J Nutr Health.  2015 Aug;48(4):310-318. 10.4163/jnh.2015.48.4.310.

Effects of natural eggshell membrane (NEM) on monosodium iodoacetate-induced arthritis in rats

• Sim BY
• Bak JW
• Lee HJ
• Jun JA
• Choi HJ
• Kwon CJ
• Kim HY
• Ruff KJ
• Brandt K
• Kim DH

• KMID: 2209897
• DOI: http://doi.org/10.4163/jnh.2015.48.4.310

Abstract

PURPOSE
The aim of this study is to investigate anti-arthritis activity using natural eggshell membrane (NEM).
METHODS
NEM was administered at 52 mg/kg, 200 mg/kg, and 400 mg/kg to SD-Rat, where arthritis was induced by monosodium iodoacetate (MIA) at 3 mg. NO production in serum was measured using Griess reagent. Cytokines including IL-1beta, and IL-6 were measured by Luminex and PGE2, MMP-2, MMP-9, TIMP-1, LTB4, and hs-CRP were measured by ELISA. The cartilage of patella volume was examined and 3-D high-resolution reconstructions of the cartilage of patella were obtained using a Micro-CT system.
RESULTS
Production of NO, IL-1beta, IL-6, PGE2, MMP-2, MMP-9, TIMP-1, LTB4, and hs-CRP in serum was decreased, respectively, in comparison with control. The cartilage of patella volume increased significantly. In addition, the NEM group showed a decrease in the cartilage of patella, synovial membrane, and transformation of fibrous tissue.
CONCLUSION
The results for NEM showed significant anti-arthritis activity. These results may be developed as a raw material for new health food to ease the symptoms mentioned above.

Keyword

osteoarthritis; anti-Inflammatory; NEM; MIA; health food

MeSH Terms

Animals
Arthritis*
Cartilage
Cytokines
Dinoprostone
Enzyme-Linked Immunosorbent Assay
Food, Organic
Interleukin-6
Leukotriene B4
Membranes*
Osteoarthritis
Patella
Rats*
Synovial Membrane
Tissue Inhibitor of Metalloproteinase-1
Cytokines
Dinoprostone
Interleukin-6
Leukotriene B4
Tissue Inhibitor of Metalloproteinase-1

Figure

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  Fig. 1. Effects of NEM on levels of NO in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗∗∗p < 0.001). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

• 

  Fig. 2. Effects of NEM on levels of PGE2 in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗p < 0.05, ∗∗p < 0.01). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/ kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

• 

  Fig. 3. Effects of NEM on levels of IL-1ß, IL-6 in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA -induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

• 

  Fig. 4. Effects of NEM on levels of hs-CRP in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗∗∗p < 0.001). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/ kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

• 

  Fig. 5. Effects of NEM on levels of MMP-2, MMP-9 in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗p < 0.05, ∗∗p < 0.01). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA (3 mg/mL)-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

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  Fig. 6. Effects of NEM on levels of LTB4 in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗∗p < 0.01, ∗∗∗p < 0.001). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

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  Fig. 7. Effects of NEM on levels of COMP in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗p < 0.05). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

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  Fig. 8. Effects of NEM on levels of CTX-II in the serum of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗∗p < 0.01). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

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  Fig. 9. Effects of NEM on joint pathology (Hematoxylin & Eosin staining) and (Safranin-O) from joint tissue of MIA-induced osteoarthritis rats. Normal group shows the presence of slightly thickened synovium (arrow) Control group shows isolated areas with chronic inflammation (arrow). NEM treated group shows signs of tissue integrity with a thick layer of cartilage compared with control group (arrow). Normal, Normal SD-rat group; Control, MIA - induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

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  Fig. 10. Effects of NEM on imaging of cartilage degeneration using micro CT-arthrography in joint tissue of MIA-induced osteoarthritis rat. The results were expressed as mean ± SD from 7 osteoarthritis rats. Statistically significant value compared with control group by unpaired student's t-test (∗p < 0.05). Normal, Normal SD-rat group; Control, MIA-induced osteoarthritis group; 52, MIA-induced osteoarthritis group + NEM 52 mg/kg; 200, MIA-induced osteoarthritis group + NEM 200 mg/kg; 400, MIA-induced osteoarthritis group + NEM 400 mg/kg.

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