A new technique for internal fixation of femoral fractures in mice: impact of stability on fracture healing

Mouse models are of increasing interest to study the molecular aspects of fracture healing. Because biomechanical factors greatly influence the healing process, stable fixation of the fracture is of interest also in mouse models. Unlike in large animals, however, there is a lack of mouse models which provide stable osteosynthesis. The purpose of this study was therefore to develop a technique for a more stable fixation of femoral fractures in mice and to analyze the impact of stability on the process of fracture healing. The new technique introduced herein includes an intramedullary pin and an extramedullary metallic clip. Ex vivo biomechanical analysis revealed a significantly higher implant stiffness of our pin-clip technique when compared with previously described intramedullary fixation techniques. In vivo, we studied the course of healing after the more stable fixation with our pin-clip technique and compared the results with that observed after unstable fixation with the pin-clip technique after cutting the clip. After 2 and 5 weeks of fracture healing radiological analysis demonstrated that the more stable fixation with the pin-clip technique results in a significantly higher union rate compared to the unstable fixation. Torsional stiffness at 5 weeks was almost 3-fold of that measured after unstable fixation. Histomorphological analysis further showed that fractures stabilized with the pin-clip technique healed with a smaller periosteal callus area, an increased fraction of bone and a reduced amount of fibrous tissue. Of interest, the pin-clip fixation showed reliable union after 5 weeks, whereas the unstable pin fixation did not regularly achieve adequate fracture healing. In conclusion, we introduce a novel, easily applicable internal osteosynthesis technique in mice, which provides rotational stability after femoral fracture fixation. We further show that a more stable osteosynthesis significantly improves the process of fracture healing also in mice.     

Design and performance of a fracture monitoring transducer

The answer to the question, ‘when is a fracture healed?’ is not simple, since the healing process is progressive and it is not possible to specify a time when the fracture can be said to have healed. In the past the assessment of fracture healing has, in the main, been subjective, relying upon the skill of the interpreter. A more objective method would be an advantage for many reasons, and since the bone is intended to be load bearing it is reasonable to assess healing by measuring the mechanical integrity of the bone. to do this a ‘clamp on’ transducer has been developed which, when fitted to the support column of an external fixator, enables the stiffness of a fracture to be determined during the healing process. Over the past 6 years this system has been used for both clinical and research work. It has enabled various forms of treatment to be evaluated in terms of ‘rate of healing’ and it also indicates the safe point at which the fixator can be removed.     

Trabecular bone fracture healing simulation with finite element analysis and fuzzy logic

Trabecular bone fractures heal through intramembraneous ossification. This process differs from diaphyseal fracture healing in that the trabecular marrow provides a rich vascular supply to the healing bone, there is very little callus formation, woven bone forms directly without a cartilage intermediary, and the woven bone is remodelled to form trabecular bone. Previous studies have used numerical methods to simulate diaphyseal fracture healing or bone remodelling, however not trabecular fracture healing, which involves both tissue differentiation and trabecular formation. The objective of this study was to determine if intramembraneous bone formation and remodelling during trabecular bone fracture healing could be simulated using the same mechanobiological principles as those proposed for diaphyseal fracture healing. Using finite element analysis and the fuzzy logic for diaphyseal healing, the model simulated formation of woven bone in the fracture gap and subsequent remodelling of the bone to form trabecular bone. We also demonstrated that the trabecular structure is dependent on the applied loading conditions. A single model that can simulate bone healing and remodelling may prove to be a useful tool in predicting musculoskeletal tissue differentiation in different vascular and mechanical environments.     

Virtual structural analysis of tibial fracture healing from low-dose clinical CT scans

Quantitative assessment of bone fracture healing remains a significant challenge in orthopaedic trauma research. Accordingly, we developed a new technique for assessing bone healing using virtual mechano-structural analysis of computed tomography (CT) scans. CT scans from 19 fractured human tibiae at 12 weeks after surgery were segmented and prepared for finite element analysis (FEA). Boundary conditions were applied to the models to simulate a torsion test that is commonly used to access the structural integrity of long bones in animal models of fracture healing. The output of each model was the virtual torsional rigidity (VTR) of the healing zone, normalized to the torsional rigidity of each patient’s virtually reconstructed tibia. This provided a structural measure to track the percentage of healing each patient had undergone. Callus morphometric measurements were also collected from the CT scans. Results showed that at 12 weeks post-op, more than 75% of patients achieved a normalized VTR (torsional rigidity relative to uninjured bone) of 85% or above. The predicted intact torsional rigidities compared well with published cadaveric data. Across all patients, callus volume and density were weakly and non-significantly correlated with normalized VTR and time to clinical union. Conversely, normalized VTR was significantly correlated with time to union (R² = 0.383, p = 0.005). This suggests that fracture scoring methods based on the visual appearance of callus may not accurately predict mechanical integrity. The image-based structural analysis presented here may be a useful technique for assessment of bone healing in orthopaedic trauma research.     

Biomechanical assessment and clinical analysis of different intramedullary nailing systems for oblique fractures

The aim of this study is to evaluate the fracture union or non-union for a specific patient that presented oblique fractures in tibia and fibula, using a mechanistic-based bone healing model. Normally, this kind of fractures can be treated through an intramedullary nail using two possible configurations that depends on the mechanical stabilisation: static and dynamic. Both cases are simulated under different fracture geometries in order to understand the effect of the mechanical stabilisation on the fracture healing outcome. The results of both simulations are in good agreement with previous clinical experience. From the results, it is demonstrated that the dynamization of the fracture improves healing in comparison with a static or rigid fixation of the fracture. This work shows the versatility and potential of a mechanistic-based bone healing model to predict the final outcome (union, non-union, delayed union) of realistic 3D fractures where even more than one bone is involved.     

Dynamic analysis of a simplified bone model during the process of fracture healing

The dynamic analysis of fracture healing is tackled numerically by means of a bone model which uses the finite element method. The model is of non-uniform cross-sectional area and moment of inertia. Shear and rotatory inertia are taken into account. Considerable variation of the upper natural frequencies is observed as the healing process progresses. The practical implications, as well as present limitations, of the technique are examined.     

In vivo gait analysis in a mouse femur fracture model

Although the mouse has become a preferred species for molecular studies on fracture healing, gait analysis after fracture fixation and during bone healing has not yet been performed in mice. Herein, we introduce a novel technique for gait analysis in mice and report the change of motion pattern after fracture and fixation. A standardized femur fracture was stabilized by a common pin. The non-fractured tibia was additionally marked with a pin, allowing continuous analysis of the tibio-femoral angle by digital video-radiography. Dynamic gait analysis was performed at day fourteen after surgery in a radio-opaque running wheel. Fracture fixation resulted in a significantly reduced range and maximum of the tibio-femoral angle compared to non-fractured controls. This was associated with a significantly reduced stride length. Because stride frequency was slightly increased and, thus, stride time diminished, stride velocity was not significantly reduced compared to controls. Thus, our study demonstrates distinct alterations of the gait of mice at 2 weeks after femur fracture and stabilization. Our results support the need of gait analysis in fracture healing studies to assess the animals’ well-being.     

llizarov骨搬运技术治疗胫骨骨缺损的疗效及术后延迟愈合或不愈合的影响因素分析

摘要 目的：探讨 llizarov骨搬运技术治疗胫骨骨缺损的疗效及术后延迟愈合或不愈合的影响因素分析。方法：选取 2016年 6月-2020年 10月本院收治的 90例胫骨骨缺损患者为研究对象,患者均给予 llizarov骨搬运技术治疗。对患者的手术效果指标、并发症发生率进行记录统计。并对患者进行门诊随访观察,统计患者延迟愈合或不愈合的发生情况,据此将患者分为愈合组和延迟愈合或不愈合组。采用单因素及多因素 Logistic回归分析患者术后延迟愈合或不愈合发生的影响因素。结果：患者住院时间为(12.11± 2.98)d、开始负重时间为(45.39± 7.78)d、完全负重时间(76.41± 11.23)d。患者术后并发症发生率为 8.89%(8/90)。经随访观察,共有 29例患者出现术后延迟愈合或不愈合,发生率为 32.22%（29/90）。而延迟愈合或不愈合组患者的伤口感染、合并软组织损伤、合并腓骨骨折、术后过早活动及有吸烟史的人数占比高于愈合组患者(P<0.05)。经多因素 Logistic回归分析显示：伤口感染、合并软组织损伤、合并腓骨骨折、术后过早活动、有吸烟史是患者术后延迟愈合或不愈合的危险因素(P<0.05)。结论：llizarov骨搬运技术治疗胫骨骨缺损的疗效较好,患者的手术时间短、术中失血量少、住院时间、开始负重时间均较短,并发症发生率低,治疗安全性较好,但患者易出现术后延迟愈合或不愈合现象,可能与伤口感染、合并软组织损伤、合并腓骨骨折、术后过早活动、吸烟史有关。     

In vivo measurement of bending stiffness in fracture healing

Background  

Measurement of the bending stiffness a healing fracture represents a valid variable in the assessment of fracture healing. However, currently available methods typically have high measurement errors, even for mild pin loosening. Furthermore, these methods cannot provide actual values of bending stiffness, which precludes comparisons among individual fractures. Thus, even today, little information is available with regards to the fracture healing pattern with respect to actual values of bending stiffness. Our goals were, therefore: to develop a measurement device that would allow accurate and sensitive measurement of bending stiffness, even in the presence of mild pin loosening; to describe the course of healing in individual fractures; and help to evaluate whether the individual pattern of bending stiffness can be predicted at an early stage of healing.     

Developmental aspects of fracture healing and the use of pharmacological agents to alter healing

Fracture healing is a specialized postnatal repair process that recapitulates many aspects of embryological skeletal development. While many of the molecular mechanisms that control cellular differentiation and growth during embryogenesis recur during fracture healing, these processes take place in a postnatal environment that is unique and distinct from those which exist during embryogenesis. A number of the central biological processes that are believed to be crucial in the embryonic differentiation and growth of skeletal tissues and play a functional role in fracture healing are reviewed. The functional modification of these various developmental processes of fracture healing is discussed in the context of how different pharmacological agents might alter fracture healing.     

bFGF/BMP/胶原膜加速颌骨骨折愈合的大体及X线观察

目的：了解胶原膜作为生长因子缓释材料治疗颌骨骨折的应用前景。方法：将100μg的rhBMP-2用1ml的bFGF溶液完全溶解;用移液器移出40μl的该溶液,滴加到面积为0.5cm×1cm的胶原膜组织块中,冻干后制成生长因子缓释系统;在12只新西兰大白兔两侧制成人工下颌骨骨折模型,左侧置放bFGF/BMP/胶原膜;右侧均为空白对照;术后2、4、12周行临床大体观察及X线片观察。结果：实验组骨折愈合速度明显快于对照组。术后2周,X线结果显示bFGF/BMP/胶原膜组骨折断端边缘模糊。对照组骨折线明显。术后4周,X线结果显示bFGF/BMP/胶原膜骨折线基本不可见,骨折对位良好,断端边缘基本消失,骨折无错位。对照组骨折下缘可见纤维性骨痂形成,骨折线模糊。术后12周,各组X线结果无差异,骨折部位接近正常骨组织。结论：bFGF/BMP/胶原膜能加速骨折愈合,提高骨折愈合效果。     

Simulation of the nutrient supply in fracture healing

The healing process for bone fractures is sensitive to mechanical stability and blood supply at the fracture site. Most currently available mechanobiological algorithms of bone healing are based solely on mechanical stimuli, while the explicit analysis of revascularization and its influences on the healing process have not been thoroughly investigated in the literature. In this paper, revascularization was described by two separate processes: angiogenesis and nutrition supply. The mathematical models for angiogenesis and nutrition supply have been proposed and integrated into an existing fuzzy algorithm of fracture healing. The computational algorithm of fracture healing, consisting of stress analysis, analyses of angiogenesis and nutrient supply, and tissue differentiation, has been tested on and compared with animal experimental results published previously. The simulation results showed that, for a small and medium-sized fracture gap, the nutrient supply is sufficient for bone healing, for a large fracture gap, non-union may be induced either by deficient nutrient supply or inadequate mechanical conditions. The comparisons with experimental results demonstrated that the improved computational algorithm is able to simulate a broad spectrum of fracture healing cases and to predict and explain delayed unions and non-union induced by large gap sizes and different mechanical conditions. The new algorithm will allow the simulation of more realistic clinical fracture healing cases with various fracture gaps and geometries and may be helpful to optimise implants and methods for fracture fixation.     

Identification of tissue differentiation rates in a mechanobiological model of fracture healing

As a basis for model-based analysis of the processes in secondary fracture healing, a dynamical model is presented that characterises the physiological status in the fracture area by the location-dependent composition of tissues. Five types of tissue are distinguished: connective tissue, cartilage, bone, haematoma and avascular bone. A rule base is given that describes dynamical tissue differentiation processes. The rules consider not only a mechanical stimulus but also osteogenic and a vasculative factors as biological stimuli. Within this model structure, it is possible, e.g., to distinguish intramembranous from endochondral ossification processes. An objective function is introduced to assess accordance between the model-based simulation results and reference healing stages. By minimising this objective function, relevant tissue differentiation rates can be determined. For a reference process of secondary fracture healing it could be shown that the intramembranous ossification rate of 0.313%/day (from connective tissue to bone) is much smaller than the endochondral ossification rate of 1.136%/day (from cartilage to bone). In order to verify the model approach, it is transferred to simulate long bone distraction. Results show that healing patterns of bone distraction can be predicted. Using this method, it is possible to identify model parameters for individual subjects. This will allow a patient-specific analysis of tissue healing processes in future.     

A finite element inverse analysis to assess functional improvement during the fracture healing process

Assessment of the restoration of load-bearing function is the central goal in the study of fracture healing process. During the fracture healing, two critical aspects affect its analysis: (1) material properties of the callus components, and (2) the spatio-temporal architecture of the callus with respect to cartilage and new bone formation. In this study, an inverse problem methodology is used which takes into account both features and yields material property estimates that can analyze the healing changes. Six stabilized fractured mouse tibias are obtained at two time points during the most active phase of the healing process, respectively 10 days (n=3), and 14 days (n=3) after fracture. Under the same displacement conditions, the inverse procedure estimations of the callus material properties are generated and compared to other fracture healing metrics. The FEA estimated property is the only metric shown to be statistically significant (p=0.0194) in detecting the changes in the stiffness that occur during the healing time points. In addition, simulation studies regarding sensitivity to initial guess and noise are presented; as well as the influence of callus architecture on the FEA estimated material property metric. The finite element model inverse analysis developed can be used to determine the effects of genetics or therapeutic manipulations on fracture healing in rodents.     

Measurement of fracture movement in patients treated with unilateral external skeletal fixation

Axial movement occurring at the fracture site has been determined in a group of healing tibial fractures treated by external skeletal fixation. Fracture movement was determined via a strain gauge transducer which was attached to the column of the external fixator and measured the deflection of the bone screw adjacent to the fracture site and the active loading or weight bearing given by the patient to the fractured limb was monitored using a force platform. The results for 27 subjects show that, with a rigid unilateral fixator, the axial movement occurring at the fracture site was initially small (mean = 0.28 mm at 5 weeks post fracture). This movement increases to reach a mean maximum value of 0.43 mm at 11 weeks post-fracture and then decreases, despite increased weight bearing, as fracture healing progresses. In the early stages of healing, the movement can be increased slightly if the fixator is fitted with a module which permits additional fracture site movement, although the resultant increase in movement is only a small proportion of the potential available with this module.     

A hybrid bioregulatory model of angiogenesis during bone fracture healing

Bone fracture healing is a complex process in which angiogenesis or the development of a blood vessel network plays a crucial role. In this paper, a mathematical model is presented that simulates the biological aspects of fracture healing including the formation of individual blood vessels. The model consists of partial differential equations, several of which describe the evolution in density of the most important cell types, growth factors, tissues and nutrients. The other equations determine the growth of blood vessels as a result of the movement of leading endothelial (tip) cells. Branching and anastomoses are accounted for in the model. The model is applied to a normal fracture healing case and subjected to a sensitivity analysis. The spatiotemporal evolution of soft tissues and bone, as well as the development of a blood vessel network are corroborated by comparison with experimental data. Moreover, this study shows that the proposed mathematical framework can be a useful tool in the research of impaired healing and the design of treatment strategies.     

Callus Formation and Mineralization after Fracture with Different Fixation Techniques: Minimally Invasive Plate Osteosynthesis versus Open Reduction Internal Fixation

Minimally invasive plate osteosynthesis(MIPO) has been considered as an alternative for fracture treatment. Previous study has demonstrated that MIPO technique has the advantage of less soft tissue injury compared with open reduction internal fixation (ORIF). However, the comparison of callus formation and mineralization between two plate osteosynthesis methods remains unknown. In this experiment, ulna fracture model was established in 42 beagle dogs. The fractures underwent reduction and internal fixation with MIPO or ORIF. Sequential fluorescent labeling and radiographs were applied to determine new callus formation and mineralization in two groups after operation. At 4, 8 and 12 weeks postoperatively, the animals were selected to be sacrificed and the ulna specimens were analyzed by Micro-CT. The sections were also treated with Masson staining for histological evaluation. More callus formation was observed in MIPO group in early stage of fracture healing. The fracture union rate has no significant difference between two groups. The results indicate that excessive soft tissue stripping may impact early callus formation. As MIPO technique can effectively reduce soft tissue injury with little incision, it is considered to be a promising alternative for fracture fixation.     

Numerical Simulation of Callus Healing for Optimization of Fracture Fixation Stiffness

The stiffness of fracture fixation devices together with musculoskeletal loading defines the mechanical environment within a long bone fracture, and can be quantified by the interfragmentary movement. In vivo results suggested that this can have acceleratory or inhibitory influences, depending on direction and magnitude of motion, indicating that some complications in fracture treatment could be avoided by optimizing the fixation stiffness. However, general statements are difficult to make due to the limited number of experimental findings. The aim of this study was therefore to numerically investigate healing outcomes under various combinations of shear and axial fixation stiffness, and to detect the optimal configuration. A calibrated and established numerical model was used to predict fracture healing for numerous combinations of axial and shear fixation stiffness under physiological, superimposed, axial compressive and translational shear loading in sheep. Characteristic maps of healing outcome versus fixation stiffness (axial and shear) were created. The results suggest that delayed healing of 3 mm transversal fracture gaps will occur for highly flexible or very rigid axial fixation, which was corroborated by in vivo findings. The optimal fixation stiffness for ovine long bone fractures was predicted to be 1000–2500 N/mm in the axial and >300 N/mm in the shear direction. In summary, an optimized, moderate axial stiffness together with certain shear stiffness enhances fracture healing processes. The negative influence of one improper stiffness can be compensated by adjustment of the stiffness in the other direction.