Evaluation of a Milk-Based Nutritional Supplement to Modify Diurnal Rates of Bone Turnover and Effect a Positive Change in Bone Health in Post-Menopausal Women at Risk of Osteoporosis
Phase: NA
Conditions: Bone and Bones, Osteoporosis Risk, Osteoporosis, Postmenopausal
Sponsor: University of Limerick

Nutritional Supplement and Bone Health in Post-Menopausal Women (MBPS) 
Ethical approval number: 2017_06_03_EHS                                        Date of Approval 26th June 2017 
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Study Design and Protocol 
 
An Evaluation of a Milk-Based Nutritional Supplement to Effect a Positive 
Change in Bone Health in Post-Menopausal Women at Risk of Osteoporosis. 
 
In a novel approach to the timing of nutrient ingestion, the proposed nutrient intervention 
seeks to modify (reduce) the rate of bone resorption and promote the rate of bone 
formation to the benefit of bone health in this at risk population. 
  
2. Study design: A block randomised, cross-over design of 24h rates of bone turnover in 
healthy, post-menopausal women with osteopenia receiving either a milk-based protein 
supplement (MBPS) or isoenergetic (maltodextran) placebo control (PLACEBO). 
3. Procedures:   
Subjects: Post-menopausal women with osteopenia as determined by site-specific BMD 
(DXA) diagnosed and screened by a clinician (Dr Manjula Hettiarachchi) and for dietary 
intake of calcium and Vit D by a clinical dietitian (Dr Catherine Norton).  
Subject screening (clinical examination, 1h) and dietary intake of calcium and Vit D (by 
food frequency questionnaire, 1h) will precede the experimental protocol. 
  
Experimental protocol and data collection: Supplemenary Material contains a Flowchart 
Subjects will attend for a 2 day overnight residence in the National Altitude Training 
Centre, Kilmurray Village. This facility is equipped to conduct residential human trials and 
has on-site individual bedroom accommodation subjects & researchers, a living room with 
fully equipped kitchen and laboratory to handle human blood samples.  
The subjects' programmed protocol is as follows; 
1. Arrive @ 17:00h with overnight bag;  
2. Empty bladder and then provide and retain urine samples for the duration of 
the stay (assisted collection by researchers); 
3. Consume a standardised evening meal (pre-prepared by the research 
dietitian) and then relax reading/watching films etc in the living room;  
4. At 20:00h a research nurse will insert a cannula into a superficial arm vein 
and a blood draw (5ml) will be taken and processed for later analysis; 
5. Further blood draws (5ml) will be taken at 22:00h, 2300h, 2400h,0100h and 
0200h and the cannula withdrawn; 
6. At 22:00h consume either maltodetran control (PLACEBO) (day 1) OR 
supplement (MBPM)(day 2)  - or vice versa; 
7. Retire to bedroom; 
8. Consume a standardised breakfast and lunch (pre-prepared by the research 
dietitian) whilst living in and around the University grounds  
     (i.e. in close proximity to ensure 24h urine collection is complete); 
9. Repeat from 2 above to end of 2nd day. 
  
Total blood draw over 48h is 14 x 5ml = 70ml 
All formulations to be supplied food grade and product tested by Dairygold Co-
opertaive Society, Mitchelstown, Ireland. 
  
4. Analytical methods: 
i. 
In total 224 (16 subjects * 7 samples * 2 d) blood samples will be collected. 
Separated aliquots will be frozen @ -80C and batch analysed for CTX (biomarker of 
resorption), P1NP (biomarker of formation). In total 32 (16 subjects * 2 d) 24h urine 
samples will be collected. Aliquots will be frozen @ -80C and batch analysed for 
uPYD, uDPD (biomarkers of resorption) normalised to urinary creatinine.  

Nutritional Supplement and Bone Health in Post-Menopausal Women (MBPS) 
Ethical approval number: 2017_06_03_EHS                                        Date of Approval 26th June 2017 
2 | P a g e  
 
ii. The I.O.F. recommended serum marker for bone resorption is CTX and formation 
P1NP. The magnitude and time course of the acute response (0-4h) change in bone 
turnover following ingestion of the MBPM vs. PLACEBO will be analysed by repeated 
measures ANOVA of the serum biomarkers of resorption and formation. 
iii.  The overall influence on the diurnal change in bone turnover is best measured by 
a global biomarker of bone resorption specific to the type I collagen in bone. This 
is provided by the measurement of 24h urinary pyridinoline (PYD) and 
deoxypyridinoline (DPD) excretion normalised for urinary creatinine. Thus, the 
magnitude of effect of 24h change in bone resorption following ingestion of the 
MBPM vs. PLACEBO will be analysed by ANOVA(R) of the urinary markers of bone 
resorption (PYD/creatinine and DPD/creatinine). 
 
 
 
 
 
 
Schematic Flow Chart of theTrial 
 
 
 
 

Nutritional Supplement and Bone Health in Post-Menopausal Women (MBPS) 
Ethical approval number: 2017_06_03_EHS                                        Date of Approval 26th June 2017 
3 | P a g e  
 
 
Volunteer Information Sheet 
 
An Evaluation of a Milk-Based Nutritional Supplement to Effect a Positive 
Change in Bone Health in Post-Menopausal Women at Risk of Osteoporosis. 
 
Thank you for considering this research study.  
 
What is the project about? 
 
It is an attempt to use a milk-based, nutrient supplement to make a positive modification to bone 
health in post-menopausal women at risk of osteoporosis, i.e. diagnosed as osteopenic. 
  
FIVE key facts related to bone health 
1. Over the lifespan the tendency to poor bone health is influenced by your genetic make-up and 
environmental factors such as diet and physical activity.  
 
2. Poor bone health is characterised by a diminution to bone quality (i.e. the protein content of 
bone) and low bone mass (i.e. the mineral content of the bone).  
 
3. We now know that bone regulates both quality and mineral content through a coupled 
process of bone resorption (the dissolution of old bone) and bone formation (the formation 
of new bone), a process termed bone turnover. 
 
4. Bone turnover has an underlying 24 clock, greater rates of bone resorption occur at night than 
in the day, and 
 
5. Bone turnover is modulated by calcium, Vit D and protein intake. Calcium and vitamin D 
intake, as recommended by the International Osteoporosis Foundation (I.O.F.), can modify 
bone turnover to reduce potential development of poor bone health. Equally, appropriate 
protein intake can increase the to the protein content of bone in support bone health. 
 
In a novel application of the 5 key facts listed above, this study seeks to modify the rate of bone 
turnover by feeding a milk-based nutritional supplement at night, i.e. when the rate of bone turnover 
is highest. We propose a milk-based supplement because milk contains a natural source of calcium 
and protein. So, if you are post-menopausal, diagnosed as at risk of osteoporosis but otherwise in 
good health you may wish to consider taking part in this study. 
 
 
What will you have to do? 
Firstly, we will make every effort to explain fully all aspects of the study and allow you time to question 
and reflect on your participation.  If you are happy to proceed, you will be asked to sign a written, 
statement of consent. 

Nutritional Supplement and Bone Health in Post-Menopausal Women (MBPS) 
Ethical approval number: 2017_06_03_EHS                                        Date of Approval 26th June 2017 
4 | P a g e  
 
 
You will be consenting to the following; 
 
1. Screening:   
Following an overnight fast (from 10pm the previous evening) to attend the University of Limerick 
at a scheduled date between 7:30 and 9:00am to undergo a clinical examination, provide a blood 
sample, have a bone scan (DXA) and complete a dietary intake analysis. This will take 
approximately 2h to complete. You will be provided with a snack, tea/coffee before leaving. 
 
It will take a few days to process these screening data. Should the results indicate that you are 
a suitable candidate for this study you will be invited to proceed to participate in the 
intervention trial. 
 
2. Intervention: 
This is a 2-day, 2-night trial requiring you to be resident at the University for 48 hours. 
 
At a designated date we require you to undertake the following; 
 
Attend a University Residence located in Kilmurray Village. This facility is equipped to conduct 
residential human trials and has on-site individual bedroom accommodation and living room with 
fully equipped kitchen. There are 6 rooms, so you will be ‘sharing’ the house with 5 other 
participants for the 2 days. 
 
The programmed protocol is as follows; 
1. 
Arrive @ 17:00h with overnight bag;  
2. 
Settle into your residence before emptying your bladder. From this tie point and for 
the 48 hours that follow you will be required to provide urine samples for the duration of the 
stay (assisted collection by researchers); 
3. 
Consume a standardised evening meal (pre-prepared by the research dietitian) and 
then relax reading/watching films etc. in the living room;  
4. 
At 20:00h a research nurse will insert a cannula into a superficial arm vein and a blood 
draw (5ml) will be taken and processed for later analysis; 
5. At 22:00h consume either a carbohydrate-based drink (CON) (day 1) OR milk-based 
protein supplement (day 2)  - or vice versa and a further blood sample will be drawn; 
6. Retire to bedroom; 
7. Further blood draws (5ml) will be taken at 2300h, 2400h, 0100h and 0200h after which 
the cannula is withdrawn. This will require the researcher to enter your room during the 
night. Though blood can be drawn without waking, there may be occasions when the 
disturbance will wake you. 
8. 
In the morning, consume a standardised breakfast, then a lunch (both breakfast and 
lunch are pre-prepared by the research dietitian) whilst living in and around the University 
grounds  
     (i.e. in close proximity to ensure 24h urine collection is complete); 
9. 
Repeat from 2 above to end of 2nd day. 
 
AND THAT’s IT! 
 
 
 
 
 

Nutritional Supplement and Bone Health in Post-Menopausal Women (MBPS) 
Ethical approval number: 2017_06_03_EHS                                        Date of Approval 26th June 2017 
5 | P a g e  
 
What are the potential benefits to you? 
 
The benefits for partaking in this study are: 
o You will receive a comprehensive report of current bone mineral density, body composition 
and dietary intake of essential nutrients related to bone health. 
o You will be contributing to the evaluation of a novel, diet-based intervention to enable bone 
health in those whom, like you, are at risk of osteoporosis. 
 
What are the potential risks? 
 
You may be assured that the procedures to be employed have been used extensively by the 
researchers conducting this study and are generally well tolerated by participants.  
 
o There is minimal risk involved in the study.  The bone scan is attained by exposing you to a 
mild X-ray when lying on a bed, termed a DXA scan. This exposure is equivalent to 
approximately 1/30th of a normal X-ray which is ‘trivial’.  
 
o You may experience some discomfort during the blood draw. Blood will be drawn by a 
qualified research nurse or clinician. 
 
o The food supplement is based on milk, a naturally source dietary food. The study would NOT 
be suitable for a person who has a known allergy to dairy produce, e.g. lactose intolerant. 
 
What if I do not want to take part? 
 
You can discontinue your participation in the research study at any time and this will be dealt 
with in an unhesitating and confidential manner.  
 
What happens to the information? 
 
The information retrieved will be dealt with and handled in complete confidence. After the 
completion of the study, information will be kept electronically on the principal investigator’s 
password‐protected computer 
 
Who else is taking part? 
Other post-menopausal women at risk of poor bone health. 
 
What if something goes wrong? 
In the unlikely event that anything untoward occur the testing procedure will immediately cease and 
the PESS department emergency procedures will be followed. 
 
What happens at the end of the study? 
At the end of the study the information garnered will be used anonymously to present the results as 
a conference or academic journal communication and may result in the supplement being further 
evaluated over a longer period of time. 
Your personal details/information and data from the study will be held by the principal investigator 
for up to 7 years in a password‐protected computer at UL.   
Upon completion, a report containing the overall study outcome will be available to participants on 
written request to the Principal Investigator. 
 
 
 
 

Nutritional Supplement and Bone Health in Post-Menopausal Women (MBPS) 
Ethical approval number: 2017_06_03_EHS                                        Date of Approval 26th June 2017 
6 | P a g e  
 
What if I have more questions or do not understand something. 
If you do not understand any aspect of the study we would urge you to come forward to any of the 
researchers (see contact details below) and discuss any questions that you might have. 
 
It is important that participants feel completely at ease throughout the experiment. 
 
Will I receive payment for participating? 
Subjects receive an overnight allowance for the time committed to the study.  
The allowance is €108 per night. 
 
 
Project Investigator Contact Details:   
 
Principal Investigator:   Professor Phil Jakeman 
P1027, University of Limerick, 
Office Telephone: 061 202800 
Email: phil.jakeman@ul.ie    
 
Other investigators: 
Dr. Catherine Norton 
 
 
 
Email: catherine.norton@ul.ie  
 
 
 
If you have any concerns about this study and wish to contact someone independent, you may 
contact The EHS Research Ethics Contact Point of the Education and Health Sciences Research Ethics 
Committee, Room E1003, University of Limerick, Limerick. 
Tel: (061) 234101 / Email: ehsresearchethics@ul.ie 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

Nutritional Supplement and Bone Health in Post-Menopausal Women (MBPS) 
Ethical approval number: 2017_06_03_EHS                                        Date of Approval 26th June 2017 
7 | P a g e  
 
Informed Consent 
 
An Evaluation of a Milk-Based Nutritional Supplement to Effect a Positive 
Change in Bone Health in Post-Menopausal Women at Risk of Osteoporosis. 
 
Should you agree to participate in this study please read the statements below and if you agree to 
them, please sign the consent form. 
o I have read and understood the participant information sheet.   
o I understand what the project is about, and what the results will be used for. 
o I understand that what the researchers find out in this study may be shared with others but 
that my name will not be given to anyone in any written material developed.   
o I am fully aware of all of the procedures involving myself, and of any risks and benefits 
associated with the study.   
o I know that my participation is voluntary and that I can withdraw from the project at any stage 
without giving any reason. 
o I consent to the data obtained from the conduct of this project to be used, anonymously, for 
presentation and publication. 
 
I consent (or agree) to my involvement in this research project after agreeing to all the above 
statements. 
 
Name: (please print): __________________________ 
 
Signature: ___________________________________     Date: ____________ 
 
Investigator’s Signature _________________________     Date: _____________ 
 
This study has been approved by the ethics committee of the Faculty of Education and Health Sciences.  
If you have any concerns about this study and wish to contact someone independent, you may contact The EHS 
Research Ethics Contact Point of the Education and Health Sciences Research Ethics Committee, University of 
Limerick, Limerick. Tel: (061) 234101 / Email: ehsresearchethics@ul.ie