Georg N. Duda

Internal forces and moments in the femur during walking

The forces exerted by the soft and hard tissues of the thigh together represent a system in equilibrium. This balance of loads must be considered when the system components are examined independently. However, in many biomechanical analyses of the thigh, the femur is studied without considering soft tissue loading. To improve the understanding of femoral loading a three-dimensional model was developed. Taking into account all thigh muscles, body weight and contact forces at the hip, patello-femoral and knee joints, the internal loads of the bone were calculated. Internal loads of the femur decreased as a result of muscle activity from proximal to distal at the hip and from distal to proximal at the knee. The load reduction could be up to 50% of the internal forces at the hip, depending on gait phase. Maximal forces were found between 40 and 60% of the stance phase, whereas maximal torsional moments occurred shortly after heel strike. This model demonstrated that muscles play a substantial role in balancing the loads within the femur. In general, the bone is loaded axially, rather than in bending, with maximum shear forces at the proximal and distal ends. Bending moments are relatively small compared to models which do not consider muscle activity. From one gait phase to another, the femur experiences alternating, rather than one-sided bending load.

Mesenchymal Stem Cells Regulate Angiogenesis According to Their Mechanical Environment

In fracture and bone defect healing, MSCs largely drive tissue regeneration. MSCs have been shown to promote angiogenesis both in vivo and in vitro. Angiogenesis is a prerequisite to large tissue reconstitution. The present study investigated how mechanical loading of MSCs influences their proangiogenic capacity. The results show a significant enhancement of angiogenesis by conditioned media from mechanically stimulated compared with unstimulated MSCs in two‐dimensional tube formation and three‐dimensional spheroid sprouting assays. In particular, proliferation but not migration or adhesion of endothelial cells was elevated. Promotion of angiogenesis was dependent upon fibroblast growth factor receptor 1 (FGFR1) signaling. Moreover, stimulation of tube formation was inhibited by vascular endothelial growth factor receptor (VEGFR) tyrosine kinase blocking. Screening for the expression levels of different soluble regulators of angiogenesis revealed an enrichment of matrix metalloprotease 2, transforming growth factor β1, and basic fibroblast growth factor but not of vascular endothelial growth factor in response to mechanical stimulation. In conclusion, mechanical loading of MSCs seems to result in a paracrine stimulation of angiogenesis, most likely by the regulation of a network of several angiogenic molecules. The underlying mechanism appears to be dependent on the FGFR and VEGFR signaling cascades and might be mediated by an additional cross‐talk with other pathways.

VARIABILITY OF FEMORAL MUSCLE ATTACHMENTS

Analytical and experimental models of the musculoskeletal system often assume single values rather than ranges for anatomical input parameters. The hypothesis of the present study was that anatomical variability significantly influences the results of biomechanical analyses, specifically regarding the moment arms of the various thigh muscles. Insertions and origins of muscles crossing or attaching to the femur were digitized in six specimens. Muscle volumes were measured; muscle attachment area and centroid location were computed. To demonstrate the influence of inter-individual anatomic variability on a mechanical modeling parameter, the corresponding range of muscle moment arms were calculated. Standard deviations, as a percentage of the mean, were about 70% for attachment area and 80% for muscle volume and attachment centroid location. The resulting moment arms of the m. gluteus maximus and m. rectus femoris were especially sensitive to anatomical variations (SD 65%). The results indicate that sensitivity to anatomical variations should be analyzed in any investigation simulating musculoskeletal interactions. To avoid misinterpretations, investigators should consider using several anatomical configurations rather than relying on a mean data set.

Realistic loads for testing hip implants

The aim here was to define realistic load conditions for hip implants, based on in vivo contact force measurements, and to see whether current ISO standards indeed simulate real loads. The load scenarios obtained are based on in vivo hip contact forces measured in 4 patients during different activities and on activity records from 31 patients. The load scenarios can be adapted to various test purposes by applying average or high peak loads, high-impact activities or additional low-impact activities, and by simulating normal or very active patients. The most strenuous activities are walking (average peak forces 1800 N, high peak forces 3900 N), going up stairs (average peak forces 1900 N, high peak forces 4200 N) and stumbling (high peak forces 11,000 N). Torsional moments are 50% higher for going up stairs than for walking. Ten million loading cycles simulate an implantation time of 3.9 years in active patients. The in vitro fatigue properties of cementless implant fixations are exceeded during stumbling. At least for heavyweight and very active subjects, the real load conditions are more critical than those defined by the ISO standards for fatigue tests.

The organization of the osteocyte network mirrors the extracellular matrix orientation in bone

Bone is a dynamic tissue that is continually undergoing a process of remodeling - an effect due to the interplay between bone resorption by osteoclasts and bone formation by osteoblasts. When new bone is deposited, some of the osteoblasts are embedded in the mineralizing collagen matrix and differentiate to osteocytes, forming a dense network throughout the whole bone tissue. Here, we investigate the extent to which the organization of the osteocyte network controls the collagen matrix arrangement found in various bone tissues. Several tissue types from equine, ovine and murine bone have been examined using confocal laser scanning microscopy as well as polarized light microscopy and back-scattered electron imaging. From comparing the spatial arrangements of unorganized and organized bone, we propose that the formation of a highly oriented collagen matrix requires an alignment of osteoblasts whereby a substrate layer provides a surface such that osteoblasts can align and, collectively, build new matrix. Without such a substrate, osteoblasts act isolated and only form matrices without long range order. Hence, we conclude that osteoblasts synthesize and utilize scaffold-like primary tissue as a guide for the deposition of highly ordered and mechanically competent bone tissue by a collective action of many cells.

Where should implants be anchored in the humeral head

To determine histomorphometric and bone strength distribution of the proximal humerus, analyses were done on 24 freshly harvested human cadaveric humeri. Median ages of 46 and 69 years were recorded respectively for the male group (n = 11; minimum, 34 years; maximum, 76 years) and the female group (n = 13; minimum, 46 years; maximum, 90 years). The humeral head was sliced into four equal horizontal levels (Levels 1–4). Five regions of interest were defined in each cutting plane: anterior, posterior, lateral, medial, and central. Histomorphometric analyses evaluated structural parameters (tissue volume to bone volume ratio, trabecular thickness), connectivity (number of nodes, node to node length), and trabecular orientation (mean bone length). The peak values of histomorphometric parameters and bone strength were identified for the cranial section and decreased caudally. The medial and dorsal aspects of the proximal humeral head were found to be the areas of highest bone strength. The trabecular network formed a pattern that connected the center of the gleaned cavity. The structural and connectivity parameters, bone strength, and trabecular orientation showed region- and level-related characteristics. Knowledge of distribution, microstructure, and quality of bone in the humeral head allows the remaining bone stock to be used effectively, even in elderly patients, with a minimally invasive approach and maximum mechanical stability.

Biomechanical Comparison of Cervical Spine Interbody Fusion Cages

Study Design. An in vitro biomechanical study of cervical spine interbody fusion cages using a sheep model was conducted. Objectives. To evaluate the biomechanical effects of cervical spine interbody fusion cages, and to compare three different cage design groups. Summary and Background Data. Recently, there has been a rapid increase in the use of cervical spine interbody fusion cages as an adjunct to spondylodesis. These cages can be classified into three design groups: screw, box, or cylinder designs. Although several comparative biomechanical studies of lumbar interbody fusion cages are available, biomechanical data for cervical spine constructs are lacking. Additionally, only limited data are available concerning comparative evaluation of different cage designs. Methods. In this study, 80 sheep cervical spines (C2–C5) were tested in flexion, extension, axial rotation, and lateral bending with a nondestructive stiffness method using a nonconstrained testing apparatus. Three-dimensional displacement was measured using an optical measurement system (Qualysis). Complete discectomy (C3–C4) was performed. Cervical spine interbody fusion cages were implanted according to manufacturers’ information. Eight spines in each of the the following groups were tested: intact, autologous iliac bone graft, two titanium screws (Novus CTTi; Sofamor Danek, Koln, Germany), two titanium screws (BAK-C 8 mm; Sulzer Orthopedics, Baar, Switzerland), one titanium screw (BAK-C 12 mm; Sulzer Orthopedics), carbon box (Novus CSRC; Sofamor Danek), titanium box (Syncage; Synthes, Bochum, Germany), titanium mesh cylinder (Harms; DePuy Acromed, Sulzbach, Germany), titanium cylinder (MSD; Ulrich, Ulm, Germany), and titanium cylinder (Kaden; BiometMerck, Berlin, Germany). The mean apparent stiffness values were calculated from the corresponding load-displacement curves. Additionally, cage volume and volume-related stiffness was determined. Results. After cervical spine interbody fusion cage implantation, flexion stiffness increased, as compared with that of the intact motion segment. On the contrary, rotation stiffness decreased after implantation of a cervical spine interbody fusion cage, except for the Novus CSRC, Syncage, and Kaden-Cage. If two screws were inserted (Novus CTTi and BAK-C 8 mm), there was no significant difference in flexion stiffness between screw and cylinder design groups. If one screw was inserted (BAK-C 12 mm), flexion stiffness was higher for cylinder designs (P < 0.05). Extension and bending stiffness were always higher with cylinder designs (P < 0.05). Volume-related stiffnessfor flexion extension and bending was highest for theHarms cage (P < 0.05). There was no difference for rotation volume-related stiffness between Harms and Syncage. Conclusions. The biomechanical results indicate that design variations in screw and cylinder design groups are of little importance. In this study, however, cages with a cylinder design were able to control extension and bending more effectively than cages with a screw design.

Composite transcriptome assembly of RNA-seq data in a sheep model for delayed bone healing

BackgroundThe sheep is an important model organism for many types of medically relevant research, but molecular genetic experiments in the sheep have been limited by the lack of knowledge about ovine gene sequences.ResultsPrior to our study, mRNA sequences for only 1,556 partial or complete ovine genes were publicly available. Therefore, we developed a composite de novo transcriptome assembly method for next-generation sequence data to combine known ovine mRNA and EST sequences, mRNA sequences from mouse and cow, and sequences assembled de novo from short read RNA-Seq data into a composite reference transcriptome, and identified transcripts from over 12 thousand previously undescribed ovine genes. Gene expression analysis based on these data revealed substantially different expression profiles in standard versus delayed bone healing in an ovine tibial osteotomy model. Hundreds of transcripts were differentially expressed between standard and delayed healing and between the time points of the standard and delayed healing groups. We used the sheep sequences to design quantitative RT-PCR assays with which we validated the differential expression of 26 genes that had been identified by RNA-seq analysis. A number of clusters of characteristic expression profiles could be identified, some of which showed striking differences between the standard and delayed healing groups. Gene Ontology (GO) analysis showed that the differentially expressed genes were enriched in terms including extracellular matrix, cartilage development, contractile fiber, and chemokine activity.ConclusionsOur results provide a first atlas of gene expression profiles and differentially expressed genes in standard and delayed bone healing in a large-animal model and provide a number of clues as to the shifts in gene expression that underlie delayed bone healing. In the course of our study, we identified transcripts of 13,987 ovine genes, including 12,431 genes for which no sequence information was previously available. This information will provide a basis for future molecular research involving the sheep as a model organism.

Joint line elevation in revision TKA leads to increased patellofemoral contact forces.

One difficulty in revision total knee arthroplasty (TKA) is the management of distal femoral bone defects in which a joint line elevation (JLE) is likely to occur. Although JLE has been associated with inferior clinical results, the effect that an elevated joint line has on knee contact forces has not been investigated. To understand the clinical observations and elaborate the potential risk associated with a JLE, we performed a virtual TKA on the musculoskeletal models of four subjects. Tibio‐ and patellofemoral joint contact forces (JCF) were calculated for walking and stair climbing, varying the location of the joint line. An elevation of the joint line primarily affected the patellofemoral joint with JCF increases of as much as 60% of the patients body weight (BW) at 10‐mm JLE and 90% BW at 15‐mm JLE, while the largest increase in tibiofemoral JCF was only 14% BW. This data demonstrates the importance of restoring the joint line, as it plays a critical role for the magnitudes of the JCFs, particularly for the patellofemoral joint. JLE caused by managing distal femoral defects with downsizing and proximalizing the femoral component could increase the patellofemoral contact forces, and may be a contributing factor to postoperative complications such as pain, polyethylene wear, and limited function.

Influence of age and mechanical stability on bone defect healing: Age reverses mechanical effects

Non-unions and delayed healing are still prevalent complications in fracture and bone defect healing. Both mechanical stability and age are known to influence this process. However, it remains unclear which factor dominates and how they interact. Within this study, we sought a link between both factors. In 36 female Sprague-Dawley rats, the left femur was osteotomized, distracted to an osteotomy gap of 1.5 mm and externally fixated. Variation of age (12 vs. 52 weeks - biologically challenging) and fixator stiffness (mechanically challenging) resulted in 4 groups (each 9 animals): YS: young semi-rigid, OS: old semi-rigid, YR: young rigid and OR: old rigid. Qualitative and quantitative radiographical analyses were performed at weeks 2, 4 and 6 after surgery. Six weeks post-op, rats were sacrificed and femora were harvested for biomechanical testing (torsional stiffness (TS) and maximum torque at failure (MTF)). Six weeks after surgery, TS showed a significant interaction between age and fixation stiffness (p<0.0001). TS in YR was significantly higher than that in the other groups (YS: p<0.001; OR: p<0.001; OS: p<0.001). Additionally, YS showed a significantly higher TS compared to the OS (p=0.006) and OR (p=0.046). Testing of MTF showed a significant interaction of both variables (p=0.0002) and led to significant differences between OR and YS (p<0.001), OS (p=0.046) and YR (p<0.001). The YR showed a higher MTF compared to YS (p=0.012) and OS (p=0.001), whereas ORs MTF was inferior compared to OS. At 2-week follow-up, YR (p=0.006), and at 6-week follow-up, YS and YR (p=0.032) showed significantly higher radiographic scores. At 2-week follow-up, YSs callus was larger than that of the old groups (OS: p=0.025; OR: p=0.003). In YR a significantly smaller callus was observed compared to YS at time points 4 and 6 weeks (p=0.002 for both) and compared to OS at 6-week follow-up (p=0.03). The effect of age seems to invert the effect of mechanical properties of the callus, which was not correlated to callus size. Optimization of mechanics alone seems to be not sufficient. The underlying mechanisms and causes of the age-related influences and their clinical counterparts need to be further investigated.