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      Remodelling of human hamstring autografts after anterior cruciate ligament reconstruction

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          Abstract

          Purpose

          Histological analysis of the remodelling process of human hamstring tendon (HT) grafts after standardized anterior cruciate ligament reconstruction (ACLR) with an accelerated rehabilitation protocol.

          Methods

          Sixty-seven patients underwent retrieval of mid-substance biopsies after clinically successful hamstring autograft ACLR. Samples were allocated to one of three groups depending on the time point of retrieval: group 1 (6–12 months; n = 15), group 2 (13–24 months; n = 16) and group 3 (>24 months; n = 11).

          Biopsies from native HT ( n = 17) and ACL ( n = 8) served as controls. Cellular density, vascular density and myofibroblast density and collagen fibril alignment were analysed by haematoxylin–eosin, Masson-Goldner-Trichrom and immunohistochemical staining protocols.

          Results

          Compared with native HT (330.4/mm²), total cell number was increased in groups 1-3 (Group 1 = 482.0/mm² ( P = 0.036); group 2 = 850.9/mm² ( P = 0.005); and group 3 = 595.6/mm² ( P = 0.043). There were no significant differences between the groups for vessel density. Myofibroblast density was higher in group 2 (199.6/mm²) compared with native HT (1.9/mm², P = 0.014). Collagen orientation was irregular up to 12 months. Thereafter, collagen orientation became more regular, adapting to, but not fully restoring, the appearance of the intact ACL. For the first 12 months, cells were predominantly ovoid. Ensuing cell morphology changed to spindle shaped in group 2 and predominantly narrow long cells over 24 months.

          Conclusion

          Human hamstring grafts showed typical stages of graft remodelling, which was not complete up to 2 years after ACLR. The remodelling process in humans was prolonged compared with the results obtained in several animal studies.

          Level of evidence

          Case–control study, Level III.

          Related collections

          Most cited references29

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          Biomechanical analysis of an anatomic anterior cruciate ligament reconstruction.

          The focus of most anterior cruciate ligament reconstructions has been on replacing the anteromedial bundle and not the posterolateral bundle. Anatomic two-bundle reconstruction restores knee kinematics more closely to normal than does single-bundle reconstruction. Controlled laboratory study. Ten cadaveric knees were subjected to external loading conditions: 1) a 134-N anterior tibial load and 2) a combined rotatory load of 5-N x m internal tibial torque and 10-N x m valgus torque. Resulting knee kinematics and in situ force in the anterior cruciate ligament or replacement graft were determined by using a robotic/universal force-moment sensor testing system for 1) intact, 2) anterior cruciate ligament deficient, 3) single-bundle reconstructed, and 4) anatomically reconstructed knees. Anterior tibial translation for the anatomic reconstruction was significantly closer to that of the intact knee than was the single-bundle reconstruction. The in situ force normalized to the intact anterior cruciate ligament for the anatomic reconstruction was 97% +/- 9%, whereas the single-bundle reconstruction was only 89% +/- 13%. With a combined rotatory load, the normalized in situ force for the single-bundle and anatomic reconstructions at 30 degrees of flexion was 66% +/- 40%and 91% +/- 35%, respectively. Anatomic reconstruction may produce a better biomechanical outcome, especially during rotatory loads. Results may lead to the use of a two-bundle technique. Copyright 2002 American Orthopaedic Society for Sports Medicine
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            Ligamentization of tendon grafts treated with an endogenous preparation rich in growth factors: gross morphology and histology.

            To investigate whether the application of a particular platelet-rich plasma preparation rich in growth factors (PRGF) during anterior cruciate ligament (ACL) surgery gives a potential advantage for better tendon graft ligamentization. This study included 37 volunteers who underwent either conventional (control group, n = 15) or PRGF-assisted (n = 22) ACL reconstruction with an autogenous hamstring and required second-look arthroscopy to remove hardware or loose bodies, treat meniscal tears or plica syndrome, or resect cyclops lesions at 6 to 24 months after ACL surgery. The gross morphologies of the grafts were evaluated on second-look arthroscopy by use of the full arthroscopic score (0 to 4 points) to evaluate graft thickness and apparent tension (0 to 2 points) plus synovial coverage (0 to 2 points). At the same time, biopsy specimens were harvested uniformly from the grafted tendons. In these specimens the histologic transformation of the tendon graft to ACL-like tissue was evaluated by use of the Ligament Tissue Maturity Index, and a score to assess the progression of new connective tissue enveloping the graft was created by use of 3 criteria previously used to characterize changes during ligament healing: cellularity, vascularity, and collagen properties. The overall arthroscopic evaluation of PRGF-treated grafts showed an excellent rating in 57.1% of the knees (score of 4) and a fair rating in 42.9% (score of 2 or 3). In contrast, evaluation of untreated grafts showed an excellent rating in 33.3% of the knees, a fair rating in 46.7%, and a poor rating in 20% (score of 0 or 1). Overall, arthroscopic evaluations were not statistically different between PRGF and control groups (P = .051). PRGF treatment influenced the histologic characteristics of the tendon graft, resulting in tissue that was more mature than in controls (P = .024). Histologically evident newly formed connective tissue enveloping the graft was present in 77.3% of PRGF-treated grafts and 40% of controls. The appearance of the connective tissue envelope changed with increasing time from surgery. On the basis of the histologic findings, we suggest that the remodeling of PRGF-treated grafts involves the formation of synovial-like tissue enveloping the graft. This tissue is eventually integrated in the remodeled tendon graft, conferring a similar appearance to the normal ACL. The use of PRGF influenced the histologic characteristics of tendon grafts, resulting in more remodeling compared with untreated grafts. We have shown temporal histologic changes during the 6- to 24-month postoperative period of graft maturation, with newly formed connective tissue enveloping most grafts treated with PRGF. Level III, case-control study. Copyright 2010 Arthroscopy Association of North America. Published by Elsevier Inc. All rights reserved.
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              Tendon healing in a bone tunnel. Part II: Histologic analysis after biodegradable interference fit fixation in a model of anterior cruciate ligament reconstruction in sheep.

              Tendon-to-bone healing of soft-tissue grafts has been described to progress by the development of a fibrous interzone that undergoes a maturation process leading to the development of an indirect type of ligament insertion. Previous studies used extra-articular models or fixation far away from the joint line; thus, no data are available investigating tendon-to-bone healing of a soft-tissue graft fixed anatomically. Therefore, we studied the tendon-to-bone healing of the anatomic soft-tissue graft interference fit fixation in a model of anterior cruciate ligament (ACL) reconstruction in sheep. Animal study. Thirty-five mature sheep underwent ACL reconstruction with an autologous Achilles tendon split graft. Grafts were directly fixed with biodegradable poly-(D,L-lactide) interference screws. Animals were euthanized after 6, 9, 12, 24, and 52 weeks and histologic evaluations were performed. Undecalcified specimens were evaluated under normal and polarized light. Additionally, animals received a polychrome sequential labeling (tetracycline, xylenol orange, and calcein green) to determine bone growth per time under fluorescent light. Intratunnel histologic findings at 6 weeks showed a tendon-bone junction with only a partial fibrous interzone between the graft tissue and the surrounding bone. A mature intratunnel tendon-bone junction with a zone of fibrocartilage was found at 9 to 12 weeks. At the tunnel entrance site a wide regular ligamentous insertion site was seen in all specimens after 24 weeks. This insertion showed regular patterns such as the direct type of insertion of a normal ligament with a dense basophilic transition zone consisting of mineralized cartilage. A fibrous interzone between the graft tissue and the bone tunnel was only partially developed, which is in contrast to all previous studies in which nonanatomic fixation was used. Thus, it is reasonable to assume that the tendon-to-bone healing in the present study may progress partially by direct-contact healing without the development of a fibrous interzone. To our knowledge, this is the first report describing the development of a direct type of ligament insertion after ACL replacement with a soft-tissue graft. This is in contrast to previous studies reporting the development of an indirect type of insertion when using nonanatomic fixation far away from the joint line. Thus, histologic data strongly indicate that anatomic interference fit fixation is beneficial for tendon-to-bone incorporation by leading to the development of a direct type of ligament insertion.
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                Author and article information

                Contributors
                r.janssen@mmc.nl , www.rpajanssen.com
                Journal
                Knee Surg Sports Traumatol Arthrosc
                Knee Surgery, Sports Traumatology, Arthroscopy
                Springer-Verlag (Berlin/Heidelberg )
                0942-2056
                1433-7347
                4 February 2011
                4 February 2011
                August 2011
                : 19
                : 8
                : 1299-1306
                Affiliations
                [1 ]Orthopaedic Center Máxima, Máxima Medical Center, Postbus 90052, 5600 PD Eindhoven, The Netherlands
                [2 ]Department of Orthopaedic Surgery and Traumatology, Center for Musculoskeletal Surgery, Charité, Campus Mitte, University Medicine Berlin, Charité Platz 1, 10117 Berlin, Germany
                Article
                1419
                10.1007/s00167-011-1419-y
                3136699
                21293848
                50bc62b4-227c-4703-9f52-f17c7aafaf05
                © The Author(s) 2011
                History
                : 21 July 2010
                : 24 January 2011
                Categories
                Knee
                Custom metadata
                © Springer-Verlag 2011

                Surgery
                anterior cruciate ligament reconstruction,remodelling,myofibroblast,human,hamstring autograft

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