A Study Of Screening The Relevant Protein Factors Of Prosthetic Aseptic Loosening In Tha And Expression Of Lentivirus Transduced Antagonist Gene By Stem Cells

[Objective]We systematically reviewed the longitudinal studies of biomarkers in prosthetic aseptic loosening(AL) in arthroplasty to screen the efficient ones and made reasonable choice for biomarker diagnosis study of AL.To compare the metal‐on ‐metal resurfacing arthroplasty(Mo M RA) to the ceramic‐on‐ceramic(Co C) and metal‐on‐polyethylene(Mo P) total hip arthroplasty(THA), a retrospective follow‐up study was done to clarify the relationship between bearing material and osteolysis, make grouping reasonable and collect participants for biomarker diagnosis study of AL.To define the diagnostic value of multiple systemic biomarkers of osteolysis or bone remodeling in early AL.To see if the lentivirus transduced IL‐1 receptor antagonist(Ra) gene was able to be stably expressed by murine bone marrow ‐derived mesenchymal stem cells(BMSCs) in vitro.[Methods]Total 32 longitudinal studies about biomarkers in AL with complete data were included. The study quality was assessed by Oxford Center for Evidence‐based Medicine(CEBM) and the Newcastle‐Ottawa Quality Assessment Scale(NOS) and the evidences were synthesized to show the efficacy and recommendation grade of each biomarker in AL.Total 154 patients(171 hips) were replaced with Mo M RA(group 1 with 27 hips), Co C THA(group 2 with 41 hips) and Mo P THA(group 3 with 103 hips). The index like post‐operative annual osteolysis rate and complication like AL was recorded.Total 105 subjects were recruited including 31 candidates for revision THA(Late AL group), 15 patients who had undergone THA and had clinical and radiographic evidence of AL(Early AL group), 19 patients with no sign of AL(Sable group), and 40 healthy volunteers. Plasma levels of osteoprotegerin(OPG), receptor activator of nuclear factor‐kappa B ligand(RANKL), cross‐linked N‐terminal telopeptide(NTX), procollagen I C‐terminal extension pep‐tide(PICP), tumor necrosis factor‐alpha(TNF‐α), and interleukin(IL)‐1β1 were measured using an immuno ‐enzymatic method. The outcomes of biomarkers were analyzed separately using SPSS software and synthetically using Revman software.The vectors of lentivirus transduced IL‐1Ra/cop GFP gene were constructed and transfected murine bone marrow‐derived mesenchymal stem cell(m BMSCs) to make the Lv.IL‐1Ra.cop GFP/ ‐m BMSCs, which were evalidated by fluorescence microscope, cell viability analysis using CCK‐8 kit and Trypan blue staining, and growth kinetics MTT assay. The expression of IL‐1Ra were showed by RT‐q PCR and Western boltting analysis. ResultsAmong 48 biomarkers only NTX had moderate evidence for AL and the grade of recommendation was B. The IL‐1β, TNF‐a, IL‐8, tartrate resistant acid phosphatase(TRAP), prostaglandin E2 and osteocalcin(OC) had limited evidence and the grade of recommendation was C. After mean 5 years follow‐up, the adjusted annual osteolysis rate was significantly lower in group 1 than in group 2 and 3(both p<0.0001). Mo M RA had least complications without adverse reaction to metal debris(ARMD).Plasma level of IL‐1β and OPG had significant difference between Late AL and non‐AL groups(p<0.05) and only IL‐1β plasma level provided good diagnostic accuracy. There’s significant difference in synthetic analysis of 6 biomarkers not only between Late AL and Stable group, but between Stable and Healthy group(both p=0.02). Heterogeneity of six biomarkers in either comparison was extremely low(both I2 =0).The lentivector transducing IL‐1Ra/cop GFP was well constructed. The m BMSCs showed short proliferation life, but good growth kinetics at early stage and even better viability after IL‐1Ra gene transfection. IL‐1Ra was overexpressed after transfection of m BMSCs. ConclusionsAmong various biomarkers in AL IL‐1β, TNF‐a, IL‐8, TRAP, PGE2, OC and especially NTX had moderate evidence and could be recommended as efficient and reliable markers.All three bearing surfaces of THA, Mo M RA had least osteolysis developing and no ARMD.Plasma IL‐1β had best performance in diagnosis of AL.The multiple biomarkers analysis showed significant change in patients without traditional clinical or radiographic evidence of AL. The Lv.IL‐1Ra.cop GFP/m BMSCs had notable capability of proliferation and were able to effectively express exogenous IL‐1Ra in certain term.