Limits...
Pomegranate Peel Extract Prevents Bone Loss in a Preclinical Model of Osteoporosis and Stimulates Osteoblastic Differentiation in Vitro.

Spilmont M, Léotoing L, Davicco MJ, Lebecque P, Miot-Noirault E, Pilet P, Rios L, Wittrant Y, Coxam V - Nutrients (2015)

Bottom Line: The nutritional benefits of pomegranate have attracted great scientific interest.Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia.In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers.

View Article: PubMed Central - PubMed

Affiliation: Unité de Nutrition Humaine, CRNH Auvergne, UMR 1019, INRA, F-63000 Clermont-Ferrand, France. mel.spilmont@gmail.com.

ABSTRACT
The nutritional benefits of pomegranate have attracted great scientific interest. The pomegranate, including the pomegranate peel, has been used worldwide for many years as a fruit with medicinal activity, mostly antioxidant properties. Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia. In this study, with in vivo and ex vivo experiments, we investigated whether the consumption of pomegranate peel extract (PGPE) could limit the process of osteopenia. We demonstrated that in ovariectomized (OVX) C57BL/6J mice, PGPE consumption was able to significantly prevent the decrease in bone mineral density (-31.9%; p < 0.001 vs. OVX mice) and bone microarchitecture impairment. Moreover, the exposure of RAW264.7 cells to serum harvested from mice that had been given a PGPE-enriched diet elicited reduced osteoclast differentiation and bone resorption, as shown by the inhibition of the major osteoclast markers. In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers. PGPE may be effective in preventing the bone loss associated with ovariectomy in mice, and offers a promising alternative for the nutritional management of this disease.

No MeSH data available.


Related in: MedlinePlus

Osteoclast viability, differentiation and transcription in culture after exposure to serum harvested from mice either raised under control conditions or treated with pomegranate peel extract. * p < 0.05 PGPE versus Control condition. (A) Cell proliferation test performed on RAW264.7 cell line cultured in minimal medium (2% Fetal Bovine Serum, FBS), supplemented with serum from mice exposed to either physiological serum (1.25% FBS + 0.75% Control mice serum) or pomegranate peel extract, PGPE (1.25% FBS + 0.75 PGPE mice serum) after 1 and 2 days of treatment. Results are expressed as mean ± standard error of the mean (SEM) (n = 6). (B) Tartrate resistant acid phosphatase, TRAP activity of RAW264.7 cells cultured in optimized medium with 10% FBS or with 7.5% FBS + 2.5% mice serum from animals fed either physiological serum (Control) or pomegranate peel extract (PGPE), after 3 days of treatment. Results are expressed as mean ± SEM (n = 8). (C) Transcriptomic analysis of the main osteoclast markers in RAW264.7 cells cultured in optimized medium after 2 days of treatment with serum from mice exposed to either physiological serum (Control) or pomegranate peel extract (PGPE). Transcriptomic analysis of the Raw264.7 mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to Control condition (fold change = 1) as mean ± standard deviation (SD) (n = 6). Gene: CaM: calmodulin; CCR2: chemokine receptor 2; CTR: calcitonin receptor; CTSK: cathepsin K; FOS; IFN β1: interferon β1; MMP9: metalloproteinase 9; Myc, RANK: receptor activator of nuclear factor-κB; Relb; c-SRC: and TRAF6.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4663593&req=5

nutrients-07-05465-f003: Osteoclast viability, differentiation and transcription in culture after exposure to serum harvested from mice either raised under control conditions or treated with pomegranate peel extract. * p < 0.05 PGPE versus Control condition. (A) Cell proliferation test performed on RAW264.7 cell line cultured in minimal medium (2% Fetal Bovine Serum, FBS), supplemented with serum from mice exposed to either physiological serum (1.25% FBS + 0.75% Control mice serum) or pomegranate peel extract, PGPE (1.25% FBS + 0.75 PGPE mice serum) after 1 and 2 days of treatment. Results are expressed as mean ± standard error of the mean (SEM) (n = 6). (B) Tartrate resistant acid phosphatase, TRAP activity of RAW264.7 cells cultured in optimized medium with 10% FBS or with 7.5% FBS + 2.5% mice serum from animals fed either physiological serum (Control) or pomegranate peel extract (PGPE), after 3 days of treatment. Results are expressed as mean ± SEM (n = 8). (C) Transcriptomic analysis of the main osteoclast markers in RAW264.7 cells cultured in optimized medium after 2 days of treatment with serum from mice exposed to either physiological serum (Control) or pomegranate peel extract (PGPE). Transcriptomic analysis of the Raw264.7 mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to Control condition (fold change = 1) as mean ± standard deviation (SD) (n = 6). Gene: CaM: calmodulin; CCR2: chemokine receptor 2; CTR: calcitonin receptor; CTSK: cathepsin K; FOS; IFN β1: interferon β1; MMP9: metalloproteinase 9; Myc, RANK: receptor activator of nuclear factor-κB; Relb; c-SRC: and TRAF6.

Mentions: With regard to the osteoclast model (RAW264.7 cells), proliferation was significantly increased at day 2 in all conditions, in comparison to day 1. However, mouse serum induced a slight decrease in osteoclast proliferation on the first culture day compared to what was observed in the FBS condition. No significant difference was noticed between the PGPE and Control serum conditions (−36% and −21% respectively; p < 0.5) (Figure 3A). Then, on day 2, we observed that in the PGPE condition, osteoclast proliferation was reduced compared to the Control mouse serum condition (−26%, p < 0.01).


Pomegranate Peel Extract Prevents Bone Loss in a Preclinical Model of Osteoporosis and Stimulates Osteoblastic Differentiation in Vitro.

Spilmont M, Léotoing L, Davicco MJ, Lebecque P, Miot-Noirault E, Pilet P, Rios L, Wittrant Y, Coxam V - Nutrients (2015)

Osteoclast viability, differentiation and transcription in culture after exposure to serum harvested from mice either raised under control conditions or treated with pomegranate peel extract. * p < 0.05 PGPE versus Control condition. (A) Cell proliferation test performed on RAW264.7 cell line cultured in minimal medium (2% Fetal Bovine Serum, FBS), supplemented with serum from mice exposed to either physiological serum (1.25% FBS + 0.75% Control mice serum) or pomegranate peel extract, PGPE (1.25% FBS + 0.75 PGPE mice serum) after 1 and 2 days of treatment. Results are expressed as mean ± standard error of the mean (SEM) (n = 6). (B) Tartrate resistant acid phosphatase, TRAP activity of RAW264.7 cells cultured in optimized medium with 10% FBS or with 7.5% FBS + 2.5% mice serum from animals fed either physiological serum (Control) or pomegranate peel extract (PGPE), after 3 days of treatment. Results are expressed as mean ± SEM (n = 8). (C) Transcriptomic analysis of the main osteoclast markers in RAW264.7 cells cultured in optimized medium after 2 days of treatment with serum from mice exposed to either physiological serum (Control) or pomegranate peel extract (PGPE). Transcriptomic analysis of the Raw264.7 mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to Control condition (fold change = 1) as mean ± standard deviation (SD) (n = 6). Gene: CaM: calmodulin; CCR2: chemokine receptor 2; CTR: calcitonin receptor; CTSK: cathepsin K; FOS; IFN β1: interferon β1; MMP9: metalloproteinase 9; Myc, RANK: receptor activator of nuclear factor-κB; Relb; c-SRC: and TRAF6.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4663593&req=5

nutrients-07-05465-f003: Osteoclast viability, differentiation and transcription in culture after exposure to serum harvested from mice either raised under control conditions or treated with pomegranate peel extract. * p < 0.05 PGPE versus Control condition. (A) Cell proliferation test performed on RAW264.7 cell line cultured in minimal medium (2% Fetal Bovine Serum, FBS), supplemented with serum from mice exposed to either physiological serum (1.25% FBS + 0.75% Control mice serum) or pomegranate peel extract, PGPE (1.25% FBS + 0.75 PGPE mice serum) after 1 and 2 days of treatment. Results are expressed as mean ± standard error of the mean (SEM) (n = 6). (B) Tartrate resistant acid phosphatase, TRAP activity of RAW264.7 cells cultured in optimized medium with 10% FBS or with 7.5% FBS + 2.5% mice serum from animals fed either physiological serum (Control) or pomegranate peel extract (PGPE), after 3 days of treatment. Results are expressed as mean ± SEM (n = 8). (C) Transcriptomic analysis of the main osteoclast markers in RAW264.7 cells cultured in optimized medium after 2 days of treatment with serum from mice exposed to either physiological serum (Control) or pomegranate peel extract (PGPE). Transcriptomic analysis of the Raw264.7 mRNA levels determined by Taqman Low density Arrays are presented as fold change compared to Control condition (fold change = 1) as mean ± standard deviation (SD) (n = 6). Gene: CaM: calmodulin; CCR2: chemokine receptor 2; CTR: calcitonin receptor; CTSK: cathepsin K; FOS; IFN β1: interferon β1; MMP9: metalloproteinase 9; Myc, RANK: receptor activator of nuclear factor-κB; Relb; c-SRC: and TRAF6.
Mentions: With regard to the osteoclast model (RAW264.7 cells), proliferation was significantly increased at day 2 in all conditions, in comparison to day 1. However, mouse serum induced a slight decrease in osteoclast proliferation on the first culture day compared to what was observed in the FBS condition. No significant difference was noticed between the PGPE and Control serum conditions (−36% and −21% respectively; p < 0.5) (Figure 3A). Then, on day 2, we observed that in the PGPE condition, osteoclast proliferation was reduced compared to the Control mouse serum condition (−26%, p < 0.01).

Bottom Line: The nutritional benefits of pomegranate have attracted great scientific interest.Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia.In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers.

View Article: PubMed Central - PubMed

Affiliation: Unité de Nutrition Humaine, CRNH Auvergne, UMR 1019, INRA, F-63000 Clermont-Ferrand, France. mel.spilmont@gmail.com.

ABSTRACT
The nutritional benefits of pomegranate have attracted great scientific interest. The pomegranate, including the pomegranate peel, has been used worldwide for many years as a fruit with medicinal activity, mostly antioxidant properties. Among chronic diseases, osteoporosis, which is associated with bone remodelling impairment leading to progressive bone loss, could eventually benefit from antioxidant compounds because of the involvement of oxidative stress in the pathogenesis of osteopenia. In this study, with in vivo and ex vivo experiments, we investigated whether the consumption of pomegranate peel extract (PGPE) could limit the process of osteopenia. We demonstrated that in ovariectomized (OVX) C57BL/6J mice, PGPE consumption was able to significantly prevent the decrease in bone mineral density (-31.9%; p < 0.001 vs. OVX mice) and bone microarchitecture impairment. Moreover, the exposure of RAW264.7 cells to serum harvested from mice that had been given a PGPE-enriched diet elicited reduced osteoclast differentiation and bone resorption, as shown by the inhibition of the major osteoclast markers. In addition, PGPE appeared to substantially stimulate osteoblastic MC3T3-E1 alkaline phosphatase (ALP) activity at day 7, mineralization at day 21 and the transcription level of osteogenic markers. PGPE may be effective in preventing the bone loss associated with ovariectomy in mice, and offers a promising alternative for the nutritional management of this disease.

No MeSH data available.


Related in: MedlinePlus