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In this work, PDA@BN-TiO2 nanohybrid particles were incorporated chemically into a zinc-phosphating solution to form a robust, low-temperature phosphate-silane coating on Q235 steel specimens. The morphology and surface modification of the coating was characterized by X-Ray Diffraction (XRD), X-ray Spectroscopy (XPS), Fourier-transform infrared spectroscopy (FT-IR), and Scanning electron microscopy (SEM). Results demonstrate that the incorporation of PDA@BN-TiO2 nanohybrids produced a higher number of nucleation sites and reduced grain size with a denser, more robust, and more corrosion-resistant phosphate coating compared to pure coating. The coating weight results showed that the PBT-0.3 sample achieved the densest and most uniform coating (38.2 g/m2). The potentiodynamic polarization results showed that the PDA@BN-TiO2 nanohybrid particles increased phosphate-silane films' homogeneity and anti-corrosive capabilities. The 0.3 g/L sample exhibits the best performance with an electric current density of 1.95 × 10-5 A/cm2, an order of magnitude lower than that of the pure coatings. Electrochemical impedance spectroscopy revealed that PDA@BN-TiO2 nanohybrids provided the greatest corrosion resistance compared to pure coatings. The corrosion time for copper sulfate in samples containing PDA@BN/TiO2 prolonged to 285 s, a significantly higher amount of time than the corrosion time found in pure samples.
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BACKGROUND: Tympanic membrane perforation (TMP) is a common disease in otology, and few acellular techniques have been reported for repairing this condition. Decellularized extracellular matrix (ECM) scaffolds have been used in organ reconstruction. OBJECTIVE: This study on tissue engineering aimed to develop a tympanic membrane (TM) scaffold prepared using detergent immersion and bone marrow mesenchymal stem cells (BMSCs) as repair materials to reconstruct the TM. RESULTS: General structure was observed that the decellularized TM scaffold with BMSCs retained the original intact anatomical ECM structure, with no cell residue, as observed using scanning electron microscopy (SEM), and exhibited low immunogenicity. Therefore, we seeded the decellularized TM scaffold with BMSCs for recellularization. Histology and eosin staining, SEM and immunofluorescence in vivo showed that the recellularized TM patch had a natural ultrastructure and was suitable for the migration and proliferation of BMSCs. The auditory brainstem response (ABR) evaluated after recellularized TM patch repair was slightly higher than that of the normal TM, but the difference was not significant. CONCLUSION: The synthetic ECM scaffold provides temporary physical support for the three-dimensional growth of cells during the tissue developmental stage. The scaffold stimulates cells to secrete their own ECM required for tissue regeneration. The recellularized TM patch shows potential as a natural, ultrastructure biological material for TM reconstruction.
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Células-Tronco Mesenquimais , Perfuração da Membrana Timpânica , Humanos , Alicerces Teciduais/química , Matriz Extracelular/química , Perfuração da Membrana Timpânica/terapia , Membrana Timpânica , Engenharia Tecidual/métodos , Células da Medula ÓsseaRESUMO
Acute respiratory failure (ARF) with a high incidence among moderate-to-severe traumatic brain injury (M-STBI) patients plays a pivotal role in worsening neurological outcomes. Traumatic subarachnoid hemorrhage (tSAH) is highly prevalent in M-STBI, which is associated with significant adverse outcomes. In this retrospective cohort study, we aimed to explore the association between the severity of the tSAH and ARF in the M-STBI population. A total of 771 subjects were reviewed. Clinical and neuroimaging data of M-STBI patients were retrospectively collected, and ARF was ascertained retrospectively based on their electronic medical record. The degree of tSAH was classified according to Fisher's criteria, and the grade of tSAH was dichotomized to a low Fisher grade (Fisher grade 1-2) and a high Fisher grade (Fisher grade 3-4). After exclusion procedures, the data of 695 M-STBI patients were analyzed. A total of 284 (30.8%) had a high Fisher grade on admission. The overall rate of ARF within 48 h upon admission was 34.4% (239/695); it was 29.5% (142/481) and 46.3% (99/214) for the low and high Fisher groups, respectively. In a full cohort, a high Fisher grade was associated with ARF after adjusting for age, gender, GCS, smoking history, comorbidities, multiple injuries, characteristics of TBI, and pulmonary factors (OR 1.78; 95% CI, 1.11-2.85, p = 0.016). This result remained robust in the comparisons after PSM (71/132, 42.8% vs. 53/132, 31.9%; OR, 1.59; 95% CI, 1.02-2.49, p = 0.042). A high Fisher SAH grade exposure on admission is associated with ARF in M-STBI patients.
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BACKGROUND: Hypopharynx reconstruction after hypopharyngectomy is still a great challenge. Perfusion decellularization is for extracellular matrix (ECM) scaffolding and had been used in organ reconstruction. Our study aimed to prepare an acellular, natural, three-dimensional biological hypopharynx with vascular pedicle scaffold as the substitute materials to reconstruct hypopharynx. RESULT: Scanning electron microscope and histology staining showed that the decellularized hypopharynx with vascular pedicle scaffold retained intact native anatomical ECM structure. Myoblasts were observed on the recellularized scaffolds with bone marrow mesenchymal stem cells induced by 5-azacytidine implanted in the rabbit greater omentum by immunohistochemical analysis. CONCLUSION: The decellularized hypopharynx with vascular pedicle scaffold prepared by detergent perfusion in our study has a potential to be an alternative material to pharynx reconstruction.
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Células-Tronco Mesenquimais , Alicerces Teciduais , Animais , Matriz Extracelular/química , Hipofaringe/cirurgia , Perfusão , Coelhos , Engenharia Tecidual/métodos , Alicerces Teciduais/químicaRESUMO
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
BACKGROUND: The etiology and distribution of community-acquired pneumonia (CAP) vary periodically and geographically. The clinical evaluation of CAP among patients with cancers remains unknown. PATIENTS AND METHODS: This retrospective hospital-based study on adult CAP was conducted in Tang Du Hospital, China, from September 2018 to August 2019. The demographic characteristics, clinical manifestations and laboratory data were extracted from medical records and compared between CAP patients with and without cancers. Univariable and multivariable logistic regression methods were used to explore risk factors associated with CAP patients with and without cancers. RESULTS: Data from 149 CAP patients with cancers and 268 CAP patients without cancers were analyzed. Patients without cancers were more likely to show fever, cough and yellow sputum, higher level of neutrophil count than the cancer patients. Klebsiella pneumoniae (K. pneumoniae 14.77% vs 9.33%, p = 0.093) and Streptococcus pneumoniae (S. pneumoniae 16.11% vs 11.57%, p = 0.189) were among the most commonly encountered pathogens in both the groups. Pseudomonas aeruginosa (P. pneumoniae 26.50% vs 11.41%, p < 0.001), Mycoplasma pneumoniae (M. pneumoniae 8.21% vs 1.34%, p = 0.003), and filamentous fungi (10.82% vs 4.7%, p = 0.033) were predominant in CAP patients without cancers. Haemophilus influenzae (H. influenzae 22.15% vs 14.18%, p = 0.038) and methicillin-resistant Staphylococci (MRS 23.49 vs 15.68, p = 0.049) were more prevalent for CAP cancer patients. Certain pathogens were increasing in a cold season. In patients without cancers, MRS, H. influenzae and P. aeruginosa were associated with central nervous system (CNS) disease, connective tissue disease, bronchiectasis, respectively. In addition, healthy adults were likely to be infected with M. pneumoniae showing fever. CONCLUSION: CAP patients with cancers had atypical clinical manifestations and showed no distinct increase in inflammatory markers. The predominant pathogens differed as well as similar between the CAP patients with and without cancers. Certain pathogens follow a seasonal pattern. CNS disease, connective tissue disease and bronchiectasis were associated with the predominant pathogens in patients without cancers.
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OBJECTIVE: To review the research progress of the tissue engineering technique in the esophageal defect repair and reconstruction. METHODS: The recently published clinical and experimental literature at home and abroad on the scaffold materials and the seeding cells used in the tissue engineered esophageal reconstruction was consulted and summarized. RESULTS: A large number of basic researches and clinical applications show that the effect of the tissue engineered esophagus is close to the autologous structure and function of the esophagus and it could be used for the repair of the esophageal defect. However, those techniques have a long distance from the clinical application and need an acknowledged rule of technology. CONCLUSION: Tissue engineering technique could provide an innovative theory for the esophageal defect reconstruction, but its clinical application need further research.
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Doenças do Esôfago/terapia , Esôfago/citologia , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Bioengenharia/métodos , Humanos , Medicina Regenerativa/tendênciasRESUMO
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
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.
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Cauterização/efeitos adversos , Modelos Animais de Doenças , Estenose Traqueal/patologia , Estenose Traqueal/cirurgia , Animais , Biópsia por Agulha , Broncoscopia , Cauterização/métodos , Cães , Feminino , Imuno-Histoquímica , Masculino , Sensibilidade e Especificidade , Índice de Gravidade de Doença , Estenose Traqueal/etiologiaRESUMO
OBJECTIVE: To evaluate the biomechanical characteristics of the decellularized laryngeal scaffold. METHODS: Ten Chinese adult dogs were randomly divided into two groups: perfusion group (n = 5) and control group (n = 5). The acellular larynx scaffold was obtained from dogs through cranial thyroid arteries perfusion with detergents. Comparative examinations were performed by the macroscopic view, histological view (hematoxylin and eosin stain, Alcian blue stain and Masson stain), scanning electron microscope (SEM) and biomechanical properties between perfusion group and control group. RESULTS: Macroscopic view showed that the decellularized laryngeal scaffold appeared pale asphyxia. HE stain indicated that there were little acellular traces of muscle and mucosa. Alcian blue stain, Masson stain and scanning electron microscope (SEM) suggested that there were no obvious changes about glycosaminoglycan and collagen. The compressive modulus of thyroid cartilage was (1.06 ± 0.07) MPa (x(-) ± s) in experimental groups and (1.15 ± 0.11) MPa in control group, showing no significant difference (t = 1.424, P > 0.05), neither in compressive modulus of annular cartilage (1.68 ± 0.11) MPa in experimental groups and (1.67 ± 0.09) MPa in control group (t = 0.185, P > 0.05). The tensile strength of thyroid cartilage between experimental (5.74 ± 0.88) MPa and control groups (6.18 ± 1.33) MPa did not have the statistical significance (t = 0.627, P > 0.05). CONCLUSION: These results indicate that perfusion method can construct a perfect biomechanical acellular larynx scaffold which could be a better selection for laryngeal reconstruction with tissue engineering method.
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Laringe Artificial , Stents , Engenharia Tecidual , Alicerces Teciduais , Animais , Fenômenos Biomecânicos , Cartilagem Articular , Técnicas de Cultura de Células , Células Cultivadas , Cães , Resistência à TraçãoRESUMO
CONCLUSION: Reseeding mesenchymal stem cells (MSCs) into the decellularized laryngeal muscle matrix for construction of a tissue-engineered larynx is feasible. This in vivo maturation in the omentum could be the first step before in situ implantation of the construct. This construct could facilitate a tissue-engineered method for laryngeal reconstruction. OBJECTIVES: The extracellular matrix (ECM) and MSCs have been widely used for organ reconstruction. Our study aimed to prepare a soft tissue decellularized laryngeal scaffold with intact laryngeal cartilages utilizing a perfusion-decellularized technique, reseeding cells on it, and then construct a recellularized larynx. METHODS: Perfusion-decellularized larynges were obtained from 20 rabbits by perfusion of the common carotid arteries with detergents. Twelve perfused larynges were observed by macroscopic visualization, histological examination, scanning electron microscopy (SEM), and cartilage viability. The remaining eight perfusion constructs were reseeded with induced MSCs aspirated from eight receptor rabbits. Composites were transferred into greater omentums of receptor rabbits after adherence for 1 day in vitro. Rabbit larynges were harvested after 4 weeks and 8 weeks, respectively. Macroscopic visualization, histological examination, and immunohistochemistry were performed. RESULTS: Larynges perfused by sodium dodecyl sulfate became transparent after 2 h of perfusion. Histology and SEM indicated that the perfusion method showed a better decullularized effect. Almost no intact cells or nuclei were found, while more pores and collagen fibers were retained in the decellularized matrix. The chondrocyte vitality assay indicated that chondrocyte vitality was high. Vascularization was clearly seen by 4 weeks and relatively integrated cartilage frameworks remained by 8 weeks. Histological and immunohistochemical examinations clearly showed muscle bundles and vessels.
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Matriz Extracelular , Laringe/citologia , Células-Tronco Mesenquimais/citologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Animais , Sobrevivência Celular/fisiologia , Condrócitos/citologia , Cartilagens Laríngeas/citologia , Músculos Laríngeos/citologia , Laringe/irrigação sanguínea , Masculino , Microscopia Eletrônica de Varredura , Neovascularização Fisiológica/fisiologia , Omento , CoelhosRESUMO
OBJECTIVE: To evaluate the immunogenicity of the decellularized laryngeal scaffold. METHODS: Twelve perfused, decellularized laryngeal scaffolds were obtained from rabbits through common carotid artery perfusion with detergents. The twelve decellularized laryngeal scaffolds and the twelve fresh larynxes were then implanted in para-laryngeal muscles of rabbits and harvested after two weeks, four weeks, twelve weeks and twenty-four weeks, respectively. Macroscopic view, histological examination and lymphocyte infiltration test were performed. RESULTS: The decellularized larynxes were implanted and preserved the laryngeal extracellular matrix and laryngeal architecture. The decellularized larynx did not show obvious immunological rejection after implanted into the para-laryngeal muscles of the recipient rabbits. The volume of implanted larynx became smaller but retained cartilage scaffold. The larynxes in the control group presented the serious immunological rejection and the majority tissues of the larynxes were disintegrated and substituted by the fibrous connective tissues after four weeks. The peripheral tissues were damaged and necrotic at different degrees. The quantity of the lymphocyte infiltration in the control group was higher than that in the experiment group and the result had the statistical significance (P < 0.01). CONCLUSIONS: Perfused, decellularized technique can construct a low immune laryngeal cartilage scaffold which could be a satisfactory material for laryngeal repair.
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Rejeição de Enxerto/imunologia , Laringe Artificial , Implantação de Prótese , Alicerces Teciduais , Animais , Cartilagem/citologia , Células Cultivadas , Feminino , Linfócitos/imunologia , Masculino , Coelhos , Engenharia Tecidual/métodosRESUMO
OBJECTIVE: To prepare a decellularized whole laryngeal scaffold by utilizing a perfusion-decellularized technique, reseed cells on it, and construct decellularized laryngeal muscles. METHODS: Perfusion decellularized larynxes were obtained by common carotid arterious perfusion with detergents. Then they were performed by macroscopic view, histological examination, scanning electron microscopy (SEM) and cartilage viability. Decellularized laryngeal scaffold were then reseeded with inducted mesenchymal stem cells (MSCs). Composites were transferred into greater omentums of rabbits after one day's adherence and harvested after eight weeks. Macroscopic view, histological examination and immunohistochemistry were performed. RESULTS: Perfusion larynxes became transparent after two hours. Histology and SEM indicated that perfusion method showed better decullularized effect. More vintages and collagen fibers but no intact cell or nuclei were retained in the decellularized matrix. Porosity measured by Image pro plus 6.0 was 80.4% +/- 3.2% (x +/- s). Chondrocyte vitality assay indicated chondrocyte vitality rate in the perfusion group was 86.9% +/- 1.5%. After eight weeks, vascularization formed and integrated cartilage frameworks still remained. Histological examination could clearly show the presence of muscle bundles and vessels. Immunohistochemical examination indicated that sarcomeric-alpha actin expressed positively in corresponding areas. CONCLUSIONS: It is feasible to reseed MSCs into the decellularized laryngeal muscle matrix for constructing tissue-engineered laryngeal muscles. This in vivo maturation into the omentum could be the first step before in situ implantation of the construct.