Boron and Granulomas (TB, Rheumatoid arthritis, Crohn's, Rheumatic Fever etc.) - 2015

Anti-Inflammatory Effects of Boron Alone or as Adjuvant with Dexamethasone in Animal Models of Chronic and Granulomatous Inflammation
Hanaw N. Ameen, 2015 Thesis, University of Sulaimani ,Kurdistan Iraq, Master of Science in Pharmacology
 the entire thesis PDF is in VitaminDWiki

Published paper with identical tile
 the 7 page resulting paper is also in VitaminDWiki
Abstact
Background: Side effects of anti-inflammatory agents are a major problem during clinical use. The development of a newer, effective, and safe anti-inflammatory agent should be considered. Boron-containing compounds are found effective as anti-inflammatory agents with relatively low side effects. We aimed to evaluate the anti-inflammatory activity of boron in animal models of chronic and granulomatous inflammation.

Methods: Sixty-six Wistar rats were allocated into five groups; 1st (6 rats) treated with vehicle only without induction as a negative control; 2nd (12 rats) allocated into two subgroups, treated with vehicle only, with induction of chronic and granulomatous inflammation, as appositive control. 3rd group (24 rats) allocated into four subgroups, treated with different doses of boron (3 and 6 mg/kg) in both models. Fourth group (12 rats) treated with dexamethasone (1 mg/kg) in the same models. 5th group (12 rats used) treated with boron (3 mg/kg) with dexamethasone (1 mg/kg) in the same models.

Results: Boron, in a dose-dependent pattern significantly decreases inflammation in rat models of chronic and granulomatous inflammation.
Combination of boron with dexamethasone significantly suppresses inflammation in both models, which is significantly higher than all of the effects produced by other approaches of treatment.

Conclusion: Boron, in a dose-dependent pattern, effectively suppresses formaldehyde-induced chronic inflammation and cotton pellet-induced granuloma in rats when used alone or as an adjuvant with dexamethasone. It may be considered as a potential treatment for chronic inflammatory conditions.

Both the thesis and the paper were published in 2015

Granuloma in Wikipedia

• "Granulomas form when the immune system attempts to wall off substances it perceives as foreign but is unable to eliminate."
• Diseases include: Tuberculosis, Leprosy, Schistosomiasis, Histoplasmosis, Cryptococcosis, Cat-scratch disease
  Rheumatic Fever, Sarcoidosis, Crohn's disease, Listeria monocytogenes, Pneumocystis pneumonia
  Aspiration pneumonia, Rheumatoid arthritis, Granuloma annulare, Foreign-body granuloma

Dexamethasone in Wikipedia

• " , , steroid medication.It is used in the treatment of many conditions, including rheumatic problems, a number of skin diseases, severe allergies, asthma, chronic obstructive lung disease, croup, brain swelling, and along with antibiotics in tuberculosis"
• "It may be taken by mouth, as an injection into a muscle, or intravenously"
• "Dexamethasone is not expensive. In the United States a month of medication typically costs less than 25 USD. In India a course of treatment for preterm labor is about 0.5 USD."

   Bolding by VitaminDWiki

Boron, the Element

Boron, ubiquitous in the earth’s crust, can be found in most soil types as well as in fresh and salt water. While most of the earth's soils have <10 ppm Boron, the range is from 2-100 ppm with the average soil Boron concentration reported to be 10-20 ppm. While large areas of the world can be Boron deficient, high concentrations can also be found, for example, in parts of the western United States, throughout China, Brazil and Russia. The world’s richest deposits of Boron are located in a geographic region that stretches from the Mediterranean countries inland to Kazakhstan. Seawater contains an average of 4.6 ppm Boron, but ranges from 0.5-9.6 ppm. Freshwaters normally range from <0.01-1.5 ppm, with higher concentrations in regions with high concentrations of Boron in soil [27]. Most essential elements that make their way into the human food and water supply are directly derived only from soil minerals. While most environmental sources of Boron are geogenic in nature, some trace elements such as Boron, iodine, and selenium are supplied in significant amounts to soils by atmospheric transport from the marine environment.

Deficiency problems associated with these elements are therefore generally less common in coastal areas than farther inland. It has been known for some time that Boron is an essential micronutrient for higher plants yet the mechanism through which Boron functioned in plants was, until recently, unknown [28,29]. While Boron accumulates in aquatic and terrestrial plants it does not magnify through the food-chain. On the other hand, its deficiencies in plants are often observed. Boron is also a constituent in all phyla of living organisms and its role is in most obscure [30]. For some microorganisms, algae, and higher plants, Boron is essential. Although the quantities required are low they are also highly variable and species specific. In other species, including humans, knowing how much Boron is needed and what Boron does is still being determined.

Boron has long been recognized as an essential trace element for plants, but has only recently been considered to be possibly essential for humans. Boron appears to participate in hydroxylation reactions, which play a role in the synthesis of steroid hormones and vitamin D. In Australia, where much of the food is grown on soil deficient in this mineral, Boron supplements were popular as a treatment for osteoarthritis (OA), and were reportedly selling at a rate of 10,000 bottles per month before the Australian government removed the product from the market [30]. In a double-blind study, 20 Australians with OA were randomly assigned to receive Boron (6 mg per day as sodium tetraborate decahydrate) or a placebo for eight weeks [32]. Of those receiving Boron, 50 percent improved, compared with 10 percent of those given placebo. Because of the small sample size, this difference was not statistically significant. When the five subjects (25%) who dropped out of the study (mostly because of clinical deterioration) were excluded from the analysis, 71% of those in the Boron group improved, compared with 12.5 percent of those in the placebo group (P< 0.05). No side effects were seen and there were no significant changes in common laboratory parameters. These results suggest Boron supplementation may be helpful for individuals with OA whose diets are likely to be low in Boron.

Further research is needed to confirm this preliminary study and to determine whether individuals with a higher dietary intake of Boron can benefit from supplementation. The average American diet provides approximately 1-2 mg of Boron per day, primarily from fruits, vegetables, and nuts; however, according to German research, intake can vary from 0.3 to 41 mg per day. While the capacity of Boron to increase estrogen levels [33] might raise concerns about possible cancer risks with Boron supplementation, there is no evidence that populations with a high intake of Boron (such as the French) have an increased incidence of hormone-related cancers.

Pharmacokinetics of Boron

Boron is easily absorbed across the gastrointestinal epithelia in humans and animals [34], and across mucous membranes, such as the mouth, eyes, vagina, and anus. In 1998, Hunt reported that humans and animals absorb nearly 100% of supplemental inorganic Boron. Some organic forms of supplemental Boron may be inaccessible to animals because plants can only absorb organic forms of Boron in soils after mineralization [35]. Boron is primarily excreted in the urine, with about 2% lost in the feces, and lesser amounts lost in bile, sweat, and breath [36,37]. Tissue Boron concentrations are generally kept steady by a homeostatic mechanism, primarily through renal excretion, and higher Boron intakes do not significantly increase plasma levels [38]. A 167-day metabolic study of 11 postmenopausal women showed a rapid increase in urinary Boron when Boron intake increased from (0.36 mg/day) to (3.22 mg/day) [39]. Naghii and Samman [40] studied the effect of Boron supplementation on urinary excretion in healthy male subjects. When 18 healthy males remained on a habitual diet, urinary Boron excretion measured on two separate occasions ranged from 0.3 to 3.53 mg/day.

The difference in Boron values between the two 24-h urinary collections was not statistically significant, but slight variations within and between some subjects suggested differences in their daily Boron consumption. In a second study, when subjects were administered 10 mg/day of supplementary Boron for 4 weeks, urinary Boron increased from an average of 1.64±0.3 (at baseline) to 10.16±0.92 mg/day. This increase in urinary excretion, which occurred in every individual, was significant and represented 84% of the supplemented dose. These findings provide evidence that urinary Boron reflects Boron intake.

Role of Boron Intake on Health of the Population

The suggestion that Boron may be a factor in maintaining health is reasonable because there is evidence that many people consume less Boron than the necessary to promote bone and brain health. In human depletion-repletion experiments, subjects responded to a Boron supplement after consuming a diet supplying only 0.2-0.4 mg Boron/day for 63 days [41] suggesting that this intake of Boron is inadequate. Thus, a dietary Boron intake higher than 0.4 mg/day may be beneficial to bone and brain health. Extrapolation of data from animal experiments suggests that 1 mg Boron/day would provide optimal nutritional benefits for this element. The WHO suggested that an acceptable safe range of population mean intakes of Boron for adults could well be 1.13 mg/day [42] relying on both animal and human data. Based on published values for Boron in foods, it has been estimated that the median intake of Boron in the United States is 0.86 mg/day [43]. The 1994-1996 Continuing Survey of Food Intakes by Individuals indicated that Boron intakes ranged from a low of about (0.35 mg/day) to a high of about (3.25 mg/day) for adults [44]. The median intakes for various age groups of adults ranged from (0.87 to 1.13 mg/day). The reported median intakes of 0.86 and 0.87 mg Boron/day suggest a significant number of people would benefit from increased Boron intakes. This suggestion is supported by a study of premenopausal women in eastern North Dakota [45]. Based on urinary excretion of Boron (a good indicator of Boron intake), two women apparently consumed an average of less than 0.5 mg Boron/day, and 14 women consumed between 0.5 and 1.0 mg Boron/day. There are only a few reports associating Boron intake or status with diseases other than some types of cancer described above. Low concentrations of Boron in hair [46] and low environmental Boron [47] have been associated with Kashin-Beck disease (Osteochondropathic) in China. Low Boron status has been associated with rheumatoid arthritis (RA) [48]. Based on the suggestion that a significant number of people may have a low Boron status, more epidemiological studies determining whether a low Boron intake is associated with some disorders of bone and brain seems prudent.
There are two reports describing no or limited responses by postmenopausal women to Boron deprivation, which may have resulted in negative impressions about the nutritional importance of Boron. Several aspects of the experimental designs of these studies, however, may have contributed to the lack of marked findings. In one experiment, the subjects were only equilibrated on the experimental diet for two days before starting the low dietary Boron regimen that lasted only 21 days [49]. The data (i.e. increasing urinary calcium) presented from only six subjects suggest that they were still adjusting from their self-selected diets to the experimental diet, and thus to changes in other nutrient intakes when they began receiving Boron supplementation of 21 days duration. Additionally, 21 days is an extremely short deprivation period for an adult organism when the diet is not severely deficient and a small number of subjects limits statistical power.

In successful Boron deprivation experiments, 14 subjects were equilibrated to the experimental diet for 14 days, and the first 21 days of Boron deprivation were not included in the analysis because only minimal responses occurred during this time; the most marked effects were seen after 42 days of Boron deprivation [41,50]. Thus, short Boron deprivation periods of only 42 days in a Latin-square experimental design most likely contributed to finding a limited number of responses to Boron deprivation compared to other human studies [51]. In addition, varying dietary magnesium (deficient and adequate) may have obscured or blunted the effects of varying dietary Boron. These design concerns suggest that these two human studies are ill suited for assessing the nutritional relevance of Boron.

Many epidemiological and controlled animal and human experiments have provided evidence for the use of Boron as a safe and effective treatment for some forms of osteoarthritis (OA) [52]. By examining the relationship between Boron administration and OA prevalence around the world, researchers have discovered that in the areas where Boron intake is 1 mg or less per day, the estimated incidence of arthritis is between 20% and 70%. In contrast, in areas where Boron intake is usually 3-10 mg per day, the arthritis percentage is lower, ranging from zero to only 10%. This remarkable finding is a compelling evidence of the fact that abundant intake of dietary Boron can confer strong protection against the development of OA [53,54]. An analytical study showed that Boron concentration is lower in femur heads, bones, and synovial fluid of OA patients as compared with patients without OA. Moreover, surgeons have observed that the bones of patients that had used Boron supplementation were harder to cut than those patients who had not used these supplements [55].

The most convincing evidence for Boron usage in the case of OA patients comes from a double-blind placebo Boron supplementation trial conducted in Australia [56,32] reporting that Boron supplementation may improve symptoms for people with OA and rheumatoid arthritis [32]. Experimental studies on arthritic rats have led to an emerging hypothesis suggesting that Boron reduces the risk of inflammatory disease by downregulating enzymes of the inflammatory response and has a beneficial immunomodulatory effect in the arthritic rats [57-59].

C-reactive protein (CRP), one of the most useful markers of systemic inflammation, has recently been identified as a marker of OA with clinical significance. CRP levels are moderately high for patients with OA as compared with the normal controls [60,61]. Of great clinical significance are CRP levels, with reference values below 0.5 mg/dL in OA patients [62,63]. Increased levels have been associated with the disease evolution as well as with the clinical aggravation, as an unspecific response to inflammations and infections [64-66].

Calcium fructoborate (CF) is used as a recent non-pharmaceutical therapy for osteoarthritis treatment. CF is a complex of calcium, fructose, and Boron and is naturally found in fresh and dried fruits, vegetables, herbs, and wine. This form of Boron is not only safe but also bioavailable compared with other commercial forms of Boron. An open label pilot study, authored by Miljkovic and colleagues from the Orthopedic Clinic of the University of Novi Sad, Yugoslavia, was conducted. The purpose of the study was to investigate the effects of CF on OA symptoms. The study included 20 patients with mild, medium, or severe forms of OA.

Two criteria for assessment were used: the Western Ontario McMaster University Osteoarthritis Index and Newnham criteria. After the administration of CF, the results were quite impressive: the pain was strongly diminished, the joint rigidity disappeared, and mobility and flexibility were improved [67,68]. Previous investigations have been summarized in two other reviews [67,68] that have revealed an antiinflammatory property of CF on cellular cultures. In addition, it has been hypothesized that CF might have dual roles as both an anti-inflammatory and anti-oxidant agent, with modifying effect on lipid metabolism [67,68].

The study investigates whether CF can relieve OA symptoms in selected subjects. Scientists have hypothesized that CF may have a role in diminishing inflammation-related pain, joint stiffness and other discomforts associated with OA [69-71]. Because OA discomfort is often invariably related to joint inflammation, this study approaches the CF effect on inflammatory blood markers levels such as CRP, fibrinogen, and on erythrocyte sedimentation rate (ESR) and on lipid metabolism markers because it has been suggested that Boron is involved in both mechanisms [67]. When analyzing inflammatory markers, the 2-week time interval for the CF dietary supplementation was long, enough to confirm previous results obtained in vitro. Because the general characteristic of the placebo effect has a slightly delayed onset, and a relatively short duration (from 2 to 6 weeks as cited in the literature) [72], a time interval of 2 weeks was chosen for this trial to more accurately observe the short-term efficacy of CF. This pilot study is only a bridge for a future, more complex research study regarding the effects of CF on OA symptoms.

Suggested Mechanisms for the Biological Effects of Boron

The diverse responses reported for animals and humans-deprived of Boron have made it difficult to identify a primary mechanism for its possibly beneficial activity. The wide range of responses is likely secondary to Boron influencing a cell signaling system and/or the formation and/or activity of entity that is involved in many biochemical processes. A plausible mechanism of action may be indicated by the biochemistry of Boron. Boric acid forms ester complexes with hydroxyl groups of organic compounds, which preferably occurs when the hydroxyl groups are adjacent and in a cw-orientation. This property results in the formation of complexes with several biologically important sugars. These sugars include ribose, which is a component of adenosine. Recent findings suggest that the diverse beneficial effects of Boron occur through affecting the presence or action of biomolecules containing adenosine or formed from adenosine precursors. These biomolecules include S-adenosylmethionine and diadenosine phosphates that have higher affinities for Boron than any other recognized Boron ligands in animal tissues [73].

Diadenosine phosphates are present in all animal cells and function as signal nucleotides involved with neuronal response. S- adenosylmethionine is one of the most frequently used enzyme substrates in the body [74]. About 95% of S-adenosylmethionine is used in methylation reactions, which influence the activity of DNA, RNA, proteins, phospholipids, hormones, and transmitters. The methylation reactions result in the formation of S-adenosylhomocysteine, which can be hydrolyzed into homocysteine. Support for the hypothesis that Boron bioactivity is through an effect on S-adenosylmethionine formation and/or utilization are the findings that plasma homocysteine increased and liver S-adenosylmethionine decreased in rats fed (0.05-0.15 mg/kg) Boron compared to rats supplemented with (3 mg/kg) diet [75]. High circulating homocysteine and depleted S-adenosylmethionine have been implicated in many of the disorders that can be affected by nutritional intakes of Boron, including arthritis, osteoporosis, cancer, diabetes, and impaired brain function.

Further support for the hypothesis is that the bacterial quorum sensing signal molecule, autoinducer-2, is a furanosyl borate ester synthesized from S-adenosylmethionine [76]. Quorum sensing is the cell-to-cell communication between bacteria accomplished through the exchange of extracellular signaling molecules (auto-inducers). Moreover, Boron strongly binds the oxidized form of nicotinamide adenine dinucleotide (NAD+) [73], and thus might influence reactions in which it is involved. One role of extracellular NAD+ is to bind to the plasma membrane receptor CD38, an adenosine diphosphate ribosyl cyclase that converts NAD+ to cyclic ADP ribose. Cyclic ADP ribose is released intracellularly and binds to the ryandodine receptor, which induces the release of calcium ions from the endoplasmic reticulum. Cell culture studies show that Boron binds to and is a reversible inhibitor of cyclic ADP ribose [77,78]. Concentrations of Boron found in blood are found to decrease Ca2+ release from ryandodine receptor-sensitive stores [78]. Thus, it has been hypothesized that Boron is bioactive through binding NAD+ and/or 2+cyclic ADP ribose and inhibiting the release of Ca , which is a signal ion for many processes affected by Boron, including insulin release, bone formation, immune response, and brain function.

Studies with plants have resulted in another suggested plausible mechanism of action for Boron bioactivity. Boron might be bioactive through forming diester borate complexes with phosphoinositides, glycoproteins, and glycolipids in cellular membranes. Diester borate polyl complexes might act as calcium chelators and/or redox modifiers [79] that affect membrane integrity and function [80]. This modifying effect could alter the transduction of regulatory or signaling ions across membranes. Determination of such an effect in animals and humans has yet to be determined. However, the finding that the borate transporter NaBC1, which apparently is essential for Boron homeostasis in animal cells, conducts Na+ and OH- across cell membranes in the absence of Boron [81], supports the suggestion that Boron deprivation might affect the transduction of regulatory and signaling ions across cell membranes.

Boron and the Inflammatory or Immune Response

Several laboratories have found that Boron status affects the response to injury or infection. Among the findings is that of Boron status affecting the response to the injected antigens. When injected with an antigen (M butyricum in mineral oil) to induce arthritis, Boron-supplemented (2.0 mg/kg diet) rats had less swelling of the paws and lower circulating concentrations of natural killer cells and CD8a+/CD4- cells than did Boron-deficient (0.1 mg/kg diet) rats [57]. Another study found that Boron supplementation (20 mg/kg diet) of a Boron-low (0.2 mg/kg) diet significantly delayed the onset of adjuvant-induced (M. tuberculosis) arthritis in rats [57]. Pigs fed a Boron-low (1-2 mg/kg) diet for 95 days exhibited a significantly higher skinfold thickness response to an intradermal injection of phytohemagglutinin than pigs supplemented with Boron (5 mg/kg diet) [82]. Physiological amounts of Boron (3 mg/kg) supplemented to a Boron-low diet (0.2 mg/kg) more than doubled the serum total antibody concentrations to injected antigen (human typhoid vaccine) in rats [83].

The suggestion that Boron may have a regulatory role in the inflammatory or immune response is supported by a study of mice infected with the nematode H. bakeri [84]. Boron deprivation down- regulated 30 of 31 cytokines or chemokines associated with the inflammatory response six days post-primary-infection. An opposite pattern was found, especially 21 days post-challenge; mice consuming low and marginal Boron-deficient diets had >100% increases in 23 of 31 cytokines determined. This finding is consistent with lower serum TNF-a and INF-y after lipopolysaccharide injection in pigs fed a marginal Boron-deficient diet than in pigs supplemented with a 5 mg Boron/kg diet [85]. Boron also affects changes in immune cell populations induced by other dietary factors, which include dietary fatty acids. Supplementation of young healthy men with 6 g/day of the n-3 polyunsaturated fatty acid docosahexaenoic acid for 12 weeks decreased the number of white blood cells, mainly because of a decreased granulocyte number; the decreased granulocyte number resulted in an increased percentage of lymphocytes in the white blood cells [86]. In contrast, 1.5 g of the n-6 polyunsaturated fatty acid increased granulocyte numbers [87]. Compared with safflower oil (mostly n-6 polyunsaturated fatty acids), fish oil (high in n-3 polyunsaturated fatty acids) increased white blood cell numbers, with most of the increase in the lymphocyte fraction, in Boron-adequate (3 mg/kg diet) rats but not in Boron-deprived (0.1 mg/kg diet) rats [88]. Fish oil instead of safflower oil increased monocyte and basophil numbers in Boron-deprived but not in Boron-adequate rats. Similarly, canola oil (high in n-3 fatty acids) increased the percentages of white blood cells that were basophils and monocytes in Boron-deprived rats, but not in Boron-adequate rats [89].
An effect on the inflammatory response might be the reason that Boron was found beneficial in a study of 20 patients with radiographically confirmed osteoarthritis consuming daily either a 6 mg Boron supplement or a placebo for 8 weeks in a double-blind trial [32]. The Boron-supplemented arthritic individuals self-reported substantial improvement in subjective measures of joint swelling, restricted movement, and fewer analgesics for pain relief. Affecting the immune response might be the reason that Boron intake has been associated with some cancers, for example breast cancer.

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All of the references are in  the thesis PDF on VitaminDWiki