Rabbit Calvaria Defects Research Articles

Comparing the effects of Bone+B® xenograft and InterOss® xenograft bone material on rabbit calvaria bone defect regeneration.

The bone regeneration therapy is often used in patients with inadequate bone support for implants, particularly following tooth extractions. Xenografts derived from animal tissues are effective bone reconstructive options that resist resorption and pose a low risk of transmitting disease. Therefore, these implants may be a good option for enhancing and stabilizing maxillary sinuses. The purpose of this study was to compare two xenografts, Bone+B® and InterOss®, for the reconstruction of rabbit calvaria defects. The study involved seven male New Zealand white rabbits. In the surgical procedure, 21 spots were created on both sides of the midline calvarium by creating three 8-millimeter defects. A control group was used, as well as two treatment groups utilizing Bone+B® Grafts and InterOss® Grafts. After 3 months, the rabbits were euthanized, followed by pathological evaluation. Analysis of these samples focused on bone formation, xenograft remaining material, and inflammation levels, using Adobe Photoshop CS 8.0 and SPSS version 24. With the application of Bone+B® graft, bone formation ranged from 32% to 45%, with a mean of 37.80% (±5.63), and the remaining material ranged from 28% to 37%, with a mean of 32.60% (±3.65). Using InterOss® grafts, bone formation was 61% to 75%, the mean was 65.83% (±4.75), and the remaining material was 9% to 18%, with a mean of 13.17% (±3.06). The bone formation in the control group ranged from 10% to 25%, with a mean of 17.17% (±6.11). InterOss® had lower inflammation levels than other groups, but the difference was not statistically significant (p > .05). InterOss® bone powder is the best option for maxillofacial surgery and bone reconstruction. This is due to more bone formation, less remaining material, and a lower inflammation level. Compared to the control group, Bone+B® improves healing and bone quality, thus making it an alternative to InterOss®.

RETRACTION: Biphasic β-TCP mixed with silicon increases bone formation in critical site defects in rabbit calvaria.

J. L. Calvo-Guirado, M. Garces, R. A. Delgado-Ruiz, M. P. Ramirez Fernandez, E. Ferres-Amat, G. E. Romanos, "Biphasic β-TCP mixed with silicon increases bone formation in critical site defects in rabbit calvaria", Clinical Oral Implants Research 26, no. 8 (2015): 891-897, https://doi.org/10.1111/clr.12413 The above article, published online on 26 May 2014 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the journal's Editor in Chief, Lisa J. A. Heitz-Mayfield; and John Wiley & Sons Ltd. The retraction has been agreed due to concerns raised by third parties on the data presented in the article. Several flaws and inconsistencies between results presented and experimental methods described were found. Specifically, Figure2b has been used in a different context in another publication authored by the same group. Additionally, Figure3 and Figure5e appear to derive from the same histological sections depicted in another publication authored by the same group where it is used in a different context. Accordingly, the editors consider the findings of this article unreliable and the conclusions to be invalid. The authors have been informed of the decision to retract the article. The corresponding author J. L. Calvo-Guirado and co-authors M. Garces and R. A. Delgado-Ruiz disagree with the decision of retraction. The remaining co-authors were not available for a final confirmation of the retraction.

Comparison of the healing process of xenografts with three different sources in critical-size bone defects: An in vivo study.

Xenograft bone substitutes can be obtained from different animals and processed using various methods. The present in vivo study evaluated bone regeneration after using three types of xenografts with different sources in critical-sized bone defects in rabbit calvaria. Four 8-mm defects were created in calvaria of 14 New Zealand and white male rabbits. Three out of four defects were filled with xenografts of bovine, camel, and ostrich sources. The fourth defect was left unfilled as the control group. Seven rabbits were sacrificed after eight weeks and seven others after 12 weeks. Micro-CT imaging and histologic evaluation were further performed on dissected calvarias. After 8 and 12 weeks, the highest and lowest percentages of new bone formation were observed in the camel (27.71% and 41.92%) and control (11.33% and 15.96%) groups, respectively. In the case of residual material, the ostrich group had the most value after eight weeks (53%), while after 12 weeks, it was highest in the camel group (37%). Micro-CT findings were consistent with histologic results. Although all three xenografts can be good choices for treating bone defects, camel-sourced xenograft seemed to be better than the other two groups. The origin and processing procedures of xenografts affected their final characteristics, which should be considered for clinical use.

Comparison of the Volume and Histological Properties of Newly Formed Bone after the Application of Three Types of Bone Substitutes in Critical-Sized Bone Defects.

This study aimed to compare the volume and quality of the newly formed bone following application of two types of xenografts and one synthetic material in bone defects in rabbit calvaria from histological and micro-CT aspects. Four 8-mm defects were created in 12 rabbit calvaria. Three defects were filled with bone substitutes and one was left unfilled as the control group. The newly formed bone was evaluated histologically and also by micro-CT at 8 and 12 weeks after the intervention. The percentage of osteogenesis was comparable in histomor-phometric assessment and micro-CT. Histological analysis showed that the percentage of the newly formed bone was 10.92 ± 5.17%, 14.70 ± 11.02%, 11.47 ± 7.04%, and 9.45 ± 5.18% in groups bovine 1, bovine 2, synthetic, and negative control, respectively after 8 weeks. These values were 33.70 ± 11.48%, 26.30 ± 18.05%, 22.92 ± 6.30%, and 14.82 ± 8.59%, respectively at 12 weeks. The difference in the percentage of the new bone formation at 8 and 12 weeks was not significant in any group (P > 0.05) except for bovine 1 group (P < 0.05). Micro-CT confirmed new bone formation in all groups but according to the micro-CT results, the difference between the control and other groups was significant in this respect (P < 0.05). All bone substitutes enhanced new bone formation compared with the control group. Micro-CT assessment yielded more accurate and different results compared with histological assessment.

The Use of Human Sterilized Crushed Tooth Particles Compared with BTCP Biomaterial and Empty Defects in Bone Formation inside Critical Rabbit Calvaria Sites.

This study aimed to assess the bone regeneration of critical-size defects in rabbit calvaria filled with freshly crushed extracted teeth, comparing them with BTCP biomaterial and empty sites. Materials and methods: Twenty-one female New Zealand rabbits were used in this study. Two critical-size defects 6 mm in size were created in the skull bone, each with a 3 mm separation between them. Three experimental groups were evaluated: Group A (human sterilized crushed teeth granules alone), Group B (Bioner Bone, Bioner Sitemas Implantológicos), and Group C (unfilled defects). The animals were sacrificed at 4 and 8 weeks. Evaluation of the samples involved histological and histomorphometric analyses with radiographic evaluation. The histological evaluation showed a higher volume reduction in Group A compared with Group B (p < 0.05) and Control. Group A showed the highest values for cortical closure and bone formation around the particles, followed by Group B and Group C (p < 0.05). Within the limitations of this animal study, we can conclude that the use of human tooth particles leads to increased bone formation and reduced connective tissue in critical-size defects in rabbit calvaria when compared to BTCP biomaterial. The calvarial model is a robust base for the evaluation of different biomaterials.

Augmented Repair and Regeneration of Critical Size Rabbit Calvaria Defects with 3D Printed Silk Fibroin Microfibers Reinforced PCL Composite Scaffolds

Treatment of critical size defects is quite challenging, often requiring autologous bone grafts for bone regeneration. A massive volume of autologous bone is essential during this process to fill the defect leading to donor site morbidity. Although 3D printed PCL scaffolds are frequently utilised for bone correction procedures, there have been reports of delayed PCL biodegradation and inadequate bone tissue formation. To enhance the regenerative potential, in this study, silk in the form of silk fibroin microfibers are reinforced into the PCL matrix to form the composite. Two silk variations were used: Antheraea mylitta and Bombyx mori, and has been proven to promote cell proliferation, adhesion, and osteogenic potential. This work creates 8 mm critical size defects in a rabbit calvaria model to test for the first time ever the ability of 3D printed PCL-silk scaffolds to regenerate bone tissue. Micro-CT imaging and histological examination performed 6 and 12 weeks after implantation revealed that the PCL-silk scaffold-augmented defects considerably outgrew their PCL-scaffold-only counterparts and the control group in terms of neo-bone formation. By 6 weeks, PCL-silk scaffolds had 47.4–50.3% of bone growth that was twice as high as PCL scaffolds alone (16.7–19.9%). Similarly, by 12 weeks, the PCL-silk group had four times more (80–87.3%) new bone tissue production than the PCL group (18.6–22.4%). The promise of silk fibroin-reinforced PCL biomaterial for pre-clinical and clinical studies for craniofacial reconstructive applications is thus supported by these results.

Effect of Porcine- and Bovine-Derived Xenografts with Hydroxypropyl Methylcellulose for Bone Formation in Rabbit Calvaria Defects

In this study, hydroxypropyl methylcellulose (HPMC) was mixed with particle-type xenografts, derived from two different species (bovine and porcine), to increase the manipulability of bone grafts and compare the bone regeneration ability. Four circular defects with a diameter of 6 mm were formed on each rabbit calvaria, and the defects were randomly divided into three groups: no treatment (control group), HPMC-mixed bovine xenograft (Bo-Hy group), and HPMC-mixed porcine xenograft (Po-Hy group). At eight weeks, micro-computed tomography (µCT) scanning and histomorphometric analyses were performed to evaluate new bone formation within the defects. The results revealed that the defects treated with the Bo-Hy and the Po-Hy showed higher bone regeneration than the control group (p < 0.05), while there was no significant difference between the two xenograft groups (p > 0.05). Within the limitations of the present study, there was no difference in new bone formation between porcine and bovine xenografts with HPMC, and bone graft material was easily moldable with the desired shape during surgery. Therefore, the moldable porcine-derived xenograft with HPMC used in this study could be a promising substitute for the currently used bone grafts as it exhibits good bone regeneration ability for bony defects.

Fabrication of Low-Molecular-Weight Hyaluronic Acid-Carboxymethyl Cellulose Hybrid to Promote Bone Growth in Guided Bone Regeneration Surgery: An Animal Study.

Guided bone regeneration surgery is an important dental operation used to regenerate enough bone to successfully heal dental implants. When this technique is performed on maxilla sinuses, hyaluronic acid (HLA) can be used as an auxiliary material to improve the graft material handling properties. Recent studies have indicated that low-molecular hyaluronic acid (L-HLA) provides a better regeneration ability than high-molecular-weight (H-HLA) analogues. The aim of this study was to fabricate an L-HLA-carboxymethyl cellulose (CMC) hybrid to promote bone regeneration while maintaining viscosity. The proliferation effect of fabricated L-HLA was tested using dental pulp stem cells (DPSCs). The mitogen-activated protein kinase (MAPK) pathway was examined using cells cultured with L-HLA combined with extracellular-signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 inhibitors. The bone growth promotion of fabricated L-HLA/CMC hybrids was tested using an animal model. Micro-computer tomography (Micro-CT) and histological images were evaluated quantitatively to compare the differences in the osteogenesis between the H-HLA and L-HLA. Our results show that the fabricated L-HLA can bind to CD44 on the DPSC cell membranes and affect MAPK pathways, resulting in a prompt proliferation rate increase. Micro CT images show that new bone formation in rabbit calvaria defects treated with L-HLA/CMC was almost two times higher than in defects filled with H-HLA/CMC (p < 0.05) at 4 weeks, a trend that remained at 8 weeks and was confirmed by HE-stained images. According to these findings, it is reasonable to conclude that L-HLA provides better bone healing than H-HLA, and that the L-HLA/CMC fabricated in this study is a potential candidate for improving bone healing efficiency when a guided bone regeneration surgery was performed.

Microscopic analysis of the repair of critical bone defects in rabbits calvaria after the use of particulated autogenous bone or particulate autogenous bone associated with inorganic biomaterial

The aim of this study was to evaluate, through microscopic analysis, the repair of critical defects in rabbit calvaria after the use of particulate autogenous bone or particulate autogenous bone associated with inorganic biomaterial. Six male albino rabbits, New Zealand, were used, in which 4 defects were performed in each calvaria and randomly divided into 4 equitable groups: Group I – performed bone defect; Group II – performed bone defect and filled with particulate autogenous bone; Group III – performed bone defect and filled with particulate inorganic material (Bio-Oss®, Geistlich from Brazil); Group IV – performed bone defect and filled with particulate autogenous bone associated with particulate inorganic material (Bio-Oss® - Geistlich from Brazil) with proportion being 20:80. The analyze time of the procedure was 60 days. The microscopic analyze allowed to conclude that the autogenous, the Bio-Oss® and the Bio-Oss® associated with autogenous bone groups showed a bone neoformation on the defects, characterizing the material osteoconductive. However, the complete defect closure occurred on the autogenous bone and the autogenous bone associated with the inorganic bone, showing that the presence of the autogenous bone improves the characteristics of the biomaterial. The use of inorganic bone associated with the autogenous bone allowed a complete bone neoformation of the critic defect in the calvaria of rabbits.

Bone regeneration of demineralized dentin matrix with platelet-rich fibrin and recombinant human bone morphogenetic protein-2 on the bone defects in rabbit calvaria

BackgroundThis study was to evaluate the bone formation ability of demineralized dentin matrix (DDM) combined with platelet-rich fibrinogen (PRF) and DDM combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) to improve the osteoinductive ability of DDM.MethodsAfter four bone defects with a diameter of 8mm were created in the calvarium of each rabbit, DDM was grafted into the first defect (experimental groups 1), a combination of DDM and PRF was grafted into the second defect (experimental groups 2), and DDM with absorbed rhBMP-2 was grafted into the third defect (experimental groups 3). The fourth defect was used as the control group. Twelve healthy male rabbits (New Zealand, white rabbits) weighing around 3.0–4.0 kg were used. Among 12 rabbits, 3 rabbits were sacrificed immediately after surgery and at 2, 4, and 8 weeks after surgery, respectively. Histopathologic analysis and histomorphometric analysis were conducted to evaluate bone formation in each group.ResultsThe PRF/DDM group did not show a significantly higher degree of new bone formation in calvarial bone defects than the DDM group at 2, 4, and 8 weeks postoperatively in histopathological findings and histomorphometric results. On the other side, the rhBMP-2/DDM group showed higher degrees of new bone formation and calcification, and the lamellae of bone matrix, which are observed in mature bone tissue, were more distinctly visible in the rhBMP-2/DDM group. Moreover, the rhBMP-2/DDM group showed a significantly higher amount of new bone formation, compared to the DDM group at 4 and 8 weeks postoperatively (P<0.05) in histomorphometric results.ConclusionThe DDM has great potential as a carrier for the maintenance and sustained release of rhBMP-2, which has been recently receiving wide attention as a type of signaling molecules to promote bone formation.

A novel hydroxyapatite fiber material for the regeneration of critical-sized rabbit calvaria defects

Hydroxyapatite (HA) [Ca10 (PO4)6 (OH)2] has a high degree of chemical similarity with the mineral composition of animal bone. Hydroxyapatite fiber scaffold (HAF) is a biological material with a highly interconnected porous structure. We aimed to study the physical and biological characteristics of HAF and compare the osteogenic effects of HAF, natural osteogenic materials (NOM), and carbonate apatite (CO3Ap-DP) in the parietal defects of a rabbit's skull. X-ray analysis and histological assessment showed that HAF followed a trend of early initial osteogenesis and bone trabecular structure formation, especially at the cortical bone portion.Compared to the other two materials, HAF was more absorptive. Results indicated that HAF had the same osteoconductive and new bone formation properties as NOM and CO3Ap-DP. These findings will provide options for future material development and novel protocols for use in surgeries, ultimately leading to better patient outcomes.

Comparative Evaluation of Bone Repair with Four Different Bone Substitutes in Critical Size Defects

This study evaluated the osteoconductive potential of four biomaterials used to fill bone defects. For this, 24 male Albino rabbits were submitted to the creation of a bilateral 8 mm calvarial bone defect. The animals were divided into four groups—bovine hydroxyapatite, Bio-Oss® (BIO); Lumina-Bone Porous® (LBP); Bonefill® (BFL); and an alloplastic material, Clonos® (CLN)—and were euthanized at 14 and 40 days. The samples were subjected to histological and histometric analysis for newly formed bone area. Immunohistochemical analysis for Runt-related transcription factor 2 (Runx2), vascular endothelial growth factor (VEGF), and osteocalcin (OC) was performed. After statistical analysis, the CLN group showed greater new bone formation (NB) in both periods analyzed (p < 0.05). At 14 days, the NB showed greater values in BIO in relation to LBP and BFL groups; however, after 40 days, the LBP group surpassed the results of BIO (p < 0.001). The immunostaining showed a decrease in Runx2 intensity in BIO after 40 days, while it increased for LBP (p < 0.05). The CLN showed increased OC compared to the other groups in both periods analyzed (p < 0.05). Therefore, CLN showed the best osteoconductive behavior in critical defects in rabbit calvaria, and BFL showed the lowest osteoconductive property.

Comparison of Different Bone Filling Materials and Resorbable Membranes by Means of Micro-Tomography. A Preliminary Study in Rabbits.

The purpose of this work was to evaluate the behavior of different membranes and bone filling materials used to fill critical defects in rabbit calvaria. Four defects were prepared in the cranial calvaria of female rabbits. They were randomly divided into three subgroups according to the type of barrier membrane to be used. Four animals carried cross-linked bovine collagen membranes (Mem-Lok, Bio-Horizons, Birmingham, AL, USA)), four human fascia lata membranes (Tissue, Inbiomed SA, Córdoba, Argentina) and four human chorioamniotic membranes (Tissue. Inbiomed SA, Córdoba, Argentina). The defects were filled with the deproteinized bovine bone particulate Bio­Oss® (Geistlich­Pharma AG, Wolhusen, Switzerland), with particulate human hydroxyapatite MinerOss® (Bio-Horizons, Birmingham, AL, USA), with particulate dental material (Tissue Bank Foundation, Inbiomed S.A., Córdoba, Argentina), and the last one was left without the addition of filler material. In the first group of four specimens, a resorbable cross-linked bovine collagen membrane was placed over the skull and defects, without additional fixing. In the second group, a human fascia lata membrane was placed, without additional fixing. In the third group, a human chorioamniotic membrane was placed, without additional fixing. The animals were sacrificed at 4 and 8 weeks. The highest percentages of relative radiological density (average) were recorded considering the amnio-chorionic membranes (83.63%) followed by collagen (81.44%) and finally the fascia lata membranes (80.63%), but the differences were not statistically significant (p > 0.05). The sites grafted with a decellularized tooth (96.83%) and Bio­Oss (88.42%), recorded the highest percentages of radiological density but did not differ significantly from each other (subset 2). The three membranes used did not show statistical differences between them, in any of the two time periods used. There were statistical differences between the filling materials evidencing the presence of a large quantity of calcified material in the defects treated with particulate tooth and deproteinized bovine bone and while smaller amounts of calcified material were registered in the case of defects treated with human hydroxyapatite and those that were not treated.

Bone Healing in Rabbit Calvaria Defects Using a Synthetic Bone Substitute: A Histological and Micro-CT Comparative Study.

Bioactive alloplastic materials, like beta-tricalcium phosphate (β-TCP) and calcium sulfate (CS), have been extensively researched and are currently used in orthopedic and dental bone regenerative procedures. The purpose of this study was to compare the performance of EthOss versus a bovine xenograft and spontaneous healing. The grafting materials were implanted in standardized 8 mm circular bicortical bone defects in rabbit calvariae. A third similar defect in each animal was left empty for natural healing. Six male rabbits were used. After eight weeks of healing, the animals were euthanized and the bone tissue was analyzed using histology and micro-computed tomography (micro-CT). Defects treated with β-TCP/CS showed the greatest bone regeneration and graft resorption, although differences between groups were not statistically significant. At sites that healed spontaneously, the trabecular number was lower (p < 0.05) and trabecular separation was higher (p < 0.05), compared to sites treated with β-TCP/CS or xenograft. Trabecular thickness was higher at sites treated with the bovine xenograft (p < 0.05) compared to sites filled with β-TCP/CS or sites that healed spontaneously. In conclusion, the novel β-TCP/CS grafting material performed well as a bioactive and biomimetic alloplastic bone substitute when used in cranial defects in this animal model.

Novel cellulose/hydroxyapatite scaffolds for bone tissue regeneration: In vitro and in vivo study.

Cellulose scaffolds containing nano- or micro-hydroxyapatite (nHA or μHA) were prepared by the regeneration of cellulose from its acetylated derivative and the mechanical immobilization of inorganic particles, followed by freeze-drying. Microtomographic (micro-computed tomography) evaluation revealed that both scaffolds presented a highly interconnected porous structure, with a mean pore diameter of 490±94 and 540±132μm for cellulose/nHA and cellulose/μHA, respectively. In vitro and in vivo characterizations of the developed scaffolds were investigated. Commercially available bone allograft was used as a control material. For the in vitro characterization, osteoblastic cell cultures were used and characterized over time to evaluate cell adhesion, metabolic activity, and functional output (alkaline phosphatase activity and osteoblastic gene expression). The results revealed greater spreading cell distribution alongside an increased number of filopodia, higher MTT values, and significantly increased expression of osteoblastic genes (Runx-2, alkaline phosphatase, and BMP-2) for cellulose/nHA, compared with cellulose/μHA and the control. The in vivo biocompatibility was evaluated in a rabbit calvarial defect model. The investigated scaffolds were implanted in circular rabbit calvaria defects. Four- and 12-week bone biopsies were investigated using micro-computed tomography and histological analysis. Although both cellulose/HA scaffolds outperformed the assayed control, a significantly higher amount of newly formed mineralized tissue was found within the defects loaded with cellulose/nHA. Within the limitations of this study, the developed cellulose/HA scaffolds showed promising results for bone regeneration applications. The biological response to the scaffold seems to be greatly dependent on the HA particles' characteristics, with cellulose scaffolds loaded with nHA eliciting an enhanced bone response.

Controlled release of clarithromycin from PLGA microspheres enhances bone regeneration in rabbit calvaria defects.

This study evaluated the sustained release effect of clarithromycin-loaded in PLGA microspheres in a rabbit calvaria defect model. Four bone defects (ø5.0) were created in the calvaria of New Zealand White rabbits (n = 21, n = 7/time point). The defects were randomly designated to four groups. Group 1: No augmentation (sham), Group 2: beta-tricalcium phosphate (β-TCP), Group 3: β-TCP with 0.12 µg clarithromycin, and Group 4: β-TCP with 6.12 µg PLGA microspheres loaded with 0.12 µg Clarithromycin. After 2, 4, and 12 weeks of healing, bone regeneration was evaluated using micro-computed tomography (µCT) and histology. Clarithromycin release from PLGA microspheres revealed sustained release for around 4 weeks with ∼50% release during the first week. Histologically, new bone formation was evident at 2 and 4 weeks of healing in all groups and bone formation increased as a function of healing time. At 12 weeks, Group 4 showed significantly higher amount of newly formed bone compared to Group 1. The µCT showed that Group 4 expressed significantly higher bone formation compared to Group 1 at all time points. The in vivo findings showed that β-TCP with clarithromycin-loaded microspheres can enhance bone formation in bone defects. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 201-208, 2018.

Biphasic β-TCP mixed with silicon increases bone formation in critical site defects in rabbit calvaria.

The aim of this study was to assess the bone regeneration of critical size defects in rabbit calvarias filled with β-TCP doped with silicon. Twenty-one New Zealand rabbits were used in this study. Two critical size defects were created in the parietal bones. Three experimental groups were evaluated: Test A (HA/β-TCP granules alone), Test B (HA/β-TCP granules plus 3% silicon), Control (empty defect). The animals were sacrificed at 8 and 12 weeks. Evaluation was performed by μCT analysis and histomorphometry. μCT evaluation showed higher volume reduction in Test A group compared with Test B (P < 0.05). The Test B group showed the highest values for cortical closure and bone formation around the particles, followed by Test A and controls (P < 0.05). Within the limitations of this animal study, it can be concluded that HA/β-TCP plus 3% silicon increases bone formation in critical size defects in rabbit calvarias, and the incorporation of 3% silicon reduces the resorption rate of the HA/β-TCP granules.

Influence of different particles sizes of anorganic bovine bone matrix on bone repair process: histologic and radiographic analysis of surgically created defects in rabbit calvaria

Influence of different particles sizes of anorganic bovine bone matrix on bone repair process: histologic and radiographic analysis of surgically created defects in rabbit calvaria

Surface modification of PCL‐TCP scaffolds in rabbit calvaria defects: Evaluation of scaffold degradation profile, biomechanical properties and bone healing patterns

Traditionally, polycaprolactone (PCL) based scaffolds tend to degrade at a slow rate. Pretreatment of polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds under alkaline conditions can be utilized to increase the degradation rate and improve mechanical properties. Three groups of PCL-TCP scaffolds with varying pretreatment exposures with sodium hydroxide (NaOH) were studied in a rabbit calvaria defect model and analyzed at 2, 4, 8, 12, and 24 weeks. (Group A: Untreated, Group B: 3 M NaOH/ 48 h and Group C: 3 M NaOH/96 h). Micro-CT analysis demonstrated that scaffolds with increased surface roughness (Groups B and C) showed a greater impact on the overall volume loss during the early healing period between 2 and 8 weeks as compared to the untreated group. In addition, greater bone formation was detected in NaOH treated scaffolds as compared to the untreated group throughout the experiment. Scaffolds with increased surface roughness generally reported higher push out test and compressive strength values from 4 to 8 weeks of early healing. Interestingly, the mechanical properties displayed a decline in values from 12 weeks onwards in the modified groups suggesting a favorable breakdown or weakening of PCL-TCP scaffolds tailored for replacement by new bone formation.

Bone Formation on the Apatite-coated Zirconia Porous Scaffolds within a Rabbit Calvarial Defect

Previously, a strong and bioactive ceramic scaffold consisting of a porous zirconia body coated with apatite double layers (fluorapatite (FA) as an inner layer and hydroxyapatite (HA) as an outer layer) was successfully fabricated. In this contribution, the authors investigate the in vivo performance of the engineered bioceramic scaffolds using a rabbit calvarial defect model. In particular, the porosity and pore size of the scaffolds are varied in order to observe the geometrical effects of the scaffolds on their bone formation behaviors. The scaffolds supported on a zirconia framework can be produced with an extremely high porosity (approximately 84-87%), while retaining excellent compressive strength (approximately 7-8 MPa), which has been unachievable in the case of pure apatite scaffolds (approximately 74% porosity with approximately 2 MPa strength). The experimental groups used in this study include three types of zirconia scaffolds coated with apatite; high porosity (approximately 87%) with large pore size (approximately 500- 700 microm): AZ-HL, high porosity (approximately 84%) with small pore size (approximately 150-200 microm): AZ-HS, and low porosity (approximately 75%) with large pore size (approximately 500-700 microm): AZ-LL, as well as one type of HA porous scaffold: low porosity (approximately 74%) with a large pore size (approximately 500-700 microm) for the purpose of comparison. The scaffolds prepared with dimensions of approximately 10 mm (diameter) x 1.2 mm (thickness) are grafted in rabbit calvaria defects. The histological sections are made at 4 and 12 weeks after surgery and immunohistochemical analyses are performed on the samples. All of the specimens show a good healing response without adverse tissue reactions. Good healing is shown at 4 weeks post-surgery with the ingrowth of new bone into the macropore-channels of the scaffolds. The newly formed bone amounts to approximately 19.9-24.2% of the initial defect area, depending on the scaffold type, but there is no statistical significance between the scaffold groups. However, the defects without the scaffolds (control group) show a significantly lower bone formation ratio (approximately 4.3%). At twelve weeks after surgery, the extent of new bone formation is more pronounced in all of the scaffold groups. All of the scaffold groups show significantly higher bone formation ratios (26.7-46.9%) with respect to the control without the graft. In the comparison between the scaffold groups, those with high porosities (AZ-HL and AZ-HS) exhibit significantly higher bone formation as compared to the scaffold with low porosity (AZ-LL). Based on the present in vivo test performed within a rabbit calvaria defect model, it is concluded that the apatite-coated zirconia scaffolds show good bone forming ability and are considered to be a promising scaffolding material for bone regeneration since they possess a high level of both mechanical and biological properties.