Bone healing and graft resorption of autograft, anorganic bovine bone and beta-tricalcium phosphate. A histologic and histomorphometric study in the mandibles of minipigs

OBJECTIVE: The purpose was to qualitatively and quantitatively compare the bone formation and graft resorption of two different bone substitutes used in both orthopedic and oral surgery, with autogenous bone as a positive control. MATERIALS AND METHODS: Three standardized bone defects were prepared in both mandibular angles of 12 adult minipigs. The defects were grafted with either autograft, anorganic bovine bone (ABB), or synthetic beta-tricalcium phosphate (beta-TCP). Sacrifice was performed after 1, 2, 4, and 8 weeks for histologic and histomorphometric analysis. RESULTS: At 2 weeks, more new bone formation was seen in defects filled with autograft than with ABB (P approximately 0.0005) and beta-TCP (P approximately 0.002). After 4 weeks, there was no significant difference between beta-TCP and the two other materials. Defects grafted with ABB still exhibited less bone formation as compared with autograft (P approximately 0.004). At 8 weeks, more bone formation was observed in defects grafted with autograft (P approximately 0.003) and beta-TCP (P approximately 0.00004) than with ABB. No difference could be demonstrated between beta-TCP and autograft. beta-TCP resorbed almost completely over 8 weeks, whereas ABB remained stable. CONCLUSION: Both bone substitutes seemed to decelerate bone regeneration in the early healing phase as compared with autograft. All defects ultimately regenerated with newly formed bone and a developing bone marrow. The grafting materials showed complete osseous integration. Both bone substitutes may have a place in reconstructive surgery where different clinical indications require differences in biodegradability.     

Calcium phosphate/poly(D,L-lactic-co-glycolic acid) composite bone substitute materials: evaluation of temporal degradation and bone ingrowth in a rat critical-sized cranial defect

Objectives: The present study aimed to provide temporal information on material degradation and bone formation using composite (C) bone defect filler materials consisting of calcium phosphate cement (CaP) and poly(d ,l ‐lactic‐co‐glycolic acid) (PLGA) microparticles (20 or 30 wt%) in rat critical‐sized cranial defects. Materials and methods: Critical‐sized bicortical cranial defects were created in 48 rats and CaP/PLGA cement composites were implanted for 4, 8 and 12 weeks (n=8). Results: Histological analysis of the retrieved specimens revealed that implant degradation was significantly faster for C30% (remaining implant up to 89.4 ± 4.4% at 12 weeks) compared with C20% (remaining implant upto 94.8 ± 2.1% at 12 weeks), albeit that overall degradation was limited. Although bone formation was limited in both experimental groups (upto 685765.9 μm² for C20% vs. 917603.3 μm² for C30%), C30% showed a significant temporal increase of total bone formation. The percentage of defect bridging was comparable for C20% and C30% at all implantation periods (range 40 ± 25.5% at week 4 to 65 ± 20% at week 12 for C20%; range 51.8 ± 7.8% at week 4 to 70.5 ± 16.2% at week 12 for C30%). Conclusion: The amount of PLGA‐microparticles in the CaP/PLGA cement composites demonstrated acceleration of material degradation, while bone formation was found not to be influenced. Further optimization of the composite material is necessary to increase control over degradation and tissue ingrowth. To cite this article :
van de Watering FCJ, van den Beucken JJJP, Walboomers XF, Jansen JA. Calcium phosphate/poly(d ,l ‐lactic‐co‐glycolic acid) composite bone substitute materials: evaluation of temporal degradation and bone ingrowth in a rat critical‐sized cranial defect.
Clin. Oral Impl. Res. 23 , 2012; 151–159.
doi: 10.1111/j.1600‐0501.2011.02218.x     

Evaluation of filling materials in membrane‐protected bone defects. A comparative histomorphometric study in the mandible of miniature pigs.

In recent years, bone grafts and bone substitutes have been increasingly utilized underneath barrier membranes to optimize the treatment outcome of bone reconstructive therapy for defects in the alveolar process. In the present study, 4 different filling materials were evaluated in bone defects of similar dimensions in the mandible of miniature pigs. Blood clots and autografts were used as controls. The defects were covered with barrier membranes and allowed to heal for 4, 12 or 24 weeks. Histologic examination demonstrated that bone repair progressed through a programmed sequence of maturation steps closely resembling the pattern of bone development and growth regardless of whether bone grafts or substitutes were present or not. Histomorphometric analysis showed that autologous bone grafts (autografts) had the best osteoconductive properties during the initial healing period, with 39% of newly formed bone inside the membrane-covered defects at 4 weeks of healing. In addition, 87% of the graft surfaces were already covered by bone at this time. Both values were significantly higher for autografts than for the 4 alternative bone fillers (P < or = 0.05). At 12 weeks, these differences were no longer apparent, with all 5 filling materials showing similar values. Among the tested bone substitutes, tricalcium phosphate (TCP) showed a significantly higher percentage of bone fill at 24 weeks of healing. It can be concluded that sites filled with autografts clearly demonstrated the best results underneath barrier membranes in the early phase of healing. As far as degradation and substitution are concerned, TCP showed the most promising results. This filler, however, needs to be tested further in a more demanding animal model. Less favorable results were obtained for coral-derived hydroxyapatite granules and for demineralized freeze-dried bone allografts.     

骨膜、骨髓单个核细胞复合纳米羟基磷灰石修复骨缺损的实验研究

目的:将骨膜、骨髓单个核细胞(BM-MNCs)、纳米羟基磷灰石(纳米-HA)复合应用于引导性骨再生,为临床修复骨缺损探索一条有效的途径。方法:6-8月龄实验用白兔18只,在兔双侧下颌骨各制备一10 mm×8 mm,深5 mm的骨缺损,按随机数字表法分为3组,每组6只,即A组用自体游离骨膜、骨髓单个核细胞和纳米羟基磷灰石复合修复,B组用自体游离骨膜和纳米羟基磷灰石修复,C组单纯用纳米羟基磷灰石充填修复。分别于术后第4、8、12周末用空气栓塞法处死动物,每次每组处死2只,标本行肉眼、X线、扫描电镜及组织学观察,计算缺损区新骨面积。结果:实验组A组在术后4、8、12周的骨密度值均高于其他两组(P<0.05)。环境扫描电镜观察显示,A组新生骨和原有骨结合致密且骨结构相似。组织学观察表明,A组材料大部分降解,大量成熟骨形成,较少材料包裹于其中,各项指标均优于其他组。结论:骨膜、骨髓单个核细胞、纳米羟基磷灰石复合的成骨作用优秀,可有效促进骨缺损愈合。     

Evaluation of a β‐Calcium Metaphosphate Bone Graft Containing Bone Morphogenetic Protein‐7 in Rabbit Maxillary Defects

Background: Calcium phosphate–based materials have been widely used as bone substitutes and more recently are being exploited together with growth factors as bone tissue engineering scaffolds regulating cell behavior. The aim of this study is to evaluate the in vitro and in vivo response to a newly developed calcium metaphosphate (CMP) bone graft, with and without bone‐stimulating growth factor. Methods: Porous scaffolds of CMP were developed and extensively tested in vitro. Subsequently, CMP grafts with osteogenic protein‐1 (OP‐1) (test) and without OP‐1 (control) were implanted into experimental rabbit maxillary bone defects. Animals were sacrificed at 2, 4, and 8 weeks, and samples were examined with microcomputed tomography (micro‐CT) and processed for histomorphometric analysis. Results: At 8 weeks, the scaffolds containing OP‐1 induced greater bone formation (P = 0.018) than CMP alone, based on histomorphometric evaluation (percentage bone area: test: 57.1 ± 5.6; control: 49.4 ± 7.7) and micro‐CT analysis (percentage bone volume density: test: 63.46 ± 5.61; control: 51.20 ± 6.71). Thus, these data indicated that both test and control CMP grafts showed a good degree of bone formation. Furthermore, the CMP materials showed signs of resorption from 4 weeks, and no graft materials were observed at 8 weeks. Conclusions: In vitro, the OP‐1 loaded graft demonstrated a release profile and bioactivity over a 28‐day period. In vivo testing confirmed enhanced bone formation of the OP‐1 loaded graft after 8 weeks of healing.     

Bone morphogenetic protein-2 enhances bone formation when delivered by a synthetic matrix containing hydroxyapatite/tricalciumphosphate

PURPOSE: The aim of the present study was to test whether or not a synthetic matrix consisting of a polyethylene glycol (PEG) hydrogel containing recombinant human bone morphogenetic protein-2 (rhBMP-2) combined with grafting materials enhances bone regeneration compared with grafting alone or empty control sites. MATERIAL AND METHODS: In each of 10 rabbits, four titanium cylinders were screwed in perforated slits made in the external cortical bones of the calvaria. The following four treatment modalities were randomly allocated: (1) empty control, (2) a combination of a PEG matrix and hydroxyapatite/tricalciumphosphate (HA/TCP) granules and a combination of a PEG matrix containing either 10 microg/ml (3) or 30 microg/ml (4) of BMP-2 and HA/TCP granules. After 8 weeks, the animals were sacrificed and ground sections were obtained for histological analysis. For statistical analysis repeated measures ANOVA and subsequent pairwise Student's t-test were applied (P<0.01). RESULTS: Histomorphometric analysis showed an average area fraction of newly formed bone of 13.96+/-5.98% for the empty control, 15.16+/-7.95% for the PEG and HA/TCP group, 26.32+/-8.56% for the group containing 10 mug rhBMP-2/ml, and 30.15+/-7.63% for the group containing 30 microg rhBMP-2/ml. Statistical analysis revealed significantly more newly formed bone in the two rhBMP-2 groups compared with the PEG and HA/TCP group and with the empty control. Regarding the surface fraction of the HA/TCP graft particles covered with newly formed bone the addition of rhBMP-2 revealed a more than two-fold increase compared with cylinders containing HA/TCP granules without rhBMP-2. This difference reached statistical significance. CONCLUSIONS: It is concluded that rhBMP-2 significantly enhances bone regeneration in rabbits when delivered by a synthetic matrix containing HA/TCP. This synthetic PEG matrix containing HA/TCP granules apparently fulfills a number of criteria required for an ideal carrier system for rhBMP-2.     

Investigation of a Thermoplastic Polymeric Carrier for Bone Tissue Engineering Using Allogeneic Mesenchymal Stem Cells in Granular Scaffolds

PURPOSE: The purpose of this project was to compare alveolar bone repair by allogeneic mesenchymal stem cells using bioglass or synthetic hydroxyapatite (HA)/tricalcium phosphate (TCP) granular scaffolds delivered in a thermoplastic polymeric carrier. MATERIALS AND METHODS: Canine mesenchymal stem cells were obtained from iliac crest bone marrow of beagle dogs and expanded without differentiation. Cells were resuspended at a final concentration of 5 x 10(6) cells/ml in a thermoplastic polymeric carrier (30% w/v Pluronic F-127) and mixed with an equal volume of synthetic HA/TCP or bioglass scaffold and placed into surgically created 5 mm cylindrical defects in the edentulous premolar region of beagle dogs. After 4 weeks or 7 weeks, tissue healing was evaluated by standard histomorphometric methods (Bioquant Nova, Bioquant Image Analysis Corporation, Nashville, TN) by measurement of bone formation within five random sites from each biopsy. RESULTS: After 4 weeks, sites treated with or without mesenchymal stem cells contained 58.25 +/-18.43% or 43.35 +/- 17.68% bone area (p= 0.049), respectively. After 7 weeks, sites treated with or without mesenchymal stem cells contained 62.73 +/- 19.10% or 60.39 +/- 21.32% bone area. Bone formation occurred without inflammation in defects treated using Pluronic F-127 carrier with and without mesenchymal stem cells. There was no difference in percent bone area when bioglass or HA/TCP scaffolds were compared at either time point. CONCLUSIONS: The thermoplastic polymeric carrier did not limit alveolar bone repair in the canine mandible. The combination of a thermoplastic, viscous carrier with a granular scaffold allowed for the delivery of allogeneic mesenchymal stem cells in a clinically manageable form that enhanced bone formation at early stages of alveolar repair.     

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

Objectives: 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. Material and methods: 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. Results: 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. Conclusions: 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.     

Sinus floor augmentation with recombinant human growth and differentiation factor-5 (rhGDF-5): a histological and histomorphometric study in the Goettingen miniature pig

Aim: The aim of this study was to test the hypothesis that recombinant human growth and differentiation factor‐5 (rhGDF‐5) enhances bone formation in sinus floor augmentations in miniature pigs. Material and methods: The maxillary sinus floors in 12 adult female Goettingen minipigs were augmented with β‐tricalcium phosphate (β‐TCP) on one side. The contralateral test side was augmented using two concentrations of rhGDF‐5 (400 μg rhGDF‐5/g β‐TCP; 800 μg rhGDF‐5/g β‐TCP) delivered on β‐TCP (six animals each). One dental implant was inserted into each sinus floor augmentation. After 4 and 12 weeks, histological and histomorphometric assessment of non‐decalcified histological specimens was performed. Results: The results showed significantly higher mean values of volume density (VD) of newly formed bone using the concentration of 400 μg/g β‐TCP (22.8%) compared with the respective control (8%) after 4 weeks (P=0.05). The bone‐to‐implant contact rates were also significantly enhanced after 4 weeks between test sites (400 μg: 41.9%; 800 μg: 40.6%) and control sites (400 μg: 7.8%; 800 μg: 16.4%) (400 μg: P=0.024; 800 μg: P=0.048). Conclusion: It is concluded that rhGDF‐5 delivered on β‐TCP significantly enhanced early bone formation compared with β‐TCP alone in sinus lift procedures in miniature pigs.     

Resorbable and nonresorbable hydroxyapatite granules as bone graft substitutes in rabbit cortical defects

Background: The use of various synthetic calcium phosphate compositions for the promotion of bone in bone defects is of potential interest because such materials may be tailor made and may bond to bone. There is yet an inadequate knowledge of the role of calcium phosphate composition and resorbability for the bone response. Purpose: The aim of the present study was to compare the ability of resorbable versus nonresorbable hydroxyapatite (HA) granules to promote new bone formation in cortical bone defects. Resorbable and nonresorbable HA granules, used as bone graft substitutes, were evaluated after 6 weeks and 3 months in the rabbit tibia. Circular defects (diameter 5.0 mm) were made in both tibias of 18 New Zealand white rabbits. The 36 defects were divided into three groups (six observations per group and time, respectively). The first group was augmented with resorbable HA granules, the second group was augmented with ceramic nonresorbable HA granules, and the third group was left without augmentation (control). The animals were killed after 6 weeks and 3 months, and the tissue was evaluated with light microscopic (LM) morphology and morphometry, scanning electron microscopy (SEM), and energy dispersive x‐ray analysis (EDX). Results: After 3 months LM morphometry revealed significantly more newly formed bone in the two HA augmented groups compared with that in the control. A close contact was found between both kinds of HA granules and new bone as viewed with light microscopy and SEM. A relatively slow degradation process was indicated by the small reduction of the total granule area in the cortical defects. However, LM observations showed a change of granule form. Pilot experiments using SEM‐EDX indicate that Ca and P contents had decreased in the resorbable HA granules between 6 weeks and 3 months. Further, a higher content of Ca and P was found in the newly formed bone close to granules, in comparison with more distant newly formed bone. Conclusions: Our results suggest that both resorbable and nonresorbable HA granules promote new bone formation in rabbit cortical defects, which does not occur in control defects.     

Influence of bone morphogenetic protein and proportion of hydroxyapatite on new bone formation in biphasic calcium phosphate graft: Two pilot studies in animal bony defect model

PurposeThe purpose of these two pilot studies using animal bony defect models was to evaluate the influence of bone morphogenetic protein (BMP) and proportion of hydroxyapatite (HA)/beta-tricalcium phosphate (β-TCP) in biphasic calcium phosphate (BCP) graft on new bone formation.MethodsIn this study, four kinds of synthetic osteoconductive bone materials known for bone growth scaffold, OSTEON™II(HA:β-TCP 30:70), OSTEON™III (HA:β-TCP 20:80), OSTEON™II Collagen, and OSTEON™III Collagen, were prepared as BCP graft materials. In pilot study 1, three BCP materials (OSTEON™II, OSTEON™III, and OSTEON™II Collagen) were grafted in rabbit calvarial defects after impregnating in rhBMP-2. OSTEON™II without the rhBMP-2 impregnation was included in the study as the control. The amount of new bone was examined and measured histologically at 2, 4, and 8 weeks. In pilot study 2, four BCP materials (OSTEON™II, OSTEON™III, OSTEON™II Collagen, and OSTEON™III Collagen) were grafted in beagle dog mandibular defects after soaking in the rhBMP-2. The amount of total bone and new bone were measured three-dimensionally using microCT and healing process was examined histologically at 2, 4, and 8 weeks.ResultsIn pilot study 1, rhBMP-2 impregnated groups showed more new bone formation than the rhBMP-2 free group. In pilot study 2, increased new bone formation was observed in time-dependent manner after graft of BCP and BCP-collagen (OSTEON™II, OSTEON™III, OSTEON™II Collagen, and OSTEON™III Collagen) impregnated with rhBMP-2. Also, BCP with a higher proportion of HA (30% HA) showed more favorable result in new bone formation and space maintenance, especially at the 8 weeks.ConclusionFrom the results of the pilot studies, rhBMP-2 played positive roles in new bone formation and BCP could become a scaffold candidate for rhBMP-2 impregnation to induce new bone formation. Moreover, BCP with a higher proportion of HA (30% HA) could be considered more appropriate for rhBMP-2 carrier.     

Differential effects of bone graft substitutes on regeneration of bone marrow

Background: This study used a rat tibial marrow ablation model to test the hypothesis that bone remodeling within the medullary canal varies with bone graft materials of different chemical compositions and structural properties, impacting marrow cavity restoration. Bone graft materials were selected based on their relative resorption or degradation in vivo and their osteogenic properties. Methods: Following ablation of the right tibial marrow in male Sabra‐strain rats, materials were implanted in the proximal marrow cavity: poly‐d,l ‐lactide‐co‐glycolide 75 : 25 (PLGA); coralline‐hydroxyapatite (HA), calcium‐sulfate (CaSO4), collagen–HA–tricalcium phosphate granules, anorganic bovine bone mineral, demineralized bone matrix (DBM), 45S5 Bioglass (BG), PLGA with BG 50 : 50, PLGA : BG 80 : 20, and PLGA and PLGA:BG 50 : 50 plus bone marrow (BM). Control tibias were ablated but received no implants. At 2 (endosteal bone healing), 4 (marrow cavity remodeling) and 8 weeks (marrow restoration), six to eight animals per group were euthanized and tibias processed for histomorphometry of proximal and distal medullary canals. Results: Control tibias showed primary bone in proximal and distal medullary canals at 2 weeks, with trabeculae surrounded by cellular marrow. At 4 and 8 weeks, control trabeculae were thinned and marrow had more fat cells. In the treated tibias, trabecular bone volume (TBV) varied with time and was material specific. Most implants supported comparable TBV at 2 weeks. Sites with CaSO4 or DBM exhibited decreased TBV with time whereas trabecular bone was retained in proximal tibias containing other materials, closely juxtaposed to the implants. TBV did not always correlate directly with implant volume, but changes in BM volume were correlated inversely with TBV. Addition of BM increased marrow restoration in sites containing PLGA; however, BM reduced restoration of marrow when added to PLGA : BG. Although the presence of implants in the proximal tibia resulted in retention of trabecular bone, there was a time‐dependent reduction in TBV in distal canals; the rate and extent of the distal TBV reduction were implant dependent. Conclusions: Thus, although many materials can support bone formation in the marrow cavity, bone quality, quantity, and physical relationship to the implant, and its rate of resorption differ in a material‐dependent manner, resulting in differences in the restoration of marrow. Clinical relevance: Bone graft materials should be selected not only for their ability to support new bone formation but also for their impact on the remodeling phase of bone healing.     

The bone tissue responses to prehydrated and collagenated cortico-cancellous porcine bone grafts: a study in rabbit maxillary defects

Background: Bone substitutes should have osteoconductive properties and be completely replaced with new bone with time. Adding collagen gel to prehydrated and collagenated porcine bone (PCPB) particles results in a sticky and moldable material which facilitates clinical handling. However, the possible influence of the gel on the bone tissue response is not known. Purpose: The objective of the study was to evaluate the bone tissue responses to PCPB graft with or without collagen gel and to evaluate the resorption/degradation properties of the biomaterials. Materials and Methods: Fourteen rabbits were used in the study. Bilateral bone defects, 5 × 8 × 3 mm, were created in the maxilla and filled with PCPB + collagen gel (test) or with PCPB only (control) and covered with a collagen membrane. Animals were killed after 2 (n = 3), 4 (n = 3), and 8 weeks (n = 8) for histological and morphometrical evaluations. Results: There were no differences between test and control defects. Both materials showed bone formation directly on the particles by typical osteoblastic seams. The bone area increased with time (2–8 weeks) for both sides, from 16.2% (control) and 19.2% (test) to 42.7 and 43.8%, respectively. The PCPB, whether mixed with collagen gel or not, was resorbed by osteoclasts as well as part of remodeling with the formation of osteons within the particles. Morphometry showed a decrease of PCPB area from 19.4% (control) and 23.8% (test) after 2 weeks to 3.7 and 9.3% after 8 weeks, respectively. Conclusions: Mixing collagen gel and PCPB to facilitate the clinical handling does not influence the bone tissue responses to the material, which exhibited osteoconductive properties and was resorbed with time.     

Early effect of platelet-rich plasma on bone healing in combination with an osteoconductive material in rat cranial defects

OBJECTIVE: The early effect of platelet-rich plasma (PRP) on bone regeneration in combination with dense biphasic hydroxyl apatite (HA)/beta-tricalcium phosphate (TCP) particles (ratio 60%/40%) was evaluated in rat cranial defects with a diameter of 6.2 mm. We hypothesize that PRP exerts its beneficial effect on bone regeneration within the first and second week after application in a bone defect combined with an osteoconductive material. MATERIALS AND METHODS: Forty-five rats were used in the study, in which always one cranial defect was created. The defects were filled with HA/beta-TCP particles and HA/beta-TCP particles combined with PRP gel. Some defects were also left unfilled as control. One and two weeks after surgery specimens were retrieved for light microscopy [hematoxylin-eosin, trichrome staining (Masson modification Goldner) and basic fuchsin-methylene blue] and micro-CT analysis to evaluate bone formation and neovascularization. One-way analysis of variance was performed on the raw data obtained from micro-CT analyses. RESULTS: The histological evaluation showed no effect of PRP on bone formation and neovascularization for both implantation times. In the first week, the defect closure was evaluated subjectively to be between 10% and 50% in all samples, whereas no difference among the groups appeared to occur. After 2 weeks, complete bridging of the original bone defect was observed for most of the empty defects, as well as for the defects that contained HA/beta-TCP particles. The trichrome staining revealed no difference in the number of blood vessels between the PRP and non-PRP groups for both implantation times. The osteoconductive nature of dense HA/beta-TCP particles was confirmed, as the bone formation was guided by their outer surfaces and resulted in a larger amount of newly formed bone in comparison with the empty defects. The quantitative micro-CT analysis demonstrated a statistically significant difference in new bone formation between the empty defects and defects filled with particles after 1 week of implantation, but there was no difference between the non-PRP and PRP groups. In at the second week, no difference in bone formation among all groups was observed, whereas even the non-filled control defects were almost completely closed. CONCLUSIONS: A 6.2 mm cranial defect is not a critical-sized defect in rats. Rat PRP had no effect on the early stages of bone healing in addition to an osteoconductive material. Dense HA/beta-TCP particles showed a beneficial effect on bone formation already after 1 and 2 weeks of implantation in non-critical-sized cranial defects in rats.     

Enamel Matrix Derivative,Alone or Associated With a Synthetic Bone Substitute,in the Treatment of 1‐ to 2‐Wall Periodontal Defects

Background: In this study, we compare the effects of enamel matrix derivative (EMD) associated with a hydroxyapatite and β‐tricalcium phosphate (HA/β‐TCP) implant to EMD alone and to open‐flap debridement (OFD) when surgically treating 1‐ to 2‐wall intrabony defects. Methods: Thirty‐four patients, exhibiting ≥3 intraosseous defects in different quadrants, were each treated by OFD, EMD, or EMD + HA/β‐TCP in each defect. At baseline and 12 and 24 months, a complete clinical and radiographic examination was done. Pre‐therapy and post‐therapy clinical (probing depth [PD], clinical attachment level [CAL], and gingival recession [GR]) and radiographic (defect bone level [DBL] and radiographic bone gain [RBG]) parameters for the different treatments were compared. Results: After 12 and 24 months, almost all the clinical and radiographic parameters showed significant changes from baseline within each group (P <0.001). Differences in PD, CAL, and DBL scores were also seen among the three groups at the 12‐ and 24‐month visits (P <0.001). At 12 and 24 months after treatment, the EMD + HA/β‐TCP group showed significantly greater PD reduction (4.00 ± 0.42 mm; 4.25 ± 0.63 mm), CAL gain (3.47 ± 0.65 mm; 3.63 ± 0.91 mm), and RBG (3.17 ± 0.69 mm; 3.35 ± 0.80 mm) and less GR increase (0.56 ± 0.37 mm; 0.63 ± 0.42 mm) compared with the OFD and EMD groups (P <0.05). Conclusion: Our data support the hypothesis that the adjunct of an HA/β‐TCP composite implant with EMD may improve the clinical and radiographic outcomes of the surgical treatment of unfavorable intrabony defects.     

Evaluation of an in situ formed synthetic hydrogel as a biodegradable membrane for guided bone regeneration

The aim of the present study was to test whether or not the application of an in situ formed synthetic hydrogel made of polyethylene glycol (PEG) used as a biodegradable membrane for guided bone regeneration will result in the same amount of bone regeneration as with the use of an expanded polytetrafluoro-ethylene (ePTFE) membrane. In eight New Zealand White rabbits, four evenly distributed 6 mm diameter defects were drilled into the calvarial bone. Three treatment modalities were evenly distributed among the 32 defects: hydroxyapatite (HA)/tricalciumphosphate (TCP) granules covered at the outer and inner surface with a PEG membrane (test), HA/TCP granules covered at the outer and inner surface with an ePTFE membrane (positive control) and HA/TCP granules alone without membranes (negative control). After 4 weeks, the animals were sacrificed and the calvarial bones were removed. The area fraction of newly formed bone was determined by histomorphometrical analysis of the vertical sections from the middle of the defect and by micro-computed tomography of the entire defect. Multiple regression analysis (SAS GLM) was used to model the amount of new bone formation. The quantitative histomorphometric analysis clearly revealed higher values of newly formed bone for the two membrane groups compared with the negative control group. The average area fractions of newly formed bone measured within the former defect amounted to 20.3+/-9.5% for the PEG membrane, 18.9+/-9.9% for the ePTFE membrane, and 7.3+/-5.3% for the sites with no membrane. The micro-computed tomography also showed higher values of new bone formation for the PEG and for the ePTFE groups compared with the negative control group. The GLM revealed a highly significant effect of the treatment on the amount of bone formation (P=0.0048). The values for the negative control group were significantly lower than the ones found in the PEG membrane group (P=0.0017), whereas the ePTFE membrane group showed no significant difference from the PEG membrane group. It is concluded that the PEG membrane can be used successfully as a biodegradable barrier membrane in the treatment of non-critical-size defects in the rabbit skull, and leads to similar amounts of bone regeneration as an ePTFE membrane.     

Healing of rabbit calvarial bone defects using biphasic calcium phosphate ceramics made of submicron‐sized grains with a hierarchical pore structure

Objectives: This study investigated the efficacy of new bone graft substitutes – biphasic calcium phosphates (BCP) made of submicron‐sized grains with fully interconnected wide‐range micron‐scale pores in two different macrodesigns: donut shaped with a 300–400 μm central macropore (n‐BCP‐1) or rod‐shaped (n‐BCP‐2) – in the healing of rabbit calvarial defects, and compared their bone‐healing properties with those of various commercial bone substitutes, which included substitutes with similar BCP composition (MBCP and Osteon), anorganic bovine bone (Bio‐Oss), and β‐TCP (Cerasorb). Material and methods: The surface morphology of the bone substitutes was investigated using scanning electron microscopy (SEM). Defects 8 mm in diameter were created in the calvaria of 30 adult male New Zealand White rabbits and were filled with six types of bone substitutes. The percentage of newly formed bone (NB%) was evaluated histomorphometrically 4 and 8 weeks after implantation. Results: SEM observation showed submicron‐sized grains with fully interconnected micropore structures in the n‐BCP‐1 and n‐BCP‐2 groups; these groups also showed considerable new bone formation in inner micropores as well as on the outer surfaces. The n‐BCP‐1 group exhibited enhanced new bone formation and direct ingrowth of bone tissue with blood vessels into central pores. Histomorphometric analysis showed significantly greater NB% in the n‐BCP‐1 group when compared with the other groups at 4 and 8 weeks (P<0.05). Conclusion: A new BCP ceramics made of submicron‐sized grains with a hierarchical pore structure was an effective osteoconductive material for the treatment of osseous defects of rabbit calvaria. To cite this article:
Park J‐W, Kim E‐S, Jang J‐H, Suh J‐Y, Park K‐B, Hanawa T. Healing of rabbit calvarial bone defects using biphasic calcium phosphate ceramics made of submicron‐sized grains with a hierarchical pore structure.
Clin. Oral Impl. Res. 21 , 2010; 268–276.
doi: 10.1111/j.1600‐0501.2009.01846.x     

The evaluation of processed cancellous bovine bone as a bone graft substitute

OBJECTIVE: To determine the ability of a novel bovine cancellous bone xenoimplant to act as an osteoconductive graft in an ovine femoral defect model. An autograft harvested from the xenoimplant site was placed in a contralateral limb defect for comparison. MATERIAL AND METHODS: The xenoimplant used had been rendered immunologically inert by a novel defatting and deproteinating process. Following surgical implantation of cores into condylar cancellous bone defects, fluorochrome labels were administered to 12 sheep at 2 1/2, 4 1/2 and 8 weeks. Incorporation of the xenoimplants and autografts into the host bone was compared radiographically and histomorphometrically at 10 weeks. RESULTS: Radiographically, the degree of osteointegration was comparable. Histomorphometric data, consisting of labelled surface (LS) estimates, confirmed osteoconductive properties of both the xenoimplants and autografts. Remodelling activity was greatest in the xenoimplants at 2 1/2, weeks. At 4 1/2 weeks, there was more activity in the autograft, but by 8 weeks they were performing similarly. Xenoimplant-LS estimates were comparable or greater than those of the autograft at all times. CONCLUSIONS: Processed bovine cancellous bone xenoimplants were osteoconductive in this model and show promise for development as a biomaterial in human and veterinary orthopaedic surgery.     

The bone regenerative effect of platelet-rich plasma in combination with an osteoconductive material in rat cranial defects

The effect of platelet-rich plasma (PRP) on bone regeneration, in combination with an osteoconductive material, was evaluated in a rat model. Cranial defects, 6.2 mm in diameter, were filled with HA/beta-TCP particles, HA/beta-TCP particles combined with PRP and HA/beta-TCP particles combined with PRP gel, where some were left empty as a control. After 4 weeks of implantation histological, histomorphometrical and micro-computed tomography analyses revealed no difference in new bone formation among the groups. Further, no additional effect of PRP gel in comparison with PRP liquid was detected, except for the increased handling capacity of the graft. These findings suggest that PRP had no positive effect on bone formation in addition to an osteoconductive material after an implantation period of 4 weeks. Also, no negative effect was seen, and neither PRP nor HA/beta-TCP hampered bone ingrowth into the defects.