Lack of cytocompatibility in bone substitutes impairs healing in surrounding bone. Adverse biological events around biomaterials may be associated with oxidative stress. We hypothesized that a clinically used inorganic bone substitute is cytotoxic to osteoblasts due to oxidative stress and that N-acetyl cysteine (NAC), an antioxidant amino acid derivative, would detoxify such material. Only 20% of rat calvaria osteoblasts were viable when cultured on commercial deproteinized bovine bone particles for 24 hr, whereas this percentage doubled on bone substitute containing NAC. Intracellular ROS levels markedly increased on and under bone substitutes, which were reduced by prior addition of NAC to materials. NAC restored suppressed alkaline phosphatase activity in the bone substitute. Proinflammatory cytokine levels from human osteoblasts on the bone substitute decreased by one-third or more with addition of NAC. NAC alleviated cytotoxicity of the bone substitute to osteoblastic viability and function, implying enhanced bone regeneration around NAC-treated inorganic biomaterials.
Get full access to this article
View all access options for this article.
References
1.
Accorsi-Mendonca T, Conz MB, Barros TC, de Sena LA, Soares Gde A, Granjeiro JM (2008). Physicochemical characterization of two deproteinized bovine xenografts. Braz Oral Res22:5–10.
2.
Alpar B, Leyhausen G, Gunay H, Geurtsen W (2000). Compatibility of resorbable and nonresorbable guided tissue regeneration membranes in cultures of primary human periodontal ligament fibroblasts and human osteoblast-like cells. Clin Oral Investig4:219–225.
3.
Att W, Yamada M, Kojima N, Ogawa T (2009). N-acetyl cysteine prevents suppression of oral fibroblast function on poly(methylmethacrylate) resin. Acta Biomater5:391–398.
4.
Baggiolini M, Dewald B, Moser B (1994). Interleukin-8 and related chemotactic cytokines—CXC and CC chemokines. Adv Immunol55:97–179.
5.
Bertazzo S, Bertran CA (2008). Effect of hydrazine deproteination on bone mineral phase: a critical view. J Inorg Biochem102:137–145.
6.
Carter DH, Scully AJ, Heaton DA, Young MP, Aaron JE (2002). Effect of deproteination on bone mineral morphology: implications for biomaterials and aging. Bone31:389–395.
7.
Dent P, Yacoub A, Fisher PB, Hagan MP, Grant S (2003). MAPK pathways in radiation responses. Oncogene22:5885–5896.
8.
Feinendegen LE (2002). Reactive oxygen species in cell responses to toxic agents. Hum Exp Toxicol21:85–90.
9.
Gillissen A, Nowak D (1998). Characterization of N-acetylcysteine and ambroxol in anti-oxidant therapy. Respir Med92:609–623.
10.
Gillissen A, Bartling A, Schoen S, Schultze-Werninghaus G (1997). Antioxidant function of ambroxol in mononuclear and polymorphonuclear cells in vitro. Lung175:235–242.
11.
Huang FM, Tsai CH, Yang SF, Chang YC (2005). Induction of interleukin-6 and interleukin-8 gene expression by root canal sealers in human osteoblastic cells. J Endod31:679–683.
12.
Ignjatovic N, Ninkov P, Kojic V, Bokurov M, Srdic V, Krnojelac D, et al. (2006). Cytotoxicity and fibroblast properties during in vitro test of biphasic calcium phosphate/poly-dl-lactide-co-glycolide biocomposites and different phosphate materials. Microsc Res Tech69:976–982.
13.
Ishimi Y, Miyaura C, Jin CH, Akatsu T, Abe E, Nakamura Y, et al. (1990). IL-6 is produced by osteoblasts and induces bone resorption. J Immunol145:3297–3303.
14.
Kaplan SS, Basford RE, Jeong MH, Simmons RL (1994). Mechanisms of biomaterial-induced superoxide release by neutrophils. J Biomed Mater Res28:377–386.
15.
Keegan GM, Learmonth ID, Case CP (2008). A systematic comparison of the actual, potential, and theoretical health effects of cobalt and chromium exposures from industry and surgical implants. Crit Rev Toxicol38:645–674.
16.
Kojima N, Yamada M, Paranjpe A, Tsukimura N, Kubo K, Jewett A, et al. (2008). Restored viability and function of dental pulp cells on polymethylmethacrylate (PMMA)-based dental resin supplemented with N-acetyl cysteine (NAC). Dent Mater24:1686–1693.
17.
Lenz R, Mittelmeier W, Hansmann D, Brem R, Diehl P, Fritsche Y, et al. (2009). Response of human osteoblasts exposed to wear particles generated at the interface of total hip stems and bone cement. J Biomed Mater Res A89:370–378.
18.
Liu Q, Cen L, Yin S, Chen L, Liu G, Chang Y, et al. (2008). A comparative study of proliferation and osteogenic differentiation of adipose-derived stem cells on akermanite and beta-TCP ceramics. Biomaterials29: 4792–4799.
19.
Luo X, Budihardjo I, Zou H, Slaughter C, Wang X (1998). Bid, a Bcl2 interacting protein, mediates cytochrome c release from mitochondria in response to activation of cell surface death receptors. Cell94:481–490.
20.
Nadzam GS, De La Cruz C, Greco RS, Haimovich B (2000). Neutrophil adhesion to vascular prosthetic surfaces triggers nonapoptotic cell death. Ann Surg231:587–599.
21.
Paranjpe A, Cacalano NA, Hume WR, Jewett A (2007). N-acetylcysteine protects dental pulp stromal cells from HEMA-induced apoptosis by inducing differentiation of the cells. Free Radic Biol Med43:1394–1408.
22.
Patel JD, Krupka T, Anderson JM (2007). iNOS-mediated generation of reactive oxygen and nitrogen species by biomaterial-adherent neutrophils. J Biomed Mater Res A80:381–390.
23.
Rauner M, Sipos W, Pietschmann P (2007). Osteoimmunology. Int Arch Allergy Immunol143:31–48.
24.
Schweikl H, Spagnuolo G, Schmalz G (2006). Genetic and cellular toxicology of dental resin monomers. J Dent Res85:870–877.
25.
Somanathan RV, Simunek A (2006). Evaluation of the success of beta-tricalciumphosphate and deproteinized bovine bone in maxillary sinus augmentation using histomorphometry: a review. Acta Medica (Hradec Kralove)49:87–89.
26.
Spagnuolo G, D’Anto V, Cosentino C, Schmalz G, Schweikl H, Rengo S (2006). Effect of N-acetyl-L-cysteine on ROS production and cell death caused by HEMA in human primary gingival fibroblasts. Biomaterials27:1803–1809.
27.
Stefaniak A, Leonard S, Hoover M, Virji M, Day G (2009). Dissolution and reactive oxygen species generation of inhaled cemented tungsten carbide particles in artificial human lung fluids. J Physics: Conference Series151(012045). DOI: 10.1088/1742-6596/151/1/012045.
28.
Tadic D, Epple M (2004). A thorough physicochemical characterisation of 14 calcium phosphate-based bone substitution materials in comparison to natural bone. Biomaterials25:987–994.
29.
Taylor JC, Cuff SE, Leger JP, Morra A, Anderson GI (2002). In vitro osteoclast resorption of bone substitute biomaterials used for implant site augmentation: a pilot study. Int J Oral Maxillofac Implants17:321–330.
30.
Tsaryk R, Kalbacova M, Hempel U, Scharnweber D, Unger RE, Dieter RE, et al. (2007). Response of human endothelial cells to oxidative stress on Ti6Al4V alloy. Biomaterials28:806–813.
31.
Tsukimura N, Yamada M, Aita H, Hori N, Yoshino F, Chang-Il Lee M, et al. (2009). N-acetyl cysteine (NAC)-mediated detoxification and functionalization of poly(methyl methacrylate) bone cement. Biomaterials30:3378–3389.
32.
Vermes C, Chandrasekaran R, Jacobs JJ, Galante JO, Roebuck KA, Glant TT (2001). The effects of particulate wear debris, cytokines, and growth factors on the functions of MG-63 osteoblasts. J Bone Joint Surg Am83(A):201–211.
33.
Yamada M, Ogawa T (2009). Chemodynamics underlying N-acetyl cysteine-mediated bone cement monomer detoxification. Acta Biomater5: 2963–2973.
34.
Yamada M, Kojima N, Paranjpe A, Att W, Aita H, Jewett A, et al. (2008). N-acetyl cysteine (NAC)-assisted detoxification of PMMA resin. J Dent Res87:372–377.
35.
Yamada M, Kojima N, Att W, Hori N, Suzuki T, Ogawa T (2009). N-acetyl cysteine restores viability and function of rat odontoblast-like cells impaired by polymethylmethacrylate dental resin extract. Redox Rep14:13–22.
36.
Zafarullah M, Li WQ, Sylvester J, Ahmad M (2003). Molecular mechanisms of N-acetylcysteine actions. Cell Mol Life Sci60:6–20.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
