Studies on differentiation and proliferation factors for induction of biological reparative dentin formation

诱导生物修复牙本质形成的分化和增殖因子的研究

基本信息

批准号：
09671951
负责人：
TAKASHIBA Shogo
金额：
$ 2.24万
依托单位：
Okayama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

项目摘要

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 preparationTo 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 tubulesFreshly 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) fibroblastsBy 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 strainsGenes of Porphyromonas gingivalis strains were compared at genomic DNA level by SH.This result was used to apply this method for eukaryotic cells.

为了有效诱导继发性牙本质，需要允许牙本牙髓中的次生牙本质诱导因子表达。这些因素必须充当特定于牙髓细胞的分化和增殖因子，并且必须阐明其特征和功能，在这项研究中，我们成功地跟随了：1）建立了将独特基因与组织和细胞分离的方法（真核生物和原核子）（2）筛查牙科造成的牙齿pulp中的筛查基因筛查。 3）通过牙本质管将污渍转移到纸浆腔室。特定点的实验结果1。腔制剂分离以分离这些基因后，在牙髓中独特地表达的基因，我们需要精确的方法来检测基因表达从少量组织中的变化。基于聚合酶链反应（PCR）的减法杂交（SI-I）方法使我们能够分离出腔制剂后在牙髓中独特表达的cDNA.2。将基因或其产物转移到纸浆腔室的模型，该模型使用含有重要的牙髓组织的牙本质小管萃取的人类牙齿的模型来建立该模型。腔制备后几天，准备牙齿以进行器官培养。培养开始时，将污渍放入腔体中，并在几天后发现在纸浆腔中。该结果表明，基因通过牙本质小管转移到纸浆组织的可能性3。由人类牙周韧带（PDL）成纤维细胞与PDL成纤维细胞和牙龈成纤维细胞之间减去杂交（两者都是原代培养物）在唯一表达的基因，将几种基因阐明以通过PDL成纤维细胞独特地表达。该实验使我们能够改善Sh.4。通过Sh，在基因组DNA水平上比较了牙龈卟啉单胞菌菌株的卟啉单胞菌之间的基因差异。该结果用于将此方法应用于真核细胞。