Effect Of N-acetylcysteine And Sinomenine On Wear Particle-induced Osteoclastogenesis And Osteolysis

Periprosthetic osteolysis and prosthesis loosening, as a biologic consequence of particulate wear debris limit long-term durability of total joint replacement, although enormous efforts have been made to improve bearing biomaterials, implant designs, and sterilization methods. Recent studies show wear debris such as ultra-high-molecular-weight polyethylene particle, polymethyl- methacrylate particle, kinds of meral particle have a significant role in osteolysis. Wear particles enter the periprosthetic tissue where they are phagocytosed by macrophages,foreign-body giant cell, fibroblast,osteoclast. These cells then release an array of cytokines, such as, IL-1, IL-6, PGE2, TNF-αand other mediators of inflammation that lead to the development of an inflamed granulomatous tissue adjacent to the bone. Eventually, osteoclasts are recruited and/or activated to resorb the bone leading to osteolysis and eventually loosening of the prosthesis. In this article, we evaluate the effect of N-acetylcysteine and sinomenine in the particle-induced osteoclast formation and function, and N-acetylcysteine and sinomenine may be one of effective medicine for clinical treatment of aseptic loosening.PartⅠA study of polymethylmethacrylate particle-induced osteoclastogesisObjective:To study the mechanism PMMA particles can induce osteoclasto- genesis in vitro. Methods: Under the stimulation by polymethylmethacrylate particles, SD rat bone marrow cells were cultured in vitro. The cells were stained with TRAP to observe the morphological variety. The bone resorption pits on bone slices were stained by toluidine blue. Results: PMMA particles induced more TRAP positive osteoclast and bone resorption pits than the control group in vitro. Conclusion : PMMA particles can induce periprosthetic osteolysis, and then lead to aseptic loosening.PartⅡEffect of N-acetylcysteine on wear particle-induced osteoclasto- genesis and osteolysisObjective:To evaluate the effect of N-acetylcysteine on wear particle-induced osteoclastogenesis and osteolysis. Methods: In vitro, cells were cultured 5 days and then exposed to polymethylmethacrylate particles or pretreated with N-acetylcysteine for 1 h prior to stimulation with optimal PMMA particles. Forty-eight hours later, osteoclast counts were determined by tartrate-resistant acid phosphatase staining, and the content of tumor necrosis factor-α, interleukin-1βwas measured with ELISA. Protein expression of the nuclear factor-κB p65 subunit was detected with Western blot. The bone resorption pits on bone slices were stained by toluidine blue. Results: The results showed that N-acetylcysteine inhibited polymethylmethacrylate particle-induced osteoclasto- genic and osteolysis in a dose-dependent manner. The protein levels of TNF-α, IL-1β, and NF-κB was down-regulated by N-acetylcysteine in concentration- dependent manner. Conclusion : N-acetylcysteine may effectively inhibit osteoclastogenesis and suppress wear particle-induced bone resorption. PartⅢEffect of sinomenine on wear particle-induced osteoclastogenesis and osteolysisObjective: To evaluate the effect of Sinomenine on wear particle-induced osteoclastogenesis and osteolysis. Methods: In vitro, cells were cultured 5 days and then exposed to Polymethylmethacrylate particles or pretreated with sinomenine for 1 h prior to stimulation with optimal PMMA particles. Forty-eight hours later, osteoclast counts were determined by tartrate-resistant acid phosphatase staining, and the content of tumor necrosis factor-α, interleukin-1βwas measured with ELISA. Protein expression of the nuclear factor-κB p65 subunit was detected with Western blot. The bone resorption pits on bone slices were stained by toluidine blue. Results: The results showed that sinomenine inhibited PMMA particle-induced osteoclastogenic and osteolysis in a dose-dependent manner. The protein levels of TNF-α, IL-1β, and NF-κB was down-regulated by sinomenine in concentration-dependent manner. Conclusion: Sinomenine may effectively inhibit osteoclastogenesis and suppress wear particle-induced bone resorption.PartⅣEffect of N-acetyl-L-cysteine and sinomenine on wear particle- induced osteoclastogenesis and osteolysisObjective: To evaluate the effect of N-acetyl-L-cysteine and sinomenine on wear particle-induced osteoclastogenesis and osteolysis. Methods: In vitro, cells were cultured 5 days and then exposed to polymethylmethacrylate particles or pretreated with sinomenine and/or N-acetyl-L-cysteine for 1 h prior to stimulation with optimal polymethylmethacrylate particles. Forty-eight hours later, osteoclast counts were determined by tartrate-resistant acid phosphatase staining, and the content of tumor necrosis factor-α, interleukin-1βwas measured with ELISA. Protein expression of the nuclear factor-κB p65 subunit was detected with Western blot. The bone resorption pits on bone slices were stained by toluidine blue. Results: The results showed that sinomenine inhibited polymethylmethacrylate particle-induced osteoclastogenic and osteolysis in a dose-dependent manner, and N-acetylcysteine can increase the inhition of sinomenine. The protein levels of TNF-α, IL-1β, and NF-κB was down- regulated by sinomenine in concentration-dependent manner, and N-acetylcysteine can increase the inhition of sinomenine. Conclusion: N-acetylcysteine has synergetic effect with sinomenine on wear particle-induced osteoclastogenesis and osteolysis.With the upper four experiments, we believe that polymethylmethacrylate particles may induce osteoclastogenesis and osteoclastic osteolysis. N-acetylcysteine and sinomenine may not only inhibt PMMA particle-induced osteoclastogenesis and osteoclastic osteolysis alone, but also N-acetylcysteine has synergetic effect with sinomenine on wear particle-induced osteoclastogenesis and osteolysis.