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Chitosan scaffolds and copper\u2010chitosan scaffolds were fabricated and characterized by scanning electron microscopy (SEM). Chitosan and copper\u2010chitosan scaffolds were implanted into 5 mm diameter critical\u2010sized calvarial defects in Fisher 344 male rats. Empty defects (no scaffolds) were included as a control. After 4 weeks, the rats were sacrificed for microcomputed tomography (micro\u2010CT) and histological analysis of new bone tissue development. Microscopy images revealed the uniformly porous structure of chitosan and copper\u2010chitosan scaffolds. Significant bone regeneration was noted in the defects treated with copper\u2010chitosan scaffolds when evaluated using micro\u2010CT and histological analysis, when compared with other groups tested. On analysis of the micro\u2010CT scans, an eleven\u2010fold and a two\u2010fold increase in the new bone volume\/total volume (BV\/TV) % was found in defects treated with the copper\u2010chitosan scaffolds, when compared to empty defects and chitosan scaffolds, respectively. This study demonstrated the suitability of copper\u2010crosslinked chitosan scaffolds for bone tissue engineering and provides the first evidence that inclusion of copper ions in scaffolds can enhance tissue regeneration. \u00a9 2014 Wiley Periodicals, Inc. 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