2006 Fiscal Year Final Research Report Summary
DEVELOPMENT OF DENTINE REGENERATION THERAPY BY GENE DELIVARY SYSTEM USING NATURAL BIOACTIVE MATERIAL, CHITOSAN
| Project/Area Number |
16390547
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Conservative dentistry
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| Research Institution | Nagasaki University |
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Principal Investigator |
HAYASHI Yoshihiko Nagasaki University, GRADUATE SCHOOL OF BIOMEDICAL SCIENCES, PROFESSOR, 大学院医歯薬学総合研究科, 教授 (20150477)
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| Co-Investigator(Kenkyū-buntansha) |
YANAGIGUCHI Kajiro Nagasaki University, UNIVERSITY HOSPITAL, LECTURER, 医学部・歯学部附属病院, 講師 (50264255)
OHARA Naoko Nagasaki University, GRADUATE SCHOOL OF BIOMEDICAL SCIENCES, ASSISTANT PROFESSOR, 大学院医歯薬学総合研究科, 助手 (80301365)
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| Project Period (FY) |
2004 – 2006
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| Keywords | CHITOSAN / CELL ACTIVATION / SIGNAL TRANSDUCTION / INTRODUCTION OF GENE / CELL DIFFERENTIATION・PROLIFERATION / ELECTROPORATION |
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| Research Abstract |
Chitosan has many bioactive effects. We discovered that chitosan monomer promotes the acceleration of alkaline phosphatase activity in osteablastic NOS-1 cells through BMP-2 expression. After that, we tried to demonstrate whether chitosan monomer activated pathways of signal transduction in NOS-1 cells using cDNA microarray and real-time PCR analyses.
cDNA microarray analysis revealed that 10 genes conserning to various signaling-related molecules were expressed at 【greater than or equal】 2.0-fold higher ratio levels in the experimental group when compared with the control group after 3 days. Real-time PCR analysis showed that chitosan monomer induced an increase in the expression of four signal transduction genes. Furthrmore, MAPKs antibo lies array indicated that these molecules were phosphorylated. We concluded that a super-low cincentration of chitosan monomer could modulate the activity of osteoblastic cells through mRNA levels and that chitosan monomer directly affects signal transduction pathways.
The aforementioned data conduct the application of chitosan to the induction of gene, which means the protection effect of chitosan during electroporation of DNA into the cells. We investigated the induction of BMP-2 gene into NOS-1 cells using electroporation. The marker of successful induction was judged with the fluorescence of GFP inside the cells. The supplementation of chitosan monomer during the electroporation produced that the proliferation of the induced cells were much higher in the chitosan group. These findings suggest that the application of chitosan during electroporation would be useful for maintaining cell vitality in pulp wound healing through dentine regeneration in vivo.
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