Effects Of The Peri-implantitis On Proliferation And Differentiation Functions Of Osteoblasts

Peri-implantitis is a common complication in oral implantology,which usually occursin the function of the osseointegration of chronic progressive edge of peri-implantitis，can lead to the loss of bone tissue functions. It has been shown in the research thatperi-implant inflammation is a major cause of implant loosening failure..Osteoblasts are the main functional cells of bone formation,in charge of synthesis,secretion and mineralization of bone matrix which plays an important role in theimplant osseointegration.It has been reported in the existing literature,whenperi-implantitis occurs, inflammatory factors affecting the balance between osteoblastand osteoclasts, result the failure of the implant loosening.In this study, jaw bone osteoblasts under the conditions of inflammatorymicroenvironment,it can be observed the ability of proliferation and differentiationinflammatory microenvironment of the biological functions,compared with the normalmandibular osteoblasts. To study the effect of peri-implantitis inflammatorymicroenvironment on the biological function of jaw bone osteoblasts. This study canprovide theoretical basis for guided bone tissueregeneration in inflammatorymicroenvironment.Part I：Objective:Collect bone tissue form clinical surgery, to cultureinflammatorytissue-derived osteoblasts and normal tissue-derived osteoblasts.Methods: Collect PISF(Peri-implant sulcus fluid)form implant failed withperi-implantitis patient. Collect GCF (gingival crevicular fluid) from third molarsurgery patient. ALP and AST levels in PISF and GCF were determined by biochemicalanalyzer. TNF-α and IL-6levels in PISF and GCF were determined by ELISA. Results: ALP,AST,TNF-α and IL-6levels in PISF was higher than GCF.Conclusion:ALP,AST,TNF-α and IL-6levels can be an important evaluate indexes withperi-implant infection.Part II:Objective:To isolate, culture and purify inflammatory tissue-derived osteoblasts andnormal tissue-derived osteoblasts, identify the proliferation of two different tissuesources osteoblasts.Identify two different tissue-derived osteoblasts through ALPstaining and alizarin red staining.Methods: Collect inflammatory bone tissue form implant failed with peri-implantitispatient. Collect healthy bone tissue from third molar surgery patient.To adopt theimproved method of tissue blocks,isolate and culture the inflammatory tissue-derivedosteoblasts and normal tissue-derived osteoblasts, which were purified by differentialadhesion method.The purified osteoblasts were identified through ALP staining andalizarin red staining.The proliferative activity of two different tissue-derived osteoblastswas evaluated through MTT assay.Results: ALP staining and alizarin red staining showed positive on the basis ofpurified inflammatory tissue-derived osteoblasts and normal tissue-derived osteoblasts.In the proliferative activity of the cells, the two different tissue-derived cells on thefourth day began to show statistically significant differences,osteoblasts fromperi-implantitis became lower than that of normal tissue,which has been shown in theMTT assay.Conclusion: The improved method of tissue culture and differential adhesion methodcan obtain the purified inflammatory tissue-derived osteoblasts and normaltissue-derived osteoblasts. Peri-implantitis inflammatory microenvironment candecreasethe proliferation activity of osteoblasts.Part III:Objective: To identify the differentiation capacity of two different osteoblasts tissuesources. Methods: Osteoblasts tissue were collected from two different sources. Expression ofOCN, Runx2and Col-1were examined by RT-PCR technique. OCN protein levels weredetermined by Western blots.The results: Expression of OCN, Runx2and Col-1significantly decreased ininflammatory tissue compared with the normal. OCN protein levels was also decreasedin inflammatory tissue.Conclusion: Differentiation of osteoblasts was reduced in peri-implantitisinflammatory microenvironment.