A Metabonomics Study Of Peri-implant Mucositis Before And After Drug Intervention Based On UHPLC/Q-TOF-MS

Objective: UHPLC/Q-TOF-MS was used to study the metabonomics characteristics of peri-implant sulcus fluid of peri-implant mucositis before and after the drug intervention,and to find out the differential metabolites of peri-implant mucositis before and after treatment.Further search for potential biomarkers for peri-implant mucositis.To provide a new idea and new method for clinical prevention,timely diagnosis and timely treatment of peri-implant mucositis and peri-implantitis.Methods : The peri-implant sulcus fluid were collected(every patient has for dental implant surgery and the crown had be repaired more than half of year),by using the filter paper method.Those were healthy teeth group(group A),healthy implant group(group B),peri-implant mucositis group(group C),and the drug intervention group(group D).There were 15 cases in each group.Randomly selected five samples from each group.The use of UHPLC/Q-TOF-MS for detection and analysis of peri-implant sulcus fluid samples.Using Agilent Masshunter Qualitative B.04.00 Analysis software for the original data preprocessing and normalization.Principal component analysis,partial least square-discriminant analysis and orthogonal partial least square-discriminant analysis were carried out by using SIMCA-P13.0 software.The ions of Variable Importance in the Projection(VIP>1.0)and independent samples t test(P < 0.05)were selected according to the scores plot of the OPLS-DA model.Searching for the differential metabolites between groups.The qualitative method: according to the differential metabolites of accurate molecular weight,retention time,ion mode and two MS information search online database Metlin library and Human metabolome database to make a precise comparison.Results:1.Results of clinical index analysis Four groups of probing depth,peri-implant sulcus fluid,modified plaque index,modified sulcus bleeding index,were statistically significant(P< 0.01).LSD method was used to compare,group A,group B and group D had no statistical difference(P>0.05),the group C was higher than that of group A,group B,group D,and the difference was statistically significant(P<0.01).2.Results of the chromatogram In positive ion mode,the spectrum is mainly in the difference between 6-10min;negative ion mode is mainly reflected in the difference between 9-11 min.3.Results of PCA,PLS-DA and OPLS-DA group A and group B,group B and group C,group C and group D,group B and group D samples in the PCA score plots of the positive and negative ion mode can not be well differentiated,some samples overlap.In the PLS-DA and OPLS-DA score plots,in positive and negative ion modes,group A and group B,group B andgroup C,group C and group D,group B and group D were able to completely separate,show that the metabolic components between the two groups of sample of each comparison group have obvious difference.4.Results of differential metabolitesCompared with healthy teeth group,those differential metabolites in healthy implant group showed a trend of increase,which are:Histidinyl-Aspartate?Dehydrophytosphingosine;those differential metabolites in healthy implant group showed a downward trend,which are:L-Cystine?N-stearoylproline.Compared with healthy implant group,those differential metabolites in peri-implant mucositis group showed a trend of increase,which are:Alanine?3-Indoleacetic Acid ? Xanthine ? Aspartyl-Isoleucine ? Pantothenic Acid ?Prolyl-Valine?1-Linoleoylglycerophosphocholine ?Lysophosphatidylcholine(P-16:0)?Lysophosphatidylethanolamine(18:1(11Z)/P-18:1(9Z));those differential metabolites in peri-implant mucositis group showed a downward trend,which is:1,11-Undecanedicarboxylic acid.Compared with peri-implant mucositis group,those differential metabolites in the drug intervention group showed a downward trend,which are:4-Hydroxydebrisoquine?N,N-Dimethylsphingosine?2-aminohexadecanoic acid ? Urolithin A-3-O-glucuronide ? Lysophosphatidylethanolamine(18:1(11Z)/P-18:1(9Z)).Compared with healthy implant group,those differential metabolites inthe drug intervention group showed a trend of increase,which are: Thiamine pyrophosphate,Lysophosphatidylcholine(20:5(5Z,8Z,11 Z,14Z,17Z)/P-18:1(9Z));those differential metabolites in the drug interven-tion group showed a downward trend,which are:4-Hydroxydebrisoquine?Dihydroceramide C2?Dehydrophytosphingosine ? 2-amino-14,16-dimethyloctadecan-3-ol ?1,11-Undecanedicarboxylic acid?16?-Hydroxynorgestrel.Conclusion:1.Peri-implant mucositis is reversible,the modified plaque index,modified sulcus bleeding index,peri-implant probing depth and peri-implant sulcus fluid amount can be used as common indicators of a check of peri-implant disease.The amount of peri-implant sulcus fluid increased in the inflammatory state.2.The metabonomics technology based on UHPLC/Q-TOF-MS combined with multivariate statistical series is a kind of method for non-invasive,effective,rapid and sensitive differences in metabolic products,to provide new ideas for the pathogenesis of peri-implant mucositis and peri-implant disease.3.The changes of protein metabolism,lipid metabolism,glucose metabolism and citric acid cycle,purine degradation pathway and phospholipid metabolism in the peri-implant mucositis.4.The potential markers of peri-implant mucositis may be Xanthine ?Lysophosphatidylcholine(P-16:0)and Lysophosphatidylethanolamine(18:1(11Z)/P-18:1(9Z)).5.Minocycline hydrochloride and so on in the treatment of peri-implant mucositis after two weeks,although the clinical index of mucosa returned to normal,sugar metabolism,phospholipid metabolism and sphingolipid metabolism are still abnormal.