Intertransverse Process Fusion Research Articles

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.

Tissue-engineered bone used in a rabbit model of lumbar intertransverse process fusion: A comparison of osteogenic capacity between two different stem cells.

Spinal fusion serves an important role in the reconstruction of spinal stability via restoration of the normal spinal sequence and relief of pain. Studies have demonstrated that the fusion rate is mainly associated with the osteogenic capacity of the implanted graft. Mesenchymal stem cells (MSCs) have been successfully isolated from human degenerated cartilage endplate (CEP) and designated as CEP-derived stem cells (CESCs). Previous studies have suggested that CESCs possesses in vitro and in vivo chondrogenic potential superior to that of bone marrow (BM)-MSCs. In addition, CESCs have shown a stronger in vitro osteogenic ability. The present study aimed to further determine the in vivo three-dimensional osteogenesis efficacy of CESCs for spinal fusion. Tissue-engineered bone grafts were transplanted into a rabbit model of posterolateral lumbar intertransverse process fusion using CESCs and BM-MSCs as seed cells composited with porous hydroxyapatite (PHA). The results of manual palpation and computed tomography (CT) scan reconstruction indicated that the CESCs/PHA group had a higher fusion rate than the BM-MSCs/PHA group, although the difference was not observed to be statistically significant. In addition, RT-qPCR results revealed that the in vitro CESCs/PHA composite expressed significantly higher levels of osteogenic-specific mRNA compared with the BM-MSCs/PHA composite. Finally, micro-CT and semi-quantitative histological analysis further demonstrated that the newly formed bone quality of the CESCs/PHA group was significantly higher than that of the BM-MSCs/PHA group in the intertransverse process fusion model. Therefore, the study indicated that CESCs possess superior in vivo osteogenesis capacity compared with BM-MSCs, and might serve as an important alternative seed cell source for bone tissue engineering. These results may provide the foundation for a biological solution to spinal fusion or other bone defect issues.

Tissue Engineered Bone Differentiated From Human Adipose Derived Stem Cells Inhibit Posterolateral Fusion in an Athymic Rat Model.

Biological augmentation spinal arthrodesis trial in athymic rats. To assess the efficacy of tissue-engineered bone to promote L4-L5 intertransverse process fusion in an athymic rat model. Each year in the United States, over 400,000 spinal fusion surgeries are performed requiring bone graft. The current gold standard for posterolateral lumbar fusion is autogenous iliac crest bone graft (ICBG), but the harvesting of ICBG is associated with increased operative time and significant complications. This being the case, an alternative cost-effective bone graft source is needed. Bovine bone cores were sterilized and decellularized for scaffold production. Human adipose derived mesenchymal stem cells (ADSC) were obtained and verified by tridifferentiation testing and seeded onto dried scaffolds. The seeded cores were cultured for 5 weeks in culture medium designed to mimic endochondral ossification and produce hypertrophic chondrocytes. Single-level intertransverse process fusions were performed at the L4-L5 level of 31 athymic rats. Fifteen rats were implanted with the hypertrophic chondrocyte seeded scaffold and 16 had scaffold alone. Half of the study rats were sacrificed at 3 weeks and the other half at 6 weeks. Spinal fusion was assessed using 2D and 3D micro computed tomography (μCT) analysis and tissue histology. At 3 weeks, none of the tissue engineered rats had partial or complete fusion, whereas 62.5% of the decellularized rats fused and another 12.5% had partial fusions (P = 0.013). At 6 weeks, none of the tissue engineered rats fused and 50% had partial fusions, whereas 87.5% of the decellularized rats fused (P = 0.002). Tissue engineered bone composed of hypertrophic chondrocytes inhibits posterolateral fusion in an athymic rat model and therefore does not represent a promising cost-effective bone graft substitute. N/A.

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.

Histological Evaluation of Hydroxyapatite Granules with and without Platelet-Rich Plasma versus an Autologous Bone Graft: Comparative study of biomaterials used for spinal fusion in a New Zealand white rabbit model.

Hydroxyapatite (HA) has osteoconductive properties and is widely used as a bone graft substitute. Platelet-rich plasma (PRP) is an autologous product with osteoinductive effects. Hypothetically, a combination of both would augment the bone formation effect of HA and widen its application in spinal fusion surgeries. This study aimed to compare new bone formation with HA granules alone and in combination with PRP versus an autologous bone graft during a lumbar intertransverse process spinal fusion. A total of 16 adult New Zealand white rabbits underwent single-level bilateral intertransverse process fusion at the L5-L6 vertebrae. One side of the spine received either HA granules alone or a combination of HA granules and PRP, while the contralateral side received an autologous bone graft. Four animals each from the HA group and the HA plus PRP group versus the autograft group were assessed either at six or 16 weeks by undecalcified histology and histomorphometry. The mean percentage of new bone areas over the corresponding fusion masses were compared between groups. No significant difference in new bone formation was observed between the HA and HA plus PRP groups at six or 16 weeks. The autograft group had significantly more new bone formation at six and 16 weeks (P = 0.004 and <0.001, respectively). An autologous bone graft remains superior to HA granules, with or without PRP. HA granules demonstrated an excellent osteoconductive scaffold but had poor biodegradability. While PRP enhances the properties of HA granules, these biomaterials do not have a synergistic effect.

Good clinical outcomes and fusion rate of facet fusion with a percutaneous pedicle screw system for degenerative lumbar spondylolisthesis: minimally invasive evolution of posterolateral fusion.

A retrospective clinical and radiographical study. To assess the clinical outcomes and fusion rate of facet fusion (FF) for degenerative lumbar spondylolisthesis (DLS). On the basis of the long-term clinical and radiological follow-up studies of posterolateral fusion (PLF)-that is, intertransverse process fusion with pedicle screw instrumentation-for DLS, we recognized that FF alone would be sufficient for spinal fusion. Eighty-eight patients who underwent FF for single-level DLS were retrospectively reviewed after at least 1 year of follow-up. The control group comprised 21 patients who underwent conventional PLF. The operative technique involved a 5-cm midline skin incision, bilateral laminar fenestration, and FF with autologous bone harvested from the spinous process. Percutaneous pedicle screws were then inserted through the fascia. The fusion rate of FF was evaluated using computed tomography, and the change in the range of motion at the fused level was assessed on flexion-extension lateral radiographs. The Japanese Orthopaedic Association Back Pain Evaluation Questionnaire was used to assess the therapeutic effectiveness of FF. The results of the Roland-Morris Disability Questionnaire and the visual analogue scales of low back pain, buttock and lower limb pain, and buttock and lower limb numbness were evaluated. The fusion rate was 88.6% (78/88 cases). Among 10 patients with inadequate fusion, the average range of motion significantly decreased from 14.4° preoperatively to 4.3° postoperatively. The Japanese Orthopaedic Association Back Pain Evaluation Questionnaire category scores demonstrated therapeutic effectiveness in 93.0% of the patients for walking ability and in 73.0% of the patients for low back pain. The average preoperative scores of the Roland-Morris Disability Questionnaire and the visual analogue scales of low back pain, buttock and lower limb pain, and buttock and lower limb numbness were significantly reduced postoperatively in the FF group. FF achieved good clinical outcomes that were superior to those of conventional PLF with a comparable fusion rate. It is useful for managing DLS and is a minimally invasive evolution of PLF. 3.

Effect of serum nicotine level on posterior spinal fusion in an in vivo rabbit model

Background contextCigarette smoking has a deleterious effect on spinal fusion. Although some studies have implied that nicotine is primarily responsible for poor fusion outcomes, other studies suggest that nicotine may actually stimulate bone growth. Hence, there may be a dose-dependent effect of nicotine on posterior spinal fusion outcomes. PurposeThe purpose of this study was to determine if such a relationship could be shown in an in vivo rabbit model. Study design/SettingThis is a prospective in vivo animal study. MethodsTwenty-four adult male New Zealand white rabbits were randomly divided into four groups. All groups received a single-level posterolateral, intertransverse process fusion at L5–L6 with autologous iliac crest bone. One group served as controls and only underwent the spine fusion surgery. Three groups received 5.25-, 10.5-, and 21-mg nicotine patches, respectively, for 5 weeks. Serum nicotine levels were recorded for each group. All animals were euthanized 5 weeks postoperatively, and spinal fusions were evaluated radiographically, by manual palpation, and biomechanically. Statistical analysis evaluated the dose response effect of outcomes variables and nicotine dosage. This study was supported by a portion of a $100,000 grant from the Orthopaedic Research and Education Foundation. Author financial disclosures were completed in accordance with the journal's guidelines; there were no conflicts of interests disclosed that would have led to bias in this work. ResultsThe average serum levels of nicotine from the different patches were 7.8±1.9 ng/mL for the 5.25-mg patch group; 99.7±17.7 ng/mL for the 10.5-mg patch group; and 149.1±24.6 ng/mL for the 21-mg patch group. The doses positively correlated with serum concentrations of nicotine (correlation coefficient=0.8410, p<.001). The 5.25-mg group provided the best fusion rate, trabeculation, and stiffness. On the basis of the palpation tests, the fusion rates were control (50%), 5.25 mg (80%), 10.5 mg (50%), and 21 mg (42.8%). Radiographic assessment of trabeculation and bone incorporation and biomechanical analysis of bending stiffness ratio were also greatest in the 5.25-mg group. Radiographic evaluation showed a significant (p=.0446) quadratic effect of nicotine dose on spinal fusion. ConclusionsThe effects of nicotine on spinal fusion are complex, may be dose dependent, and may not always be detrimental. The uniformly negative effects of smoking reported in patients undergoing spinal fusion may possibly be attributed to the other components of cigarette smoke.

Mesenchymal stem cells expressing baculovirus-engineered BMP-2 and VEGF enhance posterolateral spine fusion in a rabbit model

Background contextMesenchymal stem cell (MSC)–based cell therapy and gene transfer have converged and show great potential for accelerating bone healing. Gene therapy can provide more sustained expression of osteogenic factors such as bone morphogenetic protein-2 (BMP-2). We previously demonstrated that low-dose BMP-2 enhanced spinal posterolateral fusion by MSCs in a rabbit model. Herein, we genetically modified rabbit MSCs with a recombinant baculovirus encoding BMP-2 (Bac-CB) and vascular endothelial growth factor (Bac-VEGF) seeded into porous scaffolds to enhance spinal fusion. PurposeThis study evaluates the success rate of the MSC-based cell therapy and gene transfer approach for single-level posterolateral spine fusion. We hypothesize that combining three-dimensional tricalcium phosphate (TCP) scaffolds and genetically modified allogeneic MSCs with baculovirus-mediated growth factor expression would increase the success rate of spinal fusion. Study designThe study design was based on an animal model (approved by the Institutional Animal Care and Use Committee) using 18 adult male New Zealand rabbits. MethodsThis study included 18 male New Zealand rabbits, weighing 3.5 to 4 kg. Allogeneic bone marrow–derived MSCs were isolated and genetically modified with Bac-CB and Bac-CV seeded onto TCP scaffolds (MSC/Bac/TCP). The animals were divided into three groups according to the material implanted into the bilateral L4–L5 intertransverse space: TCP scaffold (n=6), MSC/TCP (n=6), and MSC/Bac/TCP (n=6). After 12 weeks, the rabbits were euthanized for radiographic examination, manual palpation, and histologic study. ResultsBilateral fusion areas in each animal were evaluated independently. The radiographic fusion rates at 12 sites were 0 of 12 in the TCP scaffold group, 4 of 12 in the MSC/TCP group, and 10 of 12 in the MSC/Bac/TCP group. By manual palpation, there were zero solid fusions in the TCP scaffold group, two solid fusions in the MSC/TCP group, and five solid fusions in the MSC/Bac/TCP group. Fusion rates were significantly greater in the MSC/Bac/TCP group. ConclusionsThe results indicate the potential of using baculovirus as a vector for gene/cell therapy approaches to improve bone healing and support the feasibility of using allogeneic MSCs for inducing bone formation and intertransverse process fusion.

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.

Effect of Pentoxifylline on Spinal Fusion

Randomized, double-blinded, animal model. The objective of this study is to evaluate the effect of pentoxifylline (PTX) on spinal fusion in a rabbit model. Previous studies assert that PTX increases new bone formation. Because PTX seems to have these profound effects on bone metabolism, it may be hypothesized that it may enhance spinal fusion. Twenty-four New Zealand white rabbits were randomized and each received single-level posterolateral, inter-transverse process fusion with autologous iliac crest. In group 1, 12 male New Zealand white rabbits were treated with intravenous PTX treatment in 100-mg/kg/day dose after the surgical procedure. In group 2, 12 received no PTX medication and were accepted as the control group. Nine weeks after surgery, the animals were killed. The spines were tested via a manual palpation test, biomechanical testing, plain radiography, computed tomographic scans, and histomorphometric analysis. The fusion rates of manual palpation were 40% in the control group and 80% in the PTX group (P = 0.17). Using a 5-grade radiographical system, the mean fusion grade was 2.4 in the control group and 3.1 in PTX group (P = 0.012). Total displacement of the fused level for the control group under flexion and extension was 0.2515 mm and was lower for the PTX-treated group: 0.1266 mm (P = 0.012). In the control group, the mean bone volume of the fusion mass determined from computed tomographic analysis was 4.0678 cm, whereas in the PTX group it was 4.7802 cm (P = 0.009). The mean trabecular bone area was 14% and 19% for the control and PTX groups, respectively (P = 0.002). The differences between groups was statistically significant in terms of radiological fusion grading, biomechanical testing, volume of the fusion mass, and percentage of trabecular bone area. These results suggest that PTX may have a beneficial effect on spinal fusion. 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.

Dorsal root ganglion electrical stimulation promoted intertransverse process spinal fusion without decortications and bone grafting: a proof-of-concept study

Background contextPeriosteum, endosteum, and bone are innervated by sensory nerves expressing calcitonin gene-related peptide (CGRP), which is a known osteoanabolic peptide and plays an important role in fracture healing and spinal fusion. Synthesis and release of CGRP are found in sensory neurons located in the dorsal root ganglions (DRGs) and can be upregulated by electrical stimulation (ES) at DRG. PurposeTo prove our study hypothesis on the potential of precise ES at DRG through implantable microelectrical stimulation system (IMESS) for its effect on promoting spinal fusion in a rat model without decortications and bone grafting. Study designAn experimental animal study. MethodsA novel IMESS was developed for stimulating L4–L6 DRG in rats. Sixteen rats were used and divided equally into the control group without ES and the ES group, with a daily 20 minutes ES to DRG for 6 weeks. At the end of 6 weeks, radiography and microcomputed tomography were conducted to evaluate new bone formation and spinal fusion. Bilateral L4–L6 DRGs were harvested for immunohistochemistry and quantification of neurons with upregulated CGRP expression. ResultsIn the ES group, rate of radiographic fusion with complete and uninterrupted bony bridging was 100% (8/8) at the right L4/L5 transverse processes and 75% (6/8) at the right L5/L6 transverse processes. Bony callus formation was absent at the left L4–L6 transverse processes in the ES group and in bilateral L4–L6 transverse processes in the control group. ConclusionsWe proved for the first time that precise ES at DRG through IMESS effectively promoted intertransverse process fusion in rat model without decortications and bone grafting. Electrical stimulation at DRG might be an attractive minimal invasive bioengineering approach and an alternative therapy for intertransverse process fusion that is increasingly being used for the treatment of degenerative spine disorders.

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.

Negative Effect of Rapidly Resorbing Properties of Bioactive Glass-Ceramics as Bone Graft Substitute in a Rabbit Lumbar Fusion Model

BackgroundBioactive glass-ceramics have the ability to directly bind to bones and have been widely used as bone graft substitutes due to their high osteoconductivity and biocompatibility. CaO-SiO2-P2O5-B2O3 glass-ceramics are known to have good osteoconductivity and are used as bone graft extenders.MethodsThis study aimed to evaluate the effects of the resorbing properties of glass-ceramics in bone fusion after producing and analyzing three types of CaO-SiO2-P2O5-B2O3 glass-ceramics with high osteoconductivity that had enhanced resorption by having an increased B2O3 content. The three types of CaO-SiO2-P2O5-B2O3 glass-ceramics with B2O3 contents of 8.0, 9.0, and 9.5 weight % were designated and grouped as P20B80, P10B90, and P5B95, respectively. Glass-ceramic types were tested for fusion rates and bone formation by employing the lumbar 5-6 intertransverse process fusion model in 51 New Zealand male rabbits. Bioactivity was assessed by soaking in simulated body fluid (SBF).ResultsIn vitro study results showed sufficient hydroxycarbonate apatite layer formation occurred for P20B80 in1 day, for P10B90 in 3 days, and for P5B95 in 5 days after soaking in SBF. For the rabbit lumbar spine posterolateral fusion model, the autograft group recorded a 100% fusion rate with levels significantly higher than those of P20B80 (29.4%), P10B90 (0%), and P5B95 (14.3%), with high resorbing properties. Resorbing property differences among the three glass-ceramic groups were not significant. Histological results showed new bone formation confirming osteoconductivity in all three types of glass-ceramics. Radiomorphometric results also confirmed the resorbing properties of the three glass-ceramic types.ConclusionsThe high resorbing properties and osteoconductivity of porous glass-ceramics can be advantageous as no glass-ceramics remain in the body. However, their relatively fast rate of resorption in the body negatively affects their role as an osteoconductive scaffold as glass-ceramics are resorbed before bony fusion.

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.

Repeat Use of Human Recombinant Bone Morphogenetic Protein-2 for Second Level Lumbar Arthrodesis

Prospective randomized controlled animal model. The purpose of this study is to determine whether the readministration of human recombinant bone morphogenetic protein-2 (rhBMP-2) induces an immune response and inhibits successful fusion in repeat posterolateral spinal surgery. Little research has been performed on the effectiveness or immunoreactivity of rhBMP-2 (Infuse, Medtronic, Memphis, TN) in the context of its reuse in posterolateral fusion spinal surgery at adjacent levels. A total of 34 New Zealand White rabbits underwent posterior intertransverse process fusion with the use of rhBMP-2 delivered on an absorbable collagen sponge (rhBMP-2/ACS). Two rabbits were killed early leaving 32 total rabbits. Serologic studies (Type I bovine collagen and rhBMP-2 antibodies) were obtained at 2-week intervals throughout the experiment. At 10 weeks, posteroanterior radiographs confirmed solid fusion masses in all rabbits. The 32 rabbits were randomly separated into 2 groups of 16, and each group underwent an adjacent level, bilateral intertransverse process fusion with either rhBMP-2/ACS or iliac crest. There was no statistical difference in fusion rates with repeat use of rhBMP-2 (n = 15/16, 94%) or iliac crest (n = 11/16, 69%) (P = 0.17) at the adjacent level. Four rabbits (n = 4/32, 13%) developed rhBMP-2 antibodies. Of these 4 rabbits, 1 developed anti-rhBMP antibodies after the first exposure and 3 developed antibodies after the second surgery. Eight rabbits (n = 8/32, 25%) developed collagen antibodies with 7 rabbits developing antibodies after the first exposure and 1 rabbit developing antibodies after the second exposure. The development of antibodies did not effect fusion rates. No rabbit demonstrated evidence of a systemic or anaphylactic reaction to repeat exposure to rhBMP-2. rhBMP-2 appears to be successful in promoting intertransverse fusions when used in both primary and repeat fusion environments. The infrequent development of antibodies to rhBMP-2 after re-exposure occurs without a predictable time course suggesting that host immunologic variation may play a role. This animal study would tend to support early clinical data emerging on the reuse of BMP-2 for lumbar fusion, suggesting an acceptable fusion rate without a high incidence of complications.

Biological evaluation of recombinant human bone morphogenetic protein-2 artificial bone in lumbar intertransverse process fusion

Objective To evaluate rhBMP-2/20%β-TCP/PDLLA complex biological properties.Methods Preparation of casting salting was carried out using three-dimensional porous 20% rhBMP-2/β-TCP/PDLLA. 24 4-month-old healthy male New Zealand rabbits were randomly divided into A,B groups of 12,according to the method described in Boden production L4,5 intertransverse process fusion model. A group was embedding rhBMP-2/20%β-TCP/PDLLA complex,and B group was implanted autogenous iliac bone block. Animals were killed according to postoperative time(3 weeks,6 weeks, 12 weeks) ,for gross observation, X-ray observation of the formation of new bone, artificial bone degradation, and early histological manifestations. Results Composite artificial bone and autologous iliac bone graft were able to achieve good spinal fusion,artificial bone graft where there is no inflammatory response,but also in new bone formation,while reshaping the gradual degradation of carrier material. Conclusions 20%β-TCP/PDLLA showed good biocompatibility. Its structure and micro-environment created by its degradation were conducive to play a role for the rhBMP-2. β-TCP/PDLLA had strong activity in inducing bone and good bone fusion ability,and it was a promising bone graft substitutes, which fusion power was stronger than that in autologous iliac bone graft. Key words: Bone morphogenetic porteins; Bone substitutes; Spinal fractures; Biomechanics

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.

Involvement of Calcitonin Gene-Related Peptide Innervation in the Promoting Effect of Low-Intensity Pulsed Ultrasound on Spinal Fusion Without Decortication

A prospective left-right comparison designed experiment using a rabbit posterolateral intertransverse process fusion model. To investigate the involvement of calcitonin gene-related peptide (CGRP) innervation in the promoting effect of low intensity pulsed ultrasound stimulation (LIPUS) on spinal fusion without decortication. Sensory neuropeptide CGRP is involved in bone repair and ectopic ossification. Comparison of CGRP innervations in ectopic bone between sham LIPUS and LIPUS sides can help us to understand the relationship between sensory nerve innervation and LIPUS. A total of 27 New Zealand white rabbits underwent bilateral posterolateral intertransverse process fusion with implantation of porous poly-D,L-lactic acid blocks loaded with 1.25 μg recombinant human bone morphogenetic protein-4 solution. One side was provided LIPUS daily whereas the other side served as control. Animals were killed and the operated lumbar vertebrae were harvested for histomorphologic evaluation at 3 days (n = 3), 1 week (n = 6), 3 weeks (n = 6), 7 weeks (n = 6), and 12 weeks (n = 6) following surgery, respectively. LIPUS accelerated the invasion of CGRP-positive nerve fibers during ectopic ossification spatially and temporally. Spatially, CGRP-positive nerve fibers were also observed in the new formed cartilage and bone tissues on LIPUS side, whereas they were only detected in the fibrous tissue and bone marrow on sham LIPUS side. Temporally, the density of CGRP-positive nerve fibers was significantly higher on the LIPUS side when compared with the sham LIPUS side. LIPUS promoted the invasion of CGRP sensory nerve in ectopic bone, which may in turn contribute to the promoting effect of LIPUS on ectopic ossification.

Positron emission tomography-CT study on posterolateral lumbar spinal fusion with recombinant human bone morphogenetic protein-2/allograft composite in a rabbit model

Objective To investigate revascularization and osteogenesis of the fusion masses in rabbits undergoing posterolateral lumbar spinal fusion with recombinant human bone morphogenetic protein-2 (rhBMP-2)/allograft composite. Methods Forty-five adult male New Zealand white rabbits were randomly divided into 3 even groups. The rabbits in each group undergoing intertransverse process fusion at L4 and L5 respectively with rhBMP-2/allograft composite (group A), autograft (group B) and allograft (group C). 18F-NaF was injected into every rat at 2nd, 4th and 6th weeks after surgery. Whole-body radionuclide scan by PET-CT was done for all rats to evaluate the uptake values (SU V) in fusion areas. Results The SUVs for autograft and composite groups were superior to that for allograft group at 2nd, 4th and 6th weeks ( P ＜0. 05). The SUV for composite group was significantly higher than that for autograft group at the 4th and 6th weeks ( P ＜0. 05), but similar to that for autograft group at the 2nd week ( P ＞ 0. 05) . Conclusion Since the rhBMP-2/allograft composite can not only promote the bone formation but also improve the blood supply in the posterolateral lumbar spinal fusion, it may serve as a substitute material for autografts. Key words: Bone morphogenetic proteirs; Spinal fusion; Lumbar vertebrae; Tomography,spiral computed