Endothelial Dysfunction, Obesity, Vitamin D Deficiency
Conditions
Keywords
Obesity, Endothelial dysfunction, Vitamin D deficiency
Brief summary
Vitamin D deficiency has been linked to endothelial dysfunction in adults. Obese adolescents have a high prevalence of Vitamin D deficiency as well as evidence of endothelial dysfunction. Our hypothesis is that supplementation of Vitamin D deficient adolescents with Vitamin D would lead to improvement in endothelial dysfunction.
Detailed description
Subjects had a brief screening visit with one of the study team members. Past medical history, current medications and a brief dietary history was taken. The dietary history was taken to assess calcium intake in a day and was obtained via the validated Short Calcium questionnaire. Blood pressure, heart rate as well as height,weight, waist and hip circumference measurement was obtained by study staff. In addition, a brief physical examination to determine Tanner stage was also completed. This involved examination of both breasts and genitalia for adolescent female and only genitalia in males. A physical activity questionnaire (IPAQ) was also administered by the study staff at this visit. A blood draw of 5 ml to measure plasma 25(OH) D, calcium, phosphorus was obtained as part of the screening visit. For girls who had started menstruating, a urine pregnancy test was also obtained. After the screening visit eligible subjects had baseline biochemical tests and an endothelial function assessment. Study participants received a pill container with six total pills of vitamin cholecalciferol (D3) (1 pill = 50,000 IU), with directions to take two pills once a month (100,000 IU) at the same time for a period of 3 months. Compliance was assessed at the 3 month visit by counting the number of pills remaining in the container. At 1 month, the subjects had tests for serum calcium and 25 hydroxy vitamin D levels. A urine test for random calcium to creatinine ratio was also obtained. At 2 months, the serum calcium, 25 hydroxy vitamin D levels, and calcium to creatinine ratio tests were repeated. At the end of 3 months, biochemical tests and an endothelial function assessment were repeated. Medical history, questionnaires, and the physical exam were also repeated.
Interventions
DIETARY_SUPPLEMENTVitamin D3
Vitamin D 3 at 100,000 IU once a month for 3 months
Sponsors
Mayo Clinic
Study design
Allocation
NA
Intervention model
SINGLE_GROUP
Primary purpose
PREVENTION
Masking
NONE
Eligibility
Sex/Gender
ALL
Age
12 Years to 18 Years
Healthy volunteers
Yes
Inclusion criteria
1. Age 12-18 years 2. BMI \>95% for age and gender 3. 25 (OH) D levels less than 30 ng/ml
Exclusion criteria
1. 25 (OH) D levels \>30 ng/mL 2. Serum calcium \>10.4 mg/dL 3. Serum phosphorus \> 4.7 mg/dl 4. Pregnancy or nursing 5. Current cancer 6. Patients on vitamin D3 supplementation exceeding 400 IU/day 7. Hypertension defined as Blood Pressure over the 95th percentile for age, gender and height 8. Dietary calcium intake exceeding 1500 mg/day 9. Hepatic or renal disorders 10. Type 1 or type 2 diabetes mellitus 11. Subjects receiving insulin, metformin, or oral hypoglycemic medications 12. Subjects with malabsorption disorders (celiac disease, cystic fibrosis, inflammatory bowel disease)
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Flow Mediated Dilatation (FMD) | baseline, 3 months | Endothelial function was assessed by FMD, via a high-resolution Doppler ultrasonography examination of the right brachial artery. FMD was calculated as the maximal percentage increase in brachial artery diameter (BAD) from baseline after the release of cuff occlusion. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Total Cholesterol | baseline, 3 months | Total cholesterol levels were measured by an enzymatic colorimetric assay. |
| Triglycerides | baseline, 3 months | Total triglyceride levels were measured by an enzymatic colorimetric assay. |
| Body Mass Index | baseline, 3 months | Body Mass Index (BMI) is a health index for comparing weight to height. BMI is a person's weight in kilograms (kg) divided by his or her height in meters squared. The body mass index is an indication if a person is at a suitable weight for his height on an approximation of body fat. A body mass index of under 20 is considered to be underweight, while a body mass index between 20 to 25 is considered healthy. A body mass index in the range of 25 to 30 is regarded as overweight. A body mass index over 30 is regarded as obese. |
| International Physical Activity Questionnaire (IPAQ) Short Form Score | baseline, 3 months | The IPAQ short form used asked 7 questions about activities in the last 7 days, covering vigorous physical activities, moderate activities, walking, and sitting, asking for days per week, hours per day or minutes per day. The score is reported in metabolic equivalent (MET)-minutes per week. Possible scores could range from 0 (inactive) to greater than 3000 MET-minutes/week (highly active). The definition of high activity was vigorous intensity activity on at least 3 days achieving a minimum total activity of at least 1500 MET-minutes/week OR 7 days of any combination of walking, moderate-intensity or vigorous-intensity activities achieving a minimum total physical activity of at least 3000 MET-minutes/week. Therefore a score of \> 3000 MET-minutes/week was possible. |
| Calcium Intake Per Day | baseline, 3 months | Calcium intake was measured using the validated Short Calcium Questionnaire (SCQ). This questionnaire is in the form of an spreadsheet, and asks the participant to enter the number of servings per week of various food items and vitamin or mineral supplements. The spreadsheet calculates the daily calcium intake (mg/day) from the data entered. |
| Serum Parathyroid Hormone (PTH) | baseline, 3 months | A parathyroid hormone (PTH) blood test measures the level of parathyroid hormone in the blood. This test is used to help identify hyperparathyroidism, to find the cause of abnormal calcium levels, or to check the status of chronic kidney disease. PTH controls calcium and phosphorus levels in the blood. PTH was measured by a two-site chemiluminescent immunometric assay. |
| Fasting Glucose | baseline, 3 months | Plasma glucose was measured by hexokinase enzymatic assay. |
| 25-hydroxy Vitamin D (25[OH]D) Levels | baseline, 3 months | 25(OH)D was measured using liquid chromatography-tandem mass spectrometry. Total 25(OH)D concentrations of each sample was calculated using internal standard, 25(OH)D\_2 and 25(OH)D\_3. |
| Homeostatic Model Assessment of Insulin Resistance Index (HOMA-IR) | baseline, 3 months | This calculation measures insulin resistance, and requires U.S. standard units. The healthy range is 0.5 to 1.4. Less than 1.0 means the subject is insulin-sensitive, which is optimal. Above 1.9 indicates early insulin resistance. Above 2.9 indicates significant insulin resistance. The HOMA-IR was calculated as: HOMA-IR = fasting serum glucose (mmol/L) x fasting insulin (mU/mL)/22.5. |
| High Sensitivity C-reactive Protein (Hs-CRP) | baseline, 3 months | A high-sensitivity C-reactive protein (hs-CRP) test may be used to help evaluate an individual for risk of cardiovascular disease (CVD). C-reactive protein (CRP) is a protein that increases in the blood with inflammation. Studies have suggested that a persistent low level of inflammation plays a major role in atherosclerosis, the narrowing of blood vessels due to build-up of cholesterol and other lipids, which is often associated with CVD. The hs-CRP test accurately measures low levels of C-reactive protein to identify low but persistent levels of inflammation and thus helps predict a person's risk of developing CVD. hs-CRP was measured using particle-enhanced immunonephelometry. |
| Low-density Lipoprotein Cholesterol (LDL) Cholesterol Levels | baseline, 3 months | The test for low-density lipoprotein cholesterol is used as part of a lipid profile to predict an individual's risk of developing heart disease. A desirable level is \<3.36 mmol/L; borderline high is 3.36 - 4.11 mmol/L; high is \>/= 4.14 mmol/L. LDL cholesterol was calculated as: LDL = Total cholesterol - HDL cholesterol - Triglycerides/5. |
| High Density Lipoprotein (HDL) Cholesterol Levels | baseline, 3 months | Total HDL cholesterol levels were measured by an enzymatic colorimetric assay. The test for high-density lipoprotein cholesterol (HDL-C) is used along with other lipid tests to screen for unhealthy levels of lipids and to determine the risk of developing heart disease. If a subject has a negative risk factor, a desirable HDL level would be \>/= 1.55 mmol/L. |
| Urine Calcium to Creatinine Ratio | baseline, 3 months | Urine calcium/creatinine ratio (unit mg/g) on random urine sample was calculated by dividing calcium in mg by creatinine in g. |
| Reactive Hyperemia Index (RHI) | baseline, 3 months | The cuff of a sphygmomanometer was placed on the forearm and inflated to 50 mm Hg above the participant's systolic blood pressure for a period of 5 min. The increase in resting brachial blood flow was calculated as the maximum flow recorded in the first 15 seconds after cuff deflation and expressed as a percentage increase from baseline reactive. Higher values are considered normal or improved endothelial function. |
| Fasting Insulin | baseline, 3 months | Serum insulin was measured using commercial electrochemiluminescence immunoassay kits. |
Countries
United States
Participant flow
Recruitment details
Subjects were recruited between February 2013 and December 2013 at Mayo Clinic in Rochester, Minnesota.
Participants by arm
| Arm | Count |
|---|---|
| Vitamin D3 Vitamin D3 supplementation at 100,000 IU once a month for 3 months. | 19 |
| Total | 19 |
Baseline characteristics
| Characteristic | Vitamin D3 |
|---|---|
| Age, Continuous | 15.8 years STANDARD_DEVIATION 1.7 |
| Region of Enrollment United States | 19 participants |
| Sex: Female, Male Female | 7 Participants |
| Sex: Female, Male Male | 12 Participants |
Adverse events
| Event type | EG000 affected / at risk |
|---|---|
| deaths Total, all-cause mortality | — / — |
| other Total, other adverse events | 0 / 19 |
| serious Total, serious adverse events | 0 / 19 |
Outcome results
Primary
Flow Mediated Dilatation (FMD)
Endothelial function was assessed by FMD, via a high-resolution Doppler ultrasonography examination of the right brachial artery. FMD was calculated as the maximal percentage increase in brachial artery diameter (BAD) from baseline after the release of cuff occlusion.
Time frame: baseline, 3 months
Population: Data for one subject could not be included as the data on FMD was lost in the system and could not be retrieved.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Flow Mediated Dilatation (FMD) | Baseline FMD | 9.5 percentage increase in BAD | Standard Deviation 3.52 |
| Vitamin D3 | Flow Mediated Dilatation (FMD) | 3 months FMD | 10.4 percentage increase in BAD | Standard Deviation 3.82 |
Comparison: P-value for change from baseline to 3 month follow-up.p-value: 0.59Wilcoxon (Mann-Whitney)
Secondary
25-hydroxy Vitamin D (25[OH]D) Levels
25(OH)D was measured using liquid chromatography-tandem mass spectrometry. Total 25(OH)D concentrations of each sample was calculated using internal standard, 25(OH)D\_2 and 25(OH)D\_3.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | 25-hydroxy Vitamin D (25[OH]D) Levels | baseline 25(OH)D | 55.9 nmol/L | Standard Deviation 12.2 |
| Vitamin D3 | 25-hydroxy Vitamin D (25[OH]D) Levels | 3 months 25(OH)D | 86.9 nmol/L | Standard Deviation 16.7 |
Comparison: Change from baseline to 3 month follow up visit.p-value: <0.001Wilcoxon (Mann-Whitney)
Secondary
Body Mass Index
Body Mass Index (BMI) is a health index for comparing weight to height. BMI is a person's weight in kilograms (kg) divided by his or her height in meters squared. The body mass index is an indication if a person is at a suitable weight for his height on an approximation of body fat. A body mass index of under 20 is considered to be underweight, while a body mass index between 20 to 25 is considered healthy. A body mass index in the range of 25 to 30 is regarded as overweight. A body mass index over 30 is regarded as obese.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Body Mass Index | baseline | 36.1 kg/m^2 | Standard Deviation 6.03 |
| Vitamin D3 | Body Mass Index | 3 months | 36.4 kg/m^2 | Standard Deviation 6.11 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.32Wilcoxon (Mann-Whitney)
Secondary
Calcium Intake Per Day
Calcium intake was measured using the validated Short Calcium Questionnaire (SCQ). This questionnaire is in the form of an spreadsheet, and asks the participant to enter the number of servings per week of various food items and vitamin or mineral supplements. The spreadsheet calculates the daily calcium intake (mg/day) from the data entered.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Calcium Intake Per Day | baseline | 1102.7 mg/day | Standard Deviation 304.3 |
| Vitamin D3 | Calcium Intake Per Day | 3 months | 975.5 mg/day | Standard Deviation 282.2 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.02Wilcoxon (Mann-Whitney)
Secondary
Fasting Glucose
Plasma glucose was measured by hexokinase enzymatic assay.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Fasting Glucose | baseline | 4.9 mmol/L | Standard Deviation 0.28 |
| Vitamin D3 | Fasting Glucose | 3 months | 4.97 mmol/L | Standard Deviation 0.26 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.09Wilcoxon (Mann-Whitney)
Secondary
Fasting Insulin
Serum insulin was measured using commercial electrochemiluminescence immunoassay kits.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Fasting Insulin | baseline | 225.71 pmol/L | Standard Deviation 127.93 |
| Vitamin D3 | Fasting Insulin | 3 months | 245.16 pmol/L | Standard Deviation 150.01 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.27Wilcoxon (Mann-Whitney)
Secondary
High Density Lipoprotein (HDL) Cholesterol Levels
Total HDL cholesterol levels were measured by an enzymatic colorimetric assay. The test for high-density lipoprotein cholesterol (HDL-C) is used along with other lipid tests to screen for unhealthy levels of lipids and to determine the risk of developing heart disease. If a subject has a negative risk factor, a desirable HDL level would be \>/= 1.55 mmol/L.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | High Density Lipoprotein (HDL) Cholesterol Levels | baseline | 1.2 mmol/L | Standard Deviation 0.28 |
| Vitamin D3 | High Density Lipoprotein (HDL) Cholesterol Levels | 3 months | 1.16 mmol/L | Standard Deviation 0.23 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.41Wilcoxon (Mann-Whitney)
Secondary
High Sensitivity C-reactive Protein (Hs-CRP)
A high-sensitivity C-reactive protein (hs-CRP) test may be used to help evaluate an individual for risk of cardiovascular disease (CVD). C-reactive protein (CRP) is a protein that increases in the blood with inflammation. Studies have suggested that a persistent low level of inflammation plays a major role in atherosclerosis, the narrowing of blood vessels due to build-up of cholesterol and other lipids, which is often associated with CVD. The hs-CRP test accurately measures low levels of C-reactive protein to identify low but persistent levels of inflammation and thus helps predict a person's risk of developing CVD. hs-CRP was measured using particle-enhanced immunonephelometry.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | High Sensitivity C-reactive Protein (Hs-CRP) | baseline | 47.62 nmol/L | Standard Deviation 35.43 |
| Vitamin D3 | High Sensitivity C-reactive Protein (Hs-CRP) | 3 months | 40 nmol/L | Standard Deviation 25.7 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.33Wilcoxon (Mann-Whitney)
Secondary
Homeostatic Model Assessment of Insulin Resistance Index (HOMA-IR)
This calculation measures insulin resistance, and requires U.S. standard units. The healthy range is 0.5 to 1.4. Less than 1.0 means the subject is insulin-sensitive, which is optimal. Above 1.9 indicates early insulin resistance. Above 2.9 indicates significant insulin resistance. The HOMA-IR was calculated as: HOMA-IR = fasting serum glucose (mmol/L) x fasting insulin (mU/mL)/22.5.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Homeostatic Model Assessment of Insulin Resistance Index (HOMA-IR) | baseline | 7.11 index of beta cell function | Standard Deviation 4.18 |
| Vitamin D3 | Homeostatic Model Assessment of Insulin Resistance Index (HOMA-IR) | 3 months | 7.9 index of beta cell function | Standard Deviation 5.1 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.18Wilcoxon (Mann-Whitney)
Secondary
International Physical Activity Questionnaire (IPAQ) Short Form Score
The IPAQ short form used asked 7 questions about activities in the last 7 days, covering vigorous physical activities, moderate activities, walking, and sitting, asking for days per week, hours per day or minutes per day. The score is reported in metabolic equivalent (MET)-minutes per week. Possible scores could range from 0 (inactive) to greater than 3000 MET-minutes/week (highly active). The definition of high activity was vigorous intensity activity on at least 3 days achieving a minimum total activity of at least 1500 MET-minutes/week OR 7 days of any combination of walking, moderate-intensity or vigorous-intensity activities achieving a minimum total physical activity of at least 3000 MET-minutes/week. Therefore a score of \> 3000 MET-minutes/week was possible.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | International Physical Activity Questionnaire (IPAQ) Short Form Score | baseline | 1786.6 MET-minutes per week | Standard Deviation 1506.2 |
| Vitamin D3 | International Physical Activity Questionnaire (IPAQ) Short Form Score | 3 months | 2799.1 MET-minutes per week | Standard Deviation 3834.6 |
Comparison: Change from baseline to 3-month follow-up.p-value: 0.21Wilcoxon (Mann-Whitney)
Secondary
Low-density Lipoprotein Cholesterol (LDL) Cholesterol Levels
The test for low-density lipoprotein cholesterol is used as part of a lipid profile to predict an individual's risk of developing heart disease. A desirable level is \<3.36 mmol/L; borderline high is 3.36 - 4.11 mmol/L; high is \>/= 4.14 mmol/L. LDL cholesterol was calculated as: LDL = Total cholesterol - HDL cholesterol - Triglycerides/5.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Low-density Lipoprotein Cholesterol (LDL) Cholesterol Levels | baseline | 1.95 mmol/L | Standard Deviation 0.66 |
| Vitamin D3 | Low-density Lipoprotein Cholesterol (LDL) Cholesterol Levels | 3 months | 2.15 mmol/L | Standard Deviation 0.81 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.05Wilcoxon (Mann-Whitney)
Secondary
Reactive Hyperemia Index (RHI)
The cuff of a sphygmomanometer was placed on the forearm and inflated to 50 mm Hg above the participant's systolic blood pressure for a period of 5 min. The increase in resting brachial blood flow was calculated as the maximum flow recorded in the first 15 seconds after cuff deflation and expressed as a percentage increase from baseline reactive. Higher values are considered normal or improved endothelial function.
Time frame: baseline, 3 months
Population: Data for one subject could not be included as the data on RHI was lost in the system and could not be retrieved.
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Reactive Hyperemia Index (RHI) | baseline | 449.3 percentage increase in blood flow | Standard Deviation 243.5 |
| Vitamin D3 | Reactive Hyperemia Index (RHI) | 3 months | 513.9 percentage increase in blood flow | Standard Deviation 325.6 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.47Wilcoxon (Mann-Whitney)
Secondary
Serum Parathyroid Hormone (PTH)
A parathyroid hormone (PTH) blood test measures the level of parathyroid hormone in the blood. This test is used to help identify hyperparathyroidism, to find the cause of abnormal calcium levels, or to check the status of chronic kidney disease. PTH controls calcium and phosphorus levels in the blood. PTH was measured by a two-site chemiluminescent immunometric assay.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Serum Parathyroid Hormone (PTH) | baseline | 4.1 pmol/L | Standard Deviation 1.6 |
| Vitamin D3 | Serum Parathyroid Hormone (PTH) | 3 months | 3.3 pmol/L | Standard Deviation 1.1 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.0123Wilcoxon (Mann-Whitney)
Secondary
Total Cholesterol
Total cholesterol levels were measured by an enzymatic colorimetric assay.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Total Cholesterol | baseline | 3.69 mmol/L | Standard Deviation 0.71 |
| Vitamin D3 | Total Cholesterol | 3 months | 4.03 mmol/L | Standard Deviation 0.87 |
Comparison: Change from baseline to 3 month follow-up.p-value: <0.01Wilcoxon (Mann-Whitney)
Secondary
Triglycerides
Total triglyceride levels were measured by an enzymatic colorimetric assay.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Triglycerides | Baseline | 1.19 mmol/L | Standard Deviation 0.57 |
| Vitamin D3 | Triglycerides | 3 months | 1.58 mmol/L | Standard Deviation 0.81 |
Secondary
Urine Calcium to Creatinine Ratio
Urine calcium/creatinine ratio (unit mg/g) on random urine sample was calculated by dividing calcium in mg by creatinine in g.
Time frame: baseline, 3 months
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Vitamin D3 | Urine Calcium to Creatinine Ratio | baseline | 67.8 mg/g | Standard Deviation 59.2 |
| Vitamin D3 | Urine Calcium to Creatinine Ratio | 3 months | 87.7 mg/g | Standard Deviation 73.2 |
Comparison: Change from baseline to 3 month follow-up.p-value: 0.32Wilcoxon (Mann-Whitney)