Effect Of Oral Lichen Planus-associated Myofibroblasts On Migration And Tube-formation Of Vascular Endothelial Cells

BackgroundOral lichen planus(OLP)is a common chronic inflammatory disease of oral mucosa.It's etiology and mechanism is complex.Increasing evidences support that the important role of immune regulation and inflammatory factor mediation on OLP occurrence and development.In OLP inflammatory,there aer abnormal proliferation of blood vessels and dysfunction of vascular endothelial cellsThere are a large number of fibroblasts in the subcutaneous connective tissue of oral mucosa.In addition,stimulated by inflammation,trauma,tumor,and so on,fibroblasts are activated to become myofibroblasts.Myofibroblasts have the ability to promote angiogenesis.Our previous experiments found that 1)the expression of angiogenic related factors increased in the serum and lesion tissues of olp patients;2)?-smooth muscle actin(?-SMA),as a marker protein for myofibroblasts,increasing expression in diseased tissues.Therefore,we hypothesize that the presence of OLP-associated myofibroblast(OLP-MFs)in the OLP matrix,on which we intend to further explore the mechanism of OLP-MFs action on promoting angiogenesis.ObjectiveIntends to explore the methods of isolating and culturing OLP-MFs from the buccal lesion of OLP.To observe the effect of OLP-MFs on the migration and tube-formation of vascular endothelial cells,and to elucidate the role of OLP-MFs in the angiogenesis of oral lichen planus.Method(1)The primary fibroblasts from erosion OLP,non-erosion OLP and normal oral mucosa were obtained by tissue culture.Cultures at passege three were used for the cellular identification of cytokein,vimentin and ?-smooth muscle actin(?-SMA).(2)Interaction models between OLP-MFs and HUVECs were established,compared with the models of NFs and HUVECs,as well as untreated HUVECs.Transwell cell migration assay used to detact the migration of OLP-MFs after co-cultured with HUVECs.Endothelial tube formation assay was used to detect the tube formation of OLP-MFs.Result(1)The expression of cytosolic keratin was negative and vimentin was positive in non-erosive OLP,erosive OLP and normal buccal mucosa tissues.The cytosolic?-SMA expression was negative in normal group,weak positive in non-erosive group OLP and positive in erosive OLP group(2)Considered the migration cell number was 100%in HUVEC group,NFs+HUVEC group was(114.86±8.02)%,OLP-MFs+HUVECs group was(169.83±7.58)%,and OLP-MFs significantly promoted the migration of HUVEC.The tube length of OLP-MFs+HUVEC was(159.90± 12.98)%,and the control groups were(100.38±10.54)%and 100%,respectivelyConclusionThe activated fibroblasts exist in the microenvironment of OLP,and can significantly promote angiogenesis.