| Objective To detect the change of Cathepsin k concentration in gingival crevicular fluid(GCF)in healthy subjects and patients with different degrees of periodontitis,and to explore whether there is a correlation between the inflammatory degree of periodontitis and the expression level of GCF,whether it can be used as a reference index to reflect the degree of periodontitis damage and provide a theoretical basis for assisting the diagnosis of periodontitis.Methods1.Objectives From October 9,2018 to September 14,2019,104 patients from the Department of Stomatology,Affiliated Hospital of Chifeng University were selected as the control group(group A),and the patients with periodontal health were required to have a depth of exploration ≤ 3mm,no attachment loss and alveolar bone absorption,26 in total,13 men and 13 women,with an average age of 38.12 years;the patients diagnosed as periodontitis were set as group B,according to the new international classification standard of periodontitis,the first three stages of periodontitis were divided into threegroups: stage I periodontitis(Group B1),stage II periodontitis(Group B2)and stage III periodontitis(Group B3).Each group was also 26 people,a total of 78 people,37 men and 41 women,with an average age of 42.50 years.The selected patients had informed consent to the purpose and process of the trial,and voluntarily participated in the trial.The selected patients were required to have no systemic diseases,no antibiotics and non steroidal antiinflammatory drugs within 3 months,no periodontal treatment within 6 months,no smoking,and no pregnancy.2.Data acquisition Two groups of patients were measured the related indexes of mandibular molars,including gingival index(GI),probing depth(PD),loss of attachment(AL),X-ray showing alveolar bone absorption(ABL)3.Gingival crevicular fluid collection Remove the tip of No.30 hygroscopic paper,remove the 4.5 mm part of the tip,and sterilize it by irradiation.Four paper points for a group,into the EP pipe has been completed disinfection,number,weighing,after recording for use.The gingival crevicular fluid was collected by inserting the hygroscopic paper tip into the gingival pocket at the selected collection point.After collection,4 moisture-absorbing paper tips were placed in the EP tube and weighed again.The number and weight were recorded.The weight difference between the front and back of the EP tube was the weight of GCF collected.After recording,seal the EP tube and place it in the refrigerator at-70 °C for inspection.Defrost the sample at room temperature,add buffer,shock,centrifugation,take part of the supernatant,to be tested.4.The concentration of Cathepsin K was detected by Elisa A blank hole,a standard hole and a sample hole are arranged on the enzyme label plate.Add the sample in the hole of standard sample,add thediluted sample solution in the hole of sample to be tested,close the plate and warm it up and then shake it dry.The enzyme labelled reagent(except the blank hole)is added into the holes after washing,and the sealing plate is warmed and then shaken dry.Then the developer A and developer B are successively added into the hole after washing,shake gently and mix well,avoid light for 15 minutes.The termination solution is then added and the OD value of each hole is immediately measured.SPSS25.0 statistical software for statistical analysis,selection of appropriate statistical methods for testing.Results The average concentration of Cathepsin K in gingival crevicular fluid of periodontal healthy control group(group A)was 53.26±5.70pg/ml;the average value of Cathepsin K in gingival crevicular fluid of periodontitis experimental group B1 was 67.66±4.40pg/ml,the average value of group B2 was 64.72±6.04pg/ml,and the average value of group B3 was 61.27±4.67pg/ml.The concentration of Cathepsin K in the periodontal healthy control group was lower than that in the periodontitis experimental groups(P<0.05).As the periodontal inflammation gradually increased,the concentration of Cathepsin K in the three groups B1,B2,and B3 gradually decreased,and there were differences between the four groups(P<0.05).The concentration of Cathepsin K in the experimental group was not significantly correlated with GI(P>0.05),but negatively correlated with PD,AL,and ABL(P<0.01).The difference was statistically significant.Conclusions 1.The concentration of Cathepsin K in gingival crevicular fluid in periodontal healthy control group was lower than that in periodontitis experimental group.With the increase of periodontal inflammation,the concentration of Cathepsin K in gingival crevicular fluid decreased gradually.2.The inflammatory degree of periodontitis was correlated with the expression level of Cathepsin K in gingival crevicular fluid,which might be a reference index to reflect the severity of periodontitis. |
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