Dietary Protein and Bone Health Emphasis: Animal Protein

USDA-ARS Human Nutrition Research Centers Dietary Protein and Bone Health Emphasis: Animal Protein Z.K. (Fariba) Roughead, PhD, RD Research Nutritionist USDA-ARS Grand Forks Human Nutrition Research Center 15 Senior Scientists 135 support staff 2 metabolic kitchens Service labs: clinical lab, mineral analysis 2 DXAs, Whole body counter

Osteoporosis affects 2 million worldwide 2% of elderly with hip fx die within the first year Risk Factors for Osteoporosis Age Gender Genetics Sedentary life style Smoking Alcohol Low calcium / Vitamin D High protein intake (NIH Consensus Statement, 2) The Controversy High protein diets are a risk for osteoporosis Especially if source is animal protein and calcium intake is low

The Evidence Purified sources of protein increased urinary calcium loss (Linkswiler et al 197s) common sources of protein did not (Spencer et al,1978; Hunt, et al 1995) Phosphorus Effect? Observational Evidence Feskanich et al (1996): Animal protein associated with increased risk of forearm fractures Munger et al (1999): Animal protein associated with reduced incidence of hip fractures Hannan et al (2): Lower total and animal protein intake associated with increased bone loss Wengreen et al (24): Increased total protein intake associated with reduced hip fx in younger elderly Bone Composition: 7% mineral (4% calcium; 99% body Ca) 25% organic (>9% protein) 5-8% water Function Structure, locomotion, hematopoesis, growth factors Acid-base balance Renal Mechanisms for Acid-Base Balance Animal Protein Intake Sulfur Amino Acid Intake Research Questions Net Endogenous Acid Osteoclastic activity Renal Net Acid Excretion Glomerular Filtration Rate Renal Reabsorption of Calcium Does increased animal protein intake decrease calcium retention? Does replacing animal protein with plant protein improve calcium retention? Do dietary calcium and protein interact in a dose-dependent manner?

Design Considerations Purified vs. Common protein Equalized P vs. Variable P Urinary Ca vs. Ca retention Balance vs. Tracer methodology (absorption vs. retention) Short-term vs. Long-term feeding Few subjects vs. Statistical power Protocol Volunteers: - healthy post-menopausal women Design: - randomized crossover Diets: - weighed diets; 7-8 wk Calcium Retention: - wk 3-4: labeled entire 2-d menu with 47 Ca (constant specific activity) - whole body scintillation counting for 4 wk Diet Composition 1 Low Meat High Meat Meat (beef, chicken, turkey, pork), g 65 112 Protein, % of energy 12 2 g/kg bw.94 1.62 Fat, % of energy 3 31 Carbohydrate, % of energy 58 49 Calcium, mg 7 (594.2 ± 19.3) 698 (574.1 ± 18.4) Phosphorus, mg 1214 (1218.2 ± 87.) 1659 (1616.4 ± 14.1) Potassium, mg (283 ± 339) (2184 ± 29) 1 Per 22 Kcal. Data in parentheses are analyzed values (mean ± SD) Calcium Retention in Postmenopausal Women Consuming High and Low Meat Diets for 8 wk each (Mean ±SEM) Calcium Retention (%) 1 9 8 7 6 5 4 3 2 1 Observed 5 1 15 2 25 3 Time (days) High Meat Low Meat Calcium Retention (% ) 1 9 8 7 6 5 4 3 2 1 Modeled 5 1 15 2 25 3 Time (days) High Meat Low Meat Diet: P =.9 Roughead, Johnson, Lykken & Hunt, (23) J. Nutr. 133:12 45 4 35 3 25 2 15 1 5 Titratable Acidity Diet: P <.1 Diet*Time: P <.5 Ammonium Diet: P <.1 3 5 8 3 5 8 week 3 5 8 3 5 8 High Meat Low Meat High Meat Low Meat Roughead, Johnson, Lykken & Hunt, (23) J. Nutr. 133:12

Diet did not Affect Biomarkers of Bone Metabolism Blood: Tartarate-resistant acid phosphatase, Bonespecific alkaline phosphatase, Osteocalcin, 25-OH Vitamin D Urine: Calcium, N-telopeptide Net Endogenous Acid Protein Intake Sulfur Amino Acid Intake Osteoclastic activity Renal Net Acid Excretion Glomerular Filtration Rate Renal Reabsorption of Calcium Roughead, Johnson, Lykken & Hunt, (23) J. Nutr. 133:12 Plant vs. Animal Proteins? Objectives In healthy postmenopausal women, determine if daily substitution of 25 g of soy protein for meat protein 1) improves calcium retention 2) affects biomarkers of bone metabolism Diet Composition 1 CONTROL SOY Meat (beef, chicken, pork), g 17 55 Soy Protein Isolate, g* --- 3 Protein, % of energy 15 16 g/kg bw 1.32 1.33 Fat, % of energy 3 3 Carbohydrate, % of energy 55 55 Calcium, mg 69 (653 ± 4) 746 (74 ±27) Phosphorus, mg 155 (1563 ± 27) 1533 (161 ± 178) Phytate, mg 1673 215 1 Per 22 Kcal. Data in parentheses are analyzed values (mean ± SD) *Soy protein: 2.28 mg aglycone/g protein Calcium Retention (% of dose) 1 9 8 7 6 5 4 3 2 1 Consuming 25 g of Soy Protein Substituted for Meat Protein for 8 wk did not Affect Calcium Retention in Postmenopausal Women (Mean + SE) 1 Observed 9 Modeled Modeled 8 7 CONTROL SOY 6 5 4 Diet: NS (n=12) 3 2 1 5 1 15 2 25 3 5 1 15 2 25 3 Time (day) Time (day) Roughead et al (25) J. Clin. End. Met., in press

1 Initial Absorption 6 5 Ammonium (Diet: NS) Titratable Acidity (Diet: P =.3) 4 1 Soy: 27. + 7. Control: 26.1 + 7. NS 3 2 1 1 5 1 15 2 25 3 Time, days 3 5 7 3 5 7 3 5 7 3 5 7 week CONTROL SOY CONTROL SOY Roughead et al (25) J. Clin. End. Met., in press The bottom line No evidence of adverse effects from animal protein intake on calcium metabolism even when calcium intake is low What about when Ca intake is high? n=27 Controlled Diets (HC:15 mg/d or LC: 6 mg/d) 47 Ca Whole body Counting (28 days) 1 2 3 5 7 A high protein diet increased total renal acid excretion regardless of calcium intake 12 HCHP 1 8 6 HCLP LCHP LCLP Blood Urine Urine Urine Blood Urine Urine Blood Urine Note: Participant consumed either a Low or High Protein diet for 7 weeks; after a 2-wk washout, consumed the alternate diet for 7 weeks. 4 2 Ca: NS Pro: P <.1 Ca x Pro: NS wk 1 wk 2 wk 3 wk 5 wk 7

25 2 15 1 5 Both dietary calcium and protein independently influenced urinary calcium loss wk 1 wk 2 wk 3 wk 5 wk 7 HCHP HCLP LCHP LCLP Ca: P <.1 Pro: P <.1 Ca x Pro: NS 1 9 8 7 6 5 4 3 2 1 A high protein intake enhanced calcium retention from a low calcium diet High Ca, High Protein High Ca, Low Protein Low Ca, High Protein Low Ca, Low Protein Calcium: P <.1 Protein: P<.2 Ca x Pro: P <.5 5 1 15 2 25 3 Time (days) A high protein intake enhanced calcium bioavailability from a low calcium diet 3 25 2 Calcium: P <.1 Protein: P<.2 Ca x Pro: P <.5 15. 1. A high protein intake reduced bone resorption as indicated by urinary Dpd excretion 15 1 5 5. Ca: NS Pro: P <.1 Ca x Pro: NS HCHP HCLP LCHP LCLP. HCHP HCLP LCHP LCLP ng/ml 2 15 1 5 A high protein diet increased serum IGF-1 by ~27%, regardless of calcium intake HCHP HCLP LCHP LCLP Ca: NS Pro: P<.1 Ca x P: NS In ovx rats, the efficiency of calcium retention was highest when protein intake was high (especially from animal protein) 1 9 8 7 6 5 4 3 2 1 1 2 3 7 8 9 1 13 14 Day Source: P<.3 Quantity: P <.1 Interaction: NS 2% Meat 1% Meat 2% Soy 1% Soy

12 Serum IGF-1 was higher in the rats fed the high protein diets regardless of protein source Milk Constituents 1 8 6 4 2 Source: NS Quantity: P =.2 Interaction: NS P Mg Lactose Ca Vitamin D Protein??? K Meat Soy 1% 2% 1% 2% Dairy Protein and Bone? Milk and Bone Health IsolatedCasein and Lactalbumin have been shown to be calciuric IsolatedWhey Protein may reduce bone resorption Milk supplementation, 12 mo: increased spine and total BMD in young girls (Chan, 1995) Milk supplementation, 18 mo: increased serum IGF-I and lumbar BMD and total BMC in young girls (Cadogan, 1997) Milk supplementation, 3 y: prevented vertebral bone loss in pre-meneopausal women (Baran, 199) High Ca milk supplementation 24 mo: decreased bone loss in femoral neck, lumbar spine and total hip in postmenopausal women (Chee, 23) Milk Supplementation 12 wk: increased serum IGF-I and decreased urinary N-telopeptides in older men and women (Heaney, 1999) Protein Intake Sulfur Amino Acid Intake Serum IGF-1? Future Directions X,? X X Acid Excretion Increased urine Ca loss Calcium Retention Carefully controlled feeding trials are needed to assess: Effects of dairy proteins provided as food (milk, cheese, yogurt, etc) on calcium retention and bone health Effects of dairy proteins per se on calcium retention and bone health?

As we analyze a thing into its parts and properties, we tend to magnify these, to exaggerate their apparent independence and to hide from ourselves the essential integrity and individuality for the composite whole. We may study them apart but it is a concession to our weakness and the narrow outlook of our minds. (D Arcy Thompson, 1942) Acknowledgments - Janet Hunt, PhD, RD - Bonnie Hoverson, RD - Glenn Lykken, PhD - Debbie Krause, RD - LuAnn Johnson - Brenda Hanson, RD - Carol Zito - Angela Scheett, RD - Jennifer Hanson - Emily Neilsen, RN Jackie Nelson Aldrin Lafferty Study Volunteers Primarily supported by USDA Partial support: National Cattlemen s Beef Association, North Dakota Beef Commission Solea Company (soy protein isolate)