Effects of colloidal metallic silver in tablet form administered orally at 75mg/day for 2 months on various clinical and physiological parameters in six adult volunteers.
Silver belongs to the transition group I in the Periodic Table and exists in nature in two basic forms: Metallic silver which is neutral with a charge of zero ([Ag.sup.0]); and ionic silver with one positive charge ([Ag.sup.+]). The metallic form acquires new physical, chemical, and physiological properties when finely divided with particle sizes in the low nanometer range (1 to 10 nanometers). One nanometer is one billionth of a meter. Aqueous dispersion of nanocolloidal silver possesses unique characteristics, due to the fact that a significant percentage of silver atoms are exposed at the surface of the particles, capable of interacting with the aqueous medium and other molecules (1).
The metallic colloidal form and the ionic form of silver has been in use worldwide for the past 100 years as anti-germieidal agents (2). A recent review of the literature suggests that colloidal metallic silver is extremely safe whereas ionic silver is often associated with side effects (1). Since both forms of silver are biologically active and metallic silver is safer than ionic silver, one of us (GEA) decided to investigate potential clinical applications of colloidal silver not related to its antibacterial effects and to assess its toxicity when used in effective amounts.
In this manuscript, the effects of colloidal silver in tablet form on various clinical and physiological parameters in 6 subjects not taking nutritional supplementation are reported. To summarize the results of this study, colloidal silver of particle sizes less than 10nm used in daily amounts of 75mg for 2 months displayed a normalizing effect on thyroid volume and on some elemental, biochemical, and hematological parameters. The two subjects with the largest thyroid volumes responded to colloidal silver with marked reduction in the size of the thyroid. There was no significant effect of colloidal silver on the volumes of the thyroid in the other four subjects.
Colloidal silver normalized fasting blood sugar levels, decreasing it in one subject with hyperglycemia and increasing it in the other 5 subjects with hypoglycemia. It normalized red blood cells (RBC) calcium levels. One subject had elevated RBC calcium levels and the levels of the other 5 subjects values were in the upper limits of the reference range. After 1 month and 2 months on colloidal silver, RBC calcium values decreased markedly and clustered around the middle of the reference range. It improved the depression score of one of the volunteers with moderate depression, but had no significant effect on the normal scores of the other 5 subjects. There was no evidence of overt toxicity clinically nor based on hematology, blood chemistry, and urinalysis. The most significant adverse effect of colloidal silver in these 6 subjects was a marked and statistically significant decrease in Red Blood Cell and plasma copper levels.
To our knowledge, this is the first study of the metabolism of colloidal silver and its effects on parameters not related to infectious diseases. These preliminary results suggest that colloidal silver properly characterized and administered under medical supervision in amounts of 75mg/day for 2 months is safe and possesses several unexpected beneficial effects with potential clinical applications. Further research in a larger population on colloidal silver and nutritional supplementation is warranted.
II - Research Subjects
Six adult volunteers (3 men and 3 women) in overall good health, ambulatory and on no medication were recruited by one of us (J.D.F) from friends and from the local community. They ate an omnivorous diet without any restrictions and they were not on dietary supplements. The anthropometric data on these 6 subjects are displayed in Table I. the mean age was 53.3 years with a range of 25 to 72 years. The body mass index (BMI) is the ratio of body weight divided by height squared, using metric units of kilogram (kg) for weigh and meter (m) for height. The normal range is 18.5-24.9 kg/[m.sup.2], with less 18.5 as underweight; between 25-29.9 as over weight and 30 and above as obese. Based on this classification, three subjects had normal weight and the other three subjects were obese.
Table I Anthropometric Data on the 6 Study Subjects SS# Sex Age Height Weight BMI* Body Weight (years) (inches) (pounds) (Kg/[m.sub.2]) Status 1 F 50 62 127 23.2 Normal Weight 2 M 25 70 170 24.4 Normal Weight 3 F 68 66 215 34.7 Obese 4 M 72 70 244 35 Obese 5 F 50 68 155 23.6 Normal Weight 6 M 55 76 275 33.5 Obese Mean 53.3 68.7 198 29.1 S.D 18.6 3.4 49 6.1 * BMI = Body Muss Index. Normal: 18.5-24.9; Overweight: 25-29.9; Obese = 30 and over
III - Preparation and Characterization of Colloidal Silver
Colloidal metallic silver was prepared by one of us (GEA) using a proprietary procedure. USP grade silver nitrate was dissolved in purified water at a concentration of 5000 PPM (5000 mg of elemental silver per liter of water) and reduced to colloidal silver using food grade Maltodextrin as reductant and stabilizer (1), (3). The nitric acid released was neutralized to pH=7.0 [+ or -] 0.1 by USP grade potassium hydroxide. The concentration of elemental silver was measured in the liquid preparation by X-ray Fluorescence Excitation Analysis using a commercial laboratory (West Coast Analytical Services, Inc., Santa FE Springs, CA). In various batches, the levels ranged from 4850 PPM to 5300 PPM.
Particle size analysis by Transmission Electron Microscopy (TEM) was performed on the liquid preparations, courtesy of Ralph Albrecht and Joe Heintz at The University of Wisconsin-Madison, Madison WI. The histogram of particle distribution of the batch used in the present study is displayed in Fig I. The diameter of the colloid particles ranged from 3nm to 10nm (1).
[FIGURE 1 OMITTED]
Since ionic silver is toxic and colloidal metallic silver is safe (1), care was taken to ensure that all the ionic silver nitrate was reduced to metallic colloidal silver. A silver ion selective electrode (Orion P.N. # 9616BNWP) was used in pilot studies to measure ionic silver during the reduction of silver nitrate until the ionic silver became undetectable (less than 0.5 PPM). The ion selective silver electrode does not detect metallic neutral silver. It was arbitrarily decided that an acceptable ratio of total silver/ionic silver for uses in human subjects should be 5000 or greater (3), which gave a wide margin of safety. Since the concentration of elemental silver averaged 5000 PPM in the colloid suspensions and ionic silver concentrations were less than 0.5 PPM, the total silver/ionic silver ratio was greater than 10,000.
The high ratio of total silver/ionic silver was confirmed by measuring silver concentration in the supernatant of one of the batches following ultra centrifugation (3). This procedure precipitates colloidal silver particles 1nm and larger; and leaves only ionic silver in the supernatant. The concentration of silver in the supernatant was 0.07 PPM, giving a ratio of total silver/ionic silver over 50,000.
The aqueous suspension of colloidal silver was evaporated to less than 3% moisture and compounded with a silica excipient (micosolle[R]) to achieve a concentration of 12.5mg silver/tablet. From the elemental analysis of the silver colloid powder by ICPMS, all the toxic metals were below the sensitivity of the assay: As <0.001ppm; Se <0.001ppm; Cd <0.001ppm; Sb <0.001ppm; Hg <0.0001ppm; and Pb <0.001ppm. A bacteriostatic agent was not required in the liquid colloidal silver preparation since colloidal silver has itself germicidal properties. The colloidal silver tablets were packaged in 90 tablets/bottle.
IV - Research Protocol
This study was performed at the Flechas Family Practice (FFP) Clinic in Hendersoville, NC and supported by a grant from Optimox Corporation. After informed consent, the subjects underwent a complete history and physical examination prior to intervention and after 2 months on colloidal silver. After initial evaluation, each subject was supplied with four bottles of colloidal silver tablets containing 90 tablets with 12.5mg silver per tablet and instructed to ingest 6 tablets a day for 8 weeks and to report any adverse effects. The tests performed on these subjects are listed in Table II.
Table II Self-Administered Questionnaires, Clinical Evaluation, and Laboratory tests Performed on the 6 Subjects Procedure & Tests Facility Pre-Intervention Post 1 Performed Where Tests Month Were on Silver Performed Physical Exam + FFP X Vital Signs Body Composition FFP X X Basal Metabolic Rate FFP X X Self Administered FFP X X Symptom Questionnaire Zung's Depression Scale FFP X X Urinalysis FFP X X Thyroid Ultrasonometry FFP X CBC Lab Corp X X Blood Chemistry Lab Corp X X Antinuclear Antibodies Lab Corp X (ANA) Thyroid Function Lab Corp X Tests + TPO Ab RBC Glutathione Genova X X Peroxydase Diagnostics Urine Lipid Peroxides Genova X X Diagnostics Red Cell Elemental Doctor's Data X X Analysis Including Silver by ICP-MS Measurement Doctor's Data X X of Silver & Copper in Plasma, Urine and Saliva by ICP MS Procedure & Tests Post 2 Performed Month on Silver Physical Exam + X Vital Signs Body Composition X Basal Metabolic Rate X Self Administered X Symptom Questionnaire Zung's Depression Scale X Urinalysis X Thyroid Ultrasonometry X CBC X Blood Chemistry X Antinuclear Antibodies X (ANA) Thyroid Function X Tests + TPO Ab RBC Glutathione X Peroxydase Urine Lipid Peroxides X Red Cell Elemental X Analysis Including Silver by ICP-MS Measurement X of Silver & Copper in Plasma, Urine and Saliva by ICP MS
The daily amount of 75mg colloidal silver was chosen because of our previous experience with daily amount of 12.5 to 75mg of colloidal silver in tablet form. When tested in normal subjects on a complete dietary supplement program for up to 2 years, six tablets/day (75mg) gave the best results for mental, physical and overall wellbeing; physical energy, mental alertness; clearing of sinus infection, resistance to cold and flu; visual acuity for both close and distant vision. Some subjects with constipation reported improved bowel movement with 6 tablets/day but not with lesser amounts. One female subject who experienced thick foul smelling vaginal discharge following antibiotics for oral surgery, was told to ingest 2 tablets of colloidal silver a day for one week, then 4 tablets/day for one week; and 6 tablets/day for one week. She noticed no effect on 2 tablets/day. There was a decreased vaginal discharge on 4 tablets/day. After 3 days at 6 tablets/day, the vaginal discharge cleared completely. She stopped taking the colloidal silver after one week at 6 tablets/day. Three months later, she was still asymptomatic. Another female subject experienced repeated bouts of upper respiratory tract infections treated with antibiotics for several years. Each episode of antibiotics was associated with vaginal discharge and vaginal moniliasis. Some 18 months ago, she was placed on a complete nutritional program together with 6 tablets of colloidal silver/day. The infection and discharge cleared after 3 days on the colloidal silver and she has not experienced a recurrence since.
In these subjects, on nutritional supplementation, even after 2 years on colloidal silver, there was no evidence of argyria (blue grayish discoloration of the teguments), or other significant side effects. Hematology, Blood Chemistry and Urine Analysis pre and post colloidal silver showed no evidence of toxicity. Based on these preliminary results, the present study was designed to investigate the beneficial and side effects of colloidal metallic silver at 6 tablets/day in a small group of subjects not taking dietary supplements who were followed with a more extensive evaluation of possible toxicity and physiological effects. The following clinical evaluations and tests were performed prior to intervention; after 4 weeks and 8 weeks on silver colloids of 75mg/day. Paired data statistic was used to compare data values obtained before intervention with values obtained on the same parameters following colloidal silver at 75mg/day for 1 month and 2 months.
* Two questionnaires were completed by the subjects before intervention, post one month and 2 months of intervention: the Symptomatology Questionnaire and the Zung Depression Scale. The self administered Symptom Questionnaire included several mental, emotional and physical symptoms and was composed by the authors to assess possible toxicity and beneficial effects of colloidal silver used at 75mg/day for 2 months. It contains 33 items, 28 symptoms and 5 items assessing overall performance. The scoring of the Symptomatology Questionnaire used an analog scale from 1 to 10, 1 being the worse and 10 being the best (no symptom). The Zung's Depression Scale is a measure of the severity of depression with score below 50 as within the reference range; of 50-59 as mild to moderate depression, 60-69 as moderate to marked depression; and greater than 70 as severe depression (4).
* Urine analysis was performed at the clinic with Multistix 10SG reagent strips and read on a Clinilek 100 that was calibrated daily.
* Complete blood count (CBC), the metabolic panel, thyroid profile, TPO antibody titer and ANA titer were performed by Lab Corporation of America.
* Thyroid ultrasonometry was computed by a registered sonographer using a portable Biosound Esaote Megas System unit with a frequency of 7.5MHz
* Red Blood Cells (RBC) elemental analysis was performed at a commercial laboratory, which supplied the kits for sample collection. Heparinized blood was collected before intervention one month and 2 months post-colloidal silver. After centrifugation packed red blood cells (RBC) were separated from plasma and mailed to Doctor's Data, St. Charles, IL.
The complete panels of essential plus toxic elements were measured ICPMS in the RBC; silver and copper levels were measured also in the separated plasma in 24hr urine collections and saliva samples collected prior, 1 month and 2 months post colloidal silver.
* Genova Diagnostics supplied the kits for the measurement of red blood cell glutathione peroxydase and urine lipid peroxides
* The Bio Impedance Analyzer, Model BIA-450 from Biodynamics, Seattle, WA was purchased by Optimox Corporation and made available to the FFP clinic for measurement of Basal Metabolic Rate (BMR) and body composition.
A--Side effects and beneficial effects reported by the subjects.
Clinical evaluation during follow-up revealed that 2 subjects experienced side effects and 2 subjects reported beneficial effects. We will discuss first the two subjects with side effects. Subject #2 experienced muscle pain in the shoulder and neck at the beginning of the study. The pain lasted one week and subsided afterward. Subject #5 experienced muscle pain in her shoulder and neck one week after starting the colloidal silver. The pain persisted throughout the whole 2 month study period and lasted up to 3 weeks after the end of the study. She will be discussed further in the results and discussion section.
The beneficial effects observed in the 2 subjects are as follows. Subject #3 had a long history of fungal infection between the fingers not responding to various remedies. After one month on colloidal silver at 75mg/day, the fungal infection disappeared and she has been symptom free since. Subject #1 suffered from fibromyalgia controlled with iodine tablets (Iodoral[R]) combined with vitamins B-2 and B-3 (ATP Cefaclors). When she stopped the iodine supplementation, her muscle pain recurred within a day or two. She did not take the iodine/B vitamins supplements during the first 6 weeks of this study. Her fibromyalgia symptoms did not recur while on colloidal silver, even though she was not taking the iodine--B-vitamin supplements. Subject #1 scored high at 63 on Zung's Depression Scale to a level of moderate to marked depression prior to colloidal silver. From a score of 63, her score decreased to 49 after 2 months on colloidal silver. Less than 50 is considered within the normal range.
B--Itemized effects of colloidal silver.
1. Clinical Parameters
There was no significant change in systolic and diastolic blood pressures and body temperature (data not shown). Basal metabolic rate, lean body mass, fat mass, total body water, extra cellular and intracellular water are displayed in table III. The mean body weight decreased by 5 lbs after 2 months on colloid silver (p = 0.23). Mean fat mass decreased also by 5 lbs (p = 0.18) post intervention. There was no significant effect of colloidal silver on body weight, Basal Metabolic Rate, lean body mass, fat mass, total body water, intracellular and extracellular water (Table III).
Table III Effects of Colloidal Silver Administered Orally at 75mg/day for 2 Months on Body Weight, Body Composition, and Basal Metabolic Rate (BMR) in 6 Volunteers. Pre mean Post mean p Value [+ or -]SD [+ or -]S.D. Body Weight (lbs) 198[+ or -]47 193[+ or -]61 (p = 0.2) Basal Metabolic Rate 1905[+ or -]476 1926[+ or -]497 (p = 0.23) (cal/day) Fat Mass (lbs) 63[+ or -]27 57.8[+ or -]32 (p = 0.40) Lean Mass (lbs) 135[+ or -]26 136[+ or -]32 (p = 0.48) Total Body 19[+ or -]6.0 18.2[+ or -]5.3 (p = 0.28) Water (liters) Intracellular 25.5[+ or -]4.9 22.9[+ or -]7.9 (p = 0.4) Water (Liters) Extracellular Water 19[+ or -]6.1 18.2[+ or -]5.3 (p = 0.22) (Liters)
Of the 28 symptoms assessed in the Symptomatology Questionnaire, none showed significant change in the mean scores between baseline and 2 months post silver at p<0.05. The scoring of mostly 8's, 9's and 10's before silver supplementation suggest that the study subjects, although not in perfect health were overall in good physical and mental conditions. The scores of 8's for job performance, relationship with others, physical, mental and overall well being concur with the high scores of the 28 symptoms assessed. If we consider/? values between 0.05 and 0.1 as near significant, four of the symptoms improved following colloid silver: brain fog (p=0.08), constipation (p=0.1), forgetfulness (p=0.09) and resistance to flu/cold (p=0.1). Physical well being, mental well being and overall well being improved following colloidal silver to near significance (Table IV).
Table IV Effects of Colloidal Silver at 75mg/day for two months in six volunteers on their scoring of the Symptomatology Questionnaire, using an analog scale from 1= severe to 10 = no symptom best possible. Symptoms Pre Silver mean S.D. Post Silver mean S.D. Muscle Pain 7.8[+ or -]3.1 7.7[+ or -]2.6 Joint Pain 7.8[+ or -]2.3 8[+ or -]1.4 Joint Swelling 9[+ or -]2.0 8.5[+ or -]2.1 Pain on Exercise 8[+ or -]1.9 7[+ or -]3.1 Restless Legs 8.2[+ or -]1.6 7.3[+ or -]2.3 Leg Cramps 8.2[+ or -]1.7 6.8[+ or -]2.8 Stiffness 7.3[+ or -]2 7.3[+ or -]3.0 Fatigue 7.5[+ or -]2.1 7.8[+ or -]1.7 Irritability 8.5[+ or -]1.1 8.2[+ or -]1.2 Anxiety 8.7[+ or -]2.8 9.3[+ or -]0.5 Depression 8.3[+ or -]1.6 8.9[+ or -]2.0 Dizziness 9.7[+ or -]0.8 9.7[+ or -]0.8 Haziness 9[+ or -]1.1 8.5[+ or -]3.2 Insomnia 8.8[+ or -]1.6 7.8[+ or -]3.5 Brain Fog 8.2[+ or -]1.8 9[+ or -]1.1 Panic Attack 9.3[+ or -]1.0 9.5[+ or -]0.8 Frequent Urination 8.5[+ or -]1.8 8.2[+ or -]2.2 Fever 10[+ or -]0.0 10[+ or -]0.0 Flushing 8.8[+ or -]2.4 8.8[+ or -]2.0 Nasal Congestion 7.7[+ or -]1.8 6.2[+ or -]1.7 Abdominal Cramping 9.5[+ or -].22 8.5[+ or -]3.2 Burning on Urination 9.7[+ or -]0.5 9.5[+ or -]0.8 Constipation 9.3[+ or -]1.0 9.8[+ or -]0.4 Hostility 9.3[+ or -]0.5 8.8[+ or -]1.2 Diarrhea 9[+ or -]1.3 8.2[+ or -]3.5 Forgetfulness 7.8[+ or -]2.5 9[+ or -]3.5 Resistance to Flu/Cold 7.3[+ or -]1.8 9.2[+ or -]0.8 Coordination 9.3[+ or -]0.5 8.2[+ or -]0.4 Equilibrium 9.3[+ or -]0.5 9.3[+ or -]0.5 Sinus Drainage 7.8[+ or -]1.8 6.6[+ or -]1.9 Motivation 7.5[+ or -]2.4 8.5[+ or -]1.1 Visual Near 7.8[+ or -]1.8 8.5[+ or -]1.2 Visual Distant 6.7[+ or -]3.1 7.3[+ or -]3.1 Job Performance 8[+ or -]2.5 8.3[+ or -]1.8 Relationship with Other 8.8[+ or -]0.8 8.7[+ or -]1.03 Physical Wellbeing 8.2[+ or -]3.1 9.3[+ or -]0.8 Mental Wellbeing 8.3[+ or -]2.7 9.3[+ or -]0.8 Overall Wellbeing 8.2[+ or -]3.1 9.3[+ or -]0.8 Symptoms p Value Muscle Pain 0.46 Joint Pain 0.37 Joint Swelling 0.24 Pain on Exercise 0.14 Restless Legs 0.18 Leg Cramps 0.14 Stiffness 0.48 Fatigue 0.32 Irritability 0.18 Anxiety 0.28 Depression 0.26 Dizziness 0.50 Haziness 0.31 Insomnia 0.18 Brain Fog 0.10* Panic Attack 0.18 Frequent Urination 0.18 Fever 0.50 Flushing 0.50 Nasal Congestion 0.21 Abdominal Cramping 0.14 Burning on Urination 0.18 Constipation 0.10* Hostility 0.21 Diarrhea 0.24 Forgetfulness 0.09* Resistance to Flu/Cold 0.10* Coordination 0.18 Equilibrium 0.50 Sinus Drainage 0.24 Motivation 0.15 Visual Near 0.18 Visual Distant 0.18 Job Performance 0.18 Relationship with Other 0.18 Physical Wellbeing 0.09* Mental Wellbeing 0.08* Overall Wellbeing 0.08* *= Near significant improvement: 0.05 < p=0.1
Zung Depression Scale index dropped from a preintervention mean value [+ or -] SD of 39 [+ or -] 12.8 to a post 2 months silver supplementation of 36.8 [+ or -] 8.2, which was not significant at p<0.05 (Table V). Except for subject #1, the scores were within the normal range for the remaining 5 subjects, pre and post intervention and no significant effect of the colloidal silver on these normal values were observed. However, subject #1 with the highest score of 63 dropped to 49 after 2 months on the silver preparation. A Zung score between 60 and 69 is considered indicative of moderate to marked depression, and below 50 as within the normal range of values (4). Of interest is the fact that subject scored the lowest of the 6 subjects for anxiety and depression in the Symptomatology Questionnaire prior to colloidal silver, with score of 3 for anxiety and 6 for depression. These scores increased to 7 and 8 after 1 month on colloidal silver; 9 and 7.5 after 2 months on colloidal silver. An increase in this scoring System means an improvement of symptoms.
Table V The Zung Depression Scores* in 6 volunteers prior to intervention and following 2 months on colloidal silver at 75mg/day SS# Pre Silver Post Silver 1 63 49 2 35 33 3 25 37 4 39 44 5 34 29 6 38 29 Mean 39 36.8 S.D 2.8 8.2 p Value 0.59 --
2. Thyroid Gland and Thyroid Hormones
Ingestion of colloidal silver at 75mg/day resulted in a selective reduction in the thyroid volume of two of the subjects with the largest thyroid size (15 and 22.8 [cm.sup.3]) but no significant effect was observed in the other four subjects with thyroid volumes ranging from 7.8 [cm.sup.3] to 10.9 [cm.sup.3] (Table VI). As a group, the mean thyroid volume decreased from 12.8[+ or -]5.5 [cm.sup.3] to 10.5[+ or -]2.0 after 2 months on colloidal silver with p value of 0.08. This decrease in the mean value was mainly due to the marked reduction in thyroid volume of subjects #4 and #6.
Table VI Effects of Colloidal Silver at 75mg/day for 2 Months on Thyroid Volume, thyroid cysts and masses in 6 Volunteers. Thyroid Volume (cm3) Cyst/Mass SS# Pre Post Pre Post 1 10.5 11.3 None None 2 7.8 10.1 None None 3 10.9 7.7 Rt Cyst Rt Cyst 2x3cm 2x3cm 4 15 8.8 Rt Mass Rt Mass 0.7x1.0 Lt 0.8x1.1 Lt Mass 0.6x0.9 Mass 0.7x0.9 5 9.7 9 Rt Cyst Rt Cyst 2x3cm 2x3cm 6 22.8 15.9 Lt mass Lt mass 0.8x0.9cm 1x1.4cm Mean 12.8 10.5 -- -- SD 5.5 2.0 -- -- p Value -- 0.08 -- --
There was ultrasound evidence in subjects #4 and #6 of masses with pre silver sizes of 0.8cm x 0.9 cm in the left lobe of subject #6; and with masses in both lobes in subject #4, 0.7 x 1.0 cm on the right side and 0.6 x 0.9 cm on the left lobe. All 3 masses in the 2 subjects increased in size following 2 months on colloid silver (Table VI). In subject # 4 the right mass increased from 0.7 x 0.9 cm to 0.8 x 1.1 cm, and the mass in the left lobe increases from 0.5 x 0.9 cm to 0.7 x 0.9cm. In subject #6, the left mass increased from 0.8 x 0.9 cm to 1 x 1.4 cm. Subject #6 was referred to his primary care physician for a fine needle biopsy and follow up. Subject #4 and #6 were asked to continue on the colloidal silver. A repeat ultrasound by the radiologist one month later while subject #6 was still on colloidal silver for a total of 3 months did not show the nodule which had disappeared.
We repeated the ultrasound analysis 4 months after the beginning of the study. Subject #4 at that time was taking colloidal silver for 31/2 months and subject #6 for 4 months. The results of the repeated ultrasound at 4 months post silver in subject #4 and #6 are combined with the previous data on table VII for case of comparison. In subject #4, the volume of thyroid gland decreased from 15[cm.sup.3] pre-silver to 8.8[cm.sup.3] after 2 months and 8.1cm3 after 31/2 months on colloidal silver. At that last evaluation, the right mass decreased to 0.4x0.5[cm.sup.3] and the left mass to 0.2x0.3[cm.sup.3]. In subject #6, a normal echo-texture without any mass was reported at 4 months post colloidal silver. The volume of the thyroid decreased from 22.8[cm.sup.3] pre-intervention to 15.9[cm.sup.3] after 2 months and 16.5[cm.sup.3] after 4 months on colloidal silver.
Table VII Effects of Colloidal Silver at 75mg/day for 4 Months on Thyroid Volume, and Masses in Subjects #4 and #6. SS# Pre-Colloidal Silver At 2 Months Thyroid Volume 15cm3 8.8cm3 Subject 4 Masses Right 0.7x1.0cm 0.8x1.1cm Left 0.6x0.9cm 0.7x0.9cm Subject 6 Thyroid Volume 22.8 cm3 15.9 cm3 Masses Left 0.8x0.9cm 1x1.4cm SS# At 4 Months Thyroid Volume 8.1 cm3 Subject 4 Masses 0.4x0.5cm 0.2x0.3cm Subject 6 Thyroid Volume 16.5 cm3 Masses No Mass Normal Echo-Texture
The serum levels of TSH, [T.sub.3], [T.sub.4], Tree [T.sub.3] and Free [T.sub.4] were within the reference range for all the subjects prior to intervention and remained within the reference range 2 months post colloidal silver at 75mg/day, (Table VIII). There was a near significant increase (p = 0.07) of the mean TSH level post colloidal silver (2.8 [+ or -] 1.4) compared to pre intervention (2.05 [+ or -] 1.1). No significant change was observed in the thyroid hormone levels following 2 months on colloidal silver.
Table VIII Effects of Colloidal Silver Administered Orally at 75mg/day on Thyroid Function Tests in 6 Volunteers Test (u IU/L) Pre mean[+ or -]SD Post mean[+ or -]SD p Value TSH 2.05[+ or -]1.1 2.8[+ or -]1.4 0.07 Total T4 (ug/dl) 7.7[+ or -]0.9 7.6[+ or -]0.87 p = 0.38 Total T3 (ng/dl) 129[+ or -]6 120[+ or -]4.3 p = 0.33 Free T4 (ng/dl) 1.2[+ or -]0.09 1.13[+ or -]0.05 p = 0.20 Free T3 (pg/ml) 3.3[+ or -]0.23 3.3[+ or -]0.25 p = 0.32
Urine analysis results were normal in all 6 subject pre intervention and 2 months following supplementation with colloidal silver. There was no significant difference between pre and post intervention. (Data not shown)
There was a significant effect of colloidal silver on several hematological parameters (Table IX). Red blood cell count decreased from a mean of 4.92 [+ or -] 0.28 x [10.sup.6]/mL pre intervention to a mean of 4.73 [+ or -] 0.21 [10.sup.6]/mL after 2 months on colloidal silver (p <0.05), that is a 4% drop. Hematocrit mean values were significantly lower post intervention with means [+ or -] SD of 45.5 [+ or -] 2.7 before colloidal silver and 43.3 [+ or -] 2.9 after 2 months on colloidal silver (p <0.05) representing a 5% decrease. The mean value of [+ or -] SD of hemoglobin concentrations before colloidal silver was 14.6 [+ or -] 1.1, compared to a mean of 14.1 [+ or -] 1.1 post silver a near significant drop (p = 0.05) of 3.4%. Red blood cell distribution width (RDW) decreased (p = 0.07) following colloidal silver, from pre silver mean [+ or -] SD of 14.7 [+ or -] 1.1 to 14.3 [+ or -] 1.1.
Table IX Effects of Colloidal Silver at 75mg/day for 2 months on hematology in 6 volunteers Units Reference Range Pre-Silver mean[+ or -]SD WBC 10 3/uL 4.6-10.2 5.60[+ or -]2.0 RBC 10 6/uL 3.8-6.5 4.92[+ or -]0.28 Hb g/dl 11.5-18 14.57[+ or -]1.10 Hct % 37-54 45.50[+ or -]2.68 MCV fL 80-100 92.67[+ or -]4.80 MCH Pg 27-32 29.65[+ or -]1.88 MCHC g/dL 31-36 32.00[+ or -]0.66 RWD % 11.5-15 14.70[+ or -]1.12 Plat 10 3/uL 150-400 221.83[+ or -]67.71 Post-Silver mean[+ or -]SD p Value WBC 5.87[+ or -]1.7 0.25 RBC 4.73[+ or -]0.21 <0.05 Hb 14.13[+ or -]1.07 0.05 Hct 43.33[+ or -]2.92 <0.05 MCV 91.87[+ or -]4.80 0.15 MCH 29.87[+ or -]1.83 0.19 MCHC 32.62[+ or -]0.69 0.2 RWD 14.32[+ or -]1.06 0.07 Plat 229.17[+ or -]58.04 0.30
5. Blood Chemistry
The data on the metabolic panel (blood chemistry) arc displayed in Table X. Fasting blood glucose levels as a whole increased from 66.6 [+ or -] 29 to 75 [+ or -] 21 with a p value of 0.16. However, when the data on blood glucose levels are analyzed individually (Table XI), it reveals that subject #4 with hyperglycemia experienced a drop from 125mg/dL before to 113mg/dL after 2 months on colloidal silver whereas the other 5 subjects with hypoglycemia experienced an increase in fasting blood sugar. The mean [+ or -] SD for these 5 hypoglycemic subjects was 54 [+ or -] 4.8 pre silver to 67 [+ or -] 10 post silver with p value of <0.05. Four of the five hypoglycemic subjects reached blood sugar levels within the reference range following 2 months on colloidal silver at 75mg/day.
Table X Effect of colloidal silver at 75mg/day for 2 months on blood chemistry in 6 volunteers Units Reference Range Pre-Silver mean[+ or -]SD Glu mg/dL 65-109 66.6[+ or -]29.1 BUN mg/dL 5-26 14.7[+ or -]2.14 Creat mg/dL 0.5-1.5 0.89[+ or -]0.15 BUN/Creat - - 16.3[+ or -]3.56 Na mMole/L 135-148 141.8[+ or -]1.72 K mMole/L 3.5-5.5 3.7[+ or -]0.31 CI mMole/L 96-109 99.5[+ or -]1.05 CO2 mMole/L 20-32 23[+ or -]1.25 CA mg/dL 8.5-10.6 9.5[+ or -]0.38 Prot gm/L 6-8.5 7.5[+ or -]0.69 Alb gm/L 3.5-5.5 4.65[+ or -]0.50 Glob gm/L 1.5-4.5 2.9[+ or -]0.57 A/G ratio - 1.1-205 1.8[+ or -]0.37 Total Bil mg/dL 0.1-1.2 0.5[+ or -]0.21 Alk Phos I.U/L 25-165 88.3[+ or -]27.69 SGOT I.U/L 0-40 24.8[+ or -]8.59 SGPT I.U/L 0-40 29.5[+ or -]11.76 Post-Silver mean[+ or -]SD p Value Glu 75[+ or -]20.70 0.16 BUN 19.3[+ or -]3.9 <0.01 Creat 0.87[+ or -]0.13 0.36 BUN/Creat 22.5[+ or -]3.9 <0.05 Na 141.8[+ or -]1.72 0.50 K 4.1[+ or -]0.46 <0.01 CI 99.8[+ or -]1.21 0.39 CO2 21.5[+ or -]0.84 <0.01 CA 9.4[+ or -]0.45 0.28 Prot 7.4[+ or -]0.59 0.28 Alb 4.7[+ or -]0.39 0.39 Glob 2.7[+ or -]0.38 0.26 A/G ratio 1.8[+ or -]0.25 0.50 Total Bil 0.4[+ or -]0.12 <0.05 Alk Phos 103.2[+ or -]35.36 <0.01 SGOT 28.8[+ or -]4.46 0.07 SGPT 30.5[+ or -]13.61 0.42 Table XI Fasting serum glucose* pre and 2 months post colloidal silver. Statistical Significance was achieved by excluding subject 4 with hyperglycemia. Fasting Serum Glucose (mg/dL) SS# Pre Silver Post Silver 1 54 77 2 57 65 3 61 72 4 125 113 5 51 72 6 49 51 Mean 67 75 SD 29 21 p Value = 0.155 Excluding Subject # 4 Mean 54 67 SD 4.8 10 p Value =<0.05 * Reference Range = 65-99mg/dL
Blood Urea Nitrogen (BUN) levels were significantly (p<0.01) elevated following colloidal silver with mean [+ or -] SD of 14.7 [+ or -] 2.1 mg/dL before silver administration and 19.3 [+ or -] 3.9 post 2 months on colloidal silver. However, serum creatinine levels were not affected, with values of 0.89 [+ or -] 0.15 pre silver and 0.87 [+ or -] 0.13 mg/dL post silver, resulting in a significant (p <0.05) increase in the BUN/creatinine ratio, being respectively 16.3 [+ or -] 3.6 before and 22.5 [+ or -] 3.9 after 2 months on colloidal silver. Serum potassium levels increased significantly (p<0.01) from 3.7 [+ or -] 0.31 before; and 4.1 [+ or -] 0.46 after colloidal silver. There was a significant decrease in serum [CO.sub.2] and total bilirubin levels; for [CO.sub.2] pre= 23 [+ or -] 1.25 and post = 21.5 [+ or -] 0.84 (p<0.01); For total bilirubin 0.5 [+ or -] 0.21 for pre silver and 0.4 [+ or -] 0.12 (p<0.05) for post colloidal silver. The enzymes alkaline phosphates and SGOT increased following intervention, being significant (p<0.01) for alkaline phosphates (pre = 88.3 [+ or -] 22.7 and post = 103 [+ or -] 35.4) and near significant (p=0.07) for SGOT (24.8 [+ or -] 8.6 before; and 28.8 [+ or -] 4.5 after).
6. RBC Elemental Analysis
The results of the red blood cell (RBC) elemental analysis by ICP-MS are displayed in Table XII. There was a marked drop in RBC calcium levels from 27.7 [+ or -] 4.3 to 18.5 [+ or -] 4.8 (p<0.01) post 2 months on silver. It is of interest that pre-silver RBC calcium values were in the upper limit of the reference range in 5 subjects and elevated above that range in subject #1. Following one month and 2 months on colloidal silver, the RBC calcium mean levels decreased significantly and clustered around the midpoint of the reference range, suggesting a normalizing effect of colloidal silver on RBC calcium (Fig 2).
Table XII Effect of colloidal silver at 75mg/day for 2 months on red blood cell elemental analysis in 6 volunteers. Units Reference Range Pre-Silver mean[+ or -]SD Calcium PPM 8-31 27.7[+ or -]4.32 Magnesium PPM 36-64 46.5[+ or -]3.83 Potassium PPM 65-95 73[+ or -]3.69 Phosphorus PPM 480-745 514[+ or -]51.08 Copper PPM 0.52-0.89 0.7[+ or -]0.08 Zinc PPM 8-14.5 9.9[+ or -]0.73 Iron PPM 745-1050 747.8[+ or -]57.48 Manganese PPM 0.007-0. 030 0.015[+ or -]0.00 Chromium PPM 0.0003-0.0060 0.00068[+ or -]0.00 Selenium PPM 0.19-0.38 0.208[+ or -]0.02 Boron PPM 0.01-0.110 0.024[+ or -]0.01 Post-Silver mean[+ or -]SD p Value Calcium 18.5[+ or -]4.76 <0.01 Magnesium 47.2[+ or -]2.93 0.32 Potassium 76.2[+ or -]5.56 0.17 Phosphorus 543.7[+ or -]43.20 0.20 Copper 0.57[+ or -]0.08 <0.05 Zinc 10.85[+ or -]1.42 0.11 Iron 809[+ or -]45.96 <0.01 Manganese 0.02[+ or -]0.00 0.07 Chromium 0.00083[+ or -]0.00 <0.05 Selenium 0.225[+ or -]0.03 0.05 Boron 0.033[+ or -]0.01 0.06
[FIGURE 2 OMITTED]
RBC copper levels decreased significantly post silver (p<0.05) from 0.7 [+ or -] 0.08 to 0.57 [+ or -] 0.08 PPM. There was a significant increase in the following elements: Iron (p< 0.05) from 747.8 [+ or -] 57.5 to 809 [+ or -] 46; Chromium (p<0.05) from mean values of 0.00068 to 0.00083 PPM. The mean value of following elements increased post silver to reach near significance: Selenium (p=0.05) from 0.208 to 0.225; Boron (p=0.06) from 0.024 to 0.033 PPM; Manganese (p=0.07) from 0.015 to 0.02 PPM.
The RBC levels of the toxic elements arsenic, cadmium, lead and mercury were within the reference range of the laboratory pre and post silver. There was no significant difference at p<0.05 between basal levels and post silver levels. However, RBC lead levels decreased from 0.025 [+ or -] 002 before intervention to 0.021 [+ or -] 0.0002 post silver, reaching near significance at p=0.10
7. Oxidative Burden and Autoimmunity
Red blood cell glutathione peroxydase levels and urine lipid peroxides were within the normal range before intervention and remained within the normal range following 2 months on colloidal silver (Table XIII). The mean values were respectively for pre and post intervention: for glutathione peroxydase 29.5 [+ or -] 3.2 and 28.5 [+ or -] 3.75; for urine lipid peroxides 5.7 [+ or -] 2.6 and 6.02 [+ or -] 1.8. The reference range for glutathione peroxydase is from 20 to 38 units/gin of protein; for urine lipid peroxide, it is less than 10 micromole/gm creatinine.
Table XIII The Effects of Colloidal Silver at 75mg/day for 2 Months on Red Blood Cell Glutathione Peroxydase and Urine Lipid Peroxides Levels in 6 Adult Volunteers. RBC Glutathione Peroxydase (units/gm protein) SS# Pre Post - 2 Months 1 32.6 29.5 2 30.8 29.9 3 29.2 26.7 4 28.9 26.7 5 23.7 23.5 6 31.6 34.5 Mean 29.5 28.5 SD 3.2 3.8 P Value -- 0.31 Reference = 20-38 Range Urine Lipid Peroxides (micromoles/gm creatine) SS# Pre Post--2 Months 1 6.3 7.1 2 5.3 4.1 3 7.0 5.0 4 9.6 5.5 5 3.6 9.2 6 2.2 5.2 Mean 5.7 6.2 SD 2.6 1.8 P Value -- 0.82 Reference Range<10
Anti NuclearAntibodies (ANA) were negative for all 6 subjects' pre and post 2 months on colloidal silver. The ANA test was added to the laboratory evaluation because of a 2003 publication reporting that in susceptible mice, ionic silver salts (chloride and nitrate) induced antinucleolar antibody production (5).
8. Metabolism of Silver and Copper
The concentration of silver in Red Blood Cells (RBC), plasma, urine, and saliva are displayed in Table XIV. Supplementation with colloidal silver at 75mg/day increased significantly silver levels in RBC and plasma to levels of 162.2ug/L and 161ug/L; Urine silver levels pre-supplementation were below 0.lug/L in 5 of the 6 subjects. Urine silver levels increased (p<0.05) from a mean of less than 0.13ug/L to l.5ug/L. Saliva silver levels were below 0.1ug/L in 5 of 6 subjects. From a mean of <0.22ug/L pre-supplementation the values increased to 4.9ug/L, mainly due to marked increase in subject #5. This post-silver increase in saliva silver levels was not statistically significant. Silver supplementation decreased significantly copper levels in RBC and plasma (Table XV). The effect of colloidal silver was greater on plasma copper levels (drop of 45%) than on RBC levels (drop of 15%). No significant effect of colloidal silver was observed on the levels of copper in urine. Except for subject #5 with saliva copper levels of <0.1ug/L before and 27.8ug/L after 2 months on colloidal silver, there was no significant effect of colloidal silver on saliva copper levels in the other 5 subjects.
Table XIV Silver Levels* in Red Blood Cells (RBC), Plasma, Urine and Saliva of 6 Volunteers Pre-Intervention and After 2 Months on Colloidal Silver at 75mg/Day. RBC (PPB) Plasma (PPB) SS# Pre Post--2 Months Pre Post--2 Months 1 0.2 192 0.34 142 2 <0.1 105 <0.1 111 3 <0.1 177 <0.1 196 4 0.12 279 0.6 151 5 <0.1 132 0.24 215 6 <0.1 88 0.44 149 Mean <0.12 162.2 <0.33 161 SD -- 70 -- 38 p Value <0.001 <0.001 Urine (PPB) Saliva (PPB) SS# Pre Post--2 Months Pre Post--2 Months 1 <0.1 0.55 <0.1 0.29 2 <0.1 4.7 <0.1 <0.1 3 <0.1 0.93 <0.1 0.28 4 0.28 0.42 <0.1 0.42 5 <0.1 1.3 <0.1 27.8 6 <0.1 1.3 0.82 0.65 Mean <0.13 1.5 <0.22 4.9 SD -- 1.6 -- 11.2 P Value <0.05 p>0.1 * No reference ranges available Table XV Copper Levels in Red Blood Cells (RBC), Plasma, Urine and Saliva of 6 Volunteers Pre-Intervention and After 2 Months on Colloidal Silver at 75mg/Day. RBC (PPM) SS# Pre Post--2 Months 1 0.67 0.51 2 0.57 0.53 3 0.77 0.59 4 0.6 0.58 5 0.65 0.5 6 0.75 0.72 Mean 0.67 0.57 SD 0.08 0.08 p Value <0.05 Reference Range 0.52-0.89 Plasma (PPM) SS# Pre Post--2 Months 1 1.03 0.38 2 0.69 0.47 3 1.19 0.55 4 0.96 0.56 5 1.05 0.59 6 0.97 0.73 Mean 0.98 0.55 SD 0.16 0.12 p Value <0.005 Reference Range Male 0.7-1.4 Female 0.8-1.5 Urine (PPM) SS# Pre Post--2 Months 1 0.026 0.019 2 0.055 0.053 3 0.026 0.029 4 0.042 0.048 5 0.037 0.031 6 0.052 0.064 Mean 0.04 0.041 SD 0.012 0.017 p Value p>0.1 Reference Range 0.002-0.08 Saliva (PPM) SS# Pre Post--2 Months 1 0.029 0.019 2 0.021 0.023 3 0.022 0.072 4 0.022 0.025 5 0.031 0.241 6 0.023 0.029 Mean 0.025 0.068 SD 0.004 0.087 p Value p>0.1 Reference Range Not available
VI - Discussion
One of us (GEA) has previously reported on the physiological, clinical and toxicological aspect of colloidal gold used in amounts of 3 to 30mg/day for several years in normal subjects and for up to one year in ten patients with Rheumatoid Arthritis (RA) (1), (6). There was no evidence of toxicity in the normal subjects and in RA patients studied. Colloidal gold with particle sizes below 10nm had a normalizing effect on body weight, blood platelets, white blood cells and hemoglobin levels (1), (6).
In 5 subjects evaluated with a batteries of tests to assess IQ before and 1 month post colloidal gold at 30mg/day, the mean IQ score increased 20% following colloidal gold (7). In a double blind logitudinal study of 14 elderly subjects aged 65-86 years, there was no significant effect of placebo on the parameters studied in the 6 subjects which served as control. In the 8 subjects on colloidal gold at 20mg/day, there were significant improvements after 4 and 8 weeks on gold of the following: Pain, fatigue, brain fog, physical wellbeing, cognitive wellbeing, overall wellbeing, short term memory, equilibrium and coordination (1).
Because gold is very expensive, the precious metal silver was tested in the colloidal form to assess similarities of beneficial effects between colloidal gold and colloidal silver. Gold, silver and copper belong to the transition group I of the periodic table. Therefore they would be expected to share certain physiological properties. Pilot studies revealed that several milligrams not micrograms amounts of colloidal silver were required to elicit consistent cognitive and physical effects, just like the observations made with pilot studies of colloidal gold more than 10 years previously (1), (6), (7).
The present study involved the evaluation of potential toxicity and beneficial effects of colloidal metallic silver, well characterized in term of particle size and with very low levels of ionic silver; (a ratio of total silver/ionic silver greater than 5000) (3). The daily amounts of 75mg were based on previous experience as mentioned previously. The benefit/risk ratio of colloidal silver depends on the beneficial effects compared with unwanted side effects. The most common side effect reported in humans with the use of ionic silver and mild silver-protein complexes is argyria. Between 1802 and 1951, around a 150 year period, 365 cases of argyria were reported (8). The dosage required to induce argyrosis ranged from 1 to 30 gm by ingestion; and 1 to 8gm by inhalation. No argyria has ever been reported with the use of colloidal metallic silver properly characterized.
All previous studies reporting toxicity of silver were performed with ionic silver. In the management of burn wound sepsis, the ionic silver sulfadiazine cream has been used extensively and successfully but not without side effects (9): argyria, ocular injury, leucopenia; toxicity in renal, hepatic and neurologic tissues. Colloidal silver impregnated gauze is currently been tested as a replacement of the sulfadiazine derivatives (10).
A review of the literature on ionic silver toxicity by Ganther yielded the following information (11). In farm animals and poultry, fed a vitamin E deficient diet, ionic silver, added to drinking water at 0.15% elicited signs of toxicity and muscular dystrophy. A combined deficiency of vitamin E and selenium with added ionic silver to the diet resulted in liver necrosis. The liver necrosis could be prevented with selenium supplementation. Vitamin E at 200 IU/Kg of diet and selenium I PPM prevented silver toxicity. Copper deficiency was induced by ionic silver added to the diet of chicks.
Nordberg and Gerhardsson (12) quoted a 1974 publication, describing the toxic effects in turkey poults of ionic silver acetate at 900mg/kg diet. The addition of selenium, copper and vitamin E to the diet prevented silver toxicity. In susceptible mice, ionic silver induced antinucleolar autoantibody production (5). In a review on the toxicological aspects of topical silver pharmaceuticals for human use, by Hollinger (13), all the ionic silver preparations displayed toxicity. The only metallic colloidal silver tested was without toxic effect although possessing bacteriostatic property.
A recent 2008 publication in the Canadian Journal of Cardiology reported supposedly the first case of cardiomyopathy in humans caused by colloidal gold and silver. S.L Archer from the University of Chicago (14) gave a list of references documenting several side effects of gold-containing drugs, but all these products contains ionic gold, not colloidal gold. He also quoted a 1976 publication by Van Vleet (15) who used very large doses of ionic silver, not colloidal silver, to induce selenium and vitamin E deficiency. A concentration of 0.2 % silver acetate in the ration of pigs did not induce toxicity but 0.5 % of silver acetate resulted in cardiac, hepatic and muscular lesions characteristic of selenium/vitamin E deficiency. These amounts of ionic silver would be the equivalent to 5 to 10 gm of ionic silver/day in an adult human subject, which is one hundred thousand times higher than the amount of silver Archer's patient was consuming.
Daily intake of around 0.1 mg of silver was consumed by Archer's patient on and off for 7 years and colloidal gold 0.1mg/day for 3 months. She developed cardiomyopathy one month after starting the gold product. Serum gold level was below the sensitivity of the assay. Serum silver levels were 1.6 micrograms/L. Our previous experience with colloidal gold, not ionic gold, at 30mg/day for one year in 10 patients with rheumatoid arthritis1'6 revealed that colloidal gold was safe at those amounts and associated with several beneficial effects. Even if the preparations consumed by Archer's patient were in the ionic form, it is unlikely that such small amounts would cause the cardiomyopathy.
Ionic silver can undergo univalent redox reactions with per-oxidativc damage to nearby tissues (14). Antioxidants such as vitamins C and E have a protective effect. Ionic silver can bind to selenium, causing a relative selenium deficiency. This resulted in decreased synthesis of glutathione peroxydase (GP) in kidney, liver and red blood cells (11). This effect of ionic silver on GP is reversible with supplementation with vitamin E and selenium.
Because of the possibility that colloidal metallic silver can undergo in vivo oxidation in a prooxidative environment to yield ionic silver, it was necessary to evaluate the presence of the deleterious effects of ionic silver in the subjects studied with colloidal silver. Anti nucleolar antibody production observed with ionic silver in susceptible mice (5) was not detected in the 6 subjects both before and 2 months post colloidal silver. Mean red blood cell selenium levels increased near significantly (p=0.05) following colloidal silver with mean[+ or -]SD values of 0.208[+ or -]0.02 PPM pre--and 0.225[+ or -]0.03 PPM post 2 months on colloidal silver (Table XII). RBC silver level before intervention was undetectable in 4 subjects (<0.1PPB) and 0.12PPB and 0.2PPB in the other 2 subjects. RBC silver increased significantly from pre-silver mean value of less than 0.12 ug/L to a mean value of 162 ug/L following 2 months of supplementation with silver. It is likely that the increased RBC selenium is due partly to its binding with RBC silver. Since RBC glutathione pcroxydase (GP) levels did not decrease following silver, the binding of silver to selenium was not significant enough to affect the synthesis of GP.
Urine lipid peroxide levels are a good index of the oxidative burden (16). The oxidative burden caused by colloidal silver at 75mg/day was not significant enough to affect the levels of urine lipid peroxides and RBC glutathione peroxydase levels. The levels of urine lipid peroxides were within the reference range for all the subjects prior to and following 2 months on colloidal silver. Red cell selenium and glutathione peroxides levels were within the normal range for all the subjects' pre and post colloidal silver. As a group, there were no significant effects of colloidal silver on urine lipid peroxydes, and red cell selenium and glutathione peroxydase levels.
Mean RDW value decreased near significantly (p=0.07) post-colloidal silver with mean[+ or -]SD in percentage variation of 14.7%[+ or -]1.12 before and 14.32%[+ or -]1.06 post colloidal silver, the upper normal limit being 15% (Table VIII). In subject #3 and #6, the RDW (%) was elevated preintervention, being 16.3% and 15.4%. After 2 months on colloidal silver, these values decreased in both subjects from 16.3% to 15.7% and 15.4 to 14.9%. Iron deficiency anemia is associated with increased RDW, but with normal or decreased MCV (17). In all 6 subjects, the MCV values are within the normal range, but RDW values were in the upper normal or above normal. This is evidence of mild iron deficiency without anemia in the 6 subjects. It is of interest that RBC iron levels increased significantly post-silver (Table XI) from mean[+ or -]SD of 747.8[+ or -]57.5 to 809[+ or -]46 (p<0.01). This increased RBC iron with decreased RDW is compatible with a correction of iron deficiency with silver supplementation. Whether this is due to increased absorption of iron or to increased mobilization from iron stores requires further study. If this normalizing effect of colloidal silver on RBC iron and RDW can be confirmed in a larger group of subjects, colloidal silver could be used to improve iron deficiency even in the absence of anemia.
In subject #5 who experienced muscle pain through the study period, there was a marked increased in urine lipid peroxide levels following silver supplementation (Table XIII), reaching levels of 9.2 micromoles/gm creatinine after 2 months on silver. Although this value is below the upper limit of the reference range, that is 10 micromoles/gm creatinine, it increased to almost 3 fold from a baseline level of 3.6. Subject #5 also experienced a rapid increase in serum alkaline phosphatase levels from 136 IU/L pre silver to 170 IU/L after 2 months on colloidal silver, which is above the reference range of 25 to 165 IU/L. In the other 5 subjects, the rise in alkaline phosphatase following colloidal silver was much less (Fig 3), with values ranging from 62 to 102 IU/L pre-silvcr and a range of 72 to 106 IU/L post 2 months on colloidal silver. The upper limit of serum alkaline phosphatases (165IU/L) may be too high and needs further evaluation in well screened normal subjects, not hospital personel. Compared to the other 5 subjects, subject #5 had an elevated pre-intervention alkaline phosphatases level. If this pattern of elevated serum alkaline phosphatases levels associated with myalgia post colloidal silver can be reproduced in a larger population, this biochemical parameter could have prognostic value. For the time being, blood chemistry including serum alkaline phosphatases levels should be included in the laboratory tests performed before administration of colloidal silver and during follow-up. Total alkaline phosphatases levels are an index of the combined isoenzymes from the liver, bone and intestinal tract. In cases of elevated alkaline phosphatases levels, fractionation of the isoenzymes would be desirable in order to locate the sources of the elevated alkaline phosphatases.
[FIGURE 3 OMITTED]
In the symptomatology questionnaire, subject #5 scored 10 for muscle pain pre-silvcr (no symptoms); 4 after 1 month and 3 after 2 months on colloidal silver. However, she did not consider the muscle pain severe enough to discontinue the colloidal silver. Subject #5 is unique in several respects. The levels of silver in her post-silver saliva was 100 times higher than the other 5 subjects (Table XIV). The level of copper in the post-silver saliva was 10 times higher than the other subjects (Table XV). We have no explanation for this difference between subject # 5 and the others.
Copper deficiency was reported in chicks fed a diet containing ionic silver (11). We did observed a significant (p<0.05) decrease in RBC copper levels with pre-intervention RBC copper mean levels[+ or -]SD was 0.7[+ or -] 0.08 PPM and 0.57[+ or -]0.08 PPM post silver. Prior to supplementation with colloidal silver, RBC copper levels were within the reference range in all the subjects and plasma copper was below the reference range in subject #2 (Table XV). Following 2 months of supplementation with colloidal silver at 75mg/day, RBC copper levels were below the reference range in subject #1 and subject #5. Plasma copper was below the reference range in all but one subject #6. Plasma copper decreased by 45% whereas RBC copper dropped only 15%. Since the decrease in copper levels following colloidal silver was much greater in plasma than RBC, the copper transport protein ceruloplasmin levels may have been suppressed by colloidal silver. Urine copper levels would be expected to increase with lower levels of plasma ceruloplasmin. Since urine copper levels did not increase after colloidal silver, the mechanism of action of colloidal silver in the metabolism of copper must await further research.
Just like silver, the main organ for excretion of copper is the liver. Copper is initially secreted into bile bound to amino acids and biles acids. As the copper passes through the ducts, it becomes complexed with bilirubins, and in this form, copper is unavailable for reabsorption (18). Since scrum bilirubins levels decreased significantly following colloidal silver, the most plausible explanation for the loss of copper following colloidal silver is increased liver clearance of bilirubins and increased biliary excretion of bilirubins complexed with copper which then becomes unavailable for reabsorption.
To our knowledge, this is the first report of a negative effect of colloidal silver on copper metabolism in humans. Copper in trace amounts is required by all living organisms to maintain proper cellular functions. Over 95% of copper in plasma is bound to ceruloplasmin, a copper transport protein that plays many important roles: transport of copper; mobilization of iron from storage site into the plasma (copper deficiency induces iron deficiency anemia); catalytic oxidation of plasma reductants; scavenging of superoxydes; modulation of inflammatory responses (19).
Copper is also involved in several enzymes: cytochrome oxidase, which is the terminal oxidase in the respiratory chain; superoxide dismutase which catalyzes the dis-mutation of superoxide; dopamine betahydroxylase, a critical enzyme in the biosynthesis of norepinephrine, a neurotransmitter; thyrosinase, involved in the synthesis of melanin which is essential for skin pigmentation; Lysyl oxidase which is involved in the cross-linking of collagen and elastin, resulting in increased elasticity of blood vessels (20). copper deficiency results in decreased lysyl oxidase and decreased vascular elasticity. Increased vascular rigidity predisposes to aneurism. From the above information, it is obvious that decreased copper levels in RBC and plasma following colloidal silver is of great clinical importance.
The pro-oxidant effect of ionic silver previously described (11) may be due to ionic silver-induced copper deficiency. The protein primarily responsible for serum antioxidant activity is ccruloplasmin which inhibits markedly lipids peroxidation (19). Chronic copper deficiency is associated with decreased plasma ceruloplasmin levels. Copper supplementation could prevent the pro-oxidant effect of ionic silver. We plan to measure plasma ccruloplasmin levels in our next study.
From a review of the published literature on silver toxicity, the recurrent theme is that toxicity is due to ionic silver induced deficiencies of essential nutrients and supplementation of these nutrients in adequate amounts prevented the toxic effects of ionic silver. It is then obvious that essential nutrients should be administrated to subjects on colloidal silver in daily amounts comparable to the amount used in the present study. We are planning to repeat the RBC and serum silver and copper levels in these 6 subjects after one month on nutritional supplementation.
Although ionic silver was used extensively in the past for its antibacterial effect against a broad range of microorganisms (21 and Vide Infra), it is only in the past 100 years that colloidal metallic silver became popular among clinicians and the public at large for its germicidal properties . Recent in vitro studies with ionic silver suggest that its antibacterial effect is due to its interference with the respiratory chain at several sites (22-24) and its destabilizing effects on intermolccular adhesions (25). In the last few years, some studies were published describing the mechanisms of action of colloidal silver against bacteria and virus. The mechanism of action of colloidal silver is totally different than the mode of action of ionic silver, which binds covalently to thiol groups of various enzymes, blocking their action. On the other hand, colloidal silver binds non-covalently to macromolecules, altering their biological properties. The effect of ionic silver is charge dependant whereas the size of colloidal silver particles plays the key role (1).
In 2005, Morones, et al, (26) reported the results of their experiments with the use of colloidal metallic silver. These investigators studied the effect of silver colloids in the range of 1-100 nm on gram-negative bacteria inoculated on agar plates. Only silver nanocolloids less than 10 nm were able to bind to the bacterial membrane and penetrate inside the bacteria. They reported that these silver nanocrystals were spherical in shape. The overall effect of metallic silver nanopartieles was different from the effect of silver ions which acted mainly at the surface of the membrane and triggered a proteetive meehanism in the bacteria. The metallic silver nanocrystals did not trigger this protective meehanism and were able to penetrate inside the bacteria. To summarize their findings, spherical silver nanopartieles in the range of 1-10 nm attach to the surface of the bacterial cell membrane and drastically disturb its proper function, like permeability and respiration. These silver nanopartieles are also able to penetrate inside the bacteria and cause further damage by possibly interacting with sulfur and phosphorus containing compounds. A recent study of the effect of silver nanopartieles on HIV-1 confirms the 10 nm size limit for effectiveness. Elechigucrra, et al, (26) used silver nanopartieles with a mean diameter of 21 nm but with a wide range of sizes. They demonstrated that silver nanopartieles undergo a size-dependent interaction with HIV-1, with nanopartieles exclusively in the range of 1-10 nm binding to the HIV-1 at the exclusion of larger colloidal particles.
Considering the current popularity of colloidal silver, it is surprising that practically nothing is known about its metabolism and about its effect in clinical conditions not related to infectious diseases. We previously reported serial measurements of blood silver concentration over a 24hr period analyzed by neutron activation analysis (NAA) in 5 female volunteers (3). The sensitivity of NAA for silver is 10ug/L. The whole blood silver levels were below 10ug/L before ingestion of colloidal silver. Between 2 and 10 hours post silver ingestion, the whole blood levels ranged from 18-68ug/L. Besides our previous publication (3), no data are available on the blood levels of silver following ingestion of colloidal silver.
Jensen, et al, (28) reported in 1988 the serum levels of silver in 21 subjects who consumed daily 30mg of the ionic silver actate from anti-smoking chewing gum for a period of 12 weeks. The mean and ranges of the serum silver levels were: Before intervention: 2.8ug/L (Range 1.5-3.5); After 12 weeks of intervention: 55ug/L (Range 26-87); and 14 weeks after the end of the study: 3.2 (Range 2.6-4.5). Since silver acetate contains approximately 65% silver, Jensen's subjects consumed 19.5mg ionic silver/day. Our subjects consumed 75mg of colloidal silver/day, 4 times the daily amounts of ionic silver used in Jensen's study. After 2 months of colloidal silver, the mean values for our 6 subjects were: RBC Silver = 162ug/L; Plasma Silver = 161ug/L (Table XIV). These plasma and RBC values are 3 times higher than the mean values of 55ug/L of serum observed in Jensen's subjects who ingested 4 times less silver than our subjects. Plasma values of our subjects are comparable to Jensen's serum values, when the amounts of silver ingested are taken into consideration.
The metabolism of ionic silver has been reported previously (12)(29), (30). Less than 10% of ingested ionic silver salts is absorbed. Approximately 90% of the absorbed silver is eliminated in the feces after biliary secretion. Less than 1% of ingested silver is excreted in urine. When ionic silver is injected intravenously in rats, rabbits, and dogs (30), 80% of the dose was excreted into the feces over a period of 4 days. Less than 0.5% was detected in urine. The similarities and differences between the metabolism of ionic silver and colloidal silver must await further research in this area.
The liver is the major organ of silver elimination and contains the highest concentration of silver, followed by spleen, skin, muscle and brain. Nutritional supplementation besides vitamin E, selenium and copper should include nutrients involved in maintaining normal liver functions. This is our rationale for recommending a complete dietary program when colloidal silver in amounts comparable to those used in the present study are ingested for a long time. The only side effect of ionic silver not known to be associated with nutritional deficiencies is argyria, which is a cosmetic complication without any clinical significance.
The authors wish to express their gratitude to Ralph M. Albrecht and Joe Heins for performing the TEM particle sizing and the ultracentrifugation; to the volunteers who participated faithfully in this project; and Ameen Lalani and Regina Jauregui for typing the manuscript.
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by: Guy E. Abraham, MD and Jorge D. Flechas, MD
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|Author:||Abraham, Guy E.; Flechas, Jorge D.|
|Date:||Sep 1, 2009|
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