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TISSUE-SPECIFIC STEM CELLS |
aDepartment of Orthopaedics and Traumatology, Taipei Veterans General Hospital, Taipei, Taiwan;
bInstitute of Biopharmaceutical Sciences, National Yang-Ming University, Taipei, Taiwan;
cBiomedical Engineering Research Laboratory, Industrial Technology Research Institute, Hsinchu, Taiwan;
dInstitute of Biochemical and Biomedical Engineering, Chang-Gung University, Taoyuan, Taiwan;
eInstitute of Biomedical Engineering, National Yang-Ming University, Taipei, Taiwan;
fDepartment of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan;
gSchool of Medicine, National Yang-Ming University, Taipei, Taiwan
Key Words. Mesenchymal stem cells • Electrospin • Nanofibers • Type I collagen • Focal adhesion
Correspondence: Oscar K. Lee, M.D., Ph.D., 201 Shi-Pai Road, Sec. 2, Taipei 11221, Taiwan. Telephone: +886-2-2875-7557; Fax: +886-2-2875-7657; e-mail: kslee{at}vghtpe.gov.tw
Received April 25, 2006;
accepted for publication June 23, 2006.
We reconstituted type I collagen nanofibers prepared by electrospin technology and examined the morphology, growth, adhesion, cell motility, and osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (MSCs) on three nano-sized diameters (50–200, 200–500, and 500–1,000 nm). Results from scanning electron microscopy showed that cells on the nanofibers had a more polygonal and flattened cell morphology. MTS (3-[4,5-dimethythiazol-2-yl]-5-[3-carboxy-methoxyphenyl]-2-[4-sul-fophenyl]-2H-tetrazolium compound) assay demonstrated that the MSCs grown on 500–1,000-nm nanofibers had significantly higher cell viability than the tissue culture polystyrene control. A decreased amount of focal adhesion formation was apparent in which quantifiable staining area of the cytoplasmic protein vinculin for the 200–500-nm nanofibers was 39% less compared with control, whereas the area of quantifiable vinculin staining was 45% less for both the 200–500-nm and 500–1,000-nm nanofibers. The distances of cell migration were quantified on green fluorescent protein-nucleofected cells and was 56.7%, 37.3%, and 46.3% for 50–200, 200–500, and 500–1,000 nm, respectively, compared with those on the control. Alkaline phosphatase activity demonstrated no differences after 12 days of osteogenic differentiation, and reverse transcription-polymerase chain reaction (RT-PCR) analysis showed comparable osteogenic gene expression of osteocalcin, osteonectin, and ostepontin between cells differentiated on polystyrene and nanofiber surfaces. Moreover, single-cell RT-PCR of type I collagen gene expression demonstrated higher expression on cells seeded on the nanofibers. Therefore, type I collagen nanofibers support the growth of MSCs without compromising their osteogenic differentiation capability and can be used as a scaffold for bone tissue engineering to facilitate intramembranous bone formation. Further efforts are necessary to enhance their biomimetic properties.
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