Dataset Information

Michael-Type Addition of Gelatin on Electrospun Nanofibrils for Self-Assembly of Cell Sheets Composed of Human Dermal Fibroblasts.

ABSTRACT: To facilitate cell sheet formation of human dermal fibroblasts, gelatin moieties were chemically decorated onto the surface of electrospun nanofibrils (NFs). Poly(caprolactone) [PCL] was electrospun onto fibrous meshes and then fragmented into nanofibrils by optimized milling and hydrolysis. After aminolysis of the NFs, methacrylated gelatin (GelMA) was reacted via Michael-type addition with the surface-exposed amines of the aminolyzed NFs (ahPCL NFs). GelMA was immobilized on the ahPCL NFs. Analysis of ahPCL NFs and native NFs conducted using X-ray photoelectron spectroscopy confirmed that gelatin was chemically conjugated onto the NFs. Human dermal fibroblasts (HDF) and the decorated NFs were self-assembled into cell sheets, and cells in the matrix showed highly spreading morphology by confocal microscopy. Our results indicate that the degree of cell spreading and cellular viability was much higher in the presence of GelMA immobilized in ahPCL NFs.

SUBMITTER: Lee JW

PROVIDER: S-EPMC6854572 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature

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Michael-Type Addition of Gelatin on Electrospun Nanofibrils for Self-Assembly of Cell Sheets Composed of Human Dermal Fibroblasts.

Lee Ju Won JW Yoo Hyuk Sang HS

ACS omega 20191101 20

To facilitate cell sheet formation of human dermal fibroblasts, gelatin moieties were chemically decorated onto the surface of electrospun nanofibrils (NFs). Poly(caprolactone) [PCL] was electrospun onto fibrous meshes and then fragmented into nanofibrils by optimized milling and hydrolysis. After aminolysis of the NFs, methacrylated gelatin (GelMA) was reacted via Michael-type addition with the surface-exposed amines of the aminolyzed NFs (ahPCL NFs). GelMA was immobilized on the ahPCL NFs. Ana ...[more]

PMID: 31737828

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Project description:Electrospinning is a simple and efficient method of fabricating a non-woven polymeric nanofiber matrix. However, using fluorinated alcohols as a solvent for the electrospinning of proteins often results in protein denaturation. TEM and circular dichroism analysis indicated a massive loss of triple-helical collagen from an electrospun collagen (EC) matrix, and the random coils were similar to those found in gelatin. Nevertheless, from mechanical testing we found the Young's modulus and ultimate tensile stresses of EC matrices were significantly higher than electrospun gelatin (EG) matrices because matrix stiffness can affect many cell behaviors such as cell adhesion, proliferation and differentiation. We hypothesize that the difference of matrix stiffness between EC and EG will affect intracellular signaling through the mechano-transducers Rho kinase (ROCK) and focal adhesion kinase (FAK) and subsequently regulates the osteogenic phenotype of MG63 osteoblast-like cells. From the results, we found there was no significant difference between the EC and EG matrices with respect to either cell attachment or proliferation rate. However, the gene expression levels of OPN, type I collagen, ALP, and OCN were significantly higher in MG63 osteoblast-like cells grown on the EC than in those grown on the EG. In addition, the phosphorylation levels of Y397-FAK, ERK1/2, BSP, and OPN proteins, as well as ALP activity, were also higher on the EC than on the EG. We further inhibited ROCK activation with Y27632 during differentiation to investigate its effects on matrix-mediated osteogenic differentiation. Results showed the extent of mineralization was decreased with inhibition after induction. Moreover, there is no significant difference between EC and EG. From the results of the protein levels of phosphorylated Y397-FAK, ERK1/2, BSP and OPN, ALP activity and mineral deposition, we speculate that the mechanism that influences the osteogenic differentiation of MG63 osteoblast-like cells on EC and EG is matrix stiffness and via ROCK-FAK-ERK1/2.

Dataset Information

Michael-Type Addition of Gelatin on Electrospun Nanofibrils for Self-Assembly of Cell Sheets Composed of Human Dermal Fibroblasts.

Publications

Michael-Type Addition of Gelatin on Electrospun Nanofibrils for Self-Assembly of Cell Sheets Composed of Human Dermal Fibroblasts.

Similar Datasets

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