Construction of 3-D Cellular Multi-Layers with Extracellular Matrix Assembly Using Magnetic Nanoparticles
- Authors
- Lee, Yu Bin; Lee, Joong-Yup; Ahmad, Taufiq; Bak, Seongwoo; Lee, Jinkyu; Kim, Hwan D.; Park, Tai Hyun; Hwang, Nathaniel S.; Lim, Youn Mook; Shin, Heungsoo
- Issue Date
- Oct-2016
- Publisher
- AMER SCIENTIFIC PUBLISHERS
- Keywords
- Hierarchical Tissue Mimicry; Multi-Layered Structure; 3-D Cellular Assembly; Magnetic Nanoparticles; Cellular Internalization
- Citation
- JOURNAL OF BIOMEDICAL NANOTECHNOLOGY, v.12, no.10, pp.1916 - 1928
- Indexed
- SCIE
SCOPUS
- Journal Title
- JOURNAL OF BIOMEDICAL NANOTECHNOLOGY
- Volume
- 12
- Number
- 10
- Start Page
- 1916
- End Page
- 1928
- URI
- https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/22130
- DOI
- 10.1166/jbn.2016.2303
- ISSN
- 1550-7033
- 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.
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