IADR Abstract Archives
Construction Of Tooth Germ Cell Sheet Applying Magnetic Nanoparticles
Objectives: Regenerating dental enamel still remains difficult due to its complicated developmental process. It has been reported that it is essential to control the reciprocal interaction between dental epithelial cells (DEC) and dental mesenchymal cells (DMC), as in the development of tooth germ. Therefore, we sought to control the cell arrangement by using magnetic nanoparticles (MNP). In this research, we combined the layers of DEC and DMC to construct a combination cell sheet (DEC/DMC sheet) using the magnetic tissue engineering system (Mag-TE) and investigated the expressions of dental specific markers in the cell sheet.
Methods: DEC and DMC were isolated from third molar tooth bud of 6-month-old porcine lower jaw. The DEC/DMC sheet was constructed using the Mag-TE in one dish. The expressions of the differentiation markers and basement membrane markers including amelogenin, enamelin, ameloblastin, RUNX2, collagen type I alpha 2 (COL1α2), dentin sialophosphoprotein, and collagen type IV alpha 1 (COL4α1) in the DEC/DMC sheet were examined by real-time RT-PCR and immunofluorescent staining. Mixtures of a DEC sheet and a DMC sheet made in different dishes, respectively, were used as the controls (Student t-test, p<0.05).
Results: The mRNA expressions of amelogenin, enamelin, ameloblastin, RUNX2, and COL4α1 in the DEC/DMC sheet increased as compared with those in the control (p<0.05). Immunostaining showed COL4α1 expressed at the border region between the DEC and DMC layers in the DEC/DMC sheet.
Conclusions: In vitro studies revealed that the epithelial–mesenchymal intereaction between DEC and DMC enhanced by magnetic force to bind DEC and DMC. It suggests that the microenvironment in the DEC/DMC sheet might be similar to that in the developmental stage of a tooth bud. Therefore, DEC/DMC sheets constructed using a Mag-TE could be developed into a ideal graft for artificially regenerating dental enamel.
Methods: DEC and DMC were isolated from third molar tooth bud of 6-month-old porcine lower jaw. The DEC/DMC sheet was constructed using the Mag-TE in one dish. The expressions of the differentiation markers and basement membrane markers including amelogenin, enamelin, ameloblastin, RUNX2, collagen type I alpha 2 (COL1α2), dentin sialophosphoprotein, and collagen type IV alpha 1 (COL4α1) in the DEC/DMC sheet were examined by real-time RT-PCR and immunofluorescent staining. Mixtures of a DEC sheet and a DMC sheet made in different dishes, respectively, were used as the controls (Student t-test, p<0.05).
Results: The mRNA expressions of amelogenin, enamelin, ameloblastin, RUNX2, and COL4α1 in the DEC/DMC sheet increased as compared with those in the control (p<0.05). Immunostaining showed COL4α1 expressed at the border region between the DEC and DMC layers in the DEC/DMC sheet.
Conclusions: In vitro studies revealed that the epithelial–mesenchymal intereaction between DEC and DMC enhanced by magnetic force to bind DEC and DMC. It suggests that the microenvironment in the DEC/DMC sheet might be similar to that in the developmental stage of a tooth bud. Therefore, DEC/DMC sheets constructed using a Mag-TE could be developed into a ideal graft for artificially regenerating dental enamel.