Reconstruction of Mandibular Segmental Defects Using the Guided-Bone Regeneration Technique With Polylactide Membranes and/or Autogenous Bone Graft: A Preliminary Study on the Influence of Membrane Permeability

Purpose: Bone maintenance after mandibular reconstruction with autogenous iliac crest may be disappointing due to extensive resorption in the long term. The potential of the guided-bone regeneration (GBR) technique to enhance the healing process in segmental defects lacks comprehensive scientific documentation. This study aimed to investigate the influence of polylactide membrane permeability on the fate of iliac bone graft (BG) used to treat mandibular segmental defects. Materials and Methods: Unilateral 10-mm-wide segmental defects were created through the mandibles of 34 mongrel dogs. All defects were mechanically stabilized, and the animals were divided into 6 treatment groups: control, BG alone, microporous membrane (poly L/DL-lactide 80/20%) (Mi); Mi plus BG; microporous laser-perforated (15 cm2 ratio) membrane (Mip), and Mip plus BG. Calcein fluorochrome was injected intravenously at 3 months, and animal euthanasia was carried out at 6 months postoperatively. Results: Histomorphometry showed that BG protected by Mip was consistently related to larger amounts of bone compared with other groups (P ≤ .0001). No difference was found between defects treated with Mip alone and BG alone. Mi alone rendered the least bone area and reduced the amount of grafted bone to control levels. Data from bone labeling indicated that the bone formation process was incipient in the BG group at 3 months postoperatively regardless of whether or not it was covered by membrane. In contrast, GBR with Mip tended to enhance bone formation activity at 3 months. Conclusions: The use of Mip alone could be a useful alternative to BG. The combination of Mip membrane and BG efficiently delivered increased bone amounts in segmental defects compared with other treatment modalities. © 2008 American Association of Oral and Maxillofacial Surgeons.

Histological findings following the use of a space-making device for bone reformation and implant integration in the maxillary sinus of primates

Background: Previous studies have shown that membrane elevation results in predictable bone formation in the maxillary sinus provided that implants can be placed as tent poles. In situations with an extremely thin residual crest which impairs implant placement, it is possible that a space-making device can be used under the sinus membrane to promote bone formation prior to placement of implants. Purpose: The present study was conducted to test the hypothesis that the use of a space-making device for elevation of the sinus membrane will result in predictable bone formation at the maxillary sinus floor to allow placement of dental implants. Materials and Methods: Eight tufted capuchin primates underwent bilateral sinus membrane elevation surgery, and a bioresorbable space-making device, about 6 mm wide and 6 mm in height, was placed below the elevated membrane on the sinus floor. An oxidized implant (Nobel Biocare AB, Gothenburg, Sweden) was installed in the residual bone protruding into the created space at one side while the other side was left without an implant. Four animals were sacrificed after 6 months of healing. The remaining four animals received a second implant in the side with a space-making device only and followed for another 3 months before sacrifice. Implant stability was assessed through resonance frequency analysis (RFA) using the Osstell™ (Osstell AB, Gothenburg, Sweden) at installation, 6 months and 9 months after the first surgery. The bone-implant contact (BIC) and bone area inside the threads (BA) were histometrically evaluated in ground sections. Results: Histologically there were only minor or no signs of bone formation in the sites with a space-making device only. Sites with simultaneous implant placement showed bone formation along the implant surface. Sites with delayed implant placement showed minor or no bone formation and/or formation of a dense fibrous tissue along the apical part of the implant surface. In the latter group the apical part of the implant was not covered with the membrane but protruded into the sinus cavity. Conclusions: The use of a space-making device, with the design used in the present study, does not result in bone formation at the sinus floor. However, membrane elevation and simultaneous placement of the device and an implant does result in bone formation at the implant surface while sites with implants placed 6 months after membrane elevation show only small amounts of bone formation. It is suggested that lack of stabilization of the device and/or a too extensive elevation of the membrane may explain the results. © 2009, Wiley Periodicals, Inc.

Evaluation of bone ingrowth into porous titanium implant: Histomorphometric analysis in rabbits

A porous material for bone ingrowth with adequate pore structure and appropriate mechanical properties has long been sought as the ideal bone-implant interface. This study aimed to assess in vivo the influence of three types of porous titanium implant on the new bone ingrowth. The implants were produced by means of a powder metallurgy technique with different porosities and pore sizes: Group 1 = 30% and 180 μm; Group 2 = 30% and 300 μm; and Group 3 = 40% and 180 μm. Six rabbits received one implant of each type in the right and left tibiae and were sacrificed 8 weeks after surgery for histological and histomor-phometric analyses. Histological analysis confirmed new bone in contact with the implant, formed in direction of pores. Histomorphometric evaluation demonstrated that the new bone formation was statistically significantly lower in the group G1 than in group G3, (P = 0.023). Based on these results, increased porosity and pore size were concluded to have a positive effect on the amount of bone ingrowth.

Repair of bone cavities in dog's mandible filled with inorganic bovine bone and bioactive glass associated with platelet rich plasma

The aim of this study was to evaluate the effect of platelet rich plasma (PRP) associated to bovine inorganic bone (Bio-Oss®; Geistlich) or bioactive glass (Bio-Gran®; Orthovita, Implant Innovations) on bone healing. Bone cavities were prepared in both sides of the mandible of 4 adult male dogs. The cavities were divided into 4 groups according to the filling material as follows: control, PRP, PRP/Bio-Oss, PRP/Bio-Gran. The animals were sacrificed after 120 days and histological and histomorphometrical analysis was performed. The control group showed 80.6% of bone formation in the longitudinal sections at 6 mm depth and 83.7% at 13 mm depth. The transverse sections displayed 74.2% at both 6 and 13 mm depths. The PRP group showed 21.1% of bone formation in the longitudinal sections at 6 mm depth, and 23.1% at 13 mm depth. The transverse sections presented 28.98% of bone formation at 6 mm depth and 41.2% at 13 mm depth. The PRP/Bio-Gran group showed 25.1% of bone formation in the longitudinal sections at 6 mm depth and 30.4% at 13 mm depth. In the transverse sections, the bone formation was 43.0% at 6 mm depth and 39.7% at 13 mm depth. The PRP/Bio-Oss group showed 35.5% of bone formation in the longitudinal sections at 6 mm depth and 42% at 13 mm depth. In the transversal sections, the bone formation was 26.8% and 31.2% at the depths of 6 and 13 mm, respectively. PRP alone or associated with bovine inorganic bone or bioglass had no significant effect in bone healing.

Implant osseointegration in circumferential bone defects treated with latex-derived proteins or autogenous bone in dog's mandible

Background: In sites with diminished bone volume, the osseointegration of dental implants can be compromised. Innovative biomaterials have been developed to aid successful osseointegration outcomes. Purpose: The aim of this study was to evaluate the osteogenic potential of angiogenic latex proteins for improved bone formation and osseointegration of dental implants. Materials and Methods: Ten dogs were submitted to bilateral circumferential defects (5.0×6.3mm) in the mandible. Dental implant (3.3×10.0mm, TiUnite MK3™, Nobel Biocare AB, Göteborg, Sweden) was installed in the center of the defects. The gap was filled either with coagulum (Cg), autogenous bone graft (BG), or latex angiogenic proteins pool (LPP). Five animals were sacrificed after 4 weeks and 12 weeks, respectively. Implant stability was evaluated using resonance frequency analysis (Osstell Mentor™, Osstell AB, Göteborg, Sweden), and bone formation was analyzed by histological and histometric analysis. Results: LPP showed bone regeneration similar to BG and Cg at 4 weeks and 12 weeks, respectively (p≥.05). Bone formation, osseointegration, and implant stability improved significantly from 4 to 12 weeks (p≤.05). Conclusion: Based on methodological limitations of this study, Cg alone delivers higher bone formation in the defect as compared with BG at 12 weeks; compared with Cg and BG, the treatment with LPP exhibits no advantage in terms of osteogenic potential in this experimental model, although overall osseointegration was not affected by the treatments employed in this study. © 2009 Wiley Periodicals, Inc.

Biomechanical effect of one session of low-level laser on the bone-titanium implant interface

Low-level laser (LLL) has been used on peri-implant tissues for accelerating bone formation. However, the effect of one session of LLL in the strength of bone-implant interface during early healing process remains unclear. The present study aims to evaluate the removal torque of titanium implants irradiated with LLL during surgical preparation of implant bed, in comparison to non-irradiation. Sixty-four Wistar rats were used. Half of the animals were included in LLL group, while the other half remained as control. All animals had the tibia prepared with a 2 mm drill, and a titanium implant (2.2 × 4 mm) was inserted. Animals from LLL group were irradiated with laser (gallium aluminum arsenide), with a wavelength of 808 nm, a measured power output of 50 mW, to emit radiation in collimated beams (0.4 cm2), for 1 min and 23 s, and an energy density of 11 J/cm2. Two applications (22 J/cm 2) were performed immediately after bed preparation for implant installation. Flaps were sutured, and animals from both groups were sacrificed 7, 15, 30, and 45 days after implant installation, when load necessary for removing implant from bone was evaluated by using a torquimeter. In both groups, torque values tended to increase overtime; and at 30 and 45 days periods, values were statistically higher for LLL group in comparison to control (ANOVA test, p < 0.0001). Thus, it could be suggested that a single session of irradiation with LLL was beneficial to improve bone-implant interface strength, contributing to the osseointegration process. © 2012 Springer-Verlag London Ltd.

Bone-healing pattern at the surface of titanium implants: An experimental study in the dog

Objective: To study the early sequential stages of osseointegration at implants installed in alveolar bony. Materials and methods: In 12 Labrador dogs, all mandibular premolars and first molars were extracted bilaterally. After 3 months of healing, full-thickness flaps were elevated in the edentulous region of the right side of the mandible. Implants were installed, and the flaps were sutured to allow a fully submerged healing. The timing of the installations in the left side of the mandible and of sacrifices were performed with a schedule that various observation periods to sacrifice from 5, 10, 20, and 30 days were available so that n = 6 was obtained per each healing period. Ground sections were prepared and analyzed. Results: Newly formed bone in contact with the implant surface was found after 10 days of healing and the percentage increased up to 50% after 1 month of healing. A higher percentage was found in the trabecular compared with the cortical bony compartment. Old bone decreased by about 50% during healing, being still present after 1 month (16%). The proportions of bone debris and bone particles were at 27% after 5 days and decreased during healing to 6% after 1 month. Conclusion: Osseointegration (new bone-to-implant contact) developed at various rates for cortical and trabecular compartments, respectively. In the trabecular region, mesenchymal cells were identified, subsequently developing into new bone in contact with the implant surface. In the cortical compartment, however, resorptive processes were observed throughout all periods of healing. The proportion of newly formed bone percentage was lower compared with that of the trabecular area. Old bone was still present after 1 month of healing in both compartments. Bone debris and small bone particles appeared to be involved in initial bone formation. © 2013 John Wiley & Sons A/S.

Effects of LLLT in combination with bisphosphonate on bone healing in critical size defects: A histological and histometric study in rat calvaria

The purpose of this study was to analyze histologically the effect of low-level laser therapy (LLLT) in combination with bisphosphonate on bone healing in surgically created critical size defects (CSD) in rat calvaria. One hundred Wistar female rats sham operated (sham) and ovariectomized (Ovx) were maintained untreated for 1 month to allow for the development of osteopenia in the Ovx animals. A CSD was made in the calvarium of each rat, and the animals were divided into five groups according to following treatments: (1) sham rats (control), (2) Ovx rats, (3) Ovx rats treated with LLLT, (4) Ovx rats treated with bisphosphonate, and (5) Ovx rats treated with bisphosphonate and LLLT. Groups 4 and 5 were irrigated with 1 ml of bisphosphonate, and groups 3 and 5 were submitted to LLLT (GaAlAs), 660 nm, 24 J, and 0.4285 W/cm2 on the CSD. Ten animals of each treatment were killed at 30 and 60 days. Histomorphometric assessments, using image analysis software, and histological analyses were performed. No defect was completely regenerated with the bone. Histometrically, it can be observed that groups 3 (37.49 ± 1.94%, 43.11 ± 2.39%) and 5 (35.05 ± 1.57%, 41.07 ± 1.89%) showed a significant bone neoformation when compared to groups 1 (16.81 ± 1.57%, 27.54 ± 1.49%), 2 (11.68 ± 0.98%, 22.51 ± 1.05%), and 4 (14.62 ± 1.70%, 25.67 ± 1.41%) in all experimental periods (P < 0.05). It was possible to conclude that the LLLT associated or not with bisphosphonate treatment was effective for stimulating bone formation in CSD in the calvaria of rats submitted to ovariectomy. © 2012 Springer-Verlag London Ltd.

Bone healing around titanium implants in two rat colitis models

Objective: Crohn's disease is a chronic inflammatory process that has recently been associated with a higher risk of early implant failure. Herein we provide information on the impact of colitis on peri-implant bone formation using preclinical models of chemically induced colitis. Methods: Colitis was induced by intrarectal instillation of 2,4,6-trinitro-benzene-sulfonic-acid (TNBS). Colitis was also induced by feeding rats dextran-sodium-sulfate (DSS) in drinking water. One week after disease induction, titanium miniscrews were inserted into the tibia. Four weeks after implantation, peri-implant bone volume per tissue volume (BV/TV) and bone-to-implant contacts (BIC) were determined by histomorphometric analysis. Results: Cortical histomorphometric parameters were similar in the control (n = 10), DSS (n = 10) and TNBS (n = 8) groups. Cortical BV/TV was 92.2 ± 3.7%, 92.0 ± 3.0% and 92.6 ± 2.7%. Cortical BIC was 81.3 ± 8.8%, 83.2 ± 8.4% and 84.0 ± 7.0%, respectively. No significant differences were observed when comparing the medullary BV/TV and BIC (19.5 ± 6.4%, 16.2 ± 5.6% and 15.4 ± 9.0%) and (48.8 ± 12.9%, 49.2 ± 6.2 and 41.9 ± 11.7%), respectively. Successful induction of colitis was confirmed by loss of body weight and colon morphology. Conclusions: The results suggest bone regeneration around implants is not impaired in chemically induced colitis models. Considering that Crohn's disease can affect any part of the gastrointestinal tract including the mouth, our model only partially reflects the clinical situation. © 2012 John Wiley & Sons A/S.

Bone marrow aspirate combined with low-level laser therapy: A new therapeutic approach to enhance bone healing

This study evaluated the influence of bone marrow aspirate (BMA), low-level laser therapy (LLLT) and their combination on bone healing in surgically created critical-size defects (CSDs) in rat calvaria. 40 rats were divided into four groups: C (control), BMA, LLLT and BMA/LLLT. A 5 mm diameter CSD was created in the calvarium of each animal. In Group C, the defect was filled by blood clot only. In Group BMA, the defect was filled with BMA. In groups LLLT and BMA/LLLT, the defect received laser irradiation (InGaAlP laser), was filled with blood clot or BMA respectively, and irradiated again. Animals were euthanized 30 days postoperatively. Histomorphometric and immunohistochemical analyses were performed. Newly formed bone area (NFBA) was calculated as percentage of the total area of the original defect. Proliferating cell nuclear antigen (PCNA), runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) immunohistochemical staining were performed. PCNA-positive, Runx2-positive and OCN-positive cells were quantified. Data were statistically analyzed. Group BMA/LLLT had significantly greater NFBA than groups C, BMA or LLLT. Group BMA presented significantly greater NFBA than control, while group LLLT did not. Group BMA/LLLT presented a significantly higher number of PCNA-positive and OCN-positive cells than any of the other groups. Groups BMA/LLLT and BMA showed a significantly lower number of Runx2-positive cells than groups C or LLLT. The combination of BMA/LLLT yielded significantly greater bone formation in surgically created CSD in rat calvaria when compared to control, or either treatment alone. © 2013 Elsevier B.V. All rights reserved.

Bone apposition to laminin-1 coated implants: Histologic and 3D evaluation

Laminin-1 has been reported as one of the factors responsible for the nucleation of calcium phosphates and, in vitro, has been reported to selectively recruit osteoprogenitors. This article focused on its in vivo effects, and evaluated the effect of laminin-1 local application on osseointegration. Polished cylindrical hydroxyapatite implants were coated with laminin-1 (test) and the bone responses in the rabbit tibiae after 2 and 4 weeks were evaluated and compared to the non-coated implants (control). Before the samples were processed for histological sectioning, they were three-dimensionally analysed with micro computed tomography (μCT). Both evaluation methods were analysed with regards to bone area around the implant and bone to implant contact. From the histologic observation, new bone formation around the laminin-1 coated implant at 2 weeks seemed to have increased the amount of supporting bone around the implant, however, at 4 weeks, the two groups presented no notable differences. The two-dimensional and three-dimensional morphometric evaluation revealed that both histologic and three-dimensional analysis showed some tendency in favour of the test group implants, however there was no statistical significance between the test and control group results. © 2012 International Association of Oral and Maxillofacial Surgeons.

Increased osseointegration effect of bone morphogenetic protein 2 on dental implants: An in vivo study

Application of recombinant human bone morphogenetic protein 2 (rhBMP-2) to implant surfaces has been of great interest due to its osteoinductive potential. However, the optimal coating methodology has not been clarified. The objective of the study was to determine whether the application of rhBMP-2 onto plasma-sprayed hydroxyapatite implant surfaces by immersion in protein solution before implant installation would result in significantly improved bone apposition. Using a sheep iliac model, titanium (Ti) and plasma-sprayed calcium-phosphate (PSCaP)-coated implants uncoated and coated with rhBMP-2 were assessed for their osteogenic effects in the peri-implant area over time in terms of osseointegration and de novo bone formation. After 3 and 6 weeks postoperatively, the samples were retrieved and were subjected to bone-to-implant contact (BIC) and bone area fraction occupancy (BAFO) evaluation. When rhBMP-2 was applied to the PSCaP surface, significant increases in BIC and BAFO were observed at 3 weeks in vivo, whereas when adsorbed directly onto the titanium implant surface, rhBMP-2 did not as effectively improve the bone response (although significantly higher than control Ti). The outcomes of the present study suggested that the combination of plasma-sprayed calcium-phosphate surface and rhBMP-2 coating significantly enhanced osseointegration, which validated the postulated hypothesis. © 2013 Wiley Periodicals, Inc.

Evaluation of the presence of VEGF, BMP2 and CBFA1 proteins in autogenous bone graft: Histometric and immunohistochemical analysis

Aims: The purpose of this study was to evaluate the expression of proteins that participate in the osteoinduction stage (VEGF, BMP2 and CBFA1) of the process of bone regeneration of defects created in rat calvariae and filled with autogenous bone block grafts. Materials and methods: 10 adult male rats (Rattus norvegicus albinus, Wistar) were used, who received two bone defects measuring 5 mm each in the calvariae. The bone defects constituted two experimental groups (n = 10): Control Group (CONT) (defects filled with a coagulum); Graft Group (GR) (defects filled with autogenous bone removed from the contralateral defect). The animals were submitted to euthanasia at 7 and 30 days post-operatively. Results: Quantitative analysis demonstrated significantly greater bone formation in Group GR, but the presence of the studied proteins was significantly greater in the CONT Group in both time intervals of observation. Conclusion: It was not possible in this study in cortical bone block groups to detect the osteoinductive proteins in a significant amount during the repair process. © 2013 European Association for Cranio-Maxillo-Facial Surgery.

The role of cytokines in inflammatory bone loss

Chronic inflammatory processes close to bone often lead to loss of bone in diseases such as rheumatoid arthritis, periodontitis, loosened joint prosthesis and tooth implants. This is mainly due to local formation of bone resorbing osteoclasts which degrade bone without any subsequent coupling to new bone formation. Crucial for osteoclastogenesis is stimulation of mononuclear osteoclast progenitors by macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-κB ligand (RANKL) which induces their differentiation along the osteoclastic lineage and the fusion to mature, multinucleated osteoclasts. M-CSF and RANKL are produced by osteoblasts/ osteocytes and by synovial and periodontal fibroblasts and the expression is regulated by pro- and anti-inflammatory cytokines. These cytokines also regulate osteoclastic differentiation by direct effects on the progenitor cells. In the present overview, we introduce the basic concepts of osteoclast progenitor cell differentiation and summarize the current knowledge on cytokines stimulating and inhibiting osteoclastogenesis by direct and indirect mechanisms. © Informa Healthcare USA, Inc.

Tomographic follow-up of bone regeneration after bone block harvesting from the mandibular ramus

Autogenous bone is still considered the gold standard, and the applicability of autogenous bone grafts is well established. However, the possibility of second harvesting from the same donor region remains unclear. The aim of this study was to perform a prospective evaluation of hard tissue deposition in the mandibular ramus after bone block harvesting using cone beam computed tomography (CBCT). Twenty-two patients with indications for augmentation procedures using autogenous bone from the mandibular ramus were selected. Three CBCT scans were performed with a tomographic guide before bone harvesting (T1) and at 14 days (T2) and 6 months (T3) after the surgical procedures. Measurements were obtained in 2D (area, mm(2)) and 3D (volume, mm(3)), and were subsequently compared. In the 2D analysis, the mean bone formation rate was 56%, while for the 3D analysis the mean rate was 9.7%. Despite this difference, there was a significant correlation between area and volume measurements. Our findings demonstrated the presence of hard tissue in the mandibular ramus at 6 months after bone harvesting, which suggests that it would be possible to reuse the same region for a second block harvesting. However, the second bone harvesting would involve less bone for transplantation when compared to the first bone harvesting.