Nuclear factor erythroid 2-related factor 2 (Nrf2) deficiency causes age-dependent progression of female osteoporosis.

BACKGROUND: Nuclear factor erythroid 2-related factor 2 (Nrf2) is a crucial transcription factor for cellular redox homeostasis. The association of Nrf2 with elderly female osteoporotic has yet to be fully described. The aim was to elucidate a potential age-dependent Nrf2 contribution to female osteoporosis in mice. METHODS: Eighteen female wild type (WT) and 16 Nrf2-knockout (KO) mice were sacrificed at different ages (12 weeks = young mature adult and 90 weeks = old) to analyze their femurs. The morphological properties (trabecular and cortical) were evaluated by micro-computed tomography (µCT) and compared to gold standard histochemistry analysis. The quasi-static compression tests were performed to calculate the mechanical properties of bones. Additionally, the population of bone resorbing cells and aromatase expression by osteocytes was immunohistochemically evaluated and empty osteocyte lacunae was counted in cortical bone. RESULTS: Old Nrf2-KO mice revealed a significantly reduced trabecular bone mineral density (BMD), cortical thickness, cortical area, and bone fraction compared to old WT mice, regardless of no significant difference in skeletally mature young adult mice between WT and KO. Specifically, while all old WT mice showed thin metaphyseal trabeculae, trabecular bone was completely absent in 60% of old KO mice. Additionally, old KO mice showed significantly more osteoclast-like cells and fewer aromatase-positive osteocytes than WT mice, whereas the occurrence of empty osteocyte lacunae did not differ between both groups. Nrf2-KO mice further showed an age-dependently reduced fracture resilience compared to age-matched WT mice. CONCLUSION: Our results suggest that chronic Nrf2 loss can lead to age-dependent progression of female osteoporosis.

High-Precision 3D Printing of Microporous Cochlear Implants for Personalized Local Drug Delivery.

Hearing loss is a highly prevalent multifactorial disorder affecting 20% of the global population. Current treatments using the systemic administration of drugs are therapeutically ineffective due to the anatomy of the cochlea and the existing blood-labyrinth barrier. Local drug delivery systems can ensure therapeutic drug concentrations locally while preventing adverse effects caused by high dosages of systemically administered drugs. Here, we aimed to design, fabricate, and characterize a local drug delivery system for the human cochlea. The design was relevant to the size of the human ear, included two different shapes, and incorporated two different microporous structures acting as reservoirs for drug loading and release. The four cochlear implant designs were printed using the two-photon polymerization (2PP) technique and the IP-Q photoresist. The optimized 2PP process enabled the fabrication of the cochlear implants with great reproducibility and shape fidelity. Rectangular and cylindrical implants featuring cylindrical and tapered tips, respectively, were successfully printed. Their outer dimensions were 0.6 × 0.6 × 2.4 mm3 (L × W × H). They incorporated internal porous networks that were printed with high accuracy, yielding pore sizes of 17.88 ± 0.95 µm and 58.15 ± 1.62 µm for the designed values of 20 µm and 60 µm, respectively. The average surface roughness was 1.67 ± 0.24 µm, and the water contact angle was 72.3 ± 3.0°. A high degree of polymerization (~90%) of the IP-Q was identified after printing, and the printed material was cytocompatible with murine macrophages. The cochlear implants designed and 3D printed in this study, featuring relevant sizes for the human ear and tunable internal microporosity, represent a novel approach for personalized treatment of hearing loss through local drug delivery.

In Vivo Prevention of Implant-Associated Infections Caused by Antibiotic-Resistant Bacteria through Biofunctionalization of Additively Manufactured Porous Titanium.

Additively manufactured (AM) porous titanium implants may have an increased risk of implant-associated infection (IAI) due to their huge internal surfaces. However, the same surface, when biofunctionalized, can be used to prevent IAI. Here, we used a rat implant infection model to evaluate the biocompatibility and infection prevention performance of AM porous titanium against bioluminescent methicillin-resistant Staphylococcus aureus (MRSA). The specimens were biofunctionalized with Ag nanoparticles (NPs) using plasma electrolytic oxidation (PEO). Infection was initiated using either intramedullary injection in vivo or with in vitro inoculation of the implant prior to implantation. Nontreated (NT) implants were compared with PEO-treated implants with Ag NPs (PT-Ag), without Ag NPs (PT) and infection without an implant. After 7 days, the bacterial load and bone morphological changes were evaluated. When infection was initiated through in vivo injection, the presence of the implant did not enhance the infection, indicating that this technique may not assess the prevention but rather the treatment of IAIs. Following in vitro inoculation, the bacterial load on the implant and in the peri-implant bony tissue was reduced by over 90% for the PT-Ag implants compared to the PT and NT implants. All infected groups had enhanced osteomyelitis scores compared to the noninfected controls.

The effects of lower extremity muscle fatigue on the vertical ground reaction force: a meta-analysis.

There is currently no consensus in the literature on whether the magnitude of the ground reaction force or loading rate decreases or increases with muscle fatigue. In this article, the effects of lower extremity muscle fatigue on the magnitude of the ground reaction force and loading rate during running and drop landing are examined. Through a systematic search of the literature, 24 articles are identified that satisfy the inclusion criteria and study the relationship between fatigue and the ground reaction force variables during running, single-leg drop landing, and bilateral drop landing. The articles are categorized based on the type of locomotion they study. To determine whether or not the ground reaction force peaks/loading rate are markedly affected by fatigue, meta-analyses are conducted both separately for each type of locomotion and for an aggregate of all studies. The results of the meta-analyses show that the ground reaction force active peak significantly decreases for the following study groups: aggregate of all included studies, studies of drop landing (both single-leg and bilateral), and studies of bilateral landing only. The ground reaction force active peak did not significantly change in running and single-leg drop landing. The effects of muscle fatigue on the magnitude of the ground reaction force impact peak and loading rate was always insignificant.

Additively manufactured biodegradable porous metals.

Partially due to the unavailability of ideal bone substitutes, the treatment of large bony defects remains one of the most important challenges of orthopedic surgery. Additively manufactured (AM) biodegradable porous metals that have emerged since 2018 provide unprecedented opportunities for fulfilling the requirements of an ideal bone implant. First, the multi-scale geometry of these implants can be customized to mimic the human bone in terms of both micro-architecture and mechanical properties. Second, a porous structure with interconnected pores possesses a large surface area, which is favorable for the adhesion and proliferation of cells and, thus, bony ingrowth. Finally, the freeform geometrical design of such biomaterials could be exploited to adjust their biodegradation behavior so as to maintain the structural integrity of the implant during the healing process while ensuring that the implant disappears afterwards, paving the way for full bone regeneration. While the AM biodegradable porous metals that have been studied so far have shown many unique properties as compared to their solid counterparts, the unprecedented degree of flexibility in their geometrical design has not yet been fully exploited to optimize their properties and performance. In order to develop the ideal bone implants, it is important to take advantage of the full potential of AM biodegradable porous metals through detailed and systematic study on their biodegradation behavior, mechanical properties, biocompatibility, and bone regeneration performance. This review paper presents the state of the art in AM biodegradable porous metals and is focused on the effects of material type, processing, geometrical design, and post-AM treatments on the mechanical properties, biodegradation behavior, in vitro biocompatibility, and in vivo bone regeneration performance of AM porous Mg, Fe, and Zn as well as their alloys. We also identify a number of knowledge gaps and the challenges encountered in adopting AM biodegradable porous metals for orthopedic applications and suggest some promising areas for future research.

Functional evaluation of a non-assembly 3D-printed hand prosthesis.

In developing countries, the access of amputees to prosthetic devices is very limited. In a way to increase accessibility of prosthetic hands, we have recently developed a new approach for the design and 3D printing of non-assembly active hand prostheses using inexpensive 3D printers working on the basis of material extrusion technology. This article describes the design of our novel 3D-printed hand prosthesis and also shows the mechanical and functional evaluation in view of its future use in developing countries. We have fabricated a hand prosthesis using 3D printing technology and a non-assembly design approach that reaches certain level of functionality. The mechanical resistance of critical parts, the mechanical performance, and the functionality of a non-assembly 3D-printed hand prosthesis were assessed. The mechanical configuration used in the hand prosthesis is able to withstand typical actuation forces delivered by prosthetic users. Moreover, the activation forces and the energy required for a closing cycle are considerably lower as compared to other body-powered prostheses. The non-assembly design achieved a comparable level of functionality with respect to other body-powered alternatives. We consider this prosthetic hand a valuable option for people with arm defects in developing countries.

A model-based parametric study of impact force during running.

This paper deals with the impact force during foot-ground impact activities such as the running. A previously developed model is used for this study. The model is a lumped-parameter one consisting of four masses connected to each other via linear springs and viscous dampers. A shoe-specific nonlinear function is used for representation of the ground reaction force. The authors have previously showed that the previous version of the model as well as its simulation is incorrect. This paper slightly modifies the previous model so as it is able to produce results in agreement with the experiments. Then, the modified model is simulated for two typical shoe types. A parametric study is also conducted. The parametric study concerns with the effects of masses, mass ratios, stiffness constants, and damping coefficients on the dynamics of the impact. It is shown that the impact forces increase as the rigid and wobbling masses increase. However, the increase in the impact forces is not the same for all the masses. It is found that the impact force increases as the touchdown velocities increase. Simulations imply that the variations of the damping coefficients result in larger variations of the impact force compared to the stiffness. The effect of the variation of gravity on the simulated impact force is also explored. It is concluded that both the first and the second peaks of the impact force are increased with gravity. An in-depth discussion is included to compare results of the current paper with results of other investigators.

Analytical relationships for prediction of the mechanical properties of additively manufactured porous biomaterials.

Recent developments in additive manufacturing techniques have motivated an increasing number of researchers to study regular porous biomaterials that are based on repeating unit cells. The physical and mechanical properties of such porous biomaterials have therefore received increasing attention during recent years. One of the areas that have revived is analytical study of the mechanical behavior of regular porous biomaterials with the aim of deriving analytical relationships that could predict the relative density and mechanical properties of porous biomaterials, given the design and dimensions of their repeating unit cells. In this article, we review the analytical relationships that have been presented in the literature for predicting the relative density, elastic modulus, Poisson's ratio, yield stress, and buckling limit of regular porous structures based on various types of unit cells. The reviewed analytical relationships are used to compare the mechanical properties of porous biomaterials based on different types of unit cells. The major areas where the analytical relationships have improved during the recent years are discussed and suggestions are made for future research directions. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3164-3174, 2016.

Mechanical Properties of Additively Manufactured Thick Honeycombs.

Honeycombs resemble the structure of a number of natural and biological materials such as cancellous bone, wood, and cork. Thick honeycomb could be also used for energy absorption applications. Moreover, studying the mechanical behavior of honeycombs under in-plane loading could help understanding the mechanical behavior of more complex 3D tessellated structures such as porous biomaterials. In this paper, we study the mechanical behavior of thick honeycombs made using additive manufacturing techniques that allow for fabrication of honeycombs with arbitrary and precisely controlled thickness. Thick honeycombs with different wall thicknesses were produced from polylactic acid (PLA) using fused deposition modelling, i.e., an additive manufacturing technique. The samples were mechanically tested in-plane under compression to determine their mechanical properties. We also obtained exact analytical solutions for the stiffness matrix of thick hexagonal honeycombs using both Euler-Bernoulli and Timoshenko beam theories. The stiffness matrix was then used to derive analytical relationships that describe the elastic modulus, yield stress, and Poisson's ratio of thick honeycombs. Finite element models were also built for computational analysis of the mechanical behavior of thick honeycombs under compression. The mechanical properties obtained using our analytical relationships were compared with experimental observations and computational results as well as with analytical solutions available in the literature. It was found that the analytical solutions presented here are in good agreement with experimental and computational results even for very thick honeycombs, whereas the analytical solutions available in the literature show a large deviation from experimental observation, computational results, and our analytical solutions.

Revival of pure titanium for dynamically loaded porous implants using additive manufacturing.

Additive manufacturing techniques are getting more and more established as reliable methods for producing porous metal implants thanks to the almost full geometrical and mechanical control of the designed porous biomaterial. Today, Ti6Al4V ELI is still the most widely used material for porous implants, and none or little interest goes to pure titanium for use in orthopedic or load-bearing implants. Given the special mechanical behavior of cellular structures and the material properties inherent to the additive manufacturing of metals, the aim of this study is to investigate the properties of selective laser melted pure unalloyed titanium porous structures. Therefore, the static and dynamic compressive properties of pure titanium structures are determined and compared to previously reported results for identical structures made from Ti6Al4V ELI and tantalum. The results show that porous Ti6Al4V ELI still remains the strongest material for statically loaded applications, whereas pure titanium has a mechanical behavior similar to tantalum and is the material of choice for cyclically loaded porous implants. These findings are considered to be important for future implant developments since it announces a potential revival of the use of pure titanium for additively manufactured porous implants.

Additively manufactured porous tantalum implants.

The medical device industry's interest in open porous, metallic biomaterials has increased in response to additive manufacturing techniques enabling the production of complex shapes that cannot be produced with conventional techniques. Tantalum is an important metal for medical devices because of its good biocompatibility. In this study selective laser melting technology was used for the first time to manufacture highly porous pure tantalum implants with fully interconnected open pores. The architecture of the porous structure in combination with the material properties of tantalum result in mechanical properties close to those of human bone and allow for bone ingrowth. The bone regeneration performance of the porous tantalum was evaluated in vivo using an orthotopic load-bearing bone defect model in the rat femur. After 12 weeks, substantial bone ingrowth, good quality of the regenerated bone and a strong, functional implant-bone interface connection were observed. Compared to identical porous Ti-6Al-4V structures, laser-melted tantalum shows excellent osteoconductive properties, has a higher normalized fatigue strength and allows for more plastic deformation due to its high ductility. It is therefore concluded that this is a first step towards a new generation of open porous tantalum implants manufactured using selective laser melting.

Relationship between in vitro apatite-forming ability measured using simulated body fluid and in vivo bioactivity of biomaterials.

In a large number of studies, it has been assumed that the in vitro apatite-forming ability measured by simulated body fluid (SBF) test is a predictor of in vivo bioactivity. Several researchers have argued in favor and against this assumption; but the actual experimental evidence is not yet fully examined. The purpose of this study is to review the currently available evidence that supports or rejects the above-mentioned assumption. Ultimately, it is important that SBF tests could simulate the actual physiological conditions experienced by biomaterials within the human body. Given that in vivo animal experiments provide the best pre-clinical test conditions, all studies in which both in vitro apatite forming ability and in vivo performance of two or more biomaterials are compared were found by searching the literature. From all studies that satisfied the inclusion criteria (33), in 25 studies in vitro apatite-forming ability could predict the relative performance of the tested biomaterials in vivo. In 8 studies, in vitro performance did not correctly predict the relative in vivo performance. In majority of failure cases (i.e. 5/8), none of the compared biomaterials formed apatite, while all compared biomaterials showed bioactive behavior in vivo. It is therefore concluded that, in majority of cases, the SBF immersion test has been successful in predicting the relative performance of biomaterials in vivo. However, the details of the test protocols and the (expected) mechanisms of bioactivity of tested biomaterials should be carefully considered in the design of SBF immersion tests and in interpretation of their results. Certain guidelines are devised based on the results of this review for the design of SBF immersion test protocols and interpretation of the test results. These guidelines could help in designing better SBF test protocols that have better chances of predicting the bioactivity of biomaterials for potential application in clinical orthopedics.

Transformation methods for estimation of subject-specific scapular muscle attachment sites.

The parameters that describe the soft tissue structures are among the most important anatomical parameters for subject-specific biomechanical modelling. In this paper, we study one of the soft tissue parameters, namely muscle attachment sites. Two new methods are proposed for transformation of the muscle attachment sites of any reference scapula to any destination scapula based on four palpable bony landmarks. The proposed methods as well as one previously proposed method have been applied for transformation of muscle attachment sites of one reference scapula to seven other scapulae. The transformation errors are compared among the three methods. Both proposed methods yield significantly less (p < 0.05) prediction error as compared to the currently available method. Furthermore, we investigate whether there exists a reference scapula that performs significantly better than other scapulae when used for transformation of muscle attachment sites. Seven different scapulae were used as reference scapula and their resulting transformation errors were compared with each other. In the considered statistical population, no such a thing as an ideal scapula was found. There was, however, one outlier scapula that performed significantly worse than the other scapulae when used as a reference. The effect of perturbations in both muscle attachment sites and other muscle properties is studied by comparing muscle force predictions of a musculoskeletal model between perturbed and non-perturbed versions of the model. It is found that 10 mm variations in muscle attachments have more significant effect on muscle force predictions than 10% variations in any of the other four analysed muscle properties.

How accurately can we predict the fracture load of the proximal femur using finite element models?

BACKGROUND: Current clinical methods for fracture prediction rely on two-dimensional imaging methods such as dual-energy X-ray absorptiometry and have limited predictive value. Several researchers have tried to integrate three-dimensional imaging techniques with the finite element (FE) method to improve the accuracy of fracture predictions. Before FE models could be used in clinical settings, a thorough validation of their accuracy is required. In this paper, we try to evaluate the current state of accuracy of subject-specific FE models that are used for prediction of the fracture load of proximal femora. METHODS: All the studies that have used FE for prediction of fracture load and have compared the predicted fracture load with experimentally measured fracture loads in vitro are identified through a systematic search of the literature. A quantitative analysis of the results of those studies has been carried out to determine the absolute prediction error, percentage error, and linear correlations between predicted and measured fracture loads. FINDINGS: The reported coefficients of determination (R(2)) vary between 0.773 and 0.96 while the percentage error in prediction of fracture load varies between 5 and 46% with most studies reporting percentage errors between 10 and 20%. INTERPRETATION: We conclude that FE models, which are currently used only experimentally, are in general more accurate than clinically used fracture risk assessment techniques. However, the accuracy of FE models depends on the details of their modeling methodologies. Therefore, modeling procedures need to be optimized and standardized before FE could be used in clinical settings.

Bone regeneration performance of surface-treated porous titanium.

The large surface area of highly porous titanium structures produced by additive manufacturing can be modified using biofunctionalizing surface treatments to improve the bone regeneration performance of these otherwise bioinert biomaterials. In this longitudinal study, we applied and compared three types of biofunctionalizing surface treatments, namely acid-alkali (AcAl), alkali-acid-heat treatment (AlAcH), and anodizing-heat treatment (AnH). The effects of treatments on apatite forming ability, cell attachment, cell proliferation, osteogenic gene expression, bone regeneration, biomechanical stability, and bone-biomaterial contact were evaluated using apatite forming ability test, cell culture assays, and animal experiments. It was found that AcAl and AnH work through completely different routes. While AcAl improved the apatite forming ability of as-manufactured (AsM) specimens, it did not have any positive effect on cell attachment, cell proliferation, and osteogenic gene expression. In contrast, AnH did not improve the apatite forming ability of AsM specimens but showed significantly better cell attachment, cell proliferation, and expression of osteogenic markers. The performance of AlAcH in terms of apatite forming ability and cell response was in between both extremes of AnH and AsM. AcAl resulted in significantly larger volumes of newly formed bone within the pores of the scaffold as compared to AnH. Interestingly, larger volumes of regenerated bone did not translate into improved biomechanical stability as AnH exhibited significantly better biomechanical stability as compared to AcAl suggesting that the beneficial effects of cell-nanotopography modulations somehow surpassed the benefits of improved apatite forming ability. In conclusion, the applied surface treatments have considerable effects on apatite forming ability, cell attachment, cell proliferation, and bone ingrowth of the studied biomaterials. The relationship between these properties and the bone-implant biomechanics is, however, not trivial.

Open forward and inverse problems in theoretical modeling of bone tissue adaptation.

Theoretical modeling of bone tissue adaptation started several decades ago. Many important problems have been addressed in this area of research during the last decades. However, many important questions remain unanswered. In this paper, an overview of open problems in theoretical modeling of bone tissue adaptation is presented. First, the principal elements of bone tissue adaptation models are defined and briefly reviewed. Based on these principal elements, four categories of open problems are identified. Two of these categories primarily include forward problems, while two others include inverse problems. In every one of the identified categories, important open problems are highlighted and their importance is discussed. It is shown that most of previous studies on the theoretical modeling of bone tissue adaptation have been focused on the problems of the first category and not much is done in three other categories. The paper tries to highlight these potentially important problems that have been so far largely overlooked and to inspire new avenues of research.

Full-field strain measurement and fracture analysis of rat femora in compression test.

There is a growing interest in studying the fracture behavior of bones, primarily due to the increasing societal burden of osteoporotic fractures. In addition, bone is one of the most important biological materials whose fracture behavior is not yet well understood. This is partly due to the fact that bone is a complex hierarchical material, and exhibits heterogeneous, anisotropic, and viscoelastic mechanical behavior. Understanding the fracture behavior of such a complex material requires application of a full-field strain measurement technique. Digital image correlation (DIC) is a relatively new full-field strain measurement technique that can be used for measurement of 3D surface strains during mechanical testing of different types of bones. In this study, we use the DIC technique to measure the surface strains during compression testing of two groups of rat femora. The first group of femora was harvested from young animals (12 weeks), while the second group was harvested from more mature animals (26 weeks). The surface strains are measured both in the linear range and close to the fracture. Using the measured data, we assess two strain-based fracture prediction criteria, namely equivalent strain fracture criterion and fracture limit diagram, to determine whether they can consistently predict the onset of fracture. The maximum load is measured to be 296±22 N (mean±SD) for young animals and 670±123 N for mature animals. It is shown that fracture in the vast majority of cases occurs in the area of maximum tensile strain. The equivalent strain fracture criterion predicts that the fracture occurs when the equivalent strain reaches 1.04±0.02% (average±SD) for young animals and 1.39±0.24% for mature animals. The fracture limit diagram predicts that the fracture occurs once the sum of major and minor principal surface strains reaches 0.63±0.23% for young animals and -0.63±0.30% for mature animals. Based on these numbers and consistency of the criteria with the strain values recorded at the fracture locations, it is concluded that the equivalent strain fracture criterion tends to be more consistent among the tested specimens.

Repeatability of digital image correlation for measurement of surface strains in composite long bones.

Digital image correlation (DIC) can measure full-field surface strains during mechanical testing of hard and soft tissues. When compared to traditional methods, such as strain gauges, DIC offers larger validation data (∼50,000 points) for, e.g., finite element models. Our main aim was to evaluate the repeatability of surface strain measurements with DIC during compressive testing of composite femurs mimicking human bones. We also studied the similarity of the composite femur samples using CT. Composite femurs were chosen as test material to minimize the uncertainties associated with the use of cadaveric tissues and to understand the variability of the DIC measurement itself. Six medium-sized fourth generation composite human proximal femora (Sawbones) were CT imaged and mechanically tested in stance configuration. The force-displacement curves were recorded and the 3D surface strains were measured with DIC on the anterior surface of the femurs. Five femurs fractured at the neck-trochanter junction and one at the site below the minor trochanter. CT image of this bone showed an air cavity at the initial fracture site. All femurs fractured through a sudden brittle crack. The fracture force for the composite bones was 5751±650N (mean±SD). The maximum von Mises strain during the fractures was 2.4±0.8%. Noise in one experiment was 5-30µÎµ. When applied loads were equalized the variation in strains between the bones was 20-25%, and when the maximum strains were equalized, variation in the other regions was 5-10%. DIC showed that the ability of nominally identical composite bones to bear high strains and loads before fracturing may vary between the samples.

Effects of muscle fatigue on the ground reaction force and soft-tissue vibrations during running: a model study.

A modeling approach is used in this paper to study the effects of fatigue on the ground reaction force (GRF) and the vibrations of the lower extremity soft tissues. A recently developed multiple degrees-of-freedom mass-spring-damper model of the human body during running is used for this purpose. The model is capable of taking the muscle activity into account by using a nonlinear controller that tunes the mechanical properties of the soft-tissue package based on two physiological hypotheses, namely, "constant force" and "constant vibration." In this study, muscle fatigue is implemented in the model as the gradual reduction of the ability of the controller to tune the mechanical properties of the lower body soft-tissue package. Simulations are carried out for various types of footwear in both pre- and postfatigue conditions. The simulation results show that the vibration amplitude of the lower body soft-tissue package may considerably increase (up to 20%) with muscle fatigue, while the effects of fatigue on the GRF are negligible. The results of this modeling study are in line with the experimental studies that found muscle fatigue does not significantly change the GRF peaks, but may increase the level of soft-tissue vibrations (particularly for hard shoes). A major contribution of the current study is the formulation of a hypothesis about how the central nervous system tunes the muscle properties after fatigue.

Computational load estimation of the femur.

The density distribution and, thus, mechanical properties of long bones such as the femur are dependent on their loading. Many bone tissue adaptation theories are proposed to describe the density distribution that results from a given set of loading parameters. It is relatively easy to measure the density distribution of long bones, for example, using Computed Tomography (CT). However, there is no easy non-invasive method for in-vivo measurement of musculoskeletal loads. It is therefore interesting to investigate whether or not it is possible to predict the musculoskeletal loads that have resulted in a certain measured density distribution using bone tissue adaptation models. An inverse problem has to be solved for that purpose. In this paper, we use Artificial Neural Networks (ANNs) to solve the associated inverse problem and estimate the loading parameters that have resulted in the CT-measured three-dimensional density distribution of a proximal femur. An ANN is trained using a dataset generated by solving the forward tissue adaptation model for a large number of loading parameters. Before training the ANN with the generated training dataset, a Gaussian noise component is added to the density distribution. This improves the robustness of the trained ANN against deviations of the measured density distribution from the predictions of the forward bone tissue adaptation model. It is shown that the proposed technique is capable of predicting loading parameters that result in a density distribution close to the measured density distribution.