A variety of engineered nanoparticles,including lipid nanoparticles,polymer nanoparticles,gold nanoparticles,and biomimetic nanoparticles,have been studied as delivery vehicles for biomedical applications.When assessi...A variety of engineered nanoparticles,including lipid nanoparticles,polymer nanoparticles,gold nanoparticles,and biomimetic nanoparticles,have been studied as delivery vehicles for biomedical applications.When assessing the efficacy of a nanoparticle-based delivery system,in vitro testing with a model delivery system is crucial because it allows for real-time,in situ quantitative transport analysis,which is often difficult with in vivo animal models.The advent of tissue engineering has offered methods to create experimental models that can closely mimic the 3D microenvironment in the human body.This review paper overviews the types of nanoparticle vehicles,their application areas,and the design strategies to improve delivery efficiency,followed by the uses of engineered microtissues and methods of analysis.In particular,this review highlights studies on multicellular spheroids and other 3D tissue engineering approaches for cancer drug development.The use of bio-engineered tissues can potentially provide low-cost,high-throughput,and quantitative experimental platforms for the development of nanoparticle-based delivery systems.展开更多
3D （Three-dimensional） Caco-2 spheroids closely recapitulating in vivo physiological organization of intestinal epithelial cells, provide an excellent in vitro model system to study their pathophysiology and their r...3D （Three-dimensional） Caco-2 spheroids closely recapitulating in vivo physiological organization of intestinal epithelial cells, provide an excellent in vitro model system to study their pathophysiology and their response to stressful stimuli. The objective of this technical note is to provide optimized in vitro experimental protocols for culturing 3D Caco-2 spheroids and for analyzing their cell growth features. An optimized 3D Caco-2 spheroid culturing technique based on a new configuration of the culture medium is provided A methodological approach to determine the distribution of the cell cycle phases in disaggregated Caco-2 spheroids by using cytofluorimetric analysis is also described. The optimized culturing protocol favors 3D Caco-2 spheroid differentiation process, as evaluated by the number of well-differentiated spheroids with a single hollow lumen. The cytofluorimetric analysis allows rapid collection of cell cycle phase data from high numbers of spheroid samples, thus, permitting to estimate their growth dynamics in a relatively short time. The optimized technical approaches described here can be applied in systematic manner to a variety of research activities utilizing 3D Caco-2 spheroids. Ease of use, time and economic saving advantages deriving from these protocols further highlight their potential.展开更多
基金NSF(CCSS-1809047,CAREER-1653702)NIH(1R01AR072027-01,1R03AR069383-01)the office of undergraduate research(OUR)at the University of Connecticut for providing funds to support this study.
文摘A variety of engineered nanoparticles,including lipid nanoparticles,polymer nanoparticles,gold nanoparticles,and biomimetic nanoparticles,have been studied as delivery vehicles for biomedical applications.When assessing the efficacy of a nanoparticle-based delivery system,in vitro testing with a model delivery system is crucial because it allows for real-time,in situ quantitative transport analysis,which is often difficult with in vivo animal models.The advent of tissue engineering has offered methods to create experimental models that can closely mimic the 3D microenvironment in the human body.This review paper overviews the types of nanoparticle vehicles,their application areas,and the design strategies to improve delivery efficiency,followed by the uses of engineered microtissues and methods of analysis.In particular,this review highlights studies on multicellular spheroids and other 3D tissue engineering approaches for cancer drug development.The use of bio-engineered tissues can potentially provide low-cost,high-throughput,and quantitative experimental platforms for the development of nanoparticle-based delivery systems.
文摘3D （Three-dimensional） Caco-2 spheroids closely recapitulating in vivo physiological organization of intestinal epithelial cells, provide an excellent in vitro model system to study their pathophysiology and their response to stressful stimuli. The objective of this technical note is to provide optimized in vitro experimental protocols for culturing 3D Caco-2 spheroids and for analyzing their cell growth features. An optimized 3D Caco-2 spheroid culturing technique based on a new configuration of the culture medium is provided A methodological approach to determine the distribution of the cell cycle phases in disaggregated Caco-2 spheroids by using cytofluorimetric analysis is also described. The optimized culturing protocol favors 3D Caco-2 spheroid differentiation process, as evaluated by the number of well-differentiated spheroids with a single hollow lumen. The cytofluorimetric analysis allows rapid collection of cell cycle phase data from high numbers of spheroid samples, thus, permitting to estimate their growth dynamics in a relatively short time. The optimized technical approaches described here can be applied in systematic manner to a variety of research activities utilizing 3D Caco-2 spheroids. Ease of use, time and economic saving advantages deriving from these protocols further highlight their potential.
