Effect of NC-518® Calcium on Bone Mineral Density in Men and Women with Osteoporosis and Osteopenia

Background Osteoporosis is characterized by low bone mineral density (BMD) and leads to brittle bones and a heightened risk of fracture. According to the National Osteoporosis Foundation (NOF), osteoporosis affects 10 million Americans. 34 million more are estimated to be at risk of developing the disease due to low bone mass. It is a serious medical condition that has accounted for 2 million fractures in the United States. By 2025, osteoporosis related fractures are expected to rise to 3 million. One in two women and one in four men over the age of 50 will have osteoporosis related fractures in their remaining 1 lifetimes. One of the major contributing factors in the development of osteoporosis is the inadequate dietary intake of calcium and vitamin D. Calcium is essential for the body to function. It plays a vital role in the homeostasis of bone and teeth and ensures that blood clotting systems, nerves, and the heart function properly. Although food is the recommended source of calcium, nearly all adults and children do not 2 consume enough of the daily recommended dose of calcium. Indeed, calcium supplementation is necessary for most adults to even reach the daily recommended dose. Calcium supplements in the market have caused much dissatisfaction due to their negative side effects such as gas, constipation, and upset stomach. The absorption and bioavailability of these products have also been questioned. The calcium NC-518® is claimed to be highly absorbable, particularly due to its formation, which is based on unique, patented technology. The technology involves both Zet Mill/Ball Mill bottom-up pulverizing technology and 4D top-down ebonite charging technology. In comparison to other technologies that use water and high heat, which results in a loss of core nutrients and decreased product efficacy, the calcium produced by this unique pulverizing technology is micron-sized (0.3um – 10um) and therefore more effective. While its tiny size significantly increases its absorption, it is the intense activation of zinc via the patented ebonite charging process that, the inventor claims, maximizes the bioavailability of this calcium in the body. In a published animal study (Asian-Aust. J. Anim. Sci. Vol. 20, No. 8: 1266 – 1271 August 2007), NC518® sharply improved the bone mineral densities of ovariectomized rats. This study was conducted to prove or disprove whether a similar increase in bone mineral density is possible in humans. Purpose In this study, we analyze the effect of orally administered NC-518® calcium on the bone mineral densities of men and post-menopausal women with osteoporosis or osteopenia. Study Material NC-518® is a combination of calcium and vitamin D3. Each capsule consists of 400mg of pulverized calcium and 400IU of vitamin D3. It is derived from oyster shells, with the calcium carbonate extracted during the pulverization process. The placebo arm consists of popular calcium carbonate with vitamin D3 at doses of 600mg of calcium and 400IU of vitamin D3 per caplet. Study Population The subjects consist of men and postmenopausal women over the age of 50 who have bone mineral density T-scores of less than -1.

Study Design This is a double blind, placebo controlled study in which each participant was randomly given either NC518® or Calcium Carbonate. NC-518® is made up of 400mg of NC-518® calcium and 400IU of vitamin D3. Calcium Carbonate consists of 600mg of calcium carbonate and 400IU of vitamin D3. The study distinguished two groups of participants. Both groups were instructed to take 2 capsules or caplets a day. One group consisted of participants who were not taking bisphosphonates (Fosamax® , Actonel® , or Boniva® ) and was called the A Group. The other group consisted of participants who were on bisphosphonates and was called the B Group. The target numbers for the study were 30 for the nonbisphosphonate A Group and 20 for the bisphosphonate B Group. Of the 30 participants enrolled in the A group, 15 randomly received NC-518® calcium and the other 15 received Calcium Carbonate. Of the 20 participants in the bisphosphonate B group, 10 randomly received NC-518® calcium and the other 10 received Calcium Carbonate. A total of 50 participants were targeted to complete the study for analysis. All participants were first screened by physicians and evaluated to determine their eligibility. Once a participant was qualified, he or she had a baseline bone mineral density test (DEXA Scan) to determine if osteoporosis or osteopenia was present. Osteoporosis is defined as a T-score of less than -2.5, and osteopenia is defined as a T-score of greater than -2.5 but less than -1.0. Participants were eligible for the study if their T-score was less than -1.0. Participants with osteoporosis or osteopenia who were on bisphosphonates had to have been on the bisphosphonate for more than 6 months, and they continued their treatment while in this study. If the participants were already on conventional calcium therapy, they were asked to stop their current calcium therapy and were randomly assigned to either NC-518® or Calcium Carbonate. The participants not on calcium supplementation were randomly assigned to either NC-518® or Calcium Carbonate. DEXA Scan measurements were done at 0 and 4 months to assess and compare the efficacy of NC518® calcium on osteoporosis and osteopenia to the efficacy of Calcium Carbonate. The rationale for conducting DEXA Scan measurements at 4 months was that it takes bones about four months for absorption and for the construction of new bones. Bone Mineral Density Testing Bone mineral density was measured using the GE Lunar Prodigy system. System calibration and quality assurance was done per manufacturer guidelines. Test Period The recruitment period of 6 months started on March 1, 2010 and ended on September 1, 2010. The subjects had baseline bone density testing. Once their qualification for the study was established and approved by the principal investigator, the participants were randomly assigned to take NC-518® or Calcium Carbonate for a 4 month period. Bone mineral density testing was repeated after 4 months of taking the provided calcium. The study is expected to be completed on January 1, 2011 or earlier, depending on the recruitment process. Study Procedure VISIT 1: 1. All participants have the risks and benefits explained by the principal investigator. 2. All participants have ample time to ask any questions about the study. 3. The principal investigator reviews the inclusion and exclusion criteria of each participant. 4. Once the inclusion and exclusion criteria have been met, all participants have the DEXA Scan performed. 5. Once qualified, each participant is given a bottle containing either NC-518® or Calcium Carbonate. 6. All participants are instructed to take the pills, one tablet two times a day after a meal.

7. All participants are instructed to return in 4 months to repeat the DEXA Scan measurement. 8. All participants are instructed not to change their lifestyles, including diet and exercise routines. VISIT 2: 1. All participants have their second DEXA Scan measurements. 2. Upon completion of the study at 4 months, all participants are instructed to resume their previous calcium therapies. 3. Any adverse reactions to the pills are asked and recorded.

Primary End Point The primary end point of the study is the improvement of bone mineral density as assessed by the DEXA Scan.

Data Of the 53 participants enrolled in the study, 49 participants completed the study in compliance with the study protocol. These 49 participants are included in the final analysis. Of the 4 participants excluded from the study, 1 belonged to the non-bisphosphonate group and 3 belonged to the bisphosphonate group. In the non-bisphosphonate group, 1 participant failed to return for the second DEXA Scan. In the bisphosphonate group, 2 participants failed to consume at least 50% of the pills supplied, and 1 participant failed to return for the second DEXA Scan. Of those who successfully completed the study, 30 participants belonged to the non-bisphosphonate group and 19 participants belonged to the bisphosphonate group. The average age of the participants was 66. The average age of the participants was 68 for the NC-518® group and 63 for the Calcium Carbonate group. A breakdown of the participants in the each group is illustrated in Figure 1 below. Figure 1. Breakdown of participants in subgroups.

Of the 25 participants in the NC-518® group, 16 participants showed improvement in their bone density measurements, 6 showed no change, and 3 showed worsening. Of the 24 participants in the Calcium Carbonate group, 9 participants showed improvement in their bone density measurements, 4 showed no change, and 11 showed worsening. This is shown in Table 1. Participants in the NC-518® group showed statistically significant improvement in their DEXA Scan T-scores compared to those in the Calcium Carbonate group (p=0.009).

Table 1. Both Bisphosphonate and Non-Bisphosphonate Groups Combined IMPROVED

UNCHANGED

WORSENED

NC-518® (25)

16

6

3

CALCIUM CARBONATE(24)

9

4

11

The NC-518® and Calcium Carbonate groups are further divided into non-bisphosphonate and bisphosphonate groups. The non-bisphosphonate group consisted of 30 participants; 15 participants belonged to the NC-518® group and the other 15 belonged to the Calcium Carbonate group. Of the 15 participants in the nonbisphosphonate group who took NC-518® , 9 participants showed improvement in their bone density measurements, 3 showed no change, and 3 showed worsening. Of the 15 participants who took Calcium Carbonate, 5 showed improvement in their bone density measurements, 3 showed no change, and 7 showed worsening. This is shown in Table 2. The average T-score improvement in the NC-518® group was +0.16, while in the Calcium Carbonate group, T-scores were decreased by -0.11. This is shown in Table 3. More participants in the NC-518® group showed improvement in bone mineral density than did those of the Calcium Carbonate group. The average improvement in T-scores was significantly higher in the NC-518® group. This was statistically significant (p=0.041).

Table 2. Non-bisphosphonate Group: Comparison of groups that consumed NC-518® vs. Calcium Carbonate IMPROVED

UNCHANGED

WORSENED

NC-518® (15)

9

3

3

CALCIUM CARBONATE(15)

5

3

7

Table 3. Non-bisphosphonate Group: Final T-score and average T-score changes before and after NC518® and Placebo (Calcium Carbonate)

NC-518® Placebo/Calcium Carbonate

Subjects 15 15

Improve 9 5

Same 3 3

Worse 3 7

Ave. T Before -2.93 -2.1

Ave. T After -2.77 -2.21

Change +0.16 -0.11

The bisphosphonate group consisted of 19 participants; 10 participants belonged to the NC-518® group and the other 9 belonged to the Calcium Carbonate group. Of the 10 participants in the bisphosphonate group who consumed NC-518® , 7 participants showed improvement in the bone density measurements, 3 showed no change, and none showed worsening. Of the 9 participants who took Calcium Carbonate, 4 showed improvement in their bone mineral densities, 1 showed no change, and 4 showed worsening. This is shown in Table 4. The average T-score improvement in the NC-518® group was +0.19, while in the Calcium Carbonate group, the average T-score improvement was 0.00. This is shown in Table 5. While the NC-518® group showed significant improvement compared to the Calcium Carbonate group, it was not statistically significant (p=0.111).

Table 4. Bisphosphonate Group: Comparison of groups that consumed NC-518® vs. Calcium Carbonate IMPROVED

UNCHANGED

WORSENED

NC-518® (10)

7

3

0

CALCIUM CARBONATE(9)

4

1

4

Table 5. Bisphosphonate Group: Final T-score and average T-score changes before and after NC-518® and Placebo (Calcium Carbonate): A: Actonnel® (9), B: Boniva® (6), F: Fosamax® (4)

NC-518® Placebo/Calcium Carbonate

Subjects 10

Improve 7

Same 3

Worse 0

Ave. T Before -2.77

Ave. T After -2.58

Change +0.19

9

4

1

4

-2.27

-2.27

0.00

Discussion In this double blind, placebo controlled study involving 49 participants who were randomly assigned to either NC-518® or popular Calcium Carbonate, those who consumed NC-518® showed statistically significant improvement in their bone density measurements. (p=0.009) In the non-bisphosphonate group, after 4 months of pills, the NC-518® participants showed statistically significant improvement in their T-scores (p=0.041). The NC-518® group showed an average T-score improvement of +0.16 (about a 2% improvement). A 1 standard deviation difference in T-scores equals about a 10-15% difference in bone mineral densities. This corresponds to the approximately 2% improvement in the bone density scores of participants who consumed NC-518® over the 4 month period. In the bisphosphonate group, the NC-518® group showed an average T-score improvement of +0.19, while the Calcium Carbonate group showed no change. Although the participants in the NC-518® group showed greater improvement in their bone mineral density measurements, it was not statistically significant (p value = 0.111). In the participants who were already on bisphosphonates, the addition of NC-518® improved their bone density scores by about 2.3% in the 4 month period, whereas the Calcium Carbonate group showed no change in bone mineral density T-scores. Bisphosphonates are widely used to prevent and treat osteoporosis. The bisphosphonates used in this study (Fosamax® , Actonel® , and Boniva® ) are nitrogenous bisphosphonates, which counter the action of osteoclasts by preventing bone breakdown. In the non-bisphosphonate group of this study, in which participants only consumed either NC-518® or Calcium Carbonate, NC-518® showed statistically significant improvement in bone mineral density. This supports the hypothesis that NC-518® alone can improve bone mineral density. In the bisphosphonate group, the addition of NC-518® improved bone mineral density but was not statistically significant. This can be attributed to the fact that bisphosphonates play a role in the remodeling process of bones and potentially have positive effect on bone mineral density. Maximum improvement was seen in the participants who consumed both bisphosphonates and NC-518® , while most statistically significant improvement was seen in the participants who consumed NC-518® alone.

Conclusion The significant improvement in T-scores in those who took NC-518® rather than Calcium Carbonate is believed to be due to NC-518®’s better absorption and bioavailability. In the published animal study by H.S. Park et al., which compared NC-518® to ionized calcium, calcium carbonate, and no calcium supplementation in ovariectomized rats, the rats treated with NC-518® showed statistically significant 3 improvement in bone mineral density and trabecular bone area compared to the other three groups. This improvement in the bone mineral densities of ovariectomized rats was replicated in this current study, where statistical improvement was seen in the bone mineral densities of persons with osteopenia and osteoporosis. Osteoporosis is a serious medical condition with high mortality and morbidity rates. It involves complex interactions on both the hormonal and cellular levels. Calcium and vitamin D play essential roles in building and maintaining bone metabolism. While there are a number of medical treatments available to combat this potentially life threatening condition, proper exercise and the consumption of vitamins and minerals, particularly calcium and vitamin D, are essential for its prevention and treatment. Although bone mineral density worsens with age, especially in the postmenopausal state due to depressed hormonal support, this study showed that NC-518® results in statistically significant improvement in bone density scores.

References 1. 2. 3.

National Osteoporosis Foundation; http://www.nof.org/node/40, 2010 Donna Staton, Marcu Harding, Faqs.org, Nutrition & Well-Being A to Z, Ca-De, Calclium H.S. Park, B.J. Jeon, J. Ahn, H.S. Kwak, Effect of Nanocalcium Supplemented Milk on Bone Calcium Metabolism in Ovariectomized Rates, Asian-Aust. J. Anim. Sci., Vol. 20, No. 8: 12661271, August 2007