Influence of Osteopenia in Autogenous Bone Graft Healing With or Without Expanded Polytetrafluoroethylene Membranes: Histologic and Histomorphometric Study in Rats

Purpose: The aim of this study was to quantitatively evaluate and qualitatively describe autogenous bone graft healing with or without an expanded polytetrafluoroethylene (e-PTFE) membrane in ovariectornized rats. Materials and Methods: Eighty Wistar rats, weighing approximately 300 g each, were used. A graft was obtained from the parietal bone and fixed to the sidewall of each animal's left mandibular ramus. The animals were randomly divided into four experimental groups (n = 20 in each group): group 1, sham operated and autogenous bone graft only- group 2, sham operated and autogenous bone graft covered by e-PTFE membrane; group 3, ovariectornized (OVX) and autogenous bone graft only- group 4, OVX and autogenous bone graft covered by e-PTFE membrane. The animals were sacrificed at five different time points: immediately after grafting or at 7, 21, 45, or 60 days after grafting. Histologic examination and morphometric measurement of the sections were performed, and values were submitted to statistical analyses. Results: Both groups (sham and OVX) experienced loss of the original graft volume when it was not covered by the membrane, whereas use of the membrane resulted in additional bone formation beyond the edges of the graft and under the membrane. Histologic analysis showed integration of the grafts in all animals, although a larger number of marrow spaces was found in OVX groups. Conclusions: Association of bone graft with an e-PTFE membrane resulted in maintenance of its original volume as well as formation of new bone that filled the space under the membrane. Osteopenia did not influence bone graft repair, regardless of whether or not it was associated with e-PTFE membrane, but descriptive histologic analysis showed larger numbers of marrow spaces in the bone graft and receptor bed and formation of new bone in the OVX animals. INT J ORAL MAXILLOFAC IMPLANTS 2009;24:1074-1082

Vertical Bone Augmentation Using a Polytetrafluoroethylene Nonporous Barrier for Osseointegrated Implants Partially Inserted in Tibiae of Rabbits

Purpose: The aim of this study was to evaluate the possibility of obtaining guided bone regeneration using a poly-tetrafluoroethylene (PTFE) nonporous barrier for 2 endosseous implants, partially inserted in tibiae of rabbits.Materials and Methods: Histologic characteristics of the interface between titanium implants (one group with titanium plasma-coated implants and the other group with acid-treated surfaces) and of the regenerated bone were also studied. Twenty screw-vent implants were placed in tibiae of 5 male New Zealand rabbits, 2 at the right side and 2 at the left side, protruding 3 mm from the bone level, to create a horizontal bone defect. At the experimental group the implants were with a PTFE nonporous barrier, whereas no barriers were used in contralateral implants. Animals were sacrificed 3 months after surgery and biopsy specimens were evaluated histologically and histomorphometrically under light microscopy. Student's t test was used for statistical analysis.Results: The histologic measurements showed a mean gain in bone height of 2.15 and 2.42 mm for the barrier group and 1.95 and 0.43 mm for the control group, for the titanium plasma-spray and acid-treated implant surfaces, respectively.Conclusion: The results of the investigation revealed that the placement of implants protruding 3 nun from crestal bone defects may result in vertical bone augmentation using a nonporous PTFE barrier. (Implant Dent 2009;18:182-191)

Response of human alveolar bone-derived cells to a novel poly(vinylidene fluoride-trifluoroethylene)/barium titanate membrane

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Filling of extraction sockets with autogenous bone in cats

OBJETIVO: Avaliar a regeneração óssea de alvéolos dentais de gatos após enxertia. MÉTODOS: Dezoito gatos adultos, distribuídos em três grupos de 6 animais cada, foram submetidos a extração do canino mandibular direito ou esquerdo. No grupo 1, controle, o alvéolo foi deixado vazio. No grupo 2, o alvéolo foi preenchido com osso esponjoso autógeno do osso ilíaco e no grupo 3, com raspa de osso cortical do osso ilíaco. Os animais foram submetidos à eutanásia 6 semanas após a cirurgia. RESULTADOS: Nas radiografias realizadas no pós-operatório imediato na projeção ventrodorsal observou-se uma área de radiolucência correspondente ao local da alveolectomia e extração dentária. Diminuição da radiolucência foi verificada nas radiografias realizadas seis semanas após a cirurgia. Nos cortes histológicos verificou-se a presença de trabéculas ósseas. A porcentagem de tecido ósseo esponjoso presente nos alvéolos dentais foi quantificada por exames histométricos. Não houve diferença estatisticamente significante entre os grupos (Teste de Kruskal-Wallis p > 0.05) (grupo 1: 52,54 ± 15,4; grupo 2: 50,51 ± 5,01; grupo 3: 51,85 ± 9,52). CONCLUSÃO: Os alvéolos dentais de gatos preenchidos com osso esponjoso autógeno ou raspa de osso cortical autógeno apresentaram regeneração óssea similar àquela observada no grupo controle, após um período de observação de seis semanas.

Evaluation of autogenous bone grafts, particulate or collected during osteotomy with implant burs: Histologic and histomorphometric analysis in rabbits

Purpose: the aim of this study was to evaluate bone regeneration in bone cavities filled with particulate autogenous bone either harvest in blocks and subjected to milling procedures or collected during osteotomy with implant burs. Materials and Methods: In 12 rabbits, 3 noncritical unicortical cavities 7 mm in diameter were prepared with a trephine drill on the right tibia. The cavities were filled respectively with particulate autogenous bone achieved with a manual bone crusher ( particulate group), with particulate autogenous bone obtained using bone collector during osteotomy ( collected group), and with blood clot ( control group). Animals were sacrificed at 7, 15, and 30 days after surgery ( 4 animals for each time period). The sections were examined by histologic and histomorphometric analysis. Results: At 7 days, the samples were filled by coagulum, and bone particles were observed only in the collected (24%) and particulate groups (44.75%). At 15 days, there was connective differentiation in all groups, with presence of grafted bone particles and onset of newly formed bone in the collected (38.88%) and particulate groups (46.0%). At 30 days, there was bone fill ( immature trabecular bone) of the cavities in the control (50%), collected (64.63%) and particulate groups (66%). Conclusion: No significant difference was demonstrated between noncritical unicortical bone defects in rabbit tibiae filled with particulate bone harvested as a block and subjected to milling and those filled with bone collected during osteotomy with implant drills when the defects were observed up to 30 days following their creation.

Implants of polyanionic collagen matrix in bone defects of ovariectomized rats

In recent years, there has been a great interest in the development of biomaterials that could be used in the repair of bone defects. Collagen matrix (CM) has the advantage that it can be modified chemically to improve its mechanical properties. The aim of the present study was to evaluate the effect of three-dimensional membranes of native or anionic (submitted to alkaline treatment for 48 or 96 h) collagen matrix on the consolidation of osteoporosis bone fractures resulting from the gonadal hormone alterations caused by ovariectomy in rats subjected to hormone replacement therapy. The animals received the implants 4 months after ovariectomy and were sacrificed 8 weeks after implantation of the membranes into 4-mm wide bone defects created in the distal third of the femur with a surgical bur. Macroscopic analysis revealed the absence of pathological alterations in the implanted areas, suggesting that the material was biocompatible. Microscopic analysis showed a lower amount of bone ingrowth in the areas receiving the native membrane compared to the bone defects filled with the anionic membranes. In ovariectomized animals receiving anionic membranes, a delay in bone regeneration was observed mainly in animals not subjected to hormone replacement therapy. We conclude that anionic membranes treated with alkaline solution for 48 and 96 h presented better results in terms of bone ingrowth.

Periosteum-Derived Cells as an Alternative to Bone Marrow Cells for Bone Tissue Engineering Around Dental Implants. A Histomorphometric Study in Beagle Dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Deproteinized bovine bone mineral in marginal defects at implants installed immediately into extraction sockets: an experimental study in dogs

Aim: To evaluate the influence of deproteinized bovine bone mineral (DBBM) particles concomitant with the placement of a collagen membrane on alveolar ridge preservation and on osseointegration of implants placed into alveolar sockets immediately after tooth extraction. Material and methods: The pulp tissue of the mesial roots of 3P3 was removed in six Labrador dogs and the root canals were filled. Flaps were elevated in the right side of the mandible, and the buccal and lingual alveolar bony plates were exposed. The third premolar was hemi-sectioned and the distal root was removed. A recipient site was prepared and an implant was placed lingually. After implant installation, defects of about 0.6mm wide and 3.1mm depth resulted at the buccal aspects of the implant, both at the test and at the control sites. The same surgical procedures and measurements were performed on the left side of the mandible. However, DBBM particles with a size of 0.25-1mm were placed into the remaining defect concomitant with the placement of a collagen membrane. Results: All implants were integrated into mature bone. No residual DBBM particles were detected at the test sites after 4 months of healing. Both the test and the control sites showed buccal alveolar bone resorption, 1.8 +/- 1.1 and 2.1 +/- 1mm, respectively. The most coronal bone-to-implant contact at the buccal aspect was 2 +/- 1.1 an 2.8 +/- 1.3mm, at the test and the control sites, respectively. This difference in the distance was statistically significant. Conclusion: The application of DBBM concomitant with a collagen membrane to fill the marginal defects around implants placed into the alveolus immediately after tooth extraction contributed to improved bone regeneration in the defects. However, with regard to buccal bony crest preservation, a limited contribution of DBBM particles was achieved.

Laser 904 nm action on bone repair in rats with osteoporosis

The aim of the present study was to determine the action of AsGA laser irradiation on bone repair in the tibia of osteopenic rats. The animals were randomly divided into eight experimental groups according to the presence of ovarian hormone (sham group) or the absence of the hormone (OVX group), as well as being irradiated or non-irradiated. Low-level 904-nm laser (50 mJ/cm(2)) accelerated the repair process of osteopenic fractures, especially in the initial phase of bone regeneration.Introduction The development of new techniques to speed the process of bone repair has provided significant advances in the treatment of fractures. Some attention recently focused on the effects of biostimulation on bone.Methods Forty-eight adult rats were randomly divided into eight experimental groups (six animals in each group) according to the presence of ovarian hormone (sham group) or absence of the hormone (ovariectomized (OVX) group) as well as being irradiated or non-irradiated. For the application of low-level laser therapy, the animals were anesthetized with one third of the dose sufficient to immobilize the animal and irradiated with AsGa laser (904 nm, 50 mJ/cm(2) for 2s, point form and in contact). The control animals received the same type of manipulation as the irradiated animals, but with the laser turned off. Half of the animals were killed 7 days following the confection of the bone defect, and the other half were killed 21 days after the surgery. After complete demineralization, the tibias were cut cross-sectionally in the central region of the bone defect and embedded in paraffin blocks. The blocks were then cut in semi-seriated slices and stained with hematoxylin and eosin.Results There was new bone formation in the animals in the OVX group with laser treatment killed after 7 days (p<0.001). The lowest percentage of bone formation was observed in the OVX without laser killed after 7 days (p>0.05). All animals killed after 21 days exhibited linear closure of the lesion.Conclusion Low-level 904-nm laser (50 mJ/cm(2)) accelerated the repair process of osteopenic fractures, especially in the initial phase of bone regeneration.

Bone healing in critical-size defects treated with platelet-rich plasma: a histologic and histometric study in rat calvaria

Background and objective: The purpose of this study was to analyze histologically the influence of autologous platelet-rich plasma on bone healing in surgically created critical-size defects in rat calvaria.Material adn Methods: Thirty-two rats were divided into two groups: the control group (group C) and the platelet-rich plasma group. An 8-mm-diameter critical-size defect was created in the calvarium of each animal. In group C the defect was filled by a blood clot only. In the platelet-rich plasma group, 0.35 mL of platelet-rich plasma was placed in the defect and covered by 0.35 mL of platelet-poor plasma. Both groups were divided into subgroups (n = 8) and killed at either 4 or 12 wk postoperatively. Histometric (using image-analysis software) and histologic analyses were performed. The amount of new bone formed was calculated as a percentage of the total area of the original defect. Percentage data were transformed into arccosine for statistical analysis (analysis of variance, Tukey, p < 0.05).Results: No defect completely regenerated with bone. The platelet-rich plasma group had a statistically greater amount of bone formation than group C at both 4 wk (17.68% vs. 7.20%, respectively) and 12 wk (24.69% vs. 11.65%, respectively) postoperatively.Conclusion: Within the limits of this study, it can be concluded that platelet-rich plasma placed in the defects and covered by platelet-poor plasma significantly enhanced bone healing in critical-size defects in rat calvaria.

Bone repair using mineral trioxide aggregate combined to a material carrier, associated or not with calcium hydroxide in bone defects

Mineral trioxide aggregate (MTA) is a powder aggregate containing mineral oxides with a good biological action and may facilitate the regeneration of the periodontal ligament and formation of bone. Calcium hydroxide demonstrates antibacterial properties, enhances tissue dissolution, and induces bone formation. The objective of this study was to evaluate the MTA in the bone healing process and verify if the calcium hydroxide P.A. can improve and accelerate this process. It was used forty rnale Wistar rats, which were divided into two groups, considering or not the use of calcium hydroxide P.A. solution before treatment. Thus, each one of these groups was divided in four groups with five animals each, according to the treatment and the defect filled by: animal's coagulum, monoolein gel, MTA in aqueous solution, and MTA combined with monoolein gel. After 10 days, the animals were perfused and the right hemimandibles removed for histological analysis. Statistical analysis of the data showed significant difference between all analyzed groups when it was made comparisons using or not calcium hydroxide P.A. (p < 0.0001). There was found statistical difference between the groups that was inserted or not MTA, independently the calcium hydroxide application (p < 0.05). Results showed that the MTA used was able to induce bone regeneration and had its action optimized when combined to calcium hydroxide P.A. (C) 2007 Elsevier Ltd. All rights reserved.

Development, characterization and clinical use of a biodegradable composite scaffold for bone engineering in oro-maxillo-facial surgery

We have developed a biodegradable composite scaffold for bone tissue engineering applications with a pore size and interconnecting macroporosity similar to those of human trabecular bone. The scaffold is fabricated by a process of particle leaching and phase inversion from poly(lactide-co-glycolide) (PLGA) and two calcium phosphate (CaP) phases both of which are resorbable by osteoclasts; the first a particulate within the polymer structure and the second a thin ubiquitous coating. The 3-5 mu m thick osteoconductive surface CaP abrogates the putative foreign body giant cell response to the underlying polymer, while the internal CaP phase provides dimensional stability in an otherwise highly compliant structure. The scaffold may be used as a biomaterial alone, as a carrier for cells or a three-phase drug delivery device. Due to the highly interconnected macroporosity ranging from 81% to 91%, with macropores of 0.8 similar to 1.8 mm, and an ability to wick up blood, the scaffold acts as both a clot-retention device and an osteoconductive support for host bone growth. As a cell delivery vehicle, the scaffold can be first seeded with human mesenchymal cells which can then contribute to bone formation in orthotopic implantation sites, as we show in immune-compromised animal hosts. We have also employed this scaffold in both lithomorph and particulate forms in human patients to maintain alveolar bone height following tooth extraction, and augment alveolar bone height through standard sinus lift approaches. We provide a clinical case report of both of these applications; and we show that the scaffold served to regenerate sufficient bone tissue in the wound site to provide a sound foundation for dental implant placement. At the time of writing, such implants have been in occlusal function for periods of up to 3 years in sites regenerated through the use of the scaffold.

Osseous regeneration in the presence of fibrin adhesive material (Tissucol) and epsilon-aminocaproic acid (EACA).

The effects of Tissucol and Tissucol/EACA on bone healing were evaluated histologically. Experimental defects were made in both tibias of 25 rats. Test materials were placed in defects in right tibias and left tibias served as control. Five animals in each group were killed at 1, 3, 7, 14 and 21 days after surgery. Results showed that: a) Tissucol did not interfere with connective and osseous tissue formation; b) Tissucol allowed new bone formation; c) Tissue residues in Tissucol groups in sections of 21-day specimens did not impair healing; d) Tissucol/EACA was usually completely resorbed and healing was complete 21 days after surgery in the Tissucol/EACA group.

A study of different calcification areas in newly formed bone 8 weeks after insertion of dental implants in rabbit tibias

In orofacial implantology there are many types of implants for the different systems. Among these is the implant surface type, e.g., a screw type, cylindrical and laminar. Furthermore, the implants are different in their dimensions, their metal composition, their surface condition, such as smooth, grit or layered surfaces and in their methods of application. Two different self-tapping implants, one smooth and the other grit-blasted, are screwed into the bone, and another one with a plasma of titanium coating, which is also in a screw form but with greater spaces between the screw threads are compared. The greatest amount of bone deposition in the bone/implant interface was encountered in the latter one, the smooth surfaced implant being in second place. All of these systems can alter the implant healing process and to demonstrate this, we injected bone markers in the rabbits over different periods of time so as to observe the different areas of bone deposition in the tibias where the implants had been inserted. The bone tracers used were Alizarin, Calcein and Xylenol-orange. The amount of deposition was calculated by using the method of surface morphometry.

Effects of Chronic Physical Activity and of Ultrasound Treatment on Bone Consolidation

The objective of this study was to investigate the effects of ultrasound treatment and physical exercise on the velocity of bone consolidation and resistance to deformation. We performed osteotomy in the upper third of the right tibia of rats. Physical training consisted of swimming 1 h per day with a load of 5% b.w. for 30 days. Therapy with medium-intensity ultrasound was applied daily on the damaged area. Wistar rats were divided into the following groups: osteotomized sedentary animals with no ultrasound treatment (1.OSnUS), osteotomized trained animals with no ultrasound treatment (2.OTnUS), osteotomized sedentary animals with ultrasound treatment (3.OSwUS). and osteotomized trained animals with ultrasound treatment (4.OTwUS). The animals were sacrificed for the following analyses: muscle glycogen, serum alkaline phosphatase at the 5th, 10th, 20th, and 30th days, test of maximum resistance to flexion, rupture flexion and mean tibial rigidity at the 30th day. Muscle glycogen was increased at the 20th day: alkaline phosphatase was elevated at the 5th and 20th days in groups 3.OSwUS and 4.OTwUS. and decreased at the 10th day. Groups1.OSnUS and 2.OTnUS did not show significant variations. In the mechanical resistance tests, we noted that ultrasound therapy and the association of physical activity used in the present study showed significant differences in bone resistance and bone rigidity after 30 days of treatment. These facts suggest that ultrasound or physical activity, or their combination may accelerate the process of bone tissue repair.