Boo Yong Sim; Ji Won Bak; Hae Jin; Ji Ae Jun; Hak Joo Choi; Chang Ju Kwon; Hwa Young Kim; Kevin J. Ruff; Karsten Brandt; Dong Hee Kim

doi:10.4163/jnh.2015.48.4.310

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Sim, Bak, Jin, Jun, Choi, Kwon, Kim, Ruff, Brandt, and Kim: Effects of natural eggshell membrane (NEM) on monosodium iodoacetate-induced arthritis in rats∗

Research Article

J Nutr Health 2015;48(4):310-318.

Published online: 19 January 2015

DOI: https://doi.org/10.4163/jnh.2015.48.4.310

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

Boo Yong Sim¹, Ji Won Bak¹, Hae Jin¹, Ji Ae Jun¹, Hak Joo Choi¹, Chang Ju Kwon², Hwa Young Kim², Kevin J. Ruff³, Karsten Brandt⁴, Dong Hee Kim^1,^†

¹Traditional and Biomedical Research Center (TBRC-RIC), Daejeon University, Daejeon 300-176, Korea

²Ju Yeong NS Co., Ltd., Seoul 137-872, Korea

³Scientific & Regulatory Affairs, ESM Technologies, LLC, 2213 Missouri St. 64836, USA

⁴Human Nutrition, Stratum Nutrition, 43204, Tarragona, Spain

^†To whom correspondence should be addressed. tel: +82-42-280-2623, e-mail: dhkim@dju.kr

^∗ This work was (partly) supported by the RIC program of MOTIE (Ministry of Trade, Industry and Energy) in Daejeon University.

Received 7 May 2015 Revised 9 June 2015 Accepted 29 June 2015

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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-1β, and IL-6 were measured by Luminex and PGE_2, MMP-2, MMP-9, TIMP-1, LTB_4, 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-1β, IL-6, PGE_2, 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.

Keywords: osteoarthritis, anti-Inflammatory, NEM, MIA, health food

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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.

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.

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.

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.

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.

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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