Rest in peace nanobacteria, you were
not alive after all
Nanobacteria are Not a Cause of Calcification in Arterial Plaque of Cardiovascular Disease
by Elmer M. Cranton, M.D.
Recent research in 2008 gives evidence that so-called nanobacteria consist of tiny calcium crystals and are not living organisms
It had been erroneously hypothesized that a new life-form named "nanobacterium" was responsible for these findings.
Pathologic calcification of atherosclerotic plaque, dental plaque and kidney
stones has been theorized by Finnish researchers, Kajander and Ciftcioglu
to be caused by a previously unreported bacterial species, tentatively named
Nanobacterium sanguineum. Scientists at NIH and the FDA then
duplicated that research and, although laboratory observations were identical,
they discovered much simpler explanations without need to invoke a living
organism. Government scientists concluded that the calcific
mineralization observed in the laboratory was easily explained by the
nucleating properties of self-propagating but non-living microcrystalline
centers (nidi), which form crystalloid macromolecules of calcium carbonate
Submicroscopic (submicron) apatite crystals can be transferred in a deceptively life-like manner through serial dilutions, using techniques similar to those used for bacterial subcultures, while retaining their crystalline ability to grow into calcific deposits at body pH. Six serial 1:10 dilutions of these crystalline nidi were transferred to fresh culture media and repeatedly found to propagate as tiny coccoid or dome-shaped calcific crystals. Photographs of these non-living structures taken under electron microscopy were identical in appearance to those previously published by the Finnish researchers and hypothetically labeled as “Nanobacteria.”
The 16S rDNA chromosomal sequences attributed by Kajander and Ciftcioglu to Nanobacteria sanguineum were identified as belonging instead to a common environmental microorganism, Phyllobacterium mysinacearum, a known contaminant of culture media and laboratory reagents—not associated with calcification. Although reagents and culture media are sterilized before use, traces of these DNA sequences remain, which are greatly amplified in the laboratory procedure. Those same 16S rDNA sequences were found in control specimens, which had no potential to be a source of a living organism such as the imputed Nanobacteria. No controls were reported by the Finnish researchers, who instead assigned these 16S rDNA sequences to a new bacterial genus.(1)
Examination of calcific biofilms identical to those previously described by Kajander and Ciftcioglu revealed no nucleic acids or proteins of the type that would be present in bacteria. Progressive calcific propagation was not inhibited by sodium azide, an extremely potent cellular and bacterial toxin. Propagation was inhibited by the tetracycline, but tetracycline is a calcium chelator and would be expected to inhibit crystal growth by that mechanism.
High-dose gamma radiation and ultra filtration to the submicron level did prevent calcific propagation, but that is not evidence for living bacteria. Those two procedures alter macromolecular and crystalline properties in solution, removing microcrystalline nidi, blocking further growth of calcific crystals.
Inorganic calcium and phosphate ions, when added to sterile solutions of culture media, also resulted in progressive calcific mineralization. Phospholipids, lipid-protein complexes, and submicroscopic crystals of calcium apatite occur in plasma and were all found to be nucleators of calcific biomineralization.
Cell cultures of fibroblasts grown in the presence of these calcific biofilms showed evidence of cytotoxicity and were observed to take up inorganic microcrystals of calcium apatite. The fact that intracellular calcification is poorly tolerated is well known and that observation cannot be interpreted as evidence for the presence of Nanobacteria.
Antibodies can be produced to react with surface structures of nonliving
crystalline macromolecules using monoclonal techniques. Chelating agents such as
EDTA, citrate, and tetracycline (an antibiotic that also has calcium binding
properties) can alter the surface properties of nonliving calcific and
crystalline nucleators—both halting calcific propagation and blocking
immunologic reactivity to previously reactive antibodies. Diagnostic testing
based on immunologic properties could thus become non-reactive following
exposure to calcium chelators. That change therefore does not indicate that a
bacterial infection has been eliminated, as claimed by one laboratory.
There has been a recent flurry of marketing activity attempting to sell EDTA suppositories to unwary medical practitioners and patients―even by health food stores to the general public. Proponents of those products base their sales pitch on the unproven theory that Nanobacteria cause arterial plaque and that EDTA suppositories will remove the alleged "calcific shields," supposedly produced by Nanobacteria as protection. Proponents of EDTA suppositories further claim that a powder must be taken by mouth to delay the renal excretion of EDTA, resulting in higher blood levels. None of those claims are credible for the following reasons:
1. Nanobacteria (if they even exist) have not been shown to
have any relationship to pathologic calcification in the body as found in
2. EDTA is 100% filtered by renal glomeruli. The filtration rate of EDTA equals the glomerular filtration rate (GFR). EDTA is not subject to tubular secretion or reabsorption. The only way to delay excretion of EDTA would be to poison the kidneys sufficiently to reduce creatinine clearance (GFR)—not a wise thing to do.
3. No non-metabolized drug such as EDTA is better absorbed rectally than by mouth. Suppositories can be used in cases of persistent vomiting, in uncooperative patients who refuse to take oral medicines, and in pediatric patients who will not swallow medicines by mouth. Passive absorptive properties of the colon and rectum are similar to the upper G/I tract. It is well documented that orally administered EDTA is not absorbed in significant amounts—approximately five percent. Oral EDTA therefore ends up in the rectum, right where a suppository would be inserted. If rectal or colon uptake of EDTA were greater than in the upper G/I tract, then total absorption by mouth would be greater than 5%. It has been scientifically demonstrated that both rectal and oral absorption of EDTA is at most seven percent.
4. The "secret formula" powder recommended to be administered by mouth to enhance rectal EDTA also contains additional oral EDTA. It may therefore add somewhat to total absorption, but not by delaying renal excretion, as asserted by the marketers of this product. Blood levels of EDTA that were reported using this combination of oral and rectal EDTA are less than one sixth that achieved by intravenous infusion. It is true that 12 hours later the small amount of EDTA continuing to be absorbed from the gut will produce blood levels at that time which exceed what would remain from a rapidly-excreted intravenous infusion. It is untrue, however, to state that the EDTA suppositories combined with oral EDTA result in a higher blood level of EDTA using this type of deceptive data.
5. Eighty-five percent of patients have consistently responded well to intravenous EDTA, administered according to the protocol developed and widely accepted over almost 50 years of continuous use. Electron beam CT (EBCT) scores of coronary artery calcium usually remain unchanged while patients improve dramatically. Many published studies have presented objective evidence of increased blood flow and improvement in symptoms and function, with no change in EBCT calcium scores. It therefore makes little sense to assume that change in calcification of artery walls is an important indicator of clinical improvement. Arterial plaque is a proliferative and inflammatory disease, involving soft tissue cell replication during most of its course. Any experienced chelation clinic has accumulated large numbers of case histories showing dramatic improvement following the well-established protocol of intravenous EDTA. Do proponents of EDTA suppositories and oral EDTA believe that intravenous treatment is not effective, despite these proven benefits reported in dozens of published studies? If another party proposes an innovative method to produce similar results, they should publish data from a series of consecutively treated patients with tabulated, objective and statistically significant evidence to document both effectiveness and safety of their position. It need not be a blinded, controlled study―a mere 10 consecutive patients with objective before and after measurements showing benefit would be quite convincing.
6. While EBCT calcium scores do correlate with presence of plaque, they do not correlate well with degree of occlusion. Exaggerating to make a point, wearing a skirt correlates very highly with the incidence of breast cancer. Using this same type of reasoning, it might seem that if women stopped wearing skirts and wore only slacks, they would not get breast cancer. In other words, correlation does not mean cause and effect. Other more important variables are clearly involved. It is known that prolonged administration of EDTA, 50 to 100 infusions, can reduce pathologic calcification and even dissolve kidney stones. But EDTA has other profound effects in the body, not just on undesirable calcium, and with far fewer treatments. EDTA also binds a spectrum of essential nutritional trace elements, often with a 10 fold greater effect than on calcium. There is no reason to assume that pharmacologic blood levels of EDTA sustained continuously for prolonged periods of time are safe (as implied by proponents of both oral and rectal EDTA). In fact, it seems logical to believe that eventual disruptions will occur to vital metalloenzyme systems. If EDTA is continually present in the digestive tract it will bind to nutrients, interfering with nutritional uptake of essential trace metals, leading to deficiencies.
7. The electron beam can only measure discrete slices, and does not cut through exactly the same location on repeat measurement. Variability between examinations of EBCT calcium scores ranges as high as plus or minus 38%, depending on the test site and equipment calibration.(3)
8. The Nanobacteria hypothesis ignores the well-established relationship of plaque to homocysteine levels.
An infective organism may play some role in the course of atherosclerosis―Chlamydia,
for example. Whether it is a late-stage, secondary factor or a causative agent
has yet to be determined. Recent studies have failed to show benefit from a
variety of antibiotics.
In the absence of new evidence to the contrary, Nanobacterium sanguineum seems an unproven hypotheses, that is being used in a clever and deceptive marketing scheme for EDTA suppositories.
1. Kajander EO, Ciftcioglu N. Nanobacteria: an alternative mechanism for pathogenic intra- and extracellular calcification and stone formation. Proc Natl Acad Sci USA. 1998 Jul 7;95(14):8274-9.
2. Cisar JO, Xu DQ, Thompson J, Swaim W, Hu L, Kopecko DJ. An alternative interpretation of nanobacteria-induced biomineralization. Proc Natl Acad Sci USA. 2000 Oct 10;97(21):11511-5.
3. Nieman K, van Geuns J, Wielopolski P, et al. Noninvasive coronary imaging in the new millennium: A comparison of computed tomography and magnetic resonance techniques. Rev Cardiovasc Med 2002 3(2):77-84.)
Copyright © 2007 Elmer M Cranton, M.D.
Copyright © 2012 Elmer M. Cranton, M.D., all rights reserved