all improved. Thirty-eight patients successfully transitioned to oral feeding and were able to remove their nasogastric tubes without experiencing any complications, including aspiration pneumonia. CONCLUSION: For patients with LTS who experienced dysphagia after laryngotracheal reconstruction and T-tube implantation, swallowing rehabilitation therapy could improve swallowing function of the patients, so as to reduce the potential harm caused by the pain and complications of surgery experienced by patients.
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BACKGROUND: T cell dysfunction, which includes exhaustion, anergy, and senescence, is a distinct T cell differentiation state that occurs after antigen exposure. Although T cell dysfunction has been a cornerstone of cancer immunotherapy, its potential in transplant research, while not yet as extensively explored, is attracting growing interest. Interferon regulatory factor 4 (IRF4) has been shown to play a pivotal role in inducing T cell dysfunction. METHODS: A novel ultra-low-dose combination of Trametinib and Rapamycin, targeting IRF4 inhibition, was employed to investigate T cell proliferation, apoptosis, cytokine secretion, expression of T-cell dysfunction-associated molecules, effects of MAPK and mammalian target of Rapamycin (mTOR) signaling pathways, and allograft survival in both in vitro and BALB/c to C57BL/6 mouse cardiac transplantation models. RESULTS: In vitro, blockade of IRF4 in T cells effectively inhibited T cell proliferation, increased apoptosis, and significantly upregulated the expression of programmed cell death protein 1 (PD-1), Helios, CD160, and cytotoxic T lymphocyte-associated antigen (CTLA-4), markers of T cell dysfunction. Furthermore, it suppressed the secretion of pro-inflammatory cytokines interferon (IFN)-γ and interleukin (IL)-17. Combining ultra-low-dose Trametinib (0.1 mg·kg-1·day-1) and Rapamycin (0.1 mg·kg-1·day-1) demonstrably extended graft survival, with 4 out of 5 mice exceeding 100 days post-transplantation. Moreover, analysis of grafts at day 7 confirmed sustained IFN regulatory factor 4 (IRF4) inhibition, enhanced PD-1 expression, and suppressed IFN-γ secretion, reinforcing the in vivo efficacy of this IRF4-targeting approach. The combination of Trametinib and Rapamycin synergistically inhibited the MAPK and mTOR signaling network, leading to a more pronounced suppression of IRF4 expression. CONCLUSIONS: Targeting IRF4, a key regulator of T cell dysfunction, presents a promising avenue for inducing transplant immune tolerance. In this study, we demonstrate that a novel ultra-low-dose combination of Trametinib and Rapamycin synergistically suppresses the MAPK and mTOR signaling network, leading to profound IRF4 inhibition, promoting allograft acceptance, and offering a potential new therapeutic strategy for improved transplant outcomes. However, further research is necessary to elucidate the underlying pharmacological mechanisms and facilitate translation to clinical practice.
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OBJECTIVE: Tracheal transplantation could be a better option for patients with long segmental laryngotracheal stenosis or defects, but the need for immunosuppressants limits its widespread use due to the antigenicity of the tracheal epithelium. Chemically treated or cryopreserved nonviable tracheal allografts have no immunogenicity but lead to necrosis and stenosis in long-term outcomes. The present report describes the 5-year outcomes of de-epithelialized viable tracheal allotransplantation without immunosuppressants in a patient with severe laryngotracheal stenosis. METHODS: The recipient was a 47-year-old female with relapsing polychondritis affecting the larynx and cervical trachea and producing a 5 cm long stenosis that could not be repaired using resection and anastomosis. A tracheal allograft was obtained from a 45-year-old male donor and treated with a combination of 3% sodium dodecyl sulfate (SDS) and organ preservation solution for 138 hours. The allograft was revascularized by heterotopical implantation in the infrahyoid muscles of the recipient for 3 months and then transplantation to the laryngotracheal defect with a split-thickness skin graft sutured to the lumen and a silicon T-tube. No immunosuppressants were used postoperatively. RESULTS: The allograft was de-epithelialized, and most of the cartilage rings remained viable after the treatment. The allograft was revascularized, viable, and mechanically stable after 3 months of heterotopic implantation. No apparent signs of rejection or destruction were observed. The T-tube was removed, and the internal lining of the allograft was repopulated 4 months after orthotopic transplantation, despite the skin graft necrotizing at 2 weeks. Endoscopy and computed tomography showed a patent airway 5 years after orthotopic transplantation. The patient was able to resume her usual quality of life. CONCLUSION: The present study demonstrates that transplantation of the de-epithelialized viable tracheal allograft without immunosuppressants is safe and promising for patients with long laryngotracheal stenosis or defects, especially for those with malignant tumor resections.
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Traqueia , Estenose Traqueal , Humanos , Masculino , Feminino , Pessoa de Meia-Idade , Traqueia/transplante , Imunossupressores/uso terapêutico , Constrição Patológica/patologia , Seguimentos , Qualidade de Vida , Estenose Traqueal/cirurgiaRESUMO
BACKGROUND: Homoharringtonine (HHT) is an effective anti-inflammatory, anti-viral, and anti-tumor protein synthesis inhibitor that has been applied clinically. Here, we explored the therapeutic effects of HHT in a mouse heart transplant model. METHODS: Healthy C57BL/6 mice were used to observe the toxicity of HHT in the liver, kidney, and hematology. A mouse heart transplantation model was constructed, and the potential mechanism of HHT prolonging allograft survival was evaluated using Kaplan-Meier analysis, immunostaining, and bulk RNA sequencing analysis. The HHT-T cell crosstalk was modeled ex vivo to further verify the molecular mechanism of HHT-induced regulatory T cells (Tregs) differentiation. RESULTS: HHT inhibited the activation and proliferation of T cells and promoted their apoptosis ex vivo. Treatment of 0.5 mg/kg HHT for 10 days significantly prolonged the mean graft survival time of the allografts from 7 days to 48 days (P <0.001) without non-immune toxicity. The allografts had long-term survival after continuous HHT treatment for 28 days. HHT significantly reduced lymphocyte infiltration in the graft, and interferon-γ-secreting CD4+ and CD8+ T cells in the spleen (P <0.01). HHT significantly increased the number of peripheral Tregs (about 20%, P <0.001) and serum interleukin (IL)-10 levels. HHT downregulated the expression of T cell receptor (TCR) signaling pathway-related genes (CD4, H2-Eb1, TRAT1, and CD74) and upregulated the expression of IL-10 and transforming growth factor (TGF)-ß pathway-related genes and Treg signature genes (CTLA4, Foxp3, CD74, and ICOS). HHT increased CD4+ Foxp3+ cells and Foxp3 expression ex vivo, and it enhanced the inhibitory function of inducible Tregs. CONCLUSIONS: HHT promotes Treg cell differentiation and enhances Treg suppressive function by attenuating the TCR signaling pathway and upregulating the expression of Treg signature genes and IL-10 levels, thereby promoting mouse heart allograft acceptance. These findings may have therapeutic implications for organ transplant recipients, particularly those with viral infections and malignancies, which require a more suitable anti-rejection medication.
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Disulfide bond A oxidoreductase-like protein (DsbA-L) drives acute kidney injury (AKI) by directly upregulating the expression of voltage-dependent anion-selective channels in proximal tubular cells. However, the role of DsbA-L in immune cells remains unclear. In this study, we used an LPS-induced AKI mouse model to assess the hypothesis that DsbA-L deletion attenuates LPS-induced AKI and explore the potential mechanism of DsbA-L action. After 24 hours of LPS exposure, the DsbA-L knockout group exhibited lower serum creatinine levels compared to the WT group. Furthermore, peripheral levels of the inflammatory cytokine IL-6 were decreased. Transcriptomic data analysis revealed a significant down-regulation in the IL-17 and tumor necrosis factor pathways in DsbA-L knockout mice following LPS induction. Metabolomic analysis suggested that arginine metabolism was significantly different between the WT and DsbA-L knockout groups after LPS treatment. Notably, the M1 polarization of macrophages in the kidneys of DsbA-L knockout AKI mice was significantly reduced. Expression of the transcription factors NF-κB and AP-1 was downregulated after DsbA-L knockout. Our results suggest that DsbA-L regulates LPS-mediated oxidative stress, promotes M1 polarization of macrophages, and induces expression of inflammatory factors via the NF-κB/AP-1 pathway.
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Injúria Renal Aguda , NF-kappa B , Animais , Camundongos , Injúria Renal Aguda/induzido quimicamente , Injúria Renal Aguda/genética , Rim/patologia , Lipopolissacarídeos/farmacologia , Macrófagos , NF-kappa B/metabolismo , Fator de Transcrição AP-1RESUMO
The calcineurin inhibitor-FK506-is a first-line immunosuppressant that regulates T cell secretion of IL-2 and other cytokines. However, the mechanism of its protective effect on target cells and its role on tumour recurrence and interaction with anti-tumour immune checkpoint inhibitors, such as PD-L1 blocking, are still unclear. Here, in a murine heart transplantation model, we observed the upregulation of programmed death-ligand 1 (PD-L1) expression by FK506 in both dendritic cells (DCs) and allografts. Blocking PD-L1 during FK506 treatment increased IFN-γ and TNF-α expression, enhanced CD4+ and CD8+ T cell proliferation, and suppressed Treg differentiation. Moreover, PD-L1 decreased T cell infiltration and induced T cell apoptosis in both the spleen and graft. PD-L1 was not only required in FK506-mediated immunosuppression but also upregulated by FK506. Treatment with SAFit2, a FKBP51 selective inhibitor, reduced the expression of PD-L1 on DCs and the grafts and interfered with the immunosuppressive effect of FK506, suggesting that the mechanism depends on FK506-binding protein (FKBP) 51 expression. Overall, our results add new insights into the role of FK506, not only on T cell cytokine secretion but also on co-inhibitory molecular regulation and target cell immune privilege.
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Transplante de Coração , Tacrolimo , Animais , Antígeno B7-H1/metabolismo , Camundongos , Receptor de Morte Celular Programada 1/metabolismo , Tacrolimo/farmacologia , Proteínas de Ligação a Tacrolimo/metabolismo , Regulação para CimaRESUMO
Perfusion-decellularization was an interesting technique to generate a natural extracellular matrix (ECM) with the complete three-dimensional anatomical structure and vascular system. In this study, the esophageal ECM (E-ECM) scaffold was successfully constructed by perfusion-decellularized technique through the vascular system for the first time. And the physicochemical and biological properties of the E-ECM scaffolds were evaluated. The bone marrow mesenchymal stem cells (BMSCs) were induced to differentiate into myocytesin vitro. E-ECM scaffolds reseeded with myocytes were implanted into the greater omenta to obtain recellular esophageal ECM (RE-ECM), a tissue-engineered esophagus. The results showed that the cells of the esophagi were completely and uniformly removed after perfusion. E-ECM scaffolds retained the original four-layer organizational structure and vascular system with excellent biocompatibility. And the E-ECM scaffolds had no significant difference in mechanical properties comparing with fresh esophagi,p> 0.05. Immunocytochemistry showed positive expression ofα-sarcomeric actin, suggesting that BMSCs had successfully differentiated into myocytes. Most importantly, we found that in the RE-ECM muscularis, the myocytes regenerated linearly and continuously and migrated to the deep, and the tissue vascularization was obvious. The cell survival rates at 1 week and 2 weeks were 98.5 ± 3.0% and 96.4 ± 4.6%, respectively. It was demonstrated that myocytes maintained the ability for proliferation and differentiation for at least 2 weeks, and the cell activity was satisfactory in the RE-ECM. It follows that the tissue-engineered esophagus based on perfusion-decellularized technique and mesenchymal stem cells has great potential in esophageal repair. It is proposed as a promising alternative for reconstruction of esophageal defects in the future.
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Matriz Extracelular Descelularizada/química , Esôfago , Células-Tronco Mesenquimais/citologia , Engenharia Tecidual/métodos , Animais , Células Cultivadas , Esôfago/química , Esôfago/citologia , Esôfago/metabolismo , Masculino , Perfusão , CoelhosRESUMO
BACKGROUND: As an excellent xenotransplant, the pig trachea can be decellularized and cryopreserved to reduce its immunogenicity. However, few reports are found on the changes of its mechanical properties after cryopreservation and decellularization. OBJECTIVE: To evaluate the structure and biomechanical properties in pig tracheal scaffolds resulting from decellularized and cryopreserved. MATERIAL AND METHODS: Twenty-five pig tracheal segments were separated into five groups: untreated (group A), only decellularized (group B), only cryopreserved (group C), decellularized after cryopreserved (group D) and cryopreserved after decellularized (group E). Tracheal segments were subjected to uniaxial tension or compression using a universal testing machine to determine structural biomechanical changes. RESULTS: It showed that there was no statistically significant difference in the tensile strength of the trachea in each group. The compressive strength of group B, C and D were same as the group A (P > 0.05), while the group E was lower than that of the group A (P < 0.05).Conclusions and significance: The histological examination of the decellularization after cryopreservation shows that the removal of epithelial cells and submucosal cells is more thorough, and the biomechanical structure of the trachea is better preserved. This proved to be a new method to prepare xenotransplantation of trachea graft.
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Criopreservação/métodos , Alicerces Teciduais , Traqueia , Animais , Fenômenos Biomecânicos , Força Compressiva , DNA/análise , DNA/química , Células Epiteliais , Masculino , Fenazinas , Coloração e Rotulagem/métodos , Suínos , Resistência à Tração , Alicerces Teciduais/química , Traqueia/citologiaRESUMO
OBJECTIVES: Reconstruction of long segmental tracheal defects is difficult because no ideal tracheal substitutes are currently available. Tracheal allotransplantation maintains cartilage and epithelium viability but requires immunosuppression because of epithelial immunogenicity. We aimed to obtain an epithelium-decellularized allograft that maintains cartilage viability and to evaluate long-term outcomes of such allografts implanted on dog backs without immunosuppressants. METHODS: Twenty-five tracheas harvested from mongrel dogs were used to explore the period of epithelium decellularization by combined use of 1% sodium dodecyl sulfate and an organ preservation solution and to assess the chondrocyte viability and immunogenicity of the tracheas after decellularization. Sixteen epithelium-decellularized tracheal allografts and 10 fresh tracheal segments (6 cm long) were implanted in 26 beagles for durations of 10 days and 1, 3, 6, and 12 months. Macroscopic and microscopic examinations were used to evaluate the morphology, viability, and immune rejection of the allografts. Safranin-O staining was used to detect glycosaminoglycans. RESULTS: The epithelium disappeared after 24 hours of decellularization. At 72 hours, almost no nuclei remained in the mucosa, while the mean survival rate of chondrocytes was 88.1%. Histological analysis demonstrated that the allograft retained intact tracheal rings and viable cartilage after heterotopic implantation for 1 year, with no immunological rejection. There were no significant differences in the glycosaminoglycan contents among the implanted epithelium-decellularized allografts. CONCLUSIONS: Epithelium-decellularized tracheal allografts with chondrocyte viability can be achieved by combined use of a detergent and organ preservation solution, which showed satisfactory cartilage viability and structural integrity after long-term heterotopic transplantation. Further studies on orthotopic transplantation are needed to assess the feasibility of allografts in reconstructing long segmental tracheal defects.
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Cartilagem , Sobrevivência Celular , Condrócitos/fisiologia , Rejeição de Enxerto/prevenção & controle , Procedimentos de Cirurgia Plástica/métodos , Traqueia , Transplante Homólogo/métodos , Animais , Cartilagem/imunologia , Cartilagem/transplante , Modelos Animais de Doenças , Cães , Preservação de Órgãos/métodos , Traqueia/imunologia , Traqueia/transplante , Imunologia de Transplantes , Resultado do TratamentoRESUMO
Objective:The aim of this study is to o explore the diagnosis, treatment and prognosis of idiopathic subglottic stenosis ï¼ISSï¼ Method:The clinical data of 15 patients with idiopathic subglottic stenosis treated in our department were analyzed retrospectively. The degree of stenosis was classified by the Cotton Airway grading system of Myer, with 8 cases of gradeâ ¡, 4 cases of grade â ¢ and 3 cases of grade â £. Result:The time of follow-up of HTSS was 0.5-10 years. All 15 patients were successfully extubated without asphyxia, decannulation and wound nonunion. Conclusion:For patients with idiopathic subglottic stenosis in the non-progressive stage, active surgical treatment strategy should be adopted and treated individually. The prognosis is satisfactory.
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Laringoestenose/diagnóstico , Laringoestenose/cirurgia , Procedimentos de Cirurgia Plástica , Constrição Patológica , Humanos , Prognóstico , Estudos Retrospectivos , Resultado do TratamentoRESUMO
OBJECTIVE: To systematically study various surgical approaches for treating complex hypopharyngeal and/or laryngotracheal stenoses at a variety of sites and levels. PATIENTS AND METHODS: We retrospectively analyzed the treatment of 17 patients with severe and complex hypopharyngeal and/or laryngotracheal stenosis at various sites and levels of severity. All of the 17 patients initially had a tracheostomy. Thirteen had failed the previous laser lysis and/or dilation treatment. Given the high severity and complexity of stenosis, all of these patients were treated by open surgical reconstruction techniques using repairing grafts (flaps), followed by stenting. RESULTS: Thirteen of 17 patients had successful decannulation 1-8 months post-operation and had stable airway and adequate vocal and swallow function. Two patients with complex hypopharyngeal and esophageal stenosis had unsuccessful decannulation. Follow-up was lost in 1 patient with complex hypopharyngeal and esophageal stenosis and 1 patient with original hypopharyngeal stenosis and recurrent thoracotracheal stenosis. CONCLUSION: Despite the failure by the regular treatments using laser lysis and/or dilation therapy, severe and complex hypopharyngeal and/or laryngotracheal stenosis may be successfully treated by variable open surgical reconstruction techniques using different grafts (flaps) depending on the site and severity of the stenosis.
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Laringoestenose/cirurgia , Terapia a Laser/métodos , Procedimentos de Cirurgia Plástica/métodos , Complicações Pós-Operatórias/cirurgia , Retalhos Cirúrgicos , Estenose Traqueal/cirurgia , Adolescente , Adulto , Criança , Pré-Escolar , Feminino , Seguimentos , Humanos , Laringoplastia/métodos , Laringoestenose/etiologia , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/etiologia , Estudos Retrospectivos , Fatores de Tempo , Estenose Traqueal/etiologia , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Increasing evidence indicates that dysregulation of microRNAs is involved in tumor progression and development. The aim of this study was to investigate the expression of microRNA-24 (miR-24) and its function in laryngeal squamous cell carcinoma (LSCC). METHODS: Quantitative RT-PCR (qRT-PCR) was used to detect miR-24 expression in LSCC cell lines and tissue samples. MTT, colony formation, and flow cytometry was performed to analyze the effects of miR-24 expression on growth, apoptosis, and radiosensitivity of LSCC cells. Dual-luciferase reporter assays were performed to examine regulation of putative miR-24 targets. Expression of X-linked inhibitor of apoptosis protein (XIAP) mRNA and protein, cleaved or total caspase-3, and cleaved or total PARP protein were detected by qRT-PCR and western blotting assays, respectively. RESULTS: miR-24 expression levels in LSCC cell lines or tissue were significantly lower than in a normal human keratinocyte cell line or adjacent normal tissues. Functional analyses indicated that re-expression of miR-24 inhibits growth, reduces colony formation, and enhances apoptosis in LSCC cells. In addition, miR-24 upregulation increases LSCC sensitivity to irradiation by enhancing irradiation-induced apoptosis, and luciferase activity indicated that miR-24 binds to the 3'-untranslated region (3'-UTR) of XIAP mRNA. Upregulation of miR-24 inhibits XIAP protein expression in LSCC cells, and silencing of XIAP mimics the effects of miR-24 upregulation on LSCC cells. In addition, XIAP mRNA expression significantly increases in LSCC tissues and is inversely correlated with miR-24 expression. CONCLUSIONS: Our data suggest that miR-24 inhibits growth, increases apoptosis, and enhances radiosensitivity in LSCC cells by targeting XIAP. Therefore, miR-24 may be a potential molecular target for the treatment of human LSCC.
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OBJECTIVE: To explore the survivorship and character of decellularized laryngeal scaffold in pectoralis major muscle flap in canine. METHODS: Eighteen donor larynx in experimental group were decellularized by perfusing sodium dodecyl sulphate. Three of them were used to detect the character of histology. The other fifteen ones were embedded in right pectoralis major muscle flap of acceptor canine. Donor larynx in control group were not perfused. Other experimental procedure was the same as experimental group. The specimens were harvested at two weeks, one month and two months after operation, respectively. Macroscopic view, histological examination and trypan blue staining were performed in the experimental group and control group. RESULTS: The size of the specimens decreased remarkably into disappearance in control group, there was statistical significance between the experimental group and the control group (which used least significant difference t test P < 0.05). There was only little neutrophils and lymphocytes infiltrating around the laryngeal scaffold at 2 weeks in the experimental group. One month after operation, loose connective tissue begin to form around the laryngeal scaffold. After two months of transplantation, the connective tissue became thicker and the number of blood vessels increased than before. There was a large number of lymphocytes and neutrophil infiltration around the laryngeal specimens in the control group at 2nd week. The perichondrium in the control group was damaged at one month post operation. The cartilage cells could not be detected two months after surgery. The survival rate of cartilage cell between experimental group (86.8% ± 3.2%) and the control group (88.6% ± 3.1%) did not show statistical significance before implantation (χ(2) = 0.19, P > 0.05). The survival rate of cartilage cell decreased insignificantly in experimental group while the survival rate declined obviously in the control group at two weeks and one month after operation, the difference had statistical significance (χ(2) were respectively 5.52 and 20.55, P were respectively < 0.05 and < 0.01), the survival rate of cartilage cell in experimental group was (65.8% ± 2.6%) at two months after operation, while the cartilage cell all disappeared in control group. CONCLUSIONS: Perfused decellularation technique can construct a low immunogenicity laryngeal cartilage scaffold which can survive in the chest muscle package and establish a good blood supplement. The decellularized laryngeal scaffold could be used as a biological scaffold for whole laryngeal reconstruction.
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Condrócitos/citologia , Laringe Artificial , Alicerces Teciduais , Animais , Técnicas de Cultura de Células , Cães , Engenharia Tecidual/métodosRESUMO
Previous studies have shown that miR-203 acts as a tumor-suppressive microRNA in various cancers, but its roles in laryngeal carcinoma are still contradicted. Here, we found that miR-203 inhibited the growth of laryngeal cancer cells and survivin was a direct target of miR-203. Moreover, silencing of survivin recapitulated the effect of miR-203 on cell cycle progression, whereas overexpression of survivin reversed this effect. Additionally, qRT-PCR showed the reciprocal relationship between miR-203 and survivin in laryngeal cancer tissues. These findings indicate that miR-203 inhibits the proliferation of laryngeal carcinoma cells by directly targeting survivin, suggesting its application in anti-cancer therapeutics.
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Carcinoma/genética , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Proteínas Inibidoras de Apoptose/metabolismo , Neoplasias Laríngeas/genética , MicroRNAs/metabolismo , Carcinoma/patologia , Linhagem Celular Tumoral , Humanos , Proteínas Inibidoras de Apoptose/genética , Neoplasias Laríngeas/patologia , MicroRNAs/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , SurvivinaRESUMO
OBJECTIVE: The current methods of management of tracheal stenosis have disadvantages and are controversial, therefore ideal experimental animal models for the further studies are required. The aim of this study was to establish a new model of canine tracheal stenosis by radiofrequency cauterization. METHODS: The tracheal cartilage ring was injured by radiofrequency at the level of the sixth tracheal ring in 12 mongrel dogs. A fibrolaryngoscope was used to guide the procedure. The animals were observed after operation and examined after euthanasia. Endoscopic and histological examinations were undertaken to evaluate the progress of stenosis. The degree of stenosis was calculated using the formula: degree of stenosis=(initial lumen area-final lumen area)/initial lumen area × 100%. RESULTS: Tracheal stenosis had developed in all dogs by the 21st day post operation. Costal retraction was observed in all dogs after the 18th day post operation. At the end of the study, gross and endoscopic examinations showed that stenosis had been induced to a satisfactory degree and without any complications. The median of the degree of stenosis was 92%, with a range of 84-94%. Histological examination showed that cartilage was damaged and that granulation tissue and collagen fibres had formed. CONCLUSIONS: The model of canine tracheal stenosis induced by radiofrequency cauterization is a relatively simple, reliable, and reproducible animal model. This model may be useful in the development of new methods of treatment for tracheal stenosis.