Posterolateral Spine Research Articles

P91. Identification of Cell Surface Receptors Mediating BMP-7 Induced Osteoblast Differentiation of Primary Human Mesenchymal Stem Cells

BACKGROUND CONTEXT: Bone Morphogenetic Proteins (BMPs) are members of the TGF beta superfamily of growth factors. BMPs regulate numerous biological processes and are known to induce new bone formation in both developing and mature skeletal systems. Osteogenic protein-1 (OP-1/BMP-7) is a well studied member of this family of growth factors and is now being used clinically in revision posterolateral spine fusions and long bone non unions.The cellular responses to BMPs are mediated by specific tetrameric receptor complexes, consisting of two Type I and two Type II receptors.

A Bridging Demineralized Bone Implant Facilitates Posterolateral Lumbar Fusion in New Zealand White Rabbits

Randomized controlled animal study. Test the effectiveness of a single-strip demineralized bone matrix with hyaluronan and gelatin. Demineralized bone matrix is widely used to augment spinal fusion, however, the effect of changing the physical characteristic and carrier is not known. Demineralized bone matrix was extracted from the bones of New Zealand White rabbits, and combined with hyaluronan and gelatin to form solid strips. Forty-eight rabbits were randomized into a control and 2 experimental groups. In the control group, fusion was attempted with autograft bone. For group 2, demineralized bone matrix strips alone and for group 3, autograft and demineralized bone matrix strips were used. The fusion was assessed with manual manipulation and radiographs. The volume of the fusion mass was determined from computed tomographic images. By the manual palpation test, the fusion rates were 37.5%, 93.8%, and 100%, for groups 1-3, respectively (P < 0.05). By radiography, the control group fusion rate was 68.7% compared with 100% for the experimental groups (P < 0.05). The mean bone volumes of the fusion mass were 2142.2 +/- 318.5, 3132.9 +/- 632.1, and 4181.6 +/- 609.5 mm3 for groups 1-3, respectively (P < 0.05). The demineralized bone matrix-gel strip was able to function as both a bone-graft enhancer and a bone graft substitute in rabbit posterolateral spine fusion.

Posterolateral lumbar spine fusion with INFUSE bone graft

INFUSE has been proven effective in conjunction with threaded cages and bone dowels for single-level anterior lumbar interbody fusion (ALIF). The published experience with posterolateral fusion, although encouraging, utilizes a significantly higher dose and concentration of recombinant human bone morphogenic protein-2 (rhBMP-2) and a different carrier than the commercially available INFUSE. To present an assessment of fusion rate for posterolateral spine fusion with INFUSE Bone Graft. Retrospective review of patients treated using INFUSE in posterolateral spine fusion in a single institution. 91 patients with minimum 2-year follow-up who underwent posterolateral spine fusion using INFUSE as an iliac crest bone graft (ICBG) substitute. Fusion rate based on fine-cut computed tomographic (CT) scans with sagittal and coronal reconstructions. Fusion was performed using one large INFUSE kit (12 mg rhBMP-2, 1.5 mg/mL), which was prepared according to the manufacturer's instructions. The INFUSE sponge was wrapped around the local bone or graft extender and placed over the decorticated surfaces in the lateral gutters. Postoperative CT scans with reconstructions were reviewed by two independent orthopedic spine surgeons. CT scans of a comparison group of 35 patients who underwent primary single-level posterolateral fusion with ICBG were also reviewed. The overall group had a mean 4.38 CT fusion grade and a 6.6% nonunion rate. Primary one-level fusion cases (n=48) had a mean 4.42 fusion grade a 4.2% nonunion rate. Primary multilevel fusions (n=27) had a mean 4.65 CT grade and no nonunions detected. Assessment of the 35 primary one-level ICBG control cases demonstrated a mean CT grade of 4.35 and a nonunion rate of 11.4%. In the 16 cases of revision for prior nonunion, mean CT grade was 3.81 and 4 subjects had nonunions. Additional subgroup analysis showed that smokers (n=14) had a mean 4.32 CT grade with no nonunions. Men had a mean 4.04 CT grade and an 11.1% nonunion rate compared with a mean 4.61 CT grade and 3.6% nonunion rate in women. This difference was statistically significant (p=.036). No significant differences in fusion rate were observed based upon the specific graft extender used (p=.200). Posterolateral spine fusion involves a more difficult healing environment with a limited surface for healing, a gap between transverse processes and the milieu of distractive forces. Historically, only ICBG has been able to overcome these challenges and reliably generate a successful posterolateral lumbar spine fusion. In contrast to prior studies, clinically available INFUSE delivers only 12 mg rhBMP-2 at a concentration of 1.5 mg/mL. Despite the lower dose and concentration of rhBMP-2, this study suggests that fusion success with INFUSE is equivalent to ICBG for posterolateral spine fusion. As with ICBG, development of solid fusion or nonunion is a multifactorial process. The use of INFUSE is not a substitute for proper surgical technique or optimization of patient-related risk factors. Additional studies are needed to determine the incremental benefit of a greater rhBMP-2 dose or use of alternative carriers for posterolateral fusion. Finally, correlation between radiographic findings and clinical outcomes, and a cost-benefit analysis are needed. Despite these issues, this study presents compelling evidence that commercially available INFUSE is an effective ICBG substitute for one- and two-level posterolateral instrumented spine fusion.

Posterolateral intertransverse lumbar fusion in a mouse model: surgical anatomy and operative technique

Animal models are frequently used for studying the effect of bone graft substitutes or allogeneic materials on osterolateral lumbar fusion. Transgenic technology in the mouse provides a unique opportunity to further understand the biology of spine fusion. To describe pertinent lumbar spine anatomy and formulate a surgical protocol for posterolateral fusion in the mouse model. Diagnostic model: development of an animal model for biologic evaluation of posterolateral spine fusion. Ten mice were killed to study relevant lumbar spine anatomy and develop a protocol for lumbar spine fusion. The L4-L6 fusion protocol was validated in 46 mice for ease of exposure, preparation of the posterolateral fusion bed, introduction of bone inductive agents, and perioperative care. Anatomy and surgical technique for posterolateral intertransverse lumbar fusion in the mouse model are described. A paraspinal approach allows exposure of the transverse processes, decortication, and graft placement at the L4-L6 intertransverse fusion site. Decortication alone did not result in fusion, whereas the use of bone graft resulted in satisfactory fusion rates. Perioperative morbidity and mortality rates were low. The mouse posterolateral lumbar spine fusion model is reproducible, inexpensive, and has low complication rates. Knowledge of the relevant anatomy and adherence to a well-defined surgical protocol provides a reliable and reproducible experimental spine fusion model.

Gene Expression for Type-Specific Collagens in Osteogenic Protein-1 (rhBMP-7)-Induced Lumbar Intertransverse Process Fusion in Rabbits

To elucidate the molecular mechanism by which osteogenic protein (OP)-1 induces posterolateral lumbar spine fusion (PLF), we analyzed the process of OP-1-induced PLF in an established rabbit model by means of in situ hybridization (ISH). Intertransverse process lumbar fusions were performed at L5-L6 in rabbits using OP-1 (n = 18) or carrier alone (n = 9). The vertebrae were harvested at 2, 4, and 6 weeks postsurgery, and fusion masses evaluated using radiography, histology and ISH. In the OP-1 group, a contiguous fusion mass bridged the L5 and L6 transverse processes by 6 weeks postsurgery. At 2 weeks, collagen types II and X were expressed both near the transverse process and at the central portion of the intertransverse process area, corresponding to abundant, multifocal cartilage. After 4 weeks, endochondral ossification progressed from the transverse process toward the central portion. At 6 weeks, genes for collagen types II and X were restricted at the front of endochondral ossification. In carrier-alone group, no fusion mass was formed. The results demonstrate that OP-1 vigorously stimulates cells and induces chondrogenesis in the early phase of PLF. In the late phase, however, endochondral bone formation progressing toward the central portion became the major process, as was observed in PLF with autograft. In OP-1-induced PLF, therefore, osteogenic action of OP-1 seems to be quite intensive in the early phase but not in the late phase.

Assessment of Malunion in Spinal Fusion

Pseudarthrosis, or non-union, is a relatively common and occasionally difficult problem for the spine surgeon to treat. It often appears weeks of months after the initial surgery, and presents with pain or neurologic deterioration after a period of improvement. Pseudarthrosis is defined as the persistence of a motion segment after attempted bony fusion, either at the interbody space or the posterolateral spine. The causes of non-union can be difficult to ascertain and are often multifactorial; they can include use of nicotine or certain pharmacologic agents; patient co-morbidities; and osteoinductive properties of the bone used for fusion. Pseudarthrosis may be difficult to diagnose, and often requires the use of several radiologic studies, including reformatted CT scan. The gold standard for determining if pseudarthrosis exists, open exploration, is usually unfeasible. Treatment options for symptomatic pseudarthrosis include operative intervention, either posteriorly or anteriorly (or both), and fusion rates can be enhanced with the use of internal fixation, bone morphogenic protein, and electrical stimulation. The best treatment for pseudarthrosis remains prevention. This review discusses the pathophysiology, diagnosis, prevention and treatment of spinal pseudarthrosis.

P129. The efficacy of kryptonite in a rat model of posterolateral spine fusion

BACKGROUND CONTEXT: Although autogenous bone graft is the most widely used source of graft material for spinal fusion, its associated morbidity and limitations has motivated the search for bone graft alternatives. One such alternative is bioabsorbable-calcified triglyceride, or Kryptonite. This triglyceride bone cement is created by combining two fatty acids (isocyanate and poliol) with calcium carbonate, which interact to form an odorless, exothermic calcified triglyceride. Once implanted, this compound is metabolized, resorbed, and replaced with bone.

The Use of Cultured Bone Marrow Cells in Type I Collagen Gel and Porous Hydroxyapatite for Posterolateral Lumbar Spine Fusion

Posterolateral lumbar transverse process fusion was completed using the cultured bone marrow cells in type I collagen gel and porous hydroxyapatite. To compare the efficacy of cultured bone marrow cells with that of bone morphogenetic protein (BMP) as a graft alternative to autologous bone for posterolateral spine fusion. The clinical application of BMP for spinal fusion may be limited by high dose and cost. Recently, mesenchymal stem cells have been studied in various fields because of their capability to differentiate into various cells, including those in the osteogenic lineage. Thirty adult rabbits were used. Each underwent single-level, bilateral, posterolateral intertransverse process fusions at L4-L5. The animals were divided into 4 groups, each according to the material implanted: (1) autologous bone (autograft, n = 9); (2) porous hydroxyapatite (HA) particles and type I collagen sheet with 100 microg rhBMP-2 (BMP-HA, n = 7); (3) bone marrow cells (1 x 10(6) cells/mL, low-marrow-HA, n = 7); and (4) bone marrow cells (1 x 10(8) cells/mL, high-marrow-HA, n = 7). Before implantation for groups 3 and 4, fresh bone marrow cells from the iliac crest of each animal were cultured in a standard medium for 2 weeks. For one additional week, the marrow cells were cultured in 10(-8) M dexamethasone, type I collagen gel, and HA. Animals were euthanized 6 weeks after surgery. Spinal fusions were evaluated by radiograph, manual palpation, and histology. The fusion rates were 4 of 7 in the autograft group, 7 of 7 in the BMP-HA group, 0 of 7 in the low-marrow-HA group, and 5 of 7 in the high-marrow-HA group. The histology in the BMP-HA and high-marrow-HA groups showed that grafted HA fragments were connected with mature new bone. The pores of HA fragments were filled up with bone matrix. In the low-marrow-HA group, fibrous tissue was predominant in the grafted fragments. This study shows that the cultured bone marrow cells can act as a substitute for autograft or BMP in spine fusion. The current formulation may yield improved fusion success and better quality of fusion bone as compared to autograft.

Lower Dose of rhBMP-2 Achieves Spine Fusion When Combined With an Osteoconductive Bulking Agent in Non-human Primates

A non-human primate lumbar intertransverse process arthrodesis model was used to evaluate the effectiveness of different formulations of recombinant human bone morphogenetic protein-2 (rhBMP-2) to induce consistent bone formation. To determine if the combination of rhBMP-2/absorbable collagen sponge (ACS) wrapped around a bulking agent, consisting of a biphasic calcium phosphate/collagen composite, could achieve posterolateral spine fusion with a dose of rhBMP-2 (3.0 mg/side) that previously failed to induce posterolateral fusion in rhesus monkeys with other carriers. Successful bone induction in both human and non-human primates has required higher concentrations of BMP than were required in lower order models. The Food and Drug Administration approved concentration of rhBMP-2 for interbody fusion (1.5 mg/mL) when delivered on the ACS alone without a bulking agent (doses 3-9 mg/side) has failed to induce clinically relevant amounts of bone formation in a posterolateral spine fusion model in rhesus monkeys. Previously, a higher concentration of 2.0 mg/mL of rhBMP-2 delivered on stacked sheets of a biphasic calcium phosphate ceramic/collagen compression resistant matrix (CRM) was required to achieve fusion in the rhesus monkey and was the basis for this study (doses of 6-12 mg/side). Nine skeletally mature, rhesus macaque monkeys underwent single level posterolateral arthrodesis at L4-L5. Two different rhBMP-2 doses were evaluated in 3 delivery configurations. The first 3 monkeys received 10 mg/side (2.5 mL at 4.0 mg/mL) of rhBMP-2 loadeddirectly onto a CRM carrier (15% hydroxyapatite/85%beta-tricalcium phosphate ceramic/collagen matrix), resulting in a final concentration of 2.0 mg/mL. The second 3 monkeys received 3 mg/side (2.0 mL at 1.5 mg/mL) of rhBMP-2 loaded directly on the CRM carrier, resulting in a 0.6 mg/mL final concentration. Three additional monkeys also received the 3 mg/side (2.0 mL at 1.5 mg/mL) of rhBMP-2 delivered on an ACS, which was then wrapped around the dry CRM matrix used as a bulking agent, yielding a 1.5 mg/mL final concentration of rhBMP-2 on the sponge wrapped around the bulking agent. The monkeys were euthanized at 24 weeks after surgery. Manual palpation, plain radiographs, computerized tomography, and nondecalcified histology were used to evaluate fusion in a blinded fashion. The 3 monkeys with 10 mg rhBMP-2 placed directly on the CRM carrier (2.0 mg/mL final concentration) achieved solid fusion. The 3 monkeys that underwent fusion with 3 mg of rhBMP-2 placed directly on the CRM carrier (0.6 mg/mL final concentration) failed to achieve fusion. In contrast, the 3 monkeys that underwent fusion with the same 3 mg dose of rhBMP-2 dispensed only on an ACS that was wrapped around the CRM achieved solid bilateral fusion. This study shows the importance of carrier optimization and final implant protein concentration for the successful delivery of rhBMP-2. By combining the properties of the ACS with the CRM, the required dosage of rhBMP-2 was diminished by more than 3-fold in the non-human primate model. This finding suggests that the currently available concentration of rhBMP-2 (1.5 mg/mL) could be successful for achieving posterolateral spine fusion when combined with an osteoconductive bulking agent that can support the induced new bone formation.

Genetically Modified Human Derived Bone Marrow Cells for Posterolateral Lumbar Spine Fusion in Athymic Rats

This experimental study consisted of four groups. In Group I, 8 animals were implanted with a collagen sponge containing 5 million human-derived bone marrow cells (HBMCs) infected with the BMP-2-containing adenovirus. In Group II, 5 animals were implanted with a collagen sponge containing 5 million HBMCs, which had been infected with an adenovirus containing the cDNA for the lacZ gene. In Group III, 5 animals were implanted with a collagen sponge containing 5 million uninfected HBMCs. In Group IV, 5 animals were implanted with a collagen sponge alone. All animals were killed at 12 weeks. The purpose of this study was to assess the ability of HBMCs infected with a BMP-2 adenovirus to induce a posterolateral spine in an athymic rat model. High pseudarthrosis rates after attempted spinal arthrodesis as well as the morbidity associated with iliac crest bone graft harvest make alternative bone graft materials desirable. Alternatives include allograft bone, demineralized bone matrices, and more recently, recombinant bone morphogenetic proteins. However, high doses of the recombinant protein are required, and a single dose of recombinant protein may not induce a sufficient osteoinductive response in all patients. An alternative strategy is to genetically manipulate cells to overexpress a bone morphogenetic protein. HBMCs grown in culture were infected with an adenovirus containing the cDNA for BMP-2 (AdBMP-2) or the gene for beta-galactosidase (AdlacZ). Cells were implanted on the decorticated transverse processes of L4-L5 as previously described. Rats were assessed by radiographs at 4-week intervals and were killed at 12 weeks. After death, spines were explanted and assessed by manual palpation and histologic analysis. Eight of eight rats implanted with AdBMP-2-infected HBMCs (Group I) were fused by radiographic evaluation and manual palpation at 12 weeks. Control groups consisted of: 5 rats implanted with the collagen sponge alone (Group II), 5 rats implanted with a collagen sponge and 5 million uninfected HBMCs (Group III), and 5 rats implanted with HBMCs infected with AdlacZ (Group IV). None of the rats in the control groups (0 of 15) developed a fusion at L4-L5. This study demonstrates that human-derived bone marrow cells can be infected with a BMP-2-containing adenovirus and produce sufficient bone in vivo to fuse the lumbar spine.

Evaluation of HealosMP52 Osteoinductive Bone Graft for Instrumented Lumbar Intertransverse Process Fusion in Sheep

HealosMP52 was evaluated in a sheep model of instrumented lumbar intertransverse process spine fusion and compared to autogenous bone graft. To determine the long-term efficacy and safety of HealosMP52 as a bone graft substitute in posterolateral instrumented spinal fusion. Although the standard intertransverse fusion method employs autogenous iliac crest bone, autograft has certain limitations. HealosMP52, an osteoinductive bone graft material, can facilitate noninstrumented posterolateral spine fusion in rabbits and nonhuman primates, but the long-term outcome of such fusions has not been evaluated. Eleven skeletally mature, female sheep were instrumented with pedicle screws and rods at L2-L3 and L5-L6. Each animal was treated with autograft bone at one fusion level and HealosMP52 at the other. At 6 and 12 months after surgery, bone formation was measured on contact microradiographs and by backscattered electron imaging. Bone core biopsies taken from 6-month and 12-month specimens were evaluated histologically for pathology indicative of osteosarcoma. Grossly, all autograft- and HealosMP52-treated levels showed stable fusions at 6 and 12 months. HealosMP52 and autograft treatments resulted in equivalent mean percent bone volumes within fusion bodies; similar values were observed at 6 and 12 months. Fusion bodies contained cortical and trabecular bone with osteoid seams and fatty marrow, and fusion masses showed maturation from 6 to 12 months. HealosMP52 treatment was not associated with implant migration, ectopic bone formation, or pathologic abnormality. No histologic evidence of osteosarcoma was seen on bone core biopsies. This long-term assessment of the use of HealosMP52 in posterolateral instrumented spine fusion indicates that HealosMP52 possesses safe and efficacious bone grafting properties and can potentially serve as anosteoinductive alternative to autograft bone.

Combined Bone Morphogenetic Protein-2 and −7 Gene Transfer Enhances Osteoblastic Differentiation and Spine Fusion in a Rodent Model

To enhance the osteogenic activity of BMP, combination BMP2 and BMP7 gene transfer was performed. This approach led to a significant increase in osteoblastic differentiation of mesenchymal precursors compared with single BMP gene transfer in vitro. When tested in 78 rats, combination gene transfer enhanced mechanically stable spine fusion and bone formation rate versus single BMP gene transfer. Although clinical bone morphogenetic protein (BMP) therapy is effective, required doses are very high. Previous studies have suggested that the co-expression of two different BMP genes can result in the production of heterodimeric BMPs that may be more potent than homodimers. In this study, combined BMP2 and BMP7 gene transfer was performed to test whether this approach improves osteoblastic differentiation and bone formation compared with single BMP gene transfer. A producer cell (A549) was co-transfected with adenovirus vectors encoding BMP2 (AdBMP2) and BMP7 (AdBMP7) or, as controls, each vector alone, AdNull (with no transgene) or no virus. Supernatants were compared for their ability to stimulate osteoblastic differentiation of C2C12 myoblasts and MC3T3-E1 pre-osteoblasts. In a rat posterolateral spine fusion model, co-administration of AdBMP2 and AdBMP7 was compared with treatment with each vector alone, AdNull or no virus in 78 rats. The spines were assessed 8 weeks after surgery for radiographic and mechanical fusion, bone formation, and mineralization. BMP2 and BMP7 were co-precipitated from supernatants of cells co-transfected with AdBMP2 and AdBMP7, indicating the presence of BMP2/7 heterodimer. Supernatants of co-transfected cells containing relatively low doses (7-140 ng/ml) of BMPs induced osteocalcin expression and alkaline phosphatase activity in both C2C12 and MC3T3-E1 cells, that were up to 6- and 40-fold higher, respectively, than levels induced by maximal doses (200-1000 ng/ml) of either BMP2 or BMP7 alone. In the spine fusion model, co-administration of AdBMP2 and AdBMP7 resulted in a significantly greater number of mechanically stable fusions and also 2-fold higher mineralization rate and bone volume in the fusion mass versus single BMP gene transfer (p < 0.02, all comparisons). Combined BMP2 and BMP7 gene transfer is significantly more effective in inducing osteoblastic differentiation and spine fusion than individual BMP gene transfer.

A Pilot Study Evaluating the Safety and Efficacy of OP-1 Putty (rhBMP-7) as a Replacement for Iliac Crest Autograft in Posterolateral Lumbar Arthrodesis for Degenerative Spondylolisthesis

A prospective, randomized, controlled, multicenter clinical study was conducted. To compare the clinical and radiographic outcomes of patients treated with OP-1 (BMP-7) Putty to autogenous iliac crest bone graft for one-level uninstrumented posterolateral fusion of the lumbar spine following decompressive laminectomy for the treatment of symptomatic degenerative spondylolisthesis with spinal stenosis. Preclinical studies have demonstrated that osteoinductive recombinant human osteogenic protein 1 in the form of OP-1 Putty is successful at achieving a posterolateral fusion in rabbits and dogs without any significant safety concerns. Thirty-six patients with degenerative lumbar spondylolisthesis and symptoms of neurogenic claudication were randomized (2:1) to either OP-1 Putty (3.5 mg of OP-1 per side) or autogenous iliac crest bone graft for one-level uninstrumented posterolateral fusion following a decompressive laminectomy. Enrollment in the study was complete when 24 OP-1 Putty patients and 12 autograft patients had been randomized and treated. A patient administered Oswestry scale and SF-36 scale were used to determine clinical outcomes. Independent, blinded neuroradiologists reviewed both static and dynamic radiographs to determine fusion status. At the 1-year follow-up, 32 patients were available for clinical analysis and 29 patients were available for radiographic review. Clinical success as measured on the Oswestry scale was achieved by 18 of 21 (86%) OP-1 Putty patients and 8 of 11 (73%) autograft patients. SF-36 pain index scores showed similar results. Fourteen of 19 (74%) OP-1 Putty patients and 6 of 10 (60%) autograft patients achieved a successful posterolateral fusion fulfilling all fusion criteria. Of the 29 evaluable patients, 15 were both clinical and radiographic successes, 5 were radiographic successes but were clinical failures, 1 patient was both a radiographic and clinical failure, and 8 patients were radiographic failures but were clinical successes. No systemic toxicity, ectopic bone formation, recurrent stenosis, or other adverse events related to the OP-1 Putty implant were observed. Although the posterolateral spine is a challenging fusion environment in patients with degenerative spondylolisthesis, successful radiographic fusion was obtained using OP-1 Putty at a rate that was similar to autograft given the number of patients in this study. Importantly, there were no apparent adverse consequences related to the use of the OP-1 Putty implant in this patient population.

70. Prospective randomized evaluation of Healos/Bone Marrow Aspirate vs autologous iliac crest bone in instrumented posterolateral lumbar fusion

BACKGROUND CONTEXT: Autologous iliac crest bone graft is the “gold standard” in instrumented posterolateral spine arthrodesis. Recent reviews have indicated a significant complication rate as well as on-going pain related to the harvest of such bone. Healos is an osteoconductive bovine type I collagen matrix uniformly coated with hydroxyapatite. The combination of Healos and bone marrow aspirate have shown high rates of fusion in both animal and human studies.

P53. Effects of capacitive coupling electrical stimulation on expression of growth factors in a rabbit posterolateral spine fusion model

back pain in 2000. Among individual drugs, ibuprofen and naproxen accounted for most of the prescriptions for traditional NSAIDs (60%), whereas two-thirds of the prescriptions for muscle relaxants were attributable to cyclobenzaprine, carisoprodol and methocarbamol. Prescription of COX-2 inhibitors or muscle relaxants demonstrated wide variations across different regions. Several individual characteristics including age, race and educational level were associated with the prescription of some of the medications. CONCLUSIONS: Neither traditional NSAIDs, nor COX-2 inhibitors, nor muscle relaxants dominated prescriptions for back pain. However, a small number of individual drugs were attributable to most of the prescriptions for traditional NSAIDs or muscle relaxants. The prescription of some of the medications demonstrated wide variations across different regions or different racial and educational groups. More studies are needed to understand why the variations occurred and how to standardize the prescriptions. Head-to-head comparisons of the relative costs and effectiveness among different categories of the medications, especially those drugs that have been most widely prescribed should also be warranted. DISCLOSURES: No disclosures. CONFLICT OF INTEREST: No Conflicts.

99. Gene therapy utilizing stem cells from human fat in a rat posterolateral spine fusion model

99. Gene therapy utilizing stem cells from human fat in a rat posterolateral spine fusion model

Evidence of osteoinduction by Grafton demineralized bone matrix in nonhuman primate spinal fusion.

A nonhuman primate posterolateral lumbar intertransverse process arthrodesis model was used to evaluate osteoinductive bone graft materials. To test two new formulations of Grafton demineralized bone matrix (Flex and Matrix) for evidence of osteoinduction and their potential efficacy as an extender or enhancer for autogenous bone in a previously validated nonhuman primate posterolateral lumbar fusion model. Whereas several demineralized bone matrix formulations have been shown to be variably osteoinductive in rodent ectopic bone assays and rabbit spine applications, few have demonstrated efficacy in higher species and in more challenging applications such as posterolateral spine fusion. The authors are not aware of any published studies describing any demineralized bone matrix that has been tested in a nonhuman primate posterolateral spine fusion model. After approval by the institutional animal care and use committee, eight skeletally mature rhesus macaques underwent single-level posterolateral arthrodesis. In four animals, autograft (4 g/side) was implanted with a piece of human Grafton Flex demineralized bone matrix. In the other four animals, rhesus Grafton Matrix demineralized bone matrix, a new and more porous formulation of Flex, was implanted with autograft (4 g) on one side of the spine, and Matrix with half the amount of autograft (2 g) was implanted on the opposite side. Radiographs were taken at intervals until the animals were killed at 24 weeks. Spinal fusion was evaluated by manual palpation (status was fused or not fused), and computed tomography was done to visualize the amount of bone formation. Fusion was ascertained by palpation in two of four monkeys receiving Flex with autograft and in three of four monkeys receiving Matrix with autograft. Evidence of osteoinduction was seen in all four monkeys on the Matrix with 4 g autograft side, which had larger fusion masses than in the other treatments. Histologic examination showed that the bone formed was normal. The rhesus Matrix formulation performed better than the human Flex. Evidence of osteoinduction was seen in all four monkeys that received Matrix, which improved the fusion success of autograft. This alone suggested that it might play a role as a graft enhancer, not merely as a graft extender. Human studies are warranted.

Augmentation of Rabbit Posterolateral Spondylodesis Using a Novel Demineralized Bone Matrix-Hyaluronan Putty

Posterolateral spinal fusion with allogeneic demineralized bone graft-hyaluronan putty in addition to autogenous iliac crest bone graft in a rabbit model. To determine the potential efficacy of demineralized bone graft-hyaluronan putty as a bone graft enhancer. Autograft bone is the material of choice for posterolateral lumbar intertransverse process fusion. Bone graft alternatives such as demineralized bone matrices that can be used as graft extenders, enhancers, or substitutes continue to be developed. One hundred New Zealand white rabbits underwent bilateral posterolateral spinal fusion with autogenous iliac crest bone graft or bone graft with allogeneic rabbit demineralized bone graft-hyaluronan putty. The rabbits were killed 9 weeks later, and the lumbar spines were removed. Manual manipulation and fine detail radiography were used to assess spinal fusion, and computed tomographic images were used to quantify the volume of the fusion mass. In comparison with autograft bone alone, the fusion rates were greater when demineralized bone graft-hyaluronan putty was used as an adjunct to autogenous bone. Furthermore, the radiographic fusion rate was greater when demineralized bone graft-hyaluronan putty was used in a 2:1 ratio to autograft bone in comparison with a 1:1 ratio (P = 0.001). The addition of demineralized bone graft-hyaluronan putty to autograft bone was found to increase mineralized bone volume in a ratio-dependent manner (P < 0.05). Allogeneic demineralized bone matrix-hyaluronan putty enhances rabbit posterolateral spine fusion when used as an adjunct to autogenous bone graft. This new formulation of demineralized bone matrix may facilitate greater bone formation and successful fusion.

48. Effects of direct current electrical stimulation on expression of BMP 2, 4, 6, 7, bFGF, VEGF, TGF-β, ALK2 and ALK3 in a rabbit posterolateral spine fusion model

HYPOTHESIS: Direct current (DC) electrical stimulation as an adjunct treatment in spinal surgery has shown increased fusion rates when compared to autograft alone, yet the biology of such treatment is not fully understood. This study tested the possibility that DC stimulation at the surgery site would increase expression of genes related to bone formation relative to expression in autograft alone.

Simple carrier matrix modifications can enhance delivery of recombinant human bone morphogenetic protein-2 for posterolateral spine fusion.

A nonhuman primate lumbar intertransverse process arthrodesis model was used to evaluate modifications to a plain collagen sponge to deliver recombinant human bone morphogenetic protein-2 (rhBMP-2). To evaluate the feasibility of enhancing the delivery of rhBMP-2 with the established collagen sponge carrier by adding biphasic ceramic phosphate (BCP) granules (15% hydroxyapatite, 85% tricalcium phosphate) or allograft chips to provide compression resistance for posterolateral spine arthrodesis. Recombinant human bone morphogenetic protein-2 was successfully delivered with a resorbable collagen sponge in a rabbit intertransverse process fusion model. Success in nonhuman primates required a higher dose (6-9 mg) of rhBMP-2 and a more compression-resistant matrix (ceramic) than plain collagen. The limitation of the ceramic carrier was its radiopacity, which made radiographic detection of new bone formation difficult. Nine adult rhesus monkeys underwent bilateral posterolateral intertransverse process arthrodesis at L4-L5. The animals were divided into three groups (n = 3 each) based on the graft material implanted: 1) autogenous iliac crest bone (5 cm3/side); 2) collagen sponge and 15:85 BCP granules loaded with rhBMP-2 (3 mg/side); and, 3) collagen sponge and allograft chips loaded with rhBMP-2 (3 mg/side). The monkeys were killed 24 weeks after surgery. Inspection, manual palpation, radiography, computed tomographic scans, and histology were used to assess fusion. All six monkeys with rhBMP-2 delivered in the collagen/15:85 BCP carrier and the collagen/allograft chips carrier achieved solid spine fusions, whereas only one of three animals fused with autogenous bone graft. Histologic analysis of the bone induced by rhBMP-2 showed normal trabecular bone and bone marrow elements. The addition of either 15:85 BCP granules or allograft bone chips to the existing resorbable collagen sponge matrix enhanced delivery of rhBMP-2 in the posterolateral spine. The combination matrices were more compression resistant and had improved radiographic resorption properties that permitted easy radiographic visualization of new bone. In addition, a lower dose of rhBMP-2 (3 mg/side) was successful compared with the dose previously used with the plain collagen sponge (6 mg/side).