Biomechanical Stiffness Research Articles

Natural Bone Collagen Scaffold Combined with Autologous Enriched Bone Marrow Cells for Induction of Osteogenesis in an Ovine Spinal Fusion Model

Autologous bone graft, the standard of bone grafting in achieving spinal fusion, is associated with several limitations and complications. The use of bone marrow cells (BMCs) as a potential cell source for spinal fusion, combined with a suitable scaffold to promote bone formation, may be a better choice. The aims of this study were to evaluate the efficacy of natural bone collagen scaffold (NBCS) combined with autologous-enriched BMCs for induction of osteogenesis in vitro and in vivo. Ovine-enriched BMCs were co-cultured with NBCS for 1, 2, 3, and 4 weeks to investigate whether NBCS would support the population expansion and differentiation of enriched BMCs. Using an ovine interbody fusion model, NBCS seeded with autologous enriched BMCs was implanted into the lumbar disc space. Fusion outcomes were compared with the use of the autograft, NBCS without BMCs, and BMCs without NBCS. In vitro results demonstrated that NBCS facilitated the population expansion and differentiation of ovine-enriched BMCs, promoting the expression of collagen type I and the formation of a mineralized matrix. The use of NBCS combined with enriched BMCs in vivo enhanced the spinal fusion rate (6 of 6 at 10 week) (p < 0.05), the biomechanical stiffness of fusion masses, and bone volume at the fusion site (p < 0.05). Histological findings also revealed that a combination of NBCS and BMCs induced new bone formation that integrated well with host bone tissue. In conclusion, NBCS is an effective scaffold that supports ovine-enriched BMCs. The combination of NBCS and BMCs may be a useful alternative for autograft in induction of spinal fusion.

Three-Dimensional In Vitro Effects of Compression and Time in Culture on Aggregate Modulus and on Gene Expression and Protein Content of Collagen Type II in Murine Chondrocytes

The objectives of this study were to determine how culture time and dynamic compression, applied to murine chondrocyte-agarose constructs, influence construct stiffness, expression of col2 and type II collagen. Chondrocytes were harvested from the ribs of six newborn double transgenic mice carrying transgenes that use enhanced cyan fluorescent protein (ECFP) and green fluorescent protein (GFP-T) as reporters for expression from the col2a1 and col1a1 promoters, respectively. Sixty-three constructs (8 mm diameter x 3 mm thick) per animal were created by seeding chondrocytes (10 x 10(6) per mL) in agarose gel (2% w/v). Twenty-eight constructs from each animal were stimulated for 7, 14, 21, or 28 days in a custom bioreactor housed in an electromagnetic system. Twenty-eight constructs exposed to identical culture conditions but without mechanical stimulation served as nonstimulated controls for 7, 14, 21, and 28 days. The remaining seven constructs served as day 0 controls. Fluorescing cells with rounded morphology were present in all constructs at all five time points. Seven, 14, 21, and 28 days of stimulation significantly increased col2 expression according to ECFP fluorescence and messenger RNA expression according to quantitative reverse transcriptase polymerase chain reaction. Col2 gene expression in stimulated and nonstimulated constructs showed initial increases up to day 14 and then showed decreases by day 28. Stimulation significantly increased type II collagen content at 21 and 28 days and aggregate modulus only at 28 days. There was a significant increase in aggregate modulus in stimulated constructs between day 0 and 7 and between day 21 and day 28. This study reveals that compressive mechanical stimulation is a potent stimulator of col2 gene expression that leads to measurable but delayed increases in protein (type II collagen) and then biomechanical stiffness. Future studies will examine the effects of components of the mechanical signal in culture and address the question of whether such in vitro improvements in tissue-engineered constructs enhance repair outcomes after surgery.

Transplantation of human mesenchymal stem cells in a non-autogenous setting for bone regeneration in a rabbit critical-size defect model

Human mesenchymal stem cells (hMSC) represent an attractive cell population for tissue engineering purposes. Furthermore, hMSC are described as immune privileged, and non-autogenous application seems possible. The current study examines the regeneration potential of hMSC after xenogenic transplantation compared with autogenous rabbit MSC in a critical-size bone defect. After isolation, hMSC and rabbit MSC were seeded on calcium-deficient hydroxyapatite (CDHA) and transplanted into a radial critical-size defect of New Zealand white rabbits. Defects were filled with a CDHA scaffold seeded with autogenous rabbit MSC, CDHA seeded with xenogenic hMSC or unseeded CDHA. An empty defect served as control group. Animals were sacrificed after 3 months. Evaluation was performed using radiography, micro-computed tomography (micro-CT) and histology. In addition, a non-destructive four-point-bending test was performed in order to evaluate biomechanical stiffness. While autogenous MSC seeded on CDHA led to increased healing of critical-size bone defects from radiological (micro-CT; p = 0.009) and histological (p = 0.048) perspectives compared with unloaded CDHA, it was not possible to demonstrate analogous effects for the xenogenic transplantation of hMSC. The xenogenic treatment group displayed inferior results in all parameters compared with the autogenous MSC treatment group (histology p = 0.041; micro-CT p = 0.006; biomechanical testing p = 0.017). Nevertheless, no local or systemic inflammatory response resulting from xenogenic transplantation was observed. While previous papers suggest the use of non-autogenous hMSC cells for tissue engineering purposes, the present results show inferior clinical results from transplantation of hMSC in a xenogenic setting compared with autogenous MSC.

Toward Regenerating a Human ThumbIn Situ

Regenerative technology promises to alleviate the problem of limited donor supply for bone or organ transplants. Most expensive and time consuming is cell expansion in laboratories. We propose a method of magnetically enriched osteoprogenitor stem cells, dispersed in self-assembling hydrogels and applied onto new ultra-high resolution, jet-based, three-dimensional printing of living human bone in a single-step for in situ bone regeneration. Human bone marrow-derived mesenchymal stem cells (hBMSCs) were enriched with CD 117+ cells, dispersed in different collagen I and RAD 16I hydrogel mixes, and applied onto three-dimensional printed btricalcium phosphate=poly(lactic-co-glycolic acid) scaffolds, printed from ultra-high-resolution volumetric CT images of a human thumb. Constructs were directly implanted subcutaneously into nude mice for 6 weeks. In vivo radiographic volumetric CT scanning and histological evaluations were performed at 1, 2, 4, and 6 weeks, and expression of bone-specific genes and biomechanical compression testing at 6 weeks endpoint. Time-dependant accumulation of bone-like extracellular matrix was most evident in CD 117+ hBMSCs using collagen I=RAD 16I hydrogel mix. This was shown histologically by Toluidine blue, von Kossa, and alkaline phosphatase staining, paralleled by increased radiological densities within implants approximating that of human bone, and confirmed by high expression of bone-specific osteonectin and biomechanical stiffness at 6 weeks. Human origin of newly formed tissue was established by expression of human GAPDH using RT-PCR. Statistical analysis confirmed high correlations between biomechanical stiffness, radiological densities, and bone markers. Bone tissue can be successfully regenerated in vivo using a single-step procedure with constructs composed of RAD 16I=collagen I hydrogel, CD 117+-enriched hBMSCs, and porous b-tricalcium phosphate=poly(lactic-co-glycolic acid) scaffolds.

Crosslinking by riboflavin and UVA. An electron transmission microscopy study in rabbits. Preliminary results

Abstract Purpose Collagen crosslinking of the cornea (CXL) has been developed recently as a promising treatment of keratoconus. In vitro measurements have demonstrated a significant increase in biomechanical stiffness of the crosslinked cornea. The aim of the present study was to evaluate the effect of this procedure on the collagen fiber diameter of the rabbit cornea over time. Methods The corneas of 10 New Zealand White albino rabbits were crosslinked by application of the photosensitizer riboflavin and exposure to UVA light (365 nm, 3 mW/cm2) for 30 minutes. The fellow control eyes were left untreated. On ultrathin sections of samples from the central and peripheral cornea, the collagen fiber diameter were measured with the help of morphometric computer software. Results In the anterior stroma, 48 hours after the CXL the collagen fiber diameter in the treated corneas was significantly increased by 10% compared with the control fellow eyes. Four months after the CXL, the mean collagen fiber diameters were decreased as compared to the 48 h specimens. In the 6 month specimens the diameter of the collagen fibers was further decreased as compared to the control fellow eyes up to ‐8%. Conclusion CXL leads to a significant increase in corneal collagen diameter 48 hours after the procedure. Interestingly, 4 months after the CXL, there is a decrease in the mean diameter of the collagen fiber and at 6 months, there is a further decrease up to 8% compared to the fellow eye. We can assume that the rigidity and the biomechanical stability of the cornea after CXL is increased because of the new intercollagen connections and collagen fiber diameter has little or no role in this procedure.

Role of Corneal Biomechanical Properties in Applanation Tonometry Measurements

To demonstrate the importance of material properties of the cornea in intraocular pressure (IOP) readings via standard Goldmann applanation tonometry. A realistic finite element model of the cornea was developed for the simulation of Goldmann applanation tonometry. A virtual cornea population was generated by randomly sampling material properties, central corneal thickness (CCT), and IOP for comparison with 181 clinical cases. The effect of material properties and CCT on IOP prediction in the virtual population was determined via computational simulation. The results show that corneal biomechanical properties (as characterized in this study by the stiffness parameter Einit) are as important as the CCT in influencing measured (Goldmann) IOP. This study supports the contention that the observed large scatter in standard correlations of clinical measurements of IOP versus CCT can be largely accounted for by plausible individual variations in corneal biomechanical stiffness properties.

Long‐term biomechanical properties of rabbit cornea after photodynamic collagen crosslinking

Photodynamic riboflavin/ultraviolet-A (UVA)-induced collagen cross-linking, which increases the biomechanical stiffness of the human cornea by about 300%, has been introduced recently as a possible treatment for progressive keratoconus. The present study was undertaken to evaluate the longterm biomechanical effects of this new cross-linking treatment as a necessary prerequisite to its clinical success. The corneas of the left eyes of nine male rabbits were cross-linked. The contralateral eyes served as controls. After removal of the central 7 mm of the epithelium, the corneas were treated with the photosensitizer riboflavin and UVA irradiation for 30 mins with an irradiance of 3 mW/cm(2) using a 370-nm UVA double diode. Groups of three animals were killed immediately after treatment and at 3 and 8 months, respectively. Biomechanical stress-strain measurements were performed using a microcomputer-controlled biomaterial tester on 4 x 10-mm corneal strips. Corneal thickness in the treated rabbit cornea was 408 +/- 20 microm. A constant and significant increase in ultimate stress (of 69.7-106.0%), Young's modulus of elasticity (of 78.4-87.4%) and a decrease in ultimate strain (of 0.57-78.4%) were found over a time period of up to 8 months after cross-linking treatment. Riboflavin/UVA-induced collagen cross-linking leads to a longterm increase in biomechanical rigidity which remains stable over time. These data support our previous longterm clinical observations and give hope that this new treatment will halt progressive keratoconus definitively.

Biomechanical and tissue handling property comparison of decellularized and cryopreserved tibialis anterior tendons following extreme incubation and rehydration

Little is known regarding the biomechanical profiles and tissue handling properties of decellularized and cryopreserved human tibialis anterior tendons prepared as allografts for ACL reconstruction. This study compared allografts prepared using two extremes of the same cryoprotectant incubation and rehydration technique with a standardly prepared control group. Porcine tibiae with similar apparent BMD were randomly divided into three groups of ten specimens. Paired tendons were randomly divided into two experimental groups: Group 1 = 8 h incubation/15 min rehydration; Group 2 = 2 h incubation/1 h rehydration. Group 3 (control) consisted of ten standardly prepared tendons with 20 min rehydration. Tissue handling properties were graded during allograft preparation using a modified visual analog scale. Similar diameter allografts were fixed in matched diameter extraction drilled tibial tunnels with 35 mm long, 1 mm > tunnel diameter bioabsorbable interference screws. Potted constructs were mounted in a servo hydraulic device, pretensioned between 10-50 N at 0.1 Hz (10 cycles), and isometric pretensioned at 50 N for 1 min, prior to 500 submaximal loading cycles (50-250 N) at 0.5 Hz, and load to failure testing (20 mm/min). Constructs prepared under extreme conditions generally displayed comparable biomechanical properties to the control condition. Group 1 (8 h incubation/15 min rehydration)(-34 +/- 35 ms) and Group 2 (2 h incubation/1 h rehydration) (-22 +/- 38 ms) displayed smaller mean displacement-load peak phase timing differences over the initial ten cycles compared to Group 3 (control)(-42 +/- 49 ms), P = 0.004, suggesting greater relative construct stiffness. Group 1 (8 h incubation/15 min rehydration) (234.9 +/- 34 N/mm) and Group 2 (2 h incubation/1 h rehydration)(231.3 +/- 43 N/mm) displayed lower construct stiffness during load to failure testing than Group 3 (control)(284.5 +/- 25.2 N/mm), P = 0.003. Group 1 (8 h incubation/15 min rehydration) differed from Group 2 (2 h incubation/1 h rehydration) and Group 3 (control) for perceived tensile stiffness (2.4 +/- 2.0 vs. 7.0 +/- 0.5 and 7.9 +/- 0.3, respectively), compressive resilience (1.7 +/- 0.8 vs. 5.9 +/- 1.0 and 7.8 +/- 0.4, respectively), handling ease (2.8 +/- 1.0 vs. 6.5 +/- 0.5 and 7.0 +/- 0.7, respectively), color (2.6 +/- 0.8 vs. 4.7 +/- 0.7 and 5.1 +/- 0.3, respectively) and texture (4.0 +/- 0.8 vs. 6.2 +/- 0.8 and 6.8 +/- 0.8, respectively) (P < 0.0001). Group 2 (2 h incubation/1 h rehydration)(6.0 +/- 0.7 and 5.9 +/- 1.0, respectively) also differed from Group 3 (control)(6.8 +/- 0.8 and 7.8 +/- 0.4, respectively) for general "feel" and compressive resilience (P < 0.0001). Tensile stiffness and compressive resilience displayed moderate and weak relationships, respectively with displacement during submaximal cyclic loading (r2 = 0.78 and 0.58, respectively), stiffness (r2 = 0.33 and 0.44, respectively) and load at failure (r2 = 0.59 and 0.37, respectively) for Group 3 (control), but not for experimental Group 1 (8 h incubation/15 min rehydration) or Group 2 (2 h incubation/1 h rehydration). Knee surgeons should be aware that soft tissue tendon decellularization and cryopreservation may change the biomechanical stiffness, tissue handling properties, and relationships between these variables compared to standardly prepared allograft tissue.

Results of the T2 humeral nailing system with special focus on compression interlocking

The outcome of interlocking nailing of humeral shaft fractures is controversial. Variations in implants, operative technique and follow-up parameters hinder comparative studies. The aim of this investigation was to evaluate the T2 humeral nailing system according to clinical results, and to recognise advantages and disadvantages of compression interlocking. A total of 36 consecutive humeral shaft fractures were treated with an interlocking humeral nail. In 23 cases the antegrade and in 14 cases the retrograde approach was used. Compression locking was performed in 21 and static locking in 15 cases. At 22-month follow-up, bone consolidation was present in 35 cases; in 5 cases complications developed. There was no significant difference between compression and static locking nor between the antegrade and retrograde approach regarding complications, Constant shoulder score, mean Morrey elbow score, SF-12 physical score or SF-12 mental score. Antegrade and retrograde interlocking nailing of humeral shaft fractures with the T2 nailing system can result in good functional outcome and unimpaired quality of life. Compression interlocking can minimise the fracture gap and increase the biomechanical stiffness. Potential disadvantages of compression interlocking include possible bending or loosening of the locking screw in the dynamic oblong hole. An additional static locking screw should be used on the humerus after compression interlocking.

Biophysikalische Grundlagen der Kollagenvernetzung

The reduced mechanical stability of the cornea in keratoconus or in keratectasia after Lasik may be increased by photooxidative cross-linking of corneal collagen. The biophysical principles are compiled for the safe and effective application of this new treatment method. The setting of the therapy parameters should be elucidated from the absorption behaviour of the cornea. The safety of the method for the endothelium cells and the lens will be discussed. The induced cross-links are shown to be the result of changes in the physico-chemical properties of the cornea. To reach a high absorption of the irradiation energy in the cornea, riboflavin of a concentration of 0.1% and UV light of a wavelength of 370 nm, corresponding to the relative maximum of absorption of riboflavin, were used. An irradiance of 3 mW/cm(2) and an irradiation time of 30 min lead to an increase of the mechanical stiffness. The endothelium cells will be protected due to the high absorption within the cornea, that means the damaging threshold of the endothelium cells will not be reached in a 400 microm thick stroma. As evidence for cross-links we can consider the increase of the biomechanical stiffness, the increased resistance against enzymatic degradation, a higher shrinkage temperature, a lower swelling rate and an increased diameter of collagen fibres. The therapy parameters were tested experimentally and have been proven clinically in the corneal collagen cross-linking. These parameters should be respected to reach a safe cross-linking effect without damage of the adjacent tissues.

Promotion of Medial Collateral Ligament Healing Using a High-Voltage Electrical Field

To assess the biomechanical and histological effects of a high-voltage electrical field on ligament healing. 54 male Wistar rats were used. The medial collateral ligaments (MCLs) of 6 rats were measured for normal biomechanical failure load and stiffness. MCLs of the remaining 48 rats were transected and randomly assigned; half were subjected to a high-voltage electrical field for 8 hours a day (experimental group), and the other half were kept under normal conditions (control group). Eight rats from each group were killed 2, 4, and 6 weeks after the transection and their MCLs were tested biomechanically and histologically. Failure load and stiffness of the MCLs in the experimental group were significantly higher than those in the control group 2 weeks after transection, but not thereafter. At week 2, the granulation tissue completely occupied the space between the cut ends of the MCL in the experimental group, whereas some gaps remained in the control group. A high-voltage electrical field appear to accelerate ligament healing in the initial 2 weeks. Earlier initial healing may allow earlier start of the rehabilitation.

Biomechanical Evaluation of Primary Stiffness of Tibiotalocalcaneal Fusion with Intramedullary Nails

Intramedullary implants are being used with increasing frequency for tibiotalocalcaneal fusion (TTCF). Clinically, the question arises whether intramedullary (IM) nails should have a compression mode to enhance biomechanical stiffness and fusion-site compression. This biomechanical study compared the primary stability of TTCF constructs using compressed and uncompressed retrograde IM nails and a screw technique in a bone model. For each technique, three composite bone models were used. The implants were a Biomet nail (static locking mode and compressed mode), a T2 femoral nail (compressed mode); a prototype IM nail 1 (PT1, compressed mode), a prototype IM nail 2 (PT2, dynamic locking mode and compressed mode), and a three-screw construct. The compressed contact surface of each construct was measured with pressure-sensitive film and expressed as percent of the available fusion-site area. Stiffness was tested in dorsiflexion and plantarflexion (D/P), varus and valgus (V/V), and internal rotation and external rotation (I/E) (20 load cycles per loading mode). Mean contact surfaces were 84.0 +/- 6.0% for the Biomet nail, 84.0 +/- 13.0% for the T2 nail, 70.0 +/- 7.2% for the PTI nail, and 83.5 +/- 5.5% for the compressed PT2 nail. The greatest primary stiffness in D/P was obtained with the compressed PT2, followed by the compressed Biomet nail. The dynamically locked PT2 produced the least primary stiffness. In V/V, PT1 had the (significantly) greatest primary stiffness, followed by the compressed PT2. The statically locked Biomet nail and the dynamically locked PT2 had the least primary stiffness in V/V. In I/E, the compressed PT2 had the greatest primary stiffness, followed by the PT1 and the T2 nails, which did not differ significantly from each other. The dynamically locked PT2 produced the least primary stiffness. The screw construct's contact surface and stiffness were intermediate. The IM nails with compression used for TTCF produced good contact surfaces and primary stiffness. They were significantly superior in these respects to the uncompressed nails and the screw construct. The large contact surfaces and great primary stiffness provided by the IM nails in a bone model may translate into improved union rates in patients who have TTCF.

A Biomechanical Study of 3 Different Types of Sublaminar Wire Used for Constructs in the Thoracic Spine

A biomechanical study was carried out on 3 different types of sublaminar wire used in constructs to secure the thoracic spine: stainless steel monofilament wire (steel wire), titanium cable (cable), and ultra-high molecular weight polyethylene tape (tape). Two experiments were carried out. Experiment 1: Thirty-one fresh human thoracic vertebrae classified as osteoporotic (bone mineral density of <0.8 g/cm2) were used. The steel wire, cable, or tape was placed sublaminarly and a tensile force was applied until the steel wire, cable, or tape cut 5 mm through the lamina, and the force at this point was noted. Experiment 2: Seven fresh human thoracic spines (T7-T10) were biomechanically tested as follows: axial compression (250 N), flexion (7.5 Nm), extension (7.5 Nm), left lateral bending (7.5 Nm), right lateral bending (7.5 Nm), left axial torsion (10 Nm), and right axial torsion (10 Nm). This sequence was applied to the intact spine. The spine was then de-stabilized and then restabilized using one or other of the 3 different types of sublaminar wires. The biomechanical testing was then repeated on the restabilized spine and stiffness curves were generated. In the laminar cut-through test, the cut-through force for tape was higher than that for either steel wire or cable. In the biomechanical stiffness testing, there was no significant difference between the 3 different sublaminar wiring constructs in any of the loading modes tested. The results of both experiments suggest that tape is as good, if not better, than steel wire or cable as a sublaminar wiring construct material.

Lumbar interbody fusion using autologous bone marrow mesenchymal stem cell-calcium phosphate ceramic composite in rhesus monkey

To determine the osteogenic capacity of autologous bone marrow mesenchymal stem cells (BMSCs)-calcium phosphate ceramic composites in vitro and implanted as a bone graft substitute for lumbar anterior interbody fusion in rhesus monkeys. From March 2003 to April 2005, 9 adult rhesus monkeys underwent lumbar L(3 - 4) and L(5 - 6) discectomy and interbody fusion via an anterior retroperitoneal approach. Two fusion sites in each animal were randomly assigned to two of three treatments: autogenous tricortical iliac crest bone graft (autograft group, n = 6) or cell-free ceramic graft (ceramic group, n = 6) or BMSCs-ceramic composite graft (BMSCs group, n = 6). Autologous BMSCs were culture-expanded and stimulated with osteogenic supplement. The cell-ceramic composites were constructed in a rotary dynamic cell culture system. The spinal fusion segments were evaluated by radiography, biomechanical testing, histologic analysis and histomorphometric analysis at 3 months post-surgery. Biomechanical testing showed that spinal segments from the autograft group and the BMSCs-ceramic group were statistically and significantly stiffer than the cell-free ceramic group. The BMSCs-ceramic group and the autograft group showed equivalent biomechanical stiffness by statistical analysis. Histologically, both the autograft group and the BMSCs-ceramic group achieved osseous union, but the cell-free ceramic group had a fibrous union. Quantitative histologic analysis showed that the amount of bone formation was significantly greater in the autograft group and the BMSCs-ceramic group compared with the cell-free ceramic group. However, the amount of ceramic residue was significantly greater in the cell-free ceramic group versus the BMSCs-ceramic group. The results indicate that BMSC-ceramic composites can enhance bone regeneration and achieve osseous spinal fusion 3 months after the implantation in rhesus monkey interbody fusion model. Cell-free ceramics has an unsatisfactory efficacy in spinal fusion due to its tense fibrous fusion.

A Biomechanical Assessment of Infra-Laminar Hooks as an Alternative to Supra-Laminar Hooks in Thoracolumbar Fixation

Biomechanical study, using human cadaveric spines, of two types of posterior fixation in the thoracolumbar spine. To compare fixation in the thoracolumbar spine using pedicle screws combined with infra-laminar hooks (at both the cranial and caudal ends of the same vertebra) with fixation using pedicle screws alone. The standard method of using laminar hooks is to place supra-laminar hooks at the level above the vertebra being fixed by the cranial pedicle screws, and infra-laminar hooks at the level of the vertebra receiving the caudal pedicle screws. However, using this method, a single normal motion segment is killed. In the proposed technique, infra-laminar hooks will be used at the same level as the cranial pedicle screws, thus sparing the superior adjacent segment. PEDICLE SCREW LOOSENING TEST: Nine vertebrae (T9-L2) had pedicle screws inserted into both pedicles. On one side, the pedicle screw was connected to a fixation rod, while on the other side the pedicle screw was combined with an infra-laminar hook and connected to a rod. The rod on one side was then pushed with increasing force until loosening of the screw. The same test was repeated on the other rod until loosening of the screw. The reverse test (i.e., pulling until loosening) was done in a similar manner on another nine vertebrae. BIOMECHANICAL STIFFNESS TEST: Eight spines (T12-L2) were biomechanically tested in: axial compression, flexion, extension, left and right lateral bending, and left and right axial torsion. This sequence was applied to: 1) the intact spine; 2) after applying pedicle screws and infra-laminar hooks with rods to the destabilized spine; and 3) after removal of the hooks (with the pedicle screw and rods still inserted). From the load-deformation curves obtained, stiffness values were calculated. PEDICLE SCREW LOOSENING TEST: The pedicle screw loosening tests showed that a pedicle screw combined with an infra-laminar hook offers significantly greater fixation strength as compared to a pedicle screw alone. Biomechanical stiffness test. Fixation by pedicle screws combined with infra-laminar hooks, placed on the vertebra fixed by pedicle screws at both the cranial and caudal ends, offered a stiffer construct as compared to fixation by pedicle screws alone. This method offers a firm construct without sacrificing a normal motion segment.

Augmentation of Anterior Lumbar Interbody Fusion with Anterior Pedicle Screw Fixation: Demonstration of Novel Constructs and Evaluation of Biomechanical Stability in Cadaveric Specimens

Anterior lumbar interbody fusion (ALIF) has proven effective for indications including discogenic back pain, nonunion, and instability. Current practice involves posterior pedicle screw augmentation of the ALIF procedure (ALIF-PPS). This approach requires intraoperative repositioning of the patient for percutaneous posterior pedicle screw placement. We have developed a novel technique in which the ALIF procedure is augmented with anterior pedicle screws (APS; ALIF-APS). In this study, we introduce this new technique and compare the biomechanical stability of the novel ALIF-APS with the current standard ALIF-PPS. The technique was demonstrated in a cadaveric L4-S1 specimen using neuronavigation and fluoroscopy. Plain radiographs and computed tomographic scans of the construct were obtained. Twelve cadaveric spines (7 men and 5 women) from donors with an average age of 81 years (range, 64-93 yr) were then harvested from L4-S1. Six specimens were dedicated to ALIF-APS constructs, and the remaining six were dedicated to ALIF-PPS constructs. The specimens were then studied at L5-S1 in the following steps: 1) intact form, 2) after anterior discectomy, 3) after implantation of titanium cages (ALIF), and 4) after APS or PPS fixation in conjunction with the ALIF. Measurements were obtained in axial rotation and left and right lateral bending flexion-extension. Data were normalized by calculating the ratio of the stiffness of the instrumented to the intact spine. Statistical analyses were then performed on the data. Radiographs and computed tomographic scans of the construct showed accurate placement of the APS at L5 and S1. The normalized data showed that ALIF-APS and ALIF-PPS had approximately equal stability in axial rotation (1.17 +/- 0.43 versus 0.85 +/- 0.14), lateral bending (0.93 +/- 0.22 versus 0.95 +/- 0.16), and flexion- extension (0.77 +/- 0.13 versus 0.84 +/- 0.2). Paired t test analysis did not show a significant difference between the biomechanical stiffness of ALIF-APS and ALIF-PPS in axial rotation, lateral bending, and flexion-extension. We demonstrate a new technique in a cadaveric specimen whereby the ALIF procedure is augmented with APS fixation using neuronavigation and fluoroscopy. Biomechanical evaluation of the constructs suggests that the ALIF-APS has comparable stability with ALIF-PPS. APS augmentation of ALIF has potential advantages over the current standard ALIF-PPS because it can 1) eliminate the patient repositioning step, 2) minimize the total number of incisions and the total operative time, and 3) protect against dislocation of the ALIF interbody graft or cage. Work is in progress to develop a low-profile system for the novel APS constructs described here.

Polyetheretherketone as a biomaterial for spinal applications

Threaded lumbar interbody spinal fusion devices (TIBFD) made from titanium have been reported to be 90% effective for single-level lumbar interbody fusion, although radiographic determination of fusion has been intensely debated in the literature. Using blinded radiographic, biomechanic, histologic, and statistical measures, we evaluated a radiolucent polyetheretherketone (PEEK)-threaded interbody fusion device packed with autograft or rhBMP-2 on an absorbable collagen sponge in 13 sheep at 6 months. Radiographic fusion, increased spinal level biomechanical stiffness, and histologic fusion were demonstrated for the PEEK cages filled with autograft or rhBMP-2 on a collagen sponge. No device degradation or wear debris was observed. Only mild chronic inflammation consisting of a few macrophages was observed in peri-implant tissues. Based on these results, the polymeric biomaterial PEEK may be a useful biomaterial for interbody fusion cages due to the polymer's increased radiolucency and decreased stiffness.

Evaluation of Autologous Bone Marrow Mesenchymal Stem Cell–Calcium Phosphate Ceramic Composite for Lumbar Fusion in Rhesus Monkey Interbody Fusion Model

Autologous bone marrow mesenchymal stem cell (BMSC)-calcium phosphate ceramic composites were constructed in vitro and implanted as a bone graft substitute for lumbar anterior interbody fusion in rhesus monkeys to determine the osteogenic capacity of the composites. Nine adult rhesus monkeys underwent lumbar L3-L4 and L5-L6 diskectomy and interbody fusion via an anterior retroperitoneal approach. Two fusion sites in each animal were randomly assigned to two of three treatments: autogenous tricortical iliac crest bone graft (autograft group), cell-free ceramic graft (ceramic group), or BMSC-ceramic composite graft (BMSC group). Autologous BMSCs were expanded in culture and stimulated with osteogenic supplement. The spinal fusion segments were evaluated by radiography, biomechanical testing, histologic analysis, and histomorphometric analysis 3 months postsurgery. The BMSC group achieved lumbar interbody fusion superior to that of the ceramic group, both biomechanically and histologically. The BMSC group and the autograft group showed equivalent biomechanical stiffness. Ceramic residues were significantly greater in the ceramic group versus the BMSC group. The results indicate that BMSC-ceramic composites can enhance bone regeneration and achieve osseous spinal fusion 3 months after implantation in the rhesus monkey interbody fusion model.

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 4. Morphometric, microscopic and biomechanical properties of the digital tendons of the forelimb

AIM: To determine the weight, volume, density and cross-sectional area (CSA) of the digital flexor tendons, common digital extensor tendon (CDET) and suspensory ligament (SL) of the forelimb of young Thoroughbred horses in early training, and to assess the response to a training programme of known parameters of superficial digital flexor tendon (SDFT) tissue at mid-metacarpal level. METHODS: The tendons of seven 2-year-old Thoroughbred horses in training were inspected, transected into segments of known length, and compared with those from seven untrained horses matched for age, sex and breed. The weight, volume, density and CSA of each segment, and the crimp angle, histological features, and biomechanical ultimate stress and stiffness of tendon samples from the mid-metacarpal region of the SDFT were determined. RESULTS: There was no macroscopic evidence of swelling or discolouration in any of the tendon segments or cut surfaces. The volumes of SDFT and CDET segments of horses in the trained group were significantly greater than those in the untrained group (p=0.036 and p=0.039, respectively). A greater increase in volume than weight resulted in a lower density in the SDFT but not CDET in trained compared with untrained horses (p=0.038). CSA of these two tendons was significantly greater in the trained group (p=0.002 and 0.036, respectively), the percentage increase being greater in the CDET than the SDFT. The number of tenocytes at four sites in the mid-metacarpal SDFT region was less in trained than untrained horses (p=0.025). There was no histological evidence of inflammation, and no difference in crimp angle between groups. There was no significant between-group difference in stiffness or ultimate stress of tendon strips. CONCLUSIONS: Volume and CSA of the SDFT and CDET were larger in trained than untrained horses. The SDFT was less dense in the trained group. Because no evidence of tendonitis was detected and training appeared to have no significant effect on crimp angle or biomechanical properties of tendon strips, the size and density changes were presumed to be adaptive and induced by the training. CLINICAL RELEVANCE: Although evident in this in vitro study, the detection of adaptive from initial pathological increase in size of the SDFT is likely to be difficult in vivo.

Bioabsorbable Interbody Cages in a Sheep Cervical Spine Fusion Model

An experimental study using a sheep cervical spine interbody fusion model. To compare interbody fusion of an autologous tricortical iliac crest bone graft with two bioabsorbable cages and to determine whether there are differences between the three interbody fusion techniques in 1) the ability to preserve postoperative distraction, 2) the biomechanical stability, and 3) the histologic characteristics of intervertebral bone matrix formation. Bioabsorbable cages would be beneficial compared with metallic cages; however, currently no suitable bioabsorbable interbody fusion cage is available. Twenty-four sheep underwent C3/C4 discectomy and fusion. The following stabilization techniques were used: Group 1) autologous tricortical iliac crest bone graft (n = 8); Group 2) bioabsorbable cage made of 70/30 poly(l-lactide-co-d,l-lactide) (experimental) filled with autologous cancellous bone graft (n = 8); Group 3) bioabsorbable cage made of a polymer-calciumphosphate composite (Biomet Europe, Dordrecht, The Netherlands) filled with autologous cancellous bone graft (n = 8). Radiographic scans to determine disc space height were performed before and after surgery and after 1, 2, 4, 8, and 12 weeks, respectively. After 12 weeks, animals were killed, and fusion sites were evaluated using functional radiographic views in flexion and extension. Quantitative computed tomographic scans were used to assess bone mineral density, bone mineral content, and bony callus volume. Biomechanical testing was performed in flexion, extension, axial rotation, and lateral bending to determine stiffness, ROM, and neutral and elastic zone. Histomorphological and histomorphometrical analysis were performed to evaluate fusion and foreign body reactions associated with the bioabsorbable cages. Over a 12-week period, the polymer-calciumphosphate composite cage showed significantly higher values for disc space height compared with the bone graft and the poly(l-lactide-co-d,l-lactide) cage. Additionally, the polymer-calciumphosphate composite cage demonstrated a significantly higher stiffness and lower ROM, neutral zone, and elastic zone in axial rotation and lateral bending than any other group. However, quantitative computed tomographic scans demonstrated cracks in six of the eight polymer-calciumphosphate composite cages after 12 weeks. Histologically, the highest bone volume/total volume ratio and the highest fusion rate were found in the polymer-calciumphosphate composite cage group. Although the poly(l-lactide-co-d,l-lactide) cage showed grade I through III foreign body reactions in all fusion areas, only two animals developed grade I foreign body reactions with the polymer-calciumphosphate composite cage. After 12 weeks, there was no significant difference between the bioabsorbable poly(l-lactide-co-d,l-lactide) cage and the tricortical bone graft. In comparison to the tricortical bone graft, the bioabsorbable polymer-calciumphosphate composite cage showed significantly better distractive properties, a significantly higher biomechanical stiffness, and an advanced interbody fusion; however, six of eight polymer-calciumphosphate composite cages cracked. Although the fate of the foreign body reactions and the cracks is currently unclear for both bioabsorbable cages, the early appearance of large osteolysis associated with use of the poly(l-lactide-co-d,l-lactide) cage allows skepticism regarding the value of this bioabsorbable implant.