不同磨损颗粒对体外培养人外周血单核细胞的影响
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摘要
背景:人工关节置换后正常的行走活动会产生大量的钛铝钒合金(Titanium6Aluminium4Vanadium,Ti6Al4V)、聚甲基丙烯酸甲酯(Polymethylmethacrylate,PMMA)、超高分子量聚乙烯(Ultra-High-MolecularWeight-Polyethylene,UHMWPE)、钴铬合金(Chromium-Cobalt Alloy,Co-Cr)等磨损颗粒,激活单核细胞释放大量的炎性细胞因子和趋化因子,诱发骨溶解和形成假性肉芽肿等一系列生物学级联反应,最终导致假体发生无菌性松动。目的:探讨Ti6Al4V、PMMA、UHMWPE和Co-Cr颗粒对体外培养人外周血单核细胞分泌肿瘤坏死因子α、白细胞介素1和白细胞介素6的影响及诱导骨溶解引起无菌性松动的作用机制。方法:将人外周血单核细胞分别与不同浓度的Ti6Al4V、PMMA、UHMWPE和Co-Cr颗粒进行共培养,72 h后提取细胞上清液,用ELISA法检测肿瘤坏死因子α、白细胞介素1和白细胞介素6的表达水平,观察各种颗粒浓度和理化性质对外周血单核细胞活性的影响。结果与结论:(1)各组人外周血单核细胞存活率均大于95%,仅有Co-Cr组颗粒/细胞比值为1 000时单核细胞存活率降至87%;(2)当颗粒/细胞比值为10,100,1000时,活化的单核细胞分泌肿瘤坏死因子α、白细胞介素1和白细胞介素6的水平与原始幼稚单核细胞(颗粒/细胞比值为0时)相比显著增高,差异有显著性意义(P <0.05);(3)Ti6Al4V组、PMMA组和UHMWPE组中随颗粒浓度增加,单核细胞分泌肿瘤坏死因子α、白细胞介素1和白细胞介素6出现不同程度的增强。Co-Cr组中当颗粒/细胞比值达到1 000时却出现肿瘤坏死因子α、白细胞介素1和白细胞介素6的表达水平降低;(4)在骨溶解过程中Ti6Al4V颗粒的功能生物学活性最强,其次是Co-Cr和PMMA颗粒,UHMWPE颗粒最低弱;Ti6Al4V、PMMA、UHMWPE和Co-Cr颗粒在骨溶解过程中对外周血单核细胞的影响存在剂量依赖性。
BACKGROUND: After artificial joint replacement, daily activities lead to the production of a large number of titanium 6 aluminium 4 vanadium(Ti6 Al4 V), polymethylmethacrylate(PMMA), ultra-high-molecular-weight-polyethylene(UHMWPE) and chromium-cobalt alloy(Co-Cr) particles, activating monocytes to release a large number of inflammatory cytokines and chemokines, including a series of biological cascade reactions, such as inducing bone dissolution and forming pseudo-granuloma, and eventually leading to aseptic loosening of the prosthesis. OBJECTIVE: To investigate the effect of Ti6 Al4 V, PMMA, UHMWPE and Co-Cr particles on tumor necrosis factor-α, interleukin-1 and interleukin-6 in the human peripheral blood monocytes cultured in vitro and to explore the underlying mechanism of aseptic loosening of the prosthesis by inducing bone dissolution. METHODS: Human peripheral blood mononuclear cells were cultured with Ti6 Al4 V, PMMA, UHMWPE and Co-Cr particles at different concentrations. After 72 hours of culture, ELISA technique was utilized to quantify the secretion of different cytokines includes tumor necrosis factor-α, interleukin-1 and interleukin-6. We observed the influence of different wear particle concentrations and particle properties on the viability of human peripheral blood mononuclear cells. RESULTS AND CONCLUSION: Cell viability exceeded 95% for all groups, except for 87% in the Co-Cr group when the particle/cell ratio was 1 000. Compared with the naive mononuclear cells(particle/cell ratio of 0), the expression of tumor necrosis factor-α, interleukin-1 and interleukin-6 in the activated mononuclear cells increased significantly when the particle/cell ratio was 10, 100 and 1 000(P < 0.05). Ti6 Al4 V, PMMA and UHMWPE particles stimulated human mononuclear cells to overexpress tumor necrosis factor-α, interleukin-1 and interleukin-6, which were positively correlated with the concentration of particles. However, when the Co-Cr particle/cells ratio was 1 000, the expression levels of tumor necrosis factor-α, interleukin-1 and interleukin-6 were decreased. During the osteolysis reaction, Ti6 Al4 V particles had strongest biological activity, followed by Co-Cr and PMMA particles, and the biological activity of UHMWPE particles was lowest. Ti6 Al4 V, PMMA, UHMWPE and Co-Cr particles showed dose-dependent effects on the human peripheral blood mononuclear factors during the osteolysis.
BACKGROUND: After artificial joint replacement, daily activities lead to the production of a large number of titanium 6 aluminium 4 vanadium(Ti6 Al4 V), polymethylmethacrylate(PMMA), ultra-high-molecular-weight-polyethylene(UHMWPE) and chromium-cobalt alloy(Co-Cr) particles, activating monocytes to release a large number of inflammatory cytokines and chemokines, including a series of biological cascade reactions, such as inducing bone dissolution and forming pseudo-granuloma, and eventually leading to aseptic loosening of the prosthesis. OBJECTIVE: To investigate the effect of Ti6 Al4 V, PMMA, UHMWPE and Co-Cr particles on tumor necrosis factor-α, interleukin-1 and interleukin-6 in the human peripheral blood monocytes cultured in vitro and to explore the underlying mechanism of aseptic loosening of the prosthesis by inducing bone dissolution. METHODS: Human peripheral blood mononuclear cells were cultured with Ti6 Al4 V, PMMA, UHMWPE and Co-Cr particles at different concentrations. After 72 hours of culture, ELISA technique was utilized to quantify the secretion of different cytokines includes tumor necrosis factor-α, interleukin-1 and interleukin-6. We observed the influence of different wear particle concentrations and particle properties on the viability of human peripheral blood mononuclear cells. RESULTS AND CONCLUSION: Cell viability exceeded 95% for all groups, except for 87% in the Co-Cr group when the particle/cell ratio was 1 000. Compared with the naive mononuclear cells(particle/cell ratio of 0), the expression of tumor necrosis factor-α, interleukin-1 and interleukin-6 in the activated mononuclear cells increased significantly when the particle/cell ratio was 10, 100 and 1 000(P < 0.05). Ti6 Al4 V, PMMA and UHMWPE particles stimulated human mononuclear cells to overexpress tumor necrosis factor-α, interleukin-1 and interleukin-6, which were positively correlated with the concentration of particles. However, when the Co-Cr particle/cells ratio was 1 000, the expression levels of tumor necrosis factor-α, interleukin-1 and interleukin-6 were decreased. During the osteolysis reaction, Ti6 Al4 V particles had strongest biological activity, followed by Co-Cr and PMMA particles, and the biological activity of UHMWPE particles was lowest. Ti6 Al4 V, PMMA, UHMWPE and Co-Cr particles showed dose-dependent effects on the human peripheral blood mononuclear factors during the osteolysis.
引文
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[17]Atkins GJ,Haynes DR,Howie DW,et al.Role of polyethylene particles in peri-prosthetic osteolysis:A review.World J Orthop.2011;2(10):93-101.
[18]Matthews JB,Besong AA,Green TR,et al.Evaluation of the response of primary human peripheral blood mononuclear phagocytes to challenge with in vitro generated clinically relevant UHMWPE particles of known size and dose.J Biomed Mater Res.2000;52(2):296-307.
[19]Shanbhag AS,Jacobs JJ,Black J,et al.Human monocyte response to particulate biomaterials generated in vivo and in vitro.JOrthop Res.1995;13(5):792-801.
[20]Shanbhag AS,Jacobs JJ,Black J,et al.Effects of particles on fibroblast proliferation and bone resorption in vitro.Clin Orthop Relat Res.1997;(342):205-217.
[21]Haynes DR,Rogers SD,Hay S,et al.The differences in toxicity and release of bone-resorbing mediators induced by titanium and cobaltchromium-aloy wear particles.J Bone Joint Surg Am.1993;75(6):825-834.
[22]Sethi RK,Neavyn MJ,Rubash HE,et al.Macrophage response to cross-linked and conventional UHMWPE.Biomaterials.2003;24(15):2561-2573.
[23]Yamanaka Y,Karuppaiah K,Abu-Amer Y.Polyubiquitination events mediate polymethylmethacrylate(PMMA)particle activation of NF-kappaB pathway.J Biol Chem.2011;286(27):23735-23741.
[24]González O,Smith RL,Goodman SB.Effect of size,concentration,surface area,and volume of polymethylmethacrylate particles on human macrophages in vitro.J Biomed Mater Res.1996;30(4):463-473.
[25]Sabbatini M,Piffanelli V,Boccafoschi F,et al.Different apoptosis modalities in periprosthetic membranes.J Biomed Mater Res A.2010;92(1):175-184.
[26]Catelas I,Petit A,Zukor DJ,et al.TNF-alpha secretion and macrophage mortality induced by cobalt and chromium ions in vitro-qualitative analysis of apoptosis.Biomaterials.2003;24(3):383-391.
[27]Schalock PC,MennéT,Johansen JD,et al.Hypersensitivity reactions to metallic implants-diagnostic algorithm and suggested patch test series for clinical use.Contact Dermatitis.2012;66(1):4-19.
[28]Tsaousi A,Jones E,Case CP.The in vitro genotoxicity of orthopaedic ceramic(Al2O3)and metal(CoCr alloy)particles.Mutat Res.2010;697(1-2):1-9.
[29]VanOs R,Lildhar LL,Lehoux EA,et al.In vitro macrophage response to nanometer-size chromium oxide particles.J Biomed Mater Res B Appl Biomater.2014;102(1):149-159.
[2]Del Buono A,Denaro V,Maffulli N.Genetic susceptibility to aseptic loosening following total hip arthroplasty:a systematic review.Br Med Bull.2012;101:39-55.
[3]Gudena R,Kuna S,Pradhan N.Aseptic loosening of total hip replacement presenting as an anterior thigh mass.Musculoskelet Surg.2013;97(3):247-249.
[4]Landgraeber S,Wegner A,Canbay A,et al.Serum levels of adiponectin in patients with aseptic loosening after total hip replacement.J Biomed Mater Res A.2010;93(2):748-752.
[5]Ollivere B,Darrah C,Brankin RC,et al.The continued value of clinical and radiological surveillance:the Charnley Elite Plus hip replacement system at 12 years.J Bone Joint Surg Br.2009;91(6):720-724.
[6]Bozic KJ,Kurtz SM,Lau E,et al.The epidemiology of revision total hip arthroplasty in the United States.J Bone Joint Surg Am.2009;91(1):128-133.
[7]Jiang Y,Jia T,Wooley PH,et al.Current research in the pathogenesis of aseptic implant loosening associated with particulate wear debris.Acta Orthop Belg.2013;79(1):1-9.
[8]Jiang Y,Jia T,Gong W,et al.Effects of Ti,PMMA,UHMWPE,and Co-Cr wear particles on differentiation and functions of bone marrow stromal cells.J Biomed Mater Res A.2013;101(10):2817-2825.
[9]Ingham E,Fisher J.Biological reactions to wear debris in total joint replacement.Proc Inst Mech Eng H.2000;214(1):21-37.
[10]Zhang K,Yang SY,Yang S,et al.Different influence of Ti,PMMA,UHMWPE,and Co-Cr particles on peripheral blood monocytes during periprosthetic inflammation.J Biomed Mater Res A.2015103(1):358-364.
[11]Lin TH,Tamaki Y,Pajarinen J,et al.Chronic inflammation in biomaterial-induced periprosthetic osteolysis:NF-κB as a therapeutic target.Acta Biomater.2014;10(1):1-10.
[12]Catelas I,Jacobs JJ.Biologic activity of wear particles.Instr Course Lect.2010;59:3-16.
[13]Lübbeke A,Garavaglia G,Barea C,et al.Influence of patient activity on femoral osteolysis at five and ten years following hybrid total hip replacement.J Bone Joint Surg Br.2011;93(4):456-463.
[14]Chiba J,Rubash HE,Kim KJ,et al.The characterization of cytokines in the interface tissue obtained from failed cementless total hip arthroplasty with and without femoral osteolysis.Clin Orthop Relat Res.1994;(300):304-312.
[15]Shanbhag AS,Kaufman AM,Agarwal S,et al.Critical insights into osteolysis using protein microarrays:The importance of IL-6 and T-cell Activation.Trans Orthop Res Soc.2005;30:148.
[16]Kaufman AM,Alabre CI,Rubash HE,et al.Human macrophage response to UHMWPE,TiAlV,CoCr,and alumina particles:analysis of multiple cytokines using protein arrays.J Biomed Mater Res A.2008;84(2):464-474.
[17]Atkins GJ,Haynes DR,Howie DW,et al.Role of polyethylene particles in peri-prosthetic osteolysis:A review.World J Orthop.2011;2(10):93-101.
[18]Matthews JB,Besong AA,Green TR,et al.Evaluation of the response of primary human peripheral blood mononuclear phagocytes to challenge with in vitro generated clinically relevant UHMWPE particles of known size and dose.J Biomed Mater Res.2000;52(2):296-307.
[19]Shanbhag AS,Jacobs JJ,Black J,et al.Human monocyte response to particulate biomaterials generated in vivo and in vitro.JOrthop Res.1995;13(5):792-801.
[20]Shanbhag AS,Jacobs JJ,Black J,et al.Effects of particles on fibroblast proliferation and bone resorption in vitro.Clin Orthop Relat Res.1997;(342):205-217.
[21]Haynes DR,Rogers SD,Hay S,et al.The differences in toxicity and release of bone-resorbing mediators induced by titanium and cobaltchromium-aloy wear particles.J Bone Joint Surg Am.1993;75(6):825-834.
[22]Sethi RK,Neavyn MJ,Rubash HE,et al.Macrophage response to cross-linked and conventional UHMWPE.Biomaterials.2003;24(15):2561-2573.
[23]Yamanaka Y,Karuppaiah K,Abu-Amer Y.Polyubiquitination events mediate polymethylmethacrylate(PMMA)particle activation of NF-kappaB pathway.J Biol Chem.2011;286(27):23735-23741.
[24]González O,Smith RL,Goodman SB.Effect of size,concentration,surface area,and volume of polymethylmethacrylate particles on human macrophages in vitro.J Biomed Mater Res.1996;30(4):463-473.
[25]Sabbatini M,Piffanelli V,Boccafoschi F,et al.Different apoptosis modalities in periprosthetic membranes.J Biomed Mater Res A.2010;92(1):175-184.
[26]Catelas I,Petit A,Zukor DJ,et al.TNF-alpha secretion and macrophage mortality induced by cobalt and chromium ions in vitro-qualitative analysis of apoptosis.Biomaterials.2003;24(3):383-391.
[27]Schalock PC,MennéT,Johansen JD,et al.Hypersensitivity reactions to metallic implants-diagnostic algorithm and suggested patch test series for clinical use.Contact Dermatitis.2012;66(1):4-19.
[28]Tsaousi A,Jones E,Case CP.The in vitro genotoxicity of orthopaedic ceramic(Al2O3)and metal(CoCr alloy)particles.Mutat Res.2010;697(1-2):1-9.
[29]VanOs R,Lildhar LL,Lehoux EA,et al.In vitro macrophage response to nanometer-size chromium oxide particles.J Biomed Mater Res B Appl Biomater.2014;102(1):149-159.