A comparison of the inflammatory host response to particulate debris adjacent to unlocked and locked screws of a growth guidance system for early onset scoliosis.

PURPOSE: The SHILLA™ Growth Guidance system is a stainless-steel rod and screw system used for Early Onset Scoliosis which incorporates a unique flanged set screw designed to capture the rod, while allowing it to slide as the patient grows. Concomitant with this design is the potential for generation of wear debris and for an inflammatory host response. We hypothesized that the magnitude of the host response adjacent to the unlocked screws and rods would be greater than the host response to the locked rod/screws. METHODS: Seven tissue samples adjacent to locked (3) and unlocked screws (4) from three SHILLA patients (mean implantation time of 19 post-operative months) with infantile idiopathic scoliosis were obtained as part of an explant analysis protocol during a PMDA-approved clinical trial in Japan. Gross appearance, high-resolution radiographs, and histology were assessed. ISO Standard 10993 Part 6 was used to assess the host response. RESULTS: All three locked screw had no metallosis. In contrast, metallosis for unlocked screw tissue samples were rated as "ubiquitous" (2/4), "focal" (1/4), or "absent" (1/4). Microscopic metallic debris was found intracellularly and within interstices of fibrous connective tissues more frequently adjacent to unlocked screws compared to locked screws. Cell type and population scoring consistently showed a modestly larger inflammatory response (macrophages) in the unlocked tissue samples. CONCLUSIONS: The peri-prosthetic tissue response to the unlocked rods/screws had a higher reactivity grade (slight reaction, Δ = 4.0) per ISO 10993 Part 6 compared to the locked screws in three patients with the SHILLA™ Growth Guidance scoliosis system.

Alagebrium inhibits neointimal hyperplasia and restores distributions of wall shear stress by reducing downstream vascular resistance in obese and diabetic rats.

Mechanisms of restenosis in type 2 diabetes mellitus (T2DM) are incompletely elucidated, but advanced glycation end-product (AGE)-induced vascular remodeling likely contributes. We tested the hypothesis that AGE-related collagen cross-linking (ARCC) leads to increased downstream vascular resistance and altered in-stent hemodynamics, thereby promoting neointimal hyperplasia (NH) in T2DM. We proposed that decreasing ARCC with ALT-711 (Alagebrium) would mitigate this response. Abdominal aortic stents were implanted in Zucker lean (ZL), obese (ZO), and diabetic (ZD) rats. Blood flow, vessel diameter, and wall shear stress (WSS) were calculated after 21 days, and NH was quantified. Arterial segments (aorta, carotid, iliac, femoral, and arterioles) were harvested to detect ARCC and protein expression, including transforming growth factor-ß (TGF-ß) and receptor for AGEs (RAGE). Downstream resistance was elevated (60%), whereas flow and WSS were significantly decreased (44% and 56%) in ZD vs. ZL rats. NH was increased in ZO but not ZD rats. ALT-711 reduced ARCC and resistance (46%) in ZD rats while decreasing NH and producing similar in-stent WSS across groups. No consistent differences in RAGE or TGF-ß expression were observed in arterial segments. ALT-711 modified lectin-type oxidized LDL receptor 1 but not RAGE expression by cells on decellularized matrices. In conclusion, ALT-711 decreased ARCC, increased in-stent flow rate, and reduced NH in ZO and ZD rats through RAGE-independent pathways. The study supports an important role for AGE-induced remodeling within and downstream of stent implantation to promote enhanced NH in T2DM.

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: implications for bone graft preparation.

STUDY DESIGN: 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. OBJECTIVE: 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. SUMMARY OF BACKGROUND DATA: 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. METHODS: Twelve skeletally mature, rhesus macaque monkeys underwent single-level posterolateral arthrodesis at L4-L5. A hydroxyapatite/ß-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. RESULTS: 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. CONCLUSIONS: 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.

Development of fibroblast/endothelial cell-seeded collagen scaffolds for in vitro prevascularization.

The development of vascularized scaffolds remains one of the major challenges in tissue engineering, and co-culturing with endothelial cells is known as one of the possible approaches for this purpose. In this approach, optimization of cell culture conditions, scaffolds, and fabrication techniques is needed to develop tissue equivalents that will enable in vitro formation of a capillary network. Prevascularized equivalents will be more physiologically comparable to the native tissues and potentially prevent insufficient vascularization after implantation. This study aimed to culture human umbilical vein endothelial cells (HUVECs), alone or in co-culture with fibroblasts, on collagen scaffolds prepared by simple fabrication approaches for in vitro prevascularization. Different concentrations and ratios of HUVECs and fibroblasts seeded on collagen gel and sponge scaffolds under several culture conditions were examined. Cell viability, scaffolds morphology, and structure were analyzed. Collagen gel scaffolds showed good cell proliferation and viability, with higher proliferation rates for cells cultured in a 2:1 (fibroblasts: HUVECs) ratio and kept in endothelial cell growth medium. However, these matrices were unable to support endothelial cell sprouting. Collagen sponges were highly porous and showed good cell viability. However, they became fragile over time in culture, and they still lack signs of vascularization. Collagen scaffolds were a good platform for cell growth and viability. However, under the experimental conditions of this study, the HUVEC/fibroblast-seeded scaffolds were not suitable platforms to generate in vitro prevascularized equivalents. Our findings will be a valuable starting point to optimize culture microenvironments and scaffolds during fabrication of prevascularized scaffolds.

Altered hemodynamics, endothelial function, and protein expression occur with aortic coarctation and persist after repair.

Coarctation of the aorta (CoA) is associated with substantial morbidity despite treatment. Mechanically induced structural and functional vascular changes are implicated; however, their relationship with smooth muscle (SM) phenotypic expression is not fully understood. Using a clinically representative rabbit model of CoA and correction, we quantified mechanical alterations from a 20-mmHg blood pressure (BP) gradient in the thoracic aorta and related the expression of key SM contractile and focal adhesion proteins with remodeling, relaxation, and stiffness. Systolic and mean BP were elevated for CoA rabbits compared with controls leading to remodeling, stiffening, an altered force response, and endothelial dysfunction both proximally and distally. The proximal changes persisted for corrected rabbits despite >12 wk of normal BP (~4 human years). Computational fluid dynamic simulations revealed reduced wall shear stress (WSS) proximally in CoA compared with control and corrected rabbits. Distally, WSS was markedly increased in CoA rabbits due to a stenotic velocity jet, which has persistent effects as WSS was significantly reduced in corrected rabbits. Immunohistochemistry revealed significantly increased nonmuscle myosin and reduced SM myosin heavy chain expression in the proximal arteries of CoA and corrected rabbits but no differences in SM α-actin, talin, or fibronectin. These findings indicate that CoA can cause alterations in the SM phenotype contributing to structural and functional changes in the proximal arteries that accompany the mechanical stimuli of elevated BP and altered WSS. Importantly, these changes are not reversed upon BP correction and may serve as markers of disease severity, which explains the persistent morbidity observed in CoA patients.

The Latest Lessons Learned from Retrieval Analyses of Ultra-High Molecular Weight Polyethylene, Metal-on-Metal, and Alternative Bearing Total Disc Replacements.

Knowledge regarding the in vivo performance and periposthetic tissue response of cervical and lumbar total disc replacements (TDRs) continues to expand. This review addresses the following four main questions: 1) What are the latest lessons learned from polyethylene in large joints and how are they relevant to current TDRs? 2) What are the latest lessons learned regarding adverse local tissue reactions from metal-on-metal, CoCr bearings in large joints and how are they relevant to current TDRs? 3) What advancements have been made in understanding the in vivo performance of alternative biomaterials, such as stainless steel and polycarbonate urethane, for TDRs in the past five years? 4) How has retrieval analysis of all these various artificial disc bearing technologies advanced the state of the art in preclinical testing of TDRs? The study of explanted artificial discs and their associated tissues can help inform bearing selection as well as the design of future generations of disc arthroplasty. Analyzing retrieved artificial discs is also essential for validating preclinical test methods.

Biological resurfacing in a canine model of hip osteoarthritis.

Articular cartilage has unique load-bearing properties but has minimal capacity for intrinsic repair. Here, we used three-dimensional weaving, additive manufacturing, and autologous mesenchymal stem cells to create a tissue-engineered, bicomponent implant to restore hip function in a canine hip osteoarthritis model. This resorbable implant was specifically designed to function mechanically from the time of repair and to biologically integrate with native tissues for long-term restoration. A massive osteochondral lesion was created in the hip of skeletally mature hounds and repaired with the implant or left empty (control). Longitudinal outcome measures over 6 months demonstrated that the implant dogs returned to normal preoperative values of pain and function. Anatomical structure and functional biomechanical properties were also restored in the implanted dogs. Control animals never returned to normal and exhibited structurally deficient repair. This study provides clinically relevant evidence that the bicomponent implant may be a potential therapy for moderate hip osteoarthritis.

An evaluation of the host response to an interspinous process device based on a series of spine explants: Device for Intervertebral Assisted Motion (DIAM®).

BACKGROUND: The objective of this study was to evaluate the host response to an interspinous process device [Device for Intervertebral Assisted Motion (DIAM®)] based on a series of nine spine explants with a mean post-operative explant time of 35 months. METHODS: Explanted periprosthetic tissues were processed for histology and stained with H&E, Wright-Giemsa stain, and Oil Red O. Brightfield and polarized light microscopy were used to evaluate the host response to the device and the resultant particulate debris. The host response was graded per ASTM F981-04. Quantitative histomorphometry was used to characterize particle size, shape, and area per ASTM F1877-05. The presence or absence of bone resorption was also evaluated when bony tissue samples were provided. RESULTS: Periprosthetic tissues demonstrated a non-specific foreign body response composed of macrophages and foreign body giant cells to the DIAM® device in most of the accessions. The foreign body reaction was not the stated reason for explantation in any of the accessions. Per ASTM F981-04, a "very slight" to "mild" to "moderate" chronic inflammatory response was observed to the biomaterials and particulate, and this varied by tissue sample and accession. Particle sizes were consistent amongst the explant patients with mean particle size on the order of several microns. Osteolysis, signs of toxicity, necrosis, an immune response, and/or device related infection were not observed. CONCLUSIONS: Cyclic loading of the spine can cause wear in dynamic stabilization systems such as DIAM®. The fabric nature of the DIAM® device's polyethylene terephthalate jacket coupled with the generation of polymeric particulate debris predisposes the device to a foreign body reaction consisting of macrophages and foreign body giant cells. Although not all patients are aware of symptoms associated with a foreign body reaction to a deeply implanted device, surgeons should be aware of the host response to this device.

Mineral trioxide aggregate material use in endodontic treatment: a review of the literature.

OBJECTIVE: The purpose of this paper was to review the composition, properties, biocompatibility, and the clinical results involving the use of mineral trioxide aggregate (MTA) materials in endodontic treatment. METHODS: Electronic search of scientific papers from January 1990 to August 2006 was accomplished using PubMed and Scopus search engines (search terms: MTA, GMTA, WMTA, mineral AND trioxide AND aggregate). RESULTS: Selected exclusion criteria resulted in 156 citations from the scientific, peer-reviewed dental literature. MTA materials are derived from a Portland cement parent compound and have been demonstrated to be biocompatible endodontic repair materials, with its biocompatible nature strongly suggested by its ability to form hydroxyappatite when exposed to physiologic solutions. With some exceptions, MTA materials provide better microleakage protection than traditional endodontic repair materials using dye, fluid filtration, and bacterial penetration leakage models. In both animal and human studies, MTA materials have been shown to have excellent potential as pulp-capping and pulpotomy medicaments but studies with long-term follow-up are limited. Preliminary studies suggested a favorable MTA material use as apical and furcation restorative materials as well as medicaments for apexogenesis and apexification treatments; however, long-term clinical studies are needed in these areas. CONCLUSION: MTA materials have been shown to have a biocompatible nature and have excellent potential in endodontic use. MTA materials are a refined Portland cement material and the substitution of Portland cement for MTA products is presently discouraged. Existing human studies involving MTA materials are very promising, however, insufficient randomized, double-blind clinical studies of sufficient duration exist involving MTA for all of its clinical indications. Further clinical studies are needed in these areas.

Early term effects of rhBMP-2 on pedicle screw fixation in a sheep model: histomorphometric and biomechanical analyses.

BACKGROUND: The effects of recombinant human bone morphogenetic protein-2 (rhBMP-2) on pedicle screw pullout force and its potential to improve spinal fixation have not previously been investigated. rhBMP-2 on an absorbable collagen sponge (ACS) carrier was delivered in and around cannulated and fenestrated pedicle screws in a sheep lumbar spine instability model. Two control groups (empty screw and ACS with buffer) were also evaluated. We hypothesized that rhBMP-2 could stimulate bone growth in and around the cannulated and fenestrated pedicle screws to improve early bone purchase. METHODS: Eight skeletally mature sheep underwent destabilizing laminectomies at L2-L3 and L4-L5 followed by stabilization with pedicle screw and rod constructs. An ACS carrier was used to deliver 0.15 mg of rhBMP-2 within and around the cannulated and fenestrated titanium pedicle screws. Biomechanics and histomorphometry were used to evaluate the early term results at 6 and 12 postoperative weeks. RESULTS: rhBMP-2 was unable to improve bony purchase of the cannulated and fenestrated pedicle screws compared to both control groups. Although rhBMP-2 groups had pullout forces that were less than both control groups, both rhBMP-2 groups had pullout force values exceeding 2,000 N, which was comparable to previously published results for unmodified pedicle screws. Significant differences in the percentages of bone in peri-screw tissues was not observed amongst the four treatment groups. Microradiography and quantitative histomorphometry showed that at 6 weeks, rhBMP-2 induced peri-screw remodeling regions containing peri-implant bone which was hypodense with respect to surrounding native trabeculae. A moderate correlation between biomechanical pullout variables and histomorphometry data was observed. CONCLUSIONS: The design of the cannulated and fenestrated pedicle screw was able to facilitate new bone formation to achieve high pullout forces. However, delivery of rhBMP-2 should be carefully controlled to prevent excessive bone remodeling which could cause early screw loosening.

Opioids delay healing of spinal fusion: a rabbit posterolateral lumbar fusion model.

BACKGROUND CONTEXT: Opioid use is prevalent in the management of pre- and postoperative pain in patients undergoing spinal fusion. There is evidence that opioids downregulate osteoblasts in vitro, and a previous study found that morphine delays the maturation and remodeling of callus in a rat femur fracture model. However, the effect of opioids on healing of spinal fusion has not been investigated before. Isolating the effect of opioid exposure in humans would be limited by the numerous confounding factors that affect fusion healing. Therefore, we have used a well-established rabbit model to study the process of spinal fusion healing that closely mimics humans. PURPOSE: The objective of this work was to study the effect of systemic opioids on the process of healing of spinal fusion in a rabbit posterolateral spinal fusion model. STUDY DESIGN/SETTING: This is a preclinical animal study. MATERIALS AND METHODS: Twenty-four adult New Zealand white rabbits were studied in two groups after approval from the Institutional Animal Care and Use Committee (IACUC). The opioid group (n=12) received 4 weeks' preoperative and 6 weeks' postoperative transdermal fentanyl. Serum fentanyl levels were measured just before surgery and 4 weeks postoperatively to ensure adequate levels. The control group (n=12) received only perioperative pain control as necessary. All animals underwent a bilateral L5-L6 posterolateral spinal fusion using iliac crest autograft. Animals were euthanized at the 6-week postoperative time point, and assessment of fusion was done by manual palpation, plain radiographs, microcomputed tomography (microCT), and histology. RESULTS: Twelve animals in the control group and 11 animals in the opioid group were available for analysis at the end of 6 weeks. The fusion scores on manual palpation, radiographs, and microCT were not statistically different. Three-dimensional microCT morphometry found that the fusion mass in the opioid group had a lower bone volume (p=.09), a lower trabecular number (p=.02), and a higher trabecular separation (p=.02) compared with the control group. Histologic analysis found areas of incorporation of autograft and unincorporated graft fragments in both groups. In the control group, there was remodeling of de novo woven bone to lamellar organization with incorporation of osteocytes, formation of mature marrow, and relative paucity of hypertrophied osteoblasts lining new bone. Sections from the opioid group showed formation of de novo woven bone, and hypertrophied osteoblasts were seen lining the new bone. There were no sections showing lamellar organization and development of mature marrow elements in the opioid group. Less dense trabeculae on microCT correlated with histologic findings of relatively immature fusion mass in the opioid group. CONCLUSIONS: Systemic opioids led to an inferior quality fusion mass with delay in maturation and remodeling at 6 weeks in this rabbit spinal fusion model. These preliminary results lay the foundation for further research to investigate underlying cellular mechanisms, the temporal fusion process, and the dose-duration relationship of opioids responsible for our findings.

The temporal expression of adipokines during spinal fusion.

BACKGROUND CONTEXT: Adipokines are secreted by white adipose tissue and have been associated with fracture healing. Our goal was to report the temporal expression of adipokines during spinal fusion in an established rabbit model. PURPOSE: Our goal was to report the temporal expression of adipokines during spinal fusion in an established rabbit model. STUDY DESIGN: The study design included a laboratory animal model. METHODS: New Zealand white rabbits were assigned to either sham surgery (n=2), unilateral posterior spinal fusion (n=14), or bilateral posterior spinal fusion (n=14). Rabbits were euthanized 1-6 and 10 weeks out from surgery. Fusion was evaluated by radiographs, manual palpation, and histology. Reverse transcription-polymerase chain reaction on the bone fusion mass catalogued the gene expression of leptin, adiponectin, resistin, and vascular endothelial growth factor (VEGF) at each time point. Results were normalized to the internal control gene, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (2^ΔCt), and control bone sites (2^ΔΔCt). Quantitative data were analyzed by two-factor analysis of variance (p<.05). RESULTS: Manual palpation scores, radiograph scores, and histologic findings showed progression of boney fusion over time (p<.0003). The frequency of fusion by palpation after 4 weeks was 68.75%. Leptin expression in decortication and bone graft sites peaked at 5 weeks after the fusion procedure (p=.0143), adiponectin expression was greatest 1 week after surgery (p<.001), VEGF expression peaked at 4 weeks just after initial increases in leptin expression (p<.001), and resistin decreased precipitously 1 week after the fusion procedure (p<.001). CONCLUSIONS: Leptin expression is likely associated with the maturation phase of bone fusion. Adiponectin and resistin may play a role early on during the fusion process. Our results suggest that leptin expression may be upstream of VEGF expression during spinal fusion, and both appear to play an important role in bone spinal fusion.

Is lumbar facet fusion biomechanically equivalent to lumbar posterolateral onlay fusion?

OBJECTIVE This study was designed with the following research objectives: 1) to determine the efficacy of facet fusion with recombinant human bone morphogenetic protein-2 (rhBMP-2) on an absorbable collagen sponge (ACS) in an ovine lumbar facet fusion model; 2) to radiographically and histologically compare the efficacy of lumbar facet fusion with rhBMP-2/ACS to facet fusion with an iliac crest bone graft (ICBG); and 3) to biomechanically compare lumbar facet fusion with rhBMP-2/ACS to lumbar posterolateral fusion (PLF) with ICBG. METHODS The efficacies of the 3 treatments to induce fusion were evaluated in an instrumented ovine lumbar fusion model. Eight sheep had 10 cm3/side ICBG placed as an onlay graft for PLF at L2-3. At the adjacent L3-4 level, 0.5 cm3/side ICBG was placed for facet fusion. Finally, 0.5 cm3/side rhBMP-2/ACS (0.43 mg/ml) was placed for facet fusion at L4-5. CT scans were obtained at 2, 4, and 6 months postoperatively with 2 reviewers conducting an evaluation of the 6-month results for all treated spinal levels. All 8 sheep were killed at 6 months, and all posterolateral instrumentation was removed at this time. The spines were then sectioned through L3-4 to allow for nondestructive unconstrained biomechanical testing of the L2-3 and L4-5 segments. All treated spinal levels were analyzed using undecalcified histology with corresponding microradiography. Statistical comparisons were made between the treatment groups. RESULTS The PLF with ICBG (ICBG PLF group) and the rhBMP-2 facet fusion (rhBMP-2 Facet group) treatment groups demonstrated similar levels of stiffness, with the rhBMP-2 Facet group having on average slightly higher stiffness in all 6 loading directions. All 8 levels in the autograft facet fusion treatment group demonstrated CT radiographic and histological fusion. All 8 levels in the rhBMP-2 Facet group showed bilateral CT radiographic and histological fusion. Six of 16 rhBMP-2/ACS-treated facet defects demonstrated small intraosseous hematomas or seromas. Four of the 8 levels (50%) in the ICBG PLF treatment group demonstrated bilateral histological fusion. Three of 8 levels in the ICBG PLF treatment group showed unilateral fusion. One of the 8 levels in the ICBG PLF treatment group demonstrated bilateral histological nonfusion. CONCLUSIONS Both rhBMP-2/ACS and autograft demonstrated 100% efficacy when used for facet fusion in the instrumented ovine model. However, the ICBG PLF treatment group only demonstrated a 50% bilateral fusion rate. Biomechanically, the ICBG PLF and rhBMP-2 Facet groups demonstrated similar stiffness in all 6 loading directions, with the rhBMP-2 Facet group having on average slightly higher stiffness in all directions.

Polyetheretherketone as a biomaterial for spinal applications.

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

Effect on bone induction of using contrast media to reconstitute recombinant human bone morphogenetic protein-2 in an ectopic model in rats.

OBJECT: In this study the authors tested the osteoinductive potential of recombinant human bone morphogenetic protein-2 (rhBMP-2) when combined with each of three commercially available contrast media (Conray, Omniscan, and Optiray). METHODS: Initial in vitro and cadaver tests verified the feasibility of using contrast media to visualize absorbable collagen sponge implants containing rhBMP-2 on fluoroscopic radiographic images. For the feasibility studies, lyophilized rhBMP-2 was prepared for injection by reconstitution with contrast media instead of sterile water. For the in vivo study, samples of an rhBMP-2 stock solution were diluted to 0.1 mg/ml by using three contrast media. In each sample, the final solution consisted of 97% contrast medium by volume. Recombinant human bone morphogenetic protein-2 diluted with sterile water for injection was used as a positive control. The rhBMP-2 solutions were applied to 0.5-cm3 collagen sponges and implanted subcutaneously on the thoracic cavity of athymic rats. At 4 weeks, the rats were killed, and the implants were removed. The explants were graded for degree of bone formation by using manual palpation and radiographic and histological assessments. CONCLUSIONS: By all methods of evaluation used, rhBMP-2 diluted with Omniscan was equivalent to rhBMP-2 diluted with sterile water in inducing bone formation. Both Conray and Optiray were shown to inhibit the osteoinductive potential of rhBMP-2.

Instrumented Lumbar Corpectomy and Spinal Reconstruction Comparing rhBMP-2/Compression-Resistant Matrix, rhBMP-2/Absorbable Collagen Sponge/Ceramic Granules Mixture, and Autograft in Two Different Devices: A Study in Sheep.

STUDY DESIGN: Fusion success with rhBMP-2 and autograft in titanium or PEEK corpectomy devices was evaluated in a sheep lumbar corpectomy model. The 6 treatment groups included titanium mesh or PEEK corpectomy devices filled with rhBMP-2 on a compression-resistant matrix (CRM) carrier; rhBMP-2 in a morselized absorbable collagen sponge (ACS) carrier combined with resorbable ceramic granules; and autograft. OBJECTIVE: The aim of this study was to determine fusion rates associated with 2 different preparations of rhBMP-2 as well as autograft in an instrumented ovine lumbar corpectomy model 6 months postoperatively. SUMMARY OF BACKGROUND DATA: Vertebral reconstruction with corpectomy devices requires bone graft. Bone graft substitutes have the potential to avoid a second operation, donor site pain, and attendant morbidity associated with autograft. METHODS: Twenty-four sheep in 6 treatment groups underwent lumbar corpectomy via a retroperitoneal trans-psoas approach. Spines were reconstructed with autograft, rhBMP-2 on a CRM, or rhBMP-2 on an ACS mixed with ceramic granules. Grafting materials were placed in either a titanium mesh or PEEK conduit in spines with internal fixation. Computed tomographic (CT) scans were evaluated for fusion. Undecalcified histology was used to evaluate for fusion as well as the amount and extent of graft incorporation and graft resorption. RESULTS: Regardless of corpectomy device used, rhBMP-2/CRM or rhBMP-2/ACS with MASTERGRAFT resulted in a 100% fusion rate. The autograft group had a lower (75%) radiographic fusion rate. Using either preparation of rhBMP-2 resulted in the length of the defect filling with solid bone. Autograft fragments and ceramic granules were incorporated into the fusion masses with much of the ceramic granules being resorbed by 6 months. CONCLUSION: Both of the rhBMP-2 formulations have the potential to effect bony fusion and vertebral reconstruction within the corpectomy devices.

Radiographic, biomechanical, and histological evaluation of rhBMP-2 in a 3-level intertransverse process spine fusion: an ovine study.

OBJECTIVE The objective of this study was to evaluate bone grafts consisting of rhBMP-2 on an absorbable collagen sponge with a ceramic composite bulking agent, rhBMP-2, directly on a ceramic-collagen sponge carrier or iliac crest bone graft (ICBG) in combination with local bone graft to effect fusion in a multisegmental instrumented ovine lumbar intertransverse process fusion model. METHODS Thirty-six sheep had a single treatment at 3 spinal levels in both the right and left intertransverse process spaces. Group 1 sheep were treated with 7.5 cm3 of autograft consisting of ICBG plus local bone for each intertransverse process space. For Groups 2-4, 4 cm3 of local bone was placed within the intertransverse process space followed by 4.5-5 cm3 of the rhBMP-2 graft material. Group 2 animals received 1.5 mg/cm3 rhBMP-2 on an absorbable collagen sponge with a commercial bone void filler consisting of Type I lyophilized collagen with a biphasic hydroxyapatite/ß-tricalcium phosphate ceramic with local bone. Group 3 animals received 0.75 mg/m cm3 of rhBMP-2 on a collagen ceramic sponge carrier with local bone. Group 4 animals received 1.35 mg/cm3 of rhBMP-2 on the same collagen ceramic sponge carrier with local bone. Sheep were euthanized 6 months postoperatively. Manual palpation, biomechanical testing, CT, radiography, and undecalcified histology were performed to assess the presence of fusion associated with the treatments. RESULTS All animals in Groups 2-4 that received grafts containing rhBMP-2 achieved radiographic and CT fusion at all 3 levels. In Group 1 (bone autograft alone), only 19% of the levels demonstrated radiographic fusion, 14% resulted in possible radiographic fusion, and 67% of the levels demonstrated radiographic nonfusion. Biomechanical testing showed that Groups 2-4 demonstrated similar stiffness of the L2-5 segment in all 6 loading directions, with each of the 3 groups having significantly greater stiffness than the autograft-only group. In Group 1, only 2 of 18 levels were rated as achieving bilateral histological fusion, with an additional 3 levels showing a unilateral fusion. The majority of the treated levels (13/18) in Group 1 were scored as histological nonfusions. There were no histological nonfusions in Groups 2 through 4. All 18 levels in Group 2 were rated as bilateral histological fusions. A majority (34/36) of the levels in Group 3 were rated as bilateral histological fusions, with 2 levels showing a unilateral fusion. A majority (35/36) of the levels in Group 4 were rated as bilateral histological fusions, with 1 level showing a unilateral fusion. CONCLUSIONS In the ovine multilevel instrumented intertransverse process fusion model, rhBMP-2 was able to consistently achieve CT, radiographic, biomechanical, and histological fusion. Compared with ICBG, the gold standard for bone grafting, rhBMP-2 was statistically superior at achieving radiographic and histological fusion.

Transient Local Bone Remodeling Effects of rhBMP-2 in an Ovine Interbody Spine Fusion Model.

BACKGROUND: Recombinant human bone morphogenetic protein-2 (rhBMP-2) is a powerful osteoinductive morphogen capable of stimulating the migration of mesenchymal stem cells (MSCs) to the site of implantation and inducing the proliferation and differentiation of these MSCs into osteoblasts. Vertebral end-plate and vertebral body resorption has been reported after interbody fusion with high doses of rhBMP-2. In this study, we investigated the effects of 2 rhBMP-2 doses on peri-implant bone resorption and bone remodeling at 7 time points in an end-plate-sparing ovine interbody fusion model. METHODS: Twenty-one female sheep underwent an end-plate-sparing discectomy followed by interbody fusion at L2-L3 and L4-L5 using a custom polyetheretherketone (PEEK) interbody fusion device. The PEEK interbody device was filled with 1 of 2 different doses of rhBMP-2 on an absorbable collagen sponge (ACS): 0.13 mg (1×) or 0.90 mg (7×). Bone remodeling and interbody fusion were assessed via high-resolution radiography and histological analyses at 1, 2, 3, 4, 8, 12, and 20 weeks postoperatively. RESULTS: Peri-implant bone resorption peaked between 3 and 8 weeks in both the 1× and the 7× rhBMP-2/ACS-dose group. Osteoclastic activity and corresponding peri-implant bone resorption was dose-dependent, with moderate-to-marked resorption at the 7×-dose level and less resorption at the 1×-dose level. Both dose (p < 0.0007) and time (p < 0.0025) affected bone resorption significantly. Transient bone-resorption areas were fully healed by 12 weeks. Osseous bridging was seen at all but 1 spinal level at 12 and at 20 weeks. CONCLUSIONS: In the ovine end-plate-sparing interbody fusion model, rhBMP-2 dose-dependent osteoclastic resorption is a transient phenomenon that peaks at 4 weeks postoperatively. CLINICAL RELEVANCE: Using the U.S. Food and Drug Administration (FDA)-approved rhBMP-2 concentration and matching the volume of rhBMP-2/ACS with the volume of desired bone formation within the interbody construct may limit the occurrence of transient bone resorption.

An in vitro comparison of the rake angles between K3 and ProFile endodontic file systems.

The purpose of this study was to compare rake angles of the ProFile and K3 file systems. Twenty-five 40/0.06 taper files were obtained for each system. Five files from the same manufacturer were placed perpendicularly into a vial of Epoxicure Resin and left to set for 24 h. The set-ups were removed from the vials and each were sectioned 5 mm from the tip of the files and polished. A photomicrograph was taken of each file with 100x magnification. Five sets of ProFile and five sets of K3 files were processed in this manner. Images were captured digitally, and rake angles of each file were measured. Multivariate ANOVA found a significant difference (p < 0.001) among the three negative rake angles of the ProFile system compared with the K3 system.

Use of polylactide resorbable film as an adhesion barrier.

OBJECT: The present investigation evaluates two thicknesses of a resorbable polylactic acid (PLA) barrier film as an adhesion barrier to posterior spinal scar formation. METHODS: A readily contourable, thin film was placed directly over the dura. The thick film was placed above the lamina defect to act as a physical barrier inhibiting the prolapse of soft tissue into the epidural space. Through a combination of gross dissection with and without scar scores, quantitative analysis of collagen adjacent to the scar site, and histological evaluation, the resorbable adhesion barrier membranes were found to be effective treatment for reduction of posterior adhesions. CONCLUSIONS: The gross dissection demonstrated that both thicknesses of resorbable PLA barrier films created a controlled dissection plane, facilitated access to the epidural space, and provided a reduction in the tissue adherent to the dura.