Sildenafil delays bone remodeling of fractured femora in aged mice by reducing the number and activity of osteoclasts within the callus tissue.

The elderly exhibit a reduced healing capacity after fracture, which is often associated with delayed or failed bone healing. This is due to a plethora of factors, such as an impaired bone vascular system and delayed angiogenesis. The phosphodiesterase-5 (PDE-5) inhibitor sildenafil exerts pro-angiogenic and pro-osteogenic effects. Hence, we herein investigated in aged mice whether sildenafil can improve fracture healing. For this purpose, 40 aged CD-1 mice (16-18 months) were daily treated with 5 mg/kg body weight sildenafil (n = 20) or vehicle (control, n = 20) by oral gavage. The callus tissue of their femora was analyzed at 2 and 5 weeks after fracture by X-ray, biomechanics, micro-computed tomography (µCT), histology, immunohistochemistry as well as Western blotting. These analyses revealed a significantly increased bone volume and higher ratio of callus to femoral bone diameter in sildenafil-treated mice at 5 weeks after fracture when compared to controls. This was associated with a reduced number and activity of osteoclasts at 2 weeks after fracture, most likely caused by an increased expression of osteoprotegerin (OPG). Taken together, these findings indicate that sildenafil does not improve fracture healing in the elderly but delays the process of bone remodeling most likely by reducing the number and activity of osteoclasts within the callus tissue.

[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.

Will I soon be out of my job? Quality and guideline conformity of ChatGPT therapy suggestions to patient inquiries with gynecologic symptoms in a palliative setting.

PURPOSE: The market and application possibilities for artificial intelligence are currently growing at high speed and are increasingly finding their way into gynecology. While the medical side is highly represented in the current literature, the patient's perspective is still lagging behind. Therefore, the aim of this study was to evaluate the recommendations of ChatGPT regarding patient inquiries about the possible therapy of gynecological leading symptoms in a palliative situation by experts. METHODS: Case vignettes were constructed for 10 common concomitant symptoms in gynecologic oncology tumors in a palliative setting, and patient queries regarding therapy of these symptoms were generated as prompts for ChatGPT. Five experts in palliative care and gynecologic oncology evaluated the responses with respect to guideline adherence and applicability and identified advantages and disadvantages. RESULTS: The overall rating of ChatGPT responses averaged 4.1 (5 = strongly agree; 1 = strongly disagree). The experts saw an average guideline conformity of the therapy recommendations with a value of 4.0. ChatGPT sometimes omits relevant therapies and does not provide an individual assessment of the suggested therapies, but does indicate that a physician consultation is additionally necessary. CONCLUSIONS: Language models, such as ChatGPT, can provide valid and largely guideline-compliant therapy recommendations in their freely available and thus in principle accessible version for our patients. For a complete therapy recommendation, an evaluation of the therapies, their individual adjustment as well as a filtering of possible wrong recommendations, a medical expert's opinion remains indispensable.

Wearable activity data can predict functional recovery after musculoskeletal injury: Feasibility of a machine learning approach.

Delayed functional recovery after injury is associated with significant personal and socioeconomic burden. Identification of patients at risk for a prolonged recovery after a musculoskeletal injury is thus of high relevance. The aim of the current study was to show the feasibility of using a machine learning assisted model to predict functional recovery based on the pre- and immediate post injury patient activity as measured with wearable systems in trauma patients. Patients with a pre-existing wearable (smartphone and/or body-worn sensor), data availability of at least 7 days prior to their injury, and any musculoskeletal injury of the upper or lower extremity were included in this study. Patient age, sex, injured extremity, time off work and step count as activity data were recorded continuously both pre- and post-injury. Descriptive statistics were performed and a logistic regression machine learning model was used to predict the patient's functional recovery status after 6 weeks based on their pre- and post-injury activity characteristics. Overall 38 patients (7 upper extremity, 24 lower extremity, 5 pelvis, 2 combined) were included in this proof-of-concept study. The average follow-up with available wearable data was 85.4 days. Based on the activity data, a predictive model was constructed to determine the likelihood of having a recovery of at least 50 % of the pre-injury activity state by post injury week 6. Based on the individual activity by week 3 a predictive accuracy of over 80 % was achieved on an independent test set (F1=0,82; AUC=0,86; ACC=8,83). The employed model is feasible to assess the principal risk for a slower recovery based on readily available personal wearable activity data. The model has the potential to identify patients requiring additional aftercare attention early during the treatment course, thus optimizing return to the pre-injury status through focused interventions. Additional patient data is needed to adapt the model to more specifically focus on different fracture entities and patient groups.

A pilot study on the efficacy of GPT-4 in providing orthopedic treatment recommendations from MRI reports.

Large language models (LLMs) have shown potential in various applications, including clinical practice. However, their accuracy and utility in providing treatment recommendations for orthopedic conditions remain to be investigated. Thus, this pilot study aims to evaluate the validity of treatment recommendations generated by GPT-4 for common knee and shoulder orthopedic conditions using anonymized clinical MRI reports. A retrospective analysis was conducted using 20 anonymized clinical MRI reports, with varying severity and complexity. Treatment recommendations were elicited from GPT-4 and evaluated by two board-certified specialty-trained senior orthopedic surgeons. Their evaluation focused on semiquantitative gradings of accuracy and clinical utility and potential limitations of the LLM-generated recommendations. GPT-4 provided treatment recommendations for 20 patients (mean age, 50 years ± 19 [standard deviation]; 12 men) with acute and chronic knee and shoulder conditions. The LLM produced largely accurate and clinically useful recommendations. However, limited awareness of a patient's overall situation, a tendency to incorrectly appreciate treatment urgency, and largely schematic and unspecific treatment recommendations were observed and may reduce its clinical usefulness. In conclusion, LLM-based treatment recommendations are largely adequate and not prone to 'hallucinations', yet inadequate in particular situations. Critical guidance by healthcare professionals is obligatory, and independent use by patients is discouraged, given the dependency on precise data input.

"Fall Risk Scoring" in Outpatient Gait Analysis: Validation of a New Fall Risk Assessment for Nursing Home Residents. / "Fall Risk Scoring" in der ambulanten Ganganalyse: Validierung eines neuen Sturzrisikoassessments bei Heimbewohnern.

Falls in senior home residents are common. Individual preventive training can lower the fall risk. To detect the need for training, a systematic assessment of the individual fall risk is needed. The aim of this study was thus to assess whether a fall risk score based on free field insole measurements can distinguish between an at-risk group of senior home residents and a healthy young control group. A published fall risk score was used in senior home residents over the age of 75 and a young (< 40 years) control group to determine the individual fall risk. In addition, the fall events over 12 months were assessed. Statistical analysis including ROC analysis was performed to determine the ability of the score to detect participants at heightened fall risk. In total, 18 nursing home residents and 9 young control participants were included. Of the nursing home residents, 15 had at least one fall, with a total of 37 falls recorded over 12 months. In the control group, no falls were recorded. The fall risk score was significantly different between nursing home residents and the control group (9.2 + 3.2 vs. 5.7 ± 2.2). Furthermore, the score significantly differentiated fallers from non-fallers (10.3 ± 1.8 vs. 5.2 ± 2.5), with a cut-off > 7.5 (AUC: 0.95) and a sensitivity of 86.7% (specificity 83.3%). The fall risk score is able to detect the difference between senior nursing home residents and young, healthy controls, as well as between fallers and non-fallers. Its main proof of concept is demonstrated, as based on movement data outside special gait labs, and it can simplify the risk of fall determination in geriatric nursing home residents and can now be used in further, prospective studies.

Sildenafil, a phosphodiesterase-5 inhibitor, stimulates angiogenesis and bone regeneration in an atrophic non-union model in mice.

Non-union formation represents a major complication in trauma and orthopedic surgery. The phosphodiesterase-5 (PDE-5) inhibitor sildenafil has been shown to exert pro-angiogenic and pro-osteogenic effects in vitro and in vivo. Therefore, the aim of the present study was to analyze the impact of sildenafil in an atrophic non-union model in mice. After creation of a 1.8 mm segmental defect, mice femora were stabilized by pin-clip fixation. Bone regeneration was analyzed by means of X-ray, biomechanics, photoacoustic and micro-computed tomography (µCT) imaging as well as histological, immunohistochemical and Western blot analyses at 2, 5 and 10 weeks after surgery. The animals were treated daily with either 5 mg/kg body weight sildenafil (n = 35) or saline (control; n = 35) per os. Bone formation was markedly improved in defects of sildenafil-treated mice when compared to controls. This was associated with a higher bending stiffness as well as an increased number of CD31-positive microvessels and a higher oxygen saturation within the callus tissue. Moreover, the bone defects of sildenafil-treated animals contained more tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts and CD68-positive macrophages and exhibited a higher expression of the pro-angiogenic and pro-osteogenic markers cysteine rich protein (CYR)61 and vascular endothelial growth factor (VEGF) when compared to controls. These findings demonstrate that sildenafil acts as a potent stimulator of angiogenesis and bone regeneration in atrophic non-unions.

Diclofenac, a NSAID, delays fracture healing in aged mice.

Nonsteroidal anti-inflammatory drugs (NSAIDs), such as diclofenac, belong to the most prescribed analgesic medication after traumatic injuries. However, there is accumulating evidence that NSAIDs impair fracture healing. Because bone regeneration in aged patients is subject to significant changes in cell differentiation and proliferation as well as a markedly altered pharmacological action of drugs, we herein analyzed the effects of diclofenac on bone healing in aged mice using a stable closed femoral facture model. Thirty-three mice (male n = 14, female n = 19) received a daily intraperitoneal injection of diclofenac (5 mg/kg body weight). Vehicle-treated mice (n = 29; male n = 13, female n = 16) served as controls. Fractured mice femora were analyzed by means of X-ray, biomechanics, micro computed tomography (µCT), histology and Western blotting. Biomechanical analyses revealed a significantly reduced bending stiffness in diclofenac-treated animals at 5 weeks after fracture when compared to vehicle-treated controls. Moreover, the callus tissue in diclofenac-treated aged animals exhibited a significantly reduced amount of bone tissue and higher amounts of fibrous tissue. Further histological analyses demonstrated less lamellar bone after diclofenac treatment, indicating a delay in callus remodeling. This was associated with a decreased number of osteoclasts and an increased expression of osteoprotegerin (OPG) during the early phase of fracture healing. These findings indicate that diclofenac delays fracture healing in aged mice by affecting osteogenic growth factor expression and bone formation as well as osteoclast activity and callus remodeling.

Best practice in digital orthopaedics.

Digitization in orthopaedics and traumatology is an enormously fast-evolving field with numerous players and stakeholders. It will be of utmost importance that the different groups of technologists, users, patients, and actors in the healthcare systems learn to communicate in a language with a common basis. Understanding the requirements of technologies, the potentials of digital application, their interplay, and the combined aim to improve health of patients, would lead to an extraordinary chance to improve health care. Patients' expectations and surgeons' capacities to use digital technologies must be transparent and accepted by both sides. The management of big data needs tremendous care as well as concepts for the ethics in handling data and technologies have to be established while also considering the impact of withholding or delaying benefits thereof. This review focuses on the available technologies such as Apps, wearables, robotics, artificial intelligence, virtual and augmented reality, smart implants, and telemedicine. It will be necessary to closely follow the future developments and carefully pay attention to ethical aspects and transparency.

Computer-Based Mechanobiological Fracture Healing Model Predicts Non-Union of Surgically Treated Diaphyseal Femur Fractures.

As non-unions are still common, a predictive assessment of healing complications could enable immediate intervention before negative impacts for the patient occur. The aim of this pilot study was to predict consolidation with the help of a numerical simulation model. A total of 32 simulations of patients with closed diaphyseal femoral shaft fractures treated by intramedullary nailing (PFNA long, FRN, LFN, and DePuy Synthes) were performed by creating 3D volume models based on biplanar postoperative radiographs. An established fracture healing model, which describes the changes in tissue distribution at the fracture site, was used to predict the individual healing process based on the surgical treatment performed and full weight bearing. The assumed consolidation as well as the bridging dates were retrospectively correlated with the clinical and radiological healing processes. The simulation correctly predicted 23 uncomplicated healing fractures. Three patients showed healing potential according to the simulation, but clinically turned out to be non-unions. Four out of six non-unions were correctly detected as non-unions by the simulation, and two simulations were wrongfully diagnosed as non-unions. Further adjustments of the simulation algorithm for human fracture healing and a larger cohort are necessary. However, these first results show a promising approach towards an individualized prognosis of fracture healing based on biomechanical factors.

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.

Intermittent Exposure to a 16 Hz Extremely Low Frequency Pulsed Electromagnetic Field Promotes Osteogenesis In Vitro through Activating Piezo 1-Induced Ca2+ Influx in Osteoprogenitor Cells.

Exposure to extremely low frequency pulsed electromagnetic fields (ELF-PEMF) is supposed to simulate local EMF generated during mechanical stimulation of bone and may therefore be used to improve bone regeneration. This study aimed at optimizing the exposure strategy and investigating the underlying mechanisms of a 16 Hz ELF-PEMF, previously reported to boost osteoblast function. Comparing influences of daily continuous (30 min every 24 h) and intermittent (10 min every 8 h) exposure to the 16 Hz ELF-PEMF on osteoprogenitor cells revealed that the intermittent exposure strategy enhanced the 16 Hz ELF-PEMF effects regarding cell numbers and osteogenic function. Gene expression of piezo 1 and related Ca2+ influx were significantly increased in SCP-1 cells with the daily intermittent exposure. Pharmacological inhibition of piezo 1 with Dooku 1 largely abolished the positive effect of the 16 Hz ELF-PEMF exposure on osteogenic maturation of SCP-1 cells. In summary, the intermittent exposure strategy enhanced the positive effects of 16 Hz continuous ELF-PEMF exposure in terms of cell viability and osteogenesis. This effect was shown to be mediated by an increased expression of piezo 1 and related Ca2+ influx. Thus, the intermittent exposure strategy is a promising way to further optimize the therapeutic effects of the 16 Hz ELF-PEMF regarding fracture healing or osteoporosis.

"Bring Your Own Device"-A New Approach to Wearable Outcome Assessment in Trauma.

Background and Objectives: Outcome data from wearable devices are increasingly used in both research and clinics. Traditionally, a dedicated device is chosen for a given study or clinical application to collect outcome data as soon as the patient is included in a study or undergoes a procedure. The current study introduces a new measurement strategy, whereby patients' own devices are utilized, allowing for both a pre-injury baseline measure and ability to show achievable results. Materials and Methods: Patients with a pre-existing musculoskeletal injury of the upper and lower extremity were included in this exploratory, proof-of-concept study. They were followed up for a minimum of 6 weeks after injury, and their wearable outcome data (from a smartphone and/or a body-worn sensor) were continuously acquired during this period. A descriptive analysis of the screening characteristics and the observed and achievable outcome patterns was performed. Results: A total of 432 patients was continuously screened for the study, and their screening was analyzed. The highest success rate for successful inclusion was in younger patients. Forty-eight patients were included in the analysis. The most prevalent outcome was step count. Three distinctive activity data patterns were observed: patients recovering, patients with slow or no recovery, and patients needing additional measures to determine treatment outcomes. Conclusions: Measuring outcomes in trauma patients with the Bring Your Own Device (BYOD) strategy is feasible. With this approach, patients were able to provide continuous activity data without any dedicated equipment given to them. The measurement technique is especially suited to particular patient groups. Our study's screening log and inclusion characteristics can help inform future studies wishing to employ the BYOD design.

The modified 5-item frailty index determines the length of hospital stay and accompanies with mortality rate in patients with bone and implant-associated infections after trauma and orthopedic surgery.

BACKGROUND: Bone and implant-associated infections are severe complications after trauma and orthopedic surgery. The modified 5-item frailty index (mFI-5) is an easily applicable score to predict adverse outcome after surgery. The current literature regarding mFI-5 is focused on a period of 30-days postoperative. PURPOSE: This study aims to assess the impact of frailty in orthopedic trauma patients with bone and implant-associated infections. mFI-5 was calculated from a database, which prospectively collects data about factors potentially correlated with peri- and postoperative complications since 2014. METHODS: In a level I trauma center a total of 345 patients with surgical site infections were enrolled in this study. Hereof, patients with fracture-related infections after osteosynthesis, periprosthetic joint infections of the hip and knee and post-operative osteomyelitis were included. Extensive medical baseline examination was performed in 2013/14, a three-year follow-up was organized as a telephone interview. The mFI-5 score was calculated based on the 5 factor-principle as established by Subramaniam. The nutritional status was assessed using the Nutritional Risk Screening (NRS-2002). RESULTS: 130 patients were included, whereof seven had died, resulting in 123 patients. A grouping of our patients was performed in mFI-5 = 0 (n = 46; 36,4%), mFI-5 = 1 (n = 41; 33,3%) or mFI-5 ≥ 2 (n = 36; 29,3%). Sex distribution showed 69,1% male and 30,9% female patients. Frailty did neither impact on the re-admission (p = 0,433) nor the reoperation (p = 0,327) rate in our cohort. The mortality risk nearly doubled (1,7 times) in frail patients, but did not reach significance. In hospital stay was prolonged due to frailty (12,1 ± 11,8; p = 0,004) compared to those with a mFI-5 = 0 (5,9 ± 5,1) or mFI-5 = 1 (6,9 ± 5,9). Frailty goes along with a risk of malnutrition and increases with age. CONCLUSION: The modified 5-item frailty index is not a suitable screening tool for predicting revision rate, re-admission rate, and mortality in our orthopedic trauma patient population with bone and implant-associated infections. Nevertheless, frailty is associated with an increased risk of malnutrition and increases with age.

Experiences in the Use of Motorized Intramedullary Nails after Complex Injuries to the Extremities. / Erfahrungen bei der Anwendung motorisierter Marknägel nach komplexen Extremitätenverletzungen.

The treatment of complex injuries of the extremities after comminuted fractures or non-unions is a challenging area in the field of trauma surgery. Internal, motorized implants nowadays enable a patient-oriented and progressive treatment of these cases. The present article aims to present modern treatment strategies of complex injuries of the extremities, support the use of novel, motorized intramedullary nails and provide experiences for the handling with lengthening nails or transport nails. For this purpose, the preoperative planning including selection of patients, presentation of internal lengthening and transport systems and the most important factors during preparation of the surgery are described. Moreover, critical steps during the implantation of motorized nails and also during potential follow-up interventions are highlighted and the postoperative protocol including precise recommendations for the transport und consolidation phase are provided. Finally, the experiences are illustrated by presentation of the four different cases. The use of internal, motorized implants represents the latest step in the treatment of complex injuries of the extremities. These implants improve the quality of life and the authors recommend its use. However, these implants require a high expertise and adaption of established treatment protocols in these challenging trauma cases. Follow-up analyses with a considerably large number of cases are necessary and the research on implants to solve persisting problems in the area of complex injuries of the extremities has to be pursued intensively.

Weight-bearing Restrictions after Distal Femur Fractures - Review of Current Literature. / Belastungsvorgaben nach distaler Femurfraktur - eine aktuelle Literaturübersicht.

The incidence of distal femur fractures increases in the geriatric patient. The primary treatment goal in these fractures is early mobilisation to prevent secondary injuries associated with immobility. In light of the increasing spectrum of therapeutic options for postoperative fracture treatment, including double plating, nail-plate combination and distal femur replacement as postoperative treatments, weight-bearing recommendations are becoming increasingly important. The aim of this study was thus to analyse the weight-bearing recommendations and associated therapy results within the literature of the past 9 years and compare the recommendations to our own approach.

[Primary joint replacement for the treatment of acetabular fractures]. / Primär endoprothetische Versorgung von Acetabulumfrakturen.

Open reduction and internal fixation are the gold standard for the treatment of dislocated acetabular fractures. A primary joint replacement is only justified in isolated cases. The indications are merely non-reconstructable acetabular fractures, accompanying displaced fractures of the femoral neck and non-reconstructable fractures of the femoral head.Because of the difficulties in achieving sufficient cup stability, joint replacement for the treatment of acetabular fractures regularly requires implants designed for revision arthroplasty. The Kocher-Langenbeck approach provides the most versatile options, as it enables simultaneous stabilization of the dorsal acetabular structures, which are essential for the stability of the cup.For primary joint replacement as a treatment of acetabular fractures, survival of the prosthetic cup is markedly worse when compared to elective primary joint replacement. Particularly in younger patients but also in aged patients, every effort should be made to achieve a press fit of the cup. In most cases, this will include reduction and fixation of the fracture for stabilization of the acetabulum prior to joint replacement. A staged approach with an early secondary replacement intervention seems to provide better overall results than simultaneous fracture fixation and joint replacement.

Photoacoustic imaging for the study of oxygen saturation and total hemoglobin in bone healing and non-union formation.

Non-union formation represents a major complication in trauma surgery. Adequate vascularization has been recognized as vital for bone healing. However, the role of vascularization in the pathophysiology of non-union formation remains elusive. This is due to difficulties in studying bone microcirculation in vivo. Therefore, we herein studied in a murine osteotomy model whether photoacoustic imaging may be used to analyze vascularization in bone healing and non-union formation. We found that oxygen saturation within the callus tissue is significantly lower in non-unions compared to unions and further declines over time. Moreover, the amount of total hemoglobin (HbT) within the callus tissue was markedly reduced in non-unions. Correlation analyses showed a strong positive correlation between microvessel density and HbT, indicating that photoacoustically determined HbT is a valid parameter to assess vascularization during bone healing. In summary, photoacoustic imaging is a promising approach to study vascular function and tissue oxygenation in bone regeneration.