Objective The goal of this study was to develop a decellularized tendon scaffold(DTS)and repopulate it with adipose-derived stem cells(ADSCs)assisted by low air pressure(LP).Methods The porcine superficial flexor tend...Objective The goal of this study was to develop a decellularized tendon scaffold(DTS)and repopulate it with adipose-derived stem cells(ADSCs)assisted by low air pressure(LP).Methods The porcine superficial flexor tendons were processed into the DTSs using a combination of physical,chemical,and enzymatic treatments.The effectiveness of decellularization was verified by histological analysis and DNA quantification.The properties of the DTSs were evaluated by quantitative analysis of biochemical characterization,porosimetry,in vitro biocompatibility assessment,and biomechanical testing.Subsequently,the ADSCs-DTS complexes were constructed via cell injection assisted by LP or under atmospheric pressure.The differences in cell distribution,biomechanical properties,and the total DNA content were compared by histological analysis,biomechanical testing,and DNA quantification,respectively.Results Histological analysis confirmed that no cells or condensed nuclear materials were retained within the DTSs with widened interfibrillar space.The decellularization treatment resulted in a significant decrease in the content of DNA and glycosaminoglycans,and a significant increase in the porosity.The DTSs were cytocompatible in vitro and did not show reduced collagen content and inferior biomechanical properties compared with the fresh-frozen tendons.The assistance of LP promoted the broader distribution of cells into the adjacent interfibrillar space and cell proliferation in DTSs.The biomechanical properties of the scaffolds were not significantly affected by the recellularization treatments.Conclusion A novel LP-assisted approach for the construction of cells-DTS complex was established,which could be a methodological foundation for further bioreactor and in vitro studies.展开更多
Anaplastic thyroid carcinoma(ATC)is a rare but extremely lethal malignancy.However,little is known about the pathogenesis of ATC.Given its high mortality,it is critical to improve our understanding of ATC pathogenesis...Anaplastic thyroid carcinoma(ATC)is a rare but extremely lethal malignancy.However,little is known about the pathogenesis of ATC.Given its high mortality,it is critical to improve our understanding of ATC pathogenesis and to find new diagnostic biomarkers.In the present study,two gene microarray profiles(GSE53072 and GSE65144),which included 17 ATC and 17 adjacent non-tumorous tissues,were obtained.Bioinformatic analyses were then performed.Immunohistochemistry(IHC)and receiver operating characteristic(ROC)curves were then used to detect transmembrane protein 158(TMEM158)expression and to assess diagnostic sensitivity.A total of 372 differentially expressed genes(DEGs)were identified.Through protein-protein interaction(PPI)analysis,we identified a significant module with 37 upregulated genes.Most of the genes in this module were related to cell-cycle processes.After co-expression analysis,132 hub genes were selected for further study.Nine genes were identified as both DEGs and genes of interest in the weighted gene co-expression network analysis(WGCNA).IHC and ROC curves confirmed that TMEM158 was overexpressed in ATC tissue as compared with other types of thyroid cancer and normal tissue samples.We identified 8 KEGG pathways that were associated with high expression of TMEM158,including aminoacyl-tRNA biosynthesis and DNA replication.Our results suggest that TMEM158 may be a potential oncogene and serve as a diagnostic indicator for ATC.展开更多
基金the National Natural Science Foundation of China(No.81672166).
文摘Objective The goal of this study was to develop a decellularized tendon scaffold(DTS)and repopulate it with adipose-derived stem cells(ADSCs)assisted by low air pressure(LP).Methods The porcine superficial flexor tendons were processed into the DTSs using a combination of physical,chemical,and enzymatic treatments.The effectiveness of decellularization was verified by histological analysis and DNA quantification.The properties of the DTSs were evaluated by quantitative analysis of biochemical characterization,porosimetry,in vitro biocompatibility assessment,and biomechanical testing.Subsequently,the ADSCs-DTS complexes were constructed via cell injection assisted by LP or under atmospheric pressure.The differences in cell distribution,biomechanical properties,and the total DNA content were compared by histological analysis,biomechanical testing,and DNA quantification,respectively.Results Histological analysis confirmed that no cells or condensed nuclear materials were retained within the DTSs with widened interfibrillar space.The decellularization treatment resulted in a significant decrease in the content of DNA and glycosaminoglycans,and a significant increase in the porosity.The DTSs were cytocompatible in vitro and did not show reduced collagen content and inferior biomechanical properties compared with the fresh-frozen tendons.The assistance of LP promoted the broader distribution of cells into the adjacent interfibrillar space and cell proliferation in DTSs.The biomechanical properties of the scaffolds were not significantly affected by the recellularization treatments.Conclusion A novel LP-assisted approach for the construction of cells-DTS complex was established,which could be a methodological foundation for further bioreactor and in vitro studies.
基金This study was supported by grants from Tongji Medical College,Huazhong University of Science and Technology(CN)(No.5001540018)Young Scientists Fund(No.81802676).
文摘Anaplastic thyroid carcinoma(ATC)is a rare but extremely lethal malignancy.However,little is known about the pathogenesis of ATC.Given its high mortality,it is critical to improve our understanding of ATC pathogenesis and to find new diagnostic biomarkers.In the present study,two gene microarray profiles(GSE53072 and GSE65144),which included 17 ATC and 17 adjacent non-tumorous tissues,were obtained.Bioinformatic analyses were then performed.Immunohistochemistry(IHC)and receiver operating characteristic(ROC)curves were then used to detect transmembrane protein 158(TMEM158)expression and to assess diagnostic sensitivity.A total of 372 differentially expressed genes(DEGs)were identified.Through protein-protein interaction(PPI)analysis,we identified a significant module with 37 upregulated genes.Most of the genes in this module were related to cell-cycle processes.After co-expression analysis,132 hub genes were selected for further study.Nine genes were identified as both DEGs and genes of interest in the weighted gene co-expression network analysis(WGCNA).IHC and ROC curves confirmed that TMEM158 was overexpressed in ATC tissue as compared with other types of thyroid cancer and normal tissue samples.We identified 8 KEGG pathways that were associated with high expression of TMEM158,including aminoacyl-tRNA biosynthesis and DNA replication.Our results suggest that TMEM158 may be a potential oncogene and serve as a diagnostic indicator for ATC.