Effects of cobalt and chromium ions on glycolytic flux and the stabilization of hypoxia‐inducible factor‐1α in macrophages in vitro

Implant wear and corrosion have been associated with adverse tissue reactions that can lead to implant failure. Wear and corrosion products are therefore of great clinical concern. For example, Co²⁺ and Cr³⁺ originating from CoCrMo‐based implants have been shown to induce a proinflammatory response in macrophages in vitro. Previous studies have also shown that the polarization of macrophages by some proinflammatory stimuli is associated with a hypoxia‐inducible factor‐1α (HIF‐1α)‐dependent metabolic shift from oxidative phosphorylation (OXPHOS) towards glycolysis. However, the potential of Co²⁺ and Cr³⁺ to induce this metabolic shift, which plays a determining role in the proinflammatory response of macrophages, remains largely unexplored. We recently demonstrated that Co²⁺, but not Cr³⁺, increased oxidative stress and decreased OXPHOS in RAW 264.7 murine macrophages. In the present study, we analyzed the effects of Co²⁺ and Cr³⁺ on glycolytic flux and HIF‐1α stabilization in the same experimental model. Cells were exposed to 6 to 24 ppm Co²⁺ or 50 to 250 ppm Cr³⁺. Glycolytic flux was determined by analyzing extracellular flux and lactate production, while HIF‐1α stabilization was analyzed by immunoblotting. Results showed that Co²⁺, and to a lesser extent Cr³⁺, increased glycolytic flux; however, only Co²⁺ acted through HIF‐1α stabilization. Overall, these results, together with our previous results showing that Co²⁺ increases oxidative stress and decreases OXPHOS, suggest that Co²⁺ (but not Cr³⁺) can induce a HIF‐1α‐dependent metabolic shift from OXPHOS towards glycolysis in macrophages. This metabolic shift may play an early and pivotal role in the inflammatory response induced by Co²⁺ in the periprosthetic environment.     

In Vivo Corrosion of Sleeved Ceramic Femoral Heads: A Retrieval Study

BackgroundThe purpose of this study was to evaluate a series of retrieved sleeved ceramic femoral heads used in total hip arthroplasty (THA) and determine qualitative and quantitative damage and corrosion patterns.MethodsAn IRB-approved implant retrieval database was utilized to identify all sleeved ceramic femoral heads collected from 1995 to 2004. There were 16 implants with an average duration of in situ of 70 months (range, 13-241 months). The femoral stem was known in 14 cases and was titanium alloy in each of those cases. None were revised for metal-related complications. Ten implants (63%) were from primary THAs, and 6 (38%) were from revision THAs. Damage and corrosion were qualitatively graded using a modified Goldberg method. A quantitative assessment was performed with a coordinate measurement machine (CMM).ResultsAmong the 16 retrieved implants, 1 (6%) demonstrated severe Grade 4 corrosion, 5 (31%) had moderate Grade 3 corrosion, 5 (31%) had mild Grade 2 corrosion, and 5 (31%) had no visible corrosion at the inner sleeve that interfaces with the stem trunnion. The only case of grade 4 corrosion occurred in the only head-sleeve in the study that was not factory assembled and was mated with a titanium molybdenum zirconium ferrous (TMZF) alloy stem. The mean maximum linear corrosion depth at the taper interface, as measured by the CMM, was 7.7 microns (range, 0.9-32.9 microns).ConclusionThis study is the first to quantify corrosion at the titanium interface of sleeved ceramic femoral heads. Potentially clinically significant damage and corrosion patterns were observed in a few failed retrievals; however, the majority of cases demonstrated minimal or no damage.     

Pseudotumors in Small-Head Metal-on-Metal Total Hip Arthroplasties at a Minimum Follow-Up of 20 Years

BackgroundMetal-on-metal (MOM) surfaces in total hip arthroplasty (THA) have been used widely. Serum cobalt and chromium levels have been the standard investigation for follow-up examinations, but magnetic resonance imaging (MRI) with metal artifact reducing sequences has shown good results in detecting pseudotumors. The aim of this study is to survey a significant correlation among MRI findings, serum metal levels, and clinical scores in patients with small-head MOM implants and if serum cobalt and chromium levels are sufficient in detecting patients with pseudotumors in the long-term follow-up.MethodsAt a minimum follow-up of 20 years, 26 patients (29 THAs) of the original 98 patients (105 THAs) included in this study between November 1992 and May 1994 were available for follow-up examination. Clinical scores, serum metal ion levels, and MRIs were obtained.ResultsWe found mean serum cobalt levels of 1.87 μg/L (±3.44) and chromium levels of 2.23 μg/L (±2.96) and very good clinical and functional results (mean Harris Hip Score 88.6) in the long-term follow-up. Pseudotumors were detected in MRIs of 21 hips. There were no significant differences between patients with or without pseudotumors regarding serum metal levels and the correlation for clinical outcome scores, demographic data, and cup inclination. The cumulative rate of survival was still at 91.4% at 22.8 years.ConclusionThis study presents the first published data on small-head MOM hips, comparing metal ion levels, pseudotumors, clinical, and radiological results in a follow-up period of more than 20 years and reveals that serum metal levels are not significantly higher in patients with pseudotumors.Level of EvidenceTherapeutic Level III     

Survivorship of Ceramic-on-Ceramic Total Hip Arthroplasty With Metal-Backed Acetabular Liners at 10 Years

BackgroundCeramic-on-ceramic bearings are becoming increasingly popular in primary total hip arthroplasty (THA). To enhance ceramic-on-ceramic liner exchange in case of revision surgery, metal-backed liner systems have been proposed. Little is known about the clinical performance of these implants. The purpose of this study is to evaluate a metal-backed liner implant system for primary THA.MethodsA total of 422 patients (with 468 consecutive THAs) were followed over a mean period of 10 years. All arthroplasties were performed with a cementless stem, a press-fit cup, and a metal-backed liner system. Surgical and clinical data, complications, and revisions were analyzed. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) before surgery and at 1 and 10 years after surgery were compared.ResultsThe overall 10-year implant survival rate was 93.8%. The survival rate was 97.0% for heads and liners, 97.5% for stem, and 99.3% for acetabular cup. The most common reason for revision was ceramic breakage (2.4%) of the third-generation (BIOLOX forte) acetabular liner. Mean WOMAC score improved significantly from 50.1 before surgery to 13.2 at 1 year after surgery. There was no difference in WOMAC scores between surgical approach and type of bearing at 1 and 10 years after surgery.ConclusionTHA using cementless stem, press-fit cup, and metal-backed liner system provides satisfactory long-term outcomes, with revision rate comparable to that with other systems available in the market. The metal-backed liner system has low risk of mal-seating. Third-generation ceramic liners should be avoided as they seem to be more prone to breakage.     

双相生物钙磷陶瓷生物相容性及异位骨诱导的实验研究

目的检测运用羟基磷灰石及β-磷酸三钙制备的双相钙磷陶瓷的生物相容性及其异位骨诱导效率。方法采用化学共沉淀法及过氧化氢发泡法,将羟基磷灰石及β-磷酸三钙以6∶4的比例在1 100 ℃条件下烧结3 h获得双相钙磷陶瓷,利用X线衍射评估材料组成成分。分离SD大鼠骨髓间充质干细胞,接种于双相钙磷陶瓷,通过扫描电镜、鬼笔环肽及DAPI染色观察细胞的黏附,CCK8法评估细胞增殖,碱性磷酸酶测定法评估骨髓间充质干细胞的碱性磷酸酶表达活性。将不含骨髓间充质干细胞的双相钙磷陶瓷置入比格犬竖脊肌内,于4、8、12周对样本行大体检测、组织染色,测算新骨生成率,从而评估双相钙磷陶瓷的异位骨诱导效率。结果成功制备双相钙磷陶瓷,X线衍射分析可见羟基磷灰石及β-磷酸三钙特异性的衍射峰。扫描电镜可见双相钙磷陶瓷表面广泛分布大孔及连通孔,孔壁粗糙不平,孔内可见均匀分布的微孔。鬼笔环肽及DAPI染色显示,骨髓充质干细胞在材料表面伸展黏附,共培养后逐渐从不规则形转变为均一的长梭形。CCK8法提示共培养后第1天,细胞活力降低,而第3、4、5、7天,细胞增殖活力逐渐增加。碱性磷酸酶活性检测提示,与对照组相比,共培养后第1、7天,双相钙磷陶瓷组的骨髓间充质干细胞可分泌更多的碱性磷酸酶。双相钙磷陶瓷顺利置入比格犬竖脊肌内,术后8周材料孔隙内可见骨样组织沉积,术后12周大孔成骨比例为0.77±0.11,孔内成骨面积比例为0.71±0.14。结论双相钙磷陶瓷具有良好的生物相容性及异位骨诱导效率。     

微量元素Zn,Cu,Fe与高血压关系的病例对照研究

目的探讨原发性高血压的发病因素。方法病例对照研究方法,分析了微量元素Ｚｎ、Ｃｕ、Ｆｅ与高血压的关系。结果Ｃｕ、Ｆｅ、Ｚｎ／Ｃｕ、Ｚｎ／Ｆｅ、Ｆｅ／Ｃｕ与高血压有一定关系,其中以Ｃｕ、Ｆｅ／Ｃｕ与高血压的关系更为密切。同时单相关分析说明Ｆｅ对收缩压和舒张压有正性作用,Ｚｎ／Ｆｅ对收缩压有负性联系。结论Ｚｎ／Ｃｕ、Ｚｎ／Ｆｅ、Ｆｅ／Ｃｕ对血压的作用较独立的Ｚｎ、Ｃｕ、Ｆｅ显著     

Peptide formation in the presence of metal ion protecting groups

A quantitative study of the degree of racemization induced by the [(NH₃)₅Co-(III)-] protecting group when bound to the C-terminal of the amino acids Leu, Phe, and His, as has been carried out. Racemization was determined by forming the diastereomeric cobalt dipeptides [(Leu)(AA)Co(III)(NH3)₅] where AA = L-Leu, L-Phe, and L-His; after cobalt removal (using NaBH₄), the peptide diastereomers were analyzed quantitatively using an amino acid analyzer. No racemization was observed within experimental error (0.3%) as a result of the substitution of the [(NH3)₅Co(III)-] group on the amino acids and peptides studied.     

环丙沙星荧光特性的研究

目的:建立荧光测定环丙沙星含量的方法,对几种金属离子(Fe~(3+),Cu~(2+),Zn~(2+))与环丙沙星的络合反应及其对荧光特性的影响作初步的研究。方法:用日立荧光分光光度计测定,测量波长λ_(ex)=277 nm,λ_(em)=450nm。测定时的最佳pH为2.2。结果:环丙沙星浓度在0.1～0.8mg·L~(-1)范围内与荧光强度呈线性关系,相关系数为0.9999,检测限为4.5×10~(-3) mg·L~(-1)。结论:该法方便、快速,可用于直接测定环丙沙星含量,或用于微量分析。     

Biophysical studies and anti‐growth activities of a peptide,a certain analog and a fragment peptide derived from α‐fetoprotein

Abstract: A chemically synthesized 34‐amino acid peptide, an analog, and a fragment of the peptide have been purified and studied. Biophysical studies were carried out to determine some of the metal ion binding properties of the original peptide and an analog of this parent peptide, in which the two histidine residues were replaced by alanines. As shown by visible absorption spectroscopy, Co (II) forms a complex with the parent peptide, but not with the analog peptide, and one or two histidines in the parent peptide are ligands for Co (II) ion binding. The effects on disulfide bond formation in the peptide by Zn (II) and Co (II) ions were also examined for this analog. Anti‐growth assays were performed using the original cysteine‐containing peptide with Zn (II) ion complexed to the peptide through the two cysteine residues. These rat uterine growth assays showed that the complexing of Zn (II) ion to the peptide maintained the anti‐growth activity of the peptide, while gel‐filtration experiments showed the zinc ions maintained the peptide in its anti‐growth form indefinitely in solution. A saliently important part of this research was the discovery that a fragment of the peptide consisting of a middle sequence of 14 amino acids was found to have significant anti‐growth activity in the rat uterine assay. Its activity suggested that this fragment might be considered a viable candidate for testing in anti‐cancer protocols.     

Trace Metals' abnormalities in hemodialysis patients: relationship with medications

A multicenter collaborative study was performed to investigate the prevalence of abnormal blood contents of 6 trace metals, copper (Cu), zinc (Zn), aluminum (Al), lead (Pb), cadmium (Cd), and mercury (Hg), in hemodialysis (HD) patients and to analyze their relationship with the medications, such as CaCO3, Ca acetate, Al containing phosphate-binding agents, 1,25-dihydroxy vitD3, 1-hydroxy vitD3, and erythropoietin (EPO), as well as hematocrit level, by chi-square statistics. From 6 medical centers in Taiwan, we included 456 patients in maintenance HD for more than 4 months for this study, and they had continued the previously mentioned medications for at least 3 months. Blood samples were collected before initiating HD, and atomic absorption spectrophotometry was used to measure plasma levels of Cu, Zn, and Al as well as whole blood levels of Pb, Cd, and Hg. Three hundred seventy-five (78%) of the HD patients had low plasma Zn levels, that is, <800 microg/L, and the mean (+/-SD) concentration was 705.8 (+/-128.23) microg/L in all subjects. One hundred forty-one (31%) of the HD patients had high plasma Al, that is, >50 microg/L, and the mean (+/-SD) was 44.30 (+/-28.28) microg/L in all subjects. Three hundred thirty-three (73%) of the dialysis patients had high Cd levels, that is, >2.5 microg/L, and the mean (+/-SD) was 3.32 (+/-1.49) microg/L in all subjects. The majority of HD patients had normal blood levels of Cu, PB, and Hg. Only 21 (4. 6%), 5 (1.1%), and 3 (0.06%) patients had elevated blood levels of Cu, Pb, and Hg, respectively. Their mean (+/-SD) blood concentration of Cu, Pb, and Hg were 1,049.78 (+/-233.25) microg/L, 7.45 (+/-3.95) microg/dL, and 3.17 (+/-25.56) microg/L, respectively. Three patients had elevated plasma Hg concentrations, that is, 546, 12.6, and 24.0 microg/L, respectively. In the 152 normal healthy age and sex matched control group, the blood levels of Al, Cd, and Pb were all significantly lower than the HD patients. However, the levels of Cu and Zn were higher in the control group. The Hg level was not significantly different in both groups. There was no statistical difference between patients with normal and abnormal blood levels of trace metals in various medications except Al containing phosphate binder. The Al containing phosphate binder users had significantly higher plasma Al levels (54.71 +/- 26.70 versus 41.15 +/- 28.03 microg/L, p < 0.001) and hematocrit levels (29.61 +/- 4.61 versus 27. 81 +/- 3.91, p < 0.0005). There was no statistical correlation between erythropoietin (EPO) dose and hematocrit level in these patients. In conclusion, the blood level of trace metals of these HD patients except Al was not related to their medications. However, caution must be exercised in interpreting this result as dose and duration of medication; efficiency of HD and water treatment may play an important role. Otherwise, environmental factors, diet, and the aging process may contribute to the trace metal burden in uremia. Thus, Zn and Cu are abundant in seafood, and Cd is abundant in contaminated plants such as rice.