| Objective:Analysis the composition of the GCF metabolite around the implant under healthy condition and inflammation. Find out the characteristic metabolic markers. Provide new approaches to the early diagnosis and prevention of clinical peri-implant disease.Methods:We collected 32 clinical cases of dental implantation and receiving upper prosthesis for more than 1 year. Based on the inflammation of tissue around the implant, the cases were divided into 3 groups, which are the healthy implant group(group B), the peri-implant mucositis group(group C)and the peri-implantitis group(group D). The number of cases of each group was 13,10 and 9. Choose healthy natural teeth as control group(group A)in healthy implant group. Check clinical indicators and use Filter paper method to collect the gingival crevicular fluid.Examine the samples with Bruker 600 MHz NMR to get their signal of free induction decay (FID). By importing the signal of FID to the MestReNova-6.1.1 software to carry on the analysis, obtained the nuclear magnetic resonance hydrogen spectrum and sectional integral values. Using principal components analysis (PCA), orthogonal signal correction (OSC) and Partial Least Squares (PLS) to carry out a statistical analysis of the sectional integral values,of which the result is shown in the score chart and the scatter diagram. Analyzing the Variable Importance Plot (VIP) between two different groups and the chemical shift with VIP value larger than 1 contributes more to the distinction of the two groups. Using Non-parametric Tests to analysis the resulting of VIP values which are greater than 1 (when p<0.05, chemical shift difference has statistical meaning. Searching for the metabolic constituents with differences and the potential markers among those groups by retrieving the HMDB database.Results:1. Analytic results of clinical indicatorsmPLI:the index of group C and D are obviously higher than that of group A and B(p<0.05).Group C and D have notable difference(p<0.01). Group A and B have no notable difference(p>0.05).GBI:the index of group C and D are obviously higher than that of group A and B(p<0.05).There is no notable difference between Group C and D (p<0.01) and no notable difference between Group A and B (p>0.05).PPD/PD:the depth of group D is obviously higher than group A,B and C(p<0.01).The probing depth of group D is higher than that of group C(p<0.01).The probing depth of group C is higher than that of group B(p<0.05).The probing depth of group B is higher than that of group A(p<0.01).BL:bone loss of group D is higher than that of group A, B and C(p<0.05). Bone absorption surrounding implant of group B and C has no obvious difference(p>0.05).PISF:group C and D are higher than that of group A and B (p<0.05) There is no obvious difference between group C and D, group A and B (p>0.05).2.1H-NMR spectrum test results:by using nuclear magnetic resonance,44 FID signals are detected. Through fouler transition, phase position and base line regulation,’H-NMR spectrums are very ideal. Spectrum analysis reveals that the differences of spectral peak between the four groups locate mainly in the 3.0-4.2 ppm and 5.0-5.5 ppm.3.Analysis results of PCA and OSC-PLS:By analyzing with PCA and OSC-PLS, we find that there is no notable difference between group A and B, while there is notable difference between group B and C, group B and D, group C and D. In figure of VIP, there are-29 chemical shifts whose VIP value are bigger than 1 between group B and C. There are 21 chemical shifts whose VIP value are bigger than 1 between group B and D. There are 29 chemical shifts whose VIP value are bigger than 1 between group C and D. Making non-parametric test for the chemical shifts whose VIP value are higher than 1, the results as follow:15 chemical shifts value between group B and C have statistical significance(p<0.05).10 chemical shifts value between group B and D have statistical significance (p<0.05).10 chemical shifts value between group C and D have statistical significance(p<0.05).4.Retrieval results in HMDB databaseThe increased metabolites between Group C and group B are leucine, lactic acid, alanine, glutamine, glycine,3-phosphate glycerol, and the declined one is valine. The different metabolites between Group D and group B are isoleucine, alanine, alanine, glutamine,glycine, choline,3-phosphate glycerol and creatine phosphate,which are all increased.The increased metabolites between Group D and group C are very low-density lipoprotein,propanoic acid5alanine,pyruvic acid and guanosine,and the declined ones are thelactic acid,glycoprotein, glycine,3-phosphate glycerol, creatine phosphate.Conclusion:1. Combining the mPLI, GBI, PPD, BL and PISF, can distinguish the inflammation conditions of tissue around implantJPeri-implant sulcular fluid volume is different depend on the tissue around implant healthy or inflammatory, as the degree of inflammation increased, the volume of peri-implant sulcular fluid should be added gradually.Peri-implant sulcular fluid can reflect the conditions of peri-implant tissue.2. The preparation of peri-implant sulcular fluid sample is simple, and the detection method of nuclear magnetic resonance (NMR) is convenient and efficient, the NMR spectra is ideal. Analysis by using PCA and OSC-PLS, We found that the metabolites in peri-implant sulcular fluid between healthy implant group and peri-implant diease group have obvious difference; the gingival crevicular fluid metabolites between healthy natural teeth group and healthy implant group have no obvious difference, indicating that metabonomics method based on 1H-NMR can be used to identify metabolic methodology of peri-implant disease and look for differences in metabolite.3.The different metabolites among peri-implant mucositis, peri-implantitis and healthy implant are mainly involved in three major substance metabolism, illustrating that metabolic status will change when peri-implant tissue status changes.4.The content of glutamine in peri-implant sulcular fluid of peri-implant mucositis and peri-implantitis is higher than that of healthy implant, it may be the marker of inflammation degree of peri-implant tissue.The content of guanosine in peri-implant crevicular fluid of peri-implantitis is higher than peri-implant mucositisis, it may be a potential marker to distinguish them. |
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