Immortalized Gingival Fibroblasts for Cytotoxicity Test Model of Dental Materials
Objectives: This study was designed to evaluate whether hTERT-transfected immortalized human gingival fibroblasts (hTERT-F) are more suitable to test cytotoxicity of dental materials, compared to L929 malignant cells derived from mouse fibroblasts.
Methods: The immortalization of normal human gingival fibroblast cell line (NOF) was established by transfection of retroviral vector plpc-hTERT. hTERT transfection was confirmed by western blotting and TRAP assay. Biological properties such as proliferative activity, migration activity, cell morphology, cell cycle status, and collagen production were compared among NOF, hTERT-F and L929. Cytotoxicity tests of dental materials were performed using these three cell lines. Results: hTERT transfection was confirmed by higher hTERT expression and increased telomerase activity in hTERT-F, compared to NOF.Proliferation rate of hTERT-F at the early passages was lower than NOF at the similar stages. However, NOF over 28th passage showed growth retardation, while hTERT–F keep on proliferating over 30th passage, showing higher proliferation rate along increased passages. In contrast, L929 has the highest proliferation rate among three cell lines, showing different cell cycle pattern compared to NOF and hTERT-F. Immunofluorescence showed that NOF and hTERT-F harbor both vimentin and actin, while L929 showed no detectable vimentin expression. However, migration activity and collagen production showed no significant difference among three cell lines. Cytotoxicity test of dental materials showed similar patterns among three cell lines.
Conclusion: hTERT-F is successfully immortalized and has similar biological characteristics to NOF. L929 has markedly deviated biological properties compared to NOF. These results suggest that hTERT-F would be a suitable candidate for cytotoxicity test of dental materials.
Acknowledgment: Supported by PRC Programme (2009-0094028) and NRF (R13-2003-013-04001-0) funded by the government of Korea.
Division: IADR/PER General Session
Meeting:2010 IADR/PER General Session (Barcelona, Spain) Location: Barcelona, Spain
Year: 2010 Final Presentation ID:540 Abstract Category|Abstract Category(s):Dental Materials 5: Biocompatibility and Biologic Effects
Authors
Illeperuma, Rasika Pawiththra
( Oral Cancer Research Institute, Department of Oral Pathology, Yonsei University College of Dentistry, Seoul, N/A, South Korea
)
Park, Young Jin
( Oral Cancer Research Institute, Department of Oral Pathology, Yonsei University College of Dentistry, Seoul, N/A, South Korea
)
Kim, Jin Mi
( Oral Cancer Research Institute, Department of Oral Pathology, Yonsei University College of Dentistry, Seoul, N/A, South Korea
)
Che, Zhong Min
( Oral Cancer Research Institute, Department of Oral Pathology, Yonsei University College of Dentistry, Seoul, N/A, South Korea
)
Han, Mi Ra
( Deptartment of Dental Biomaterials & Bioengineering,Yonsei University College of Dentistry, Seoul, N/A, South Korea
)
Kim, Kwang Mahn
( Deptartment of Dental Biomaterials & Bioengineering,Yonsei University College of Dentistry, Seoul, N/A, South Korea
)
Kim, Jin
( Oral Cancer Research Institute, Department of Oral Pathology, Yonsei University College of Dentistry, Seoul, N/A, South Korea
)
SESSION INFORMATION
Poster Discussion Session
Biocompatibility and Biologic Effects (PDS) I
07/15/2010