The Expression Of KGF In The Injury Process Of Inflammation Of Human Gingival Fibroblasts

Objectives: (1) The human gingival fibroblasts (HGFs) will be cultured in vitro and its model will be constructed in order to provide the experimental conditions for the further research.(2) To observe the changes of gene expression level in the inflammation injury of keratinocyte growth factor (KGF)with the intervention of IL-1βand IL-6 under different concentrations, and that is for further confirmation of the possible function of KGF in the occurrence and development of periodontal disease.Methods: (1) The freeze-stored human gingival fibroblasts (HGFs) in liquid nitrogen will be thawed, passaged and cultured, and then it will be tested by morphology and immunohistochemistry. To observe the properties of the cells after cryopreservation and anabiosis so as to establish a stable model. (2) Choose the fifth to eighth human gingival fibroblasts (HGFs) which are stable in the process of growth and are morphologically good. Moreover, use IL-1βof 0ng/ml,0.1ng/ml,0.5ng/ml,1ng/ml,5ng/ml,10ng/ml and IL-6 of 0ng/ml,0.1ng/ml,0.5ng/ml,1ng/ml,5ng/ml,10ng/ml on cells intervention respectively,then establish the cells model of the inflammation injury. Using reverse transcription-polymerase chain reaction (RT-PCR) to detect the expression of KGF genes in each experimental group,and collecting the test results including images and datum by computer image analysis system.As a consequence, the expression of KGF genes in the human gingival fibroblasts (HGFs) under different concentrations of IL-βand IL-6 intervention will be statistically analysed by means of single-factor variance analysis.Results: (1) Having been thawed,passaged and cultured,the freeze-stored fifth to eighth HGFs in liquid nitrogen were stable in the process of growth and were morphologically good.The cells presented either long spindle or star appearance which is typically fibroblast-like cell characteristics under invert microscope.The antibodies against ceratin with immunohistochemis- try analysis.(2) RT-PCR results showed that in terms of gene level,the KGF in normal human gingival fibroblasts had a weak-positive quality while the expression of KGF showed the gradual increasing trend with the concentrations of IL-βand IL-6 increasing from 0.1ng/ml to 10ng/ml. Conclusions: (1) The thawed,passaged and cultured human gingival fibroblasts accord with the characteristics of the morphology and immunology with cells growing stably and showing vigorously good.Accordingly,the cells can be used to be cells model to provide experimental basis for the following research.(2) As for the gene level research,this experiment also shows that KGF has a relation to the inflammation process of human gingival fibroblasts which IL-βand IL-6 cause,which is likely to play an important role in the occurrence and development of periodontal disease,thus providng a new theoretical basis for further research of the pathogenesis of periodontal disease.