Bilateral Calvarial Defects Research Articles

Fabrication and In Vivo Assessment of Oxidatively Responsive PolyHIPE Scaffolds for Use in Diabetic Orthopedic Applications.

Achieving surgical success in orthopedic patients with metabolic disease remains a substantial challenge. Diabetic patients exhibit a unique tissue microenvironment consisting of high levels of reactive oxygen species (ROS), which promotes osteoclastic activity and leads to decreased bone healing. Alternative solutions, such as synthetic grafts, incorporating progenitor cells or growth factors, can be costly and have processing constraints. Previously, the potential for thiol-methacrylate networks to sequester ROS while possessing tunable mechanical properties and degradation rates has been demonstrated. In this study, the ability to fabricate thiol-methacrylate interconnected porous scaffolds using emulsion templating to create monoliths with an average porosity of 97.0% is reported. The average pore sizes of the scaffolds range from 27 to 656µm. The scaffolds can sequester pathologic levels of ROS via hydrogen peroxide consumption and are not impacted by sterilization. Subcutaneous implantation shows no signs of acute toxicity. Finally, in a 6-week bilateral calvarial defect model in Zucker diabetic fatty rats, ROS scaffolds increase new bone volume by 66% over sham defects. Histologic analysis identifies woven bone infiltration throughout the scaffold and neovascularization. Overall, this study suggests that porous thiol-methacrylate scaffolds may improve healing for bone grafting applications where high levels of ROS hinder bone growth.

Evaluation of the time-dependent osteogenic activity of glycerol incorporated magnesium oxide nanoparticles in induced calvarial defects

IntroductionMagnesium-based biomaterials have been explored for their potential as bone healing materials, as a result of their outstanding biodegradability and biocompatibility. These characteristics make magnesium oxide nanoparticles (MgO NPs) a promising material for treating bone disorders. The purpose of this investigation is to assess the osteogenic activity of newly-developed locally administered glycerol-incorporated MgO NPs (GIMgO NPs) in rabbits’ calvarial defects. Materials and methodsCharacterization of GIMgO was done by X-ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FTIR). Bilateral calvarial defects were created in eighteen New Zealand Rabbits, of which they were divided into 3 groups with time points corresponding to 2, 4, and 6 weeks postoperatively (n = 6). One defect was implanted with absorbable gel foam impregnated with GIMgO NPs while the other was implanted with gel foam soaked with glycerol (the control). The defects were assessed using histological, Micro-Computed Tomography (Micro-CT), and histometric evaluation. ResultsThe characterization of the GIMgO nanogel revealed the presence of MgO NPs and glycerol as well as the formation of the crystalline phase of the MgO NPs within the nanogel sample. The histological and micro-CT analysis showed time-dependent improvement of healing activity in the calvarial defects implanted with GIMgO NPs when compared to the control. Furthermore, the histometric analysis demonstrated a marked increase in the total area of new bone, connective tissue, new bone area and volume in the GIMgO NPs implanted site. Statistically, the amount of new bone formation was more significant at 6 weeks than at 2 and 4 weeks postoperatively in the calvarial defects implanted with GIMgO NPs as compared to the control. ConclusionThe locally applied GIMgO NPs demonstrated efficacy in promoting bone formation, with more significant effects observed over an extended period. These findings suggest its suitability for clinical use as a therapeutic alternative to enhance bone healing.

The efficacy of a semi-resorbable membrane based on silk fibroin-Glycerol on bone regeneration in rabbit calvarial defects compared to a commercial collagen membrane.

Silk fibroin-glycerol-based membranes were fabricated and characterized for use as a self-maintaining and non-collapsible semi-resorbable membrane in guided bone regeneration. The study assessed the bone regeneration capacity of silk fibroin-glycerol-based membranes compared to a collagen membrane in 10-mm circular bilateral calvarial defects of 20 male New Zealand white rabbits. The animals were divided into two sets of time frames of 4 and 12 weeks and allocated into four groups (n = 5/group); an empty defect (E), a collagen membrane (Bio-Gide®; BG), a silk fibroin-glycerol-collagen membrane (SGC), and a silk fibroin-glycerol membrane (SG). The bone density (optical density, OD) from the 2D radiographs, tissue reaction from histological sections, new bone volume, and area from micro-CT and the histomorphometry were evaluated. The Mean OD of the E (34.49 ± 14.21%) and BG groups (35.71 ± 9.65%) at 12 weeks were higher than at 4 weeks, but the SGC (39.04 ± 7.94%) and SG (40.96 ± 9.25%) groups were lower at 4 weeks. The new bone volumes at 4 weeks of the SG (24.19 ± 1.35%) and SGC groups (24.19 ± 3.47%) were significantly higher than the BG group (16.93 ± 2.95%) but were not different from the E group (18.39±4.78%). At 12 weeks, the new bone volumes in the SGC (29.09 ± 3.81%), SG (29.11 ± 5.94%), and BG groups (26.26 ± 4.42%) were higher than in the E group (21.63 ± 5.81%) without statistical significance. Histological images in the SGC and SG groups showed slow biodegradation without a foreign body reaction. The new bone area at 4 weeks was lowest in BG (12.95 ± 5.44%), and the others were comparable. At 12 weeks, the new bone area in the E group (23.55±8.69%) was lower than the BG (31.42 ± 6.18%), SG (35.25 ± 13.92%), and SGC groups (36.35 ± 10.23%). Silk fibroin-glycerol-based membranes are semi-resorbable membranes that possess a self-maintaining property, have a barrier function without collapsing, and are successful in facilitating bone regeneration.

A Bilayer Membrane Doped with Struvite Nanowires for Guided Bone Regeneration.

Guided bone regeneration (GBR) therapy demonstrates a prominent curative effect on the management of craniomaxillofacial (CMF) bone defects. In this study, a GBR membrane consisting of a microporous layer and a struvite-nanowire-doped fibrous layer is constructed via non-solvent induced phase separation, followed by an electrospinning procedure to treat critical-sized calvarial defects. The microporous layer shows selective permeability for excluding the rapid-growing non-osteogenic tissues and potential wound stabilization. The nanowire-like struvite is synthesized as the deliverable therapeutic agent within the fibrous layer to facilitate bone regeneration. Such a membrane displays a well-developed heterogeneous architecture, satisfactory mechanical performance, and long-lasting characteristics. The in vitro biological evaluation reveals that apart from being a strong barrier, the bilayer struvite-laden membrane can actively promote cellular adhesion, proliferation, and osteogenic differentiation. Consequently, the multifunctional struvite-doped membranes are applied to treat 5mm-sized bilateral calvarial defects in rats, resulting in overall improved healing outcomes compared with the untreated or the struvite-free membrane-treated group, which is characterized by enhanced osteogenesis and significantly increased new bone formation. The encouraging preclinical results reveal the great potential of the bilayer struvite-doped membrane as a clinical GBR device for augmenting large-area CMF bone reconstruction.

Periodontal Ligament Stem Cell-Derived Small Extracellular Vesicles Embedded in Matrigel Enhance Bone Repair Through the Adenosine Receptor Signaling Pathway.

PurposeSmall extracellular vesicles (sEVs) are natural biocarriers for biomolecule transfer between cells and promising therapeutic strategies for bone defect repair. In this study, human periodontal ligament stem cell (PDLSC)-derived sEVs (P-EVs) were immobilized in Matrigel to establish a topical cell-free transplantation strategy for bone repair.MethodsPDLSCs were cultured and P-EVs were isolated from the culture supernatant. In a rat bilateral calvarial defect model, P-EV/Matrigel was plugged into one defect and PBS/Matrigel was applied to the other. Bone repair in vivo was assessed by micro-computed tomography, histomorphometry, and immunohistochemical staining. In vitro, we investigated the effects of P-EVs on the proliferation and migration capabilities of bone marrow mesenchymal stem cells (BMMSCs) and explored the potential mechanism of action.ResultsThe in vivo study showed that P-EV/Matrigel accelerated bone tissue repair by increasing cell infiltration when compared with the control. In vitro, P-EVs enhanced proliferation and migration of BMMSCs via increased phosphorylation of AKT and extracellular signal-regulated kinase 1/2 (ERK1/2). The role of P-EV-induced adenosine receptor signaling in AKT and ERK1/2 phosphorylation was a key mediator during enhanced BMMSC migration.ConclusionThese results are the first to demonstrate that P-EVs accelerated the repair of bone defects, partially through promoting cell proliferation and migration. P-EV/Matrigel, which combines topical EV-implantation and extracellular matrix scaffolds, provides a new cell-free strategy for bone tissue repair.

Evaluation of a self-fitting, shape memory polymer scaffold in a rabbit calvarial defect model

Self-fitting scaffolds prepared from biodegradable poly(ε-caprolactone)-diacrylate (PCL-DA) have been developed for the treatment of craniomaxillofacial (CMF) bone defects. As a thermoresponsive shape memory polymer (SMP), with the mere exposure to warm saline, these porous scaffolds achieve a conformal fit in defects. This behavior was expected to be advantageous to osseointegration and thus bone healing. Herein, for an initial assessment of their regenerative potential, a pilot in vivo study was performed using a rabbit calvarial defect model. Exogenous growth factors and cells were excluded from the scaffolds. Key scaffold material properties were confirmed to be maintained following gamma sterilization. To assess scaffold integration and neotissue infiltration along the defect perimeter, non-critically sized (d = 8 mm) bilateral calvarial defects were created in 12 New Zealand white rabbits. Bone formation was assessed at 4 and 16 weeks using histological analysis and micro-CT, comparing defects treated with an SMP scaffold (d = 9 mm x t = 1 or 2 mm) to untreated defects (i.e. defects able to heal without intervention). To further assess osseointegration, push-out tests were performed at 16 weeks and compared to defects treated with poly(ether ether ketone) (PEEK) discs (d = 8.5 mm x t = 2 mm). The results of this study confirmed that the SMP scaffolds were biocompatible and highly conducive to bone formation and ingrowth at the perimeter. Ultimately, this resulted in similar bone volume and surface area versus untreated defects and superior performance in push-out testing versus defects treated with PEEK discs. Statement of significanceCurrent treatments of craniomaxillofacial (CMF) bone defects include biologic and synthetic grafts but they are limited in their ability to form good contact with adjacent tissue. A regenerative engineering approach using a biologic-free scaffold able to achieve conformal fitting represents a potential “off-the-shelf” surgical product to heal CMF bone defects. Having not yet been evaluated in vivo, this study provided the preliminary assessment of the bone healing potential of self-fitting PCL scaffolds using a rabbit calvarial defect model. The study was designed to assess scaffold biocompatibility as well as bone formation and ingrowth using histology, micro-CT, and biomechanical push-out tests. The favorable results provide a basis to pursue establishing self-fitting scaffolds as a treatment option for CMF defects.

Bone Regeneration Using PEVAV/β-Tricalcium Phosphate Composite Scaffolds in Standardized Calvarial Defects: Micro-Computed Tomographic Experiment in Rats.

Bone regeneration using beta-tricalcium phosphate (β-TCP) can be practiced using a biocomposite scaffold. Poly(ethylene-co-vinylalcohol)/poly(δ-valerolactone)/β-tricalcium phosphate (PEVAV/β-TCP) composite scaffolds showed promising in vitro results. This study evaluated the bone regenerative potential of PEVAV/β-TCP biocomposite scaffolds in standardized calvarial defects in a rat model over 4 and 10 weeks. Bilateral calvarial defects (5 mm in diameter and about 1.5 mm thick, equivalent to the thickness of the calvaria) were created in 40 male Wistar albino rats. The defects were grafted with either commercially available β-TCP (positive control), PEVAV/β-TCP 70, or PEVAV/β-TCP 50, or left empty (negative control), depending on the group to which the animal was randomly assigned, to be covered before flap closure with resorbable collagen membrane (RCM). At 4 and 10 weeks post-surgery, the collected rat calvaria were evaluated using micro computed tomography (micro-CT) analysis, to assess the newly formed bone volume (NFBV), newly formed bone mineral density (NFBMD), and remaining graft volume (RGV). The results showed that calvarial defects grafted with the PEVAV/β-TCP biocomposite exhibited higher NFBV than did control defects, both at 4 and 10 weeks post-surgery. Furthermore, calvarial defects grafted with PEVAV/β-TCP 70 showed the highest NFBV among all grafting conditions, with a statistically significant difference recorded at 10 weeks post-surgery. The PEVAV/β-TCP composite scaffold showed potentiality for the regeneration of critical-sized calvarial bone defects in a rat model.

Three-Dimensionally-Printed Bioactive Ceramic Scaffolds: Construct Effects on Bone Regeneration.

The utilization of three-dimensionally (3D)-printed bioceramic scaffolds composed of beta-tricalcium phosphate in conjunction with dipyridamole have shown to be effective in the osteogenesis of critical bone defects in both skeletally immature and mature animals. Furthermore, previous studies have proven the dura and pericranium's osteogenic capacity in the presence of 3D-printed scaffolds; however, the effect galea aponeurotica on osteogenesis in the presence of 3D scaffolds remains unclear. Critical-sized (11 mm) bilateral calvarial defects were created in 35-day old rabbits (n = 7). Two different 3D scaffolds were created, with one side of the calvaria being treated with a solid nonporous cap and the other with a fully porous cap. The solid cap feature was designed with the intention of preventing communication of the galea and the ossification site, while the porous cap permitted such communication. The rabbits were euthanized 8 weeks postoperatively. Calvaria were analyzed using microcomputed tomography, 3D reconstruction, and nondecalcified histologic sectioning in order assess differences in bone growth between the two types of scaffolding. Scaffolds with the solid (nonporous) cap yielded greater percent bone volume (P = 0.012) as well as a greater percent potential bone (P = 0.001) compared with the scaffolds with a porous cap. The scaffolds with porous caps also exhibited a greater percent volume of soft tissue (P < 0.001) presence. There were no statistically significant differences detected in scaffold volume. A physical barrier preventing the interaction of the galea aponeurotica with the scaffold leads to significantly increased calvarial bone regeneration in comparison with the scaffolds allowing for this interaction. The galea's interaction also leads to more soft tissue growth hindering the in growth of bone in the porous-cap scaffolds.

Bone Tissue Engineering in the Growing Calvaria Using Dipyridamole-Coated, Three-Dimensionally-Printed Bioceramic Scaffolds: Construct Optimization and Effects on Cranial Suture Patency.

Three-dimensionally-printed bioceramic scaffolds composed of β-tricalcium phosphate delivering the osteogenic agent dipyridamole can heal critically sized calvarial defects in skeletally mature translational models. However, this construct has yet to be applied to growing craniofacial models. In this study, the authors implanted three-dimensionally-printed bioceramic/dipyridamole scaffolds in a growing calvaria animal model and evaluated bone growth as a function of geometric scaffold design and dipyridamole concentration. Potential adverse effects on the growing suture were also evaluated. Bilateral calvarial defects (10 mm) were created in 5-week-old (approximately 1.1 kg) New Zealand White rabbits (n = 16 analyzed). Three-dimensionally-printed bioceramic scaffolds were constructed in quadrant form composed of varying pore dimensions (220, 330, and 500 μm). Each scaffold was coated with collagen and soaked in varying concentrations of dipyridamole (100, 1000, and 10,000 μM). Controls consisted of empty defects. Animals were killed 8 weeks postoperatively. Calvariae were analyzed using micro-computed tomography, three-dimensional reconstruction, and nondecalcified histologic sectioning. Scaffold-induced bone growth was statistically greater than bone growth in empty defects (p = 0.02). Large scaffold pores, 500 μm, coated in 1000 μM dipyridamole yielded the most bone growth and lowest degree of scaffold presence within the defect. Histology showed vascularized woven and lamellar bone along with initial formation of vascular canals within the scaffold lattice. Micro-computed tomographic and histologic analysis revealed patent calvarial sutures without evidence of ectopic bone formation across all dipyridamole concentrations. The authors present an effective pediatric bone tissue-engineering scaffold design and dipyridamole concentration that is effective in augmentation of calvarial bone generation while preserving cranial suture patency.

Spatially controlled rhBMP-2 mediated calvarial bone formation in a transgenic mouse model

The study aimed to investigate the localized osteogenic activity of recombinant human bone morphogenetic protein (rhBMP-2), when delivered using enzymatically crosslinkable injectable glycol chitosan hydrogel. A critical sized bilateral calvarial defect model was used wherein one defect was implanted with rhBMP-2 loaded hydrogel (HPP-GC+BMP). The neighboring defect was implanted with an osteoconductive, collagen hydroxyapatite matrix “Healos®”. The implantation of HPP-GC+BMP led to complete closure of the critical sized defect at 4 weeks post-implantation. The neighboring site implanted with Healos® however, did not show any bone formation. The spatial control of rhBMP-2 bioactivity at the cellular level was confirmed by histological and histomorphometric analysis of the calvaria isolated from Col3.6 transgenic animals which can express fluorescence in osteoblast and pre-osteoblast cells. The retained rhBMP-2 in HPPGC+BMP implant was able to localize osteoprogenitor recruitment and osteogenesis, at the implantation site. The results demonstrate the efficacy of HPP-GC hydrogel in minimizing the diffusive loss of rhBMP-2 from the implantation site, compared to the collagen hydroxyapatite scaffold.

Osteogenic potential of recombinant human bone morphogenetic protein-9/absorbable collagen sponge (rhBMP-9/ACS) in rat critical size calvarial defects.

It has been reported that bone morphogenetic protein (BMP)-9 has potent osteoinductive properties among the BMP family by adenovirus-transfection experiments. We very recently reported that absorbable collagen sponge (ACS) as a carrier for recombinant human (rh) BMP-9, compared with chitosan sponge, was suitable for inducing bone healing/regeneration by BMP-9 in a rat calvarial defect model. The aim of this study was to evaluate different doses of rhBMP-9/ACS on new bone formation in rat critical size calvarial defects. Bilateral calvarial defects (n=32) were surgically created in 16 wistar rats and randomly filled with one of the following materials: (1) absorbable collagen sponge (ACS) alone; (2) 1μg-rhBMP-9/ACS (L-rhBMP-9/ACS); (3) 5μg-rhBMP-9/ACS (H-rhBMP-9/ACS); and (4) blank defects (control). The animals were sacrificed 8weeks postsurgery for radiographic and histomorphometric analyses. Bone volume and defect closure were statistically higher in the rhBMP-9/ACS-implanted (L-rhBMP-9/ACS and H-rhBMP-9/ACS) groups when compared with ACS-alone group (p<0.05). Furthermore, defects filled with H-rhBMP-9/ACS showed the highest levels of newly formed bone area (NBA) and NBA/total defect area among all groups. No significant differences in any of the radiographic and histometric parameters could be observed between both concentrations of rhBMP-9. Within the limits of this study, it can be concluded that rhBMP-9/ACS-induced bone formation can be reached with as little as 1μg/site in rat critical size calvarial defects. RhBMP-9 could be a potential therapeutic growth factor for future bone regenerative procedures.

Evaluation of the donor cell contribution in rhBMP-2 mediated bone formation with chitosan thermogels using fluorescent protein reporter mice.

Our current understanding regarding the contribution of donor cells in growth factor and cell based tissue regeneration strategies is limited. The present study attempts to utilize fluorescent protein reporter mice [Col3.6Topaz (enhanced yellow fluorescent protein, EYFP) as host and Col3.6Cyan (enhanced cyan fluorescent protein, ECFP) as donor] to determine donor cell contribution in bone regeneration using a bilateral calvarial defect model. Thermogelling chitosan hydrogels (Chi-AHP) were used as bone marrow stromal cell (BMSC) delivery vehicle in the presence and absence of recombinant human bone morphogenetic protein-2 (rhBMP-2). Co-delivery of rhBMP-2 and donor BMSCs led to a significant increase in bone formation indicating the potential of using the combination approach for improved regeneration. On a cellular level, presence of rhBMP-2 resulted in an increased host cell derived osteoblast infiltration at 8 weeks. However, the new mineralized tissue presented two distinct morphological features based on its cellular origin. Regenerated tissue associated with ECFP positive donor cells showed a woven bone-like structure with diffuse alizarin complexone (AC) label and minimal tartrate resistant acid phosphatase (TRAP) activity, indicating the presence of immature early osteoblast cells depositing mineralized tissue without progressing to a mature lamellar bone. Host derived bone showed sharp mineralization line (AC), strong alkaline phosphatase (ALP) and TRAP activity, indicating the presence of actively mineralizing and remodeling, mature lamellar bone matrix. The study demonstrated the remarkable potential of transgenic reporters to improve our understanding of donor cell contribution during bone formation.

Bone regeneration in calvarial defects in a rat model by implantation of human bone marrow-derived mesenchymal stromal cell spheroids.

Mesenchymal stem cell (MSC) condensation contributes to membrane ossification by enhancing their osteodifferentiation. We investigated bone regeneration in rats using the human bone marrow-derived MSC-spheroids prepared by rotation culture, without synthetic or exogenous biomaterials. Bilateral calvarial defects (8 mm) were created in nude male rats; the left-sided defects were implanted with MSC-spheroids, β-tricalcium phosphate (β-TCP) granules, or β-TCP granules + MSC-spheroids, while the right-sided defects served as internal controls. Micro-computed tomography and immunohistochemical staining for osteocalcin/osteopontin indicated formation of new, full-thickness bones at the implantation sites, but not at the control sites in the MSC-spheroid group. Raman spectroscopy revealed similarity in the spectral properties of the repaired bone and native calvarial bone. Mechanical performance of the bones in the MSC-implanted group was good (50 and 60 % those of native bones, respectively). All tests showed poor bone regeneration in the β-TCP and β-TCP + MSC-spheroid groups. Thus, significant bone regeneration was achieved with MSC-spheroid implantation into bone defects, justifying further investigation.

The effects of nonvascularized versus vascularized bone grafting on calvarial defect healing.

It remains unknown whether bone graft vascularity influences calvarial healing. The purposes of this study were (1) to develop a model to study nonvascularized and vascularized calvarial grafts as well as (2) to compare effects of bone graft vascularity on calvarial healing. Bilateral calvarial defects were created in 26 Wistar rats. The defects were left empty within 1 parietal region. On the contralateral side, the defects were partially closed with native parietal bone (control group, n = 6), nonvascularized (N-V, n = 10), or vascularized bone grafts (VAS, n = 10). The vascularized grafts were supplied by perforating dural arterioles. Bone mineralization and healing patterns from serial microcomputed tomographic scans were compared within and across the groups using parametric and nonparametric tests. Differences in bone mineral content across sides were significant between the groups at weeks 6 (P = 0.016) and 12 (P = 0.025). Bone formation was greater within both the control and VAS groups versus the N-V group at weeks 6 and 12 (P < 0.05). Healing patterns differed between the groups (P < 0.05), progressing through islands of new bone formation within the control and VAS groups while limited to defect margins on the N-V graft side. In conclusion, a bilateral calvarial defect model was established to study bone graft vascularity. Bone quantity and healing patterns differed in the presence of the nonvascularized versus vascularized grafts. Although the calvarial defect model is often applied within the plastic surgery literature to study bone substitutes, greater understanding of basic mechanisms influencing calvarial healing is first needed to avoid confounding results.

Fibroblast Growth Factor-2 Isoform (Low Molecular Weight/18 kDa) Overexpression in Preosteoblast Cells Promotes Bone Regeneration in Critical Size Calvarial Defects in Male Mice

Repair of bone defects remains a significant clinical problem. Bone morphogenetic protein 2 (BMP2) is US Food and Drug Administration-approved for fracture healing but is expensive and has associated morbidity. Studies have shown that targeted overexpression of the 18-kDa low-molecular-weight fibroblast growth factor 2 isoform (LMW) by the osteoblastic lineage of transgenic mice increased bone mass. This study tested the hypotheses that overexpression of LMW would directly enhance healing of a critical size calvarial bone defect in mice and that this overexpression would have a synergistic effect with low-dose administration of BMP2 on critical size calvarial bone defect healing. Bilateral calvarial defects were created in LMW transgenic male mice and control/vector transgenic (Vector) male mice and scaffold with or without BMP2 was placed into the defects. New bone formation was assessed by VIVA-computed tomography of live animals over a 27-week period. Radiographic and computed tomography analysis revealed that at all time points, healing of the defect was enhanced in LMW mice compared with that in Vector mice. Although the very low concentration of BMP2 did not heal the defect in Vector mice, it resulted in complete healing of the defect in LMW mice. Histomorphometric and gene analysis revealed that targeted overexpression of LMW in osteoblast precursors resulted in enhanced calvarial defect healing due to increased osteoblast activity and increased canonical Wnt signaling.

High-intensity Nd:YAG laser accelerates bone regeneration in calvarial defect models

High-power pulsed lasers have been recently regarded to be anabolic to bone, but in vivo evidence is still lacking. This study aimed to investigate the capacity of bone repair using a high-power, Q-switched, pulsed, neodymium-doped yttrium aluminium garnet (Nd:YAG) laser, using bilateral calvarial defect models having non-critical sized, 5 mm (rat) or 8 mm (rabbit) diameter. One of the bilateral defects, which were all filled with collagen sponge or left empty, was irradiated with a Nd:YAG laser once every 2 days for 2 weeks at a constant total fluence rate (344 J/cm(2) ), output power (0.75 W), pulse repetition rate (15 pps) and wavelength (1064 nm) and examined for the laser effect. The same experimental scheme was designed using a rabbit calvarial defect model implanted with sponge, which was explored for the dose effect of output power at 0.75 and 3 W with the same quantities of the other parameters. New bone formation was evaluated by micro-computed tomography-based analysis and histological observation at 4 weeks after surgery. Laser irradiation significantly increased new bone formation by approximately 45%, not only in the sponge-filled defects of rats but also when the defects were left empty, compared to the non-irradiated group. Consistently, both doses of output power (0.75 and 3 W) enhanced new bone formation, but there was no significant difference between the two doses. This study is one of the first to demonstrate the beneficial effect of Nd:YAG lasers on the regeneration of bone defects which were left empty or filled with collagen sponge, suggesting its great potential in postoperative treatment targeting local bone healing.

Bone Formation in Algae-Derived and Synthetic Calcium Phosphates With or Without Poloxamer

Calcium phosphate ceramics such as hydroxyapatite (HA) and biphasic calcium phosphates are used clinically to repair bone defects. These calcium phosphate ceramics can differ by composition, structure, and rate of degradation. This study compared 3 calcium phosphate ceramics, 2 of which have similar structure but different composition: 100% HA (algae derived) and HA/β-tricalcium phosphate (β-TCP) 20/80 (algae derived), and 2 with different structure but similar composition: HA/β-TCP 20/80 (algae derived) and HA/β-TCP 15/85 (synthetic). Calcium phosphate ceramics can be difficult to handle and contour during the surgeries. To improve handling, Poloxamer 407 (P407) was added to the 3 ceramics, and its effect on bone healing was also assessed. Bilateral calvarial defects created in the parietal bones of New Zealand white rabbits were left unfilled or were filled with autograft or one of the ceramics, with and without P407. Six weeks after operation, healing was evaluated qualitatively by histology and quantitatively by micro-computed tomography analysis and histomorphometry. All 3 calcium phosphate ceramics demonstrated osteoconductivity and performed similarly in supporting new bone formation, suggesting that the differences in their composition, structure, or degradation did not significantly affect their ability to promote bone healing in this application. Incorporating P407 did not impede osteoconductivity as HA and biphasic calcium phosphate combined with P407 performed similarly as when used alone for craniofacial defect repair.

Bone healing with an in situ–formed bioresorbable polyethylene glycol hydrogel membrane in rabbit calvarial defects

The aims of this study were to test whether or not the application of an in situ-formed synthetic polyethylene glycol hydrogel (PEG) used as a biodegradable membrane for guided bone regeneration with a variety of graft materials and ambient oxygen or hyperbaric oxygen (HBO) environments would result in enhanced bone regeneration, and to observe the histologic and histomorphometric aspects of bone healing of the calvarial defects with and without a PEG membrane. Thirty adult, skeletally mature, male New Zealand white rabbits were randomly divided into 3 groups of 10 animals each. Bilateral 15-mm-diameter critical-size defects were created in the parietal bones of each animal. Group 1 served as a control with unfilled bilateral calvarial defects, group 2 had bilateral calvarial defects filled with morcelized autogenous calvarial bone, and group 3 had bilateral calvarial defects filled with a biphasic calcium phosphate ceramic. One of the calvarial defects was randomly protected with a PEG resorbable liquid membrane in each animal. Five animals from each group underwent a course of HBO treatment (2.4 ATA 100% oxygen for 90 minutes 5 days a week for 4 weeks) and the other 5 served as control and did not receive any supplemental oxygen (normobaric). The animals were killed 6 weeks after their surgery, and their parietal bones were harvested. The specimens were analyzed with microscopic computerized tomography (microCT) scans and histomorphometrics. The unfilled normobaric control bony defects did not heal, proving the critical-size nature of these defects. The presence of autogenous bone or bone ceramic in the defects increased the bone volume fraction and bone mineral density of the defects (P < .001). The presence of a membrane in the ungrafted and autogenous bone grafted defects resulted in a decrease in the corrected bone volume fraction (P = .002) but not in the bone ceramic grafted defects (P = .580). Bony healing of defects where the membrane was unsupported was compromised; the membrane did not maintain the desired bone regeneration volume with the unfilled and autogenous bone grafted groups. The PEG resorbable liquid membrane worked best with the bone ceramic material. HBO did not ameliorate the healing of the autogenous bone graft or ceramic filled defects in the 6-week time period of this study. Although the PEG resorbable liquid membrane is easy to use and forms an occlusive layer, caution is recommended when using the membrane over an unsupported defect. HBO did not ameliorate bony healing with the membrane at the early 6-week time point. The authors recommend future assessment with HBO at the 12-week time point.

An Evaluation of Hydroxyapatite and Biphasic Calcium Phosphate in Combination With Pluronic F127 and BMP on Bone Repair

Calcium phosphates like hydroxyapatite (HA), beta-tricalcium phosphate (beta-TCP), and their mixtures (biphasic calcium phosphates; BCP) are used clinically to repair bone defects. These materials can be difficult to handle and have no inherent biological activity. Handling properties of other bone substitute materials have been improved by combining them with an inert carrier such as Pluronic F-127 (Pluronic, BASF, Mt. Olive, NJ), while the addition of bone morphogenetic proteins (BMP) with such implants has also been shown to enhance bone repair. This study assessed the impact of adding Pluronic and BMPs to an HA (C-Graft) or a BCP (80/20 HA/beta-TCP ratio; Algisorb) implant's ability of promote bony repair in the rabbit calvarial defect model.Twenty-five New Zealand white rabbits were divided into 5 groups of 5 animals each. Bilateral calvarial defects were made in the parietal bones of each animal. HA or BCP alone or combined with Pluronic and/or BMP were implanted into the defect sites. Animals were euthanized at 6 weeks, postoperatively. Bone regeneration was evaluated quantitatively by histomorphometry. The amount of bone regeneration, which occurred in defects containing HA and BCP, was similar over the time period studied. Incorporating Pluronic increased handling and moldability without compromising osteoconductivity of either calcium phosphate. The addition of BMP significantly increased the amount of new bone formed with all calcium phosphates studied (P < 0.05). These results suggest that Pluronic can be added to calcium phosphates to enhance handling and moldability without any negative effects on their biocompatibility and that healing can be enhanced with the incorporation of BMPs.

Poster 122: The Effects of Hyperbaric Oxygen on Bone Regeneration Using Bone Substitutes With or Without a Resorbable Liquid Membrane

Poster 122: The Effects of Hyperbaric Oxygen on Bone Regeneration Using Bone Substitutes With or Without a Resorbable Liquid Membrane