Ensuring a sufficient oxygen supply is pivotal for the success of bioprinting applications since it fosters tissue integration and natural regeneration.Variation in oxygen concentration among diverse tissues necessita...Ensuring a sufficient oxygen supply is pivotal for the success of bioprinting applications since it fosters tissue integration and natural regeneration.Variation in oxygen concentration among diverse tissues necessitates the precise recreation of tissue-specific oxygen levels in imprinted constructs to support the survival of targeted cells.Although oxygen-releasing biomaterials,such as oxygen-generating microparticles(OMPs),have shown promise for enhancing the oxygen supply of microenvironments in injured tissues,whether this approach is scalable for large tissues and whether tissue-specific bioinks with varying OMP concentrations remain printable remain unknown.This study addresses this critical gap by introducing an innovative class of engineered oxygenated bioinks that combine colloidal-based microgels with OMPs.We report that incorporating nanosized calcium peroxide(nCaO_(2))and manganese oxide nanosheets(nMnO_(2))into hydrophobic polymeric microparticles enables precise modulation of oxygen release while controlling hydrogen peroxide release.Moreover,the fabrication of oxygenating and cytocompatible colloidal gels is achieved using an aqueous two-phase system.This study thoroughly evaluates the fundamental characteristics of the resulting bioink,including its rheological behaviors,printability,shape fidelity,mechanical properties,and oxygen release properties.Moreover,this study demonstrates the macroscopic scalability and cytocompatibility of printed constructs produced via cell-laden oxygenating colloidal bioinks.By showcasing the effectiveness of extrusion-based bioprinting,this study underscores how it can be used to fabricate biomimetic tissues,indicating its potential for new applications.The findings presented here advance the bioprinting field by achieving scalability with both high cell viability and the possibility of mimicking specifically oxygenated tissues.This work thereby offers a promising avenue for the development of functional tissues with enhanced physiological relevance.展开更多
Since the start of the Precision Medicine Initiative by the United States of America in 2015,interest in personalized medicine has grown extensively.In short,personalized medicine is a term that describes medical trea...Since the start of the Precision Medicine Initiative by the United States of America in 2015,interest in personalized medicine has grown extensively.In short,personalized medicine is a term that describes medical treatment that is tuned to the individual.One possible way to realize personalized medicine is 3D printing.When using materials that can be tuned upon stimulation,4D printing is established.In recent years,many studies have been exploring a new field that combines 3D and 4D printing with therapeutics.This has resulted in many concepts of pharmaceutical devices and formulations that can be printed and,possibly,tailored to an individual.Moreover,the first 3D printed drug,Spritam®,has already found its way to the clinic.This review gives an overview of various 3D and 4D printing techniques and their applications in the pharmaceutical field as drug delivery systems and personalized medicine.展开更多
During liver injury,intrahepatic macrophage compartment is augmented by circulating monocytes that infiltrate the liver driven by C-C motif chemokine ligand/C-C motif chemokine receptor(CCL/CCR)axis including CCL1‒CCR...During liver injury,intrahepatic macrophage compartment is augmented by circulating monocytes that infiltrate the liver driven by C-C motif chemokine ligand/C-C motif chemokine receptor(CCL/CCR)axis including CCL1‒CCR8 axis,thereby contributing to liver inflammation.Numerous small molecular receptor antagonists,including R243,have been developed for targeting CCR8;however,these agents face challenges in clinical translation,potentially attributed to their poor pharmacokinetic profiles,lack of target specificity,and potential adverse effects.In this study,we designed four CCR8 antagonizing peptides(AP8i-AP8iv)and performed molecular characterization in silico and therapeutic investigation in vitro and in vivo.Based on in silico docking,molecular dynamic simulation using homology build model and in-vitro(competitive)binding studies,AP8ii(YEWRFYHG)evidenced highly favorable and selective interactions at the CCR8-active site.AP8ii inhibited CCL1-driven chemotaxis and LPS/IFNg-induced pro-inflammatory activation of monocytes-macrophages in vitro.In a CCl4-induced acute liver injury mouse model,AP8ii treatment decreased intrahepatic infiltration of circulating monocytes.Moreover,AP8ii reduced liver inflammation,as indicated by decreased F4/80,IL6 and iNOS expression,diminished ALT levels,and attenuated fibrosis,as indicated by reduced collagen-I expression.In conclusion,we report a novel CCR8-antagonizing peptide that inhibited CCL1-driven intrahepatic monocytes infiltration and differentiation into pro-inflammatory phenotype,consequently ameliorating liver inflammation and fibrogenesis in an acute liver injury mouse model.展开更多
Living microtissues are used in a multitude of applications as they more closely resemble native tissue physiology,as compared to 2D cultures.Microtissues are typically composed of a combination of cells and materials...Living microtissues are used in a multitude of applications as they more closely resemble native tissue physiology,as compared to 2D cultures.Microtissues are typically composed of a combination of cells and materials in varying combinations,which are dictated by the applications’design requirements.Their applications range wide,from fundamental biological research such as differentiation studies to industrial applications such as cruelty-free meat production.However,their translation to industrial and clinical settings has been hindered due to the lack of scalability of microtissue production techniques.Continuous microfluidic processes provide an opportunity to overcome this limitation as they offer higher throughput production rates as compared to traditional batch techniques,while maintaining reproducible control over microtissue composition and size.In this review,we provide a comprehensive overview of the current approaches to engineer microtissues with a focus on the advantages of,and need for,the use of continuous processes to produce microtissues in large quantities.Finally,an outlook is provided that outlines the required developments to enable large-scale microtissue fabrication using continuous processes.展开更多
A combination of the viscoelastic properties of hyaluronic acid(HA)and the elastic properties of star shaped 8-arm poly(ethylene glycol)(8-arm PEG)was used to design in-situ forming hydrogels.Hydrogels were prepared b...A combination of the viscoelastic properties of hyaluronic acid(HA)and the elastic properties of star shaped 8-arm poly(ethylene glycol)(8-arm PEG)was used to design in-situ forming hydrogels.Hydrogels were prepared by the enzymatic crosslinking of a partially tyramine modified 8-arm PEG and a tyramine conjugated HA using horseradish peroxidase in the presence of hydrogen peroxide.Hydrogels of the homopolymer conjugates and mixtures thereof were rapidly formed within seconds under physiological conditions at low polymer and enzyme concentrations.Elastic hydrogels with high gel content(≥95%)and high storage moduli(up to 22.4 kPa)were obtained.An in vitro study in the presence of hyaluronidase(100 U/mL)revealed that with increasing PEG content the degradation time of the hybrid hydrogels increased up to several weeks,whereas hydrogels composed of only hyaluronic acid degraded within 2 weeks.Human mesenchymal stem cells(hMSCs)incorporated in the hybrid hydrogels remained viable as shown by a PrestoBlue and a live-dead assay,confirming the biocompatibility of the constructs.The production of an extracellular matrix by re-differentiation of encapsulated human chondrocytes was followed over a period of 28 days.Gene expression indicated that these highly elastic hydrogels induced an enhanced production of collagen type II.At low PEG-TA/HA-TA ratios a higher expression of SOX 9 and ACAN was observed.These results indicate that by modulating the ratio of PEG/HA,injectable hydrogels can be prepared applicable as scaffolds for tissue regeneration applications.展开更多
基金funded by the National Insti-tutes of Health(No.R01 AR074234)AHA collaborative award(No.944227)the Gillian Reny Stepping Strong Center for Trauma Inno-vation at Brigham and Women's Hospital.
文摘Ensuring a sufficient oxygen supply is pivotal for the success of bioprinting applications since it fosters tissue integration and natural regeneration.Variation in oxygen concentration among diverse tissues necessitates the precise recreation of tissue-specific oxygen levels in imprinted constructs to support the survival of targeted cells.Although oxygen-releasing biomaterials,such as oxygen-generating microparticles(OMPs),have shown promise for enhancing the oxygen supply of microenvironments in injured tissues,whether this approach is scalable for large tissues and whether tissue-specific bioinks with varying OMP concentrations remain printable remain unknown.This study addresses this critical gap by introducing an innovative class of engineered oxygenated bioinks that combine colloidal-based microgels with OMPs.We report that incorporating nanosized calcium peroxide(nCaO_(2))and manganese oxide nanosheets(nMnO_(2))into hydrophobic polymeric microparticles enables precise modulation of oxygen release while controlling hydrogen peroxide release.Moreover,the fabrication of oxygenating and cytocompatible colloidal gels is achieved using an aqueous two-phase system.This study thoroughly evaluates the fundamental characteristics of the resulting bioink,including its rheological behaviors,printability,shape fidelity,mechanical properties,and oxygen release properties.Moreover,this study demonstrates the macroscopic scalability and cytocompatibility of printed constructs produced via cell-laden oxygenating colloidal bioinks.By showcasing the effectiveness of extrusion-based bioprinting,this study underscores how it can be used to fabricate biomimetic tissues,indicating its potential for new applications.The findings presented here advance the bioprinting field by achieving scalability with both high cell viability and the possibility of mimicking specifically oxygenated tissues.This work thereby offers a promising avenue for the development of functional tissues with enhanced physiological relevance.
文摘Since the start of the Precision Medicine Initiative by the United States of America in 2015,interest in personalized medicine has grown extensively.In short,personalized medicine is a term that describes medical treatment that is tuned to the individual.One possible way to realize personalized medicine is 3D printing.When using materials that can be tuned upon stimulation,4D printing is established.In recent years,many studies have been exploring a new field that combines 3D and 4D printing with therapeutics.This has resulted in many concepts of pharmaceutical devices and formulations that can be printed and,possibly,tailored to an individual.Moreover,the first 3D printed drug,Spritam®,has already found its way to the clinic.This review gives an overview of various 3D and 4D printing techniques and their applications in the pharmaceutical field as drug delivery systems and personalized medicine.
文摘During liver injury,intrahepatic macrophage compartment is augmented by circulating monocytes that infiltrate the liver driven by C-C motif chemokine ligand/C-C motif chemokine receptor(CCL/CCR)axis including CCL1‒CCR8 axis,thereby contributing to liver inflammation.Numerous small molecular receptor antagonists,including R243,have been developed for targeting CCR8;however,these agents face challenges in clinical translation,potentially attributed to their poor pharmacokinetic profiles,lack of target specificity,and potential adverse effects.In this study,we designed four CCR8 antagonizing peptides(AP8i-AP8iv)and performed molecular characterization in silico and therapeutic investigation in vitro and in vivo.Based on in silico docking,molecular dynamic simulation using homology build model and in-vitro(competitive)binding studies,AP8ii(YEWRFYHG)evidenced highly favorable and selective interactions at the CCR8-active site.AP8ii inhibited CCL1-driven chemotaxis and LPS/IFNg-induced pro-inflammatory activation of monocytes-macrophages in vitro.In a CCl4-induced acute liver injury mouse model,AP8ii treatment decreased intrahepatic infiltration of circulating monocytes.Moreover,AP8ii reduced liver inflammation,as indicated by decreased F4/80,IL6 and iNOS expression,diminished ALT levels,and attenuated fibrosis,as indicated by reduced collagen-I expression.In conclusion,we report a novel CCR8-antagonizing peptide that inhibited CCL1-driven intrahepatic monocytes infiltration and differentiation into pro-inflammatory phenotype,consequently ameliorating liver inflammation and fibrogenesis in an acute liver injury mouse model.
基金financial support from Dutch Research Council(Vidi,17522)European Research Council(Starting Grant,759425),European Fund for Regional Development(EFRO-00963)Dutch Arthritis Foundation(17-1-405).
文摘Living microtissues are used in a multitude of applications as they more closely resemble native tissue physiology,as compared to 2D cultures.Microtissues are typically composed of a combination of cells and materials in varying combinations,which are dictated by the applications’design requirements.Their applications range wide,from fundamental biological research such as differentiation studies to industrial applications such as cruelty-free meat production.However,their translation to industrial and clinical settings has been hindered due to the lack of scalability of microtissue production techniques.Continuous microfluidic processes provide an opportunity to overcome this limitation as they offer higher throughput production rates as compared to traditional batch techniques,while maintaining reproducible control over microtissue composition and size.In this review,we provide a comprehensive overview of the current approaches to engineer microtissues with a focus on the advantages of,and need for,the use of continuous processes to produce microtissues in large quantities.Finally,an outlook is provided that outlines the required developments to enable large-scale microtissue fabrication using continuous processes.
基金support by a grant from the Dutch government to the Netherlands Institute for Regenerative Medicine(NIRM,Grant No.FES0908)support of the Netherlands Organisation for Scientific Research(NWO)P15-23(Project 1)“Activating resident stem cells”and EFRO OPOost 00867 Orthros TR.
文摘A combination of the viscoelastic properties of hyaluronic acid(HA)and the elastic properties of star shaped 8-arm poly(ethylene glycol)(8-arm PEG)was used to design in-situ forming hydrogels.Hydrogels were prepared by the enzymatic crosslinking of a partially tyramine modified 8-arm PEG and a tyramine conjugated HA using horseradish peroxidase in the presence of hydrogen peroxide.Hydrogels of the homopolymer conjugates and mixtures thereof were rapidly formed within seconds under physiological conditions at low polymer and enzyme concentrations.Elastic hydrogels with high gel content(≥95%)and high storage moduli(up to 22.4 kPa)were obtained.An in vitro study in the presence of hyaluronidase(100 U/mL)revealed that with increasing PEG content the degradation time of the hybrid hydrogels increased up to several weeks,whereas hydrogels composed of only hyaluronic acid degraded within 2 weeks.Human mesenchymal stem cells(hMSCs)incorporated in the hybrid hydrogels remained viable as shown by a PrestoBlue and a live-dead assay,confirming the biocompatibility of the constructs.The production of an extracellular matrix by re-differentiation of encapsulated human chondrocytes was followed over a period of 28 days.Gene expression indicated that these highly elastic hydrogels induced an enhanced production of collagen type II.At low PEG-TA/HA-TA ratios a higher expression of SOX 9 and ACAN was observed.These results indicate that by modulating the ratio of PEG/HA,injectable hydrogels can be prepared applicable as scaffolds for tissue regeneration applications.
