Case Studies of a Simulation Workflow to Improve Bone Healing Assessment in Impending Non-Unions.

Background: The healing potential of a fracture is determined by mechanical and biological factors. Simulation-based workflows can help assess these factors to assist in predicting non-unions. The aim of this study was the introduction of two use cases for a novel patient-specific simulation workflow based on clinically available information. Methods: The used software is an extension of the "Ulm Bone Healing model" and was applied in two cases with non-union development after fracture fixation to show its principal feasibility. The clinical and radiographic information, starting from initial treatment, were used to feed the simulation process. Results: The simulation predicted non-union development and axial deviation in a mechanically driven non-union. In the case of a biological non-union, a slow, incomplete healing course was correctly identified. However, the time offset in callus bridging was discordant between the simulation and the distinctly slower healing response in the clinical case. Conclusions: The simulation workflow presented in the two clinical use cases allowed for the identification of fractures at risk for impending non-union immediately after the initial fixation based on available clinical and radiographic information. Further validation in a large non-union cohort is needed to increase the model's precision, especially in biologically challenging cases, and show its validity as a screening instrument.

[Non-unions of the upper extremities]. / Pseudarthrosen der oberen Extremität.

The diagnosis and treatment of non-unions still represents an interdisciplinary challenge. Therefore, prevention, early detection and specific treatment are of great importance. Non-unions of the upper extremities, although less common than that of the lower extremities, requires special attention for successful treatment due to the central role of the shoulder girdle and arm in day to day activities. Successful treatment of non-unions requires a comprehensive evaluation of the patient's medical history, a thorough clinical examination and in particular radiological imaging. In order to effectively treat the pseudarthrosis it is crucial to distinguish between pseudarthroses that are suspected to be due to infections and those that are not. This article presents a treatment algorithm for managing both pseudarthrosis due to infection and pseudarthrosis without infection in the upper extremities.

AZU1: a new promising marker for infection in orthopedic and trauma patients?

Early and reliable detection of infection is vital for successful treatment. Serum markers such as C-reactive protein (CRP) and procalcitonin (PCT) are known to increase with a time lag. Azurocidin 1 (AZU1) has emerged as a promising marker for septic patients, but its diagnostic value in orthopedic and trauma patients remains unexplored. Between July 2020 and August 2023, all patients necessitating inpatient treatment for periprosthetic joint infection (PJI), peri-implant infection (II), soft tissue infection, chronic osteomyelitis, septic arthrodesis, bone non-union with and without infection were enrolled. Patients undergoing elective total joint arthroplasty (TJA) served as the control group. Blood samples were collected and analyzed for CRP, white blood cell count (WBC), PCT, and AZU1. Based on the inclusion and exclusion criteria 222 patients were included in the study (trauma = 38, soft tissue infection = 75, TJA = 33, PJI/II = 39, others = 37). While sensitivity and specificity were comparably high for AZU1 (0.734/0.833), CRP and PCT had higher specificity (0.542/1 and 0.431/1, respectively), and WBC a slightly higher sensitivity (0.814/0.455) for septic conditions. Taken together, the area under the curve (AUC) showed the highest accuracy for AZU1 (0.790), followed by CRP (0.776), WBC (0.641), and PCT (0.656). The Youden-Index was 0.57 for AZU1, 0.54 for CRP, 0.27 for WBC, and 0.43 for PCT. Elevated AZU1 levels effectively distinguished patients with a healthy condition from those suffering from infection. However, there is evidence suggesting that trauma may influence the release of AZU1. Additional research is needed to validate the diagnostic value of this new biomarker and further explore its potential clinical applications.

[Nonunions after intertrochanteric and subtrochanteric femoral fractures]. / Pseudarthrosen nach per- und subtrochantären Femurfrakturen.

INTRODUCTION: The overall frequency of proximal femoral fractures means that we are repeatedly confronted with failed healing and implant failure, despite a relatively low nonunion rate especially in intertrochanteric fractures (< 5%). The aim of this paper is to present our approach to treating these nonunions of the proximal femur and discuss the treatment results. MATERIAL AND METHODS: Between 2009 and 2023, patients with nonunion of the proximal femur were retrospectively identified and analyzed. Age, gender, time to revision, the Weber-Cech classification of pseudarthrosis and radiographic imaging before and after revision were analyzed. RESULTS: A total of 66 patients were analyzed. The mean age was 58 years (range 25-88 years). The overall healing rate was 88% with a mean consolidation time of 8 months (range 2-29 months). The main osteosynthesis procedures were plate osteosynthesis (n = 45, of which 44 were blade plates), and nail replacement (n = 12). Other procedures included augmentative plate osteosyntheses (n = 4), isolated cancellous bone graft (n = 2), nail dynamization (n = 2), and the use of a dynamic hip screw (n = 1). DISCUSSION: The analysis of our treatment data as well as the current literature, revealed a trend towards intramedullary revision procedures. Implants that can be used to correct the CCD angle, such as the blade plate, remain a predictable option to achieve correction, especially in nonunions with an increased degree of varus. Particularly in the subtrochanteric region, fractures can also be treated in a targeted manner by a combination of mechanical and biological methods with a reamed nail change to a larger caliber implant.

The Impact of Injury of the Tibial Nutrient Artery Canal on Type of Nonunion of Tibial Shaft Fractures: A Retrospective Computed Tomography Study.

RATIONALE AND OBJECTIVES: Blood supply is vital for sound callus formation. The tibial nutrient artery (TNA) is the main diaphyseal artery nurturing the tibial shaft. The objective is to investigate the impact of TNA canal (TNAC) injury on the development of atrophic, oligotrophic, and hypertrophic nonunion in patients with tibial shaft fractures. MATERIALS AND METHODS: Between January 2010 and December 2020, patients with a nonunion of a tibial shaft fracture were retrospectively included. Two readers independently evaluated the integrity of the TNAC and classified nonunion type. A multinomial regression model was utilized to evaluate if a TNAC injury has an impact on the type of nonunion. RESULTS: From an initial set of 385 patients with the diagnosis of a nonunion of the lower leg, a total of 60 patients could be finally included in the study. Most patients were males (78%), diabetic (95%), smokers (73%), and had an American Society of Anesthesiologists (ASA) score of 2 (72%). TNAC injury was noted in 24 patients (40%): an iatrogenic TNAC injury was observed in 13 (22%) patients, a traumatic TNAC injury in 11 (18%) patients. Most patients had a hypertrophic nonunion (29 patients (48%)), followed by an oligotrophic nonunion (24 patients (40%)) and lastly an atrophic nonunion (seven patients (11%)). The multinomial regression model showed that there was no impact of TNAC injury on the development of a specific type of non-union (p = 0.798 for oligotrophic vs. atrophic nonunion; p = 0.943 for hypertrophic vs. atrophic nonunion). Furthermore, patients were about four times more likely to develop an oligotrophic/hypertrophic nonunion rather than atrophic one (odds ratio 3.75 and 4.25, respectively), regardless of the presence of a TNAC injury. CONCLUSION: In the evaluated patient cohort with tibial shaft fractures, we could not find a statistically significant association between TNAC injury and type of nonunion. However, patients were almost four times more likely to develop oligotrophic or hypertrophic nonunion rather than an atrophic one although common risk factors for impaired (micro)vascular blood supply were highly prevalent in the study group. Multicenter studies with a larger number of atrophic nonunions are warranted to further evaluate this result.

Increased Age, Cardiovascular Comorbidities, COPD, and Diabetes mellitus Determine Excess Mortality in Trauma Patients with SSI or Chronic Wounds. / Erhöhtes Alter, kardiovaskuläre Nebenerkrankungen, COPD und Diabetes mellitus bedingen eine Übersterblichkeit in der septischen Unfallchirurgie.

BACKGROUND: The constant aging of the German population leads, among other things, to an increase in the average age of hospitalised patients. In association with the reduced physiological reserve and the endogenous defence mechanism, this results in an increased susceptibility to infections and complications. Compared with that in trauma patients, the age distribution in patients with surgical site infections (SSI) has been poorly studied. Studies on the impact of age, diverse underlying diseases and influence of alcohol and nicotine consumption on the mortality risk in patients with SSI are limited. METHODS: In 2014/15 (Exam 1), 345 patients with SSI were included in this study. The 3-year follow-up examination was performed in 2017/18 (Exam 2). The questionnaires (Exams 1 and 2) assessed demographic parameters, comorbidities, medication use, alcohol and nicotine consumption, and different risk factors related to morbidity. The mortality risk in patients with SSI was calculated as a function of various risk factors (age, comorbidities, medication intake, and noxious agents). Furthermore, the development of the patients' age in the trauma department, especially of those with SSI, were evaluated between 2010 and 2019. RESULTS: In 2014/15 (Exam 1), 345 patients were included in the study. Of these, 274 (79.4%) were contacted by telephone in 2017/18. Thirty-six (10.4%) declined to be questioned again. Twenty (8.4%) of the 238 remaining participants had already died, resulting in 218 patients (63.2%) re-participating. From 2010 (n = 492) to 2019 (n = 885), the number of patients with SSI increased by 79.9%, especially those aged < 65 years. After the age of 60 years, the mortality risk increased rapidly (60 years: 0.0377 vs 70 years: 0.1395); the mortality risk of a 60-year-old patient with SSI was equal to that of an 80-year-old in the general population. Nicotine (p = 0.93) and alcohol consumption (p = 0.344) had no significant effect on mortality, whereas history of cardiac disease (p = 0.01), chronic obstructive pulmonary disease (COPD) (p = 0.01), diabetes mellitus (p = 0.05) and peripheral artery disease (p = 0.01) were associated with a significant increase in the mortality risk. CONCLUSION: Age, pre-existing cardiac conditions, as well as COPD, diabetes mellitus and peripheral artery disease are associated with a significantly increased mortality risk in patients with SSI. Thus, an exponential increase in mortality risk was found between the ages of 50 and 60 years, with the mortality risk of a 60-year-old patient with SSI being equivalent to that of an 80-year-old in the general population.

Automated Artificial Intelligence-Based Assessment of Lower Limb Alignment Validated on Weight-Bearing Pre- and Postoperative Full-Leg Radiographs.

The assessment of the knee alignment using standing weight-bearing full-leg radiographs (FLR) is a standardized method. Determining the load-bearing axis of the leg requires time-consuming manual measurements. The aim of this study is to develop and validate a novel algorithm based on artificial intelligence (AI) for the automated assessment of lower limb alignment. In the first stage, a customized mask-RCNN model was trained to automatically detect and segment anatomical structures and implants in FLR. In the second stage, four region-specific neural network models (adaptations of UNet) were trained to automatically place anatomical landmarks. In the final stage, this information was used to automatically determine five key lower limb alignment angles. For the validation dataset, weight-bearing, antero-posterior FLR were captured preoperatively and 3 months postoperatively. Preoperative images were measured by the operating orthopedic surgeon and an independent physician. Postoperative images were measured by the second rater only. The final validation dataset consisted of 95 preoperative and 105 postoperative FLR. The detection rate for the different angles ranged between 92.4% and 98.9%. Human vs. human inter-(ICCs: 0.85−0.99) and intra-rater (ICCs: 0.95−1.0) reliability analysis achieved significant agreement. The ICC-values of human vs. AI inter-rater reliability analysis ranged between 0.8 and 1.0 preoperatively and between 0.83 and 0.99 postoperatively (all p < 0.001). An independent and external validation of the proposed algorithm on pre- and postoperative FLR, with excellent reliability for human measurements, could be demonstrated. Hence, the algorithm might allow for the objective and time saving analysis of large datasets and support physicians in daily routine.

Contribution to the 3R Principle: Description of a Specimen-Specific Finite Element Model Simulating 3-Point-Bending Tests in Mouse Tibiae.

Bone mechanical properties are classically determined by biomechanical tests, which normally destroy the bones and disable further histological or molecular analyses. Thus, obtaining biomechanical data from bone usually requires an additional group of animals within the experimental setup. Finite element models (FEMs) may non-invasively and non-destructively simulate mechanical characteristics based on material properties. The present study aimed to establish and validate an FEM to predict the mechanical properties of mice tibiae. The FEM was established based on µCT (micro-Computed Tomography) data of 16 mouse tibiae. For validating the FEM, simulated parameters were compared to biomechanical data obtained from 3-point bending tests of the identical bones. The simulated and the measured parameters correlated well for bending stiffness (R2 = 0.9104, p < 0.0001) and yield displacement (R2 = 0.9003, p < 0.0001). The FEM has the advantage that it preserves the bones' integrity, which can then be used for other analytical methods. By eliminating the need for an additional group of animals for biomechanical tests, the established FEM can contribute to reducing the number of research animals in studies focusing on bone biomechanics. This is especially true when in vivo µCT data can be utilized where multiple bone scans can be performed with the same animal at different time points. Thus, by partially replacing biomechanical experiments, FEM simulations may reduce the overall number of animals required for an experimental setup investigating bone biomechanics, which supports the 3R (replace, reduce, and refine) principle.

[Artificial intelligence and novel approaches for treatment of non-union in bone : From established standard methods in medicine up to novel fields of research]. / Künstliche Intelligenz und Ausblick auf Anwendungsfelder in der Pseudarthrosentherapie : Von etablierten Standardmethoden in der Medizin hin zu neuen Forschungsfeldern.

Methods of artificial intelligence (AI) have found applications in many fields of medicine within the last few years. Some disciplines already use these methods regularly within their clinical routine. However, the fields of application are wide and there are still many opportunities to apply these new AI concepts. This review article gives an insight into the history of AI and defines the special terms and fields, such as machine learning (ML), neural networks and deep learning. The classical steps in developing AI models are demonstrated here, as well as the iteration of data rectification and preparation, the training of a model and subsequent validation before transfer into a clinical setting are explained. Currently, musculoskeletal disciplines implement methods of ML and also neural networks, e.g. for identification of fractures or for classifications. Also, predictive models based on risk factor analysis for prevention of complications are being initiated. As non-union in bone is a rare but very complex disease with dramatic socioeconomic impact for the healthcare system, many open questions arise which could be better understood by using methods of AI in the future. New fields of research applying AI models range from predictive models and cost analysis to personalized treatment strategies.

[Movement analysis and musculoskeletal simulation in non-union treatment-Experiences and first clinical results]. / Bewegungsanalyse und muskuloskeletale Simulation in der Pseudarthrosentherapie ­ Erfahrungen und erste klinische Ergebnisse.

BACKGROUND: The mechanical boundary conditions of the non-union and osteosynthetic construct are a key determinant of fracture healing after revision surgery. Aim of this study was to introduce a movement analysis and simulation workflow to determine the mechanical conditions during non-union healing in vivo. MATERIAL AND METHODS: On an individual case basis after non-union revision surgery we performed an accelerometry-based movement analysis. The results were then used as input for a musculoskeletal simulation of the non-union, osteosynthetic construct as well as adjacent joints mechanical boundary conditions. RESULTS: A total of 13 patients were analyzed with our new workflow. The introduced protocol allows an in vivo determination of the mechanical boundary conditions. On clinical follow-up all patients showed radiographic consolidation of the non-union. CONCLUSION: The introduced workflow allows a clinically applicable determination of the mechanical boundary conditions of fracture and non-union healing. Further studies can now determine the effect of the introduced technique for mechanically optimized postoperative aftercare regimes as well as biomechanically adapted surgical treatment.

[Atrophic sternum nonunion-Case report based on the nonunion scoring system]. / Atrophe Pseudarthrose des Sternums ­ ein Fallbericht orientiert am Non Union Scoring System.

Nonunions after median sternotomy are rare and usually respond well to surgical treatment. Recalcitrant nonunions despite surgical treatment require a comprehensive mechanical and biological treatment strategy to achieve an adequate functional result for the patient. We demonstrate the case of a 4-year recalcitrant atrophic nonunion. Through a surgical approach guided by the criteria of the nonunion scoring system (NUSS) successful healing was achieved. Our strategy as well as the treatment course are presented.

Risk of malnutrition in orthopedic trauma patients with surgical site infections is associated with increased morbidity and mortality - a 3-year follow-up study.

BACKGROUND: Malnutrition is a worldwide problem which can result in prolonged hospitalization from complications such as poor wound healing and increased morbidity. There is increasing evidence of the effect of risk of malnutrition (ROM) on outcomes in orthopedic surgical patients. However, there is little data on the effect of nutritional status on clinical outcomes in orthopedic trauma patients with surgical site infections (SSI). Therefore, our aim was to investigate how malnutrition risk affects clinical outcomes in a prospective cohort of orthopedic trauma patients with SSI. METHODS: The study included 345 patients who underwent surgery due to SSI at a level 1 trauma center. All patients were evaluated on their nutritional status as assessed by the Nutritional Risk Screening in 2014/15 and 2017/18. 238 (69.0%) datasets were available for the follow-up analysis. Twenty patients (8.4%) had died, resulting in 218 patients. Outcomes investigated included comorbidities, medication intake, destination of discharge, degree of mobility, support for procuring food, mortality risk and quality of life. RESULTS: 32.8% were at risk of malnutrition (ROM) at EXAM1. Female patients had a higher ROM than males (p < 0.05). Patients with ROM had more comorbidities (p < 0.001), an increased need for medication intake (p < 0.001), a decreased level of mobility (p < 0.001) and increased need of support in procuring food (p < 0.001). The destination of discharge was independent of the nutritional status (p = 0.641). Twenty (8.4%) of the available 238 patients had died during follow-up time period, resulting in a 6.2-times higher risk of mortality in patients with ROM. EQ-5D revealed that mobility, self-supply and usual activities of daily living were increased in well-nourished patients (p < 0.001). CONCLUSION: ROM in orthopedic trauma patients with SSI is associated with an increased number of comorbidities and need for medication intake, a decrease in mobility and a higher dependency for food acquisition. Patients at ROM exhibited a 6.2-times higher mortality rate than well-nourished patients. EQ-5D evaluation showed better mobility, self-supply, and activity of daily living in well-nourished patients. We therefore strongly recommend supplementing patients with ROM with a specific diet during and after discharge from the hospital in order to reduce postoperative complications and long-term mortality.

Injury of the Tibial Nutrient Artery Canal during External Fixation for Lower Extremity Fractures: A Computed Tomography Study.

The tibial nutrient artery (TNA) is the major diaphyseal artery of the tibia supplying two thirds of the inner osseous cortex. Hence, iatrogenic injury of the TNA endangers the integrity of the tibial blood supply and may compromise fracture healing. The incidence of its injury in the setting of external fixation for lower limb fractures has not been previously investigated. The aim of this study was to evaluate the incidence of TNA injury in the context of external fixation and to characterize the topography of the fixator pins in relation to the TNA canal (TNAC). Patients who underwent external fixation for distal femoral fractures and for tibial (proximal, shaft, and distal) fractures and had a postoperative computed tomography study were retrospectively included. The following parameters were retrieved: 1) Pin characteristics (orientation and cortical position of the pins), 2) The anatomic relationship between the TNAC and external fixation pin (topography above/below and at the level of the TNAC, and the distance between the pin and medial tibial plateau and/or the medial malleolus), and 3) The incidence of TNAC injury (complete/partial disruption of TNA lumen). A total of 105 patients with 214 tibial pins were analyzed. In 27 patients (26%), the TNAC was completely injured by the pins of the external fixator. In 13 patients (12%), the TNAC was partially injured. Of the 214 analyzed pins, 85 pins (40%) were located at the level of the TNAC (the TNAC and the pin are seen on the same axial slice). Most pins that were applied at the level of the TNAC belonged to a knee-bridging external fixator. Of those, ninety-three percent of the pins were anteromedially applied according to published surgical guidelines. Six percent of the pins were applied through the tibial crest and 1% anterolaterally. Of those 85 pins, 42 pins (49%) injured the TNAC at least partially. Based on the analyzed pins and the incidence of partial and complete injury of the TNAC, we observed that the tibial segment at which the tibial nutrient artery is endangered was located approximately (95% CI: 13-15 cm) from the medial tibia plateau and (95% CI: 22-25 cm) from the medial malleolus. Thus, TNAC injury by external fixation pins in the context of lower limb fractures can be considered common. Almost half of the pins applied at the middle third of the tibia injured the TNA, despite adherence to published surgical guidelines for external fixation. When possible, pin application at the middle third of tibia should be avoided to circumvent iatrogenic injury of the TNA and to safeguard tibial blood supply.

E-vapor aerosols do not compromise bone integrity relative to cigarette smoke after 6-month inhalation in an ApoE-/- mouse model.

Cigarette smoke (CS) exposure is one of the leading risk factors for human health. Nicotine-containing inhalable products, such as e-cigarettes, can effectively support tobacco harm reduction approaches. However, there are limited comparative data on the effects of the aerosols generated from electronic vapor products (e-vapor) and CS on bone. Here, we report the effects of e-vapor aerosols and CS on bone morphology, structure, and strength in a 6-month inhalation study. Eight-week-old ApoE-/- mice were exposed to aerosols from three different e-vapor formulations-CARRIER (propylene glycol and vegetable glycerol), BASE (CARRIER and nicotine), TEST (BASE and flavor)-to CS from 3R4F reference cigarettes at matched nicotine concentrations (35 µg/L) or to fresh air (Sham) (N = 10 per group). Tibiae were analyzed for bone morphology by µCT imaging, biomechanics by three-point bending, and by histological analysis. CS inhalation caused a significant decrease in cortical and total bone volume fraction and bone density relative to e-vapor aerosols. Additionally, CS exposure caused a decrease in ultimate load and stiffness. In contrast, bone structural and biomechanical parameters were not significantly affected by e-vapor aerosol or Sham exposure. At the dissection time point, there was no significant difference in body weight or tibia bone weight or length among the groups. Histological findings revealed microcracks in cortical bone areas among all exposed groups compared to Sham control. In conclusion, because of the bone-preserving effect of e-vapor aerosols relative to CS exposure, e-vapor products could potentially constitute less harmful alternatives to cigarettes in situations in which bone health is of importance.

The Anatomy of the Tibial Nutrient Artery Canal-An Investigation of 106 Patients Using Multi-Detector Computed Tomography.

Radiologic evaluation of the tibial nutrient artery is clinically important as disruption of tibial blood supply is a risk factor for delayed or non-union of tibial fractures. Damage to the tibial nutrient artery canal (TNAC) may occur by a traversing fracture or iatrogenic cause in the context of pin/screw placement. Furthermore, TNAC could be misdiagnosed as a stress fracture. The aim of this study was to characterize the normal anatomy of TNAC and to delineate its gender and side-specific differences. Patients who underwent contrast-enhanced computed-tomography encompassing the pelvis and lower extremities were included. TNAC was identified with an external and internal foramen and a traversing intercortical canal. Various anatomical morphometrics were evaluated: total number of nutrient canals, angular position of the outer and inner nutrient foramina, absolute and relative position of the nutrient foramina, as well as the intercortical canal length with respect to tibial length. The majority of patients of both genders had only one tibial nutrient canal, multiple canals or complete absence were rare. In most cases, the outer nutrient foramen was found on the posterolateral aspect of the upper-third of tibia at about 32% of tibial length; the inner foramen was found at the middle third of the tibia (41% of tibial length). The course of nutrient canal was mostly cranio-caudal with a small, but significant difference in relative canal length: 8.5% vs. 10% of tibial length for females and males, respectively. The angular location of the outer and inner foramen was between 20-30° and did not reveal a statistically significant difference between genders. No statistically significant side specific differences were found for all analyzed parameters and both genders. The clinical relevance of this anatomical study pertains to establishing "safe corridors" of pin/screw insertion in the context of surgical management of tibial fractures in order to avoid iatrogenic disruption of tibial blood supply.

Donor Site Location Is Critical for Proliferation, Stem Cell Capacity, and Osteogenic Differentiation of Adipose Mesenchymal Stem/Stromal Cells: Implications for Bone Tissue Engineering.

Human adipose mesenchymal stem/stromal cells (Ad-MSCs) have been proposed as a suitable option for bone tissue engineering. However, donor age, weight, and gender might affect the outcome. There is still a lack of knowledge of the effects the donor tissue site might have on Ad-MSCs function. Thus, this study investigated proliferation, stem cell, and osteogenic differentiation capacity of human Ad-MSCs obtained from subcutaneous fat tissue acquired from different locations (abdomen, hip, thigh, knee, and limb). Ad-MSCs from limb and knee showed strong proliferation despite the presence of osteogenic stimuli, resulting in limited osteogenic characteristics. The less proliferative Ad-MSCs from hip and thigh showed the highest alkaline phosphatase (AP) activity and matrix mineralization. Ad-MSCs from the abdomen showed good proliferation and osteogenic characteristics. Interestingly, the observed differences were not dependent on donor age, weight, or gender, but correlated with the expression of Sox2, Lin28A, Oct4α, and Nanog. Especially, low basal Sox2 levels seemed to be pivotal for osteogenic differentiation. Our data clearly show that the donor tissue site affects the proliferation and osteogenic differentiation of Ad-MSCs significantly. Thus, for bone tissue engineering, the donor site of the adipose tissue from which the Ad-MSCs are derived should be adapted depending on the requirements, e.g., cell number and differentiation state.

Sclerostin Neutralizing Antibody Treatment Enhances Bone Formation but Does Not Rescue Mechanically Induced Delayed Healing.

During bone healing, tissue formation processes are governed by mechanical strain. Sost/sclerostin, a key Wnt signaling inhibitor and mechano-sensitive pathway, is downregulated in response to mechanical loading. Sclerostin neutralizing antibody (SclAb) increases bone formation. Nevertheless, it remains unclear whether sclerostin inhibition can rescue bone healing in situations of mechanical instability, which otherwise delay healing. We investigated SclAb's influence on tissue formation in a mouse femoral osteotomy, stabilized with rigid or semirigid external fixation. The different fixations allowed different magnitudes of interfragmentary movement during weight bearing, thereby influencing healing outcome. SclAb or vehicle (veh) was administeredand bone healing was assessed at multiple time points up to day 21 postoperatively by in vivo micro-computed tomography, histomorphometry, biomechanical testing, immunohistochemistry, and gene expression. Our results show that SclAb treatment caused a greater bone volume than veh. However, SclAb could not overcome the characteristic delayed healing of semirigid fixation. Indeed, semirigid fixation resulted in delayed healing with a prolonged endochondral ossification phase characterized by increased cartilage, lower bone volume fraction, and less bony bridging across the osteotomy gap than rigid fixation. In a control setting, SclAb negatively affected later stages of healing under rigid fixation, evidenced by the high degree of endosteal bridging at 21 days in the rigid-SclAb group compared with rigid-veh, indicating delayed fracture callus remodeling and bone marrow reconstitution. Under rigid fixation, Sost and sclerostin expression at the gene and protein level, respectively, were increased in SclAb compared with veh-treated bones, suggesting a negative feedback mechanism. Our results suggest that SclAb could be used to enhance overall bone mass but should be carefully considered in bone healing. SclAb may help to increase bone formation early in the healing process but not during advanced stages of fracture callus remodeling and not to overcome delayed healing in semirigid fixation. © 2018 American Society for Bone and Mineral Research.

Osteoblast migration in vertebrate bone.

Bone formation, for example during bone remodelling or fracture repair, requires mature osteoblasts to deposit bone with remarkable spatial precision. As osteoblast precursors derive either from circulation or resident stem cell pools, they and their progeny are required to migrate within the three-dimensional bone space and to navigate to their destination, i.e. to the site of bone formation. An understanding of this process is emerging based on in vitro and in vivo studies of several vertebrate species. Receptors on the osteoblast surface mediate cell adhesion and polarization, which induces osteoblast migration. Osteoblast migration is then facilitated along gradients of chemoattractants. The latter are secreted or released proteolytically by several cell types interacting with osteoblasts, including osteoclasts and vascular endothelial cells. The positions of these cellular sources of chemoattractants in relation to the position of the osteoblasts provide the migrating osteoblasts with tracks to their destination, and osteoblasts possess the means to follow a track marked by multiple chemoattractant gradients. In addition to chemotactic cues, osteoblasts sense other classes of signals and utilize them as landmarks for navigation. The composition of the osseous surface guides adhesion and hence migration efficiency and can also provide steering through haptotaxis. Further, it is likely that signals received from surface interactions modulate chemotaxis. Besides the nature of the surface, mechanical signals such as fluid flow may also serve as navigation signals for osteoblasts. Alterations in osteoblast migration and navigation might play a role in metabolic bone diseases such as osteoporosis.

Extremely low frequency pulsed electromagnetic fields cause antioxidative defense mechanisms in human osteoblasts via induction of •O2- and H2O2.

Recently, we identified a specific extremely low-frequency pulsed electromagnetic field (ELF-PEMF) that supports human osteoblast (hOBs) function in an ERK1/2-dependent manner, suggesting reactive oxygen species (ROS) being key regulators in this process. Thus, this study aimed at investigating how ELF-PEMF exposure can modulate hOBs function via ROS. Our results show that single exposure to ELF-PEMF induced ROS production in hOBs, without reducing intracellular glutathione. Repetitive exposure (>3) to ELF-PEMF however reduced ROS-levels, suggesting alterations in the cells antioxidative stress response. The main ROS induced by ELF-PEMF were •O2- and H2O2, therefore expression/activity of antioxidative enzymes related to these ROS were further investigated. ELF-PEMF exposure induced expression of GPX3, SOD2, CAT and GSR on mRNA, protein and enzyme activity level. Scavenging •O2- and H2O2 diminished the ELF-PEMF effect on hOBs function (AP activity and mineralization). Challenging the hOBs with low amounts of H2O2 on the other hand improved hOBs function. In summary, our data show that ELF-PEMF treatment favors differentiation of hOBs by producing non-toxic amounts of ROS, which induces antioxidative defense mechanisms in these cells. Thus, ELF-PEMF treatment might represent an interesting adjunct to conventional therapy supporting bone formation under oxidative stress conditions, e.g. during fracture healing.