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The Effect of Basic Fibroblast Growth Factor on the Expression of Adisinteghn and Metall-proteinase with Thrombospondin Motif-1by Human Periodontal Ligament Cells

Author: QinYanYan
Tutor: LinChongZuo
School: Jilin University
Course: Clinical Stomatology
Keywords: A disintegrin and metalloproteinase with thrombospondin motif-1 hPDLC bFGF syndecan-4
CLC: R781.4
Type: Master's thesis
Year: 2012
Downloads: 29
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Abstract


Periodontitics is a chronic and infectional disease which is caused bybiofilm in dental plaque. It leads to the supporting tissues destruction such aspockets, alveolar bone defects and attachment loss, even the teeth get loose andlost. Investigators have reexamined periodontal infections can have an adverseeffect on general health and that systemic diseases can accelerate thedevelopment of periodontal diseases.The main goal of periodontal treatment isto remove dental plaque,control inflammation,especially gain tissue repair andregeneration. But the gainning tissue regeneration is limited with presentperiodontal treatment. Recent years,with the development of tissue engineeringwhich is formed by cells,growth factors and scaffold,people put their hope inPeriodontal tissue engineering. Only periodontal ligament cells which is theideal seed cells can be used as stem cells in wound healing of periodontal tissue.The adhesion,proliferation,and differentiation of periodontal ligament cells onthe root surface are the key component of gaining periodontal tissueregeneration.Fibroblast growth factor2(FGF-2) is polypeptide molecule whichmediates a variety of physiological and pathological responses in developmentand wound healing.In vitro studies reveals that FGF-2can promote periodontalligament cells proliferation, migration and adhesion. However the detailedmechanism by that FGF-2regulate periodontal ligament cells remains elusive.Extracellular matrix synthesis and degradation are involved in wound heal.The syndecan can modulate cell proliferation and cell migration and regulatesignaling communication between cell and extracellulur matrix during woundreparation. As co-receptors of FGF, syndecans increase the local concentrationsof growth factors and promote signaling cascades. FGF-2regulate signaling communication, promote cell proliferation and migration by syndecan-4on cellsurface.Protease of ECM plays a key role in wound healing.A disintegrin andmetalloproteinase with thrombospondin motifs-1(ADAMTS-1) is a member ofthe family of multifunctional proteins known as ADAMTS. Several studieshave shown that ADAMTS-1is capable of degrading aggrecan andversican,which are components of extracellular matrix (ECM) barriers,andplays a key role in many physiology and pathology processes. In skin woundheal, low-level ADAMTS1promotes endothelial and fibroblast cells migrationby degrading ECM protein, but high-level ADAMTS1can combine withFGF-2to inhibite endothelial and fibroblast cells migration.ADAMTS familymembers are syndecan-4sheddases.ADAMTS1uniquely cleave thetransmembrane proteoglycan syndecan-4, and the extracellular domainshedding made cell skeletal structure change which help cell to migration. Thecurrent findings show that syndecan-4increases obviously in conditionedmedium of FGF-2-activated HPDLCs and reduces on the HPDLCs surface.It’snot clear that the role of ADAMTS1in periodontal tissue wound heal at homeand abroad.As mentioned above, our study choose human periodontal ligament cellscultured in vitro using tissue explant as subject to investigate the expression ofADAMTS1of PDLCs and the effect of bFGF on ADAMTS1gene expression,and to explore the role of ADAMTS1in periodontal tissue wound heal. Itprovides a new view for the mechanism that how bFGF promote PDLCsproliferation and migration.Methods: We gathered healthy premolars of young people(10-15years old)undergoing tooth extraction for orthodontic treatment in stomatology of JilinUniversity,washed tooth with stroke-physiological saline solution, removeblood and bacteria at surface of the root. Healthy periodontal tissue was removed from the center of the root surface with a surgical scalpel andsubgingival curette. We used tissue explant to cultured human periodontalligament cells in vitro. Experiments were carried out with cells from the third tofifth passages. Immunohisto-chemical method was used to examine theexression of ADAMTS1in PDLCs. Expression of ADAMTS1mRNA ofPDLCs stimulated by bFGF was analyzed by reverse transcriptase-Polylnerasechain reaction on24h,48h,72h.Result: we found ADAMTS1is positive expression in PDLCs.10ng/mlbFGF significantly stimulate ADAMTS1expression in PDLCs compared withthe control. ADAMTS1was expressed at every time point, and the level ofADAMTS1at48h is higher comparing with24h(P<0.01),72h(P<0.01), and24h is higher than72h(P<0.01).Conclusion: ADAMTS1is positive expression in PDLCs.10ng/ml bFGFsignificantly stimulate ADAMTS1expression of PDLCs at the earlier period ofproliferation.

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