| Background and objective Chronic periodontitis is a chronic inflammatory disease caused by the interaction of periodontal pathogens and host immune defense reaction.Its clinical manifestations are gingival inflammation,loss of clinical attachment and alveolar bone resorption.Type 2 diabetes mellitus(T2DM)is a kind of chronic metabolic disorder disease,which is mainly manifested by the increase of blood glucose.The body is in a high glucose state for a long time,leading to various system diseases.Diabetes is the main risk factor of periodontal disease.In diabetic patients with periodontitis,continuous hyperglycemia results in nonenzymatic glycosylation of various proteins in the body.The advanced glycation end products(AGEs)formed by them play an important role in the pathogenesis of diabetic periodontitis.Recently,it has been found that macrophages(M_φ),as an important part of the innate immune system,can play a defensive role in periodontal tissues and maintain periodontal tissue homeostasis.In addition,they can also act on the body through the release of pro-inflammatory factors,leading to periodontal tissue damage and alveolar bone absorption,which is an essential immune cell component in the pathogenesis of periodontitis.Studies have shown that M_φ can be detected in normal gingival tissue and periodontitis gingival tissue.Calprotectin(S100A8/A9 complex),also known as calprotectin,is mainly secreted by monocytes,neutrophils and M_φ.It is a heterodimer S100A8/A9 protein complex formed by S100A8(MRP-8)and S100A9(MRP-14)in a calcium dependent manner.It is found that calprotectin is closely related to many inflammatory diseases,autoimmune diseases,T2 DM and periodontitis.In physiological condition,calprotectin is expressed in gingival crevicular fluid,serum and saliva of normal body.In patients with periodontitis,especially in gingival tissue and serum of diabetic patients with periodontitis,the expression of S100A8/A9 is higher than that of gingival tissue and serum of periodontal healthy people.However,the expression mechanism of S100A8 and S100A9 in T2 DM is not clear,and there is no detailed report about the expression of S100A8/A9 on macrophages in gingival tissue of diabetes mellitus with periodontitis.In this study,the expression of S100A8/A9 on macrophages in gingival tissue of diabetes mellitus with periodontitis was observed to explore the role and mechanism of S100A8 and S100A9 on macrophages in diabetic patients with periodontitis.The results of this study will provide the molecular theoretical basis for the follow-up immune prevention and treatment of diabetes mellitus with periodontitis targeted at macrophages S100A8 and S100A9.Methods Ninety-four subjects who voluntarily participated in the study were divided into 5 groups according to the classification criteria: 20 in the normal control group,20 in the slight chronic periodontitis group,20 in the moderate periodontitis group,20 in the severe chronic periodontitis group,and 14 in the type 2 diabetes mellitus with periodontitis group.The specimens were fixed in 4 % paraformaldehyde solution for more than 48 hours and made into 5μm continuous sections.The expression of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ in gingival tissue was observed by double immunofluorescence staining(DIF).Results 1.Histological observations 1)Compared with the normal control group,the scores of inflammation degree in the four chronic periodontitis groups increased significantly(P < 0.01);2)In the chronic periodontitis group,the scores of inflammation degree increased with the severity of periodontitis significantly(P < 0.01);3)There was no significant difference in scores of inflammation degree between type 2 diabetes mellitus with periodontitis group and severe chronic periodontitis group(P > 0.05).2.Double-immunofluorescence staining results 1)Compared with the normal control group,the density of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ in gingival tissue of the four groups of patients with periodontitis significantly increased(P < 0.01);2)The density of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ in the moderate chronic periodontitis group was significantly higher than that in the slight chronic periodontitis group(P < 0.01);3)The density of CD68-S100A8 and CD68-S100A9 double positive M_φ in severe chronic periodontitis group was significantly higher than that in slight and moderate chronic periodontitis group(P < 0.01);4)The density of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ in type 2 diabetic with periodontitis group was significantly higher than that in slight,moderate and severe chronic periodontitis group(P < 0.01);3.Correlation analysis results were obtained between the degree of chronic periodontitis and CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ By Pearson correlation analysis,the density of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ was positively correlated with the severity degree of periodontitis.Conclusions The gingival tissues of 94 patients with chronic periodontitis and type 2 diabetes mellitus with periodontitis in different pathological degrees were observed.The expression of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ in gingival tissues was observed by immunofluorescence double staining.The conclusions are as follows: 1.In chronic periodontitis,with the aggravation of the pathological changes of periodontitis,the score of inflammation degree increased significantly.However,there was no significant difference in the score of inflammation degree between type 2 diabetes mellitus with periodontitis and severe chronic periodontitis.2.The cell density of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ increased with the severity of periodontitis.3.The density of CD68-S100A8 double positive M_φ and CD68-S100A9 double positive M_φ in type 2 diabetes with periodontitis group was significantly higher than that in slight,moderate and severe chronic periodontitis group. |
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