Type-2 Diabetic (T2D) patients experience up to a 3-fold increase in bone fracture risk[1]. Paradoxically, T2D-patients have a normal or increased bone mineral density when compared to non-diabetic patients. This implies that T2D has a deleterious effect on bone quality, whereby the intrinsic material properties of the bone matrix are altered. Creating clinical challenges as current diagnostic techniques are unable to accurately predict the fracture probability in T2D-patients. To date, the relationship between cyclic fatigue loading, mechanical properties and microdamage accumulation of T2D-bone tissue has not yet been examined and thus our objective is to investigate this relationship. Ethically approved femoral heads were obtained from patients, with (n=8) and without (n=8) T2D. To obtain the mechanical properties of the sample, one core underwent a monotonic compression test to 10% strain, the other core underwent a cyclic compression test at a normalized stress ratio between 0.0035mm/mm and 0.016mm/mm to a maximum strain of 3%. Microdamage was evaluated by staining the tissue with barium sulfate precipitate [2] and conducting microcomputed tomography scanning with a voxel size of 10μm. The monotonically tested T2D-group showed no statistical difference in mechanical properties to the non-T2D-group, even when normalised against BV/TV. There was also no difference in BV/TV. For the cyclic test, the T2D-group had a significantly higher initial modulus (p<0.01) and final modulus (p<0.05). There was no difference in microdamage accumulation. Previous population-level studies have found that T2D-patients have been shown to have an increased fracture risk when compared to non-T2D-patients. This research indicates that T2D does not impair the mechanical properties of trabecular bone from the femoral heads of T2D-patients, suggesting that other mechanisms may be responsible for the increased fracture risk seen in T2D-patients

Abstract. Decellularised porcine superflexor tendon (pSFT) provides an off-the-shelf, cost-efficient option for ACL reconstruction (ACLR). During decellularisation, phosphate buffered saline (PBS) is used for washing out cytotoxic solutes and reagents, maintaining tissue hydration. It has been shown to increase water content in tendon, swelling the tissue reducing mechanical properties. End stage PBS washes in the standard protocol were substituted with alternative solutions to study tissue swelling and its impact on the mechanical behaviour and matrix composition of pSFTs. 25%, 100% Ringers and physiological saline test groups were used (n=6 for all groups). pSFTs were subject to tensile and confined compression testing. Relative hydroxyproline (HYP), glycosaminoglycan (GAG) and denatured collagen content (DNC) were quantified. Modified decellularised tendon groups were compared to tendons decellularised using the standard protocol and native tendons. Specimen dimensions reduced (p=0.004) post-decellularisation only in 25% Ringers group. In all other modified groups, less swelling was apparent but not statistically different from standard group. Only 25% Ringers group had higher linear modulus (p=0.0035) and UTS (p=0.013) compared to standard group. All decellularised groups properties were reduced compared to native pSFTs. Stress relaxation properties showed a significant reduction in decellularised groups compared to native. Compression testing showed no significant differences in peak stress for modified decellularised groups compared to native. A reduction (p=0.036) was observed in standard group. Quantification of GAGs and DNC showed no significant differences between groups. HYP content was higher (p<0.0001) for saline group. A significant reduction in tissue swelling could be related to improved mechanical properties of decellularised pSFTs. Alternative solutions in end stage washes had no significant effect on quantities of matrix components, but altered structure/function could explain the differences in tensile and compressive behaviour, and should be further studied. In all decellularised groups, pSFTs retained suitable mechanical properties for ACLR

With information about a patient's bone mechanical properties orthopaedic operations could be optimised to reduce intra- and post-operative complications. However, there is currently no reliable method of measuring a patient's bone mechanical properties in vivo. We have previously investigated microindentation, using a 1.5mm diameter spherical indenter tip, and found no correlation between these measurements and compression testing measurements. We hypothesised that by using a larger diameter indenter tip we would closer match bone millimetre-scale mechanical properties. 20 bone samples were taken from 20 patients undergoing hip replacement surgery. The samples were machined from the femoral neck calcar cortical bone into 6×3×3mm parallelepiped specimens, aligned with the osteons along the long axis. The samples were micro-computed tomography (CT) scanned to calculate porosity. Microindentation was performed using a 6mm diameter, sapphire, spherical indenter tip. 12 indentations were performed in a grid and the reduced moduli were calculated using the Oliver-Pharr method. Compression testing was then performed to failure and the apparent elastic modulus was calculated for each sample. A moderate correlation was found between the indentation reduced moduli and compression testing elastic moduli (r=0.52, r2=0.275, p=0.018). In addition, a moderate correlation was found between the indentation reduced moduli and CT-measured porosity (r=0.5, r2=0.251, p=0.025) and a strong correlation was found between compression testing moduli and porosity (r=0.75, r2=0.568, p<0.001). Using large-tip spherical microindentation, indentation reduced moduli correlated significantly with compression testing apparent elastic moduli in these 20 cortical bone specimens. Microindentation using a large, spherical indenter tip may predict the mechanical properties of bone at the millimetre length scale and shows promise as a potential future clinical decision aid in surgery

A spine compression fracture is a very common form of fracture in elderly with osteoporosis. Injection of polymethyl methacrylate (PMMA) to fracture sites is a minimally invasive surgical treatment, but PMMA has considerable clinical risks. We develop a novel type thermoplastic injectable bone substitute contains the proprietary composites of synthetic ceramic bone substitute and absorbable thermoplastic polymer. We used thermoplastic biocompatible polymers Polycaproactone (PCL) to encapsulate calcium-based bone substitutes hydroxyapatite (Ca10(PO4)6(OH)2, HA) and tricalcium phosphate (TCP) to form a biodegradable injectable bone composite material. The space occupation ration PCL:HA/TCP is 1:9. After heating process, it can be injected to fracture site by specific instrument and then self-setting to immediate reinforce the vertebral body. The thermoplastic injection bone substitute can obtain good injection properties after being heated by a heater at 90˚C for three minutes, and has good anti-washout property when injected into normal saline at 37˚C. After three minutes, solidification is achieved. Mechanical properties were assessed using the material compression test system and the mechanical support close to the vertebral spongy bone. In vitro cytotoxicity MTT assay (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) was performed and no cell cytotoxicity was observed. In vivo study with three New Zealand rabbits was performed, well bone growth into bone substitute was observed and can maintain good mechanical support after three months implantation. The novel type thermoplastic injection bone substitute can achieve (a) adequate injectability and viscosity without the risk of cement leakage; (b) adequate mechanical strength for immediate reinforcement and prevent adjacent fracture; (c) adequate porosity for new bone ingrowth; (e) biodegradability. It could be developed as a new option for treating vertebral compression fractures

Decreasing the bulk weight without losing the biomechanical properties of commercial pure titanium (Cp-Ti) medical implants is now possible by using Laser Powder Bed Fusion (L-PBF) technology. Gyroid lattice structures that have precious mechanical and biological advantages because of similarity to trabecular bone. The aim of the study was to design and develop L-PBF process parameter optimization for manufacturing gyroid lattice Cp-Ti structures. The cleaning process was then optimized to remove the non-melted powder from the gyroid surface without mechanical loss. Gyroid cubic designs were created with various relative densities by nTopology. L-PBF process parameter optimization was progressed using with Cp-Ti (EOS TiCP Grade2) powder in the EOS M290 machine to achieve parts that have almost full dense and dimensional accuracy. The metallography method was made for density. Dimensional accuracy at gyroid wall thicknesses was investigated between designed and manufactured via stereomicroscope, also mechanical tests were applied with real time experiment and numerical analysis (ANSYS). Mass loss and strut thickness loss were investigated for chemical etching cleaning process. Gyroid parts had 99,5% density. High dimensional accuracy was achieved during L-PBF process parameters optimization. Final L-PBF parameters gave the highest 19% elongation and 427 MPa yield strength values at tensile test. Mechanical properties of gyroid were controlled with changing relative density. A minute chemical etching provided to remove non-melted powders. Compression test results of gyroids at numerical and real-time analysis gave unrelated while deformation behaviors were compatible with each other. Gyroid Cp-Ti osteosynthesis mini plates will be produced with final L-PBF process parameters. MTT cytotoxicity test will be characterized for cell viability. Acknowledgements This project is granted by TUBITAK (120N943). Feza Korkusuz MD is a member of the Turkish Academy of Sciences (TÜBA)

Nuclear factor erythroid 2–related factor 2 (Nrf2) is a crucial transcription factor to maintain cellular redox homeostasis, but is also affecting bone metabolism. As the association between Nrf2 and osteoporosis in elderly females is not fully elucidated, our aim was to shed light on the potential contribution of Nrf2 to the development of age-dependent osteoporosis using a mouse model. Female wild-type (WT, n=18) and Nrf2-knockout (KO, n=12) mice were sacrificed at different ages (12 weeks=young mature adult, and 90 weeks=old), morphological cortical and trabecular properties of femoral bone analyzed by micro-computed tomography (µCT), and compared to histochemistry. Mechanical properties were derived from quasi-static compression tests and digital image correlation (DIC) used to analyze full-field strain distribution. Bone resorbing cells and aromatase expression by osteocytes were evaluated immunohistochemically and empty osteocyte lacunae counted in cortical bone. Wilcoxon rank sum test was used for data comparison and differences considered statistically significant at p<0.05. When compared to old WT mice, old Nrf2-KO mice revealed a significantly reduced trabecular bone mineral density (BMD), cortical thickness (Ct.Th), cortical area (Ct.Ar), and cortical bone fraction (Ct.Ar/Tt.Ar). Surprisingly, these parameters were not different in skeletally mature young adult mice. Metaphyseal trabeculae were thin but present in all old WT mice, while no trabecular bone was detectable in 60% of old KO mice. Occurrence of empty osteocyte lacunae did not differ between both groups, but a significantly higher number of osteoclast-like cells and fewer aromatase-positive osteocytes were found in old KO mice. Furthermore, female Nrf2-KO mice showed an age-dependently reduced fracture resilience when compared to age-matched WT mice. Our results confirmed lower bone quantity and quality as well as an increased number of bone resorbing cells in old female Nrf2-KO mice. Additionally, aromatase expression in osteocytes of old Nrf2-KO mice was compromised, which may indicate a chronic lack of estrogen in bones of old Nrf2-deficient mice. Thus, chronic Nrf2 loss seems to contribute to age-dependent progression of female osteoporosis

Abstract. Objectives. Assess and characterise the suitability of a novel silk reinforced biphasic 3D printed scaffold for osteochondral tissue regeneration. Methods. Biphasic hybrid scaffolds consisted of 3D printed poly(ethylene glycol)-terephthalate-poly(butylene terephthalate)(PEGT/PBT) scaffold frame work (pore size 0.75mm), which has been infilled with a cast and freeze dried porous silk scaffold (5×5×2mm. 3. ), in addition to a seamless silk top layer (1mm). Silk scaffolds alone were used as controls. Both the biphasic and control scaffolds were characterised via uniaxial compression testing (strain rate 0.1mm/min), and the potential biocompatibility of the scaffolds was tested via in vitro culture of seeded bone marrow stromal cells post fabrication. Results. Uniaxial compression testing showed that the biphasic scaffolds (N=4) initially demonstrated similar behaviour on a stress-strain curve to a silk scaffold alone control group (N=6), until a strain of 30% was reached. After 30% strain, load was transitioned from the silk only chondral layer to the 3D printed PEGT/PBT scaffold which resisted further compression and exhibited a significantly greater compressive modulus of 12.6±0.9MPa compared to 0.113±0.01MPa (p<0.001) in the silk scaffold control group. Following 24hours of seeding, no difference was noticed in cell adhesion behaviour under fluorescent microscopy between silk scaffolds and biphasic scaffolds (n=5). Discussion. The use of 3D printing within this novel scaffold provides a solid framework and increases its versatility. The reinforced silk not only provides the secondary Porous structure to the 3D printed scaffold for the bone phase, but also a superficial layer for the cartilage phase. This unique structure has the potential to fill a niche within osteochondral tissue regeneration, especially with the possibility for its use within personalised medicine. Conclusions. These results demonstrate that the novel silk reinforced biphasic 3D printed scaffold is biocompatible and has suitable mechanical properties for osteochondral tissue regeneration. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Abstract. Introduction. Bone grafts are utilised in a range of surgical procedures, from joint replacements to treatment of bone loss resulting from cancer. Decellularised allograft bone is a regenerative, biocompatible and immunologically safe potential source of transplant bone. Objectives. To compare the structural and biomechanical parameters of decellularised and unprocessed (cellular) trabecular bone from the human femoral head (FH) and tibial plateau (TP). Methods. Bone pins were harvested from 10 FHs and 11 TPs (27, 34 respectively). Pins were decellularised (0.1% w/v sodium dodecyl sulphate) or retained as cellular controls. QA testing was carried out to assess protocol efficacy (total DNA and histological analysis). Cellular and decellularised FH (n=7) and TP (n=10) were uCT scanned. Material density (MD); apparent density (BV/TV); trabecular connectivity; trabecular number; trabecular thickness (Tb-t) and trabecular spacing were measured. Pins were then compression tested to determine ultimate compressive stress (UCS), Young's modulus and 0.2% proof stress. Results. Total DNA levels of decellularised bone were below 50 ng.mg. −1. dry weight. Cell nuclei and marrow were largely removed. No significant differences in properties were found between decellularised and cellular bone from either anatomical region (p>0.05, Mann-Whitney). No significant differences in biomechanical properties were found between cellular FH and cellular TP (p>0.05) though significant differences in structural properties were found (MD: TP>FH, p=0.001; BV/TV: FH>TP, p=0.001; and Tb-t: FH>TP, p=0.005). Significant differences were found between decellularised FH and decellularised TP (UCS: FH>TP, p=0.001; Young's modulus: FH>TP, p=0.002; proof stress; FH>TP, p=0.001; MD: TP>FH, p<0.001; BV/TV: FH>TP, p<0.001 and Tb-t: FHT>P p<0.001. Conclusion. Decellularisation did not affect the properties of human trabecular bone. Differences were found between the mechanical and structural properties of decellularised FH and TP which could facilitate stratified bone grafts for different applications. Declaration of Interest. (a) fully declare any financial or other potential conflict of interest

Introduction and Objective. Local cartilage defects in the knee are painful and mostly followed by arthritis. In order to avoid impaired mobility, the osteochondral defect might be bridged by a synthetic compound material: An osteoconductive titanium foam as an anchoring material in the subchondral bone and an infiltrated polymer as gliding material in contact with the surrounding natural cartilage. Materials and Methods. Titanium foam cylinders (Ø38 mm) with porosities ranging from 57% to 77% were produced by powder metallurgy with two different grain sizes of the space holder (fine: 340 ± 110 μm, coarse: 530 ± 160 μm). The sintered titanium foam cylinders were infiltrated with UHMWPE powder on one end and UHMWPE bulk at the other end, at two different temperatures (160 °C, 200 °C), using a pressure of 20 MPa for 15 minutes. Smaller cylinders (Ø16 mm) were retrieved from the compound material by water jet cutting. The infiltration depths were determined by optical microscopy. The anchoring of the UHMWPE was measured by a shear test and the mechanical properties of the titanium foam were verified by a subsequent compression test. The tribological behaviour was investigated in protein containing liquid using fresh cartilage pins (Ø5 mm) sliding against a UHMWPE disc with or without a notch to simulate the gap between the implant and the surrounding cartilage. Friction coefficients were determined in a rotation tribometer and the cartilage wear in a multidirectional six-station tribometer from AMTI (load 10 – 50 N, sliding speed 20 mm/s, 37 °C). Results. UHMWPE could be infiltrated into titanium foam by 1.1 – 1.3 mm with fine pores and by 1.5 – 1.8 mm with coarse pores. The infiltration was neither dependent on the type of UHMWPE (powder or bulk) nor on the temperature. The polymer was so well anchored inside the titanium foam pores that the shear forces for the compounds exceeded the shear strength obtained for a UHMWPE-cylinder. This effect was due to the increased stiffness of the compound plug. Uniaxial compression of the titanium foams after the shear-off of the polymer revealed yield strengths ranging from 50 – 88 MPa for porosities of 62 – 73%. The Ø16 mm samples yielded beyond physiological loads in the knee (≥ 10x body weight) and behaved in a strain hardening and fully ductile manner, reaching deformations of at least 50 % of their initial height without the appearance of macroscopically visible cracks. For smaller plug diameters down to Ø8 mm, however, the lower porosity / higher strength foam should be used to limit elastic deformation of the compound to < 0.1 mm. Pore size did not significantly influence the strength and stiffness values. The elevated coefficient of friction between cartilage and UHMWPE of about 1 was not negatively affected by the presence of the gap. The height loss of the cartilage pin after 1 hour (respectively after 3600 reciproque wear cycles) was 0.2 ± 0.1 mm using a flat disc. For discs with a 1 mm wide V-notch, the wear increased to 0.9 ± 0.3 mm. Conclusions. The tested titanium foams are well suited to act as an anchoring material in the subchondral bone as mechanical properties can be tailored by choosing the adequate porosity and as bone ingrowth has previously been demonstrated for the used pore sizes. UHMWPE is not an ideal gliding partner against cartilage because the friction coefficients of frictions were high. The presence of a V-notched gap was detrimental for cartilage wear. More hydrophilic polymers like PCU should be tested as potential gliding materials

Abstract. Objectives. This study aids the control of remodelling and strain response in bone; providing a quantified map of apparent modulus and strength in the proximal tibia in 3 anatomically relevant directions in terms of apparent density and factor groups. Methods. 7 fresh-frozen cadaveric specimens were quantified computed tomography (qCT) scanned, segmented and packed with 3 layers of 9mm side length cubic cores aligned to anatomical mechanical axes. Cores were removed with printed custom cutting and their densities found from qCT. Cores (n = 195) were quasi-statically compression tested. Modulus was estimated from a load cycle hysteresis loop, between 40% and 20% of yield stress. Sequential testing order in 3 orthogonal directions was randomised. Group differences were identified via an analysis of variance for the factors density, age, gender, testing order, subchondral depth, condyle and sub-meniscal location. Regression models were fit for significant factor sub-groups, predicting properties from density. Results. Axial modulus was 1.5 times greater than the two transverse directions (p<0.001), between which no difference was found. For all test directions, differences were quantified for density and modulus across all subchondral depths (p<0.001). 60% of transverse modulus variation was explained by density within subgroups for each subchondral depth. Medial axial modulus was 1.3 times greater than the lateral side (p = 0.011). Lateral axial modulus halved over a 25mm depth whilst remaining constant for the medial side. Density explained 75% of variation when grouped by subchondral depth and condyle. Yield strength was well predicted across all test directions, with density explaining 81% of axial strength variation and no differences over subchondral depth. Conclusions. The quantification of bone multiaxial modulus based on condyle and subchondral depth has been shown for the first time in a clinically viable protocol using conventional CT. Accounting for spatial variation improves upon literature property prediction models. Declaration of Interest. (b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project

Introduction. In recent years, there has been a growing interest, in many fields of medicine, in the use of bone adhesives that are biodegraded to non-toxic products and resorbed after fulfilling their function in contact with living tissue. Biomechanical properties of newly developed bone glue, such as adhesion to bone and elastic modulus were tested in our study. Material and methods. Newly developed injectable biodegradable “self-setting” bone adhesive prepared from inorganic tricalcium phosphate powder and aqueous solution of organic thermogelling polymers was used for ex-vivo fixing fractured pig femur. Ex-vivo biomechanical tests were performed on 45 fresh pig femurs. Control group consist of 10 healthy bones, tested group was created by 35 bones with artificial fractures in diaphysis – oblique (O) and bending wedge (BW) type of fracture. Tested group were divided to following 4 subgroups (sg); sg1 – O fracture (n=15) glued together with 3 different type of bone adhesives, sg2 BW fracture (n=5) glued together with bone adhesive (n=5); sg3 – BW fracture fixed with locking compression plate (LCP), n=5; sg4 – BW fracture fixed with LCP in combination with bone adhesive. Three-point bending force and shear compression tests were performed on linear electrodynamic test instrument (ElectroPuls E10000, Instron). Femurs from sg1, sg2 and sg4 were tested on Micro-CT before and after biomechanical testing. Results. Shear compression tests in sg1 without amino acids modification showed that it is needed force of 0.5 mPa to recreate fracture, however, modification with amino acids increased glue strength to 3 mPa. Three-point bending force test in sg2 showed reduced force of 250 N to recreate fracture, anyhow in sg4 force needed to initiate the fracture was increased up to 5000 N. Conclusion. Newly developed injectable biodegradable “self- setting” bone adhesive represents new possibility how to fix small bone fragments in comminuted fractures and simultaneous chance how to improve and accelerate bone healing process. Acknowledgement. Project no. AOTEU-R-2016-064 was supported by AOTRAUMA, Switzerland

Introduction. With processing age, meniscus degeneration occurs which is often associated with osteoarthritis. Existing data about the influence of degeneration on the biomechanical properties of the meniscus are still contradictory, or completely unknown regarding the hydraulic permeability. Thus, the aim of this study was to characterise the biomechanical properties and structural composition of the meniscal tissue depending on its degree of degeneration. Methods. Menisci of 24 TKR-patients (≈67.1 yrs.) were harvested and the degeneration of each region (pars anterior PA, pars intermedia PI, pars posterior PP) classified according to Pauli et al. For biomechanical characterisation, confined compression tests (20% strain; velocity: 3%h. 0. /min, relaxation time: 1h) to determine equilibrium modulus (H. A. ) and hydraulic permeability (k) and tensile tests (velocity: 5%l. 0. /min) to determine the tensile modulus were performed. Therefore, cylindrical (Ø= 4.6mm, initial height h. 0. ≈ 2.3mm) and dumbbell-shaped (3.5mm × 1.4mm × 3.5mm) samples were punched out of each region and flattened to achieve parallel surfaces. Additionally, collagen and proteoglycan (PG) content were analysed by calculating the area-under-curve of their specific wavelength ranges (1293–1356cm. −1. and 980–1120cm. −1. , respectively) using infrared (IR) spectroscopy. To identify differences regarding the meniscus regions or its degeneration, a statistically mixed model was used. Results. The compression test showed a significant decrease in H. A. with increasing degeneration (from 78kPa to 55kPa) and from anterior to posterior region (PA: ≈90kPa to PP: ≈70kPa), whereas the hydraulic permeability increased significantly from ≈(1.4 to 3.1)*10. −14. m. 4. N. −1. s. −1. and ≈(1.5 to 3)*10. −14. m. 4. N. −1. s. −1. , respectively. However, the tensile modulus was constant for all regions but showed a decreasing tendency with rising degeneration from ≈(48 to 28)MPa. The collagen content showed a significant decrease with increasing degeneration. The PG content revealed no significant differences regarding the sampling region but a downwards trend with increasing degeneration. Discussion. For the first time, we were able to show a significant increase in the hydraulic permeability with progressive meniscus degeneration while decreasing aggregate modulus. Furthermore, according to a simultaneous downwards tendency in tensile modulus, the collagen content decreased significantly with increasing degeneration. These alterations in biomechanical properties in degenerative meniscal tissue are likely related to an increased water content, also shown e.g. by Pauli et al. In conclusion, our findings may contribute to the understanding of meniscus degeneration and how alterations of meniscal properties might influence the formation of osteoarthritis

Proximal humerus fractures are the third most common fragility fractures with treatment remaining challenging. Mechanical fixation failure rates of locked plating range up to 35%, with 80% of them being related to the screws perforating the glenohumeral joint. Secondary screw perforation is a complex and not yet fully understood process. Biomechanical testing and finite element (FE) analysis are expected to help understand the importance of various risk factors. Validated FE simulations could be used to predict perforation risk. This study aimed to (1) develop an experimental model for single screw perforation in the humeral head and (2) evaluate and compare the ability of bone density measures and FE simulations to predict the experimental findings. Screw perforation was investigated experimentally via quasi-static ramped compression testing of 20 cuboidal bone specimens at 1 mm/min. They were harvested from four fresh-frozen human cadaveric proximal humeri of elderly donors (aged 85 ± 5 years, f/m: 2/2), surrounded with cylindrical embedding and implanted with a single 3.5 mm locking screw (DePuy Synthes, Switzerland) centrally. Specimen-specific linear µFE (ParOSol, ETH Zurich) and nonlinear explicit µFE (Abaqus, SIMULIA, USA) models were generated at 38 µm and 76 µm voxel sizes, respectively, from pre- and post-implantation micro-Computed Tomography (µCT) images (vivaCT40, Scanco Medical, Switzerland). Bone volume (BV) around the screw and in front of the screw tip, and tip-to-joint distance (TJD) were evaluated on the µCT images. The µFE models and BV were used to predict the experimental force at the initial screw loosening and the maximum force until perforation. Initial screw loosening, indicated by the first peak of the load-displacement curve, occurred at a load of 64.7 ± 69.8 N (range: 10.2 – 298.8 N) and was best predicted by the linear µFE (R. 2. = 0.90), followed by BV around the screw (R. 2. = 0.87). Maximum load was 207.6 ± 107.7 N (range: 90.1 – 507.6 N) and the nonlinear µFE provided the best prediction (R. 2. = 0.93), followed by BV in front of the screw tip (R. 2. = 0.89). Further, the nonlinear µFE could better predict screw displacement at maximum force (R. 2. = 0.77) than TJD (R. 2. = 0.70). The predictions of non-linear µFE were quantitatively correct. Our results indicate that while density-based measures strongly correlate with screw perforation force, the predictions by the nonlinear explicit µFE models were even better and, most importantly, quantitatively correct. These models have high potential to be utilized for simulation of more realistic fixations involving multiple screws under various loading cases. Towards clinical applications, future studies should investigate if explicit FE models based on clinically available CT images could provide similar prediction accuracies

The clinical uptake of minimally invasive interventions for intervertebral disc, such as nucleus augmentation, is currently hampered by the lack of robust pre-clinical testing methods that can take into account the large variation in the mechanical behaviour of the tissues. Using computational modelling to develop new interventions could be a way to test scenarios accounting for variation. However, such models need to have been validated for relevant mechanical function, e.g. compressive, torsional or flexional stiffness, and local disc deformations. The aim of this work was to use a novel in-vitro imaging method to assess the validity of computational models of the disc that employed different degrees of sophistication in the anatomical representation of the nucleus. Bovine caudal bone-disc-bone entities (N=6) were dissected and tested in uniaxial compression in a custom-made rig. Forty glass markers were placed on the surface of each disc. The specimens were scanned both with MRI and micro-CT before and during loading. Specimen-specific computational models were built from CT images to replicate the compression test. The anatomy of the nucleus was represented in three ways: assuming a standard diameter ratio, assuming a cylindrical shape with its volume matching that measured from MRI, and deriving the shape directly from MRI. The three types of models were calibrated for force-displacement. The radial displacement of the glass markers were then compared with their experimental displacement derived from CT images. For a similar accuracy in modelling overall force-displacement, the mean error on the surface displacement was 35% for standard ratio nucleus, 38% for image-based cylindrical nucleus, and 32% for MRI-based nucleus geometry. This work shows that, as long as consistency is kept to develop and calibrate image-based computational models, the complexity of the nucleus geometry does not influence the ability of a model to predict surface displacement in the intervertebral disc

Complex pathophysiologies involve different signalling mechanisms, with a multitude of often interconnected potential therapeutic targets. Therefore, there is a need for the development of multi-compartment delivery vehicles for combinatorial and synergistic therapeutic approaches. In this study it was hypothesized that multi-compartment crosslinked collagen type I systems can deliver multiple bioactive agents in a controlled manner in an in vitro model condition of skin fibrosis. Multi-compartment collagen-based systems were made using solutions of dialyzed type I collagen mixed with 10× PBS, after which they were neutralised and crosslinked with 1 and 2.0 mM 4 arm-succinimidyl glutarate ester PEG (4 arm-PEG-SG), respectively, followed by incubation at 37ºC. The systems were characterised through swelling assessment, collagenase degradation assay and compression tests. The release of encapsulated drugs from the hydrogels was studied by ELISA and the effect of the delivered bioactive agents was assessed through imaging and quantification for fibrotic markers in an in vitro model. A pilot study using FITC-dextran proved that the inner compartment was capable of promoting a sustained release over a long period of time (7 days), which was further confirmed with drug release assays using a TNF-α antagonist and recombinant decorin, fitting the intended therapeutic release profile. Protein expression studies showed a decrease of endogenous collagen type I and α-smooth muscle actin expression (p<0.05) indicating amelioration of fibrosis. In summary, this indicates that this system is suitable for dual delivery of multiple bioactive agents, resulting in a controlled release in vitro and illustrating its potential in therapy

Aging has been associated with decreases in muscle strength and bone quality. In elderly patients, paravertebral muscle atrophy is accompanied by vertebral osteoporosis. The purpose of this study was to use paravertebral injection of botulinum toxin-A (BTX) to investigate the effects of paravertebral muscle atrophy on lumbar vertebral bone quality. Forty 16-week-old female SD rats were randomly divided into four groups: (1) a control group (CNT); (2) a resection of erector spinae muscles group (RESM); (3) a botulinum toxin-A group (BTX) that was treated with local injection of 5U BTX into the paravertebral muscles bilaterally; and (4) a positive control group (OVX) that underwent bilateral ovariectomy. At 3 months post-surgery the lumbar vertebrae (L3 – L6) were collected. The BMDs of the RESM and BTX groups were significantly lower than that of the CNT group (P < 0.01). Micro-CT scans showed that rats in the three experimental groups had fewer trabeculae and trabecular connections than rats in the CNT group. The bone loss trend of the trabecular networks was most obvious in the OVX rats. Vertebral compression testing revealed that the three experimental groups had significantly lower maximum load, energy absorption, maximum stress, and elastic modulus values than the CNT group (P < 0.01), and these parameters were lowest in the OVX group (P < 0.05). Our results demonstrate that the new paravertebral muscle atrophy model using local BTX injection causes sufficient muscle atrophy and dysfunction to result in local lumbar vertebral bone loss and quality deterioration

Scaffold-based bone tissue engineering holds great promise for the future of osseous defects therapies. Prepare the suitable scaffold properties are physiochemical modifications in terms of porosity, mechanical strength, cell adhesion, biocompatibility, cell proliferation, mineralization and osteogenic differentiation are required. We produce various bone tissue scaffolds with different techniques such as lyophilization, 3D printing and electrospinning. We wish to overview all the different novel scaffold methods and materials. To improve scaffolds poor mechanical properties, while preserving the porous structure, it is possible to coat the scaffold with synthetic or natural polymers. An increasing interest in developing materials in bone tissue engineering is directed to the organic/inorganic composites that mimic natural bone. Specifically, bone tissue is a composite of an organic and inorganic matrix. Using PLLA, loofah, chitin and cellulose biomaterials we produced bone tissue scaffold with lyophilization technique. Also, using fish scale powder and wet electrospun Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) a sponge structure had created. Using MRI image data and 3D printer method, a bone tissue scaffold is created by PLA filament. Their mechanical properties had analysed with compression tests and their biocompatibilities had investigated. In order to provide novel strategies for future treatment of bone tumours, the properties of the scaffold, including its in vitro extended-release properties, the inhibition effects of chemotherapeutic agent on the bone tumours and its bone repair capacities were investigated in vitro by using MG63 cells. To develop chemotherapeutic agent-encapsulated poly(lactic-co-glycolic acid) (PLGA) nanoparticles in a porous nano-hydroxyapatite scaffold we aimed to use double emulsion method

The complex structural arrangement of bone gives rise to anisotropic, rate-dependent failure behaviour, which varies significantly depending on tissue composition and architecture. This presents significant challenges in the development of orthopaedic surgical cutting instruments, which are required to generate sufficient forces to penetrate bone tissue, while minimizing the risk of thermal and mechanical damage to the surrounding environment. Currently, instrument designers rely heavily on empirical-based strategies to understand tool-bone interactions, with significant amounts of prototyping and validation experiments required throughout the design process. The aim of this study is to develop an experimentally-validated predictive computational model of orthopaedic cutting processes in three dimensions to understand the role of various cutting parameters on cutting forces and chip formation. An experimental model of orthogonal cutting was developed, whereby an adaptable cutting tool fixture driven by a servo-hydraulic uniaxial test machine was used to carry out high-rate cutting tests on Sawbone® trabecular bone analogues. A three-dimensional computational model was also developed using Abaqus/Explicit. The constitutive model describing material behaviour considers strain-rate and pressure-dependant yield behaviour using a Drucker-Prager elastic-plastic damage model, with Strain-hardening and rate-dependent model constants determined through dynamic uniaxial high-strain rate compression tests of material cubes. An excellent correlation between experimental and computational models was found, with the computational model accurately predicting tool cutting forces and chip development ahead of the tool during the cutting process. It was identifying that lower tool rake-angles resulted in the formation of larger discontinuous chips and higher cutting forces, while higher rake angles tended to lead to more continuous chip formation and lower cutting forces

Osteoarthritis (OA) affects over 8.75 million people in the UK creating the need for early stage interventions. Osteochondral (OC) grafting has been used to repair full thickness lesions but the efficacy of this therapy is questionable. As a first step in developing a testing framework able to predict the potential and suitability of OC grafts for repair, here, we present experimental data to be used in informing boundary conditions, input parameters and testing sequences for developing and verifying an FE model of the interaction of OC grafts and surrounding host tissue. Ten OC cylindrical grafts (height: 10mm; diameter: n=5–6.5mm; n=5–8.5mm) were harvested from adult porcine femurs (60–70kg). Unconfined compression tests were conducted using an Instron3365 with a 500N load cell and a BioBath filled with PBS at 37ºC. The OC grafts (prior to separation of cartilage and bone) and cartilage underwent four 5% strain (of cartilage layer) steps with displacement rate of 0.005mm/sec, each followed by a 45-minute relaxation. Final strain was 20%. Bone underwent a single displacement of 20% strain of bone at same displacement rate. Young's moduli ranged from 6.2–42.0MPa, 0.7–3.9MPa, 46.8–123.7MPa for OC graft, cartilage and bone, respectively. The coefficient of variance between OC Grafts, cartilage and bone was 70.6%, 71.8%, and 25.2%, respectively. Dispersion between samples was high. This may be due to intrinsic tissue variability but also due to the testing protocol: for cartilage in particular, the load was at the low end of the load cell capacity and the sample aspect ratio was poor for compressive testing. This work provides insight in understanding the effect of individual patient's and/or individual grafts used during osteochondral grafting. The results compel the need to accurately model these tissues when developing specimen-specific FE models for OC grafting

Background. Skeletal stem cells can be combined with human allograft, and impacted to produce a mechanically stable living bone composite. This strategy has been used for the treatment of femoral head avascular necrosis, and has been translated to four patients, of which three remain asymptomatic at up to three year follow-up. In one patient collapse occurred in both hips due to widely distributed and advanced AVN disease, necessitating bilateral hip arthroplasty. However this has provided the opportunity to retrieve the femoral heads and analyse human tissue engineered bone. Aims. Analysis of retrieved human tissue-engineered bone in conjunction with clinical follow-up of this translational case series. Methods. A parallel in vitro culture of the implanted cell-graft constructs was set up at the time of surgery, with serial cell viability stains performed up to six weeks. Patient follow-up was by serial clinical and radiological examination. Tissue engineered bone from the two retrieved femoral heads was analysed histologically by Alcian blue & Sirius red stain and bi-refringence, by micro computed tomography (microCT) for both bone density and morphology, and by compression testing for mechanical strength. Normal trabecular and cortical bone from the femoral heads was used as controls. Results. Parallel in vitro analysis demonstrated sustained cell growth and viability on the allograft. Histologically, the retrieved tissue engineered specimens demonstrated a mature trabecular micro-architecture and organization identical to normal trabecular bone. MicroCT revealed trabecular morphology within the tissue-engineered bone, with bone density of 1400 Grey scale units (compared to 1200 for natural trabecular bone and 1800 for cortical bone). Axial compression testing showed no difference in strength between engineered and trabecular bone. Conclusions. Widespread residual necrosis in the femoral heads of one patient resulted in collapse requiring hip arthroplasty, but analysis of the tissue engineered bone sections has demonstrated the translational potential of a living bone composite to restore both the biological and mechanical characteristics of bone defects. Clinical follow-up shows this to be an effective new treatment for focal early stage avascular necrosis of the femoral head, and this unique retrieval analysis data confirms the potential of cell-based strategies for clinical treatment of bone defects