Lab Anim Res.  2013 Jun;29(2):70-76. 10.5625/lar.2013.29.2.70.

Treatment of eggshell with casein phosphopeptide reduces the severity of ovariectomy-induced bone loss

• Kim JH
• Kim MS
• Oh HG
• Lee HY
• Park JW
• Lee BG
• Park SH
• Moon DI
• Shin EH
• Oh EK
• Erkhembaatar M
• Kim O
• Lee YR
• Chae HJ

• KMID: 1431335
• DOI: http://doi.org/10.5625/lar.2013.29.2.70

Abstract

It has been generally accepted that calcium intake prevents bone loss, and frequent fracture resulted from osteoporosis. However, it is still elusive as to how effective sole calcium intake is in preventing or attenuating the severity of osteoporosis. Here, we demonstrate the effects of eggshell-casein phosphopeptide (ES-CPP), and compared these effects those of calcium supplement, for restoring ovariectomy-mediated bone loss. CPP, synthesized from the hydrolysis of casein (0.5%) using trypsin, was added to the grinded ES and was then administered to the ovariectomized (OVX) rat at 100 mg/kg for 4 weeks. Urine and feces from each group were collected each day, and were used to calculate the apparent calcium absorption rate in a day. After 4 weeks incubation, blood and femoral bones were isolated for the analysis of parameters representing osteoporosis. The apparent calcium absorption rate was significantly increased in the ES-CPP treated groups, in comparison to both the OVX and the commercial calcium supplement (CCS) treated group. Notably, treatment with ES-CPP markedly enhanced the calcium content in femoral bone and the relative weight of femoral bone to body weight, though calcium content in serum was barely changed by treatment with ES-CPP. Parameters of osteoporosis, such as osteocalcin in serum and bone mineral density, were rescued by treatment with ES-CPP, compared to treatment with commercial calcium supplement. This finding strongly suggests the possible use of ES-CPP in preventing or attenuating the severity of postmenopausal osteoporosis.

Keyword

Osteoporosis; casein phosphopeptide; calcium; inorganic phosphorous; bone mineral density

MeSH Terms

Absorption
Animals
Body Weight
Bone Density
Calcium
Caseins
Feces
Female
Humans
Hydrolysis
Osteocalcin
Osteoporosis
Osteoporosis, Postmenopausal
Piperazines
Rats
Trypsin
Calcium
Caseins
Osteocalcin
Piperazines
Trypsin

Figure

• 

  Figure 1 Administration of ES-CPP markedly enhances the apparent Ca2+ absorption rate. Effects of ES-CPP on (A) the general symptoms of OVX-mediated osteoporosis and (B) the apparent Ca2+ absorption rate. The intake of food and water were calculated by measuring the weight of ingested food and water in a day. The weight of body, liver, and kidney were each measured after 7 weeks of administrating ES-CPP and CCS. The apparent Ca2+ absorption rate was calculated from the content of Ca2+ in urine and feces, as described in "Materials and Methods". Each column represents the proportion of absorbed Ca2+ in the body, from the administration of ES-CPP and CSS. Data are means±SE. a,b,cValues in the row with different superscript letters are significantly different, P<0.05. ES; eggshell, CPP; casein phosphopeptide, CCS; commercial calcium supplement.

• 

  Figure 2 Loss of calcium from femoral bone is restored by ES-CPP administration, regardless of calcium content in serum. Effects of ES-CPP on calcium content in (A) serum and (B) femoral bone. Calcium in serum and femoral bone was measured from isolated blood and femoral bone from OVX rat, which was administrated with ES-CPP and CCS for 7 weeks after ovariectomy. Data are mean±SE. a,b,cValues in the row with different superscript letters are significantly different, P<0.05. ES; eggshell, CPP; casein phosphopeptide, CCS; commercial calcium supplement.

• 

  Figure 3 Administration of ES-CPP attenuates the severity of osteoporosis. Effects of ES-CPP on (A) the weight of femoral bone, (B) bone mineral density, and (C) osteocalcin in serum. Isolated femoral bone segments from OVX rat were weighed and then immediately used to analyze bone mineral density using Micro-CT. Osteocalcin was analyzed from collected blood. Data are means±SE. a,b,c,dValues in the row with different superscript letters are significantly different, P<0.05. ES; eggshell, CPP; casein phosphopeptide, CCS; commercial calcium supplement.

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