Biomechanical and immunohistochemical analysis of high hydrostatic pressure-treated Achilles tendons

Treatment of cancer patients presenting with a solid malignant tumor of the bone or soft tissue requires complete resection of the infiltrated tissue, neoadjuvant chemotherapy, and eventually consecutive reconstruction. Various treatment modalities have been established for the reconstruction of bone and joint defects, including endoprostheses,1 artificial bone substitutes,2 and allogeneic transplants.3 However, each of these reconstruction methods is associated with certain limitations and complications for the patient; thus, new, innovative strategies are strongly desired. The reconstruction of tendons is highly essential for the restoration of joint function, but the refixation of tendons and ligaments to artificial implants is a problematic issue. In this sense, biological reconstruction could help avoid such functional limitation, but allogeneic transplantation of tendons is still a matter of concern, as such a transplant may be contaminated by viruses or bacteria4 and therefore requires special awareness of procurement, storage, and sterilization.5 An alternative approach for limb reconstruction of tumor-affected bone is extracorporal irradiation or autoclaving of resected tissue segments and subsequent reimplantation. For this, it is necessary to devitalize the tumor cells completely to avoid tumor recurrence. Still, with this treatment, destruction of tendons, ligaments, and bone does occur associated with severe loss of biomechanical and biological properties and is therefore a central concern in orthopedic surgery.6–10 Consequently, it is necessary to develop an adequate procedure that would achieve tumor cell inactivation of a bone or tendon without compromising the biomechanical properties of bone, tendons, and ligaments.6–10 In our own recent studies, the effect of high hydrostatic pressure (HHP) on viability of various human tumor-afflicted tissues, tumor cell lines, and normal cells was investigated. At an exposure of 350 MPa Abstract Background. Reconstruction of bone defects caused by malignant tumors is carried out in different ways. At present, tumor-bearing bone segments are devitalized mainly by extracorporeal irradiation or autoclaving, but both methods have substantial disadvantages. In this regard, high hydrostatic pressure (HHP) treatment of the bone is a new, advancing technology that has been used in preclinical testing to inactivate normal cells and tumor cells without altering the biomechanical properties of the bone. The aim of this study was to examine the biomechanical and immunohistochemical properties of tendons after exposure to HHP and to evaluate whether preservation of the bony attachment of tendons and ligaments is possible. Methods. For this, 19 paired Achilles tendons were harvested from both hindlimbs of 4-month-old pigs. After preparation, the cross-sectional area of each tendon was determined by magnetic resonance imaging (MRI). For each animal, one of the two tendons was taken at random and exposed to a pressure of 300 MPa (n = 9) or 600 MPa (n = 10). Results. The contralateral tendon served as an untreated control. The biomechanical properties of the tendons remained unchanged with respect to the tested parameters: Young’s modulus (MPa) and tensile strength (MPa). This finding is in line with immunohistochemical labeling results, as no difference in the labeling pattern of collagen I and versican was observed when comparing the HHP group (at 600 MPa) to the untreated control group. Conclusions. We anticipate that during orthopedic surgery HHP can serve as a novel, promising methodical approach to inactivate Achilles tendon and bone cells without altering the biomechanical properties of the tendons. This should allow one to preserve the attachment of tendon and ligaments to the devitalized bone and to facilitate functional reconstruction.

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