The effect of anti-inflammatory and antifibrotic agents on fibroblasts obtained from arthrofibrotic tissue: An in vitro and in vivo study.

OBJECTIVES: The aims of this study were to determine whether the administration of anti-inflammatory and antifibrotic agents affect the proliferation, viability, and expression of markers involved in the fibrotic development of the fibroblasts obtained from arthrofibrotic tissue in vitro, and to evaluate the effect of the agents on arthrofibrosis prevention in vivo. METHODS: Dexamethasone, diclofenac, and decorin, in different concentrations, were employed to treat fibroblasts from arthrofibrotic tissue (AFib). Cell proliferation was measured by DNA quantitation, and viability was analyzed by Live/Dead staining. The levels of procollagen type I N-terminal propeptide (PINP) and procollagen type III N-terminal propeptide (PIIINP) were evaluated with enzyme-linked immunosorbent assay (ELISA) kits. In addition, the expressions of fibrotic markers were detected by real-time polymerase chain reaction (PCR). Fibroblasts isolated from healthy tissue (Fib) served as control. Further, a rabbit model of joint contracture was used to evaluate the antifibrotic effect of the three different agents. RESULTS: Dexamethasone maintained the viability and promoted the proliferation of AFib. Diclofenac decreased the viability and inhibited the cell proliferation during the first week of cultivation. However, decorin inhibited AFib proliferation and downregulated the expressions of fibrotic markers. Additionally, decorin could improve the flexion contracture angle and inhibit the deposition of interstitial matrix components in the rabbit joint model. CONCLUSION: Decorin decreased the expression of myofibroblast markers in AFib, inhibited the proliferation of AFib, and prevented the initial procedure of arthrofibrosis in vivo, suggesting that decorin could be a promising treatment to inhibit the development of arthrofibrosis.Cite this article: X. Tang, S. Teng, M. Petri, C. Krettek, C. Liu, M. Jagodzinski. The effect of anti-inflammatory and antifibrotic agents on fibroblasts obtained from arthrofibrotic tissue: An in vitro and in vivo study. Bone Joint Res 2018;7:213-222. DOI: 10.1302/2046-3758.73.BJR-2017-0219.R2.

In vivo heterotopic culturing of prefabricated tendon grafts with mechanical stimulation in a sheep model.

BACKGROUND: The goal of this study is to investigate the biomechanical and histological properties of in vivo heterotopically prefabricated cruciate ligament replacement grafts with and without mechanical stimulation. The clinical goal is to heterotopically prefabricate a bone-tendon-bone graft for anterior cruciate ligament reconstruction, which allows rapid ingrowth and early full weight bearing. METHODS: In a sheep model, eight quadriceps tendon grafts were harvested and introduced into culture chambers at their proximal and distal ends. In group S, four tendon-chamber constructs were mechanically stimulated by direct attachment to the quadriceps tendon and patella. In group NS, the same constructs were cultured without proximal attachment. All sheep were sacrificed six weeks postoperatively and the constructs were examined biomechanically and histologically. The healthy contralateral ACL and quadriceps tendon were used as controls. RESULTS: Macroscopically, no obvious ossification could be observed at the ends of the tendon-chamber constructs six weeks postoperatively. Histologically, the tendon tissue from the mechanically stimulated constructs revealed higher counts of cells and capillaries. However, there was less regular cell distribution and collagen fiber orientation compared to the control group. In addition, osteoblasts and osteogenesis were observed in the prefabricated constructs both with and without mechanical stimulation. Biomechanically, there were no significant differences in stiffness, elongation and ultimate failure load between the groups. CONCLUSION: In vivo heterotopic culture of prefabricated tendon grafts may have the potential to stimulate osteoblasts and induce osteogenesis. Future studies with longer follow-up and modifications of the surgical technique and culture conditions are desirable.

[Prefabrication of bone transplants]. / Präfabrikation von Knochentransplantaten.

Prefabrication of bone transplants is a promising option for large defects of the long bones, especially if there is compromised vascularization of the defect. This is especially true for postinfection bone defects and other types of atrophic nonunion. The generation of a foreign body membrane (Masquelet's technique) has been investigated in order to ameliorate the response of the host tissue surrounding the defect. In an experimental animal study, a blood vessel within a bone construct could be used to generate customized, vascularized osteogenic constructs that can be used to treat large bone defects in the future.

Suture anchor repair yields better biomechanical properties than transosseous sutures in ruptured quadriceps tendons.

PURPOSE: This human cadaveric study compares the biomechanical properties of quadriceps tendon repair with suture anchors and the commonly applied transosseous sutures. The hypothesis was that suture anchors provide at least equal results concerning gap formation and ultimate failure load compared with transosseous suture repair. METHODS: Thirty human cadaveric knees underwent tenotomy followed by repair with either 5.5-mm-double-loaded suture anchors [titanium (TA) vs. resorbable hydroxyapatite (HA)] or transpatellar suture tunnels using No. 2 Ultrabraid™ and the Krackow whipstitch. Biomechanical analysis included pretensioning the constructs with 20 N for 30 s and then cyclic loading of 250 cycles between 20 and 100 N at 1 Hz in a servohydraulic testing machine with measurement of elongation. Ultimate failure load analysis and failure mode analysis were performed subsequently. RESULTS: Tendon repairs with suture anchors yielded significantly less gap formation during cyclic loading (20th-250th cycle: TA 1.9 ± 0.1, HA 1.5 ± 0.5, TS 33.3 ± 1.9 mm, p < 0.05) and resisted significantly higher ultimate failure loads (TA 740 ± 204 N, HA 572 ± 67 N, TS 338 ± 60 N, p < 0.05) compared with transosseous sutures. Common failure mode was pull-out of the eyelet within the suture anchor in the HA group and rupture of the suture in the TA and TS group. CONCLUSION: Quadriceps tendon repair with suture anchors yields significantly better biomechanical results than the commonly applied transosseous sutures in this human cadaveric study. These biomechanical findings may change the future clinical treatment for quadriceps tendon ruptures. Randomised controlled clinical trials are desirable for the future. LEVEL OF EVIDENCE: Not applicable, controlled laboratory human cadaveric study.

[Ligament bracing--augmented cruciate ligament sutures: biomechanical studies of a new treatment concept]. / "Ligament bracing"--die augmentierte Kreuzbandnaht: Biomechanische Grundlagen für ein neues Behandlungskonzept.

BACKGROUND: In the context of acute knee dislocations, suture repair of ruptured cruciate ligaments leads to good clinical results in 80% of cases. Disadvantages are low primary stability and subsequently secondary elongation of the sutured ligaments. In the present study, we compared primary stability of suture repair, reinforced by different suture augments, to cruciate ligament reconstruction. OBJECTIVE: The concept of ligament bracing with transosseous suture repair of the cruciate ligaments and additional suture augmentation is biomechanically superior to cruciate ligament reconstruction. MATERIAL AND METHODS: A total of 42 porcine knee joints divided into seven groups were examined. The stability of four different suture/augmentation combinations were compared to cruciate ligament reconstruction with human hamstring tendons. The investigational setup consisted of testing 1000 cycles with 20 N to 154 N load in a.-p. translation and 60° flexion. Elongation and load to failure were measured. RESULTS: Neither reconstruction (3.13 ± 1.65 mm; 362 ± 51 N) nor augmented suture repair (1.89-2.5 mm; 464-624 N) achieved the primary stability of the intact cruciate ligament (0.63 ± 0.34 mm, 1012 ± 91 N). In comparison to ligament reconstruction, all four augmented suture repairs showed minor elongation in the cyclic test and a higher load to failure. The isolated suture repair showed poor results (6.79 ± 4.86 mm, 177 ± 73 N). CONCLUSION: Augmented suture repair provides significantly higher stability compared with isolated suture repair and reconstruction with hamstring tendons. The concept of ligament bracing could be a promising future treatment option in acute knee dislocations. Clinical results remain to be seen.

En bloc prefabrication of vascularized bioartificial bone grafts in sheep and complete workflow for custom-made transplants.

The aim of this pilot study was to determine, in a new experimental model, whether complex bioartificial monoblocs of relevant size and stability can be prefabricated in a defined three-dimensional design, in which the latissimus dorsi muscle serves as a natural bioreactor and the thoracodorsal vessel tree is prepared for axial construct perfusion. Eighteen sheep were included in the study, with six animals in each of three experimental groups. Vitalization of the ß-tricalcium phosphate-based constructs was performed by direct application of unmodified osteogenic material from the iliac crest (group A), in vivo application of nucleated cell concentrate (NCC) from bone marrow aspirate (group B), and in vitro cultivation of bone marrow stromal cells (BMSC) in a perfusion bioreactor system (group C). The contours of the constructs were designed digitally and transferred onto the bioartificial bone grafts using a titanium cage, which was bent over a stereolithographic model of the defined subvolume intraoperatively. At the end of the prefabrication process, only the axial vascularized constructs of group A demonstrated vital bone formation with considerable stability. In groups B and C, the applied techniques were not able to induce ectopic bone formation. The presented computer-assisted workflow allows the prefabrication of custom-made bioartificial transplants.

Biomechanical properties of femoral posterior cruciate ligament fixations.

PURPOSE: The success of reconstructions of the posterior cruciate ligament (PCL) mainly depends on the fixation strength of the tendon-bone interface. Reliable data about the mechanical characteristics of PCL fixation techniques are sparse. The aim of this study was to investigate the biomechanical properties of different femoral PCL fixation techniques. METHODS: Fresh human cadaver quadriceps (Q) and hamstring (H) tendons were harvested and fixed into porcine femora with a press-fit fixation suturing the tendon over a bone bridge (group A), a novel implant post-fixation (group B) or an interference screw fixation (group C). Each group consisted of 10 specimens. The constructs were cyclically stretched and eventually loaded until failure. Elongation during cyclic loading, stiffness, failure mode and maximum failure load was evaluated. RESULTS: Elongation during cyclical loading was significantly larger between the 1st and the 20th cycle than between the 20th and the 500th cycle in all groups (p < 0.05). Maximum failure load was 409 ± 71 (336-517) N in group QA, 456 ± 58 (347-510) N in group QB, 548 ± 116 (400-798) N in group QC, 472 ± 114 N (316-676 N) in group HA, 494 ± 98 N (371-668 N) in group HB and 498 ± 87 N (391-687 N) in group HC (significantly higher for QB compared to QA, p < 0.05). CONCLUSION: This is the first study investigating the biomechanical properties of femoral PCL fixations. Implant-free fixation techniques like press-fit or post-fixations are able to withstand equal biomechanical forces compared to interference screw fixation. The novel fixations described in this study can be considered as a reliable alternative for the reconstruction of PCL using either hamstring or quadriceps tendons.

Toll-like receptor 2/6-dependent stimulation of mesenchymal stem cells promotes angiogenesis by paracrine factors.

Reconstruction of critical size bone defects represents a major challenge in orthopaedic surgery. Insufficient angiogenesis is a limiting factor for engraftment of large-scale tissue transplants. Transplantation or stimulation of local mesenchymal stem cells (MSCs) represents a potential solution to enhance angiogenesis. We recently identified angiogenic properties for the Toll-like receptor (TLR) 2/6 agonist MALP-2 and now investigated if MALP-2 could be used to stimulate MSCs in order to promote angiogenesis in vitro and in vivo. Human MSCs from the bone marrow of healthy subjects were isolated, cultured and expanded in vitro and were shown to be positive for mesenchymal stem cells markers as well as for the MALP-2 receptors TLR2 and TLR6. MALP-2 directly enhanced migration but not proliferation of human MSCs. Conditioned medium from MALP-2 stimulated MSCs significantly increased proliferation, migration and tube formation of endothelial cells. Analysis of the conditioned medium from MSCs revealed that MALP-2 stimulation enhanced the secretion of several chemokines and growth factors including vascular endothelial growth factors (VEGF) and granulocyte-macrophage colony-stimulating factor (GM-CSF). Finally, we studied MALP-2 effects on MSCs in a sheep model of tissue engineering in vivo. Therefore, MSCs were isolated from the iliac crest of black head sheep and co-cultivated with MALP-2 ex vivo. Implantation of autologous MSCs within a scaffold cylinder into the M. latissimus dorsi significantly enhanced vessel density of these constructs after 6 months. We here present the first evidence that TLR2/6-dependent stimulation of MSCs promotes angiogenesis in vitro and in vivo offering a novel strategy for therapeutic angiogenesis, e.g., for tissue engineering of bone.

[Intracondylar segment osteotomy: correction of intra-articular malalignment after fracture of the tibial plateau]. / Intrakondyläre Segmentosteotomie: Korrektur intraartikulärer Fehlstellungen nach Tibiakopffraktur.

Intracondylar deformities after tibial plateau fractures are intra-articular deformities which present within the medial and/or lateral knee compartments. They exist either isolated or in combination with other extra-articular deformities and/or ligament problems. These deformities are complex problems and difficult to treat. While many treatment concepts exist for extra-articular deformities there is limited information available for intra-articular or even intracondylar deformities. Selecting the appropriate procedure for the approach, osteotomy, management of bone defects and cartilage damage is crucial. The authors describe the analysis, planning, treatment and clinical outcome for these rare but difficult problems and present step by step details of the surgical technique.

[Trauma sequalae after complex knee ligament injuries]. / Spätzustände nach komplexer Bandverletzung am Kniegelenk.

Complex ligament injuries can compromise a knee joint and residual conditions comprise stiffness (arthrofibrosis), instability, cartilage damage leading to osteoarthritis and bone deformity. Accurate diagnosis must address the direction and extent of the instability, the severity of any cartilage lesion and an analysis of the axis and bone deformity as well as important cofactors. Therapeutic options are adhesiolysis, ligament reconstruction, cartilage regeneration and axis correction. As a consequence patients mostly profit from the procedure but there is never a return to the functional level that existed before injury.

Arguments for an increasing differentiation towards fibrocartilaginous components in midportion Achilles tendinopathy.

PURPOSE: The objective of this study was to investigate the fibrocartilaginous differentiation occurring in midportion Achilles tendinopathy. METHODS: Tendon samples were retrospectively collected from 23 patients, who had undergone surgery for midportion Achilles tendinopathy resistant to conservative treatment. Based on histological scores, the biopts were subdivided into three categories: a light, moderate and severe histopathological stage. Throughout these stages, immunohistochemical staining was performed against biglycan, aggrecan and collagen type II, components characteristic for fibrocartilage. Staining of these components was evaluated using a semi-quantitative scoring method. RESULTS: The immunohistochemical scores of biglycan and aggrecan were statistically significant between the histopathological stages (P < 0.001). The immunohistochemical scores were positively correlated with the increasing histopathological stages [Spearman's correlation coefficient = 0.93 for biglycan and 0.78 for aggrecan (P < 0.001)]. Staining for collagen type II remained negative throughout these stages. CONCLUSION: Immunohistochemical staining of the fibrocartilaginous components biglycan and aggrecan showed a progressive increase, correlated with a further evolved histopathological stage. This observation gave arguments for an increased differentiation towards fibrocartilaginous components at protein level in midportion Achilles tendinopathy.

Influence of rupture patterns of the medial patellofemoral ligament (MPFL) on the outcome after operative treatment of traumatic patellar dislocation.

PURPOSE: Patellar dislocation usually occurs to the lateral side, leading to ruptures of the medial patellofemoral ligament (MPFL) in about 90 % of all cases. Reliable prognostic factors for the stability of the patellofemoral joint after MPFL surgery and satisfaction of the patient have not been established as yet. METHODS: This multicentric study retrospectively included 40 patients with a mean age of 22.4 ± 8.1 years (range 9-48) from 5 German Trauma Departments with first-time traumatic patellar dislocation and operative treatment. Surgery was limited to soft tissue repairs, and a preoperative magnetic resonance imaging (MRI) was performed in all cases. Evaluation of the MRI included sulcus angle, dysplasia of the trochlea, depth and facet asymmetry of the trochlea, Insall-Salvati index, Tibial tuberosity to trochlear groove (TTTG) distance, and rupture patterns of the MPFL. Patients were interrogated after 2 years about recurrent dislocation, satisfaction, and the Kujala score. RESULTS: Trochlea facet asymmetry was significantly lower in patients with redislocation (23.5 ± 18.8) than in patients without redislocation (43.1 ± 16.5, p = 0.03). Patients with a patellar-based rupture were significantly younger (19.5 ± 7.2 years) than patients without patellar-based rupture (25.4 ± 8.1 years, p < 0.02). Patients with femoral-based ruptures were significantly older (25.7 ± 9.2 years) than patients without femoral-based rupture (19.7 ± 6.1 years, p < 0.02), and had a significantly higher TTTG distance (10.2 ± 6.9 vs. 4.5 ± 5.5, p < 0.02). Patients with incomplete ruptures of the MPFL had a significantly lower Insall-Salvati index (1.2 ± 0.2 vs. 1.4 ± 0.2, p = 0.05). The Kujala score in patients with redislocations was significantly lower (81.0 ± 10.5 points) than in patients without redislocation (91.9 ± 9.2 points, p < 0.02). CONCLUSION: Younger patients more often sustain patellar-based ruptures following first-time traumatic patella dislocation, while older patients more often sustain femoral-based ruptures of the MPFL. Incomplete MPFL ruptures are correlated with lower Insall-Salvati indices. Low trochlear facet asymmetry is correlated with higher rates of redislocation. These results may be of relevance for the operative and postoperative treatment in the future. LEVEL OF EVIDENCE: Prognostic study, Level IV.

Anatomic double-bundle ACL reconstruction restricts knee extension in knees with hyperextension.

PURPOSE: Double-bundle ACL reconstruction has been demonstrated to be at least as effective as single-bundle reconstruction in terms of restoring knee rotational and translational stability. Until now, the influence on knees with hyperextension has not been evaluated. It was the purpose of this study to evaluate whether double-bundle ACL reconstruction restricts extension in hyperextendable knees. METHODS: Hamstring tendon reconstructions of 10 human cadaveric knees with the ability of hyperextension (age: 48 ± 14 years) were performed as single bundle (SB) on one side and double bundle (DB) on the other side. A surgical navigation system (BrainLab, Germany) was used to assess the kinematics of each knee at the intact and reconstructed state. A difference with regard to the anterior-to-posterior translation (AP) and rotational stability at 30° of knee flexion, 90° of flexion and the hyperextension capability of each specimen was analysed. RESULTS: The difference in AP translation before and after the reconstruction was not significantly different in 30° and 90° of flexion (n.s). Both single- and double-bundle reconstructions restored the preoperative kinematics at 30° and 90° of knee flexion (n.s). The knee extension was 4° ± 1.8° with the intact ACL and 4° ± 1.7° after reconstruction in the SB group (n.s). The knee extension was 5° of hyperextension ± 1.1° with the intact ACL and 0° ± 0.4° after reconstruction in the DB group; the limitation of the extension was significantly larger in this group (p = 0.013). CONCLUSION: Both single- and double-bundle ACL reconstruction techniques are capable of restoring knee anteroposterior and rotational stability. Double-bundle reconstructions significantly reduce knee extension in knees with hyperextension capability. Care must be taken when using double-bundle techniques in patients with knee hyperextension as this procedure may limit the knee extension after double-bundle ACL reconstruction.

[Implant-free tibial fixations of the posterior cruciate ligament. Development and biomechanical testing]. / Implantatfreie tibiale Fixierung des hinteren Kreuzbandes. Entwicklung und biomechanische Testung.

BACKGROUND: A secure tibial press fit technique in posterior cruciate ligament reconstructions may be a relevant alternative to common techniques because no hardware is necessary. Up to the present point in time no biomechanical data exist for a tibial press fit posterior cruciate ligament (PCL) reconstruction. This study compares the biomechanical properties of hamstring and quadriceps tendon grafts using a press fit technique with those of an interference screw fixation. METHODS: Quadriceps and hamstring tendons of 20 human cadavers (age 49.2±18.5 years) were used. A press fit fixation with a knot in the semitendinosus tendon (K) and a quadriceps tendon bone block graft (Q) were compared to an interference screw fixation (I) in 27 porcine tibiae. In each group, nine constructs were cyclically stretched and then loaded until failure. Maximum load to failure, stiffness and elongation during failure testing and cyclical loading were investigated. RESULTS: The maximum load to failure was 518±157 N (387-650 N) for the K group, 558±119 N (466-650 N) for the I group and 620±102 N (541-699 N) for the Q group. The stiffness was 55±27 N/mm (18-89 N/mm) for the K group, 117±62 N/mm (69-165 N/mm) for the I group and 65±21 N/mm (49-82 N/mm) for the Q group. The stiffness of the I group was significantly larger (ANOVA on ranks, P=0.01). The elongation during cyclical loading was significantly larger for all groups from the 1st to the 5th cycle compared to the elongation in between the 5th and the 20th cycle (P<0.03). CONCLUSION: All techniques exhibited larger elongation during initial loading. Load to failure and stiffness were significantly different between the fixations. The Q fixation showed equal biomechanical properties compared to a pure tendon fixation (I) with an interference screw. The results of group K were inferior. All three investigated fixation techniques exhibit comparable biomechanical properties. Preconditioning of the constructs is critical. Future randomized, clinical trials have to investigate the biological effectiveness of these fixation techniques.

Operative vs conservative treatment of traumatic patellar dislocation: results of a prospective randomized controlled clinical trial.

INTRODUCTION: Patellar dislocation is a common knee injury with mainly lateral dislocations, leading to ruptures of the medial patellofemoral ligament in most of the cases. Even though several prognostic factors for patellofemoral instability have been identified so far, the appropriate therapy for patients with patellar dislocation remains a controversial issue. The purpose of this study was to compare the outcome after conservative or operative treatment in patients after first-time patellar dislocation. PATIENTS AND METHODS: This randomized controlled clinical trial was designed multicentric including patients from six German orthopaedic and trauma departments. Twenty patients with a mean age of 24.6 years with first-time traumatic patella dislocation were included and randomized into either a conservative arm or an operative arm. Plain X-ray images of the knee joint (a.p. and lateral view and tangential view of both patellae) were performed in all cases prior to therapy to exclude osteochondral fragments requiring refixation. An MRI was recommended, but not compulsory. Patients were consulted after 6, 12, and 24 months with a questionnaire including the criteria of the Kujala score, recurrent dislocation, and satisfaction. RESULTS: The mean Kujala score of the conservative vs operative treatment group was 78.6 vs 80.3 after 6 months (p = 0.842), 79.9 vs 88.9 after 12 months (p = 0.165), and 81.3 vs 87.5 after 24 months (p = 0.339). Redislocation rate after 24 months was 37.5 % in the conservative group and 16.7 % in the operative group (p = 0.347). Due to the small number of patients that could be included, no significant difference between the groups could be detected. We see a tendency towards better results after operative treatment. CONCLUSION: Our multicentric prospective randomized controlled trial revealed no significant difference between conservative and operative treatment for patients after first-time traumatic patellar dislocation. However, a tendency towards a better Kujala score and lower redislocation rates for patients with operative treatment was observed. The small number of patients is a limiting factor of the study, leading to results without statistical significance. A meta-analysis including other study's level I data is desirable for the future.

[Biodegradable screw versus a press-fit bone plug fixation for ACL reconstruction: a prospective randomized study]. / Biodegradierbare Schraube vs. einer Press-fit-Verankerung für VKB-Rekonstruktionen : Eine prospektive randomisierte Studie.

BACKGROUND: Press-fit fixation of a tendon graft has been advocated in order to achieve tendon to bone healing. HYPOTHESIS: Fixation of a tendon graft with a porous bone scaffold limits bone tunnel enlargement compared with a biodegradable interference screw fixation. METHODS: Between 2005 and 2006, 20 patients (17 men, 3 women) were enrolled in this study for primary reconstruction of the ACL. Patients were randomized to either obtain graft fixation in the tibial tunnel by means of an interference screw (I) or a press-fit fixation with a porous bone cylinder (P). Three months after surgery, a CT scan of the knee was performed and tunnel enlargement was analysed in the coronal and sagittal planes for the proximal, middle and distal thirds of the tunnel. After 6 months, 1 and 2 years, International Knee Documentation Committee (IKDC), Tegner and Lysholm scores of both groups were compared. RESULTS: The bone tunnel enlargement was 106.9±10.9% for group P and 121.9±9.0% for group I (P<0.02) in the AP plane and 102.8±15.2% vs 121.5±10.1% in the coronal plane (P<0.01). IKDC, Tegner, and Lysholm scores improved in both groups from pre- to postoperative assessment without significant differences between the two groups. There was a trend to higher knee stability in group P after 3 months (0.6±1.4 mm vs 1.81±.5 mm, P=0.08). CONCLUSIONS: Both interference screw and a press-fit fixation lead to a high number of good or very good outcomes after ACL reconstruction. Tibial press-fit fixation decreases the amount of proximal bone tunnel enlargement. Press-fit fixation decreases the amount of proximal bone tunnel enlargement and improves bone to tendon contact.