Original Article Spatula DD. 2011; 1(2):81-89 EDTA chelation therapy in the treatment of toxic metals exposure Toksik metallere maruz kalım tedavisinde EDTA şelasyon terapisi Nina Mikirova1, Joseph Casciari1, Ronald Hunninghake1, Neil Riordan1,2 1Riordan Clinic, Wichita, Kansas, USA. 2Medistem Inc, San Diego, California, USA SUMMARY BACKGROUND: Metal induced toxicity with wide range of physiological, biochemical and behavioral dysfunctions was reported in many studies. The chelation has been used for treatment to toxic metals’ exposure for many years. In our current clinical study, we compared different chelation protocols and two forms of EDTA (sodium calcium edetate and sodium edetate) in treatments of toxic metal exposure. METHODS: A 24 h urine samples were collected from each subject before and after treatment by Ca-EDTA or Na-EDTA. The levels of toxic and essential metals were measured by Atomic Absorption Spectrometer (AAS) with graphite furnace and by Inductively Coupled Plasma Atomic Emission Spectrometer (ICP-AES). The cellular levels of ATP were determined by ATP-bioluminescence assay. Mitochondrial potential was measured by fluorometer and flow-cytometer after cells’ staining by mitochondrial dye. RESULTS: Data from over 600 patients with a variety of complaints, but not acute toxic mineral exposure, given chelation therapy with sodium EDTA or calcium EDTA, were analyzed. Ca-EDTA and Na-EDTA at intravenous infusions of 3 g per treatment were equally effective in removing lead from the body, while Ca-EDTA was more effective in aluminum removal. The removal of lead was dose dependent but non-linear. Chelation by different doses of Na-EDTA (3 g and 1 g) resulted in mean difference in lead urine less than 50%. In addition, deficiencies in essential minerals correlated with greater pre-treatment lead and aluminum levels. The intrinsic toxicity of EDTA on cells was investigated. EDTA concentrations above 600 uM reduced cellular energy metabolism. CONCLUSION: Based on our data, we proposed that low dosages of chelating agents might be preferential for patients with no– occupational exposure, as the benefit of chelation is not linear with dosage, the risk of exposure to antidotes is increasing with increased dosages, and excretion of essential metals significantly increases with increased antidote dosage. Keywords: NADH, chelation, lead, essential metals ÖZET GİRİŞ: Birçok çalışmada, metal zehirlenmelerine fizyolojik, biyokimyasal ve davranışsal fonksiyon bozukluklarının eşlik ettiği rapor edilmişitir. Toksik metallere maruz kalım tedavisi için yıllardır şelasyon kullanılmaktadır. Şimdiki klinik çalışmamızda, toksik metallere maruz kalımın tedavisinde, farklı şelasyon protokollerini ve iki EDTA formunu (sodyum kalsiyum EDTA ve sodyum EDTA) karşılaştırdık. YÖNTEM: Ca-EDTA veya Na-EDTA ile tedavi öncesinde ve sonrasında, her denekten 24 saatlik idrar örnekleri toplandı. Toksik ve esansiyel metallerin düzeyleri, grafit fırınlı atomik absorbsiyon spektrometresi (AAS) ve indüktif olarak eşleştirilmiş plazma atomik emisyon spektrometresi (ICP-AES) ile ölçüldü. Hücresel ATP düzeyleri, ATP-biyolüminesans analizi ile saptandı. Mitokondriyal potansiyel, hücrelerin mitokondri boyası ile boyanmasından sonra florimetre ve akış sitometresi ile ölçüldü. BULGULAR: Çeşitli şikayetleri olan fakat akut toksik mineral maruz kalımı olmayan, Na-EDTA ve Ca-EDTA ile şelasyon tedavisi alan 600’ün üzerindeki hastadan elde edilen veriler değerlendirildi. Ca-EDTA ve Na-EDTA’nın tedavi başına 3 g intravenöz infüzyonu, kurşunun vücuttan uzaklaştırılmasında eşit derecede etkinken; Ca-EDTA, alüminyumun uzaklaştırılmasında daha etkindi. Kurşunun uzaklaştırılması, doz bağımlıydı; fakat, doğrusal değildi. Farklı Na-EDTA dozları (3g ve 1g) ile şelasyon, idrar kurşununda %50’den daha az ortalama farklılıkla sonuçlandı. Ayrıca, esansiyel minerallerdeki eksiklikler, tedavi öncesi daha fazla kurşun ve alüminyum düzeyleri ile orantılıydı. Hücreler üzerine EDTA’nın intrensek toksisitesi araşıtırıldı. EDTA, 600 µM üzerindeki konsantrasyonlarda hücresel enerji metabolizmasını azalttı. SONUÇ: Verilere göre; şelasyonun faydası dozla orantılı olmadığından, antidotlara maruziyet riski yüksek dozlarda arttığından ve esansiyel metallerin atılımı yüksek antidot dozları ile arttığından dolayı, şelasyon ajanlarının düşük dozları mesleki olmayan maruz kalımlar için öncelikli olabilir. Anahtar Kelimeler: NADH, şelasyon, kurşun, esansiyel metaller Corresponding Author: Received March 01, 2011; accepted May 04, 2011 Nina Mikirova, DOI 10.5455/spatula.20110504045803 Riordan Clinic, Wichita, KS, USA, email: [email protected]. Published online in ScopeMed (www.scopemed.org). Spatula DD. 2011; 1(2): 81-89. Spatula DD 81 EDTA chelation therapy INTRODUCTION conducted in vitro experiments examining effects of each chelating agent on cellular lead toxicity, The build-up of heavy metals in the body can mitochondrial potential, and ATP production. lead to a variety of health problems [1-3]. Poisonous or nonessential metals such as lead (Pb), mercury MATERIALS AND METHODS (Hg), cadmium (Cd), and aluminium (Al) have been associated with pathologies in the nervous, Clinical Studies cardiovascular, haemotopoietic, gastrointestinal and Data from 600 patients treated with chelation immunological systems as well as renal dysfunction, therapy over a ten-year period (1999-2010) were anaemia, liver problems, cancer, and Alzheimer’s analyzed. Each patient exhibited long-term effects of disease [4-15]. In addition, nutritional trace elements toxic metal exposure. Prior to the onset of treatment, can be toxic at concentrations outside their each patient was screened for adequate kidney physiological range. Excessive levels of iron (Fe), functioning by assaying fasting serum creatinine manganese (Mn), copper (Cu), chromium (Cr) and (levels <1.7mg/dL were considered acceptable). zinc have been associated with conditions such as Four different chelation protocols were used over degenerative brain diseases, oxidative damage and this time period, each involving infusions in either heart problems [16-20]. Thus, it is important to sterile water or Ringers solution. 1) Na-EDTA regulate essential minerals while limiting toxic metal Normal: 3g Na-EDTA, 3 hr IV infusion, build-up in the body, and a means of clearing toxic supplements: 15g vitamin C, vitamin B6, or excess metals from the body is needed. magnesium chloride, and heparin. 2) Na-EDTA Chelation therapy has been proposed for Mini: 1g Na-EDTA, 1.5 hr IV infusion, removing poisonous metals such as Pb, Hg, Cd, and supplements: vitamin B6, and magnesium chloride. Al, as well as reducing abnormal accumulations of 3) Ca-EDTA Normal: 3g Ca-EDTA, 1 hr IV trace nutrients such as Fe, Cu, and Zn [21-24]. infusion, supplements: 15 to 25 g vitamin C, vitamin Chelation therapy employs anionic chelating agents B6, and magnesium chloride. 4) Ca-EDTA Mini: such as ethylene diaminetetraacetic acid (EDTA) to 0.10 g Ca-EDTA, 10 min IV infusion, supplements: bind heavy metal cations found in the blood. Once vitamin B6, and magnesium chloride. Regarding the EDTA bound, these metals can be removed through supplements used, vitamin B6 was used to prevent the kidneys. “chelation headaches” while magnesium chloride There is evidence that EDTA chelation benefits was used to prevent cramps associated with patients by removing toxic and trace elements from magnesium deficiency. Vitamin C is thought to diseased organs and blood vessel walls [25-29]. The assist in lead removal, and heparin was to prevent US Food and Drug administration has approved clotting at the infusion site. Patients were given ten intravenous use of sodium EDTA (Na-EDTA) for or more treatments, with mineral levels being assays treatment of hypercalcaemia, but its broader use in before and after each treatment. improving cardiovascular health is still considered Before and after the first treatment, body burden controversial [30-34]. Because Na-EDTA therapy of heavy metals, including lead, was determined by requires very slow (3 hr for a 3g IV dose) infusion, collecting urine over a twenty-four hour period for calcium EDTA (Ca-EDTA) has been employed in its analysis. Aluminium, lead, cadmium and manganese place. Ca-EDTA is FDA approved for treating levels in urine were measured by the use of Perkin- serious cases of lead poisoning. Elmer 4100ZL AAS with graphite furnace by The purpose of our study is to compare the standard protocols. Calcium, magnesium, iron, effects Na-EDTA and Ca-EDTA on toxic and copper and zinc were also analyzed on a Perkin- essential mineral levels in the body. We analyzed Elmer ICP-AES (Model 3300 DV). data collected over a ten-year period in over six- hundred patients given chelation therapy, measuring Statistics levels of Al, Pb, Cd, Mn, Ca, Mg, Fe, Cu, and Zn. Data from the various protocols were compared Over this time period two chelation regiments were statistically as follows. A Kolmogorov-Smirnov test employed for each chelating agent: a “normal” was used to determine if the data were normally regiment employing three grams of the chelating distributed. If not, Wilcoxon test was used for agent along with 15–25 grams of intravenous comparing groups. If analysis supported the normal vitamin C; and a “mini” regiment employing lower distribution of data, data were analyzed using chelating agent doses (1 g for Na-EDTA, 0.05-0.1 g Student’s t-test. P-values of 0.05 or less are for Ca-EDTA) without vitamin C. We also considered statistically significant. 82 Spatula DD