Rabbit Cranial Defect Research Articles

Development of a guided bone regeneration device using salicylic acid‐poly(anhydride‐ester) polymers and osteoconductive scaffolds

Successful repair of craniofacial and periodontal tissue defects ideally involves a combined therapy that includes inflammation modulation, control of soft tissue infiltration, and bone regeneration. In this study, an anti-inflammatory polymer, salicylic acid-based poly(anhydride-ester) (SAPAE) and a three-dimensional osteoconductive ceramic scaffold were evaluated as a combined guided bone regeneration (GBR) system for concurrent control of inflammation, soft tissue ingrowth, and bone repair in a rabbit cranial defect model. At time periods of 1, 3, and 8 weeks, five groups were compared: (1) scaffolds with a solid ceramic cap (as a GBR structure); (2) scaffolds with no cap; (3) scaffolds with a poly(lactide-glycolide) cap; (4) scaffolds with a slow release SAPAE polymer cap; and (5) scaffolds with a fast release SAPAE polymer cap. Cellular infiltration and bone formation in these scaffolds were evaluated to assess inflammation and bone repair capacity of the test groups. The SAPAE polymers suppressed inflammation and displayed no deleterious effect on bone formation. Additional work is warranted to optimize the anti-inflammatory action of the SAPAE, GBR suppression of soft tissue infiltration, and stimulation of bone formation in the scaffolds and create a composite device for successful repair of craniofacial and periodontal tissue defects.

Negative pressure technology enhances bone regeneration in rabbit skull defects

BackgroundBone is a slowly regenerating tissue influenced by various physiological processes, including proliferation, differentiation, and angiogenesis, under the control of growth factors. Shortening this healing time is an important and popular clinical research focus in orthopedics. Negative pressure can stimulate angiogenesis, improve blood circulation, promote granulation tissue growth and accelerate tissue wound healing. We sought to determine whether negative pressure could reduce bone healing time in a rabbit cranial defect model.MethodsFour symmetrical holes (diameter, 3.5 mm) were drilled into the skulls of 42 New Zealand white rabbits, with two holes in each parietal bone. For each rabbit, the two sides were then randomly assigned into experimental and control groups. Using negative pressure suction tubes, experimental holes were treated with −50 kPa for 15 minutes, four times per day, whereas the control holes remained untreated. After 4 weeks, the negative pressure suction tubes were removed. At 2, 4, 6 and 8 weeks, three-dimensional (3D) reconstruction computed tomography (CT), X-ray radiopacity, and two-photon absorptiometry were used to evaluate new bone formation. Histological changes were determined by hematoxylin and eosin (H.E) staining. At weekly intervals until 6 weeks, the mRNA expression levels of vascular endothelial growth factor (VEGF) and bone morphogenetic protein (BMP)-2 were evaluated by RT-PCR. A paired student’s t-test was employed to compare X-ray radiopacity and bone density measurements between the experimental and control groups.Results3D-reconstruction CT showed that new bone regeneration in the experimental group was greater than that in the control group at 4 and 6 weeks. At these time points, the experimental group presented with higher X-ray radiopacity and increased bone density (P < 0.05) as compared with the control group. Cartilage islands and new bone were observed by H.E staining at 2 weeks in the experimental group. By 6 weeks, the new bone had matured into lamellar bone in the experimental group. RT-PCR results showed that VEGF and BMP-2 were highly expressed in the experimental group as compared with control.ConclusionsIntermittent negative pressure can promote the regeneration of bone possibly by enhancing the expression of VEGF and BMP-2.

Biological activity of a nanofibrous barrier membrane containing bone morphogenetic protein formed by core‐shell electrospinning as a sustained delivery vehicle

This study reports the in vitro and in vivo biological activities of recombinant human bone morphogenetic protein 2 (rhBMP-2) released from the core-shell structure of a nanofibrous barrier membrane as a sustained delivery model for bone regeneration. RhBMP-2 incorporating poly(ethylene glycol) was used as the core, and poly(caprolactone) was used as the shell surrounding the core. To determine its release profiles, the release solution was collected and the amount of rhBMP-2 was measured by ELlSA at different time points. In vitro rhBMP-2, released from the delivery system over at least 24 days, reached a stable rate of 500 pg per day and guided bone marrow mesenchymal stem cells (BMMSCs) to express osteogenic genes. The distribution and proliferation of BMMSCs in the nanofibrous barrier membrane was measured by laser confocal scanning microscopy (LCSM) and scanning electron microscopy (SEM). The biological activity of rhBMP-2 was tested in BMMSC/membrane culture in vitro and in a rabbit calvarium defect model in vivo. Osteogenic genes osteonectin (ON) and core binding factor-α1 (Cbf-α1) expression of the BMMSCs cultured on the BMP-2-PEG/PCL membrane were significantly higher than those of cells on the PEG/PCL membrane at the late time points using real-time PCR (p < 0.05). The membranes containing the rhBMP-2 group exhibited the fastest and most bone formation compared to others in rabbit cranial defect models (p < 0.05). This study revealed that rhBMP-2 could be incorporated into a core-shell electrospun membrane, and preserve sustained release capability and biological activity.

Cranial Particulate Bone Graft Ossifies Calvarial Defects by Osteogenesis

Cranial particulate bone graft heals inlay calvarial defects and can be harvested as early as infancy. The purpose of this study was to test the hypothesis that particulate bone promotes ossification primarily by osteogenesis. Freshly harvested particulate bone, devitalized particulate bone, and high-speed drilled bone dust from rabbit calvaria were assayed for metabolic activity (resazurin) and viable osteoblasts (alkaline phosphatase). A rabbit cranial defect model was used to test the effect of devitalizing particulate bone on in vivo ossification. A parietal critical-size defect was created and managed in three ways: (1) no implant (n = 6); (2) particulate bone implant (n = 6); and (3) devitalized particulate bone implant (n = 6). Micro-computed tomographic scanning was used to measure ossification 16 weeks later; histology also was studied. Particulate bone contained more viable cells (0.94 percent transmittance per milligram) compared with devitalized particulate bone (0.007 percent) or bone dust (0.21 percent) (p = 0.01). Particulate bone had greater alkaline phosphatase activity (0.13 μU/μg) than devitalized particulate bone (0.000) or bone dust (0.06) (p = 0.01). Critical-size defects treated with particulate bone had more ossification (99.7 percent) compared with devitalized particulate bone implants (42.2 percent) (p = 0.01); no difference was found between devitalized particulate bone and the control (40.8 percent) (p = 0.9). Particulate bone graft contains living cells, including osteoblasts, that are required to heal critical-size cranial defects. These data support the hypothesis that particulate bone promotes ossification primarily by osteogenesis.

Fabrication and in vivo evaluation of Ti6Al4V implants with controlled porous structure and complex shape

Electron beam melting process was used to fabricate porous Ti6Al4V implants. The porous structure and surface topography of the implants were characterized by scanning electron microscopy (SEM) and digital microscopy (DM). The results showed that the pore size was around 600 and the porosity approximated to 57%. There was about ±50 μm of undulation on implants surfaces. Standard implants and a custom implant coupled with porous sections were designed and fabricated to validate the versatility of the electron beam melting (EBM) technique. After coated with bone-like apatite, samples with fully porous structures were implanted into cranial defects in rabbits to investigate the in vivo performance. The animals were sacrificed at 8 and 12 weeks after implantation. Bone ingrowth into porous structure was examined by histological analysis. The histological sections indicated that a large amount of new bone formation was observed in porous structure. The newly formed bone grew from the calvarial margins toward the center of the bone defect and was in close contact with implant surfaces. The results of the study showed that the EBM produced Ti6Al4V implants with well-controlled porous structure, rough surface topography and bone-like apatite layer are beneficial for bone ingrowth and apposition.

Bioelectric effect of a chitosan bioelectret membrane on bone regeneration in rabbit cranial defects

In this study, the effects of a chitosan bioelectret membrane on bone regeneration in a rabbit cranial defect model were examined. The bioelectret was fabricated by film casting and polarized by grid-controlled corona charging (−1 kV). In vitro, the equivalence surface charge density of the bioelectret gradually reduced under both wet and dry conditions. In vivo, two rectangular transosseous defects of 8 mm × 10 mm were created symmetrically on each rabbit cranial bone. The defects were treated with the bioelectret membrane, with unpolarized membrane, and no membrane, respectively. The rabbits were killed at 4, 8, and 12 weeks postoperatively, and specimens were examined by micro-computed tomography and histological analysis. The bioelectret membrane recipients had a significantly higher volume of newly formed bone and greater rate of material degradation than the unpolarized membrane. Furthermore, the bioelectret membrane induced new bone formation not only around the host bone but also in the center of the defects. The chitosan bioelectret membrane application has an apparent potential in guided bone regeneration applications.

Evaluation of bone replacement materials in a rabbit cranial defect model using micro CT and hard tissue histology

Introduction: For functional and structural restoration of bone deficiencies, various resorbable and nonresorbable alloplastic materials have been introduced, including metals, polymers and ceramics. However, an “optimal” artificial replacement for craniofacial bone has not been found yet and the search for improved reconstruction methods and alternative materials is going on. To assess and compare biocompatibility and osseointegration of these materials, adequate animal test models are indispensable. Methods: In a rabbit cranial defect model, biocompatibility and osseointegration of polymeric and composite bone replacement materials were evaluated at different time points after implantation. Calvaria including implants and surrounding tissue were explanted and embedded in methacrylate resin. The samples were scanned with a nanotom® (phoenix|x-ray) µCT scanner and proceeded for histological examination by sawing-grinding technique. Avizo® Fire (vsg) software was used for visualisation and processing of µCT data. Qualitative and morphometric evaluation of osseointegration and fibrous encapsulation was performed on undecalcified histologic preparations of the explants, and on 3D reconstructions plus virtual slices derived from corresponding 3D µCT datasets. Results: The obtained 3D µCT data enabled a comprehensive qualitative and quantitative assessment of osseointegration and biodegradation of radioopaque composite implants. Prerequisite for visualization and discrimination of materials by µCT is a significant difference of their hounsfield values. Due to this limitation, radiolucent polymeric implant materials and soft tissue could not be distinguished from embedding resin. In contrast, histologic preparations of undecalcified hard tissue and implant materials enabled detailed visualization and examination of all tissues and implant materials. The substantial disadvantage of hard tissue histology was the inevitable loss of information due to small number of slices and large gaps between specimens yielded by this method. Conclusion: To obtain comprehensive and quantifiable information about biodegradation, biocompatibility and osseointegration of alloplastic bone replacement materials, µCT scans as well as histologic evaluation should be performed.

Comparison of the bone healing capacity of autogenous bone, demineralized freeze dried bone allograft, and collagen sponge in repairing rabbit cranial defects

Objectives: This study sought to evaluate the efficacy of collagen graft materials, as compared to other graft materials, for use in healing calvarial defects in rabbits. Materials and Methods: Ten mm diameter calvarial defects were made in ten rabbits. The rabbits were then divided into 4 groups: control, autogenous bone graft, SureOss graft, and Teruplug graft. Bone regeneration was evaluated using histological and radiographic methods. Results: Based on visual examination, no distinct healing profile was observed. At 4 weeks after treatment, histological analysis showed there was no bone regeneration in the control group; however, at 8 weeks after treatment, new bone formation was observed around the margin of the defective sites. In the autogenous bone graft group, new bone formation was observed at 4 weeks after treatment and mature bone was detected around the grafted bone after 8 weeks. In the SureOss graft group, at 4 weeks after treatment, acute inflammatory and multinuclear cells were noted around the grafted materials; at 8 weeks after treatment, a decrease in graft materials coupled with new bone formation were observed at the defective sites. In the Teruplug graft group, new bone formation was detected surrounding the bone margin and without signs of inflammation. There were statistically significant differences observed between the graft and control group in terms of bone density as evidenced by radiographic analysis using computed tomography (P Conclusion: These results suggested that autogenous bone, SureOss and Teruplug have the ability to induce bone regeneration as compared to an untreated control group. The osteogenic potential of Teruplug was observed to be lower than that of autogenous bone, but similar to that of SureOss.

Evaluation of an anorganic bovine‐derived mineral with P‐15 hydrogel bone graft: preliminary study in a rabbit cranial bone model

The present investigation aimed to assess the bone-regenerative potential of two formulations of anorganic bovine-derived mineral bound to a P-15 (ABM/P-15) bone graft - the particulate and the hydrogel forms - in a delayed healing rabbit cranial defect model. Ten adult male New Zealand White rabbits were used to create two 8 mm transcortical cranial defects per rabbit and each one received randomly the test material (ABM/P-15 carboxymethyl cellulose (CMC)-hydrogel graft), the standard control material (ABM/P-15 particulate graft) or remained empty as a negative control. The defects were allowed to heal for 2 and 4 weeks. Qualitative and quantitative histological outcomes were assessed on undecalcified sections. In the defects grafted with the test material, at both time points, there was a marked random migration of the bone substitute particles. As a consequence, the space maintenance provision was lost and new bone formation was reduced compared with the control particulate graft material. The histomorphometric analysis showed that the control material attained better results, with an average of 13.8 ± 1.9% and 18.2 ± 4.4% of new bone at 2 and 4 weeks, compared with 8.5 ± 2.4% and 13 ± 2.9% for the test material. These differences were significant at 2 weeks (P ≤ 0.05), but not at 4 weeks (P>0.05). Additionally, there was a significant difference in the total area of mineralized tissue (new bone plus particles), favoring the standard control over the test material: 43.2 ± 14.4% vs. 14.2 ± 5.3% at 2 weeks and 56.9 ± 4.2% vs. 24.2 ± 9.6% at 4 weeks, respectively. The test ABM/P-15 CMC-hydrogel graft material behaved in this animal model by migration of the graft particles, what determined an unpredictable osseoconduction and, consequently, a decreased quality and quantity of bone regeneration as compared with the osseopromotive behavior exhibited by the standard particulate form of the ABM/P-15 control graft. It is therefore suggested to restrain the application of the hydrogel graft form in non-contained anatomical bone defects.

Bone regeneration with glass ceramic implants and calcium phosphate cements in a rabbit cranial defect model

Hydroxyapatite cement (BoneSource®) and brushite calcium phosphate cement (chronOS™ Inject) were tested for fixation of glass ceramic implants (Bioverit®) in experimentally created cranial defects in 24 adult New Zealand White rabbits. Aim of the in vivo study was to assess and compare the biocompatibility and osseointegration of the implanted materials. Macroscopic and histological evaluations were performed 1 month, 3 months, and 6 months postoperatively. All implanted materials were well tolerated by the surrounding tissue. Both bone cements exhibited osteoconductive properties. Differences could be detected regarding to the rates of cement resorption and new bone formation. The brushite cement was resorbed faster than the hydroxyapatite cement. The chronOS™ Inject samples exhibited a higher rate of connective tissue formation and an insufficient osseointegration. BoneSource® was replaced by bone with minimal invasion of connective tissue. New bone formation occurred faster compared to the chronOS™ Inject group. Bioverit® implants fixed with BoneSource® were successfully osseointegrated.

Effects of Platelet-Rich Plasma and Chitosan Combination on Bone Regeneration in Experimental Rabbit Cranial Defects

This study aimed to investigate and compare the effect of chitosan sponge and platelet-rich plasma (PRP) gel alone as well as their combination on bone regeneration in rabbit cranial defects. Four cranial defects with a 4.5-mm diameter were created in rabbit cranium and grafted with PRP, chitosan sponge alone, and chitosan sponge incorporated with PRP. The rabbits were killed by the fourth and eighth weeks, and the defects were analyzed histologically. Higher bone formation was observed in the PRP group when compared with the other groups at weeks 4 and 8. All parts of the defects were filled with thick trabecular new bone in the PRP group. The amount of new bone formation in the control groups was found to be less when compared with the PRP group and the least in the chitosan group. The defects that were filled with chitosan sponge showed a limited amount of new bone formation and an obvious fibrous demarcation line between chitosan particles and bone. Application of PRP showed a histological tendency toward increased bone formation. Other forms or derivatives of chitosan may have beneficial effects to achieve new bone regeneration.

Spontaneous healing capacity of rabbit cranial defects of various sizes

PurposeThis study evaluated the spontaneous healing capacity of surgically produced cranial defects in rabbits with different healing periods in order to determine the critical size defect (CSD) of the rabbit cranium.MethodsThirty-two New Zealand white rabbits were used in this study. Defects of three sizes (6, 8, and 11 mm) were created in each of 16 randomly selected rabbits, and 15-mm defects were created individually in another 16 rabbits. The defects were analyzed using radiography, histologic analysis, and histometric analysis after the animal was sacrificed at 2, 4, 8, or 12 weeks postoperatively. Four samples were analyzed for each size of defect and each healing period.ResultsThe radiographic findings indicated that defect filling gradually increased over time and that smaller defects were covered with a greater amount of radiopaque substance. Bony islands were observed at 8 weeks at the center of the defect in both histologic sections and radiographs. Histometrical values show that it was impossible to determine the precise CSD of the rabbit cranium. However, the innate healing capacity that originates from the defect margin was found to be constant regardless of the defect size.ConclusionsThe results obtained for the spontaneous healing capacity of rabbit cranial defects over time and the underlying factors may provide useful guidelines for the development of a rabbit cranial model for in vivo investigations of new bone materials.

Correlation Between Micro-Computed Tomography and Histomorphometry for Assessment of New Bone Formation in a Calvarial Experimental Model

Conventional histologic or histomorphometric evaluation provides clear evidence of the bone healing process. However, the sample preparation process is tedious and destructive, and the three-dimensional (3D) anisotropic information of the bone trabeculae is compromised. Micro-computed tomography (microCT) has been introduced as an alternative to these traditional evaluation methods. microCT is noninvasive and provides a faster approach to evaluate and quantify cancellous bone. Most previous studies that used microCT have focused on studying trabecular structures of cancellous bone. In this study, we used microCT to analyze the micro-architecture of the regenerated membranous bone using a rabbit cranial defect model. Two 1 cm diameter circular bony defects were created in 12 New Zealand white rabbits. Specimens were harvested at 6 weeks and 12 weeks after surgery and were scanned using a MicroCT machine (Skyscan 1072, Aartselaar, Belgium). The specimens were then sectioned and stained with Goldner's trichrome. Bone volume density (BV/TV), bone surface density (BS/BV), and trabecular thickness (TbTh) were determined from histomorphometric and two-dimensional (2D) and 3D microCT analysis. Pearson's correlation coefficient (gamma), paired t-tests, and intraclass correlation coefficients from measurements between the 2D and 3D microCT and histomorphometry were calculated. There were very strong positive correlations of BV/TV between histomorphometric and 2D or 3D microCT measurements. Correlation between histomorphometric and 2D microCT measurements for BS/BV was moderate, whereas correlation between histomorphometric and 3D microCT measurements was weak. Weak correlations in TbTh among the three methods were found. In conclusion, the present study suggests that, in evaluating micro-architectures in regenerated bones, the correlation between measuring methods vary according to the features measured.

Cranial Defect Reconstruction in an Experimental Model Using Different Mixtures of Bioglass and Autologous Bone

The purpose of this study was to investigate bone healing in a rabbit cranial defect model using different mixtures of bioglass (NovaBone C/M; Porex Surgical, Inc., Newnan, GA) and autologous bone with and without an overlying absorbable plate (Lactosorb; Walter Lorenz Surgical, Jacksonville, FL). Twelve rabbits were divided into three groups, and a 2 cm diameter cranial defect was created in each rabbit. Group I defects were filled with 80% bioglass and 20% autologous bone, group II with 60% bioglass and 40% autologous bone, and group III with 60% bioglass and 40% autologous bone with the addition of an absorbable plate placed directly over the reconstructed defect to help separate it from the overlying soft tissues. Rabbits were euthanized 6 months postoperatively. Histologic examination was then performed. The size of the remaining bone gap, area of reossification within the defect, and percentage of fibrous tissue within the defect were measured. Histologic analyses revealed that group II animals had an increased cross-sectional area of bone formation and decreased bone gap when compared with group I and III animals (P < 0.05). There were no statistical differences between groups I and III with regard to bone formation. This study suggests that when combining bioactive glass and autologous bone for repairing cranial defects, a combination of 60% bioactive glass and 40% bone graft yields superior results to a combination of 80% bioactive glass and 20% bone graft. Placing an absorbable plate over a defect filled with 60% bioactive glass and 40% bone graft inhibits rather than promotes reossification.

Poster 109: The Autogenous Bone Marrow Cells Transplant Using Atherocollagen Carrier to Rabbit Cranial Bone Defect

Bone transplants are mainly performed for oral bone defects. Autogenous bone transplants are performed for large bone defects, but there are problems such as limited quantities of bone and the occurrence of postoperative infection. Therefore, a tissue engineering method has been receiving attention as an alternative method of bone transplantation, and in our department, it has been confirmed that bone formation occurs when bone-marrow cells are transplanted with AC (atherocollagen) as a carrier. However, further experiments using autogenous cell transplantation are necessary before clinical application can be carried out. So far there have only been few such studies and therefore many questions remain to be clarified. In our experiment, we performed autogenous cell transplants on rabbits using AC as a carrier. Bone-marrow cells were taken from the right thigh bones of Japanese white rabbits (male, approximately 3 kg) and then were cultured for 3 weeks in order to biochemically investigate bone neoplasia in vitro as part of Experiment 1. For Experiment 2, in parietal bone defects in rabbits, group A had a carrier attached to the cells, group B had a carrier only, and group C had bone defects only. Five rabbits from each group were killed after 2, 4, and 12 weeks to perform comparative investigations of both μ-CT and pathology. In Experiment 1, it was found that bone-marrow cells had calcium deposits and bone neoplasia was also detected. In Experiment 2, bone formation was seen in group A and group B in which the carrier was used, and group A tended to demonstrate more such bone formation than group B. Furthermore, bone formation was less frequently detected in group C, in which the carrier was not used. Our experiment revealed that attaching a carrier to the cells thus resulted in more extensive bone formation and a higher efficacy than using a carrier alone, thus indicating that this may be a potentially effective alternative method for bone transplantation in the future. In our experiment, this method of attaching an AC carrier to cells was thus shown to be highly effective. By using AC, which has already been applied in a clinical setting for autogenous cells, our experimental method was thus found to be potentially useful for application in an early clinical setting.

Heterobifunctional Poly(Ethylene Glycol)–Tethered Bone Morphogenetic Protein-2–Stimulated Bone Marrow Mesenchymal Stromal Cell Differentiation and Osteogenesis

We describe a biomimetic mode of insoluble signaling stimulation to provide target delivery of bone morphogenetic protein-2 (BMP-2), with the aim of prolonging the retention of BMP-2 use in bone tissue engineering and to enable its localized release in response to cellular activity. In our novel localization process, we used heterobifunctional acrylate-N-hydroxysuccinimide poly(ethylene glycol) (PEG) as a spacer to tether BMP-2 onto a poly(lactide-co-glycolide) scaffold. Use of PEG-tethered BMP-2 was feasible because BMP-2 retained its activity after covalent conjugation. The PEG-tethered BMP-2 conjugate sustained stimulation and retained its mitogenic activity, notably affecting pluripotent stem cell proliferation and differentiation. We seeded the scaffolds with bone marrow-derived mesenchymal stromal cells as progenitor cells to evaluate their morphology and phenotypic expression. We also created bilateral, full-thickness cranial defects in rabbits to investigate the osteogenic effect of cultured mesenchymal stromal cells on bone regeneration in vivo. Histomorphometry and histology demonstrated that the PEG-tethered BMP-2 conjugate enhanced de novo bone formation after surgery. Our work revealed the potential for biomimetic surface engineering by entrapping signaling growth factor to stimulate osteogenesis. Our technique may provide a new platform for bone-engineered stem cell therapies.

Osteopontin and bone metabolism in healing cranial defects in rabbits

Non-collagen proteins such as bone sialoprotein and osteopontin (OPN) form 10% of the extracellular bone matrix. In this study, the influence of OPN on bone repair was investigated. Human OPN (Innogenetics ®) was produced by a recombinant technique and bonded onto the surface of hydroxyapatite (Interpore 200 ®). Thirty rabbits were divided into six equal groups. A circular defect (10 mm) was prepared in each parietal bone. In four rabbits of each group the left and right defects were filled with either OPN-coated hydroxyapatite (OPN-HA) or non-coated hydroxyapatite (HA). One sham animal of each group received no implants. The animals were killed after 1, 2, 6, 12, 18 and 30 weeks. The histological sections were scanned and analysed digitally. There were no statistically significant differences in total bone formation between defects filled with OPN-HA and HA. Bone formation at the borders of the healing area was significantly higher in defects filled with OPN-HA than in those filled with HA. Less bone formation was noted in the OPN-HA and HA groups at the centre of the healing area than at the borders of the healing area and the dural area. Although some animals in the sham group showed a high level of bone formation in the dural area, this was not significantly different to that in the dural area of the other groups. There was no sign of infection or tissue rejection of the graft.

Biodegradation process of α‐TCP particles and new bone formation in a rabbit cranial defect model

The purpose of the present study was to observe the biodegradation process of pure alpha-tricalcium phosphate (alpha-TCP) particles and to determine the efficacy of alpha-TCP as a space maintainer in a bone defect. We used 14 rabbits and prepared two cranial bone defects in each rabbit. One defect was left empty as a control, whereas the other was filled with alpha-TCP particles about 300 mum in diameter. Animals were sacrificed at 1 week, 4 weeks, and 8 weeks. The cranial bone was then embedded either in paraffin wax for the preparation of decalcified specimens, or in polyester resin for the preparation of nondecalcified specimens. All specimens were evaluated histologically and histomorphometrically. As a consequence of the degradation of alpha-TCP, a "reticulate structure" appeared in the particles at 1 week and new bone was observed in this structure at 8 weeks. The amount of new bone between the control and experimental groups was not significantly different at any of the time points. However, in the experimental group, new bone at the surface of alpha-TCP was evident even in the center of the defect whereas fibrous connective tissue was dominant in the control group. These results indicate that alpha-TCP is a degradable osteoconductive material that is able to act as a space maintainer for bone regeneration when applied to a bone defect. While there was no significant difference in total bone formation between the experimental and negative control groups, the space-maintaining and osteoconductive properties of the particles may result in more complete bone formation in longer-term studies.

Early Dural Reaction to Polylactide in Cranial Defects in Rabbits

Restoring the bone integrity to injured calvariae remains a challenge to surgeons. In this study, the dural biocompatibility of biodegradable poly-L/DL-lactide 80/20 and 70/30 defect covers, designed for guided bone regeneration, was assessed. In each of the 16 test rabbits, bilateral (8.3 mm) cranial defects were created. The different covers were applied to one defect each in every rabbit and consisted of three parts: an epicranial cover, a spacer, and a dural cover. All defects had closed after 8 weeks due to new bone formation. A few giant cells were found at the cover-to-dura interface in equal numbers for both covers. Dural bone formation was present in 15 of 16 rabbits and progressed unhindered by the defect cover or its early degradation products.

Effect of Transforming Growth Factor β2on Marrow-Infused Foam Poly(Propylene Fumarate) Tissue-Engineered Constructs for the Repair of Critical-Size Cranial Defects in Rabbits

This study investigates the osseointegration of poly(propylene fumarate) (PPF) with beta-tricalcium phosphate (beta-TCP) scaffolds in a critical-size (diameter, 1.6 cm), cranial defect in 4-month-old rabbits (n = 51), killed at 6 or 12 weeks. Two molecular weights of PPF were used to produce bilayer scaffolds with 0.5-mm solid external and 2.0-mm porous internal layers. The porous layer was infused with bone marrow aspirate, with half the animals receiving 0.8 microg of transforming growth factor beta2 (TGF-beta2). No foreign body or inflammatory response was observed externally or on histological examination of explants. Statistical analysis of histological areal and linear measures of new bone formation found significantly more bone at the later sacrifice time, followed by implants receiving TGF-beta2, followed by low molecular weight PPF implants. Approximately 40% of the explants were tested for incorporation strength with a one-point "push-in" test. Because no permanent fixation was used, implant strength (28.37-129.03 N; range, 6.4 to 29.0 lb of resistance) was due entirely to new bone formation. The strongest bone was seen in implants receiving TGF-beta2-infused marrow in animals killed at 12 weeks. These results support the use of PPF as an osteogenic substrate and future research into preoperative fabrication of critical size and supercritical-size cranial prosthetic implants.