1998 Fiscal Year Final Research Report Summary
Studies on differentiation and proliferation factors for induction of biological reparative dentin formation
| Project/Area Number |
09671951
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 | Okayama University |
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Principal Investigator |
TAKASHIBA Shogo Okayama Univ., Dental School, Associate Professor, 歯学部, 助教授 (50226768)
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| Co-Investigator(Kenkyū-buntansha) |
WASHIO Norifumi Okayama Univ., Dental School, Instructor, 歯学部, 助手 (40263602)
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| Project Period (FY) |
1997 – 1998
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| Keywords | Dental Pulp / Calcification / Secondary dentin / Gene expression / Subtractive hybridization |
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| Research Abstract |
For the effective induction of secondary dentin, it is needed to allow the secondary dentin-inducible factors express in dental pulp. These factors must act as differentiation and proliferation factors specific for dental pulp cells, and have to be elucidated their characters and functions, In this study, we succeeded followings : 1) establishment of method to isolate unique genes from tissue and cells (eukaryote and prokaryote), 2) screening of cavity preparation-induced gene in dental pulp. and 3) transfer of stains to pulp chamber through dentinal tubes.
Experimental results for the specific points.
1. Genes expressed uniquely in dental pulp after cavity preparation
To isolate these genes, we needed precise method to detect change of gene expression from small amount of tissue. Polymerase chain reaction (PCR)-based subtractive hybridization (SI-I) method allowed us to isolate cDNAs uniquely expressed in dental pulp after cavity preparation.
2. Model for the transfer of genes or their products to pulp chamber through dentinal tubules
Freshly extracted human teeth which contain vital pulp tissue were used for establishment of this model. The teeth were prepared for organ culture for several days after cavity preparation. Stains were placed into the cavity at the beginning of culture, and found to be in pulp chamber after a couple of days. This result suggests that the possibility of gene transfer to pulp tissue through dentinal tubules.
3. Genes expressed uniquely by human periodontal ligament (PDL) fibroblasts
By subtractive hybridization between PDL fibroblasts and gingival fibroblasts (both are primary culture), several genes were elucidated to be expressed uniquely by PDL fibroblasts. This experiment allowed us to improve SH.
4. Difference of genes between Porphyromonas gingivalis strains
Genes of Porphyromonas gingivalis strains were compared at genomic DNA level by SH.This result was used to apply this method for eukaryotic cells.
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