Background The body is unable to repair and regenerate large area bone defects. Moreover, the repair capacity of articular cartilage is very limited. There has long been a lack of effective treatments for osteochondral lesions. The engineered tissue with biphase synthetic for osteochondral repair has become one of the hot research fields over the past few years. In this study, an integrated biomanufacturing platform was constructed with bone marrow mesenchymal stem cells (BMSCs)/porous tantalum (pTa) associated with chondrocytes/collagen membranes (CM) to repair large osteochondral defects in load-bearing areas of goats. Methods Twenty-four goats with a large osteochondral defect in the femoral heads of the left hind legs were randomly divided into three groups: eight were treated with chondrocytes/CM-BMSCs/pTa, eight were treated with pure CM-pTa composite, and the other eight goats were untreated. The repair effect was assessed by X-ray, gross observation, and histomorphology for 16 weeks after the operation. In addition, the biocompatibility of chondrocytes/CM-BMSCs/pTa was observed by flow cytometry, CCK8, immunocytochemistry, and Q-PCR. The characteristics of the chondrocytes/CM-BMSCs/pTa were evaluated using both scanning electron microscopy and mechanical testing machine. Results The integrated repair material consists of pTa, injectable fibrin sealant, and CM promoted adhesion and growth of BMSCs and chondrocytes. pTa played an important role in promoting the differentiation of BMSCs into osteoblasts. Three-dimensional CM maintained the phenotype of chondrocytes successfully and expressed chondrogenic genes highly. The in vivo study showed that after 16 weeks from implantation, osteochondral defects in almost half of the femoral heads had been successfully repaired by BMSC-loaded pTa associated with biomimetic 3D collagen-based scaffold. Conclusions The chondrocytes/CM-BMSCs/pTa demonstrated significant therapeutic efficacy in goat models of large osteochondral defect. This provides a novel therapeutic strategy for large osteochondral lesions in load-bearing areas caused by severe injury, necrosis, infection, degeneration, and tumor resection with a high profile of safety, effectiveness, and simplicity.

BackgroundAs an ideal new graft material, porous tantalum (pTa) has excellent mechanical properties and corrosion resistance and has received increased attention in the biomedical field because of its excellent cytocompatibility and ability to induce bone formation. However, the molecular mechanism of its potential to promote osteogenesis remains unclear, and very few reports have been published on this topic.MethodsIn this study, we first produced porous Ti6Al4V (pTi6Al4V) and pTa with the same pore size by three-dimensional printing combined with chemical vapour deposition. The number of adhesions between pTa and pTi6Al4V and bone marrow mesenchymal stem cells (BMSCs) after 1 day of culture was detected by the live/dead cell staining method. The proliferation activity of the two groups was determined after culture for 1, 3, 5 and 7 days by the cell counting kit-8 method. In addition, the osteogenic activity, mRNA expression levels of osteogenic genes alkaline phosphatase (ALP), osterix (OSX), collagen-I (Col-I), osteonectin (OSN) and osteocalcin (OCN) and protein expression levels of the mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signalling pathway marker p-ERK of the two groups cultured for 7, 14 and 21 days were determined by the ALP activity assay, real-time quantitative polymerase chain reaction (Q-PCR) and Western blotting, respectively. Subsequently, the two groups were treated with the MAPK/ERK–specific inhibitor U0126, and then, the mRNA expression levels of osteogenic genes and protein expression levels of p-ERK in the cultures were determined by Q-PCR and Western blotting, respectively.ResultsThe live/dead cell staining and cell counting kit-8 assays showed that the adhesion and proliferation activities of BMSCs on pTa were significantly better than those on pTi6Al4V. In addition, the ALP activity assay and Q-PCR showed that pTa harboured osteogenic activity and that the osteogenic genes ALP, OSX, Col-I, OSN and OCN were highly expressed, and by Western blotting, the expression of p-ERK protein in the pTa group was also significantly higher than that in the pTi6Al4V group. Subsequently, using the MAPK/ERK–specific inhibitor U0126, Western blotting showed that the expression of p-ERK protein was significantly inhibited and that there was no difference between the two groups. Furthermore, Q-PCR showed that osteogenic gene expression and ALP expression levels were significantly increased in the pTa group, and there were no differences in the OSX, Col-I, OSN and OCN mRNA expression levels between the two groups.ConclusionOverall, our research found that compared with the widely used titanium alloy materials, our pTa can promote the adhesion and proliferation of BMSCs, and the molecular mechanism of pTa may occur via activation of the MAPK/ERK signalling pathway to regulate the high expression of OSX, Col I, OSN and OCN osteogenic genes and promote the osteogenic differentiation of BMSCs in vitro.The translational potential of this article: Our self-developed pTa mater...

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Differential expression of TRPV1 has been detected in many cancer types, including clear cell renal cell carcinoma (ccRCC). However, the clinical significance of TRPV1 expression profile in ccRCC has not been comprehensively elucidated. In this study, TRPV1 expression in ccRCC and other cancer types was analyzed based on data from the GEO and Oncomine databases. Immunohistochemical (IHC) staining was performed for further validation in human ccRCC tissue chips. Survival and correlation analyses of TRPV1 were conducted using Kaplan-Meier Plotter (KM-Plotter) and the Tumor IMmune Estimation Resource (TIMER) database. TRPV1 exhibited a low expression profile in 2 GEO datasets (GSE6344, GSE36895) and 4 Oncomine datasets (Gumz, Lenburg, Beroukhim 1 and Beroukhim 2), as also confirmed by IHC staining. Survival analysis indicated that high enrichment of TRPV1 significantly predicted a better overall survival (OS) and disease-free survival (DFS) of 1, 3, 5 and 10 years in ccRCC patients. TIMER analysis showed that TRPV1 copy number alterations (CNA) were closely related to immune cell infiltration. The detailed results indicated that TRPV1 expression was positively correlated with the infiltration level of CD4+ T cells, but negatively correlated with B cells, macrophages, and dendritic cells infiltration. In addition, TRPV1 might also be inversely related to abundance of the regulatory T cells (Treg) and the M2 subset of macrophages. Finally, we found that TRPV1 expression was tightly associated with several key molecules of the classical pathways in ccRCC, such as VHL, TP53, HIF1A, MTOR, MAPK1, MET, CTNNB1, etc. Our research work suggests that TRPV1 is a novel tumor suppressor and prognosis marker for ccRCC and is of great value for further exploration.

Hashimoto thyroiditis (HT) is one of the most common autoimmune diseases, and the incidence of HT continues to increase. Long-term, uncontrollable HT results in thyroid dysfunction and even increases carcinogenesis risks. Since the origin and development of HT involve many complex immune processes, there is no effective therapy for HT on a pathogenesis level. Although bioinformatics analysis has been utilized to seek key genes and pathways of thyroid cancer, few bioinformatics studies that focus on HT pathogenesis and mechanisms have been reported. In the present study, the Gene Expression Omnibus (GEO) dataset (GSE29315) containing 6 HT and 8 thyroid physiological hyperplasia (TPH) samples was downloaded, and differentially expressed gene (DEG) analysis, Gene Ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, protein-protein interaction (PPI) analysis, and gene set enrichment analysis (GSEA) were performed. In total, 85 DEGs, containing 76 upregulated and 9 downregulated DEGS, were identified. The DEGs were mainly enriched in immune and inflammatory response, and the signaling pathways were involved in cytokine interaction and cytotoxicity. Moreover, ten hub genes were identified, and IFN-γ, IFN-α, IL6/JAK/STAT3 and inflammatory pathways may promote the origin and progression of HT. The present studies indicated that exploring DEGs and pathways by bioinformatics analysis has important significance in understanding the molecular mechanisms of HT and providing potential targets for the prevention and treatment of HT.

Porous titanium scaffold with suitable porous architecture exhibits enormous potentials for bone defect repairs. However, insufficient osteointegration and osteoinduction still remain to open as one of the major problems to achieve satisfactory therapeutic effect. To solve this problem, many studies have been carried out to improve the bioactivity of porous titanium scaffold by surface modifications. In this study, porous Ti6Al4V scaffolds were fabricated using additive manufacturing technique. Porous architectures were built up based on a diamond pore structure unit. Alkali-acid-heat (AH) treatment was applied to create a TiO 2 layer on the porous Ti6Al4V scaffold (AH-porous Ti6Al4V). Subsequently, a hydrothermal treatment was employed to enable the formation of HA coating with nanopillar-like morphology on the alkali-acid-heat-treated surface (HT/AH-porous Ti6Al4V). The effects of surface modifications on apatite-forming ability, protein adsorption, cell attachment, cell proliferation and osteogenic gene expression were studied using apatite-forming ability test, protein adsorption assay and in vitro cell culture assay. It was found that the HT/AH-porous Ti6Al4V exhibited the highest apatite formation ability and best affinity to fibronectin and vitronectin. In vitro studies indicated that the mesenchymal stem cells (MSCs) cultured on the HT/AH-porous Ti6Al4V presented improved adhesion and differentiation compared with the porous Ti6Al4V and AH-porous Ti6Al4V.

Osteosarcoma is a type of highly aggressive bone tumor arising from primitive cells of mesenchymal origin in adults and is associated with a high rate of tumor relapse. However, there is an urgent need to clarify the molecular mechanisms underlying osteosarcoma development. The present study performed integrated bioinformatics analysis in a single-cell RNA sequencing dataset and explored the potential interactive signaling pathways associated with osteosarcoma development. Single-cell transcriptomic analysis of osteosarcoma tissues was performed by using the Seurat R package, the Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of differentially expressed genes was performed by using the clusterProfiler R package, and the cell–cell interaction analysis was performed by using the CellPhoneDB package. Our results showed that 11 clustered cell types were identified across 11 osteosarcoma tissues, with cell types including “osteoblastic”, “myeloid”, “osteoblastic_proli”, “osteoclast”, and “tumor-infiltrating lymphocytes (TILs)” as the main types. The DEGs between different cell types from primary, metastatic, and recurrent osteosarcomas were mainly enriched in the GO terms including “negative regulation of hydrolase activity”, “regulation of peptidase activity”, “regulation of binding”, “negative regulation of proteolysis”, and “negative regulation of peptidase activity” and in the KEGG pathways including “transcriptional misregulation in cancer”, “cellular senescence”, “apoptosis”, “FoxO signaling pathway”, “cell cycle”, “NF-kappa B signaling pathway”, “p53 signaling pathway”, “pentose phosphate pathway”, and “protein export”. For the cell–cell communication network analysis, the different interaction profiles between cell types were detected among primary, metastatic, and recurrent osteosarcomas. Further exploration of the KEGG pathway revealed that these ligand/receptor interactions may be associated with the NF-κB signaling pathway and its interacted mediators. In conclusion, the present study for the first time explored the scRNA-seq dataset in osteosarcoma, and our results revealed the 11 clustered cell types and demonstrated the novel cell–cell interactions among different cell types in primary, metastatic, and recurrent osteosarcomas. The NF-κB signaling pathway may play a key role in regulating the TME of osteosarcoma. The present study may provide new insights into understanding the molecular mechanisms of osteosarcoma pathophysiology.

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