Oral Biofilm And Osteoarthritis - More Than Just A Planktonic Relationship?
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1 Oral Biofilm And Osteoarthritis - More Than Just A Planktonic Relationship? Joanne E. Haughan, Mag.med.vet 1, Noreen J. Hickock, PhD 2, Irving Shapiro, PhD 2, Javad Parvizi, M.D. 3, Garth Ehrlich, PhD 4, Thomas P. Schaer, VMD 1. 1 University of Pennsylvania, Comparative Orthopaedic Research Laboratory, Kennett Square, PA, USA, 2 Thomas Jefferson University, Philadelphia, PA, USA, 3 The Rothman Institute, Philadelphia, PA, USA, 4 Center for Genome Science, Pittsburgh, PA, USA. Disclosures: J.E. Haughan: None. N.J. Hickock: None. I. Shapiro: None. J. Parvizi: None. G. Ehrlich: None. T.P. Schaer: None. Introduction: Degenerative orthopedic conditions such as osteoarthritis (OA) are linked to a loss of mobility and often cause extreme and chronic pain. Annual costs in terms of lost work time and treatment exceed $100 billion. Most authorities would agree that non-surgical interventions are inadequate while surgical responses have varied outcomes including complications such as debilitating infections. Accordingly, there is an urgent need for a new approach to treating this disease. A recent study demonstrated a direct correlation between knee OA and dental pathogens 1 and ongoing research indicates that over 75% of primary knee surgeries show contamination by pathogens, many of which are facultative or frank anaerobes commonly resident in the oral cavity. Bacteria from oral biofilm can enter the blood stream in their planktonic form via compromised periodontal tissue. We hypothesize that normal aging in the presence of periodontal disease predisposes the hypoxic joint tissue to colonization by mobilized anaerobic oral pathogens; this colonization triggers inflammatory cascades that promote and/or accelerate OA. Methods: In an animal model pilot study approved by the IACUC of the University of Pennsylvania we analyzed articular cartilage and synovial tissue biopsies from femoro-tibial joints as well as samples of sub- and supra-gingival plaque and crevicular fluid from three female Yucatan miniature pigs aged 6-9 years. Cranial and bilateral hindlimb CT examinations were performed on each animal under general anesthesia. Animals were then placed into ventral recumbency and a thorough dental exam was performed Gingival bleeding upon probing was scored from 0-3, periodontal pocket depth was measured in millimeters using a calibrated dental probe and samples of crevicular fluid and sub- and supragingival plaque were obtained from multiple sites throughout the oral cavity. The animals were repositioned into dorsal recumbency and both stifles were aseptically prepared. A minimally invasive femoro-tibial joint arthrotomy was performed bilaterally via a lateral para-patellar incision. Synovial tissue and cartilage samples were obtained using Ferris-Smith rongeurs. Samples were placed into sterile saline, flash frozen in liquid nitrogen and stored at -80 degrees until analysis. Both oral and articular samples were analyzed using the Ibis T5000 Universal Biosensor which uses mass spectrometry derived base composition signatures obtained from PCR amplification of bacterial 16S rdna to identify organisms present in a sample 2. Presence of select microbes in cartilage samples was verified using Fluorescent In Situ Hybridization (FISH). Results: All three pigs had severe calculus accumulation (figure 1), gingival bleeding upon probing (figure 2) and evidence of tooth loss and alveolar bone resorption were visible on cranial CT examinations (figure 3). The average gingival bleeding index was 1.9 and average periodontal pocket depths were between 3.9 and 5.1mm (Range: 3-16mm) where >3mm is defined as pathological (table 1). The results from dental exams indicated that all three animals had severe, advanced periodontal disease. Mild osteoarthritic changes were seen in the femoro-tibial joints of all three animals on preoperative hind limb CT images. A significant number of predominantly anaerobic or facultative anaerobic microorganisms were identified in plaque and crevicular fluid samples as well as in femoro-tibial joint cartilage and synovial tissue from all animals (tables 2&3). Presence of P. gingivalis, S. auricularis and S. suis in cartilage samples was verified using Fluorescent In Situ Hybridization (FISH). Discussion: Microorganisms associated with compromised oral health can be isolated from degenerative joint tissue in miniature pigs with naturally occurring periodontal disease, suggesting that bacterial colonization of the knee with the ensuing initiation of inflammatory cascades could be a mechanism for initiation of degenerative changes associated with arthritis. We have recently expanded our ongoing research to
2 include samples of intervertebral disc tissue as well as endplate cartilage and bone and are developing a translational miniature pig model using an established technique to induced periodontal disease 3 in order to study how compromised oral health influences the initiation and progression of trauma-induced IVDD and OA Significance: Our research could offer new mechanistic associations between poor oral health and degenerative orthopedic disease. Acknowledgments: The authors would like to thank Rachel Kreft of the Center for Genomic Science for her work on microbiological testing, as well as the animal care staff and research assitants at Penn Vet CORL for their assistance with animal surgeries and care. References: 1. Temoin, S., Chakaki, A., Askari, A., El-Halaby, A., Fitzgerald, S., Marcus, R.E., Han, Y.W., and Bissada, N.F. Identification of oral bacterial DNA in synovial fluid of patients with arthritis with native and failed prosthetic joints. J. Clin. Rhematol.18: , Ecker, D.J., Drader, J.J., Gutierrez, J., Gutierrez, A., Hannis, J.C., Schink, A., Sampath,R., Blyn, L.B., Eshoo, M. W., Hall, T. A., Tobarmosquera, M., Jiang, Y., Sannes-Lowery, K.A., Cummins, L.L., Libby, B., Walcott, D.M., Massire, C., Ranken, R., Manalili, S., Ivy, C., Melton, R., Levene, H., Harpin, V., Li, F., White, N., Pear, M., Ecker, J.A., Samant, V., Knize, D., Robbins, D., Rudnick, K., Hajjar, F., Hofstadler, S.A. The Ibis T5000 Universal Biosensor: An Automated Platform for Pathogen Identification and Strain Typing JALA 2006: 11; Hickey, J.S., O Neal, R.B., Scheidt, M.J., Strong, S.L., Turgeon, D., Van Dyke, T.E. Microbiologic Characterization of Ligature- Induced Peri-Implantitis in the Micro-Swine Model. Journal of Periodontology 1991: 62 (9);
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