Skip to main content
Top
Published in: Osteoporosis International 2/2015

01-02-2015 | Original Article

Kefir improves bone mass and microarchitecture in an ovariectomized rat model of postmenopausal osteoporosis

Authors: H.-L. Chen, Y.-T. Tung, C.-H. Chuang, M.-Y. Tu, T.-C. Tsai, S.-Y. Chang, C.-M. Chen

Published in: Osteoporosis International | Issue 2/2015

Login to get access

Abstract

Summary

Kefir treatment in ovariectomized (OVX) rats could significantly decrease the levels of bone turnover markers and prevent OVX-induced bone loss, deterioration of trabecular microarchitecture, and biomechanical dysfunction that may be due to increase intracellular calcium uptake through the TRPV6 calcium channel.

Introduction

Osteoporosis is a disease characterized by low bone mass and structural deterioration of bone tissue, leading to an increased fracture risk. The incidence of osteoporosis increases with age and occurs most frequently in postmenopausal women due to estrogen deficiency, as the balance between bone resorption and bone formation shifts towards increased levels of bone resorption. Among various methods of prevention and treatment for osteoporosis, an increase in calcium intake is the most commonly recommended preventive measure. Kefir is a fermented milk product made with kefir grains that degrade milk proteins into various peptides with health-promoting effects, including immunomodulating-, antithrombotic-, antimicrobial-, and calcium-absorption-enhancing bioactivities.

Methods

The aim of this study is to investigate the effect of kefir on osteoporosis prophylaxis in an ovariectomized rat model. A total of 56 16-week-old female Sprague-Dawley (SD) rats were divided into 7 experimental groups: sham (normal), OVX/Mock, OVX/1X kefir (164 mg/kg BW/day), OVX/2X kefir (328 mg/kg BW/day), OVX/4X kefir (656 mg/kg BW/day), OVX/ALN (2.5 mg/kg BW/day), and OVX/REBONE (800 mg/kg BW/day). After 12-week treatment with kefir, the bone physiology in the OVX rat model was investigated. Accordingly, the aim of this study was to investigate the possible transport mechanism involved in calcium absorption using the Caco-2 human cell line.

Results

A 12-week treatment with kefir on the OVX-induced osteoporosis model reduced the levels of C-terminal telopeptides of type I collagen (CTx), bone turnover markers, and trabecular separation (Tb. Sp.). Additionally, treatment with kefir increased trabecular bone mineral density (BMD), bone volume (BV/TV), trabecular thickness (Tb. Th), trabecular number (Tb. N), and the biomechanical properties (hardness and modulus) of the distal femur with a dose-dependent efficacy. In addition, in in vitro assay, we found that kefir increased intracellular calcium uptake in Caco-2 cell through TRPV6 calcium channels and not through L-type voltage-operated calcium channels.

Conclusion

The protective effect of kefir in the OVX rat model may occur through increasing intracellular calcium uptake through the TRPV6 calcium channel.
Appendix
Available only for authorised users
Literature
1.
go back to reference Renno AC, de Moura FM, Dos SN, Tirico RP, Bossini PS, Parizotto NA (2006) Effects of 830-nm laser, used in two doses, on biomechanical properties of osteopenic rat femora. Photomed Laser Surg 24:202–206PubMedCrossRef Renno AC, de Moura FM, Dos SN, Tirico RP, Bossini PS, Parizotto NA (2006) Effects of 830-nm laser, used in two doses, on biomechanical properties of osteopenic rat femora. Photomed Laser Surg 24:202–206PubMedCrossRef
2.
go back to reference Genant HK, Delmas PD, Chen P, Jiang Y, Eriksen EF, Dalsky GP, Marcus R, San MJ (2007) Severity of vertebral fracture reflects deterioration of bone microarchitecture. Osteoporos Int 18:69–76PubMedCrossRef Genant HK, Delmas PD, Chen P, Jiang Y, Eriksen EF, Dalsky GP, Marcus R, San MJ (2007) Severity of vertebral fracture reflects deterioration of bone microarchitecture. Osteoporos Int 18:69–76PubMedCrossRef
3.
go back to reference Chen H, Wu M, Kubo KY (2012) Combined treatment with a traditional Chinese medicine, Hachimi-jio-gan (Ba-Wei-Di-Huang-Wan) and alendronate improves bone microstructure in ovariectomized rats. J Ethnopharmacol 142:80–85PubMedCrossRef Chen H, Wu M, Kubo KY (2012) Combined treatment with a traditional Chinese medicine, Hachimi-jio-gan (Ba-Wei-Di-Huang-Wan) and alendronate improves bone microstructure in ovariectomized rats. J Ethnopharmacol 142:80–85PubMedCrossRef
4.
go back to reference Naot D, Grey A, Reid IR, Cornish J (2005) Lactoferrin—a novel bone growth factor. Clin Med Res 5:93–101CrossRef Naot D, Grey A, Reid IR, Cornish J (2005) Lactoferrin—a novel bone growth factor. Clin Med Res 5:93–101CrossRef
5.
go back to reference Shen Y, Li YQ, Li SP, Ma L, Ding LJ, Ji H (2010) Alleviation of ovariectomy-induced osteoporosis in rats by Panax notoginseng saponins. J Nat Med 64(3):336–345PubMedCrossRef Shen Y, Li YQ, Li SP, Ma L, Ding LJ, Ji H (2010) Alleviation of ovariectomy-induced osteoporosis in rats by Panax notoginseng saponins. J Nat Med 64(3):336–345PubMedCrossRef
6.
7.
go back to reference Cho DC, Kim KT, Jeon Y, Sung JK (2012) A synergistic bone sparing effect of curcumin and alendronate in ovariectomized rat. Acta Neurochir 154(12):2215–2223PubMedCrossRef Cho DC, Kim KT, Jeon Y, Sung JK (2012) A synergistic bone sparing effect of curcumin and alendronate in ovariectomized rat. Acta Neurochir 154(12):2215–2223PubMedCrossRef
8.
go back to reference Zhao X, Wu ZX, Zhang Y, Yan YB, He Q, Cao PC, Lei W (2011) Anti-osteoporosis activity of Cibotium barometz extract on ovariectomy-induced bone loss in rats. J Ethnopharmacol 137(3):1083–1088PubMedCrossRef Zhao X, Wu ZX, Zhang Y, Yan YB, He Q, Cao PC, Lei W (2011) Anti-osteoporosis activity of Cibotium barometz extract on ovariectomy-induced bone loss in rats. J Ethnopharmacol 137(3):1083–1088PubMedCrossRef
9.
go back to reference Lopitz-Otsoa F, Rementeria A, Elguezabal N, Garaizar J (2006) Kefir: a symbiotic yeasts-bacteria community with alleged healthy capabilities. Rev Iberoam Micol 23:67–74PubMedCrossRef Lopitz-Otsoa F, Rementeria A, Elguezabal N, Garaizar J (2006) Kefir: a symbiotic yeasts-bacteria community with alleged healthy capabilities. Rev Iberoam Micol 23:67–74PubMedCrossRef
10.
go back to reference Guzel-Seydim ZB, Kok-Tas T, Greene AK, Seydim AC (2011) Review: functional properties of kefir. Crit Rev Food Sci Nutr 51:261–268PubMedCrossRef Guzel-Seydim ZB, Kok-Tas T, Greene AK, Seydim AC (2011) Review: functional properties of kefir. Crit Rev Food Sci Nutr 51:261–268PubMedCrossRef
11.
go back to reference de de Moreno AL, Matar C, Farnworth E, Perdigon G (2007) Study of immune cells involved in the antitumor effect of kefir in a murine breast cancer model. J Dairy Sci 90:1920–1928CrossRef de de Moreno AL, Matar C, Farnworth E, Perdigon G (2007) Study of immune cells involved in the antitumor effect of kefir in a murine breast cancer model. J Dairy Sci 90:1920–1928CrossRef
12.
go back to reference Chen HL, Tung YT, Tsai CL, Lai CW, Tsai HC, Lin YL, Wang CH, Chen CM (2014) Kefir improves fatty liver syndrome through inhibited lipogenesis pathway in leptin-deficient ob/ob knockout mice. Int J Obesity 38:1172–1179 Chen HL, Tung YT, Tsai CL, Lai CW, Tsai HC, Lin YL, Wang CH, Chen CM (2014) Kefir improves fatty liver syndrome through inhibited lipogenesis pathway in leptin-deficient ob/ob knockout mice. Int J Obesity 38:1172–1179
13.
go back to reference Vinderola G, Perdigón G, Duarte J, Farnworth E, Matar C (2006) Effects of the oral administration of the products derived from milk fermentation by kefir microflora on immune stimulation. J Dairy Res 73:472–479PubMedCrossRef Vinderola G, Perdigón G, Duarte J, Farnworth E, Matar C (2006) Effects of the oral administration of the products derived from milk fermentation by kefir microflora on immune stimulation. J Dairy Res 73:472–479PubMedCrossRef
14.
go back to reference Wronski TJ, Lowry PL, Walsh CC, Ignaszewski LA (1985) Skeletal alterations in ovariectomized rats. Calcif Tissue Int 37:324–328PubMedCrossRef Wronski TJ, Lowry PL, Walsh CC, Ignaszewski LA (1985) Skeletal alterations in ovariectomized rats. Calcif Tissue Int 37:324–328PubMedCrossRef
15.
go back to reference Wonski TJ, Dann LM, Scott KS, Crooke LR (1989) Endocrine and pharmacological suppressors of bone turnover protect against osteopenia in ovariectomized rats. Endocrinology 25:810–816CrossRef Wonski TJ, Dann LM, Scott KS, Crooke LR (1989) Endocrine and pharmacological suppressors of bone turnover protect against osteopenia in ovariectomized rats. Endocrinology 25:810–816CrossRef
16.
go back to reference Kalu DK (1991) The ovariectomized rat model of postmenopausal bone loss. J Bone Miner 15:175–192CrossRef Kalu DK (1991) The ovariectomized rat model of postmenopausal bone loss. J Bone Miner 15:175–192CrossRef
17.
go back to reference Rocha RA, Devesa V, Vélez D (2013) In vitro study of intestinal transport of fluoride using the Caco-2 cell line. Food Chem Toxicol 55:156–163PubMedCrossRef Rocha RA, Devesa V, Vélez D (2013) In vitro study of intestinal transport of fluoride using the Caco-2 cell line. Food Chem Toxicol 55:156–163PubMedCrossRef
18.
go back to reference Hidalgo IJ, Raub TJ, Borchardt RT (1989) Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability. Gastroenterology 96:736–749PubMed Hidalgo IJ, Raub TJ, Borchardt RT (1989) Characterization of the human colon carcinoma cell line (Caco-2) as a model system for intestinal epithelial permeability. Gastroenterology 96:736–749PubMed
19.
go back to reference Maubon N, Le Vee M, Fossati L, Audry M, Le Ferrec E, Bolze S, Fardel O (2007) Analysis of drug transporter expression in human intestinal Caco-2 cells by real-time PCR. Fundam Clin Pharmacol 21:659–663PubMedCrossRef Maubon N, Le Vee M, Fossati L, Audry M, Le Ferrec E, Bolze S, Fardel O (2007) Analysis of drug transporter expression in human intestinal Caco-2 cells by real-time PCR. Fundam Clin Pharmacol 21:659–663PubMedCrossRef
20.
go back to reference Lin CW, Chen HL, Liu JR (1999) Identification and characterization of lactic acid bacteria and yeasts isolated from kefir grains in Taiwan. Aust J Dairy Technol 54:14–18 Lin CW, Chen HL, Liu JR (1999) Identification and characterization of lactic acid bacteria and yeasts isolated from kefir grains in Taiwan. Aust J Dairy Technol 54:14–18
21.
go back to reference Ko YJ, Wu JB, Ho HY, Lin WC (2012) Antiosteoporotic activity of Davallia formosana. J Ethnopharmacol 139(2):558–565PubMedCrossRef Ko YJ, Wu JB, Ho HY, Lin WC (2012) Antiosteoporotic activity of Davallia formosana. J Ethnopharmacol 139(2):558–565PubMedCrossRef
22.
go back to reference Shackelford C, Long G, Wolf J, Okerberg C, Herbert R (2002) Qualitative and quantitative analysis of nonneoplastic lesions in toxicology studies. Toxicol Pathol 30(1):93–96PubMedCrossRef Shackelford C, Long G, Wolf J, Okerberg C, Herbert R (2002) Qualitative and quantitative analysis of nonneoplastic lesions in toxicology studies. Toxicol Pathol 30(1):93–96PubMedCrossRef
23.
go back to reference Wang YT, Chang SY, Huang YC, Tsai TC, Chen CM, Lim CT (2013) Nanomechanics insights into the performance of healthy and osteoporotic bones. Nano Lett 13(11):5247–5254PubMedCrossRef Wang YT, Chang SY, Huang YC, Tsai TC, Chen CM, Lim CT (2013) Nanomechanics insights into the performance of healthy and osteoporotic bones. Nano Lett 13(11):5247–5254PubMedCrossRef
24.
go back to reference Chang YT, Chen CM, Tu MY, Chen HL, Chang SY, Tsai TC, Wang YT, Hsiao HL (2011) Effects of osteoporosis and nutrition supplements on structures and nanomechanical properties of bone tissue. J Mech Behav Biomed Mater 4:1412–1420PubMedCrossRef Chang YT, Chen CM, Tu MY, Chen HL, Chang SY, Tsai TC, Wang YT, Hsiao HL (2011) Effects of osteoporosis and nutrition supplements on structures and nanomechanical properties of bone tissue. J Mech Behav Biomed Mater 4:1412–1420PubMedCrossRef
25.
go back to reference McCormack JG, Cobbold PH (1991) Cellular calcium: a practical approach. Oxford University Press, New York, NY, The practical approach series McCormack JG, Cobbold PH (1991) Cellular calcium: a practical approach. Oxford University Press, New York, NY, The practical approach series
26.
go back to reference Hertrampf T, Schleipen B, Velders M, Laudenbach U, Fritzemeier KH, Diel P (2008) Estrogen receptor subtype-specific effects on markers of bone homeostasis. Mol Cell Endocrinol 291:104–108PubMedCrossRef Hertrampf T, Schleipen B, Velders M, Laudenbach U, Fritzemeier KH, Diel P (2008) Estrogen receptor subtype-specific effects on markers of bone homeostasis. Mol Cell Endocrinol 291:104–108PubMedCrossRef
27.
28.
go back to reference Kim T, Ha H, Shim KS, Cho WK, Ma JY (2013) The anti-osteoporotic effect of Yijung-tang in an ovariectomized rat model mediated by inhibition of osteoclast differentiation. J Ethnopharmacol 146(1):83–89PubMedCrossRef Kim T, Ha H, Shim KS, Cho WK, Ma JY (2013) The anti-osteoporotic effect of Yijung-tang in an ovariectomized rat model mediated by inhibition of osteoclast differentiation. J Ethnopharmacol 146(1):83–89PubMedCrossRef
29.
go back to reference Calvo MS, Eyre DR, Gundberg CM (1996) Molecular basis and clinical application of biological markers of bone turnover. Endocr Rev 17:33–368 Calvo MS, Eyre DR, Gundberg CM (1996) Molecular basis and clinical application of biological markers of bone turnover. Endocr Rev 17:33–368
30.
go back to reference Garnero P, Delmas PD (1998) Biochemical makers of bone turnover: applications for osteoporosis. Endocrinol Metab Clin N Am 27:303–323CrossRef Garnero P, Delmas PD (1998) Biochemical makers of bone turnover: applications for osteoporosis. Endocrinol Metab Clin N Am 27:303–323CrossRef
31.
go back to reference Rosen HN, Moses AC, Garber J, Iloputaife ID, Ross DS, Lee SL, Greenspan SL (2000) Serum CTX: a new marker of bone resorption that shows treatment effect more than other markers because of low coefficient of variability and large changes with bisphosphonate therapy. Calcif Tissue Int 66:100–103PubMedCrossRef Rosen HN, Moses AC, Garber J, Iloputaife ID, Ross DS, Lee SL, Greenspan SL (2000) Serum CTX: a new marker of bone resorption that shows treatment effect more than other markers because of low coefficient of variability and large changes with bisphosphonate therapy. Calcif Tissue Int 66:100–103PubMedCrossRef
32.
go back to reference Garnero P (1996) Increased bone turnover in late postmenopausal women is a major determinant of osteoporosis. J Bone Miner Res 11:337–349PubMedCrossRef Garnero P (1996) Increased bone turnover in late postmenopausal women is a major determinant of osteoporosis. J Bone Miner Res 11:337–349PubMedCrossRef
33.
go back to reference Melton L (1997) Relationship of bone turnover to bone density and fractures. J Bone Miner Res 12:1083–1091PubMedCrossRef Melton L (1997) Relationship of bone turnover to bone density and fractures. J Bone Miner Res 12:1083–1091PubMedCrossRef
34.
go back to reference Bouxsein ML (2003) Mechanisms of osteoporosis therapy: a bone strength perspective. Clin Cornerstone S13–S21 Bouxsein ML (2003) Mechanisms of osteoporosis therapy: a bone strength perspective. Clin Cornerstone S13–S21
35.
go back to reference Peng ZQ, Vaananen HK, Zhang HX, Tuukkanen J (1997) Long-term effects of ovariectomy on the mechanical properties and chemical composition of rat bone. Bone 20:207–212PubMedCrossRef Peng ZQ, Vaananen HK, Zhang HX, Tuukkanen J (1997) Long-term effects of ovariectomy on the mechanical properties and chemical composition of rat bone. Bone 20:207–212PubMedCrossRef
36.
go back to reference Urasopon N, Hamada Y, Cherdshewasart W, Malaivijitnond S (2008) Preventive effects of Pueraria mirifica on bone loss in ovariectomized rats. Maturitas 59:137–148PubMedCrossRef Urasopon N, Hamada Y, Cherdshewasart W, Malaivijitnond S (2008) Preventive effects of Pueraria mirifica on bone loss in ovariectomized rats. Maturitas 59:137–148PubMedCrossRef
37.
go back to reference Zhang R, Liu ZG, Li C, Hu SJ, Liu L, Wang JP, Mei QB (2009) Du-Zhong (Eucommia ulmoides Oliv.) cortex extract prevent OVX-induced osteoporosis in rats. Bone 45:553–559PubMedCrossRef Zhang R, Liu ZG, Li C, Hu SJ, Liu L, Wang JP, Mei QB (2009) Du-Zhong (Eucommia ulmoides Oliv.) cortex extract prevent OVX-induced osteoporosis in rats. Bone 45:553–559PubMedCrossRef
38.
go back to reference Snyde BD, Piazza S, Edwards WT, Hayes WC, Schaffler R (1993) Role of trabecular morphology in the etiology of age-related vertebral fractures. Calcif Tissue International 53:14–22CrossRef Snyde BD, Piazza S, Edwards WT, Hayes WC, Schaffler R (1993) Role of trabecular morphology in the etiology of age-related vertebral fractures. Calcif Tissue International 53:14–22CrossRef
39.
go back to reference Kleerekope M, Villanueva AR, Stanciu J, Rao DS, Parfitt AM (1995) The role of three-dimensional trabecular microstructure in the pathogenesis of vertebral compression fractures. Calcif Tissue Int 37:594–597CrossRef Kleerekope M, Villanueva AR, Stanciu J, Rao DS, Parfitt AM (1995) The role of three-dimensional trabecular microstructure in the pathogenesis of vertebral compression fractures. Calcif Tissue Int 37:594–597CrossRef
40.
go back to reference Laib A, Barou O, Vico L, Lafage-Proust MH, Alexandre C, Rugsegger P (2000) 3D micro-computed tomography of trabecular and cortical bone architecture with application to a rat model of immobilisation osteoporosis. Med Biol Eng Comput 38:326–332PubMedCrossRef Laib A, Barou O, Vico L, Lafage-Proust MH, Alexandre C, Rugsegger P (2000) 3D micro-computed tomography of trabecular and cortical bone architecture with application to a rat model of immobilisation osteoporosis. Med Biol Eng Comput 38:326–332PubMedCrossRef
41.
go back to reference Sran MM, Boyd SK, Cooper DM, Khan KM, Zernicke RF, Oxland TR (2007) Regional trabecular morphology assessed by micro-CT is correlated with failure of aged thoracic vertebrae under a posteroanterior load and may determine the site of fracture. Bone 40:751–757PubMedCrossRef Sran MM, Boyd SK, Cooper DM, Khan KM, Zernicke RF, Oxland TR (2007) Regional trabecular morphology assessed by micro-CT is correlated with failure of aged thoracic vertebrae under a posteroanterior load and may determine the site of fracture. Bone 40:751–757PubMedCrossRef
42.
go back to reference Yarrow JF, Conover CF, Purandare AV, Bhakta AM, Zheng N, Conrad B, Altman MK, Franz SE, Wronski TJ, Borst SE (2008) Supraphysiological testosterone enanthate administration prevents bone loss and augments bone strength in gonadectomized male and female rats. Am J Physiol Endocrinol Metab 295:E1213–E1222PubMedCrossRef Yarrow JF, Conover CF, Purandare AV, Bhakta AM, Zheng N, Conrad B, Altman MK, Franz SE, Wronski TJ, Borst SE (2008) Supraphysiological testosterone enanthate administration prevents bone loss and augments bone strength in gonadectomized male and female rats. Am J Physiol Endocrinol Metab 295:E1213–E1222PubMedCrossRef
43.
go back to reference Libouban H, Blouin S, Moreau MF, Baslé MF, Audran M, Chappard D (2008) Effects of risedronate in a rat model of osteopenia due to orchidectomy and disuse: densitometric, histomorphometric and microtomographic studies. Micron 39:998–1007PubMedCrossRef Libouban H, Blouin S, Moreau MF, Baslé MF, Audran M, Chappard D (2008) Effects of risedronate in a rat model of osteopenia due to orchidectomy and disuse: densitometric, histomorphometric and microtomographic studies. Micron 39:998–1007PubMedCrossRef
44.
go back to reference Kruger MC, Chua WH, Darragh A, Booth CL, Prosser C, Lowry D (2008) Impact of goat milk powdered formulations on mineral absorption, peak bone mass and bone loss due to ovariectomy in rats. J Sci Food Agric 88:1082–1090CrossRef Kruger MC, Chua WH, Darragh A, Booth CL, Prosser C, Lowry D (2008) Impact of goat milk powdered formulations on mineral absorption, peak bone mass and bone loss due to ovariectomy in rats. J Sci Food Agric 88:1082–1090CrossRef
45.
go back to reference Bonjour JP, Benoit V, Payen F, Kraenzlin M (2013) Consumption of yogurts fortified in vitamin D and calcium reduces serum parathyroid hormone and markers of bone resorption: a double-blind randomized controlled trial in institutionalized elderly women. J Clin Endocrinol Metab 98:2015–2021CrossRef Bonjour JP, Benoit V, Payen F, Kraenzlin M (2013) Consumption of yogurts fortified in vitamin D and calcium reduces serum parathyroid hormone and markers of bone resorption: a double-blind randomized controlled trial in institutionalized elderly women. J Clin Endocrinol Metab 98:2015–2021CrossRef
46.
go back to reference Narva M, Rissanen J, Halleen J, Vapaatalo H, Väänänen K, Korpela R (2007) Effects of bioactive peptide, valyl-prolyl-proline (VPP), and lactobacillus helveticus fermented milk containing VPP on bone loss in ovariectomized rats. Ann Nutr Metab 51(1):65–74PubMedCrossRef Narva M, Rissanen J, Halleen J, Vapaatalo H, Väänänen K, Korpela R (2007) Effects of bioactive peptide, valyl-prolyl-proline (VPP), and lactobacillus helveticus fermented milk containing VPP on bone loss in ovariectomized rats. Ann Nutr Metab 51(1):65–74PubMedCrossRef
Metadata
Title
Kefir improves bone mass and microarchitecture in an ovariectomized rat model of postmenopausal osteoporosis
Authors
H.-L. Chen
Y.-T. Tung
C.-H. Chuang
M.-Y. Tu
T.-C. Tsai
S.-Y. Chang
C.-M. Chen
Publication date
01-02-2015
Publisher
Springer London
Published in
Osteoporosis International / Issue 2/2015
Print ISSN: 0937-941X
Electronic ISSN: 1433-2965
DOI
https://doi.org/10.1007/s00198-014-2908-x

Other articles of this Issue 2/2015

Osteoporosis International 2/2015 Go to the issue