The effect of growth factors and extracellular matrices on periodontal ligament cell differentiation during periodontal tissue regeneration.

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The roles of growth factors and extracellular matrices were investigated for the improvement of periodntal regeneration therapy. 1., We first forcused this study on basic fibroblast growth factor (bFGF) and established the anti-human bFGF monoclonal antibodies (mAbs). One of those Abs, BF-24 was utilyzed for immunohistochemical analysisand showed the dense distribution of bFGF especially in periodontal ligament (PDL) and epithelium. In order to examine the involvement of bFGF during the tissue regeneration ana wound healing, artificial three wall intrabony defect was prepared in beagle dogs. BF-24 was able to detect the prominent expression of bFGF one week after surgery transiently. 2, Mouse monoclonal PDL cell, MPDL-22 was established in the presence of bFGF.MPDL-22 revealed long term proliferation in the presence of bFGF,however, once bFGF was deprived, it lead the to maturation and started calcification. The mRNA of bone matix protein, such as osteocalcin, osteopontin, and cementum protein, type III collagen and ligamentum protein, typeXII collagen was increased after the maturation of MPDL-22, suggesting that the expression of these protein may be a differentiation marker for perodontal ligament tissue. 3, In the case of human HPDL,the stimulation by bFGF increased the expression of laminin mRNA.In contrast, the amount of type I collagen mRNA was decreased and no significant alteration was observed in fibronectin and type II collagen mRNA expression. 4, The acid extract of cementum was fractionated in order to identify unknown ligament growth factor. Several step of biochemical purification resulted that m.w.10-20kDa protein contained the mitogenic activity for HPDL and the sequence analysis of peptide fragment revealed its partial homology to Insulin like growth factor-I.