Amelogenin isoforms constitute the predominant component in the enamel matrix and each amelogenin isoform executes unique role in the enamel biomineralization process. Enamel matrix derivative enriching amelogenin iso...Amelogenin isoforms constitute the predominant component in the enamel matrix and each amelogenin isoform executes unique role in the enamel biomineralization process. Enamel matrix derivative enriching amelogenin isoforms have also hioactive property for tissue regeneration. Despite the development of recombinant protein technology that has greatly forwarded the understanding ofamelogenin properties, substantial evidences have revealed biochemical and functional difference between natural amelogenins and their recombinant form. To facilitate the study of enamel formation mechanism, more facile methodology to purify multiple natural amelogenin isoforms is pursued. Here we developed an effective one-shot method via reverse phase high-performance liquid chromatography （RP-HPLC） to purify various amelogenin isoforms from pig-derived amelogenin complex. A thorough process of chromatographic condition establishment including sample analysis on analytical scale and chromatographic condition design on preparative scale was described. Three representative amelogenin isoforms （TRAP, P148, P173） were isolated in one step and their purity was confirmed by matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and high resolution mass spectrometry.展开更多
Yeast cells have controllable biosorption on metallic ions during metabolism.However,few studies were dedicated to using yeast-regulated biomimetic mineralization process to control the strontium-doped positions in ca...Yeast cells have controllable biosorption on metallic ions during metabolism.However,few studies were dedicated to using yeast-regulated biomimetic mineralization process to control the strontium-doped positions in calcium phosphate microcapsules.In this study,the yeast cells were allowed to pre-adsorb strontium ions metabolically and then served as sacrificing template for the precipitation and calcination of mineral shell.The pre-adsorption enabled the microorganism to enrich of strontium ions into the inner part of the microcapsules,which ensured a slow-release profile of the trace element from the microcapsule.The co-culture with human marrow stromal cells showed that gene expressions of alkaline phosphatase and Collagen-I were promoted.The promotion of osteogenic differentiation was further confirmed in the 3D culture of cell-material complexes.The strategy using living microorganism as‘smart doping apparatus’to control incorporation of trace element into calcium phosphate paved a pathway to new functional materials for hard tissue regeneration.展开更多
基金financially supported by the National Natural Science Foundation of China (No. 51572087)the 111 Project (No. B13039)+1 种基金the Science and Technology Program of Guangdong Province (No. 2013B010403007)the Program for Changjiang Scholars and Innovative Research Team in University (IRT 0919)
文摘Amelogenin isoforms constitute the predominant component in the enamel matrix and each amelogenin isoform executes unique role in the enamel biomineralization process. Enamel matrix derivative enriching amelogenin isoforms have also hioactive property for tissue regeneration. Despite the development of recombinant protein technology that has greatly forwarded the understanding ofamelogenin properties, substantial evidences have revealed biochemical and functional difference between natural amelogenins and their recombinant form. To facilitate the study of enamel formation mechanism, more facile methodology to purify multiple natural amelogenin isoforms is pursued. Here we developed an effective one-shot method via reverse phase high-performance liquid chromatography （RP-HPLC） to purify various amelogenin isoforms from pig-derived amelogenin complex. A thorough process of chromatographic condition establishment including sample analysis on analytical scale and chromatographic condition design on preparative scale was described. Three representative amelogenin isoforms （TRAP, P148, P173） were isolated in one step and their purity was confirmed by matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and high resolution mass spectrometry.
基金This work was supported by National Basic Research Program of China(2012CB619100)National Natural Science Foundation of China(51072056,51572087)+2 种基金the 111 Project(B13039)Key grant of Chinese Ministry of Education(313022)Program for Changjiang Scholars and Innovative Research Team in University(IRT 0919).
文摘Yeast cells have controllable biosorption on metallic ions during metabolism.However,few studies were dedicated to using yeast-regulated biomimetic mineralization process to control the strontium-doped positions in calcium phosphate microcapsules.In this study,the yeast cells were allowed to pre-adsorb strontium ions metabolically and then served as sacrificing template for the precipitation and calcination of mineral shell.The pre-adsorption enabled the microorganism to enrich of strontium ions into the inner part of the microcapsules,which ensured a slow-release profile of the trace element from the microcapsule.The co-culture with human marrow stromal cells showed that gene expressions of alkaline phosphatase and Collagen-I were promoted.The promotion of osteogenic differentiation was further confirmed in the 3D culture of cell-material complexes.The strategy using living microorganism as‘smart doping apparatus’to control incorporation of trace element into calcium phosphate paved a pathway to new functional materials for hard tissue regeneration.
