Injury to the anterior cruciate ligament (ACL) often results in functional instability within the knee joint and may induce a severe articular deterioration. Clinically, these patients may require an ACL reconstruction. In an attempt to develop an improved ligament prosthesis, a prototype xenograft--an epoxy-fixed porcine Achilles tendon--was developed. The study was intended to investigate the crosslinking characteristics of the epoxy-fixed porcine tendons fixed at different pHs (4.0, 7.4, 9.0, and 10.5), temperatures (4, 25, and 37 degrees C), and fixative concentrations (1, 2, or 4%). Samples of each group were taken out at various elapsed fixation periods. The crosslinking characteristics-fixation index, denaturation temperature, and moisture content-of each sample were determined. Additionally, the stiffness of the epoxy-fixed tendons under various fixation conditions was qualitatively compared. In the study it was noted that the color of porcine Achilles tendons remained natural after epoxy fixation. With increasing pH, temperature, or fixative concentration, the fixation indices and denaturation temperatures of the epoxy-fixed tendons increased. However, the fixed porcine tendons tended to be stiffer at a higher pH, temperature, or fixative concentration. The moisture contents of the epoxy-fixed tendons were relatively lower than the fresh ones, while they were approximately the same at different pHs, temperatures, and fixative concentrations. The implications of these findings for the epoxy-fixed porcine tendons in ACL reconstruction require further investigation.

Anterior cruciate ligament (ACL) injuries, if left untreated, often produce significant disability in the athletically active population. Currently, autogenous tissue is the most commonly used substitute for ACL reconstruction because its immunogenicity is virtually nonexistent. However, the functional amount of autogenous tissue available for transplantation is limited. Additionally, this transplantation procedure may create a defect at the donor site, which can result in functional disability. To address these concerns, a prototype xenograft ligament prosthesis, epoxy-fixed porcine Achilles tendon, was developed. This study was intended to investigate the crosslinking characteristics of the epoxy-fixed porcine tendon. The fresh and glutaraldehyde-fixed porcine Achilles tendons were used as controls. Fresh porcine Achilles tendons procured from a slaughterhouse were used to fabricate the ligament prostheses. A 4% epoxy (ethylene glycol diglycidyl ether) solution or a 0.625% glutaraldehyde solution was employed to fix the porcine tendons. Samples of each group were taken out at various elapsed fixation periods. The crosslinking characteristics- denaturation temperature, moisture content, and fixation index-of each sample were then determined. In the study, it was learned that the crosslinking rate for the glutaraldehyde fixation was faster than that for the epoxy fixation. While the denaturation temperatures and the fixation indices for both studied groups were higher than for the fresh one, the denaturation temperature of the glutaraldehyde-fixed tendon was relatively higher than its epoxy-fixed counterpart. However, the fixation index and the moisture content for both studied groups were comparable. Also, it was noted that the epoxy-fixed tendon appeared more natural as compared to its glutaraldehyde-fixed counterpart. The implications of these findings for the epoxy-fixed tendon in the clinical ACL reconstruction require further investigation.

The purpose of this study was to investigate the in vitro degradation potential of porcine pericardia fixed with various aldehyde or epoxy compound (EC) fixatives, using bacterial collagenase and pronase. The fixatives investigated were formaldehyde (FA), glutaraldehyde (GA), monofunctional EC (EX-131), and multifunctional ECs (EX-810, EX-313, and EX-512). Fresh porcine pericardium was used as a control. The test samples were well immersed in a 20-U/mL collagenase solution or a 10-U/mL pronase solution and incubated at 37 degrees C at pH 7.5 for 24 h. The extent of degradation of each test sample was determined by measuring its increment in free amino group content and changes in collagen structure, denaturation temperature, and tensile stress after degradation. In general, the extent of tissue degradation with pronase was more notable than with collagenase. As observed with fresh tissue, the EX-131 EC fixed tissue radically disintegrated after either collagenase or pronase degradation, whereas the other test samples remained intact. The reason for this may reside in the more random molecular packing of the EX-131 EC-fixed tissue, which led to some loss in its helical integrity. This made penetration of enzymes into biological tissue easier. Of the multifunctional EC test groups, tissues fixed with tetrafunctional EC (EX-521) or trifunctional EC (EX-313) had relatively better resistance to degradation than those fixed with bifunctional EC (EX-810). The extent of degradation for the EX-313 or EX-512 EC fixed tissues was similar to that observed for the FA- or GA-fixed tissues. The results of this study indicated that the biological tissue fixed with monofunctional EC (EX-131) cannot resist bacterial collagenase or pronase degradation. However, resistance to degradation of the multifunctional EC (EX-313 or EX-152)-fixed tissues was comparable to that of the aldehyde (FA or GA)-fixed tissues. Therefore, of various EC fixatives, the EC with a greater number of functional groups should be chosen for tissue fixation to increase its resistance to enzymatic degradation.

Due to the nature of bioprostheses, which are mainly biological tissues that cannot be sterilized with heat or irradiation, the sterilization method by choice is generally liquid chemicals. It is known that a number of liquid chemicals can have rapid germicidal effect and can be used to sterilize bioprostheses. The study was to evaluate the effects of various chemical sterilization methods on the crosslinking and enzymatic degradation characteristics of an epoxy-fixed biological tissue. The chemical sterilants employed were: a 70% ethanol solution (EtOH), a 2% epoxy compound + 20% ethanol solution (EX-810), a 2% propylene oxide + 20% ethanol solution (PO), and a 0.625% glutaraldehyde + 20% ethanol + 0.2% polysorbate solution (GA). Both masking and crosslinking of the free amino groups within the epoxy-fixed tissue were observed subsequent to sterilization with GA or EX-810. This improved the resistance of the GA or EX-810 sterilized tissues against collagenase degradation as compared to its nonsterilized counterpart. However, subsequent to sterilization with PO, only masking of the free amino groups within the epoxy-fixed tissue was noted. The inhibition of the collagenase degradation by masking of the free amino groups was traded off by the more random molecular packing of the PO sterilized sample due to the introduction of the side branches. Sterilization of the epoxy-fixed tissue with EtOH may increase its denaturation temperature and tensile strength, while neither masking nor crosslinking of free amino groups within the tissue took place. The resistance to degradation of the EtOH sterilized tissue, however, did not improve as compared to its nonsterilized counterpart.

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