RESUMO
STUDY DESIGN: Assessment of bone formation in an ovine interbody fusion study. OBJECTIVE: To compare OsteoAdapt SP, which consists of AMP-2, a modified variant of recombinant human bone morphogenetic protein (rhBMP-2) bound to a tricalcium phosphate-containing carrier, to autologous iliac crest bone graft (ICBG) in a lumbar interbody fusion model. SUMMARY OF BACKGROUND DATA: Treatment of lumbar disk degeneration often involves spinal fusion to reduce pain and motion at the affected spinal segment by insertion of a cage containing bone graft material. Three graft materials were compared in this study-ICBG and OsteoAdapt SP (low or high dose). METHODS: The sheep underwent lateral lumbar fusion surgery with PEEK or Titanium interbody cages packed with OsteoAdapt SP (low or high dose) or ICBG. Outcomes were evaluated at 8-, 16- and 26- weeks. Newly formed bone quality, bone mineralization, and fusion were assessed by manual palpation, qualitative and semi-quantitative histopathology, histomorphometry, computed tomography (CT), and micro-CT (mCT) analysis. RESULTS: OsteoAdapt SP was implanted into 43 animals and ICBG into 21 animals (L3-L4). No group showed evidence of systemic toxicity by multiple assessments. All levels were fused by manual palpation at 26 weeks. Serial CT scans showed increasing fusion scores over time. Both doses of OsteoAdapt SP resulted in robust new bone formation and progression of fusion in the interbody cage. Range of motion tests for treatment groups was lower compared with ICBG at 8- and 16 weeks. Similarly, histology at eight weeks demonstrated more robust new bone formation for both OsteoAdapt SP groups compared to autograft. CONCLUSION: We have demonstrated the preclinical safety and efficacy of OsteoAdapt SP in a clinically relevant large animal model, supporting faster and more robust new bone formation within the interbody cage, comparable to or better than the gold standard, ICBG, in all measures.
Assuntos
Proteína Morfogenética Óssea 2 , Transplante Ósseo , Cerâmica , Vértebras Lombares , Fusão Vertebral , Animais , Fusão Vertebral/métodos , Vértebras Lombares/cirurgia , Vértebras Lombares/diagnóstico por imagem , Ovinos , Transplante Ósseo/métodos , Osteogênese/efeitos dos fármacos , Osteogênese/fisiologia , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/administração & dosagem , Ílio/transplante , Fosfatos de Cálcio , Microtomografia por Raio-X , Benzofenonas , Polímeros , Degeneração do Disco Intervertebral/cirurgia , Fator de Crescimento Transformador betaRESUMO
Joint replacements provide pain free movement for the injured or our aging population. Current prothesis mainly consist of hard metal on metal, or ceramic femoral head on ultra-high-molecular weight polyethylene (UHMWPE). In this study, a rodent fracture model was used to test the influence of wear debris from a high-performance polymer (polyimide MP-1™). Saline, MP-1™ Low Dose in Saline (1%), or MP-1 High Dose (2%) in Saline was injected directly into a standard closed unilateral femoral fracture in 12-week old Sprague Dawley rats (n = 25) for 1, 3 and 6 weeks. Endpoints included radiography, micro-computed tomography, mechanical testing and paraffin histology. No adverse effects from the wear particles were observed from the current study based on radiology, mechanical or histological data. Although the particles were present, histological analysis revealed a progression in healing between the Polyimide treated groups and the non-treated saline control groups over the duration of 1, 3, and 6 weeks, with no inhibition from the particles. The MP-1™ wear debris generated are larger than 1 µm thus are not able to be engulfed by macrophages and cause osteolysis. This family of polymers (polyimides) may be an ideal material to consider for articulating joints and other implants in the human body.
Assuntos
Consolidação da Fratura , Prótese de Quadril , Humanos , Animais , Ratos , Idoso , Microtomografia por Raio-X , Ratos Sprague-Dawley , Polietilenos/efeitos adversos , Macrófagos , Falha de Prótese , Prótese de Quadril/efeitos adversosRESUMO
BACKGROUND: In our earlier study on the functional limits of the aneurysmal aortic root we determined the pig root is susceptible to failure at high aortic pressures levels. We established a pig rupture model using cardiopulmonary bypass to determine the most susceptible region of the aortic root under the highest pressures achievable using continuous flow, and what changes occur in these regions on a macroscopic and histological level. This information may help guide clinical management of aortic root and ascending aorta pathology. METHODS: Five pigs underwent 4D flow MRI imaging pre surgery to determine vasopressor induced wall sheer stress and flow parameters. All pigs were then placed on cardiopulmonary bypass (CPB) via median sternotomy, and maximal aortic root and ascending aorta flows were initiated until rupture or failure, to determine the most susceptible region of the aorta. The heart was explanted and analysed histologically to determine if histological changes mirror the macroscopic observations. RESULTS: The magnetic resonance imaging (MRI) aortic flow and wall sheer stress (WSS) increased significantly in all regions of the aorta, and the median maximal pressures obtained during cardiopulmonary bypass was 497 mmHg and median maximal flows was 3.96 L/m. The area of failure in all experiments was the non-coronary cusp of the aortic valve. Collagen and elastin composition (%) was greatest in the proximal regions of the aorta. Collagen I and III showed greatest content in the inner aortic root and ascending aorta regions. CONCLUSIONS: This unique porcine model shows that the aortic root is most susceptible to failure at high continuous aortic pressures, supported histologically by different changes in collagen content and subtypes in the aortic root. With further analysis, this information could guide management of the aortic root in disease.
Assuntos
Aneurisma da Aorta Torácica , Ruptura Aórtica , Animais , Aorta/diagnóstico por imagem , Aorta/cirurgia , Ruptura Aórtica/diagnóstico por imagem , Valva Aórtica/diagnóstico por imagem , Valva Aórtica/cirurgia , Ponte Cardiopulmonar , SuínosRESUMO
Gene therapy continues to be a promising contender for the treatment of cystic fibrosis (CF) airway disease. We have previously demonstrated that airway conditioning with lysophosphatidylcholine (LPC) followed by delivery of a HIV-1-based lentiviral (LV) vector functionally corrects the CF transmembrane conductance regulator (CFTR) defect in the nasal airways of CF mice. In our earlier pilot study we showed that our technique can transduce marmoset lungs acutely; this study extends that work to examine gene expression in this nonhuman primate (NHP) 1 month after gene vector treatment. A mixture of three separate HIV-1 vesicular stomatitis virus G (VSV-G)-pseudotyped LV vectors containing the luciferase (Luc), LacZ, and hCFTR transgenes was delivered into the trachea through a miniature bronchoscope. We examined whether a single-dose delivery of LV vector after LPC conditioning could increase levels of transgene expression in the trachea and lungs compared with control (phosphate-buffered saline [PBS]) conditioning. At 1 month, bioluminescence was detected in vivo in the trachea of three of the six animals within the PBS control group, compared with five of the six LPC-treated animals. When examined ex vivo there was weak evidence that LPC improves tracheal Luc expression levels. In the lungs, bioluminescence was detected in vivo in four of the six PBS-treated animals, compared with five of the six LPC-treated animals; however, bioluminescence was present in all lungs when imaged ex vivo. LacZ expression was predominantly observed in the alveolar regions of the lung. hCFTR was detected by qPCR in the lungs of five animals. Basal cells were successfully isolated and expanded from marmoset tracheas, but no LacZ-positive colonies were detected. There was no evidence of an inflammatory response toward the LV vector at 1 month postdelivery, with cytokines remaining at baseline levels. In conclusion, we found weak evidence that LPC conditioning improved gene transduction in the trachea, but not in the marmoset lungs. We also highlight some of the challenges associated with translational lung gene therapy studies in NHPs.
Assuntos
Fibrose Cística , Animais , Callithrix , Fibrose Cística/genética , Fibrose Cística/terapia , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Genes Reporter , Terapia Genética , Vetores Genéticos/genética , Humanos , Lentivirus/genética , Pulmão , Camundongos , Projetos Piloto , Transdução GenéticaRESUMO
The study aimed to evaluate the comparative osteoconductivity of three commercially available bone void fillers containing gentamicin with respect to new bone, growth, host tissue response and resorption of the implant material. Defects were created in the cancellous bone of the distal femur and proximal tibia of 12-skeletally mature sheep and filled with three commercially available bone void fillers containing gentamicin (Stimulan-G, Cerament-G, Herafill-G). Peripheral blood was taken pre-operatively and at the time of implantation, as well as at intermittent timepoints following surgery to determine systemic gentamicin levels (5-,15- and 30- minutes, 1, 2, 3, 6, 12, 24, 48- and 72-hours, 3-, 6- and 12-weeks). Decalcified, embedded samples were stained with haematoxylin and eosin (H&E) and used to assess the host tissue response and the formation of new bone in the presence of test implant materials. No adverse reactions were noted at harvest at any time points for any cancellous implantation sites with the various implant materials. Comparative microCT analysis of the Stimulan-G, Cerament-G and Herafill-G test materials revealed a similar increase in bone surface area and volume between animals implanted with Stimulan-G or Cerament-G test materials. Animals implanted with Herafill-G test materials demonstrated the lowest increases in bone volume and surface area of the test materials tested, at levels similar to the negative control sites. By 12-weeks, Stimulan-G defects were completely closed with mature bone and bone marrow whilst the Cerament-G material was still present after 12 weeks by histological examination. In conclusion, this study demonstrated differences in the bone regenerative capacity of a range of bone void fillers in an in vivo setting.
Assuntos
Antibacterianos/farmacologia , Regeneração Óssea/fisiologia , Substitutos Ósseos/química , Osso e Ossos/efeitos dos fármacos , Fêmur/fisiologia , Tíbia/fisiologia , Animais , Materiais Biocompatíveis , Reabsorção Óssea , Osso e Ossos/patologia , Sulfato de Cálcio/química , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Durapatita/química , Gentamicinas/farmacologia , Teste de Materiais , Regeneração , Ovinos , Microtomografia por Raio-XRESUMO
STUDY DESIGN: This study was a multi-endpoint analysis of bone graft substitutes implanted as a standalone graft in a clinically relevant Ovine model of instrumented posterolateral spinal fusion (PLF). OBJECTIVE: The objective of this study was to obtain high-quality evidence on the efficacy of commercial bone graft substitutes compared with autograft in instrumented PLF using a state-of-the-art model with a complete range of assessment techniques. SUMMARY OF BACKGROUND DATA: Preclinical and clinical data on the quality of spinal fusions obtained with bone graft substitutes are often limited. Calcium phosphates with submicron topography have shown promising results in PLF, as these are able to induce bone formation in tissues distant from the host bone, which facilitates bony union. METHODS: Nine female, skeletally mature sheep (4-5 y) underwent posterior pedicle screw/rods instrumented PLF at L2-L3 and L4-L5 using the following bone graft materials as a standalone graft per spinal segment: (1) biphasic calcium phosphate with submicron topography (BCP<µm), (2) 45S5 Bioglass (BG), and (3) collagen-ß-tricalcium phosphate with a 45S5 Bioglass adjunct (TCP/BG). Autograft bone (AB) was used as a positive control treatment. Twelve weeks after implantation, the spinal segments were evaluated by fusion assessment (manual palpation, x-ray, micro-computed tomography, and histology), fusion mass volume quantification (micro-computed tomography), range of motion (ROM) testing, histologic evaluation, and histomorphometry. RESULTS: Fusion assessment revealed equivalence between AB and BCP<µm by all fusion assessment methods, whereas BG and TCP/BG led to significantly inferior results. Fusion mass volume was highest for BCP<µm, followed by AB, BG, and TCP/BG. ROM testing determined equivalence for spinal levels treated with AB and BCP<µm, while BG and TCP/BG exhibited higher ROM. Histologic evaluation revealed substantial bone formation in the intertransverse regions for AB and BCP<µm, whereas BG and TCP/BG grafts contained fibrous tissue and minimal bone formation. Histologic observations were supported by the histomorphometry data. CONCLUSIONS: This study reveals clear differences in efficacy between commercially available bone graft substitutes, emphasizing the importance of clinically relevant animal models with multiendpoint analyses for the evaluation of bone graft materials. The results corroborate the efficacy of calcium phosphate with submicron topography, as this was the only material that showed equivalent performance to autograft in achieving spinal fusion.
Assuntos
Substitutos Ósseos/farmacologia , Transplante Ósseo/métodos , Fosfatos de Cálcio/química , Vértebras Lombares/cirurgia , Silicatos/química , Fusão Vertebral/métodos , Animais , Fenômenos Biomecânicos , Osso e Ossos , Fosfatos de Cálcio/farmacologia , Cerâmica , Feminino , Vidro , Osteogênese/efeitos dos fármacos , Parafusos Pediculares , Amplitude de Movimento Articular , Ovinos , Microtomografia por Raio-XRESUMO
Global gene delivery to the CNS has therapeutic importance for the treatment of neurological disorders that affect the entire CNS. Due to direct contact with the CNS, cerebrospinal fluid (CSF) is an attractive route for CNS gene delivery. A safe and effective route to achieve global gene distribution in the CNS is needed, and administration of genes through the cisterna magna (CM) via a suboccipital puncture results in broad distribution in the brain and spinal cord. However, translation of this technique to clinical practice is challenging due to the risk of serious and potentially fatal complications in patients. Herein, we report development of a gene therapy delivery method to the CM through adaptation of an intravascular microcatheter, which can be safely navigated intrathecally under fluoroscopic guidance. We examined the safety, reproducibility, and distribution/transduction of this method in sheep using a self-complementary adeno-associated virus 9 (scAAV9)-GFP vector. This technique was used to treat two Tay-Sachs disease patients (30 months old and 7 months old) with AAV gene therapy. No adverse effects were observed during infusion or post-treatment. This delivery technique is a safe and minimally invasive alternative to direct infusion into the CM, achieving broad distribution of AAV gene transfer to the CNS.
Assuntos
Cisterna Magna/metabolismo , Dependovirus/genética , Expressão Gênica , Técnicas de Transferência de Genes , Vetores Genéticos/genética , Transdução Genética , Animais , Catéteres , Sistema Nervoso Central/metabolismo , Genes Reporter , Terapia Genética , Vetores Genéticos/administração & dosagem , Humanos , Injeções Espinhais , Imageamento por Ressonância Magnética , Modelos Animais , Ovinos , Cirurgia Assistida por Computador , Tomografia Computadorizada por Raios X , Transgenes , Gravação em VídeoRESUMO
Posterolateral spinal fusion (PLF) is a common procedure in orthopedic surgery that is performed to fuse adjacent vertebrae to reduce symptoms related to spinal conditions. In the current study, a novel synthetic calcium phosphate with submicron surface topography was evaluated as an autograft extender in a validated rabbit model of PLF. Fifty-nine skeletally mature New Zealand white rabbits were divided into three groups and underwent single-level intertransverse process PLF at L4-5 using (1) autologous bone graft (ABG) alone or in a 1:1 combination with (2) calcium phosphate granules (ABG/BCPgranules ), or (3) granules embedded in a fast-resorbing polymeric carrier (ABG/BCPputty ). After 6, 9, and 12 weeks, animals were sacrificed and spinal fusion was assessed by manual palpation, Radiographs, micro-CT, mechanical testing (12 weeks only), histology, and histomorphometry. Based on all endpoints, all groups showed a gradual progression in bone formation and maturation during time, leading to solid fusion masses between the transverse processes after 12 weeks. Fusion assessments by manual palpation, radiography and histology were consistent and demonstrated equivalent fusion rates between groups, with high bilateral fusion rates after 12 weeks. Mechanical tests after 12 weeks indicated substantially lower range of motion for all groups, compared to non-operated controls. By histology and histomorphometry, the gradual formation and maturation of bone in the fusion mass was confirmed for each graft type. With these results, we describe the equivalent performance between autograft and a novel calcium phosphate material as an autograft extender in a rabbit model of PLF using an extensive range of evaluation techniques. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B: 2080-2090, 2019.
Assuntos
Substitutos Ósseos , Transplante Ósseo , Fosfatos de Cálcio , Fusão Vertebral , Animais , Autoenxertos , Substitutos Ósseos/química , Substitutos Ósseos/farmacologia , Fosfatos de Cálcio/química , Fosfatos de Cálcio/farmacologia , Osteogênese , Coelhos , Propriedades de SuperfícieRESUMO
BACKGROUND CONTEXT: Degenerative disc disease (DDD) is a common, widespread socioeconomic problem. Appropriate large animal models of DDD are required for improved understanding and to serve as preclinical test beds for therapeutic strategies. PURPOSE: To evaluate the effects of short and medium duration immobilization on the sheep lumbar intervertebral disc (IVD) and facet joints (FJs), and to establish a large animal model for DDD research. STUDY DESIGN: An in vivo sheep model evaluating the effect of short- and medium-term immobilization on disc degeneration. METHODS: Eighteen sheep were equally randomized into three groups: short-term (6-week) immobilization (n=6), medium-term (26-week) immobilization (n=6), and control (no surgery) (n=6). Immobilization of L3-L4 was achieved with pedicle screw and rod implantation, the IVD was kept intact, and the annulus and end plates were not disrupted. The IVD and FJs were assessed with planar radiography, computerized tomography (CT), magnetic resonance imaging (MRI), pure moment biomechanical testing, and histologic analysis. RESULTS: Disc height was reduced for 6- and 26-week immobilization groups. The MRI and histologic analysis demonstrated significant disc degeneration for both immobilized groups compared with control, but no statistical difference was detected between short- and medium-term duration. Progressive degenerative changes in FJs were observed with micro-CT and histologic end points. Immobilization significantly reduced lateral bending and flexion-extension range of motion. CONCLUSIONS: The mechanical environment set up by immobilization alone is capable of inducing lumbar disc degeneration at both 6 and 26 weeks in sheep. Longer duration immobilization did not advance disc degeneration process beyond of that found with short duration. The present model produces a degenerative disc with intact annulus and without acute injury, more closely representing the scenario common in human disc degeneration. This provides a suitable large animal in vivo model for the evaluation of the new therapies for disc degeneration. Further studies would do well to examine the effect of remobilization after immobilization in this model.
Assuntos
Degeneração do Disco Intervertebral/veterinária , Disco Intervertebral/fisiopatologia , Vértebras Lombares/fisiopatologia , Animais , Fenômenos Biomecânicos , Modelos Animais de Doenças , Feminino , Imobilização/efeitos adversos , Disco Intervertebral/cirurgia , Degeneração do Disco Intervertebral/fisiopatologia , Vértebras Lombares/cirurgia , Imageamento por Ressonância Magnética , Parafusos Pediculares/efeitos adversos , Amplitude de Movimento Articular , Ovinos , Tomografia Computadorizada por Raios X , Articulação Zigapofisária/fisiopatologia , Articulação Zigapofisária/cirurgiaRESUMO
Management of dead space (DS) is a fundamental aspect of surgery. Residual DS following surgery can fill with hematoma and provide an environment for bacterial growth, increasing the incidence of postoperative infection. Materials for managing DS include polymethyl-methacrylate (PMMA), which is nonresorbing and requires removal in a second surgical procedure. The use of calcium sulfate (CS) offers the advantage of being fully absorbed and does not require subsequent surgical removal. As CS has historically been used as a bone void filler, there are some concerns for the risk of heterotopic ossification (HO) when implanted adjacent to soft tissue. This study assessed the osteoinductive potential of CS and identified and characterised residual material present in muscle tissue using histology, energy-dispersive X-ray spectroscopy analysis, and scanning electron microscopy (SEM). CS beads with and without antibiotic were implanted in intramuscular sites in both athymic rats and New Zealand white rabbits. At 28 days after implantation in the rat model, no signs of osteoinduction were observed. In the rabbit model, at 21 days after implantation, almost complete bead absorption and presence of a "halo" of material in the surrounding muscle tissue were confirmed. Our results suggested that the halo of material was a calcium phosphate precipitate, not HO.
Assuntos
Sulfato de Cálcio/farmacologia , Sistema Musculoesquelético/efeitos dos fármacos , Animais , Substitutos Ósseos/química , Substitutos Ósseos/farmacologia , Masculino , Teste de Materiais/métodos , Microscopia Eletrônica de Varredura/métodos , Ossificação Heterotópica/tratamento farmacológico , Polimetil Metacrilato , Próteses e Implantes , Coelhos , Ratos , Ratos NusRESUMO
BACKGROUND CONTEXT: Increasing bone ongrowth and ingrowth of polyether ether ketone (PEEK) interbody fusion devices has the potential to improve clinical outcomes. PURPOSE: This study evaluated the in vivo response of promoting new bone growth and bone apposition with NanoMetalene (NM) compared with PEEK alone in a cancellous implantation site with an empty aperture. STUDY DESIGN: This is a randomized control animal study. METHODS: Implants and funding for this study were provided by SeaSpine (60,000 USD). Cylindrical dowels with two apertures were prepared as PEEK with a sub-micron layer of the titanium (NM). The titanium coating was applied over the entire implant (Group 1) or just the apertures (Group 2). Polyether ether ketone implants with no coating served as controls (Group 3). Implants were placed in the cancellous bone of the distal femur or proximal tibia with no graft material placed in the apertures in eight adult sheep. Bone ongrowth to the surface of the implant and ingrowth into the apertures was assessed at 4 and 8 weeks after surgery with micro-computed tomography (CT) and undecalcified histology. RESULTS: The apertures in the implants were notably empty in the PEEK group at 4 and 8 weeks. In contrast, new bone formation into the apertures was found in samples coated with NM even though no graft material was placed into the defect. The bone growing into the aperture tracked along the titanium layer. Apertures with the titanium coating demonstrated significantly more bone by micro-CT qualitative grading compared with PEEK with average bone coverage scores of Group 1 (NM) 1.62±0.89, Group 2 (NM apertures only) 1.62±0.77, and Group 3 (PEEK) 0.43±0.51, respectively, at 4 weeks (p<.01) and Group 1 (NM) 1.79±1.19, Group 2 (NM apertures only) 1.98±1.18, and Group 3 (PEEK) 0.69±0.87, respectively, at 8 weeks (p<.05). The amount of bone in the apertures (ingrowth) quantified using the volumetric data from the micro-CT supported an overall increase in bone volume inside the apertures with the titanium coating compared with PEEK. Histology showed newly formed woven bone tracked along the surface of the titanium in the apertures. The PEEK interface presented the typical nonreactive fibrous tissue inside the apertures at 4 weeks and some focal contact with bone on the outside at 4 weeks and 8 weeks. CONCLUSIONS: Micro-CT and histology demonstrated bone ongrowth to the surfaces coated with NM where the newly formed bone tracked along the thin titanium-coated surfaces. Polyether ether ketone surfaces presented the nonreactive fibrous tissue at the interface as previously reported in preclinical scenarios.
Assuntos
Cetonas , Osseointegração , Polietilenoglicóis , Próteses e Implantes , Desenho de Prótese , Titânio , Animais , Benzofenonas , Osso e Ossos/fisiologia , Osso e Ossos/cirurgia , Cetonas/farmacologia , Osseointegração/efeitos dos fármacos , Osseointegração/fisiologia , Polietilenoglicóis/farmacologia , Polímeros , Próteses e Implantes/veterinária , Desenho de Prótese/métodos , Desenho de Prótese/veterinária , Distribuição Aleatória , Ovinos , Titânio/farmacologia , Microtomografia por Raio-X/métodosRESUMO
The need for bone graft materials to fill bony voids or gaps that are not related to the intrinsic stability of the bone that arise due to trauma, tumors or osteolysis remains a clinically relevant and significant issue. The in vivo response of collagen-tricalcium phosphate bone graft substitutes was evaluated in a critical size cancellous defect model in skeletally mature rabbits. While the materials were chemically virtually identical, new bone formation, implant resorption and local in vivo responses were significantly different. Differences in the in vivo response may be due, in part, collagen source and processing which influences resorption profiles. Continued improvements in processing and manufacturing techniques of collagen-tricalcium phosphate bone graft substitutes can result in osteoconductive materials that support healing of critical size bone defects even in challenging pre-clinical models.
Assuntos
Transplante Ósseo , Fosfatos de Cálcio/química , Colágeno/química , Consolidação da Fratura , Fraturas Ósseas , Animais , Regeneração Óssea , Reabsorção Óssea , Substitutos Ósseos , Feminino , Fêmur/diagnóstico por imagem , Inflamação , Microscopia Eletrônica de Varredura , Coelhos , Alicerces Teciduais , Microtomografia por Raio-XRESUMO
Decortication of bone with a high-speed burr in the absence of coolant may lead to local thermal necrosis and decreased healing ability, which may negatively impact clinical outcome. Little data are available on the impact of applying a coolant during the burring process. This study aims to establish an in vitro model to quantitatively assess peak temperatures during endplate preparation with a high-speed burr. Six sheep cervical vertebrae were dissected and mounted. Both end plates were used to give a total of 12 sites. Two thermocouples were inserted into each vertebra, 2 mm below the end plate surface and a thermal camera set up to measure surface temperature. A 3 mm high-pneumatic speed burr (Midas Rex, Medtronic, Fort Worth, TX, USA) was used to decorticate the bone in a side to side sweeping pattern, using a matchstick burr (M-8/9MH30) with light pressure. This procedure was repeated while dripping saline onto the burr and bone. Data were compared between groups using a Student's t-test. Application of coolant at the bone-burr interface during decortication resulted in a significant decrease in final temperature. Without coolant, maximum temperatures 2 mm from the surface were not sufficient to cause thermal osteonecrosis, although peak surface temperatures would cause local damage. The use of a high-speed burr provides a quick and an effective method of vertebral end plate preparation. Thermal damage to the bone can be minimized through the use of light pressure and saline coolant. This has implications for any bone preparation performed with a high-speed burr.
RESUMO
Following extensive surgical debridement in the treatment of infection, a "dead space" can result following surgical closure that can fill with hematoma, an environment conducive to bacterial growth. The eradication of dead space is essential in order to prevent recurrent infection. This study describes a novel small animal model to investigate dead-space management in muscle tissue. Two absorbable test materials were implanted in each animal; beads of calcium sulfate alone, and beads loaded with vancomycin and tobramycin. In-life blood samples and radiographs were taken from each animal following implantation. Animals were sacrificed at 1, 7, 21, 42, and 63 days post-operatively (n = 4), and implant sites were analysed by micro-computed tomography, histology and immunohistochemistry. Complete resorption was confirmed radiographically at 3 weeks post-implantation. Histologically, the host tissue response to both materials was identical, and subsequent healing at the implant sites was observed with no dead space remaining. Vancomycin was not detected in blood serum. However, peak tobramycin levels were detected in all animals at 6 hours post-implantation with no detectable levels in any animals at 72 hours post implantation. Serological inflammatory cytokine expression for IL-6, TNF-α and IL-1ß indicated no unusual inflammatory response to the implanted materials or surgical procedure. The model was found to be convenient and effective for the assessment of implant materials for management of dead space in muscle tissue. The two materials tested were effective in resolving the surgically created dead space, and did not elicit any unexpected adverse host response.
Assuntos
Implantes Absorvíveis/microbiologia , Infecções dos Tecidos Moles/microbiologia , Infecções dos Tecidos Moles/cirurgia , Implantes Absorvíveis/efeitos adversos , Animais , Desbridamento/efeitos adversos , Humanos , Interleucina-6/sangue , Modelos Animais , Coelhos , Infecções dos Tecidos Moles/sangue , Infecções dos Tecidos Moles/patologia , Fator de Necrose Tumoral alfa/sangue , Vancomicina/administração & dosagem , Cicatrização , Microtomografia por Raio-XRESUMO
Fentanyl delivered via a transdermal patch has the potential to decrease the need for post-operative handling of sheep undergoing surgical procedures. Two studies were performed to test: (1) the ideal timing for the application of pre-emptive analgesic patches and (2) the efficacy of a 2 µg/kg/h dose, as extrapolated from other species. The first study had sheep divided into two groups. Group 1 had a fentanyl patch applied for 24 h prior to a patch change and group 2 had a fentanyl patch applied 72 h prior to a change. The second study applied the results obtained in the first and tested the efficacy of 2 µg/kg/h as an effective dose in an orthopaedic surgical environment. Results indicated that the ideal time for pre-emptive fentanyl patch administration is 24-36 h prior to surgery and that 2 µg/kg/h is an effective minimum therapeutic dose rate for the use of fentanyl as an analgesic in an orthopaedic surgical environment.
Assuntos
Analgesia/veterinária , Analgésicos Opioides/farmacologia , Fentanila/farmacologia , Ovinos/cirurgia , Adesivo Transdérmico/veterinária , Analgésicos Opioides/administração & dosagem , Animais , Relação Dose-Resposta a Droga , Fentanila/administração & dosagemRESUMO
The healing of critical sized segmental defects is an ongoing clinical problem. No method has achieved pre-eminence. The Masquelet technique is a relatively new innovation involving the induction of a fibrous tissue membrane around the bone defect site taking advantage of the body's foreign body reaction to the presence of a polymethylmethacrylate (PMMA) spacer. The aim of this study was to investigate the properties and characteristics of this induced membrane and its effectiveness when used in conjunction with allograft or an allograft/autograft mix as filler materials in an ovine critical sized defect model. The resultant induced membrane was found to be effective in containing the graft materials in situ. It was demonstrated to be an organised pseudosynovial membrane which expressed bone morphogenic protein 2 (BMP2), transforming growth factor-beta (TGFß), vascular endothelial growth factor (VEGF), von Willerbrand factor (vWF), interleukin 6 (IL-6) and interleukin 8 (IL-8). While more new bone growth was evident in the test groups compared to the controls animals at 12 weeks, the volumes were not statistically different and no defects were fully bridged. Of the two graft material groups, the allograft/autograft mix was shown to have a more rapid graft resorption rate than the allograft only group. While the Masquelet technique proved effective in producing a membrane to enclose graft materials, its ability to assist in the healing of critical sized segmental defects when compared to empty controls remained inconclusive.
Assuntos
Osso e Ossos/patologia , Cicatrização , Animais , Proteína Morfogenética Óssea 2/metabolismo , Substitutos Ósseos , Osso e Ossos/metabolismo , Polimetil Metacrilato , Ovinos , Fator de Crescimento Transformador beta/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
A segmental tibial defect model in a large animal can provide a basis for testing materials and techniques for use in nonunions and severe trauma. This study reports the rationale behind establishing such a model and its design and conclusions. After ethics approval of the study, aged ewes (older than 5 y; n = 12) were enrolled. A 5-cm mid diaphyseal osteoperiosteal defect was made in the left tibia and was stabilized by using an 8-mm stainless-steel cross-locked intramedullary nail. Sheep were euthanized at 12 wk after surgery and evaluated by using radiography, microCT, and soft-tissue histology techniques. Radiology confirmed a lack of hard tissue callus bridging across the defect. Volumetric analysis based on microCT showed bone growth across the 16.5 cm(3) defect of 1.82 ± 0.94 cm(3). Histologic sections of the bridging tissues revealed callus originating from both the periosteal and endosteal surfaces, with fibrous tissue completing the bridging in all instances. Immunohistochemistry was used to evaluate the quality of the healing response. Clinical, radiographic, and histologic union was not achieved by 12 wk. This model may be effective for the investigation of surgical techniques and healing adjuncts for nonunion cases, where severe traumatic injury has led to significant bone loss.
Assuntos
Pinos Ortopédicos , Fixação Interna de Fraturas/métodos , Consolidação da Fratura/fisiologia , Modelos Animais , Fraturas da Tíbia/patologia , Animais , Calo Ósseo/crescimento & desenvolvimento , Feminino , Técnicas Histológicas , Imuno-Histoquímica , Masculino , Ovinos , Microtomografia por Raio-XRESUMO
Appropriate well-characterized bone defect animal models remain essential for preclinical research. This pilot study demonstrates a relevant animal model for cancellous bone defect healing. Three different defect diameters (8, 11, 14 mm) of fixed depth (25 mm) were compared in both skeletally immature (18-month-old) and aged sheep (5-year-old). In each animal, four defects were surgically created and placed in the cancellous bone of the medial distal femoral and proximal tibial epiphyses bilaterally. Animals were euthanized at 4 weeks post-operatively to assess early healing and any biological response. Defect sites were graded radiographically, and new bone formation quantified using µCT and histomorphometry. Fibrous tissue was found within the central region in most of the defects with woven bone normally forming near the periphery of the defect. Bone volume fraction [bone volume (BV)/TV] significantly decreased with an increasing defect diameter. Actual BV, however, increased with defect diameter. Bone ingrowth was lower for all defect diameters in the aged group. This pilot study proposes that the surgical creation of 11 mm diameter defects in the proximal tibial and distal femoral epiphyses of aged sheep is a suitable large animal model to study early healing of cancellous bone defects. The refined model allows for the placement of four separate bone defects per animal and encourages a reduction in animal numbers required for preclinical research.