|Budget Amount *help
¥7,300,000 (Direct Cost : ¥7,300,000)
Fiscal Year 1997 : ¥1,100,000 (Direct Cost : ¥1,100,000)
Fiscal Year 1996 : ¥6,200,000 (Direct Cost : ¥6,200,000)
[Purpose] Our final aim is to develop the new direct pulp capping agents (for example, tetracalcium phosphate (4CP) and tetracalcium phosphate-citric acid complex cement (4CPC).
We already reported that 4CP and 4CPC showed their effects on the dog pulp cell through extracellular matrixes. Although the medium of the pulp cells usually contains fetal bovine serum, this serum has many extracellular matrixes.
The purposes of this research were to establish the serum free culture system of the dog pulp cells and to check the effects of extracellular matrixes on these dog pulp cells.
[Result] The condition of serum free culture medium of the dog pulp cell was alpha MEM containing transferrin (20 mug/ml), bFGF (1 ng/ml), egg lipoprotein (50 mug/ml), hydrocortisone (10^<-9>-10^<-8>), insulin (5 mug/ml) and EGF (1 ng/ml). With this culture condition, the dog pulp cells could grow and produce alkaline phosphatase (ALPase) as well as with 20% serum medium. This serum free cultured pulp cells mainly produced type I collagen. These results showed that we could culture the dog pulp cells with this serum free medium.
Next, we checked the effects of many extracellular matrixes (type I collagen, type III collagen, type IV collagen, plasma fibronectin, cellular fibronectin and laminin) on dog pulp cells that were cultured with this serum free medium. The effects of these extracellular matrixes were almost same. These extracellular matrixes promoted the attachment of the pulp cells to the dishes and growth of the cells and production of ALPase.
[Conclusion] We established the serum free culture condition of the dog pulp cells. Under this condition, the extracellular matrixes could promote the attachment of the pulp cells to the dishes and the growth of the cells and the production of ALPase. These results show that we can check the effects of 4CP and 4CPC on these pulp cells with this serum free medium and develop the new direct pulp capping agents.