Allogeneic mesenchymal precursor cells (MPCs) combined with an osteoconductive scaffold to promote lumbar interbody spine fusion in an ovine model.

BACKGROUND: Advances in immunomagnetic cell sorting have enabled isolation and purification of pleuripotent stem cells from marrow aspirates and have expanded stem cell therapies to include allogeneic sources. PURPOSE: This study aimed to determine the safety and efficacy of allogeneic mesenchymal precursor cells (MPCs) combined with an osteoconductive scaffold in lumbar interbody spinal fusion using an ovine model. STUDY DESIGN: Thirty-two skeletally mature ewes underwent a single-level interbody fusion procedure using a Polyetheretherketone fusion cage supplemented with either iliac crest autograft (AG) or an osteconductive scaffold (Mastergraft Matrix, Medtronic, Memphis, TN, USA) with 2.5×10(6) MPCs, 6.25×10(6) MPCs, or 12.5×10(6) MPCs. METHODS: Plain radiographs and computed tomography scans were scored for bridging bone at multiple points during healing and at necropsy. The biomechanical competency of fusion was scored by manual palpation and quantified using functional radiographs at necropsy. Postnecropsy histopathology and histomorphometric analysis assessed the local response to MPC treatment and quantified the volume and connectivity of newly formed bridging bone. Safety was assessed by serum biochemistry, hematology, and organ histopathology. RESULTS: Mesenchymal precursor cell treatment caused no adverse systemic or local tissue responses. All analyses indicated MPCs combined with an osteoconductive scaffold achieved similar or better fusion success as AG treatment after 16 weeks, and increasing the MPC dose did not enhance fusion. Manual palpation of the fusion site indicated more than 75% of MPC-treated and 65% of AG-treated animals achieved rigid fusion, which was corroborated with functional radiography. Computed tomography fusion scores indicated all animals in the MPC- and AG-treatment groups were fused at 16 weeks, yet X-ray scores indicated only 67% of the AG-treated animals were fused. Histomorphometry analyses showed equivalent outcomes for fusion connectivity and bony fusion area for MPC- and AG-treated groups. Approximately 6% residual graft material remained in the MPC-treated fusion sites at 16 weeks. CONCLUSIONS: Adult allogeneic MPCs delivered using an osteoconductive scaffold were both safe and efficacious in this ovine spine interbody fusion model. These results support the use ofallogeneic MPCs as an alternative to AG for lumbar interbody spinal fusion procedures.

Allogeneic mesenchymal progenitor cells for posterolateral lumbar spine fusion in sheep.

BACKGROUND CONTEXT: Osteoconductive porous ceramic bone graft materials supplemented with mesenchymal precursor cells (MPC) derived from autologous bone marrow aspirates have been shown to stimulate successful interbody and posterolateral spine fusion in preclinical models. Recent advances in immunomagnetic cell sorting have enabled purification and isolation of pluripotent stem cells from marrow aspirates and have expanded stem cell technology to allogeneic cell sources. Allogeneic MPC technology combined with appropriate synthetic biomaterial carriers could provide both the osteogenic and osteoconductive components needed for successful posterolateral spine fusion without the need for autologous bone harvest or expensive recombinant protein technology. PURPOSE: To determine the safety and efficacy of a hydroxyapatite:tricalcium phosphate graft material supplemented with allogeneic mesenchymal precursor cells in posterolateral lumbar spine fusion using an ovine model. STUDY DESIGN: Skeletally mature ewes underwent single-level instrumented posterolateral lumbar spine fusion using either autograft (AG), hydroxyapatite:tricalcium phosphate carrier (CP), or CP supplemented with allogeneic mesenchymal progenitor cells (MPCs). Three doses of MPCs were evaluated: 25 × 106 cells (low dose, LD), 75 × 106 cells (mid dose, MD), and 225 × 106 cell (high dose, HD). Animals survived for either 4 or 9 months. METHODS: Plain radiographs were acquired and scored for bridging bone at regular intervals during healing to monitor fusion development. Hematology, coagulation, and serum chemistry were monitored at regular intervals throughout the study to monitor animal health. After necropsy, computed tomography, high-resolution radiography, biomechanical testing, organ pathology, bone histopathology, and bone histomorphometry were conducted to monitor the safety and ascertain the efficacy of MPC treatment. RESULTS: MPC treatment in this spine fusion model resulted in no observed adverse systemic or local tissue responses. Radiographically, fusion scores for MPC-treated animals were uniformly higher compared with those treated with carrier alone (CP) after 3 months and continued the same trend throughout 9 month of healing. Quantitative computed tomography confirmed better connectivity of the fusion for MPC treatment groups compared with CP. Biomechanical analyses were not able to differentiate between treatment groups. Histomorphometry results confirmed radiographic and quantitative computed tomography results; cell-supplemented treatment groups and autograft had equivalent amounts of bone within the fusion mass and less bony fusion tissue was found within the fusion mass in specimens from the CP treatment group. No conclusive effects of cell dose of fusion efficacy were noted. CONCLUSIONS: Adult allogeneic mesenchymal precursor cells delivered via a hydroxyapatite:tricalcium phosphate carrier were both safe and efficacious in this ovine spine fusion model. Results from this preclinical study support that allogeneic mesenchymal precursor cells produced fusion efficacy similar to that achieved using iliac crest autograft, thereby providing a safe and viable option to achieve successful posterolateral spine fusion.

Regeneration of periodontal tissues using allogeneic periodontal ligament stem cells in an ovine model.

AIM: To investigate the capacity of allogeneic periodontal ligament stem cells (PDLSCs) to regenerate periodontal tissues using an ovine periodontal defect model. MATERIALS & METHODS: Surgically created zero-wall dehiscence periodontal defects created in Merino sheep were filled with 1 × 10(7) allogeneic PDLSCs attached to Gelfoam(®), Gelfoam alone or left untreated. After 4 weeks, histological analysis was performed to assess periodontal regeneration. RESULTS: Allogeneic PDLSCs were well tolerated by recipient animals. The mean area of new alveolar bone was significantly greater in the PDLSC + Gelfoam treatment group compared with the defect-alone group. The PDLSC + Gelfoam and Gelfoam-only treatment groups displayed significantly greater length of new cementum and percentage of cementum regrowth compared with the defect-alone group. New Sharpey's fibers were generally more organized and significantly thicker within the PDLSC + Gelfoam treatment group. The PDLSC + Gelfoam treatment group also showed a trend of increased Sharpey's fiber attachment length compared with the Gelfoam-only and defect-alone groups. CONCLUSION: These studies support the potential use of allogeneic PDLSC preparations as viable therapies for periodontal regeneration in the clinical setting.

Effects of (60)Co gamma radiation dose on initial structural biomechanical properties of ovine bone--patellar tendon--bone allografts.

Gamma radiation is established as a procedure for inactivating bacteria, fungal spores and viruses. Sterilization of soft tissue allografts with high dose (60)Co gamma radiation has been shown to have adverse effects on allograft biomechanical properties. In the current study, bone-patellar tendon-bone (BPTB) allografts from 32 mature sheep were divided into two treatment groups: low-dose radiation at 15 kGy (n = 16) and high-dose radiation at 25 kGy (n = 16) with the contralateral limb serving as a 0 kGy (n = 32) non-irradiated control. Half of the tendons from all treatment groups were biomechanically tested to determine bulk BPTB mechanical properties, cancellous bone compressive properties, and interference screw pull-out strength. The remaining tissues were prepared, implanted, and mechanically tested in an acute in vitro anterior crucial ligament (ACL) reconstruction. Low-dose radiation did not adversely affect mechanical properties of the tendon allograft, bone, or ACL reconstruction compared to internal non-irradiated control. However, high-dose radiation compromised bulk tendon load at failure and ultimate strength by 26.9 and 28.9%, respectively (P < 0.05), but demonstrated no negative effect on the cancellous bone compressive properties or interference screw pull-out strength. Our findings suggest that low dose radiation (15 kGy) does not compromise the mechanical integrity of the allograft tissue, yet high dose radiation (25 kGy) significantly alters the biomechanical integrity of the soft tissue constituent.

Gene therapy to enhance allograft incorporation after host tissue irradiation.

Structural bone allografts are used to reconstruct large skeletal defects after tumor surgery. Although allograft-related complications are declining, the use of perioperative radiation therapy is associated with a poorer outcome. Recently, BMP-2 levels in the host bed were reportedly diminished after exposure to radiation doses consistent with those used perioperatively to treat musculoskeletal sarcoma. Reintroduction of this osteogenic protein may circumvent the deleterious effects of preoperative radiation on allograft incorporation. We introduced a novel polymeric BMP-2 gene delivery system into the host-allograft junctions at the time of transplantation in an ovine tibial defect model with or without preoperative exposure to 50 Gy radiation. After 4 months, we noted no radiographic or histologic improvements in allograft incorporation after preoperative radiation and BMP-2 reintroduction; however, 50 Gy radiation was associated with increased porosity in the interface regions and poorer radiographic healing. We identified no BMP2-expressing cells or protein in the interface at the study end point, suggesting the polymeric gene delivery system was unable to promote extended expression of the protein or induce a healing response. Although gene therapy may hold promise as a novel technique to improve allograft incorporation, our data do not support that contention with the current approach.

Effects of low intensity pulsed ultrasound with and without increased cortical porosity on structural bone allograft incorporation.

BACKGROUND: Though used for over a century, structural bone allografts suffer from a high rate of mechanical failure due to limited graft revitalization even after extended periods in vivo. Novel strategies that aim to improve graft incorporation are lacking but necessary to improve the long-term clinical outcome of patients receiving bone allografts. The current study evaluated the effect of low-intensity pulsed ultrasound (LIPUS), a potent exogenous biophysical stimulus used clinically to accelerate the course of fresh fracture healing, and longitudinal allograft perforations (LAP) as non-invasive therapies to improve revitalization of intercalary allografts in a sheep model. METHODS: Fifteen skeletally-mature ewes were assigned to five experimental groups based on allograft type and treatment: +CTL, -CTL, LIPUS, LAP, LIPUS+LAP. The +CTL animals (n = 3) received a tibial ostectomy with immediate replacement of the resected autologous graft. The -CTL group (n = 3) received fresh frozen ovine tibial allografts. The +CTL and -CTL groups did not receive LAP or LIPUS treatments. The LIPUS treatment group (n = 3), following grafting with fresh frozen ovine tibial allografts, received ultrasound stimulation for 20 minutes/day, 5 days/week, for the duration of the healing period. The LAP treatment group (n = 3) received fresh frozen ovine allografts with 500 mum longitudinal perforations that extended 10 mm into the graft. The LIPUS+LAP treatment group (n = 3) received both LIPUS and LAP interventions. All animals were humanely euthanized four months following graft transplantation for biomechanical and histological analysis. RESULTS: After four months of healing, daily LIPUS stimulation of the host-allograft junctions, alone or in combination with LAP, resulted in 30% increases in reconstruction stiffness, paralleled by significant increases (p < 0.001) in callus maturity and periosteal bridging across the host/allograft interfaces. Longitudinal perforations extending 10 mm into the proximal and distal endplates filled to varying degrees with new appositional bone and significantly accelerated revitalization of the allografts compared to controls. CONCLUSION: The current study has demonstrated in a large animal model the potential of both LIPUS and LAP therapy to improve the degree of allograft incorporation. LAP may provide an option for increasing porosity, and thus potential in vivo osseous apposition and revitalization, without adversely affecting the structural integrity of the graft.

A mechanical and computational investigation on the effects of conduit orientation on the strength of massive bone allografts.

Structural bone allografts are used to reconstruct large skeletal defects resulting from trauma, tumor resection, or revision arthroplasty. Though used for over a century, bone allografts suffer from a high rate of mechanical failure due to limited graft revitalization even after extended periods in vivo. The current study evaluated the mechanical properties of longitudinally perforated cortical bone allografts (LAP) that have been shown to promote accelerated graft incorporation in a large animal model. The compressive and tensile properties of longitudinally perforated allograft specimens, as determined through uniaxial compression and diametral compression tests, respectively, were not significantly affected by the presence of the conduit. However, transversely perforated grafts (TAP) demonstrated a marked decrease in tensile capacity (p=0.04). Finite element analysis demonstrated moderate increases in the maximum principal stresses in LAP specimens while TAP models indicated an 83.4% increase in maximum principle stress near the conduit on the endosteal surface of the graft. This research and the previous in vivo study suggest that LAP adequately serves as an internal template within the cortical bone allograft for osseous apposition and revitalization without adversely affecting the structural or mechanical integrity of the graft.

Hollow calcium phosphate microcarriers for bone regeneration: in vitro osteoproduction and ex vivo mechanical assessment.

Synthetic grafting materials, such as calcium phosphates (hydroxyapatite, HA; tricalcium phosphate, TCP), polymers, or composites thereof, can be used as osteoconductive scaffolds and delivery vehicles for osteoinductive growth factors. Carrier materials must be engineered to deliver these factors in a controlled fashion at a rate and dose consistent with the biological need and responsiveness of the system to optimize bone formation and ingrowth. They should also simultaneously provide mechanical support and slowly resorb as new bone is formed. This investigation assessed the elution characteristics of BMP-7 (OP-1) from hollow calcium phosphate spheres of varying chemical composition (HA/beta-TCP) and porosity (dense/porous). The pharmacokinetics indicated a bimodal trend of protein release with protein elution peaking between fifteen and thirty minutes in solution (bolus release) and continuing through the eight-week time point (sustained release). Eluted OP-1 bioactivity was characterized over a three-week period using mesenchymal stem cell (MSC) cultures and included assessment of the protein's differential, proliferative, and calcified nodule forming abilities. Alkaline phosphatase enzyme (ALP) activity in MSCs peaked between 12 and 16 days post-OP-1 exposure. Elutant from the HA dense treatment group induced the highest degree of ALP expression while elutant from the beta-TCP treatment groups induced the formation of significantly higher numbers of calcified nodules in culture. The aggregate modulus of a clinically relevant 2 cc dose of carriers was quantified using custom designed testing fixtures to investigate the effects of carrier size, porosity, chemical composition, and the presence of a central hole on mechanical integrity. Significant increases in moduli were noted for carrier size and chemical composition (HA>beta-TCP). These preliminary in vitro and ex vivo results indicate the clinical potential of the hollow calcium phosphate carriers as successful load-bearing delivery vehicles for OP-1.

Efficacy of silicated calcium phosphate graft in posterolateral lumbar fusion in sheep.

BACKGROUND CONTEXT: Conditions requiring posterior lumbar spinal fusion remain a clinical challenge. Achieving arthrodesis using autogenous bone graft is inconsistent when rigid internal fixation such as transpedicular instrumentation is applied. Synthetic materials, particularly calcium phosphate-based ceramics, have shown promise for spine fusion applications, especially when combined with autograft. Silicate substitution has been shown to enhance the bioactivity of calcium phosphates and may obviate the need for autologous supplementation. PURPOSE: Determine efficacy of silicated calcium phosphate (Si-CaP) compared with autograft to generate solid lumbar fusion. STUDY DESIGN: Comparison of healing of instrumented posterolateral lumbar fusion in ewes at 2 and 6 months using Si-CaP or iliac crest autograft. METHODS: Eighteen skeletally mature ewes underwent implantation of either autograft or Si-CaP in the space spanning the L4-L5 transverse process. In vivo quantitative computed tomography (CT) scans were made at 2-month intervals and after euthanasia. Harvested spine segments were radiographed and biomechanically tested in bending at 6 months. Histological assessments were made at 2 and 6 months. RESULTS: Animals receiving Si-CaP graft were biomechanically and radiographically equivalent to those receiving autograft. Fusion mass density and volume were higher for the Si-CaP group throughout the healing period. Si-CaP regenerated normal bone tissue morphology, cellularity, and maturation with no inflammatory responses despite the fact that no autograft, bone marrow aspirate, or blood was mixed with the material. Histomorphometrically, fusion mass was higher for Si-CaP and bony bridging was equivalent when compared with autograft treatment. CONCLUSIONS: Si-CaP was biomechanically, radiographically, and histologically equivalent to autograft in generating a solid, bony, intertransverse process fusion in an ovine model. Both treatment groups achieved 100% bridging fusion after 6 months of healing.

In vivo BMP-7 (OP-1) enhancement of osteoporotic vertebral bodies in an ovine model.

BACKGROUND CONTEXT: Prevention of osteoporotic vertebral fractures could help at-risk individuals avoid the pain and morbidity associated with these fractures. Currently, patients with osteoporosis are treated with systemic medications to reduce fracture risk. Although effective, these therapies do not eliminate fractures and also tend to have a gradual time-dependent effect on fracture risk. The mechanism of action of the bone morphogenetic protein (BMP) family theoretically makes these molecules candidates for rapidly enhancing local bone structure. STUDY DESIGN: An in vivo study analyzing the effects of BMP-7 (osteogenic protein 1 [OP-1]) treatment on osteopenic ovine vertebral architecture and biomechanics. PURPOSE: We tested the hypothesis that local injection of OP-1 into osteopenic ovine vertebrae will improve bone mass and trabecular distribution, thereby reducing bone fragility and fracture risk. We specifically evaluated compressive biomechanics and morphology of osteopenic ovine vertebral bodies 6 months after local OP-1 treatment. STUDY DESIGN: In vivo animal study. METHODS: Skeletally mature sheep (n=24) underwent ovariectomy and were placed on low cation relative to anion diet. These interventions reduce bone density and induce skeletal fragility. After 6 months, sheep were randomly assigned to six treatment groups based on OP-1 dose (370 mg or 0 mg) and carrier with 4 animals/treatment group. Carriers A and B were poly-L-glycolic acid (PLGA) biospheres with different release kinetics (B allowing sustained BMP release); Carrier C was carboxymethylcellulose. After creating an 8-mm-diameter defect in the midvertebral body, sheep underwent intravertebral body implantation at two nonadjacent levels. Animals were euthanized 6 months after implantation and bone mineral density (BMD), biomechanics, and histomorphometry were assessed. Two-way analysis of variance was used to determine effects of OP-1 (alpha=0.05). RESULTS: An 81.9%, 333.2%, and 39.9% increase in stiffness was seen for OP-1 treated vertebra with Carriers A, B, and C respectively. Although these effects did not reach statistical significance, trends toward improvement were evident. Histology showed varied degrees of bony healing in the injection sites. Histomorphometrically, OP-1 treated vertebrae showed improvements in percent bone of up to 38% and star volume of up to 55% (with Carrier B). Improvements in whole vertebral body BMD were not detected for any treatment. CONCLUSION: In this study, local OP-1 treatment showed a positive trend in improving mechanical strength and histomorphometric parameters of osteopenic vertebra, despite the absence of consistent change in BMD. Controlled slow release of OP-1 using PLGA microspheres appeared to be the most effective method of protein delivery. In conclusion, we feel that the pilot data suggest that the use of OP-1 in the treatment of vertebral osteoporosis in an attempt to enhance bone strength merits further study.

Cortical bone graft and endoprosthesis in the distal radius of dogs: a biomechanical comparison of two different limb-sparing techniques.

OBJECTIVE: To compare the biomechanical properties of cortical bone and surgical steel endoprosthesis for limb-sparing surgery of the distal radius in dogs and evaluate the role of the ulna in providing stability to the reconstructed limb. STUDY DESIGN: Cadaveric biomechanical study. ANIMALS: Twelve pairs of normal canine thoracic limbs. METHODS: Paired limbs were divided into 4 groups: endoprosthesis and cortical bone graft, with and without preservation of the ulna. In each limb pair, the distal segment of the radius resected from the limb to be reconstructed with an endoprosthesis was used as the cortical bone graft in the contralateral limb. The ulna was resected en bloc with the radius and at the same level as the radial osteotomy in limbs where the ulna was not preserved. Limbs were tested in axial loading until failure. The load-deformation curve was used to acquire the biomechanical properties of each construct, which were compared using 2-way ANOVA. Failure modes were compared descriptively. RESULTS: Limbs reconstructed with the endoprosthesis had significantly greater yield load, energy at yield, and ultimate load compared with limbs reconstructed with a cortical bone graft. There were no significant differences in either energy to failure or stiffness between the 2 constructs. Preservation of the ulna did not significantly improve any of the biomechanical properties tested with either endoprosthesis or cortical bone graft constructs. The modes of failure in all 4 groups were variable and inconsistent. CONCLUSIONS: Limbs reconstructed with an endoprosthesis were biomechanically superior to limbs reconstructed with a cortical bone graft in axial loading to failure. Preservation of the ulna is not required to improve the stability in axial compression after limb-sparing surgery of the distal radius. CLINICAL RELEVANCE: The endoprosthesis may provide another option for limb-sparing surgery of the distal radius in dogs. It has potential advantages when compared with cortical bone grafts, including better biomechanical performance and resistance to implant failure in axial compression, immediate availability, and no requirement for bone banking facilities. The ulna can be resected en bloc with the radius without having a negative impact on construct stability. En bloc resection of the ulna and radius may decrease the risk of local tumor recurrence after limb-sparing surgery.

Ex vivo comparison of one versus two distal screws in 8 mm model 11 interlocking nails used to stabilize canine distal femoral fractures.

OBJECTIVE: To compare the structural properties of an 8 mm model 11 interlocking nail (IN) with 2 proximal and 2 distal screws (2/2) to 2 proximal and 1 distal screws (2/1) in an unstable canine fracture model. STUDY DESIGN: Ex vivo biomechanical investigation. SAMPLE POPULATION: Eight pairs of adult canine femurs. METHODS: A simple transverse distal metaphyseal femoral fracture with a 1 cm gap was created. The unstable fracture in 1 femur was repaired with a nail with 2 distal and 2 proximal screws and the paired femur with a nail with 1 distal and 2 proximal screws. Cyclic mechanical testing in torsion was performed to assess fatigue life, peak torque, stiffness, and mode of failure. RESULTS: All 2/1 IN-femoral constructs, but only 2 of eight 2/2 constructs, failed before completion of 50,000 loading cycles. The 2/2 constructs had significantly greater peak torque to failure (P = .002) and longer fatigue life (P = .00003) compared with 2/1 constructs. There were no significant differences in stiffness between 2/2 and 2/1 constructs when the non-failed constructs were compared (P > .5). All constructs failed by screw deformation. CONCLUSIONS: An 8 mm model 11 IN used for fixation of unstable canine distal femoral fractures has a longer fatigue life and is stronger under torsional loads when 2 rather than 1 distal screws are placed. CLINICAL RELEVANCE: When repairing unstable canine distal femoral fractures with an IN system, 2 distal screws should be inserted to avoid catastrophic implant failure before bone healing is achieved.

Evaluation of subchondral bone mineral density associated with articular cartilage structure and integrity in healthy equine joints with different functional demands.

OBJECTIVE: To determine and correlate subchondral bone mineral density and overlying cartilage structure and tensile integrity in mature healthy equine stifle (low magnitude loading) and metacarpophalangeal (high magnitude loading) joints. ANIMALS: 8 healthy horses, 2 to 3 years of age. PROCEDURE: Osteochondral samples were acquired from the medial femoral condyle (FC) and medial trochlear ridge (TR) of the stifle joint and from the dorsal (MC3D) and palmar (MC3P) aspects of the distal medial third metacarpal condyles of the metacarpophalangeal joint. Articular cartilage surface fibrillation (evaluated via India ink staining) and tensile biomechanical properties were determined. The volumetric bone mineral density (vBMD) of the underlying subchondral plate was assessed via dual-energy x-ray absorptiometry. RESULTS: Cartilage staining (fibrillation), tensile moduli, tensile strength, and vBMD were greater in the MC3D and MC3P locations, compared with the FC and TR locations, whereas tensile strain at failure was less in MC3D and MC3P locations than FC and TR locations. Cartilage tensile moduli correlated positively with vBMD, whereas cartilage staining and tensile strain at failure correlated negatively with vBMD. CONCLUSIONS AND CLINICAL RELEVANCE: In areas of high joint loading, the subchondral bone had high vBMD and the articular cartilage surface layer had high tensile stiffness but signs of structural wear (fibrillation and low failure strain). The site-dependent variations and relationships in this study support the concept that articular cartilage and subchondral bone normally adapt to physiologic loading in a coordinated way.

Radial nerve excursion and strain at the elbow and wrist associated with upper-extremity motion.

PURPOSE: This study evaluated the excursion necessary to accommodate common motions of daily living and associated strain on the radial nerve. The radial nerve was evaluated at the wrist and proximal to the elbow before it bifurcated. METHODS: Five fresh-frozen transthoracic cadaver specimens (10 arms) were dissected; the radial nerve was exposed at the elbow and wrist only enough to be marked with a microsuture. Excursion was measured using a laser mounted on a caliper fixed to the bone and aligned in the direction of nerve motion. Strain was measured with a device applied to the nerve at the elbow. Nerve excursion associated with motion of the shoulder, elbow, wrist, and fingers (measured by a goniometer) was assessed at the wrist and elbow. RESULTS: An average of 4.3 mm of radial nerve excursion was required at the wrist to accommodate wrist motion from 15 degrees of radial deviation to 30 degrees of ulnar deviation and 8.8 mm was needed for elbow motion from 10 degrees to 90 degrees . The radial nerve at the elbow experienced a 28% strain associated with the same motion of flexion and extension at the elbow. When all the motions of the wrist, fingers, elbow, and shoulder were combined 9.4 mm of radial nerve excursion was required at the wrist and 14.2 mm at the elbow. CONCLUSIONS: Any factor that limits excursion at these sites could result in repetitive traction of the nerve and possibly could play a role in the pathophysiology of a mechanical neuropathy, which in the case of the radial nerve most often manifests as pain.

Allograft bone decreases in strength in vivo over time.

Allograft bone is the primary source of graft material for structural oncological limb salvage procedures. Failure rates after massive allograft reconstructions have been reported as high as 60% at 10 years, which are associated with a multitude of biologic processes influencing the graft incorporation and functional capacity. It is unknown if mechanical failure is associated with a gradual loss of bulk material properties of the bone (strength and modulus), loss of bone mineral density, osteoclastic resorption of the allograft, unrepaired allograft microfractures or microcracks, and/or local stress concentration within the tissue. Allograft material properties, bone mineral density, microcrack prevalence, and cortical porosity were quantified in 13 failed human allograft retrievals ranging in longevity from 1 to 13 years in vivo. Nonimplanted allograft tissue (n = 27) served as the baseline for comparison. A 50% loss in strength of allograft tissue was noted after 10 years in vivo. Loss of strength was correlated with an increase in microfracture prevalence and decrease in bone mineral density within the retrieved allograft cortex. This study suggests functional failure of allograft limb salvage procedures may, in part, be attributed to degradation of the tissue's material properties, bone mineral density and prevalence of microcracks.

Grafting of massive tibial subchondral bone defects in a caprine model using beta-tricalcium phosphate versus autograft.

OBJECTIVE: This study evaluated the ability of beta-tricalcium phosphate particles (beta-TCP) and autograft (AUTO) to maintain joint surface morphology when used to supplement massive subchondral bone defects in a caprine model. DESIGN: This was a prospective, parallel arm study with 2 experimental arms and a control group. METHODS: Unilateral, 11 mm diameter, 25 mm deep cylindrical defects were created in tibial subchondral bone of anesthetized goats (n = 16) and filled with autograft or beta-tricalcium phosphate particles. The contralateral limbs served as internal controls. Goats were killed at 3 months and both tibiae harvested. Molds made of the tibial plateau surface were used to create positive casts from which medial and lateral tibial plateau surfaces of both experimental (beta-tricalcium phosphate particles, autograft) and control limbs were digitized in 3 dimensions. Mirror images of the medial condyle surface contours from the controls were superimposed onto the experimental surfaces and deviations were compared using a Student t test (alpha = 0.05). Tibiae were then cut sagittally into medial (biomechanics) and lateral (histology) halves. Compressive modulus within the defect area was assessed by indentation to 2.0 mm at 0.2 mm per second using a 6-mm diameter pin. Specimens from the lateral tibial plateau were processed for undecalcified histology and the area of bone within the defect region measured. The articular surface of 86% of the autograft and 0% of the beta-tricalcium phosphate particles group had degenerative changes, with 29% of autograft goats exhibiting large-scale plateau collapse. Mean surface deviation for autograft was significantly greater than for beta-tricalcium phosphate particles (2.19 +/- 1.49 mm versus 0.78 +/- 0.19 mm), as was maximum surface deviation (11.19 +/- 8.02 mm versus 4.39 +/- 1.33 mm) (P < 0.05). The compressive modulus within the defect area for control animals was significantly higher than the experimental groups (P < 0.05). Significantly more bone was regenerated within beta-tricalcium phosphate particle-grafted defects compared to autograft (P < 0.05). These results indicated that beta-tricalcium phosphate particles might be a useful graft material for local repair of load bearing skeletal sites such as depressed tibial plateau fractures.

An assessment of equine cartilage degeneration.

Millions of Americans suffer from osteoarthritis, a joint disease characterized by cartilage degradation and subchondral bone sclerosis. However, little is known about its pathology. It remains to be discovered which comes first in the progression of osteoarthritis: subchondral bone remodeling or cartilage degeneration. This study assessed equine cartilage degeneration, based upon measurements of cartilage thickness and Indian ink stain uptake. By gaining a greater understanding of the determining factors in cartilage degeneration, we may be able to better understand the pathomechanics of osteoarthritis. In seven horses, joint regions of interest were harvested bilaterally and isolated into planar osteochondral samples using a band saw and stored at -20 degrees C. After thawing, the samples' articular surfaces were dyed using an Indian ink stain to highlight articular cartilage degeneration. Digital images of the samples were taken before and after the staining, and mean pixel values for the pre- and post-ink images were measured using image analysis software. Reflectance Score (RS) was calculated using mean pixel values normalized between grayscale calibration standards. Articular cartilage thickness was measured at five random locations on each sample from images taken in the transverse orientation. Statistical analysis found no significant effect of limb side for either RS values or thickness, allowing data from right and left limbs to be grouped for analysis. There is a statistically significant correlation between joint region and thickness, as well as between joint region and RS; however, there was no statistical correlation between thickness and RS.

Efficacy of osteogenic protein-1 in a challenging multilevel fusion model.

STUDY DESIGN: A therapeutic study compared the influence of osteogenic protein-1 to autograft and collagen carrier in multilevel sheep spine fusions. OBJECTIVE: To evaluate the efficacy of osteogenic protein-1 compared to autograft and collagen carrier in achieving fusion in a challenging multilevel lumbar spine ovine model. SUMMARY OF BACKGROUND DATA: Bone morphogenetic proteins can successfully augment spinal fusion. To date, all the preclinical and clinical studies using bone morphogenetic proteins have evaluated single-level fusion. In practice, multiple level fusions are commonly required for various conditions, like spinal deformity. METHODS: Eighteen sheep underwent three-level spine fusion. Six sheep were treated with osteogenic protein-1 and its carrier, autograft, or with the carrier alone. Specimens were analyzed for evidence of fusion by palpation, radiographic and histologic analysis, and biomechanical testing. RESULTS: Manual palpation testing for the presence of fusion showed none of the specimens fused all three levels or fused at the lumbosacral junction. No statistically significant difference was found between the osteogenic protein-1 and autograft groups' fusion rates based on radiographic grading (P = 0.65) or biomechanical testing. Histologic analysis showed no qualitative difference in bone morphology or cellularity of fusion masses when comparing the autograft and osteogenic protein-1 specimens. CONCLUSIONS: No model before this exists that tests the efficacy of bone morphogenic proteins in as challenging an environment. Extrapolation of single-level preclinical and clinical studies with bone morphogenic proteins for use in multilevel fusion requires careful review. Autograft and osteogenic protein-1 had similar rates of fusion. A high rate of nonunion is seen with this multiple level fusion to the sacrum using autograft or osteogenic protein-1. The biologic enhancement with osteogenic protein-1 is not able to overcome this mechanically rigorous model.

Use of a resorbable sheet in iliac crest reconstruction in a sheep model.

OBJECT: Iliac crest bone graft harvesting can result in major complications, the rates of which range from approximately 6 to 8%. The objective of this study was to evaluate the postoperative regeneration of iliac crest donor defects in an animal model after harvesting a full-thickness tricortical graft. METHODS: In skeletally mature sheep, a tricortical iliac crest graft was harvested. The graft sites were allowed to heal unprotected or protected with the resorbable polylactic acid sheet material, MacroPore OS Protective Sheeting. After 6 months of healing, the sites were assessed by examination of undecalcified histological sections. Histomorphometric measurements of the original defect area, the area of new bone within the defect site, and the percentage of defect filled with new bone were quantified for both control and protected groups. In all histological sections, new bone growth within the defect sites appeared normal, with no observed excessive inflammatory cells. The developing bone tissue appeared to be remodeling normally. For the unprotected sites, the area of new bone averaged 16.3 mm2 (+/- 7.2 mm2), and the percentage of the defect area filled with bone averaged 10.7% (+/- 6.5%). In the protected sites, the area of new bone averaged 64.8 mm2 (+/- 11.6 mm2) and the percentage of the defect area filled with bone averaged 25.9% (+/- 1.6%). Both differences in area of new bone growth and percentage of defect area filled were statistically significant. Literature review has indicated that regeneration of donor site defects is desirable. CONCLUSIONS: Based on the results of the present study, MacroPore resorbable Protective Sheeting can improve bone regeneration significantly within the donor site following tricortical iliac crest graft harvesting.

Use of a resorbable sheet in iliac crest reconstruction in a sheep model.

Iliac crest bone graft harvesting can result in major complications that occur at rates of approximately 6%-8%. The objective of this study was to evaluate the postoperative regeneration of iliac crest donor defects in an animal model after harvesting a full-thickness tricortical graft. In skeletally mature sheep, a tricortical iliac crest graft was harvested. The graft sites were allowed to heal nonprotected or protected with the resorbable polylactic acid sheet material, MacroPore OS Protective Sheeting, (MacroPore Biosurgery, San Diego, Calif). After 6 months of healing, the sites were assessed by examination of undecalcified histologic sections. Histomorphometric measurements of the original defect area, the area of new bone within the defect site, and the percentage of defect filled with new bone were quantified for both control and protected groups. In all histologic sections, new bone growth within the defect sites appeared normal, with no excessive inflammatory cells observed. The developing bone tissue appeared to be remodeling normally. For the nonprotected sites, the area of new bone averaged 16.3 mm2 (+/-7.2 mm2), and the percentage of the defect area filled with bone averaged 10.7% (+/-6.5%). In the protected sites, the area of new bone averaged 64.8 mm2 (+/-11.6 mm2) and the percentage of the defect area filled with bone averaged 25.9% (+/-1.6%). Both differences in area of new bone growth and percentage of defect area filled were statistically significant. Literature review has indicated that regeneration of donor site defects is desirable. Based upon the results of the present study, MacroPore resorbable protective sheeting can improve bone regeneration significantly within the donor site following tricortical iliac crest graft harvesting.