Correlation Between Biomarkers And Peri-implantitis

In clinic,7.8%-43.3% of implant dentures have failed,and peri-implantitis represents one of the important causes of the implant failure.The commonly used clinical examinations,such as the probing pocket depth(PPD),gingival bleeding index(GBI),and X-ray,can only indicate the disease severity,but cannot predict and judge the entire disease process and development.It has been shown that some biomarkers and their content changes in patients with peri-implantitis are closely related to the disease occurrence and development.Monitoring and detecting of these biomarkers might contribute to the evaluate and predict the disease activity.However,the evaluation based on single biomarker or index would be incomplete,while the marker packages(combining various indicators)might provide more accurate and comprehensive prediction,diagnosis,and evaluation of the disease.In this study,the implant gingival crevicular fluid was used as sample,and four major types of biomarkers related to peri-implantitis were selected based on the marker packages,i.e.,the proteomic biomarkers,genetic biomarkers,microbial biomarkers,and other biomarkers,including the following representative indicators with certain sensitivities: IL-1?,IL-17,RANKL,MMP-8,MMP-13,CRP,IL-1a and subgingival flora.The contents of multiple cytokines were simultaneously detected with Procartaplex multi-cytokine detection kits based on the Luminex platform.The bacterial DNA was extracted from the samples,and the 16 SrRNA V4-V5 regions were sequenced on the Illumina Miseq sequencing platform and Illumina PE250 sequencing platform.The subgingival flora were evaluated according to the Phylum,Class,Order,Family,and Genus.This study aimed to use advanced detection methods to investigate the differences in the biomarker levels in gingival crevicular fluid between healthy implant and peri-implantitis,the characteristics of the subgingival flora in Uygur patients with peri-implantitis,and the difference in the expression of biomarkers in gingival crevicular fluid in mouse models with peri-implantitis at different stages.The study included the following three parts:(?)Differential expression of biomarkers in the gingival crevicular fluid between Han and Uyghur people with implants.The gingival crevicular fluid samples from the Han and Uygur subjects with healthy implants and peri-implantitis were subjected to the detections with the Luminex-based Procartaplex multi-cytokine detection kit and the Illumina Miseq sequencing platform.Our results showed that there were differences in the content of some cytokines in the gingival crevicular fluid between the Han and Uyghur subjects,with healthy implants or peri-implantitis.According to the microbial structure,there was no significant difference concerning the healthy implants between the ethnic groups.However,for the subjects with peri-implantitis,significant differences were observed in the dominant flora between the ethnic groups.For the Uyghur subjects with peri-implantitis,the contents of IL-1? were negatively correlated with the dominant flora,including the Acinetobacter,Micrococcus,and Moraxella.(?)Microflora diversity in gingival crevicular fluid of Uyghur subjects with healthy implants and peri-implantitis.Bacterial DNA was extracted from the gingival crevicular fluid of Uyghur subjects with healthy implants and peri-implantitis,and the 16 SrRNA V4-V5 region was sequenced with the Illumina Miseq sequencing platform,to construct the 16 SrRNA gene clone library.The subgingival flora were detected based on the Phylum,Class,Order,Family,and Genus.The OUT dilution curve,species abundance and dominance,alpha diversity,PCoA,and clustering and other comparative analyses were carried out to find out the structural characteristics of subgingival flora of the Uyghur ethnic group.Comparisons based on the Phylum,Class,Order,Family,and Genus levels showed that there were significant differences in the subgingival flora diversity between Uyghur subjects with implants and peri-implantitis.Our results showed that at the level of the door,the class,the eye,the family and the genus,the diversity of the bacterial species of the healthy was found to be different,Moraxella,Micrococcus,and Acinetobacter may represent dominant bacterial species that caused peri-implantitis the this ethnic group.(?)Expression of biomarkers in gingival crevicular fluid from mouse models with peri-implantitis at different stages.Four-week-old C57BL/6NCrSLC mice were selected,and the mouse model of peri-implantitis was established with the silk ligation method.Peri-implantitis was staged according to the staging criteria for human beings with peri-implantitis.Gingival crevicular fluid samples were harvested from these mouse models with peri-implantitis at different stages.The contents of IL-1a,IL-1?,IL-6,IL-17,RANKL,and CRP were detected with the Procartaplex multi-cytokine detection kit(for mice)based on the Luminex platform.The flora structure was analyzed by the sequencing of the samples from peri-implantitis at different stages with the Illumina PE250 sequencing platform.Our results showed that,regardless of the cytokine content or the subgingival flora diversity,there were differences in the indicators for peri-implantitis between different disease stages.In conclusion,our results showed that,there were differences in the expression of some biomarkers between the Han and Uygur patients with peri-implantitis.Differences were observed in the flora structure on the Phylum,Class,Order,Family,and Genus levels,between the Uyghur subjects with healthy implants and peri-implantitis.There were also differences in the expression of biomarkers in the gingival crevicular fluid from mouse models with peri-implantitis at early,middle,and late stages.Compared with the conventional sampling methods(such as blood and local tissue),the gingival crevicular fluid samples have several advantages,i.e.,the samples were easy and economic to collect,transport,and preserve,and the test subjects were more willing to cooperate.Introduction of proteomic,genetic,and microbial biomarkers into the detection and evaluation of peri-implantitis would make the detection results more comprehensive and accurate,providing technical support for the early diagnosis and evaluation of peri-implantitis in clinic,and reduce and/or limit the risk factors.The Luminex platform-based multi-factor detection kit was successfully applied for the simultaneous detection of multiple factors in 25-50 ?L micro-samples,providing true and reliable results within shortened time with reduced effort,which might considered as a novel method for the rapid and accurate measurement of cytokines in the future.The mouse models of peri-implantitis at different stages were established,to obtain the dynamic data concerning the expression of biomarkers in different periods of disease,which provided theoretical support for the prevention,assessment,and treatment of disease at different stages.