Improvement by nature

Collagen peptides and Joint & Bone HealthRousselot® studies and literature

BonesBones, which makes up the skeleton, are

composed of cells embedded in hard intercel-

lular material (the matrix) made of mineralized

substances and collagen fibers. Like all body

tissues, bones are in a continuous state of flux.

The mechanical integrity of the skeleton and the

maintenance of appropriate mineral levels depend

on a dynamic process called bone remodeling or

bone turnover.

Skeletal mass increases progressively during

growth. The age at which bone loss starts is

uncertain but, it is generally believed to be during

the thirties in both sexes. Approximately 15%

of bone in healthy adults is replaced by bone

turnover each year. Irregular rates of bone resorp-

tion and formation – leading to more bone loss

than formation – are a hallmark to osteopenia.

According to the World Health Organiza-

tion (WHO), osteopenia is used to describe

individuals who have a low bone mass and some

increased risk of fracture. Their bone mass is not

so low that they are deemed to have osteoporosis.

The bone mass is measured on a point scale,

called a “T” score. The range defining osteopenia

or low bone mass is quite large, ranging from

-1 to -2.5 T-scores (that is, bone density levels

that are 1 to 2.5 standard deviations below the

average for young adults who have achieved

normal peak bone mass), which translates into

bone mass that is 10–30 percent below this

average level.

The main goal for screening and treating for

osteopenia is to maintain bone health and prevent

fractures. It has been demonstrated that an early

diagnosis and treatment of osteopenia reduces

fractures rates and improves life quality (3). In a

population-based random study (616 women

aged 60 - 94 years followed for a median 5.6

years) conducted by Pasco and published in

2006 in Osteoporosis journal (13), it has been

determinated that 73.1% of fractures occurred in

women without osteoporosis (56.5% in women

with osteopenia and 16.6% in women with normal

Bone Mineral Density).

Because life expectancy is increasing around the

world, the number of elderly individuals is rising

in every region. According to WHO, the number

of individuals aged 65 years or over will increase to

an estimated 1555 million by the year 2050. The

influence of such changes on the number and

regional distribution of hip fractures will be dramatic

as the population burden of fractures originates in

men and women with osteopenia.

As the population ages, the drawbacks of aging are becoming more

apparent. Osteoarthritis and osteopenia, two of the major health concerns,

are among the leading causes of pain and disability.

It is now well known that collagen peptides can help to maintain bone and

joint health to prevent osteopenia and osteoarthritis.

Hip

fra

ctu

res

(103 )

Joint & Bone HealthAging of the world’s population sees growth in bone and joint health problems.

2500

2000

1500

1000

500

0

Men WomenNorth

America

Men WomenEurope

Men WomenRussian

Federation

Men WomenEastern

Mediterranean

Men WomenAsia

Men WomenAfrica

1990

2025

2050

Number of hip fractures in 1990 and those predicted in different regions of the world for 2025 and 2050

+1,0 –– 0,0 ––

-1,0 –– -2,0 –– -2,5 –– -3,0 –– -4,0 ––

Normal bone density

Low bone mass

Presence of osteoporosis

Bone mineral T-score measure

Joints Cartilage consists of a single cell type, chondro-

cytes embedded in extracellular matrix made up

of two major components: type II collagen which

imparts tensile strength to the tissue and aggrecan

that provides the ability of cartilage to resist

compressive force. Orchestrated synthesis and

turnover ensures and maintains the biochemical

characteristic of the cartilage.

During osteoarthritis, the most common form of

arthritis, this regulation is disrupted by the expres-

sion of pro-inflammatory molecules that provides

the stimulus for the synthesis of matrix-degrading

enzymes. These enzymes degrade aggrecan and

collagen, resulting in the loss of cartilage and

function.

This chronic degeneration of the joints is already

considered by the World Health Organization as

one of the most disabling diseases in developed

countries. For those who are affected, the

consequences are important because 80% have

limitations in movement, and 25% can not

perform their major daily activities of life.

All joints can be affected, although it occurs more

often on knee and hip joints and in the vertebral

column. In advanced stages it is extremely painful.

Osteoarthritis is usually due to the

aging process, but osteoarthritis

due to external factors has

also been reported, for

example in people whose

activities involve joint

stress, such as sports

activities or even at work.

Severe overweight can

also cause osteoarthritis.

Prevalence of osteoarthritis

Mill

ion

s o

f p

eop

le

160

140

120

100

80

60

40

20

0India China

2003

2008

2013

US JapanBrazil Germany France

Arthritis in the world

According to worldwide estimation, arthritis

affects 9.6% of men and 18% of women aged

over 60 years.

Moreover, according to Datamonitor (2009), the

arthritis prevalence has increased all over the

world since 2003, such as in India (+2.3% per

year) or Brazil (+2.1% per year), and will continue

to grow, possibly even reaching 2.5% per year in

India and 2.3% per year in Brazil.

Peptan®

Collagen peptides for bone health

Collagen, which represents 90% of organic

bone mass, has been identified as effective in the

treatment of osteopenia. Several studies show

that a daily intake of 10g of collagen peptides for

4 to 24 weeks increases bone mass density (1, 2).

Researchers have produced several keys for

understanding how collagen peptides work.

They have also shown that the extra-cellular

matrix in which cells grow is decisive in their

differentiation. Studies have demonstrated that

when collagen peptides metabolites are present in

this matrix, osteoblasts, the cells responsible for

bone formation, are preferentially stimulated,

instead of osteoclasts, the cells involved in bone

resorption, this triggers bone formation (4, 5, 6).

Studies conducted by Rousselot with Peptan® at

the Physiology and Ingestive Behavior Laboratory,

INRA-AgroParis Tech (Paris, France) and presented

below, confirm these scientific findings and

provide new information about the effect of

Peptan® on bone metabolism (7).

A culture of osteoblasts (star-shape cells) and osteoclasts (round black cells) with Peptan®.

Peptan® induces the differentiation of cells

into osteoblasts

In mixed cultures of osteoblasts and osteoclasts,

the levels of alkaline phosphatase (ALP), a marker

of bone formation, are increased when Peptan®

is present in the media. Since the number of cells

does not increase with Peptan® compared to the

control group, this means that Peptan® induces

the differentiation of cells into osteoblasts, rather

than osteoclasts.

Peptan® reduces bone resorption

Moreover, when the culture is performed on a system which allows measurement of

the osteoclast-mediated bone resorption, we can see that when Peptan® is present

the resorption area is reduced compared to the control.

Measures of Alkaline Phosphatase

mg

/mL/

min

/∆D

C

2,5

2

1,5

1

0,5

0BSA Peptan® B

1 mg/mL

0,5 mg/mL

Peptan® P Peptan® F

Measures of alkaline phosphatase (ALP) activity in bone cell culture with Peptan® B (bovine), Peptan® P (porcine) and Peptan® F (fish) for 14 days, compared to BSA (standard proteins).

*

** *

*

*

A B

Rousselot® in vitro results

Cell culture with BSA standard protein (A) or with Peptan® (B): osteoclasts activity is measured by the digestion of a calcium phosphate film (white spot).

Peptan® induces differentiation into osteoblast, at the same time reducing

the differentiation into osteoclast.

Rousselot® in vivo results

BMD increase

(g/c

m2 )

0,025

0,02

0,015

0,01

0,005

0Control

a

b

a

Ovx Ovx+Peptan®

Measure of the cortical area

(mm

2 )

1,25

1,2

1,15

1,1Control

ab

b

a

Ovx Ovx+Peptan®

Measure of CTX concentration

(µg

/mL)

15

10

5

0Control

b

a

b

Ovx Ovx+Peptan

Measure of ultimate strengthN

31

30

29

28

27

26

25Control

abb

a

Ovx Ovx+Peptan®

Increase of Bone Mineral Density (BMD) of the mice, in the control group (Control), the ovariectomized group fed without Peptan® (Ovx), and the ovariectomized group fed with Peptan® (Ovx + Peptan) after 12 weeks. Groups with different letters indicate significant difference (p<0.05).

The in vivo part of this study confirms but

also reveals some completely new findings.

In this part, ovariectomized mice were used to

simulate low bone mass density: osteopenia.

This surgery actually slows down the bone

mineral density increase during the growth of

the concerned mice. The mice were fed with or

without Peptan® for 12 weeks and various data

was collected. The Peptan® group had a 4g daily

diet containing 2.5% (0.1g) of Peptan®.

First, in ovariectomized mice fed with Peptan® for

12 weeks, measures show restoration of the bone

mineral density value close to the level of the

control group (which was not ovariectomized).

Moreover, growth of the cortical zone (external

area of the bone) of the femur, and a significant

increase in bone size, was measured in ovariec-

tomized mice fed with Peptan®, compared to

ovariectomized mice not fed with Peptan®.

Due to this restoration, the ultimate strength of

the bones (strength required to lead to a break)

was significantly greater for ovariectomized mice

fed with Peptan®.

Finally, confirming the in vitro studies, in vivo

studies have demonstrated that the bone

resorption of ovariectomized mice fed Peptan®,

measured in terms of CTX production, is similar to

the control group.

Measure of the cortical area and the ultimate strength, in the control group (Control), the ovariectomized group fed without Peptan® (Ovx), and the ovariectomized group fed with Peptan® (Ovx + Peptan).Groups with different letters indicate significant difference (p<0.05) - ab is not significantly different from a or b.

Peptan® increases bone size and solidity

Measure of the Carboxy-Terminal collagen (CTX) peptides concentration, peptides fragments generated by collagen degradation, an important biochemical marker of bone resorption, in the control group (Control), the ovariectomized group fed without Peptan® (Ovx), and the ovariectomized group fed with Peptan® (Ovx + Peptan). Groups with different letters indicate significant difference (p<0.05).

Peptan® has been shown to be effective on

bone metabolism by inducing differentiation

and maturation of osteoblast and stimulat-

ing their activity. Bone turnover is thus

modulated, preventing bone resorption

during the natural phenomenon of bone loss

(osteopenia) and increasing bone solidity.

Peptan® restores bone mineral density

Peptan® decreases bone resorption

a placebo. At the end of the study, an assessment

of participants by physicians showed statistically

significant changes with the dietary supplement

collagen peptides: joint pain at rest, when walking,

standing or carrying objects was reduced (10).

Peptan® A bioactive ingredient that reduces joint pain

Collagen peptides offer proven efficacy

The beneficial effects of collagen peptides have

been reported in numerous research findings.

The clinical study conducted by Moskowitz (8),

performed on various populations for a period

ranging from 30 to 90 days, demonstrated a

positive effect on pain reduction with a dose of

10g of collagen peptides per day. The effect was

even more pronounced in patients suffering more

severe symptoms.

Another more recent study assessed the efficiency

of collagen peptides in a randomized, double

blind, controlled multicentre trial, in which 250

subjects with primary osteoarthritis

of the knee were given 10g of

collagen peptides daily. The results

showed a significant improvement in

knee joint comfort, and again subjects

with the greatest joint deterioration

benefited most (9).

The effectiveness of collagen peptides has been

also proven in athletes with activity-related joint

pain. In a 24-week study conducted in 2008, 147

subjects who compete as part of a varsity team or

club sport were recruited and randomly assigned

to two groups: one group received a liquid with

10g of collagen peptides, and the other received

Type ll collagen secretion by chondrocytes in culture

Typ

e ll

colla

gen

(μ

g/1

06 ch

on

dro

cyte

s)

Culture time (days)

1,8

1,6

1,4

1,2

1

0,8

0,6

0,4

0,2

06 8 11

Basal medium (control)

Medium containingcollagen peptides

Collagen peptides in cartilage

Rad

ioac

tivi

ty (

dm

p/1

00 m

g c

arti

lag

e)

Time (hours)

Control14C labeled collagen peptides

300

250

200

150

100

50

0

0 12 24 36 48 60 72 84 96

Collagen peptides are accumulated in cartilageSome explanations of this positive effect were revealed in the study by Oesser (11). He showed that more

than 95% of 14C labeled collagen peptides taken orally is absorbed. However, he also showed that an

important part is accumulated in cartilage of mice as early as 12 hours after ingestion. The radioactivity

level was more than twice as high in cartilage following 14C labeled collagen peptide administration

compared with the control group (14C labeled proline + collagen peptides).

Time course of radioactivity in cartilage of mice subsequent to absorption of 14C labeled collagen peptides and 14C labeled proline in the control group. The mice received 10mg collagen peptides/g body weight. *p<0.05 between collagen peptides and control (11).

Collagen peptides induce the production of type II collagen by chondrocytes

In another study (12), the same author explains that the chondrocytes (cells of the cartilage) are sensitive

to the products present in the extracellular matrix around them, and that the treatment of chondrocytes

with collagen peptides in the medium leads to a dose-dependent production of collagen by those cells.

Time course of type II collagen secretion into supernatant of bovine chondrocytes cultured in basal medium or in medium supplemented with 0.5 mg/mL of collagen peptides. After 11 days of culture, type II collagen secretion was almost 2.5-fold higher in collagen peptides cultures. *p<0.01 between collagen peptides and control cultures (12).

**

**

*

*

*

The action mechanism of collagen peptides

Regarding bones, Peptan® has been shown

to:

• Restore bone mineral density.

• Increase the bones size, making them less

brittle.

• Stimulate osteoblast activity in spite of

osteoclasts activity; the growth of the bone is

stimulated.

Regarding joints, collagen peptides have been

shown to:

• Significantly reduce joint pain, and subjects with

the greater deterioration benefit most.

• Orally taken collagen peptides are accumulated

in cartilage as soon as 12 hours after the inges-

tion and the cells of the joint, the chondrocytes,

are responding to these extern collagen by

producing a dose-dependant production of

intern collagen.

Conclusion

Peptan®

a completely characterized and scientifically objectivised bioactive ingredient that helps to:

•Prevent osteopenia

• Maintain bone health

• Prevent loss of bone solidity

•Prevent osteoarthritis

• Reduce joint pain

• Prevent joint matrix degeneration

These different studies demonstrate the benefits of collagen peptides on joint and bone health.

The results indicate that collagen peptides could be useful in the prevention and treatment of osteopenia

and osteoarthritis.

References

1. Nomura, Y., Oohashi, K., Watanabe, M. and Kasugai, S. 2005. Increase in bone mineral density through oral administration of shark gelatine to ovariectomized rats. Nutrition, 21: 1120-1126.

2. Wu, J., Fujioka, M., Sugimoto, K., Mu, G. and Ishimi, Y. 2004. Assessment of effectiveness of oral administration of collagen peptide on bone metabolism in growing and mature rats. Journal of bone and mineral metabolism, 22: 547-553.

3. Karaguzel, G., Holick, M. 2010. Diagnosis and treatment of osteopenia. Rev Endocr Metab Disord, 11: 237-251.4. Mizuno, M. and Kuboki, Y. 2001. Osteoblast-related gene expression of bone marrow cells during the osteoblastic differentiation induced by type I collagen. Journal of biochemistry,

129: 133-138.5. Andrianarivo, A.G., Robinson, J.A., Mann, K.G. and Tracy R.P. 1992. Growth on type I collagen promotes expression of the osteoblastic phenotype in human osteosarcoma MG-63 cells.

Journal of cellular physiology, 153: 256-265.6. Lynch, M.P., Stein, J.L., Stein, G.S. and Lian, J.B. 1995. The influence of type I collagen on the development and maintenance of the osteoblast phenotype in primary and passaged rat

calvarial osteoblasts: modification of expression of genes supporting cell growth, adhesion, and extracellular matrix mineralization. Experimental cell research, 216: 35-45.7. Guillerminet, F., Beaupied, H., Fabien-Soulé, V., Tomé, D., Benhamou, C-L., Blachier, F., Roux, C. and Blais, A. 2010. collagen peptides improves bone metabolism and biomechanical

parameters in ovariectomized mice: an in vitro and in vivo study. Bone. 8. Moskowitz, R. 2000. Role of collagen hydrolysate in bone and joint disease. Seminars in arthritis and rheumatism, 30 (2): 87-99. 9. Ruiz-Benito, P., Camacho-Zambrano, M.M., Carrillo-Arcentales, J.N., Mestanza-Peralta, M.A., Vallejo-Flores, C.A., Vargas-Lopez, S.V., Villacis-Tamayo, R.A. and Zurita-Gavilanes, L.A.

2009. A randomized controlled trial on the efficacy and safety of a food ingredient, collagen hydrolysate, for improving joint comfort. International journal of food sciences and nutrition, 12:1-15.

10. Clark, K.L., Sebastianelli, W., Flechsenhar, K.R., Aukermann, D.F., Meza, F., Millard, R.L., Deitch, J.R., Sherbondy, P.S. and Albert, A.. 2008. 24-Week study on the use of collagen hydrolysate as a dietary supplement in athletes with activity-related joint pain. Current medical research and opinion, 24 (5): 1485-1496.

11. Oesser, S., Adam, M., Babel, W. and Seifert, J. 1999. Oral administration of 14C labelled gelatine hydrolysate leads to an accumulation of radioactivity in cartilage of mice (C57/BL). Journal of nutrition, 129: 1891-1895.

12. Oesser, S. and Seifert, J. 2003. Stimulation of type II collagen biosynthesis and secretion in bovine chondrocytes cultured with degraded collagen. Cell tissue research, 311: 393-399. 13. Pasco, J.A, Seeman, E., Henry M.J, Merriman, E.N, Nicholson, G.C, Kotowicz, M.A. 2006. The population burden of fractures originates in women with osteopenia, not osteoporosis.

Osteoporosis Int, 17: 1404-1409.

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Rousselot®

Rousselot is the leading manufacturer of gelatine

and collagen peptides to the food, pharma ceutical

and technical industries. With a staff of 2,400

people, the company benefits from a global sales

and production network of 13 plants and 10 sales

offices located in Europe, North America, South

America and Asia. Rousselot is part of VION N.V.,

an international food company with production

and sales facilities on all continents.

VION

With two international divisions, Food and

Ingredients, the company is active in the field of

high quality foodstuffs and health products for

humans and animals. Rousselot is part of the

Ingredients division. VION has annual sales of

EUR 9.0 billion and provides employment for

27,000 people worldwide. VION‘s head office is

in Eindhoven, The Netherlands.

www.vionfood.com

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