RESUMO
BACKGROUND: Alcohol abuse is known to be a leading risk factor for atraumatic osteonecrosis of the femoral head (ONFH), in which the suppression of osteogenesis plays a critical role. Cordycepin benefits bone metabolism; however, there has been no study to determine its effect on osteonecrosis. METHODS: Human bone mesenchymal stem cells (hBMSCs) were identified by multi-lineage differentiation. Alkaline phosphatase (ALP) activity, RT-PCR, western blots, immunofluorescent assay and Alizarin red staining of BMSCs were evaluated. A rat model of alcohol-induced ONFH was established to investigate the protective role of cordycepin against ethanol. Hematoxylin & eosin (H&E) staining and micro-computerized tomography (micro-CT) were performed to observe ONFH. Apoptosis was assessed by TdT-mediated dUTP nick end labeling (TUNEL). Immunohistochemical staining was carried out to detect OCN and COL1. RESULTS: Ethanol significantly suppressed ALP activity, decreased gene expression of OCN and BMP2, lowered levels of RUNX2 protein, and reduced immunofluorescence staining of OCN and COL1 and calcium formation of hBMSCs. However, these inhibitory effects were attenuated by cordycepin co-treatment at concentrations of 1 and 10 µg/mL Moreover, it was revealed that the osteo-protective effect of cordycepin was associated with modulation of the Wnt/ß-catenin pathway. In vivo, by micro-CT, TUNEL and immunohistochemical staining of OCN and COL1, we found that cordycepin administration prevented alcohol-induced ONFH. CONCLUSION: Cordycepin treatment to enhance osteogenesis may be considered a potential therapeutic approach to prevent the development of alcohol-induced ONFH.
Assuntos
Diferenciação Celular/efeitos dos fármacos , Desoxiadenosinas/farmacologia , Etanol/toxicidade , Osteogênese/efeitos dos fármacos , Substâncias Protetoras/farmacologia , Animais , Proteína Morfogenética Óssea 2/genética , Proteína Morfogenética Óssea 2/metabolismo , Células Cultivadas , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Subunidade alfa 1 de Fator de Ligação ao Core/genética , Subunidade alfa 1 de Fator de Ligação ao Core/metabolismo , Cabeça do Fêmur/diagnóstico por imagem , Cabeça do Fêmur/patologia , Colo do Fêmur/citologia , Humanos , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Modelos Animais , Osteocalcina/genética , Osteocalcina/metabolismo , Ratos , Ratos Sprague-Dawley , Via de Sinalização Wnt/efeitos dos fármacos , beta Catenina/metabolismoRESUMO
An Au(i)-catalyzed cyclization/1,2-rearrangement/aromatization cascade of 3-yne-1,2-diols has been successfully realized. This reaction not only provides a new and efficient strategy for the synthesis of substituted cycloalka[b]furan compounds as well as their derivatives, but might also facilitate related biological studies.
RESUMO
Alcohol is considered a leading risk factor for osteopenia. Our previous research indicated that the Akt/GSK-3ß/ß-catenin pathway plays a critical role in the ethanol-induced antiosteogenic effect in bone mesenchymal stem cells (BMSCs). PI3K/Akt is negatively regulated by the phosphatase and tensin homolog (PTEN) phosphatase. In this study, we found that ethanol increased PTEN expression in the BMSCs and bone tissue of ethanol-treated Sprague-Dawley rats. PTEN upregulation impaired Akt recruitment to the plasma membrane and suppressed Akt phosphorylation at Ser473, thereby inhibiting Akt/GSK3ß/ß-catenin signaling and the expression of COL1 and OCN in BMSCs in vitro and in vivo. The results of in vivo assays indicated that PTEN inhibition protected bone tissue against ethanol. Interestingly, our data revealed that following ethanol stimulation, PTEN and PTEN pseudogene 1 (PTENP1) mRNA expression was increased in a time-dependent manner, resulting in an increased PTEN protein level. In addition, ethanol upregulated PTEN expression and decreased PTEN phosphorylation (p-PTEN), indicating an increase in functional PTEN levels. In summary, the ethanol-mediated transcriptional and post-transcriptional regulation of PTEN impaired downstream Akt/GSK3ß/ß-catenin signaling and BMSC osteogenic differentiation. Therefore, we propose that Akt/GSK3ß/ß-catenin activation via PTEN inhibition may be a potential therapeutic approach for preventing the development of alcohol-induced osteopenia.
Assuntos
Doenças Ósseas Metabólicas/patologia , Membrana Celular/metabolismo , Etanol/toxicidade , Glicogênio Sintase Quinase 3 beta/metabolismo , PTEN Fosfo-Hidrolase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , beta Catenina/metabolismo , Animais , Doenças Ósseas Metabólicas/induzido quimicamente , Doenças Ósseas Metabólicas/metabolismo , Membrana Celular/efeitos dos fármacos , Células Cultivadas , Depressores do Sistema Nervoso Central/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Glicogênio Sintase Quinase 3 beta/genética , Masculino , Células-Tronco Mesenquimais/citologia , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , PTEN Fosfo-Hidrolase/genética , Proteínas Proto-Oncogênicas c-akt/genética , Ratos , Ratos Sprague-Dawley , beta Catenina/genéticaRESUMO
Bone repair materials for the effective treatment of bone defects should simultaneously possess excellent biocompatibility and promote osteogenic differentiation. Herein, we prepared pifithrin-α-loaded layered double hydroxide/chitosan (PFTα-LDH-CS) nanohybrid composites for the first time according to the following steps: (i) the immersion of LDH nanoplates and PFTα in a CS solution; and (ii) the self-assembly synthesis of PFTα-LDH-CS nanohybrid composites after the pH value of the mixed solution was adjusted to 7.4. Interestingly, the LDH nanoplates with a thickness of â¼20 nm and width of â¼300 nm agglomerated together into flower-like shapes by self-assembly, and the CS was dispersed around the LDH nanoplates. The mesopores with the pore size of 3.95 nm among the LDH nanoplates served as channels for loading PFTα. Moreover, the CS around the LDH nanoplates increased the drug loading efficiency and drug sustained release property compared with the pure LDH nanoplates. The in vitro tests demonstrated that the human bone marrow-derived mesenchymal stem cells (hBMSCs) had good adhesion, spreading and proliferating on the LDH-CS and PFTα-LDH-CS, suggesting that both samples had the desired cytocompatibility. Note that the PFTα released from the PFTα-LDH-CS rapidly improved the cell proliferation, ALP activity, ECM mineralization and protein level of the Runt-related transcription factor 2 (RUNX2) and ß-catenin. The enhanced osteogenic differentiation of hBMSCs on the PFTα-LDH-CS may be attributed to the PFTα released from the abovementioned nanohybrid composites, which resulted in the accumulation of ß-catenin and activation of the ß-catenin-mediated transcription activity in the cell nucleus. Therefore, the PFTα-LDH-CS nanohybrid composites with excellent cytocompatibility and enhanced osteoinductivity have great applications for novel bone repair materials.
RESUMO
Alcohol is a leading risk factor for osteonecrosis of the femoral head (ONFH). We explored the molecular mechanisms underlying alcohol-induced ONFH and investigated the protective effect of the novel Akt activator SC-79 against this disease. We found that ethanol inhibited expression of the osteogenic genes RUNX2 and OCN, downregulated osteogenic differentiation, impaired the recruitment of Akt to the plasma membrane, and suppressed Akt phosphorylation at Ser473, thereby inhibiting the Akt/GSK3ß/ß-catenin signaling pathway in bone mesenchymal stem cells. To assess SC-79's ability to counteract the inhibitory effect of ethanol on Akt-Ser73 phosphorylation, we performed micro-computerized tomography and immunofluorescent staining of osteopontin, osteocalcin and collagen type 1 in a rat model of alcohol-induced ONFH. We found that SC-79 injections inhibited alcohol-induced osteonecrosis. These results show that alcohol-induced ONFH is associated with suppression of p-Akt-Ser473 in the Akt/GSK3ß/ß-catenin signaling pathway in bone mesenchymal stem cells. We propose that SC-79 treatment to rescue Akt activation could be tested in the clinic as a potential therapeutic approach to preventing the development of alcohol-induced ONFH.
Assuntos
Acetatos/farmacologia , Consumo de Bebidas Alcoólicas/efeitos adversos , Benzopiranos/farmacologia , Necrose da Cabeça do Fêmur/etiologia , Necrose da Cabeça do Fêmur/metabolismo , Proteínas Proto-Oncogênicas c-akt/agonistas , Proteínas Proto-Oncogênicas c-akt/metabolismo , Animais , Biópsia , Modelos Animais de Doenças , Necrose da Cabeça do Fêmur/diagnóstico , Necrose da Cabeça do Fêmur/tratamento farmacológico , Glicogênio Sintase Quinase 3 beta/antagonistas & inibidores , Glicogênio Sintase Quinase 3 beta/metabolismo , Humanos , Masculino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Osteogênese/efeitos dos fármacos , Ratos , Transdução de Sinais/efeitos dos fármacos , Microtomografia por Raio-X , beta Catenina/metabolismoRESUMO
An unprecedented nucleophilic addition/cyclization/aromatization cascade of basic chemicals, i.e., aromatic alkenes/alkynes, trimethylsilyl cyanide and N,N-disubstituted formamide, has been developed to give a series of multisubstituted pyrroles in moderate to good yields with high regioselectivities. This reaction not only reveals a new reaction mode for α-aminonitriles, but also provides a new and efficient cyclization pattern for the synthesis of multisubstituted pyrroles as well as their derivatives, which might facilitate related biological studies.
RESUMO
The properties of bone scaffolds, including biocompatibility, osteoinductivity and antibacterial activity, are of great importance for reconstruction of large bone defects and prevention of implant-associated infections. Herein, we develop an Ag-loaded strontium hydroxyapatite (SrHAP)/chitosan (CS) porous scaffold (Ag-SrHAP/CS) according to the following steps: (i) freeze-drying fabrication of a SrHAP/CS porous scaffold; and (ii) deposition of Ag nanoparticles on the above scaffold. In addition, HAP/CS and Ag-HAP/CS porous scaffolds are prepared under the same conditions without doping Sr element. All the HAP/CS, Ag-HAP/CS, SrHAP/CS and Ag-SrHAP/CS porous scaffolds provide a friendly environment for the adhesion, spreading and proliferation of human bone marrow mesenchymal stem cells (hBMSCs). The three-dimensional (3D) interconnected macropores with a pore size of 100-400 µm allow the spreading of hBMSCs throughout the whole scaffolds. Interestingly, the Sr ions and Ag ions released from the Ag-SrHAP/CS porous scaffolds significantly enhance their osteoinductivity and antibacterial activity, respectively. The Sr element in the SrHAP/CS and Ag-SrHAP/CS porous scaffolds increase the alkaline phosphatase (ALP) activity of hBMSCs, extracellular matrix (ECM) mineralization, and the expression levels of osteogenic-related genes BMP-2 and COL-I. Moreover, the Ag ions released from the Ag-HAP/CS and Ag-SrHAP/CS scaffolds can effectively inhibit the growth and attachment of Staphylococcus aureus (S. aureus, ATCC 25923). In conclusion, the Ag-SrHAP/CS porous scaffold possesses excellent biocompatibility, osteoinductivity and antibacterial activity, so it has great potential for application in bone tissue engineering to repair bone defects and avoid infections.
RESUMO
A novel SnCl4-promoted tandem reaction toward multisubstituted indenes via a sequential C-O bond cleavage/Nazarov cyclization/nucleophilic addition reaction has been developed to afford a series of multisubstituted indenes with an all-carbon quaternary center in moderate to good yields.
RESUMO
An Au(I)-catalyzed tandem reaction for the synthesis of 2-phenyl-1,4,5,6-tetrahydrocyclopenta[b]pyrrole derivatives from 1-(1-hydroxy-3-phenylprop-2-yn-1-yl)cyclobutanol and primary amines or NH4OAc has been developed to afford a series of polysubstituted pyrroles in moderate to good yields.