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Construction of 3-D Cellular Multi-Layers with Extracellular Matrix Assembly Using Magnetic Nanoparticles

Title
Construction of 3-D Cellular Multi-Layers with Extracellular Matrix Assembly Using Magnetic Nanoparticles
Author
신흥수
Keywords
Hierarchical Tissue Mimicry; Multi-Layered Structure; 3-D Cellular Assembly; Magnetic Nanoparticles; Cellular Internalization
Issue Date
2016-10
Publisher
AMER SCIENTIFIC PUBLISHERS
Citation
JOURNAL OF BIOMEDICAL NANOTECHNOLOGY, v. 12, NO 10, Page. 1916-1928
Abstract
Construction of 3-dimensional (3-D) engineered tissue is increasingly being investigated for use in drug discovery and regenerative medicine. Here, we developed multi-layered 3-D cellular assembly by using magnetic nanoparticles (MNP) isolated from Magnetospirillum sp. AMB-1 magnetotactic bacteria. Magnetized human dermal fibroblasts (HDFBs) were prepared by treatment with the MNP, induced to form 3-D assembly under a magnetic field. Analyses including LIVE/DEAD assay, transmission electron microscopy revealed that the MNP were internalized via clathrin-mediated endocytosis without cytotoxicity. The magnetized HDFBs could build 3-D structure as a function of seeding density. When the highest seeding density (5 x 10(5) cells/mm(2)) was used, the thickness of assembly was 41.0 +/- 1.69 mu m, with approximately 9.3 +/- 1.6 cell layers being formed. Immunofluorescence staining confirmed homogeneous distribution of ECM and junction proteins throughout the 3-D assembly. Real-time PCR analysis showed decrease in expression levels of collagen types I and IV but increase in that of connexin 43 in the 3-D assembly compared with the 2-D culture. Finally, we demonstrated that the discernible layers can be formed hierarchically by serial assembly. In conclusion, our study showed that a multi-layered structure can be easily prepared using magnetically-assisted cellular assembly with highlighting cell-cell and cell-ECM communication.
URI
https://www.ingentaconnect.com/content/asp/jbn/2016/00000012/00000010/art00007https://repository.hanyang.ac.kr/handle/20.500.11754/100516
ISSN
1550-7033; 1550-7041
DOI
10.1166/jbn.2016.2303
Appears in Collections:
COLLEGE OF ENGINEERING[S](공과대학) > BIOENGINEERING(생명공학과) > Articles
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