Technical Note Polyhexanide and hydrogen peroxide inhibit proteoglycan synthesis of human chondrocytes Eric Ro¨hner1, Paula Hoff2, Tobias Winkler1, Philipp von Roth 1 , Jo¨rn Bengt Seeger1, Carsten Perka 1 , Georg Matziolis1 1Department of Traumatology and Orthopaedics, and 2Department of Rheumatology and Clinical Immunology, Charite´ - Universita¨tsmedizin, Berlin, Germany The use of local antiseptics is a common method in septic joint surgery. We tested polyhexanide and hydrogen peroxide, two of the most frequently used antiseptics with high efficacy and low toxicity. The purpose of this study was to evaluate the effects of both antiseptics on the extracellular cartilaginous matrix synthesis of human chondrocytes. Chondrocytes were isolated from donated human knee joints, embedded in alginate beads, and incubated for 10 and 30 minutes with polyhexanide (0.04%), hydrogen peroxide (3%), or phosphate-buffered saline (PBS) for control. Cartilaginous matrix production was quantified through light microscopic analysis of Alcian blue staining. Cell number and morphology were detected by histological analysis. Chondrocytes showed a decreased intensity of blue colouring after antiseptic treatment versus PBS. In contrast to that, neither the cell number per view field nor the cell morphology differed between the groups. Polyhexanide has more toxic potential than hydrogen peroxide. Based on the fact that the cell number and morphology was not altered by the substances at the examined concentrations, the lower intensity of Alcian blue staining of treated chondrocytes indicates a decreased cartilage-specific matrix synthesis by polyhexanide more than by hydrogen peroxide and control. Keywords: Antiseptics, Human chondrocytes, Hydrogen peroxide, Polyhexanide, Proteoglycans Introduction Different studies showed that shoulder and knee joint infections often result from inﬁltrations with local anaesthetics, glucocorticoids, or hyaluronic acid con- trary to hip joint infections which are rarely inﬁltrated for diagnostic or therapeutic reasons.1–3 In addition to arthroscopic or open debridement, the application of local antiseptics is a standard procedure in the treatment of joint infections. The most frequently used antiseptics of local joint and wound infections are polyhexanide, and hydrogen peroxide. In contrast to antibiotic substances, they do not differ between procariontic and eukaryontic cells. Therefore, the tissue toxicity of antiseptic substances has to be determined for every tissue coming into contact. There is an ongoing discussion between elimination of bacteria and toxicity against chondrocytes.4,5 It should be assumed that the time of chondrocytes Correspondence to: Eric Ro¨hner, Center for Musculoskeletal Surgery, Orthopaedic Department, Charite´ University Hospital, Berlin, Charite´platz 1, 10117 Berlin, Germany. Email: eric.roehner@charite.de contact with local antiseptics should be between 10 and 30 minutes to eliminate most colonies of bacteria and though to limit chondrocyte toxicity. The most frequent bacteria, which are responsible for wound and joint infections, are staphylococci, streptococci, and Gram-negative bacteria.1,6–8 Be- sides inducing cell death, antiseptic substances may alter the cartilage-speciﬁc synthesis of proteoglycan in chondrocytes and thereby affect the cartilage tissue. The undisturbed interaction between the chondrocytes and extracellular matrix plays an im- portant role for the regulation of chondrocyte homeostasis, which is important for the mainte- nance, the development, and the stabilisation of chondrocyte phenotype.9–12 Consequently, an inter- ruption of proteoglycan synthesis results in cartilage damage with the loss of chondrocytes.9,13 Two of the most commonly used antiseptics in septic joint surgery are polyhexanide and hydrogen peroxide. Literature reviews show insufﬁcient data about the effects of antiseptics on human cartilage. 2011 National Society for Histotechnology MORE OpenChoice articles are open access and distributed under the terms of the Creative Commons Attribution License 3.0 DOI 10.1179/014788811X12949268296121 Journal of Histotechnology 2011 VOL 34 NO 1 35