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1.
Biomed Mater Eng ; 28(s1): S75-S79, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28372280

RESUMO

Mesenchymal stem cells (MSCs) are a common tool in regenerative medicine. The nanoscale extracellular vesicles (nEVs) secreted by these cells were recently brought up to light thanks to their therapeutic potential. In this study, we assessed the in vitro behaviour of human umbilical vein endothelial cells (HUVECs) exposed to nEVs derived from human umbilical cord mesenchymal stem cells (hUC-MSCs). Nanoscale extracellular vesicles were isolated and characterized by NanoSight® and flow cytometry. HUVECs were stimulated with various concentrations of nEVs. To assess nEV interactions with HUVECs, confocal microscopy and angiogenesis assay were performed. The use of nEVs derived from hUC-MSCs was able to produce positive outcomes on HUVECs by acting on their angiogenic potential.


Assuntos
Indutores da Angiogênese/metabolismo , Vesículas Extracelulares/metabolismo , Células-Tronco Mesenquimais/citologia , Neovascularização Fisiológica , Cordão Umbilical/citologia , Técnicas de Cultura de Células , Células Cultivadas , Vesículas Extracelulares/ultraestrutura , Células Endoteliais da Veia Umbilical Humana , Humanos , Células-Tronco Mesenquimais/metabolismo , Tamanho da Partícula
2.
J Struct Biol ; 196(2): 127-137, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-26994554

RESUMO

Nacre is able to induce bone-forming cells mineralization, and gains widely interest in bone regeneration. While, the osteoinductive compounds are not yet identified. ESM (Ethanol Soluble Matrix), a nacre extract from powder of Pinctada margaritifera pearl oyster shell, has been firstly proven having the capacity to induce mineralization and to restore mineralization defect in vitro. It is suitable to treat ESM as a source of osteoinductive compounds. Herein, we develop a new method for separating and purifying nacre extracts by an ionic approach. At first, cationic ESM (ESMc) and anionic ESM (ESMa) were achieved with ion-exchange resin. Then, ESM was separated and collected on cation exchange HPLC. Scanning Electron Microscopy coupled with Energy Dispersive X-ray Spectrometry (EDS) was used to reveal the concentrated elements in ESM fractions. A coupled cell models were used to test the ESM fractions. Alizarin Red staining was performed and quantified to evaluate the mineralization level. ESMc and 2 HPLC fractions stimulated the mineralization in both cells. EDS demonstrated the abundant presence of calcium and chloride in the osteogenic fractions. To validate, pure CaCl2 was tested and proven having an osteogenic effect in both cells, but less stable than ESM. The mineralization nodules induced by ESM fractions and CaCl2 differed in both cells. In conclusion, a new method was developed for separating and purifying nacre extracts by an ionic approach. By which, the osteoinductive compounds in ESM were proven cationic, and calcium in ESM was demonstrated to play a role in inducing the cell mineralization.


Assuntos
Calcificação Fisiológica/efeitos dos fármacos , Nácar/química , Nácar/farmacologia , Osteogênese/efeitos dos fármacos , Células 3T3/efeitos dos fármacos , Animais , Cátions , Etanol , Humanos , Camundongos , Nácar/isolamento & purificação , Osteoblastos/citologia , Osteoblastos/efeitos dos fármacos , Pinctada/química
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