Jenni M. Buckley

Role of Matrix Metalloproteinase 13 in Both Endochondral and Intramembranous Ossification during Skeletal Regeneration

Extracellular matrix (ECM) remodeling is important during bone development and repair. Because matrix metalloproteinase 13 (MMP13, collagenase-3) plays a role in long bone development, we have examined its role during adult skeletal repair. In this study we find that MMP13 is expressed by hypertrophic chondrocytes and osteoblasts in the fracture callus. We demonstrate that MMP13 is required for proper resorption of hypertrophic cartilage and for normal bone remodeling during non-stabilized fracture healing, which occurs via endochondral ossification. However, no difference in callus strength was detected in the absence of MMP13. Transplant of wild-type bone marrow, which reconstitutes cells only of the hematopoietic lineage, did not rescue the endochondral repair defect, indicating that impaired healing in Mmp13−/− mice is intrinsic to cartilage and bone. Mmp13−/− mice also exhibited altered bone remodeling during healing of stabilized fractures and cortical defects via intramembranous ossification. This indicates that the bone phenotype occurs independently from the cartilage phenotype. Taken together, our findings demonstrate that MMP13 is involved in normal remodeling of bone and cartilage during adult skeletal repair, and that MMP13 may act directly in the initial stages of ECM degradation in these tissues prior to invasion of blood vessels and osteoclasts.

Limitations of Standard Fluoroscopy in Detecting Rotational Malreduction of the Syndesmosis in an Ankle Fracture Model

Background: When treating ankle fractures with associated syndesmosis injury, failure to anatomically reduce the syndesmosis may lead to poor outcome. While shortening and posterior subluxation of the distal fibula are readily detected by intraoperative fluoroscopy, it is unclear how well malrotation can be assessed. The ability of fluoroscopy to detect rotational malre-duction of the fibula was the subject of this study. Materials and Methods: Distal fibula fractures with complete syndesmotic injury were produced in ten cadaveric ankles. Two Kirschner wires were used to fix the fibula in neutral (0 degrees), 10 to 30 degrees of external rotation (ER), and 10 degrees to 30 degrees of internal rotation (IR). Using C-arm fluoroscopy tibio-fibular clear space and tibio-fibular overlap in the AP and mortise views, and posterior fibular subluxation in the lateral view were measured to assess reduction of the syndesmosis. Results: The radiographic indices were able to detect as little as 10 degrees of IR but were within their normal range in up to 30 degrees of ER. When assessing for a 2mm difference compared to the intact ankle, sensitivity of all indices were low after more than 15 degrees ER, but high and clinically useful after more than 15 degrees of IR. Conclusion: Radiographic indices for syndesmosis disruption could not detect ER malreduction of the syndesmosis of up to 30 degrees. Clinical Relevance: In the setting of ankle fractures with syndesmosis disruption, fixing the fibula in as much as 30 degrees of external rotation may go undetected using intraoperative fluoroscopy alone.

The tissue diagnostic instrument

Tissue mechanical properties reflect extracellular matrix composition and organization, and as such, their changes can be a signature of disease. Examples of such diseases include intervertebral disk degeneration, cancer, atherosclerosis, osteoarthritis, osteoporosis, and tooth decay. Here we introduce the tissue diagnostic instrument (TDI), a device designed to probe the mechanical properties of normal and diseased soft and hard tissues not only in the laboratory but also in patients. The TDI can distinguish between the nucleus and the annulus of spinal disks, between young and degenerated cartilage, and between normal and cancerous mammary glands. It can quantify the elastic modulus and hardness of the wet dentin left in a cavity after excavation. It can perform an indentation test of bone tissue, quantifying the indentation depth increase and other mechanical parameters. With local anesthesia and disposable, sterile, probe assemblies, there has been neither pain nor complications in tests on patients. We anticipate that this unique device will facilitate research on many tissue systems in living organisms, including plants, leading to new insights into disease mechanisms and methods for their early detection.

The Fatigue Life of Contoured Cobalt Chrome Posterior Spinal Fusion Rods

Intraoperative contouring of posterior rods in lumbar arthrodesis constructs introduces stress concentrations that can substantially reduce fatigue life. The sensitivity of titanium (Ti) and stainless steel (SS) to intraoperative contouring has been established in the literature; however, notch sensitivity has yet to be quantified for cobalt chrome (CoCr), which is now being advocated for use in posterior arthrodesis constructs. The goal of this study is to evaluate the sensitivity of CoCr rods to intraoperative contouring for posterior lumbar screw—rod arthrodesis constructs. In this paper lumbar bilateral vertebrectomy models are constructed based on ASTM F1717-01 with curved rods (26—30° total curvature) and poly-axial pedicle screws. Three types of constructs are assembled: first, 5.5 mm SS rods with SS screws (6.5×35 mm), second, 6.0 mm Ti rods with Ti screws (7.5×35 mm), and third, 6.0 mm CoCr rods with Ti screws (7.5×35 mm). All specimens are tested at 4 Hz in dynamic axial compression-bending with a load ratio of ten and maximum load levels of 250, 400, and 700 N until run-out at 2 000 000 cycles. Results are presented that show that the fatigue life of CoCr constructs tend to be greater than Ti constructs at all levels. At the 400 N maximum loading, CoCr lasts an average of 350 000 cycles longer than the Ti constructs. The CoCr constructs are able to sustain the 250 N load until run-out at 2 000 000 cycles but they fail at high load levels (maximum 700 N). The CoCr constructs fail at the neck of the Ti screw at high loads whereas Ti screws fail at the notch induced by contouring. Since CoCr is compatible with magnetic resonance imaging and has high static strength characteristics, the results of this study suggest that it may be an appropriate substitute for Ti.

Quantitative computed tomography-based predictions of vertebral strength in anterior bending.

Study Design. This study examined the ability of QCT-based structural assessment techniques to predict vertebral strength in anterior bending. Objective. The purpose of this study was to compare the abilities of QCT-based bone mineral density (BMD), mechanics of solids models (MOS), e.g., bending rigidity, and finite element analyses (FE) to predict the strength of isolated vertebral bodies under anterior bending boundary conditions. Summary of Background Data. Although the relative performance of QCT-based structural measures is well established for uniform compression, the ability of these techniques to predict vertebral strength under nonuniform loading conditions has not yet been established. Methods. Thirty human thoracic vertebrae from 30 donors (T9–T10, 20 female, 10 male; 87 ± 5 years of age) were QCT scanned and destructively tested in anterior bending using an industrial robot arm. The QCT scans were processed to generate specimen-specific FE models as well as trabecular bone mineral density (tBMD), integral bone mineral density (iBMD), and MOS measures, such as axial and bending rigidities. Results. Vertebral strength in anterior bending was poorly to moderately predicted by QCT-based BMD and MOS measures (R2 = 0.14–0.22). QCT-based FE models were better strength predictors (R2 = 0.34–0.40); however, their predictive performance was not statistically different from MOS bending rigidity (P > 0.05). Conclusions. Our results suggest that the poor clinical performance of noninvasive structural measures may be due to their inability to predict vertebral strength under bending loads. While their performance was not statistically better than MOS bending rigidities, QCT-based FE models were moderate predictors of both compressive and bending loads at failure, suggesting that this technique has the potential for strength prediction under nonuniform loads. The current FE modeling strategy is insufficient, however, and significant modifications must be made to better mimic whole bone elastic and inelastic material behavior.

Effect of severity of rod contour on posterior rod failure in the setting of lumbar pedicle subtraction osteotomy (PSO): a biomechanical study.

BACKGROUND Rod failure has been reported clinically in pedicle subtraction osteotomy (PSO) to correct flat back deformity. OBJECTIVE To characterize the fatigue life of posterior screw-rod constructs in the setting of PSO as a function of the severity of rod contour angle. METHODS A modified ASTM F1717 to 04 was used. Rods were contoured to the appropriate angle for the equivalent 20-, 40-, or 60-degree PSO angles. Testing was performed on a mechanical test frame at 400/40 N and 250/25 N, and specimens were cycled at 4 Hz to failure or run-out at 2,000,000 cycles. The effect of the screw-rod system on fatigue strength of curved rods was compared using Cox proportional hazards regression. RESULTS At 400 N/40 N, Cox proportional hazards regression indicated that contouring rods from a 20-degree PSO angle to either 40 or 60 degrees significantly decreased fatigue life (hazard ratio = 7863.6, P = .0144). However, contouring rods from a 40-degree PSO angle to 60 degrees had no significant effect on the fatigue life (P > .05). At 250 N/25 N, Cox proportional hazards regression indicated that contouring rods from a 20-degree PSO angle to either 40 or 60 degrees significantly decreased fatigue life (hazard ratio = 7863.6, P = .0144). Furthermore, contouring rods from a 40-degree PSO angle to 60 degrees had a significant effect on the fatigue life (hazard ratio = 7863.6, P = .0144). CONCLUSION Results suggest that in the setting of PSO, the fatigue life of posterior spinal fixation rods depends largely on the severity of the rod angle used to maintain the vertebral angle created by the PSO and is significantly lowered by rod contouring.

Biomechanical comparison of three fixation techniques for unstable thoracolumbar burst fractures. Laboratory investigation.

OBJECT Increased structural stability is considered sufficient justification for higher-risk surgical procedures, such as circumferential fixation after severe spinal destabilization. However, there is little biomechanical evidence to support such claims, particularly after traumatic lumbar burst fracture. The authors sought out to compare the biomechanical performance of the following 3 fixation strategies for spinal reconstruction after decompression for an unstable thoracolumbar burst fracture: 1) short-segment anterolateral fixation; 2) circumferential fixation; and 3) extended anterolateral fixation. METHODS Thoracolumbar spines (T10-L4) from 7 donors (mean age at death 64+/-6 years; 1 female and 6 males) were tested in pure moment loading in flexion-extension, lateral bending, and axial rotation. Thoracolumbar burst fractures were surgically induced at L-1, and testing was repeated sequentially for each of the following fixation techniques: short-segment anterolateral, circumferential, and extended anterolateral. Primary and coupled 3D motions were measured across the instrumented site (T12-L2) and compared across treatment groups. RESULTS Circumferential and extended anterolateral fixations were statistically equivalent for primary and off-axis range-of-motions in all loading directions, and short-segment anterolateral fixation offered significantly less rigidity than the other 2 methods. CONCLUSIONS The results of this study strongly suggest that extended anterolateral fixation is biomechanically comparable to circumferential fusion in the treatment of unstable thoracolumbar burst fractures with posterior column and posterior ligamentous injury. In cases in which an anterior procedure may be favored for load sharing or canal decompression, extension of the anterior instrumentation and fusion one level above and below the unstable segment can result in near equivalent stability to a 2-stage circumferential procedure.

Tibial plateau fracture repairs augmented with calcium phosphate cement have higher in situ fatigue strength than those with autograft.

Objectives: This study compared the biomechanical fatigue strength of calcium phosphate augmented repairs versus autogenous bone graft (ABG) repairs in lateral tibia plateau fractures. Methods: Eight matched pairs of tibias (six male, two female; age, 75 ± 14 years) were harvested from fresh-frozen cadavers. Reproducible split-depression fractures were simulated and repaired by an orthopaedic traumatologist using a lateral tibial plateau plate. One tibia from each donor was randomly assigned to either calcium phosphate (Callos; Acumed, Hillsboro, OR) or ABG as augmentation. The femoral component of a hemitotal knee arthroplasty was attached to the actuator of a servohydraulic press and centered above the repair site. Cyclic, physiological compression loads were applied at 4Hz starting with a maximum load of 15% body weight and increasing by 15% body weight every 70,000 cycles. Loading conditions were determined from calculations of weight distribution, joint contact area, and gait characterization from existing literature. Repair site depression and stiffness were measured at regular intervals. Specimens were then loaded to failure at 1 mm/min. Results: Calcium phosphate augmented repairs subsided less and were more stiff during the fatigue loading than were ABG repairs at the 70,000, 140,000, and 210,000 cycle intervals (P < 0.03) All repairs survived to 210,000 cycles. The average ultimate load of the calcium phosphate repairs was 2241 ± 455 N (N = 6) and 1717 ± 508 N (N = 8) for ABG repairs (P = 0.02). Conclusion: Calcium phosphate repairs have significantly higher fatigue strength and ultimate load than ABG repairs and may increase the immediate weightbearing capabilities of the repaired knee.

Inter-laboratory variability in in vitro spinal segment flexibility testing

In vitro spine flexibility testing has been performed using a variety of laboratory-specific loading apparatuses and conditions, making test results across laboratories difficult to compare. The application of pure moments has been well established for spine flexibility testing, but to our knowledge there have been no attempts to quantify differences in range of motion (ROM) resulting from laboratory-specific loading apparatuses. Seven fresh-frozen lumbar cadaveric motion segments were tested intact at four independent laboratories. Unconstrained pure moments of 7.5 Nm were applied in each anatomic plane without an axial preload. At laboratories A and B, pure moments were applied using hydraulically actuated spinal loading fixtures with either a passive (A) or controlled (B) XY table. At laboratories C and D, pure moments were applied using a sliding (C) or fixed ring (D) cable-pulley system with a servohydraulic test frame. Three sinusoidal load-unload cycles were applied at laboratories A and B while a single quasistatic cycle was applied in 1.5 Nm increments at laboratories C and D. Non-contact motion measurement systems were used to quantify ROM. In all test directions, the ROM variability among donors was greater than single-donor ROM variability among laboratories. The maximum difference in average ROM between any two laboratories was 1.5° in flexion-extension, 1.3° in lateral bending and 1.1° in axial torsion. This was the first study to quantify ROM in a single group of spinal motion segments at four independent laboratories with varying pure moment systems. These data support our hypothesis that given a well-described test method, independent laboratories can produce similar biomechanical outcomes.

Biomechanical Pullout Strength and Stability of the Cervical Artificial Pedicle Screw

Study Design. This study used cadaveric specimens to compare the biomechanical performance of artificial pedicle screws (APS) versus lateral mass screws (LMS). Objective. The goal of this study was to biomechanically characterize APS range-of-motion and pullout strength in surgical instances that preclude LMS insertion. Summary of Background Data. Posterior approaches used in instances of ventral spine tumors often necessitate complete facetectomy, thereby removing fixation points for LMS and requiring longer constructs with fewer segmental fixations to span the resected levels. Recently, APS were developed to overcome this obstacle. Although APS have been used successfully in clinical cases, they have yet to be biomechanically validated. Methods. Seven fresh-frozen cervical spine segments (C2–C7) were harvested from human donors (F = 1, M = 6; 65 ± 5 years old, range: 50–72 years old). Nondestructive range-of-motion tests were conducted on each specimen in its intact and surgically destabilized states, and after each of 3 different APS and LMS surgical stabilizations. After nondestructive bending tests, a final pullout test of APS and LMS at the C4 level was performed for each specimen. Results. The pullout strength of the APS was twice as strong on average as that for the LMS (503.4 ± 338.3 vs.— 254.3 ± 142.3 N); this difference approached but was not statistically significant (P = 0.07). There was no significant difference in specimen stiffness between the APS- and LMS-instrumented configurations in all ranges of primary and off-axis motions (P > 0.05). However, all fixation methods increased specimen stiffness in comparison with the intact conditions (1.7–36.5 times increase; P < 0.05). Conclusion. Our results demonstrate that APS provide comparable stability to LMS and can therefore be considered a viable alternative in surgical scenarios requiring the complete removal of lateral masses. Moreover, APS may provide some enhanced strength in the face of destructive pullout forces.