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Tenocytes from Ruptured and Tendinopathic Achilles Tendons Produce Greater Quantities of Type III Collagen than Tenocytes from Normal Achilles Tendons

An In Vitro Model of Human Tendon Healing

Nicola Maffulli, MD, MS, PhD, FRCS(Orth)^*,, Stanley W.B. Ewen, PhD, FRCPath, Stuart W. Waterston, BSc Med Sci (Hons)^*, Jacqueline Reaper, BSc Med Sci (Hons)^* and Victoria Barrass, BSc Med Sci (Hons)^*

^* Department of Orthopaedic Surgery, University of Aberdeen, Aberdeen, Scotland
Department of Pathology, University of Aberdeen, Aberdeen, Scotland

Address correspondence and reprint requests to Nicola Maffulli, MD, Department of Orthopaedic Surgery, University of Aberdeen Medical School, Polwarth Building, Foresterhill, Aberdeen AB25 2ZD, Scotland

Type I collagen is the main collagen in tendons; type III collagen is present in small amounts. Ruptured Achilles tendons contain a significantly greater proportion of type III collagen, which predisposes them to rupture. We used an in vitro model to determine whether tenocytes from Achilles tendons that were ruptured (N = 22), nonruptured (N = 7), tendinopathic (N = 12), and fetal (N = 8) show different behavior. Samples of Achilles tendon were digested with collagenase and the released tenocytes were collected. Primary tenocyte cultures were established and subsequently cultured onto glass coverslips. Once a confluent monolayer was obtained, the cell populations were "wounded" by scraping a pipette tip along the surface. The cultures were further incubated for either 1, 4, 8, 12, 16, or 24 hours, and production of types I and II collagen was assessed by immunostaining. In cultures from ruptured and tendinopathic tendons, there was increased production of type III collagen. Athletic participation places excess stress on the Achilles tendon, which could potentially lead to areas of microtrauma within the tendon. These areas may heal by the production of type III collagen, which is an abnormal healing response. Accumulation of such episodes of microtrauma could result in a critical point where the resistance of the tissue to tensile forces is compromised and tendon rupture occurs.

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