Early events associated with bone healing in patients with type 2 diabetes mellitus appear to be delayed. Hyperglycaemia and an associated increase in oxidative stress are cited as potential factors leading to a change in cellular behaviour. Using an in vivo model monitoring bone formation around implants placed into rat mandibles, we have previously identified that the onset of cell proliferation and osteoblast differentiation are delayed and subsequently prolonged compared with normal bone. This study used the same implant model to characterize oxidative stress biomarkers and primary antioxidant enzyme profiles during diabetic bone healing in vivo. Implants were placed into the sockets of incisors extracted from the mandibles of normal Wistar and diabetic Goto-Kakizaki rats for 3 and 9 weeks after implant insertion. Histochemical analysis confirmed a delay in bone healing around implants in diabetic animals. Immunohistochemical localization of peri-cellular staining for protein carbonyl groups, as a biomarker of oxidized protein content, was slightly higher in diabetic granulation tissue compared with normal tissue. However, no differences were observed in the staining patterns of advanced glycation end products. Minimal differences were observed in the number of cells positive for cytoplasmic superoxide dismutase (SOD)1 or mitochondrial SOD2. Significantly, catalase was absent in diabetic tissues. The results suggest that the oxidative environment in healing bone is differentially affected by hyperglycaemia, particularly in relation to catalase. The significance of these observations for diabetic bone healing is discussed.

Keywords:
Reactive oxygen species, Bone, Osseointegration, Type 2 diabetes
1.
Ahmed, N. (2005) Advanced glycation endproducts – role in pathology of diabetic complications. Diabetes Res Clin Pract 67: 3–21.
2.
Aust, S.D., L.A. Morehouse, C.E. Thomas (1985) Role of metals in oxygen radical reactions. J Free Radic Biol Med 1: 3–25.
3.
Bai, X.C., D. Lu, J. Bai, H. Zheng, Z.Y. Ke, X.M. Li, et al. (2004) Oxidative stress inhibits osteoblastic differentiation of bone cells by ERK and NF-ĸB. Biochem Biophys Res Commun 314: 197–207.
4.
Bonnefont-Rousselot, D. (2002) Glucose and reactive oxygen species. Curr Opin Clin Nutr Metab Care 5: 561–568.
5.
Casap, N., S. Nimri, E. Ziv, J. Sela, Y. Samuni (2008) Type 2 diabetes has minimal effect on osseointegration of titanium implants in Psammomys obesus. Clin Oral Implants Res 19: 458–64.
6.
Catherwood, M.A., L.A. Powell, P. Anderson, D. McMaster, P.C. Sharpe, E.R. Trimble (2002) Glucose-induced oxidative stress in mesangial cells. Kidney Int 61: 599–608.
7.
Cavarape, A., F. Feletto, F. Mercuri, L. Quagliaro, G. Daman, A. Ceriello A. (2001) High-fructose diet decreases catalase mRNA levels in rat tissues. J Endocrinol Invest 24: 838–845.
8.
Chen, R.M., G.J. Wu, H.C. Chang, J.T. Chen, T.F. Chen TF, Y.L. Lin, et al. (2005) 2,6-diisopropylphenol protects osteoblasts from oxidative stress-induced apoptosis through suppression of caspase-3 activation. Ann NY Acad Sci 1042: 448–459.
9.
Collins, T., M.I. Cybulsky (2001) NF-ĸB: pivotal mediator or innocent bystander in atherogenesis? J Clin Invest 107: 255–264.
10.
Colombo, J.S., D. Balani, A.J. Sloan, S.J. Crean, J. Okazaki, R.J. Waddington (2010) Delayed osteoblast differentiation and altered inflammatory response around implants placed in incisor sockets of type 2 diabetic rats. Clin Oral Impl Res, E-pub ahead of print.
11.
Dalle-Donne, I., R. Rossi, D. Giustarini, A. Milzani, R. Colombo (2003) Protein carbonyl groups as biomarkers of oxidative stress. Clin Chim Acta 329: 23–38.
12.
Dougan, M., G. Dranoff (2008) Inciting inflammation: the RAGE about tumor promotion. J Exp Med 205: 267–270.
13.
Frank, J., D.K. Kelleher, A. Pompella, O. Thews, H.K. Biesalski, P. Vaupel (1998) Enhancement of oxidative cell injury and antitumor effects of localized 44°C hyperthermia upon combination with respiratory hyperoxia and xanthine oxidase. Cancer Res 58: 2693–269.
14.
Fujii, H., Y. Hamada, M. Fukagawa (2008) Bone formation in spontaneously diabetic Torii – newly established model of non-obese type 2 diabetes rats. Bone 42: 372–379.
15.
Góth, L., J.W. Eaton (2000) Hereditary catalase deficiencies and increased risk of diabetes. Lancet 356: 1820–1821.
16.
Goto, Y., M. Kakizaki, N. Masaki (1975) Spontaneous diabetes produced by selective breeding of normal Wistar rats. Proc Natl Jpn Acad 51: 80–85.
17.
Graves, D.T., R. Liu, T.W. Oates (2007) Diabetes-enhanced inflammation and apoptosis: impact on periodontal pathosis. Periodontol 2000 45: 128–137.
18.
Hamada, Y., H. Fujii, M. Fukagawa (2009) Role of oxidative stress in diabetic bone disorder. Bone 44: 936–941.
19.
Hamanaka, R.B., N.S. Chandel (2010) Mitochondrial reactive oxygen species regulate cellular signaling and dictate biological outcomes. Trends Biochem Sci 35: 505–513.
20.
Hasegawa, H., S. Ozawa, K. Hashimoto, T. Takeichi, T. Ogawa (2008) Type 2 diabetes impairs implant osseointegration capacity in rats. Int J Oral Maxillofac Implants 23: 237–246.
21.
Houslay, M.D. (1991) ‘Crosstalk’: a pivotal role for protein kinase C in modulating relationships between signal transduction pathways. Eur J Biochem 195: 9–27.
22.
Inoguchi, T., P. Li, F. Umeda, H.Y. Yu, M. Kakimoto, M. Imamura, et al. (2000) High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells. Diabetes 49: 1939–1945.
23.
Kowluru, R.A., P.S. Chan (2007) Oxidative stress and diabetic retinopathy. Exp Diabetes Res 2007: 43603.
24.
Koya, D., G.L. King (1998) Protein kinase C activation and the development of diabetic complications. Diabetes 47: 859–866.
25.
Kume, S., S. Kato, S.I. Yamagishi, Y. Inagaki, S. Ueda, N. Arima, et al. (2005) Advanced glycation end-products attenuate human mesenchymal stem cells and prevent cognate differentiation into adipose tissue, cartilage and bone. J Bone Miner Res 20: 1647–1658.
26.
Lu, H., D. Kraut, L.C. Gerstenfeld, D.T. Graves (2003) Diabetes interferes with the bone formation by affecting the expression of transcription factors that regulate osteoblast differentiation. Endocrinology 144: 346–352.
27.
Nishikawa, T., D. Edelstein, X.L. Du, S. Yamagishi, T. Matsumura, et al. (2000) Normalizing mitochondrial superoxide production blocks three pathways of hyperglycaemic damage. Nature 404: 787–790.
28.
Sadi, G., O. Yilmaz, T. Güray T (2008) Effect of vitamin C and lipoic acid on streptozotocin-induced diabetes gene expression: mRNA and protein expressions of Cu-Zn SOD and catalase. Mol Cell Biochem 309: 109–116.
29.
Sakai, D., J. Okazaki, Y. Komasa (2008) Bone healing of tooth extracted socket in type 2 diabetes. J Oral Tissue Eng 5: 134–144.
30.
Santana, R.B., L. Xu, H.B. Chase, S. Amar, D.T. Graves, P.C. Trackman (2003) A role for advanced glycation end products in diminished bone healing in type 1 diabetes. Diabetes 52: 1502–1510.
31.
Sheweita, S.A., K.I. Khoshhal (2007) Calcium metabolism and oxidative stress in bone fractures: role of antioxidants. Curr Drug Metab 8: 519–525.
32.
Srivastava, A.K. (2002) High glucose-induced activation of protein kinase signaling pathways in vascular smooth muscle cells: a potential role in the pathogenesis of vascular dysfunction in diabetes. Int J Mol Med 9: 85–89.
33.
Stolzing, A., N. Coleman, A. Scutt (2006) Glucose-induced replicative senescence in mesenchymal stem cells. Rejuvenation Res 9: 31–35.
34.
Treins, C., S. Giorgetti-Peraldi, J. Murdaca, E. Van Obberghen (2001) Regulation of vascular endothelial growth factor expression by advanced glycation end products. J Biol Chem 276: 43836–43841.
35.
Waddington, R.J., R. Moseley, G. Embery (2000) Reactive oxygen species: a potential role in the pathogenesis of periodontal diseases. Oral Dis 6: 138–151.
36.
Zhen, D., Y. Chen, X. Tang (2010) Metformin reverses the deleterious effects of high glucose on osteoblast function. J Diabetes Complications 24: 334–344.
© 2011 S. Karger AG, Basel
2011
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.