Effect of<i>staphylococcal</i> lipoteichoic acid on differentiation of RAW264.7 cells into osteoclasts _Chinese Journal of Reparative and Reconstructive Surgery

Authors：

RENLirong ¹ ,  XUYongqing ² , WANGHai ² , HEXiaoqing ² , SONGMuguo ² , CHENXueqiu ¹

1. Department of Traumatic Surgery, the Second Affiliated Hospital of Kunming Medical University, Kunming Yunnan, 650101, P.R.China;
2. Department of Orthopedics, Kunming General Hospital of Chengdu Military Command, Kunming Yunnan, 650032, P.R.China;

Corresponding author：

XUYongqing, Email: xuyongqingkm@163.net

Keywords：

Staphylococcal lipoteichoic acid; osteoclasts; RAW264.7 cells; osteomyelitis; bone resorption

DOI：

10.7507/1002-1892.201610077

Video：

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Objective To investigate the effect ofstaphylococcal lipoteichoic acid (LTA-sa) on RAW264.7 cells differentiation into osteoclasts. Methods RAW264.7 cells were cultured with LTA-sa of 100 ng/mL (group A), LTA-sa of 200 ng/mL (group B), LTA-sa of 400 ng/mL (group C), receptor activator of nuclear factor κB (NF-κB) ligand (RANKL) of 100 ng/mL as positive control (group D), and equal volume of PBS as blank control (group E) respectively for 5 days. And then, tartrate resistant acid phosphatase staining (TRAP) was used to detect the formation of osteoclast-like cells, Image-Pro Plus 6.0 software to measure the areas of bone resorption pits in Corning Osteo Assay Surface (COAS) wells, and MTT assay to observe the proliferation activity of RAW264.7 cells in group A, B, C, and E. Results After cultured for 5 days, the formation of osteoclast-like cells and bone resorption pits were observed in all groups. The number of osteoclast-like cells and the area of bone resorption pits in groups A, B, C, and D were more than those in group E. And with the increased concentration of LTA-sa, the indexes in groups A, B, and C increased gradually, but were lower than those in group D, and differences were significant between groups (P<0.05). At 5 days after culture, there was no significant difference in absorbance value among the experimental groups (groups A, B, C, and E) (P>0.05). Conclusion LTA-sa has promoting effect on RAW264.7 cells differentiation into osteoclasts.

Citation： RENLirong, XUYongqing, WANGHai, HEXiaoqing, SONGMuguo, CHENXueqiu. Effect ofstaphylococcal lipoteichoic acid on differentiation of RAW264.7 cells into osteoclasts . Chinese Journal of Reparative and Reconstructive Surgery, 2017, 31(2): 180-184. doi: 10.7507/1002-1892.201610077 Copy

1.	Sims NA, Vrahnas C. Regulation of cortical and trabecular bone mass by communication between osteoblasts, osteocytes and osteoclasts. Arch Biochem Biophys, 2014, 561: 22-28.
2.	Montanaro L, Testoni F, Poggi A, et al. Emerging pathogenetic mechanisms of the implant-related osteomyelitis by Staphylococcus aureus. Int J Artif Organs, 2011, 34(9): 781-788.
3.	Nair SP, Meghji S, Wilson M, et al. Bacterially induced bone destruction: mechanisms and misconceptions. Infect Immun, 1996, 64(7): 2371-2380.
4.	Zecconi A, Scali F. Staphylococcus aureus virulence factors in evasion from innate immune defenses in human and animal diseases. Immunol Lett, 2013, 150(1-2): 12-22.
5.	Wicken AJ, Knox KW. Lipoteichoic acids: a new class of bacterial antigen. Science, 1975, 187(4182): 1161-1167.
6.	Kishimoto T, Kaneko T, Ukai T, et al. Peptidoglycan and lipopolysaccharide synergistically enhance bone resorption and osteoclastogenesis. J Periodontal Res, 2012, 47(4): 446-454.
7.	Zhou C, Liu WG, He W, et al. Saikosaponin a inhibits RANKL-induced osteoclastogenesis by suppressing NF-κB and MAPK pathways. Int Immunopharmacol, 2015, 25(1): 49-54.
8.	Jin T, Zhu YL, Li J, et al. Staphylococcal protein A, Panton-Valentine leukocidin and coagulase aggravate the bone loss and bone destruction in osteomyelitis. Cell Physiol Biochem, 2013, 32(2): 322-333.
9.	Rasigade JP, Trouillet-Assant S, Ferry T, et al. PSMs of hypervirulent Staphylococcus aureus act as intracellular toxins that kill infected osteoblasts. PLoS One, 2013, 8(5): e63176-e63188.
10.	Josse J, Velard F, Gangloff SC. Staphylococcus aureus vs. Osteoblast: Relationship and consequences in osteomyelitis. Fron Cell Infect Microbiol, 2015, 5: 85.
11.	Nair S, Song Y, Meghji S, et al. Surface-associated proteins from Staphylococcus aureus demonstrate potent bone resorbing activity. J Bone Miner Res, 1995, 10(5): 726-734.
12.	Lau YS, Wang W, Sabokbar A, et al. Staphylococcus aureus capsular material promotes osteoclast formation. Injury, 2006, 37 Suppl 2: S41-S48.
13.	Meghji S, Crean SJ, Hill PA, et al. Surface-associated protein from Staphylococcus aureus stimulates osteoclastogenesis: possible role in S.aureus-induced bone pathology. Br J Rheumatol, 1998, 37(10): 1095-1101.
14.	Ryu YH, Baik JE, Yang JS, et al. Differential immunostimulatory effects of Gram-positive bacteria due to their lipoteichoic acids. Int Immunopharmacol, 2009, 9(1): 127-133.
15.	Chambers TJ, Revell PA, Fuller K, et al. Resorption of bone by isolated rabbit osteoclasts. J Cell Sci, 1984, 66: 383-399.
16.	Yang J, Ryu YH, Yun CH, et al. Impaired osteoclastogenesis by staphylococcal lipoteichoic acid through Toll-like receptor 2 with partial involvement of MyD88. J Leukoc Biol, 2009, 86(4): 823-831.
17.	Yang J, Park OJ, Kim J, et al. Lipoteichoic acid of enterococcus faecalis inhibits the differentiation of macrophages into osteoclasts. J Endod, 2016, 42(4): 570-574.
18.	Kobayashi K, Takahashi N, Jimi E, et al. Tumor necrosis factor alpha stimulates osteoclast differentiation by a mechanism Independent of the ODF/RANKL-RANK interaction. J Exp Med, 2000, 191(2): 275-286.
19.	Yao W, Li K, Liao K. Macropinocytosis contributes to the macrophage foam cell formation in RAW264.7 cells. Acta Biochim Biophys Sin (Shanghai), 2009, 41(9): 773-780.
20.	Soysa NS, Alles N, Aoki K, et al. Osteoclast formation and differentiation: an overview. J Med Dent Sci, 2012, 59(3): 65-74.
21.	Collin-Osdoby P, Yu X, Zheng H, et al. RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Med, 2003, 80: 153-166.
22.	任莉荣,徐永清,王海,等. 金黄色葡萄球菌肽聚糖对破骨细胞分化的影响研究.中国修复重建外科杂志, 2016, 30(8): 1006-1010.
23.	Mendoza Bertelli A, Delpino MV, Lattar S, et al. Staphylococcus aureus protein A enhances osteoclastogenesis via TNFR1 and EGFR signaling. Biochim Biophys Acta, 2016, 1862(10): 1975-1983.
24.	任莉荣, 王海, 何晓清,等. 金葡菌脂磷壁酸促进破骨细胞分化的分子机制. 实用医学杂志, 2016, 32(20): 3369-3372.

1. Sims NA, Vrahnas C. Regulation of cortical and trabecular bone mass by communication between osteoblasts, osteocytes and osteoclasts. Arch Biochem Biophys, 2014, 561: 22-28.
2. Montanaro L, Testoni F, Poggi A, et al. Emerging pathogenetic mechanisms of the implant-related osteomyelitis by Staphylococcus aureus. Int J Artif Organs, 2011, 34(9): 781-788.
3. Nair SP, Meghji S, Wilson M, et al. Bacterially induced bone destruction: mechanisms and misconceptions. Infect Immun, 1996, 64(7): 2371-2380.
4. Zecconi A, Scali F. Staphylococcus aureus virulence factors in evasion from innate immune defenses in human and animal diseases. Immunol Lett, 2013, 150(1-2): 12-22.
5. Wicken AJ, Knox KW. Lipoteichoic acids: a new class of bacterial antigen. Science, 1975, 187(4182): 1161-1167.
6. Kishimoto T, Kaneko T, Ukai T, et al. Peptidoglycan and lipopolysaccharide synergistically enhance bone resorption and osteoclastogenesis. J Periodontal Res, 2012, 47(4): 446-454.
7. Zhou C, Liu WG, He W, et al. Saikosaponin a inhibits RANKL-induced osteoclastogenesis by suppressing NF-κB and MAPK pathways. Int Immunopharmacol, 2015, 25(1): 49-54.
8. Jin T, Zhu YL, Li J, et al. Staphylococcal protein A, Panton-Valentine leukocidin and coagulase aggravate the bone loss and bone destruction in osteomyelitis. Cell Physiol Biochem, 2013, 32(2): 322-333.
9. Rasigade JP, Trouillet-Assant S, Ferry T, et al. PSMs of hypervirulent Staphylococcus aureus act as intracellular toxins that kill infected osteoblasts. PLoS One, 2013, 8(5): e63176-e63188.
10. Josse J, Velard F, Gangloff SC. Staphylococcus aureus vs. Osteoblast: Relationship and consequences in osteomyelitis. Fron Cell Infect Microbiol, 2015, 5: 85.
11. Nair S, Song Y, Meghji S, et al. Surface-associated proteins from Staphylococcus aureus demonstrate potent bone resorbing activity. J Bone Miner Res, 1995, 10(5): 726-734.
12. Lau YS, Wang W, Sabokbar A, et al. Staphylococcus aureus capsular material promotes osteoclast formation. Injury, 2006, 37 Suppl 2: S41-S48.
13. Meghji S, Crean SJ, Hill PA, et al. Surface-associated protein from Staphylococcus aureus stimulates osteoclastogenesis: possible role in S.aureus-induced bone pathology. Br J Rheumatol, 1998, 37(10): 1095-1101.
14. Ryu YH, Baik JE, Yang JS, et al. Differential immunostimulatory effects of Gram-positive bacteria due to their lipoteichoic acids. Int Immunopharmacol, 2009, 9(1): 127-133.
15. Chambers TJ, Revell PA, Fuller K, et al. Resorption of bone by isolated rabbit osteoclasts. J Cell Sci, 1984, 66: 383-399.
16. Yang J, Ryu YH, Yun CH, et al. Impaired osteoclastogenesis by staphylococcal lipoteichoic acid through Toll-like receptor 2 with partial involvement of MyD88. J Leukoc Biol, 2009, 86(4): 823-831.
17. Yang J, Park OJ, Kim J, et al. Lipoteichoic acid of enterococcus faecalis inhibits the differentiation of macrophages into osteoclasts. J Endod, 2016, 42(4): 570-574.
18. Kobayashi K, Takahashi N, Jimi E, et al. Tumor necrosis factor alpha stimulates osteoclast differentiation by a mechanism Independent of the ODF/RANKL-RANK interaction. J Exp Med, 2000, 191(2): 275-286.
19. Yao W, Li K, Liao K. Macropinocytosis contributes to the macrophage foam cell formation in RAW264.7 cells. Acta Biochim Biophys Sin (Shanghai), 2009, 41(9): 773-780.
20. Soysa NS, Alles N, Aoki K, et al. Osteoclast formation and differentiation: an overview. J Med Dent Sci, 2012, 59(3): 65-74.
21. Collin-Osdoby P, Yu X, Zheng H, et al. RANKL-mediated osteoclast formation from murine RAW 264.7 cells. Methods Mol Med, 2003, 80: 153-166.
22. 任莉荣,徐永清,王海,等. 金黄色葡萄球菌肽聚糖对破骨细胞分化的影响研究.中国修复重建外科杂志, 2016, 30(8): 1006-1010.
23. Mendoza Bertelli A, Delpino MV, Lattar S, et al. Staphylococcus aureus protein A enhances osteoclastogenesis via TNFR1 and EGFR signaling. Biochim Biophys Acta, 2016, 1862(10): 1975-1983.
24. 任莉荣, 王海, 何晓清,等. 金葡菌脂磷壁酸促进破骨细胞分化的分子机制. 实用医学杂志, 2016, 32(20): 3369-3372.

Chinese Journal of Reparative and Reconstructive Surgery

Effect ofstaphylococcal lipoteichoic acid on differentiation of RAW264.7 cells into osteoclasts

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