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Introduction Previous literature demonstrated the importanceof stacking at least three reversing half-hitches on alternating posts(RHAPs) following arthroscopic knot placement. However, RHAPsconstruction involves looping the suture in either an “overhand” or an“underhand” manner as it relates to the post, which may affect knotsecurity. This study investigated the presently unidentified influenceof different stacking combinations of three RHAPs and suture materialon arthroscopic knot security. Methods Four different RHAPs stacking combinations were tiedwith three different suture materials. Ten knots of each configurationwere tied using each suture material, resulting in 120 evaluated knots.A single load-to-failure test was performed. The mode of failure andmean ultimate clinical failure load were recorded. Results Different overhand/underhand stacking combinations ofthree RHAPs had a statistically significant effect on arthroscopicknot strength and security; however, all combinations surpassed theminimum ultimate clinical failure threshold. Knots constructed witheither Force Fiber® or braided fishing line had mean ultimate clinicalfailure loads of greater than 200 N and most commonly failed due tosuture material breakage (100%, 60 - 80% respectively). Conversely,FiberWire® demonstrated lower mean ultimate clinical failure loadsand had a higher incidence of elongated but intact failure (60 - 90%). Conclusion Different overhand/underhand stacking combinationsof three RHAPs yielded an arthroscopic knot capable of secure tissuefixation. A significant effect was observed for suture materials on theknot strength. This study increases our understanding of suitableRHAPs construction following arthroscopic knot placement that canlead to improving the ultimate clinical failure loads of constructedarthroscopic knots observed between orthopedic surgeons.