The highly organized extracellular matrix(ECM)of musculoskeletal tissues,encompassing tendons,ligaments and muscles,is structurally anisotropic,hierarchical and multi-compartmental.These features collectively contribu...The highly organized extracellular matrix(ECM)of musculoskeletal tissues,encompassing tendons,ligaments and muscles,is structurally anisotropic,hierarchical and multi-compartmental.These features collectively contribute to their unique function.Previous studies have investigated the effect of tissue-engineered scaffold anisotropy on cell morphology and organization for musculoskeletal tissue repair and regeneration,but the hierarchical arrangement of ECM and compartmentalization are not typically replicated.Here,we present a method for multi-compartmental scaffold design that allows for physical mimicry of the spatial architecture of musculoskeletal tissue in regenerative medicine.This design is based on an ECM-inspired macromolecule scaffold.Polycaprolactone(PCL)scaffolds were fabricated with aligned fibers by electrospinning and mechanical stretching,and then surface-functionalized with the cell-supporting ECM protein molecule,tropoelastin(TE).TE was attached using two alternative methods that allowed for either physisorption or covalent attachment,where the latter was achieved by plasma ion immersion implantation(PIII).Aligned fibers stimulated cell elongation and improved cell alignment,in contrast to randomly oriented fibers.TE coatings bound by physisorption or covalently following 200 s PIII treatment promoted fibroblast proliferation.This represents the first cytocompatibility assessment of novel PIII-treated TE-coated PCL scaffolds.To demonstrate their versatility,these 2D anisotropic PCL scaffolds were assembled into 3D hierarchical constructs with an internally compartmentalized structure to mimic the structure of musculoskeletal tissue.展开更多
Many biological structures such as nerves,blood and lymphatic vessels,and muscle fibres exhibit longitudinal ge-ometries with distinct cell types extending along both the length and width of internal linear axes.Model...Many biological structures such as nerves,blood and lymphatic vessels,and muscle fibres exhibit longitudinal ge-ometries with distinct cell types extending along both the length and width of internal linear axes.Modelling these three-dimensional structures in vitro is challenging:the best-defined stem-cell differentiation systems are mono-layer cultures or organoids using pluripotent stem cells.Pluripotent stem cells can differentiate into functionally mature cells depending on the signals received,holding great promise for regenerative medicine.However,the integration of in vitro differentiated cell types into diseased tissue remains a challenge.Engineered scaffolds can bridge this gap if the appropriate signalling systems are incorporated into the scaffold.Here,we have taken a biomimicry approach to generate longitudinal structures in vitro.In this approach,mouse embryonic stem cells are directed to differentiate to specific cell types on the surface of polycaprolactone(PCL)fibres treated by plasma-immersion ion implantation and to which with lineage-specifying molecules have been covalently im-mobilised.We demonstrate the simplicity and utility of our method for efficiently generating high yields of the following cell types from these pluripotent stem cells:neurons,vascular endothelial cells,osteoclasts,adipocytes,and cells of the erythroid,myeloid,and lymphoid lineages.Strategically arranged plasma-treated scaffolds with differentiated cell types could ultimately serve as a means for the repair or treatment of diseased or damaged tissue.展开更多
基金supported by an Australian Commonwealth Government Research Training Program Tuition Fee Offset and Stipend Scholarship.A.S.W.acknowledges funding from the National Health and Medical Research Council(APP1195827)M.M.M.B.acknowledges funding from the Australian Research Council(FL190100216).
文摘The highly organized extracellular matrix(ECM)of musculoskeletal tissues,encompassing tendons,ligaments and muscles,is structurally anisotropic,hierarchical and multi-compartmental.These features collectively contribute to their unique function.Previous studies have investigated the effect of tissue-engineered scaffold anisotropy on cell morphology and organization for musculoskeletal tissue repair and regeneration,but the hierarchical arrangement of ECM and compartmentalization are not typically replicated.Here,we present a method for multi-compartmental scaffold design that allows for physical mimicry of the spatial architecture of musculoskeletal tissue in regenerative medicine.This design is based on an ECM-inspired macromolecule scaffold.Polycaprolactone(PCL)scaffolds were fabricated with aligned fibers by electrospinning and mechanical stretching,and then surface-functionalized with the cell-supporting ECM protein molecule,tropoelastin(TE).TE was attached using two alternative methods that allowed for either physisorption or covalent attachment,where the latter was achieved by plasma ion immersion implantation(PIII).Aligned fibers stimulated cell elongation and improved cell alignment,in contrast to randomly oriented fibers.TE coatings bound by physisorption or covalently following 200 s PIII treatment promoted fibroblast proliferation.This represents the first cytocompatibility assessment of novel PIII-treated TE-coated PCL scaffolds.To demonstrate their versatility,these 2D anisotropic PCL scaffolds were assembled into 3D hierarchical constructs with an internally compartmentalized structure to mimic the structure of musculoskeletal tissue.
基金supported by the Australian Research Council Laureate and Discovery fundings[FL190100216,DP190103507 and DE210100662]the University of Sydney School of Physics“Grand Challenge”program.
文摘Many biological structures such as nerves,blood and lymphatic vessels,and muscle fibres exhibit longitudinal ge-ometries with distinct cell types extending along both the length and width of internal linear axes.Modelling these three-dimensional structures in vitro is challenging:the best-defined stem-cell differentiation systems are mono-layer cultures or organoids using pluripotent stem cells.Pluripotent stem cells can differentiate into functionally mature cells depending on the signals received,holding great promise for regenerative medicine.However,the integration of in vitro differentiated cell types into diseased tissue remains a challenge.Engineered scaffolds can bridge this gap if the appropriate signalling systems are incorporated into the scaffold.Here,we have taken a biomimicry approach to generate longitudinal structures in vitro.In this approach,mouse embryonic stem cells are directed to differentiate to specific cell types on the surface of polycaprolactone(PCL)fibres treated by plasma-immersion ion implantation and to which with lineage-specifying molecules have been covalently im-mobilised.We demonstrate the simplicity and utility of our method for efficiently generating high yields of the following cell types from these pluripotent stem cells:neurons,vascular endothelial cells,osteoclasts,adipocytes,and cells of the erythroid,myeloid,and lymphoid lineages.Strategically arranged plasma-treated scaffolds with differentiated cell types could ultimately serve as a means for the repair or treatment of diseased or damaged tissue.
