Posterolateral Intertransverse Process Research Articles

A NOVEL SELECTIVE PROSTAGLANDIN-RECEPTOR EP4-AGONIST PROMOTES SPINAL FUSION IN RATS

A novel EP4 selective agonist (KMN-159) was developed [1] and has been proven that it can act as an osteopromotive factor to repair critical-size femoral bone defects in rats at a dose-dependent manner [2]. Based on its osteopromotive properties, we hypothesized that KMN-159 could also aid in bone formation for spinal fusion. Therefore, the aim of this study was to investigate its spinal fusion effect in a dorsolateral spinal fusion model in rats. This study was performed on 192, 10-week-old male Wistar rats. The rats were randomized into 8 groups (n = 12 per group): 1) SHAM (negative control), 2) MCM (scaffold only), 3) MCM + 20 µg BMP-2 (positive control), 4-8) MCM + 0.2, 2, 20, 200 or 2000 µg KMN-159. A posterolateral intertransverse process spinal fusion at L4 to L5 was performed bilaterally by implanting group dependent scaffolds (see above) or left empty in the SHAM group (protocol no. 25-5131/474/38). Animals were euthanized after 3 weeks and 6 weeks for µCT and biomechanical testing analysis. The results showed that KMN-159 promoted new bone formation in a dose-dependent manner at 3 weeks and 6 weeks as verified by µCT. The biomechanical testing showed that the dose of 20, 200 and 2000 µg KMN-159 groups obtained comparable strength with BMP-2 group, which higher than SHAM, MCM and lower doses of 0.2 and 2 µg KMN-159 groups. In conclusion, KMN-159 could be a potential replacement of BMP-2 as a novel osteopromotive factor for spinal fusion.Acknowledgements: We are grateful to Ulrike Heide, Anna-Maria Placht (assistance with surgeries) as well as Suzanne Manthey & Annett Wenke (histology).

Creation and preclinical evaluation of a novel mussel-inspired, biomimetic, bioactive bone graft scaffold: direct comparison with Infuse bone graft using a rat model of spinal fusion.

Infuse bone graft is a widely used osteoinductive adjuvant; however, the simple collagen sponge scaffold used in the implant has minimal inherent osteoinductive properties and poorly controls the delivery of the adsorbed recombinant human bone morphogenetic protein-2 (rhBMP-2). In this study, the authors sought to create a novel bone graft substitute material that overcomes the limitations of Infuse and compare the ability of this material with that of Infuse to facilitate union following spine surgery in a clinically translatable rat model of spinal fusion. The authors created a polydopamine (PDA)-infused, porous, homogeneously dispersed solid mixture of extracellular matrix and calcium phosphates (BioMim-PDA) and then compared the efficacy of this material directly with Infuse in the setting of different concentrations of rhBMP-2 using a rat model of spinal fusion. Sixty male Sprague Dawley rats were randomly assigned to each of six equal groups: 1) collagen + 0.2 µg rhBMP-2/side, 2) BioMim-PDA + 0.2 µg rhBMP-2/side, 3) collagen + 2.0 µg rhBMP-2/side, 4) BioMim-PDA + 2.0 μg rhBMP-2/side, 5) collagen + 20 µg rhBMP-2/side, and 6) BioMim-PDA + 20 µg rhBMP-2/side. All animals underwent posterolateral intertransverse process fusion at L4-5 using the assigned bone graft. Animals were euthanized 8 weeks postoperatively, and their lumbar spines were analyzed via microcomputed tomography (µCT) and histology. Spinal fusion was defined as continuous bridging bone bilaterally across the fusion site evaluated via µCT. The fusion rate was 100% in all groups except group 1 (70%) and group 4 (90%). Use of BioMim-PDA with 0.2 µg rhBMP-2 led to significantly greater results for bone volume (BV), percentage BV, and trabecular number, as well as significantly smaller trabecular separation, compared with the use of the collagen sponge with 2.0 µg rhBMP-2. The same results were observed when the use of BioMim-PDA with 2.0 µg rhBMP-2 was compared with the use of the collagen sponge with 20 µg rhBMP-2. Implantation of rhBMP-2-adsorbed BioMim-PDA scaffolds resulted in BV and bone quality superior to that afforded by treatment with rhBMP-2 concentrations 10-fold higher implanted on a conventional collagen sponge. Using BioMim-PDA (vs a collagen sponge) for rhBMP-2 delivery could significantly lower the amount of rhBMP-2 required for successful bone grafting clinically, improving device safety and decreasing costs.

P1. Efficacy of a calcium phosphate collagen matrix bone graft with needle-shaped submicron surface topography in a rabbit posterolateral lumbar spine fusion model

<h3>BACKGROUND CONTEXT</h3> Synthetic bone grafts are a promising alternative for the treatment of degenerative spine conditions and are available without the added drawbacks associated autografts including limited availability and donor-site morbidity. Of these synthetic materials, calcium phosphate (CaP) with a characteristic needle-shaped submicron surface topography (BCP<µm) has attracted much attention due to its unique bone-forming ability which is essential for repairing critical-size bone defects such as those found in the posterolateral spine [1,2]. Previous in vitro and ex-vivo data presented by van Dijk LA [3] and Yuan H [4] at NASS, 2020 demonstrated that these specific surface characteristics drive a favorable response from the innate immune system. <h3>PURPOSE</h3> This study was to evaluate the efficacy of a novel synthetic bone graft using a clinically-relevant "Boden" rabbit posterolateral spine fusion model. The synthetic bone graft comprised of CaP granules with a needle-shaped submicron surface structure (BCP<µm) embedded in a highly porous and fibrillar bovine type I collagen matrix (col-BCP<µm). The fibrillar collagen was introduced as a carrier, designed to facilitate admixing of the graft with biological fluids perioperatively and to minimize graft migration postoperatively. <h3>STUDY DESIGN/SETTING</h3> The novel synthetic bone graft was first mixed in a 1:1 volume ratio with autologous bone (ABG). Subsequently, grafts were implanted in the rabbit posterolateral spine environment for 3 days, 6, 9 and 12 weeks. An autologous bone graft group was used as a positive control in this study. Outcomes for the test article were additionally compared to historical data for the same material (BCP<µm) implanted alone, without the fibrillar collagen carrier. <h3>OUTCOME MEASURES</h3> Spinal fusion was determined by manual palpation (MP), X-ray and µCT imaging (based on the Lenke scale), mechanical testing (range of motion testing at 12 weeks only) and histology (decalcified and undecalcified). <h3>METHODS</h3> Forty skeletally mature New Zealand white rabbits were divided into two groups and underwent single-level bilateral posterolateral intertransverse process spine arthrodesis at L4-5. Animals were euthanized after 3 days, 6, 9 and 12 weeks. <h3>RESULTS</h3> Based on all endpoints, the test group and positive control group showed an equivalent and gradual progression in bone formation and implant resorption over time, leading to mature fusion masses at 12 weeks. No adverse reactions were observed in any of the groups. Bilateral fusion rates by MP were consistent with the literature (0-40% at 6 weeks; 60-70% at 12 weeks) and similar trends were seen by histological and radiographic assessment (both X-ray and µCT). Flexion-extension and lateral bending results at 12 weeks demonstrated fusion of the treated level consistent with the MP results. The results observed for the test group were consistent with those seen historically for the same material implanted without the fibrillar collagen carrier. <h3>CONCLUSIONS</h3> These results show that BCP with needle-shaped submicron-scale surface topography (BCP<µm) combined with a fibrillar collagen matrix (col-BCP<µm) could achieve equivalent fusion rates to ABG when used as a bone graft extender in a clinically relevant single level rabbit intertransverse process posterolateral lumbar fusion model. Over time, a progression in bone healing, graft resorption and remodeling were seen with all experimental endpoints, with equivalency between the groups. The results for col-BCP<µm were consistent with those previously seen for BCP<µm alone, confirming that the fibrillar collagen carrier improved handling properties but was not detrimental to the bone-forming capacity of the BCP<µm. <h3>FDA DEVICE/DRUG STATUS</h3> Col-BCP<µm (Investigational/Not approved)

Comparing the efficacy of adipose-derived and bone marrow-derived cells in a rat model of posterolateral lumbar fusion.

Although bone marrow-derived mesenchymal stem cells (BMCs) have been widely used in spinal fusion procedures, adipose-derived stem cells (ASCs) offer a number of advantages as an alternative clinical cell source. This study directly compares the efficacy of ASCs and BMCs from the same donor animals to achieve successful fusion when combined with a clinical-grade bone graft substitute in a rat lumbar fusion model. ASCs and BMCs were isolated from the same Lewis donor rats and grown to passage 2 (P2). Single-level bilateral posterolateral intertransverse process lumbar fusion surgery was performed on syngeneic rats divided into three experimental groups: clinical-grade bone graft substitute alone (CBGS); CBGS+ rat ASCs (rASC); and, CBGS+ rat BMCs (rBMC). Eight weeks postoperatively, fusion was evaluated via micro-CT, manual palpation and histology. In vitro analysis of the osteogenic capacity of rBMCs and rASCs was also performed. Results indicated that the average fusion volume in the rASC group was the largest and was significantly larger than the CBGS group. Although the rASC group displayed the highest fusion rates via micro-CT and manual palpation, this difference was not statistically significant. Cell-seeded grafts showed more histological bone formation than cell-free grafts. P2 rASCs and rBMCs displayed similar in vitro osteogenic differentiation capacities. Overall, this study showed that, when combined with a clinical-grade bone graft substitute in a rat model, rASCs cells yielded the largest fusion masses and comparable fusion results to rBMCs. These results add to growing evidence that ASCs provide an attractive alternative to BMCs for spinal fusion procedures.

The efficacy of a nanosynthetic bone graft substitute as a bone graft extender in rabbit posterolateral fusion

BACKGROUND CONTEXTSynthetic bone graft substitutes are commonly used in spinal fusion surgery. Preclinical data in a model of spinal fusion to support their efficacy is an important component in clinical adoption to understand how these materials provide a biological and mechanical role in spinal fusion. PURPOSETo evaluate the in vivo response of a nanosynthetic silicated calcium phosphate putty (OstP) combined with autograft compared to autograft alone or a collagen-biphasic calcium phosphate putty (MasP) combined with autograft in a rabbit spinal fusion model. STUDY DESIGNEfficacy of a nanosynthetic silicated calcium phosphate putty as an extender to autograft was studied in an experimental animal model of posterolateral spinal fusion at 6, 9, 12 and 26 weeks, compared to a predicate device. METHODSSkeletally mature female New Zealand White rabbits (70) underwent single level bilateral posterolateral intertransverse process lumbar fusion, using either autograft alone (AG), a nanosynthetic silicated calcium phosphate putty (OstP) combined with autograft (1:1), or a collagen-biphasic calcium phosphate putty (MasP) combined with autograft (1:1). Iliac crest autograft was harvested for each group, and a total of 2 cc of graft material was implanted in the posterolateral gutters per side. Fusion success was assessed at all time points by manual palpation, radiographic assessment, micro-CT and at 12 weeks only using non-destructive range of motion testing. Tissue response, bone formation and graft resorption were assessed by decalcified paraffin histology and by histomorphometry of PMMA embedded sections. RESULTSAssessment of fusion by manual palpation at the 12 week endpoint showed 7 out of 8 (87.5%) bilateral fusions in the OstP extender group, 4 out of 8 (50%) fusions in the MasP extender group, and 6 out of 8 (75%) fusions in the autograft alone group. Similar trends were observed with fusion scores of radiographic and micro-CT data. Histology showed a normal healing response in all groups, and increased bone formation in the OstP extender group at all timepoints compared to the MasP extender group. New bone formed directly on the OstP granule surface within the fusion mass while this was not a feature of the Collagen-Biphasic CaP material. After 26 weeks the OstP extender group exhibited 100% fusions (5 out of 5) by all measures, whereas the MasP extender group resulted in bilateral fusions in 3 out of 5 (60%), assessed by manual palpation, and fusion of only 20 and 0% by radiograph and micro-CT scoring, respectively. Histology at 26 weeks showed consistent bridging of bone between the transverse processes in the Ost P extender group, but this was not observed in the MasP extender group. CONCLUSIONSThe nanosynthetic bone graft substituted studied here, used as an extender to autograft, showed a progression to fusion between 6 and 12 weeks that was similar to that observed with autograft alone, and showed excellent fusion outcomes, bone formation and graft resorption at 26 weeks. CLINICAL SIGNIFICANCEThis preclinical study showed that the novel nanosynthetic silicated CaP putty, when combined with autograft, achieved equivalent fusion outcomes to autograft. The development of synthetic bone grafts that demonstrate efficacy in such models can eliminate the need for excessive autograft harvest and results from this preclinical study supports their effective use in spinal fusion surgery.

Comparing the efficacy of syngeneic iliac and femoral allografts with iliac crest autograft in a rat model of lumbar spinal fusion

BackgroundDespite widespread use of femoral-sourced allografts in clinical spinal fusion procedures and the increasing interest in using femoral reamer–irrigator–aspirator (RIA) autograft in clinical bone grafting, few studies have examined the efficacy of femoral grafts compared to iliac crest grafts in spinal fusion. The objective of this study was to directly compare the use of autologous iliac crest with syngeneic femoral and iliac allograft bone in the rat model of lumbar spinal fusion.MethodsSingle-level bilateral posterolateral intertransverse process lumbar spinal fusion surgery was performed on Lewis rats divided into three experimental groups: iliac crest autograft, syngeneic iliac crest allograft, and syngeneic femoral allograft bone. Eight weeks postoperatively, fusion was evaluated via microCT analysis, manual palpation, and histology. In vitro analysis of the colony-forming and osteogenic capacity of bone marrow cells derived from rat femurs and hips was also performed to determine whether there was a correlation with the fusion efficacy of these graft sources.ResultsAlthough no differences were observed between groups in CT fusion mass volumes, iliac allografts displayed an increased number of radiographically fused fusion masses and a higher rate of bilateral fusion via manual palpation. Histologically, hip-derived grafts showed better integration with host bone than femur derived ones, likely associated with the higher concentration of osteogenic progenitor cells observed in hip-derived bone marrow.ConclusionsThis study demonstrates the feasibility of using syngeneic allograft bone in place of autograft bone within inbred rat fusion models and highlights the need for further study of femoral-derived grafts in fusion.

235. Evaluation of a nanosynthetic silicated calcium phosphate putty in a posterolateral rabbit spinal fusion model

BACKGROUND CONTEXT Providing surgeons with an “off the shelf” bone graft substitute to aid in bone healing is advantageous providing the materials participate in and facilitate the desired goal of fusion. Synthetic calcium phosphate-based (CaP) materials are excellent candidates considering their comparable composition to bone mineral however other factors have been shown to play a role in osteogenic behavior of these materials including surface chemistry and surface topography. The preclinical rabbit posterolateral fusion developed by Professor Boden provides a robust means to evaluate the in vivo performance of graft materials in this challenging indication. PURPOSE This study aims to evaluate the in-vivo response of a nanosynthetic silicated CaP combined with autograft compared to autograft alone or a collagen-biphasic CaP combined with autograft in a rabbit spinal fusion model. STUDY DESIGN/SETTING Preclinical animal model. PATIENT SAMPLE N/A OUTCOME MEASURES N/A METHODS Following ethical approval, 70 skeletally mature female NZ white rabbits were divided into three groups: Group 1: Autograft alone (positive control); Group 2: Nanosynthetic Silicated CaP Putty + Autograft (50/50); Group 3: Collagen-Biphasic CaP Putty + Autograft (50/50). A single level posterolateral intertransverse process lumbar fusion was performed bilaterally after decortication of the transverse processes with a high-speed burr. Iliac crest autograft was harvested bilaterally in the positive control group and 2cc placed in the posterolateral gutters. The synthetic materials (1cc) were combined with autograft (1cc) and implanted. Fusion assessment at day 0 (n=2), 6, 9 weeks (n=5), 12 weeks (n=8) and 26 weeks (n=5) included harvest, manual palpation, radiographic assessment, micro-CT, non-destructive range of motion testing (12 weeks only) and decalcified paraffin and pmMA histology; no positive control group (autograft alone) was included at the 26-week time point. RESULTS The positive control group, autograft alone, performed consistent with this model in all endpoints. In the synthetic groups combined with autograft (Groups 2 and 3), radiographic incorporation was observed at the host transverse processes at 6 weeks and further into the fusion with time. Graft resorption and new bone formation was noted in the radiographs as well as the micro-computed tomography sections. Manual palpation was 7/8 at 12 weeks in the Nanosynthetic Silicated CaP Putty + Autograft group compared to 4/8 for the Collagen-Biphasic CaP Putty + Autograft. Histology showed normal healing and increased bone remodeling/formation with Nanosynthetic Silicated CaP Putty + Autograft at all timepoints compared to the Collagen-Biphasic CaP Putty + Autograft. New bone formed directly on the Nanosynthetic Silicated CaP material surface within the fusion mass while this was not a feature of the Collagen-Biphasic CaP material. By 26 weeks histology revealed consistent bridging of bone between the transverse processes in the Nanosynthetic Silicated CaP Putty + Autograft group, but this was not observed in the Collagen-Biphasic CaP group. CONCLUSIONS Developing nanosynthetic silicated CaP bone grafts that aim to mimic the size, structure and chemistry of bone mineral can eliminate the need for excessive autograft harvest and results from this preclinical study supports their effective use in spinal fusion. FDA DEVICE/DRUG STATUS Osteo 3 (the Nanosynthetic Silicated CaP Putty) (Investigational/Not approved)

Friday, September 28, 2018 10:30 AM–12:00 PM abstracts: deformity: technical factors: 165. Sagittal imbalance in degenerative scoliosis can be corrected by addressing only the symptomatic levels in carefully selected patients: a retrospective radiographic study

BACKGROUND CONTEXT The optimal method of addressing the positive sagittal imbalance often present in degenerative scoliosis (DS) is controversial. An important distinction is whether or not the etiology of the loss of lumbar lordosis (LL) and positive sagittal imbalance often seen on preoperative radiographs, is truly structural, or whether it is more positional in nature given that spinal stenosis is almost always present in DS patients who require surgery. This distinction directly affects the decision about whether or not, in addition to decompression, the entire curve needs to be addressed (regional or global fusion), or if a lesser local fusion of only the symptomatic levels can be performed with a satisfactory radiographic result. PURPOSE This study aims to determine (1) if the loss of LL often associated with DS is structural or rather largely due to positional factors secondary to spinal stenosis, and (2) if the entire DS curve or a lesser local fusion of only the neurologically symptomatic levels need to be addressed. STUDY DESIGN/SETTING Retrospective case series. PATIENT SAMPLE We obtained radiographic and clinical data on 114 consecutive patients. We included patients if they had loss of normal LL (positive sagittal imbalance) observed on preoperative radiographs and underwent a decompression and instrumented posterolateral inter-transverse process arthrodesis of only their neurologically symptomatic levels in the setting of DS. We excluded patients if they had undergone a previous fusion, concomitant osteotomy procedure, or any type of interbody arthrodesis, or had less than 6 months follow-up. Of the original 114 patients identified, 68 fulfilled the above criteria. OUTCOME MEASURES Pre- and postoperative radiographs were reviewed to measure lumbar lordosis, pelvic incidence, pelvic tilt, and sacral slope. Postoperative radiographs were also measured against preoperative extension lumbar radiographs and lumbar magnetic resonance imaging (MRI) to compare changes in radiographic pelvic parameters. METHODS Three independent members of the research team performed all measurements. Inter-rater reliability was calculated using a Pearson coefficient test and a strong correlation was considered if R>0.8. The three sets of measurements were then averaged for final statistical analysis. Relationships were considered significant at p RESULTS Average follow-up was 22 months. In addition to decompression, instrumented fusion was most commonly performed from L4 to S1 (N=17), followed by L3-S1 (N=12). The average PI found preop and postop was 52° and 53°, respectively. The average PT found preop and postop was 24° and 27°, respectively. Preoperative LL had a mean Cobb angle of 32° (range 2°–60°). Preop MRIs and lumbar extension radiographs revealed an average LL of 42° (range 10°–66°) and 48° (range 20°–74°), respectively. Initial postop radiographs (average 3 weeks) revealed that LL was corrected to a mean of 44° (range 14°–68°, p CONCLUSIONS We were able to correct LL to within 10° of the PI in this cohort of symptomatic DS patients with decompression and local instrumented fusion of symptomatic levels only. Loss of LL in symptomatic DS patients has a large positional component likely due to stenosis. Preoperative MRI and extension radiographs can be used to predict achievable postoperative LL correction.

Wednesday, September 26, 2018 7:35 AM–9:00 AM ePosters

BACKGROUND CONTEXT Posterolateral lumbar arthrodesis by autologous bone graft is a commonly performed procedure for the treatment of degenerative spine conditions. However, because autograft is associated with considerable disadvantages (eg, donor-site morbidity), alternative technologies are under investigation. Calcium phosphates (CaP) with submicron-scale surface topography have attracted attention due to their excellent performance in regeneration of bone defects. PURPOSE Evaluating the efficacy of a biphasic calcium phosphate (BCP) ceramic with a novel surface topography of submicron-scale epitaxial polygon crystals as autograft extender in the “Boden” rabbit posterolateral spine fusion model. STUDY DESIGN/SETTING BCP granules with the submicron-scale surface topography, either alone or combined with a fast-resorbing polymer binder, were mixed with autologous bone at a volume ratio of 50%:50%. Subsequently, grafts were implanted in the rabbit posterolateral spine environment with autologous bone graft as reference treatment, after which outcomes were assessed after 6, 9 and 12 weeks. PATIENT SAMPLE N/A. OUTCOME MEASURES Spinal fusion was determined by manual palpation (MP), X-ray and micro CT scan (based on the Lenke scale), mechanical testing (range of motion testing at 12 weeks only) and histology (decalcified and undecalcified). METHODS Sixty skeletally mature New Zealand white rabbits were divided into three groups and underwent single-level bilateral posterolateral intertransverse process spine arthrodesis at L4-5. Animals were euthanized 0, 6, 9 and 12 weeks for analysis of outcomes. RESULTS Based on all endpoints, the three groups showed an equivalent and gradual progression in bone formation and implant resorption during time leading to mature fusion masses at 12 weeks. No adverse reactions were observed in any of the groups. Bilateral fusion rates by MP were consistent with the literature (0%–20% at 6 weeks, 60% at 9 weeks, 75% at 12 weeks) and similar trends were seen by radiographic assessment (both X-ray and uCT). Flexion–extension and lateral bending results at 12 weeks demonstrated fusion of the treated level consistent with the MP results. By histology, from 6 weeks onwards, bone formation was observed on and between the BCP granules and within the residual autograft particles. At 9 and 12 weeks, progressive bone formation with developing marrow spaces was observed in all groups. Gradual resorption of BCP granules was evident with resorbing multinucleated or osteoclastic cells on the material surface and a decrease in material proportion over time. Histological fusion, ie, continuous bone formation between the transverse processes, for the three groups ranged 0%–20% at 6 weeks to 20%–40% at 9 weeks and 75%–87.5% at 12 weeks. CONCLUSIONS These results show that BCP with submicron-scale surface topography could achieve equivalent fusion rates to ABG when used as graft extender in a single level rabbit intertransverse process posterolateral lumbar arthrodesis model. During time, a progression in bone healing, graft resorption and remodeling was seen with all experimental endpoints, with consistency between the groups.

Efficacy comparison of Accell Evo3 and Grafton demineralized bone matrix putties against autologous bone in a rat posterolateral spine fusion model

Background ContextSpinal fusion procedures are intended to stabilize the spinal column for a multitude of disorders including abnormal curvature, traumatic instability, degenerative instability, and damage from infections or tumors. As an aid in the bone healing response, bone graft materials are used to bridge joints for arthrodesis and promote unions in pseudoarthrosis. Currently, the gold standard for stabilizing fusion masses in spinal procedures involves using the osteogenic, osteoinductive, and osteoconductive properties of autologous iliac crest corticocancellous bone. However, considerable morbidity is associated with harvesting the autologous graft. Donor site complications including infection, large hematomas, and pain have been reported at rates as high as 50% (Boden and Jeffrey, 1995). Biologically, the rate of bone repair dictates the rate at which the fusion mass will unite under autologous graft conditions. PurposeThe purpose of this study is to compare the quality and rate of fusion between Accell Evo3 and Grafton demineralized bone matrix (DBM), with the gold standard iliac crest bone graft (ICBG) as the control, in athymic rat posterolateral fusion. Study DesignThis study was a randomized, controlled study in a laboratory setting at the Hospital for Special Surgery in New York City. Blinded observations were made, which created an assessment of outcomes for successful fusions between each method. Patient SampleForty-eight (48) athymic rats were used in this study and underwent posterolateral lumbar fusion. They were assessed at either 3 weeks or 9 weeks to see the rate and efficacy of fusion. Outcome MeasuresOutcome measures will be the efficacy of the different bone grafts and their success rates of fusion in the rats. MethodsA comparison of the quality and rate of fusion between Accell Evo3® (DBM A) and Grafton (DBM B), with the gold standard iliac crest bone graft (ICBG) as the control, was performed using the established posterolateral intertransverse process on an athymic rat model. Materials were evaluated for fusion by several criteria, including manual palpation, standard and high-resolution radiographic imaging, micro-computed topography, and histologic analysis. Forty-eight (48) athymic rats received a bilateral intertransverse process fusion, using either bone from the iliac crest (control group), Accell Evo3, or Grafton. Twelve (12) rats (four from each group) were sacrificed at 3 weeks postoperatively, whereas the remaining thirty-six (36) were sacrificed at 9 weeks postoperatively. Three blinded observers examined the spines after the rats were euthanized, and they blindly assessed each rat for fusion success. ResultsManual palpation of the three different groups at 3 weeks postoperatively found successful fusion in 1 of 4 (25%) of the autologous bone graft (ABG) group and 4 of 4 (100%) of both DBM A and B groups. Manual palpation of the remaining animals that were sacrificed at 9 weeks postoperatively showed successful fusion in 4 of 12 (33%) of the ABG group, 8 of 12 (66%) of the DBM A group, and 12 of 12 (100%) of the DBM B group. Radiography found that 9 of 16 (56%) of the ABG group and 16 of 16 (100%) of both DBM Putty A and B groups had fused. Histologic analysis of the ABG group demonstrated less mature and less organized osteoid at both 3 and 9 weeks than the DBM Putty A and B groups. Nondestructive mechanical testing demonstrated increased stiffness in 4-point bending of both DBM A and B compared with ABG. ConclusionsBoth DBM-treated groups achieved a significantly higher rate of fusion than the ABG-treated group at 9 weeks in this model. Successful fusion was also demonstrated in the DBM-treated groups at 3 weeks.

A Quantitative Study of the Effects of High-Dose Parathyroid Hormone (1–34) and Low-Dose rhBMP-2 on Posterolateral Lumbar Spine Fusion in a Rabbit Model

Introduction Parathyroid hormone (1–34) (PTH), which is commonly used in the treatment of osteoporosis, has been shown to increase bone mass and reduce the risk of fracture. Previously, we demonstrated that, while intermittent low-dose (10 µg/kg) PTH (1–34) treatment resulted in increased fusion mass volume, it did not improve biomechanical stiffness over the use of autograft alone, and did not improve fusion when combined with BMP-2 treatment (Lina et al, Spine [Phila Pa 1976]. 2014; 39(5):347). However, studies in other rodent models have indicated that the effects of PTH (1–34) on spinal fusion may be dose-dependent (Ming et al, Spine [Phila. Pa. 1976]. 2012; 37(15): 1275; Abe et al, Bone. 2007; 41(5): 775). These rat and mouse studies used PTH (1–34) doses as high as 20 to 40 µg/kg compared with the 10 µg/kg used in our experiments. We thus aim to evaluate whether increasing the dose of PTH (1–34) administered increases both the volume and biomechanical stiffness of the resulting fusion masses and/or exhibits synergistic effects with low-dose BMP-2 treatment. Materials and Methods A total of 60 rabbits underwent posterolateral intertransverse process lumbar spinal fusion surgery, and were assigned to one of the six groups depending upon the fusion grafting group employed: iliac crest autograft alone ( n = 10), iliac crest autograft + 20 µg/kg PTH (1–34) ( n = 10), iliac crest autograft + 40 µg/kg PTH (1–34) ( n = 10), rhBMP-2 alone ( n = 10), rhBMP-2 + 20 µg/kg PTH (1–34) and, rhBMP-2 + 40 µg/kg PTH (1–34). Fusion mass and quality were assessed using manual palpation, CT analysis of fusion mass volume, and four-point nondestructive biomechanical testing. Results/Conclusions: Although manual palpation is used as the standard measure of fusion, we found that CT analysis proved more effective in differentiating fused masses based on bone formation, not fibrous tissue. Both the PTH (1–34) group and BMP-2 increased bone density and volume of the fusion masses. Biomechanical analysis of fusion mass strength is currently underway.

Accelerated postero-lateral spinal fusion by collagen scaffolds modified with engineered collagen-binding human bone morphogenetic protein-2 in rats.

Bone morphogenetic protein-2 (BMP-2) is a potent osteoinductive cytokine that plays a critical role in bone regeneration and repair. However, its distribution and side effects are major barriers to its success as therapeutic treatment. The improvement of therapy using collagen delivery matrices has been reported. To investigate a delivery system on postero-lateral spinal fusion, both engineered human BMP-2 with a collagen binding domain (CBD-BMP-2) and collagen scaffolds were developed and their combination was implanted into Sprague-Dawley (SD) rats to study Lumbar 4–5 (L4–L5) posterolateral spine fusion. We divided SD rats into three groups, the sham group (G1, n = 20), the collagen scaffold-treated group (G2, n = 20) and the BMP-2-loaded collagen scaffolds group (G3, n = 20). 16 weeks after surgery, the spines of the rats were evaluated by X-radiographs, high-resolution micro-computed tomography (micro-CT), manual palpation and hematoxylin and eosin (H&E) staining. The results showed that spine L4–L5 fusions occurred in G2(40%) and G3(100%) group, while results from the sham group were inconsistent. Moreover, G3 had better results than G2, including higher fusion efficiency (X score, G2 = 2.4±0.163, G3 = 3.0±0, p<0.05), higher bone mineral density (BMD, G2: 0.3337±0.0025g/cm3, G3: 0.4353±0.0234g/cm3. p<0.05) and more bone trabecular formation. The results demonstrated that with site-specific collagen binding domain, a dose of BMP-2 as low as 0.02mg CBD-BMP-2/cm3 collagen scaffold could enhance the posterolateral intertransverse process fusion in rats. It suggested that combination delivery could be an alternative in spine fusion with dramatically decreased side effects caused by high dose of BMP-2.

Early-term and Mid-term Histologic Events During Single-level Posterolateral Intertransverse Process Fusion With rhBMP-2/Collagen Carrier and a Ceramic Bulking Agent in a Nonhuman Primate Model

We used a nonhuman primate lumbar intertransverse process arthrodesis model to evaluate biological cascade of bone formation using different carrier preparation methods with a single dose of recombinant human bone morphogenetic protein-2 (rhBMP-2) at early time points. To examine early-term/mid-term descriptive histologic and computerized tomographic events in single-level uninstrumented posterolateral nonhuman primate spinal fusions using rhBMP-2/absorbable collagen sponge (ACS) combined with ceramic bulking agents in 3 different configurations. rhBMP-2 on an ACS carrier alone leads to consistent posterolateral lumbar spine fusions in lower-order animals; however, these results have been difficult to replicate in nonhuman primates. Twelve skeletally mature, rhesus macaque monkeys underwent single-level posterolateral arthrodesis at L4-L5. A hydroxyapatite/β-tricalcium phosphate ceramic bulking agent in 3 formulations was used in the treatment groups (n=3). When used, rhBMP-2/ACS at 1.5 mg/cm (3.0 mg rhBMP-2) was combined with 2.5 cm of ceramic bulking agent per side. Animals were euthanized at 4 and 12 weeks postoperative. Computerized tomography scans were performed immediately postoperatively and every 4 weeks until they were euthanized. Sagittal histologic sections were evaluated for bone histogenesis and location, cellular infiltration of the graft/substitute, and bone remodeling activity. Significant histologic differences in the developing fusion appeared between the 3 rhBMP-2/ACS treatment groups at 4 and 12 weeks. At 4 weeks, bone formation appeared to originate at the transverse process and the intertransverse membrane. Cellular infiltration was greatest in granular ceramic groups compared with matrix ceramic group. Minimal to no residual ACS was identified at the early time point. At 12 weeks, marked ceramic remodeling was observed with continued bone formation noted in all carrier groups. At the early time period, histology showed that bone formation appeared to originate at the transverse processes and the intertransverse membrane, indicating that the dorsal muscle bed may not be the only location for bone formation. Histology also showed that the collagen carrier for rhBMP-2 is mostly resorbed by 4 weeks. Our results and previous literature indicate that ceramic bulking agents are needed to provide resistance to compression caused by paraspinal muscles on the fusion bed in the posterolateral environment. Histology showed that ceramic bulking agents may offer long-term scaffolding and a structure to supporting bone formation of the developing fusion mass.

Quantitative Study of Parathyroid Hormone (1-34) and Bone Morphogenetic Protein-2 on Spinal Fusion Outcomes in a Rabbit Model of Lumbar Dorsolateral Intertransverse Process Arthrodesis

A posterolateral rabbit spinal fusion model was used to evaluate the effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) and teriparatide (PTH [1-34]) used individually and in combination on spinal fusion outcomes. To test the efficacy of parathyroid hormone on improving spinal fusion outcomes when used with BMP-2. Of the more than 250,000 spinal fusion surgical procedures performed each year, 5% to 35% of these will result in pseudarthrosis. Growing controversy on the efficacy and cost of rhBMP-2 for improving spinal fusion outcomes has presented a challenge for clinicians. Research into PTH as an adjunct therapy to rhBMP-2 for spinal fusion has not yet been investigated. Forty-eight male New Zealand white rabbits underwent bilateral posterolateral intertransverse process arthrodesis surgery at the L5-L6 level. Animals were divided into 6 groups. Two groups were treated with autograft alone or autograft and PTH (1-34), whereas the other 4 groups were treated with low-dose rhBMP-2 alone, high-dose rhBMP-2 alone, or either dose combined with PTH (1-34). All animals were euthanized 6 weeks after surgery. The L4-L7 spinal segment was removed and assessed using manual palpation, computed tomography (CT), and biomechanical testing. CT assessments revealed fusion in 50% of autograft controls, 75% of autograft PTH (1-34) animals, 87.5% in the 2 groups treated with low-dose rhBMP-2, and 100% in the 2 groups treated with high-dose rhBMP-2. CT volumetric analysis demonstrated that all groups treated with biologics had fusion masses that were on average significantly larger than those observed in the control group (P < 0.0001). Biomechanical data demonstrated no statistical difference between controls, PTH (1-34), and low-dose rhBMP-2 in any testing orientation. PTH (1-34) did not increase bending stiffness when used adjunctively with either low-dose or high-dose rhBMP-2. Although intermittent teriparatide administration results in increased fusion mass volume, it does not improve biomechnical stiffness over use of autograft alone. When delivered concurrently with high- and low-dose rhBMP-2, teriparatide provided no statistically significant improvement in biomechanical stiffness. N/A.

Posterolateral spinal fusion with ostegenesis induced BMSC seeded TCP/HA in a sheep model

Autogenous bone graft is the gold standard for fusion procedure. However, pain at donor site and inconsistent outcome have left a surgeon to venture into some other technique for spinal fusion. The objective of this study was to determine whether osteogenesis induced bone marrow stem cells with the combination of ceramics granules (HA or TCP/HA), and fibrin could serve as an alternative to generate spinal fusion. The sheep's bone marrow mesenchymal stem cells (BMSCs) were aspirated form iliac crest and cultured for several passages until confluence. BMSCs were trypsinized and seeded on hydroxyapatite scaffold (HA) and tricalcium phosphate/hydroxyapatite (TCP/HA) for further osteogenic differentiation in the osteogenic medium one week before implantation. Six adult sheep underwent three-level, bilateral, posterolateral intertransverse process fusions at L1-L6. Three fusion sites in each animal were assigned to three treatments: (a) HA constructs group/L1-L2, (b) TCP/HA constructs group/L2-L3, and (c) autogenous bone graft group/L5-L6. The spinal fusion segments were evaluated using radiography, manual palpation, histological analysis and scanning electron microscopy (SEM) 12 weeks post implantation. The TCP/HA constructs achieved superior lumbar intertransverse fusion compared to HA construct but autogenous bone graft still produced the best fusion among all.

Effect of intermittent PTH (1–34) on posterolateral spinal fusion with iliac crest bone graft in an ovariectomized rat model

Intermittent treatment with high-dose parathyroid hormone (PTH) enhances the quantity and quality of the fusion callus and reduces healing time of posterolateral spinal fusion with autologous iliac bone grafts in ovariectomized osteoporotic female Sprague-Dawley rats. Intermittent PTH (1-34) could be an appropriate adjunctive therapy for osteoporotic patients undergoing posterolateral intertransverse process fusion. The study was designed to test the hypothesis that intermittent administration of PTH improves spinal fusion rates in a randomized controlled, ovariectomized osteoporotic rat spinal fusion model. Thirty-six 10-week-old Sprague-Dawley rats were ovariectomized and underwent bilateral posterolateral L4-L5 spinal fusion with autologous iliac bone graft 6 weeks later. The experimental (PTH) group (18 rats) received daily subcutaneously administered injections of PTH (1-34) at 30 μg/kg/day starting on the day of operation. The control group (18 rats) received a subcutaneously administered injection of normal saline of the same volume. Nine rats from each group were sacrificed at 4 and 6 weeks. After sacrifice, the L4-L5 vertebral segments were removed and analyzed by plain radiographs, μ-CT, histomorphometry, and serum bone metabolism marker. The PTH group had a significantly higher fusion rate and X-ray fusion score than the control group at 4 and 6 weeks (p < 0.05). μ-CT and histological analysis showed that the fusion bone volume and cortical thickness for the PTH group were significantly higher than those for the control group at 4 and 6 weeks (p < 0.05). Metabolic marker analysis also showed significant difference between the two groups. The serum osteocalcin was significantly higher in the PTH group at 4 and 6 weeks, and levels of N-terminal peptide of type I collagen were significantly higher at 4 weeks (p < 0.05). Intermittent treatment with high-dose PTH enhances the quantity of the fusion callus and reduces the healing time of posterolateral spinal fusion with autologous iliac bone grafts in ovariectomized osteoporotic female Sprague-Dawley rats.

Bone Marrow Enhances the Performance of rhBMP-2 in Spinal Fusion

Reliable and rapid bone formation is the goal of biologics and cell-based spinal fusion technologies. While no cell-based therapy alone has been successful, recombinant human bone morphogenetic protein-2 (rhBMP-2) has been successfully used in a wide spectrum of patients undergoing a variety of spinal fusion procedures since its approval by the United States Food and Drug Administration (FDA) in 2002. However, the question remains how to improve the biologic efficiency, or osteoinductivity, of rhBMP-2 for successful application in the most challenging patients undergoing spinal fusion or to reduce the doses currently required. The present study investigated how varying the cellular environments through the addition of freshly harvested bone marrow aspirate (BMA) modulates rhBMP-2 efficiency. An L4-L5 posterolateral intertransverse process spinal fusion procedure was performed in Lewis rats. The implants were a subeffective concentration of 0.006 mg/mL of rhBMP-2/two absorbable collagen sponges (ACS) plus directly applied fresh syngeneic BMA transplants (n = 18), 0.006-mg/mL rhBMP-2/two ACS/side (n = 12), 0.006-mg/mL rhBMP-2/one ACS/side (n = 12), or BMA/one ACS/side (n = 6). Rats were killed at eight weeks and were evaluated with use of manual palpation, radiographs, and biomechanical testing. BMA plus 0.006-mg/mL rhBMP-2/ACS significantly increased the L4-L5 fusion rate to 89% (sixteen of eighteen) compared with a base fusion rate of 33% (four of twelve) to 50% (six of twelve) for rats implanted with rhBMP-2/ACS (p < 0.05), with no difference in strength or stiffness between conditions. No fusion or bone formation was observed in the six rats that received BMA/ACS alone. Less rhBMP-2 was needed for effect when mixed with BMA. A nearly twofold increase in the fusion rate was found when BMA was mixed with a deliberate subeffective concentration of rhBMP-2. There was no improvement in terms of fusion strength or stiffness.

Influence of blood supply of paraspinal muscle on a novel posterior lumbar fusion

Objective To construct a novel posterior lumbar spine fusion canine model,evaluate the result of bone graft healing and compare with the conventional posterolateral lumbar intertransverse process fusion.Methods Thirty-two mongrel dogs were randomly assigned to experimental group or control group.The animals were sacrificed at 8th week or 16th week after surgery.The lumbar spines were evaluated radiologically and histologically,and the serum biochemical indicators of bone were examined.Results The radiographic and histological analysis at 8 and At 16th week after the surgery demonstrated that the score of fusion in the treatment group was statistically significantly higher than in the control group (P ＜0.05).At 8th week,the radiographic fusion rate was 57.1％ and 21.4％ in the treatment and control groups,respectively; and at 16th week,the fusion rate was 93.8％ and 62.5％,respectively.The levels of bone alkaline phosphatase(BALP),osteocalcin(OC) and type Ⅰ collagen telopeptide(CTX-1) in the treatment group were significantly higher than in the control group.The dogs in the treatment group all the time were at higher state of bone turnover.Conclusion The posterior lumbar fusion is a novel and useful method that makes the bone graft position more precise and stable and increases the rate of fusion.This method can decreases the incidence of pseudarthrosis. Key words: Lumbar fusion; Paraspinal muscle; Muscle bone flaps; Blood supply

Novel oxysterols have pro-osteogenic and anti-adipogenic effects in vitro and induce spinal fusion in vivo

Stimulation of bone formation by osteoinductive materials is of great clinical importance in spinal fusion surgery, repair of bone fractures, and in the treatment of osteoporosis. We previously reported that specific naturally occurring oxysterols including 20(S)-hydroxycholesterol (20S) induce the osteogenic differentiation of pluripotent mesenchymal cells, while inhibiting their adipogenic differentiation. Here we report the characterization of two structural analogues of 20S, Oxy34 and Oxy49, which induce the osteogenic and inhibit the adipogenic differentiation of bone marrow stromal cells (MSC) through activation of Hedgehog (Hh) signaling. Treatment of M2-10B4 MSC with Oxy34 or Oxy49 induced the expression of osteogenic differentiation markers Runx2, Osterix (Osx), alkaline phosphatase (ALP), bone sialoprotein (BSP), and osteocalcin (OCN), as well as ALP enzymatic activity and robust mineralization. Treatment with oxysterols together with PPARγ activator, troglitazone (Tro), inhibited mRNA expression for adipogenic genes PPARγ, LPL, and aP2, and inhibited the formation of adipocytes. Efficacy of Oxy34 and Oxy49 in stimulating bone formation in vivo was assessed using the posterolateral intertransverse process rat spinal fusion model. Rats receiving collagen implants with Oxy 34 or Oxy49 showed comparable osteogenic efficacy to BMP2/collagen implants as measured by radiography, MicroCT, and manual inspection. Histological analysis showed trabecular and cortical bone formation by oxysterols and rhBMP2 within the fusion mass, with robust adipogenesis in BMP2-induced bone and significantly less adipocytes in oxysterol-induced bone. These data suggest that Oxy34 and Oxy49 are effective novel osteoinductive molecules and may be suitable candidates for further development and use in orthopedic indications requiring local bone formation.

The Influence of Orthotopic Paraspinal Muscle-Pediculated Bone Flaps on Posterior Spinal Fusion in a Canine Model

An experimental animal study with randomized, control design was conducted using a dog model. OBJECTIVE.: To construct a novel posterior lumbar spine fusion model with orthotopic paraspinal muscle-pediculated bone flaps and to compare with the conventional posterolateral lumbar intertransverse process fusion. Previous studies have demonstrated that paraspinal musculature provided important vascular ingrowth into the fusion mass. However, the blood supply of paraspinal muscles was still not sufficiently used in spinal fusion. In this study, we assessed the significant role of orthotopic paraspinal muscle-pediculated bone flaps in the healing of bone graft in spinal fusion. Thirty-two mongrel dogs were randomly assigned to 2 groups to undergo either posterior spinal fusion with orthotopic paraspinal muscle-pediculated bone flaps (treatment group) or posterolateral intertransverse process fusion (control group) at L5-L6 segment. All the fusions used an autologous bone graft obtained from bilateral posterior iliac crests. The animals were killed at 8 weeks or 16 weeks after surgery. The lumbar spines were evaluated by radiology, histology, and biomechanics. Treatment groups showed an increasing trend on radiographic grades, manual palpation, and biomechanical stiffness compared with control groups at 8 and 16 weeks. Histologic analysis revealed that there was more mature woven bone in the treatment group than the controls at both points. At 8 weeks, the histologic fusion rate was 21.4% in the treatment group, 50% in the control group (P < 0.05); and at 16 weeks the fusion rate was 50% and 87.5% in the control group and the treatment group, respectively (P < 0.05). The use of orthotopic paraspinal musclepediculated bone flaps increased the rate and quality of fusion in the experimental spinal fusion model. This suggested that the novel posterior spinal fusion could be further developed for clinical application.