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Orthopaedic Proceedings
Vol. 103-B, Issue SUPP_4 | Pages 69 - 69
1 Mar 2021
Sahm F Grote VF Detsch R Kreller T Boccaccini A Bader R Jonitz-Heincke A
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Several electrical fields are known to be present in bone tissue as originally described by Fukada and Yasuda in the year 1957. Intrinsic voltages can derive from bone deformation and reversely lead to mechanical modifications, called the piezoelectric effect. This effect is used in the clinic for the treatment of bone defects by applying electric and magnetic stimulation directly to the bone supplied with an implant such as the electroinductive screw system. Through this system a sinusoidal alternating voltage with a maximum of 700 mV can be applied which leads to an electric field of 5–70 V/m in the surrounding bone. This approach is established for bone healing therapies. Despite the established clinical application of electrical stimulation in bone, the fundamental processes acting during this stimulation are still poorly understood. A better understanding of the influence of electric fields on cells involved in bone formation is important to improve therapy and clinical success.

To study the impact of electrical fields on bone cells in vitro, Ti6Al4V electrodes were designed according to the pattern of the ASNIS III s screw for a 6-well system. Osteoblasts were seeded on collagen coated coverslip and placed centred on the bottom of each well. During four weeks the cells were stimulated 3×45 min/d and metabolic and alkaline phosphatase (ALP) activity as well as gene expression of cells were analysed. Furthermore, supernatants were collected and proteins typical for bone remodelling were examined.

The electrical stimulation did not exert a significant influence on the metabolic activity and the ALP production in cells over time using these settings. Gene expression of BSP and ALP was upregulated after the first 3 days whereas OPG was increased in the second half after 14 days of electrical stimulation. Moreover, the concentration of the released proteins OPG, IL-6, DKK-1 and OPN increased when cells were cultivated under electrical stimulation. However, no changes could be seen for essential markers, like RANKL, Leptin, BMP-2, IL-1beta and TNF-alpha.

Therefore, further studies will be done with osteoblasts and osteoclasts to study bone remodelling processes under the influence of electrical fields more in detail. This study was supported by the German Research Foundation (DFG) JO 1483/1-1.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_16 | Pages 83 - 83
1 Nov 2018
Paulus AC Ebinger K Haßelt S Kretzer JP Bader R Utzschneider S
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The biological reaction in metallosis and pseudotumor generation after metal on metal total hip arthroplasty or corroding metal implants remains unsettled. Clinically, still lethal cases appear with massive bone loss and metal ions are suspected to be responsible for this inflammatory reaction, solid metal wear particles instead are usually not observed in the common literature. The aim of this study was to compare the biological reactions of metal ions and metal wear particles in a murine in vivo model. Metal ions (CoCr), metal particles (CoCr), polyethylene particles (UHMWPE) and phosphate buffered saline (PBS) were injected into the left knee joint of female BALB/c mice. 7 days after injection, the microcirculation was observed using intravital fluorescence microscopy, followed by euthanasia of the animals. After the assessment of the knee diameter, the knees underwent histological evaluations of the synovial layer. Throughout all recorded data, CoCr particles caused higher inflammatory reactions compared to metal ions and UHMWPE particles. The mice treated with the solid particles showed enlarged knee diameters, more intensive leukocyte–endothelial cell interactions and an elevated functional capillary density. Pseudotumor-like tissue formations in the synovial layer of the mice were only seen after the exposition to solid CoCr particles. Even if the focus of several national guidelines concerning metallosis and pseudotumor generation is on metal ions, the present data reveal that solid CoCr particles have the strongest inflammatory activity compared with metal ions and UHMWPE particles in vivo.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_4 | Pages 4 - 4
1 Apr 2018
Ziebart J Sellmann T Porath K Delenda B van Rienen U Bader R Köhling R
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Introduction

Migration of bone cells and precursor cells to the site of a bone defect can accelerate bone regeneration. Therefore, guidance of these cells by direct current (DC) is an interesting approach to improve implant ingrowth or fracture healing. To allow a better understanding of DC-induced directed migration, a specific stimulation chamber was established and the influence of DC on calcium channel expression in osteoblasts was investigated.

Methods

Human osteoblasts were isolated from femoral heads of patients undergoing total hip arthroplasty after patient”s consent. The study was approved by the local ethical committee (AZ: 2010–10). Differentiation into osteoblasts was ensured by cultivation in standard cell culture medium enriched with β-glycerophosphate, ascorbic acid and dexamethasone. 2×103 osteoblasts were seeded into custom-made chambers for DC field application. After 12 h DC was applied to chambers via Ag/AgCl electrodes set into separate reservoirs coupled to cell culture area by 2% agarose bridges in order to prevent cytotoxic impact of electrochemical reactions proceeding at the electrodes. Electric fields ranging from 150 to 450 V/m were applied to cells for 7 h. Several cell images were taken over time and used for evaluation of migration direction and speed with ImageJ software. Subsequently, cells were lysed in Trizol for RNA isolation and semiquantitative real-time polymerase chain reaction of voltage-gated calcium channels Cav1.4 and Cav3.2 as well as stretch-activated magnesium and calcium channel TRPM7 was performed.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_4 | Pages 48 - 48
1 Apr 2018
Kebbach M Grawe R Geier A Winter E Kluess D Woernle C Bader R
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Introduction

Despite decades of clinical research in artificial joints and underlying failure mechanisms, systematical and reproducible identification of reasons for complications in total knee replacements (TKR) remains difficult. Due to the complex dynamic interaction of implant system and biological situs, malfunction eventually leading to failure is multifactorial and remains not fully understood. The aim of present study was to evaluate different TKR designs and positions with regard to joint kinematics and stability under dynamic conditions by using a robot-based hardware-in-the-loop (HiL) setup.

Material & methods

An industrial 6-axis robot with 6-axis force-torque sensor mounted into its end-effector moved and loaded real, commercially available TKR (bicondylar, cruciate-retaining) that were in virtual interaction with a subject-specific computational multibody model representing the anatomical situs of the knee joint while performing passive seated deep knee flexion. The subject-specific musculoskeletal multibody model (MMB) included rigid bones of the lower right extremity. Bone and cartilage geometries were reconstructed from MRT/ CT data sets preserving anatomical landmarks and allowing for the calculation of inertial properties. M. quadriceps femoris was modeled as single passive tensile force elements. Knee ligaments were modelled as elastic spring elements with a nonlinear force-displacement characteristic. Providing the flexion angle, the robot moved and loaded the mounted femoral implant component with respect to the tibial component while being in continuous interaction with the MMB. Several influencing parameters like implant position (internal/external rotation, varus/valgus alignment) and design (fixed vs. mobile bearing, tibia-insert height) as well as ligament insufficiency and joint loading on joint kinematics and stability was systematically analysed.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_4 | Pages 42 - 42
1 Apr 2018
Gabler C Gierschner S Lindner T Tischer T Bader R
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The biomechanical evaluation of tendon repair with collagen-based scaffolds in rat model is a common method to determine the functional outcome of the tested material. We introduced a magnetic resonance imaging (MRI) approach to verify the biomechanical test data. In present study different collagen scaffolds for tendon repair were examined.

Two collagen test materials: based on bovine stabilized collagen, chemically cross-linked with oriented collagenous fibres (material 1) and based on porcine dermal extracellular matrix, with no cross-linking (material 2) were compared. The animal study was approved by the local review board. Surgery was performed on male Sprague-Dawley rats with a body weight of 400 ± 19 g. Each rat underwent a 5 mm transection of the right Achilles tendon. The M. plantaris tendon was removed. The remaining tendon ends were re-joined with a 5 mm scaffold of either the material 1 or 2. Each scaffold material was sutured into place with two single stiches (Vicryl 4–0, Ethicon) each end. A total of 16 rats (n= 8 each group) were observed for 28 days follow up. The animals were sacrificed and hind limbs were transected proximal to the knee joint. MRI was performed using a 7 Tesla scanner (BioSpec 70/30, Bruker). T2-weighted TurboRARE sequences with an in-plane resolution of 0.12 mm and a slice thickness of 0.7 mm were analysed. All soft and hard tissues were removed from the Achilles tendon-calcaneus-foot complex before biomechanical testing. Subsequently, the specimens were fixed in a materials testing machine (Z1.0, Zwick, Ulm, Germany) for tensile testing. All tendons were preloaded with 1 N and subsequently stretched at a rate of 1 mm/s until complete failure was observed. Non-operated tendons were used as a control (n=4).

After 28 postoperative days, MRI demonstrated that four scaffolds (material 1: n=2, material 2: n=2) were slightly dislocated in the proximal part of hind limb. In total five failures of reconstruction could be detected in the tendon repairs (material 1: n=3, material 2: n=2). Tendons augmented with the bovine material 1 showed a maximum tensile load of 57.9 ± 17.9 N and tendons with porcine scaffold material 2 of 63.1 ± 19.5 N. The native tendons demonstrated only slightly higher loads of 76.6 ± 11.6 N. Maximum failure load of the tendon-scaffold construct in both groups did not differ significantly (p < 0.05). Stiffness of the tendons treated with the bovine scaffold (9.9 ± 3.6 N/mm) and with the porcine scaffold (10.7 ± 2.7 N/mm) showed no differences. Stiffness of the native healthy tendon of the contralateral site was significantly higher (20.2 ± 6.6 N/mm, p < 0.05). No differences in the mechanical properties between samples of both scaffold groups could be detected, regardless of whether the repaired tendon defect has failed or the scaffold has been dislocated.

The results show that MRI is important as an auxiliary tool to verify the biomechanical outcome of tendon repair in animal models.


Orthopaedic Proceedings
Vol. 100-B, Issue SUPP_4 | Pages 45 - 45
1 Apr 2018
Markhoff J Weinmann M Schulze C Nebe B Bader R
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Nowadays, biomaterials can be used to maintain or replace several functions of the human body being constricted or lost due to tumors, fractures, injuries as well as chronic diseases, infections or simply aging. Titanium and its alloys, i.e. Ti6Al4V are the most common materials (70 to 80%) used for structural orthopedic implants due to their unique combination of good mechanical properties, corrosion resistance and biocompatibility. Addition of β-stabilizers, e. g. niobium (Nb), can improve the mechanical properties of such titanium alloys further, simultaneously offering excellent biocompatibility. Previous studies concerning biocompatibility analyses with niobium and especially Ti-42Nb specimens are rarely described; none for niobium and Ti-42Nb powders examining human cell viability, collagen and interleukin synthesis. In this in vitro study, human osteoblasts were cultured on different roughened niobium specimens (Nb Amperit, Nb Ampertec), Nb sheets and spherical Ti-42Nb (sintered and 3D-printed by selective laser melting, SLM) and compared with forged Ti6Al4V specimens. Furthermore, human osteoblasts were incubated with particulates of the Nb and Ti-42Nb specimens in three particle concentrations over four and seven days to imitate influence of wear debris against the background of osteolysis and aseptic implant loosening. Thereby, the specimens with the roughest surfaces, i.e. Ti-42Nb and Nb Ampertec, revealed excellent and similar results concerning cell viability (WST-1 test, live-dead staining) and collagen-I synthesis superior to forged Ti6Al4V. Examinations with particulate debris disclosed a significant dose-dependent influence of all powders with Nb Ampertec showing the highest decrease of cell viability and collagen-I synthesis. Furthermore, interleukin expression was only slightly increased for all powders. In summary, from a cell-biological point of view Nb Ampertec (sintered Nb) and Ti-42Nb materials seem to be superior alternatives for medical applications compared to common materials like forged Ti6Al4V.