Use of bovine bone graft and bone membrane in defects surgically created in the cranial vault of rabbits. Histologic comparative analysis

Purpose: This study was proposed to analyze histologically the process of repairing bone defects created surgically in the cranial vaults of rabbits. Materials and Methods: Thirty adult male rabbits (Oryctolagus cunilicus) received, under general anesthesia, bilateral parietal osteotomies by means of a 6mm-diameter trephine. The bony defects were divided into 4 groups. In group 1 the defect did not receive any treatment; in group 2 the defect was filled with lyophilized bovine bone (Biograft); in group 3 it was filled with bovine bone and covered with a bone matrix membrane (Bioplate); in group 4 it was covered with a bone matrix membrane. Animals were sacrificed in 3 equal groups at 15, 30, and 60 days. The specimens were subjected to routine laboratory procedures to evaluate the degree of bone repair. Results: After 60 days, new bone formation in group 2 was not satisfactory when compared to that of group 3. Large amounts of new bone formation in maturation were seen in group 3. In the defects covered with a membrane the results were similar to those of group 1 (ie, the cavity was filled with fibrous connective tissue). The implanted bone and membranes were totally resorbed. Discussion and Conclusions: the use of a membrane served as a barrier against the migration of cells from the adjacent tissue and the bone graft/membrane preserved the cavity space, resulting in an enhanced osteogenic effect.

The influence of local administration of simvastatin in calvarial bone healing in rats

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

Influence of the association between simvastatin and demineralized bovine bone matrix on bone repair in rats

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

Calcitonin in bone-guided regeneration of mandibles in ovariectomized rats: densitometric, histologic and histomorphometric analysis

The aim of the present study was to investigate bone promotion in surgical defects created in the mandible of normal and ovariectomized female rats using calcitonin associated with a polytetrafluoroethylene barrier. The 100 female rats were divided into four groups: control (C), control treated with calcitonin (CM), ovariectomized control (OV) and ovariectomized treated with calcitonin (OVM). A circumscribed bone defect 4 mm in diameter was created in the region of the mandibular angle, and covered with the barrier. Groups CM and OVM received 2 IU/kg of synthetic salmon calcitonin intramuscularly three times a week. The animals were killed 3, 7, 14, 21 and 28 days after surgery. The bone defects were submitted to densitometric, histologic and histomorphometric analysis. Groups C and CM showed higher levels of bone formation after 7 days compared to the OV and OVM groups. A significant difference was observed between groups C and OV at 3-14 days. The OV group presented slower bone regeneration of the surgical bone defect created in the mandibular angle than group C. Synthetic salmon calcitonin accelerated regeneration of the bone defect in the mandibles of OVM animals similarly to group C, and also increased the formation of new bone during the regeneration process in CM.

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

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)

Galea and periosteum flap filled with bone fragments in rabbits

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

Evaluation of digital optical density of bone repair in rats medicated with ketoprofen

O objetivo deste estudo foi avaliar a influência do cetoprofeno sobre o processo de reparação óssea em tíbias de ratos, por meio da análise da densidade óptica digital. Vinte ratos da linhagem Wistar foram divididos em 2 grupos: um grupo controle (sem tratamento) e um grupo tratado com cetoprofeno. Os procedimentos experimentais consistiram de: anestesia, cirurgia, administração do cetoprofeno e exame radiográfico. As imagens radiográficas foram adquiridas empregando-se o sensor digital Visualix GX-S-HDI™ e um aparelho de raios X. As radiografias foram realizadas nos períodos baseline (inicial), 7, 14, 21 e 30 dias pós-operatório, sendo a densidade óptica (DO) avaliada por meio do sistema Vix winTM 1.4. Os valores médios da leitura da do obtidos foram analisados estatisticamente por meio de ANOVA e teste de Tukey com nível de significância de 5%. No grupo controle, houve diferença estatisticamente significante (p=0,001) entre o tempo e a DO, enquanto no grupo tratado com cetoprofeno a diferença não foi estatisticamente significante (p=0,100). O grupo controle apresentou as menores proporções de do (%) no 1º e 7º dias e as maiores proporções de do (%) no 14º, 21º e 30º dias, com diferença estatisticamente significante (p=0,001). Não houve diferença estatisticamente significante (p=0,100) entre as proporções médias de do (%) no grupo tratado, independentemente do período de avaliação. Os achados deste trabalho sugerem que houve influência do cetoprofeno sobre o processo de reparo ósseo, uma vez que na primeira semana o medicamento proporcionou aumento na densidade óptica e provocou atraso na neoformação óssea após o 21º dia.

Evaluation of the tissue response to MTA and MBPC: Microscopic analysis of implants in alveolar bone of rats

The aim of this study was to evaluate and compare the quantitative and qualitative inflammatory responses and bone formation potential after implantation of polyethylene tubes filled with a new calcium hydroxide containing sealer (MBPc) and Prolloot mineral trioxide aggregate (MIA). There were 48 Wistar rats divided in three groups: Group I (control group) empty polyethylene tubes were implanted in the extraction site; group II and III, polyethylene tubes were implanted filled with ProRoot mineral trioxide aggregate (MIA) and MBPc, respectively. At 7, 15, and 30 days after tube implantation, the animals were killed, the hemi-maxillas were removed and prepared to light microscopic analyses. The scores obtained were submitted to Kruskal-Wallis statistical test (p < 0.05). Significant differences between the materials were not observed. The results showed that both materials had similar biological response.

Tibial segmental bone defect treated with bone plate and cage filled with either xenogeneic composite or autologous cortical bone graft

Tibia segmental defect healing in sheep were clinically, radiographically and histologically evaluated. Twelve young sheep aged four to five months were divided into two groups, G1 and G2. A 3.5 cm long segmental defect was created in the right tibial diaphysis with maintenance of the periosteum. The bone defects in both groups were stabilized with a bone plate combined with a titanium cage. In G1 the cage was filled with pieces of autologous cortical bone graft. In G2 it was filled with a composite biomaterial which consisted of inorganic bovine bone, demineralized bovine bone, a pool of bovine bone morphogenetic proteins bound to absorbable ultra-thin powdered hydroxyapatiteand bone-derived denaturized collagen. Except for one G1 animal, all of them showed normal limb function 60 days after surgery. Radiographic examination showed initial formation of periosteal callus in both groups at osteo-tomy sites, over the plate or cage 15 days postoperatively. At 60 and 90 days callus remodeling occurred. Histological and morphometric analysis at 90 days after surgery showed that the quantity of implanted materials in G1 and G2 were similar, and the quantity of new bone formation was less (p = 0.0048) and more immature in G1 than G2, occupying 51 +/- 3.46% and 62 +/- 6.26% of the cage space, respectively. These results suggest that the composite biomaterial tested was a good alternative to autologous cartical bone graft in this experimental ovine tibial defect. However, additional evaluation is warranted prior to its clinical usage.

Bone healing in drill hole defects in spontaneously hypertensive male and female rats' femurs. A histological and histometric study

Objective: the aim of this study was to evaluate the bone healing in spontaneously hypertensive rats (SHR) and compare the results with normotensive rats, evaluating male and female animals.Methods: A bone drill defect was created in the left femur of 24 SHR (12 males and 12 females) and 24 normotensive rats (12 males and 12 females). The animals were divided into two groups and sacrificed 7 and 21 days after the surgical procedure. After the routine laboratory processing, histological and histometric analysis were carried out and data were submitted to ANOVA and Tukey's test (5%).Results: Males and females from the same group had similar histological characteristics. After seven days, all animals presented irregular bone trabeculae. The periosteal osteoblasts were flattened in SHR, and presented a cuboid shape in normotensive animals. After 21 days, the bone defects of all specimens showed a linear closure in all the superficial extension. In addition, SHR presented flattened osteoblasts surrounding the bone trabeculae, while normotensive ones showed cuboidal cells. Statistical analysis of the histometric data indicated similar means between the male and female groups, except for normotensive rats on day 7. In addition, a larger amount of new bone formation was observed in hypertensive when compared to normotensive rats on day 27, in males as well as females.Conclusion: We conclude that bone healing in SHR was more significant than in normotensive ones, as shown by the histological and histometric evaluation 21 days after surgery.

Histomorphometrical analysis of bone formed after maxillary sinus floor augmentation by grafting with a combination of autogenous bone and demineralized freeze-dried bone allograft or hydroxyapatite

Background: Maxillary sinus floor augmentation procedures are currently the treatment of choice when the alveolar crest of the posterior maxilla is insufficient for dental implant anchorage. This procedure aims to obtain enough bone with biomaterial association with the autogenous bone graft to create volume and allow osteo conduction. The objective of this study was to histologically and histometrically evaluate the bone formed after maxillary sinus floor augmentation by grafting with a combination of autogenous bone, from the symphyseal area mixed with DFDBA or hydroxyapatite.Methods: Ten biopsies were taken from 10 patients 10 months after sinus floor augmentation using a combination of 50% autogenous bone plus 50% dernineralized freeze-dried bone allograft (DFDBA group) or 50% autogenous bone plus 50% hydroxyapatite (HA group). Routine histological processing and staining with hernatoxylin and eosin and Masson's trichrome were performed.Results: the histomorphometrical analysis indicated good regenerative results in both groups for the bone tissue mean in the grafted area (50.46 +/- 16.29% for the DFDBA group and 46.79 +/- 8.56% for the HA group). Histological evaluation revealed the presence of mature bone with compact and cancellous areas in both groups. The inflammatory infiltrate was on average nonsignificant and of mononuclear prevalence. Some biopsies showed blocks of the biomaterial in the medullary spaces close to the bone wall, with absence of osteogenic activity.Conclusions: the results indicated that both DFDBA and HA associated with an autogenous bone graft were biocompatible and promoted osteoconduction, acting as a matrix for bone formation. However, both materials were still present after 10 months.

Effect of calcitonin on bone regeneration in male rats: a histomorphometric analysis

The aim of this study was to evaluate the effectiveness of calcitonin in promoting bone growth in surgical bone defects in rat mandibles. Fifty male rats were divided into two groups: bone defect (control) and bone defect with calcitonin (experimental). A circular bone defect 4 mm in diameter was made in the mandibular bone of the rats in the angle region, and covered with a polytetrafluoroethylene barrier. The experimental group received 2 IU/kg of synthetic salmon calcitonin intramuscularly three times a week, with treatment starting immediately after surgery. The animals were killed 3, 7, 14, 21 and 28 days after the surgical procedure. The bone defects were examined histologically and by histomorphometric analysis. The Student t-test was applied to the histomorphometric data, with the level of significance set at 5%. The animals of the experimental group showed a lower level of bone formation at almost all time points than the control group, but no difference between groups was observed 28 days after surgery. The volume of newly formed bone matrix was significantly greater in the control than the experimental group at 7, 14 and 21 days, as determined by both morphologic and histomorphometric analysis. Bone repair in the calcitonin-treated animals was delayed in comparison to controls, indicating the need for further studies on male rats.

Bone healing pattern in surgically created circumferential defects around submerged implants: an experimental study in dog

Objective: To describe the healing of marginal defects below or above 1 mm of dimension around submerged implants in a dog model.Material 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. Two recipient sites were prepared and the marginal 5mm were widened to such an extent to obtain, after implant installation, a marginal gap of 0.5mm at the mesial site (small defect) and of 1.25mm at the distal site (large defect). Titanium healing caps were affixed to the implants and the flaps were sutured allowing a fully submerged healing. The experimental procedures were subsequently performed in the left side of the mandible. The timing of the experiments and sacrifices were planned in such a way to obtain biopsies representing the healing after 5, 10, 20 and 30 days. Ground sections were prepared and histomorphometrically analyzed.Results: The filling of the defect with newly formed bone was incomplete after 1 month of healing in all specimens. Bone formation occurred from the base and the lateral walls of the defects. A larger volume of new bone was formed in the large compared with the small defects. Most of the new bone at the large defect was formed between the 10- and the 20-day period of healing. After 1 month of healing, the outline of the newly formed bone was, however, located at a similar distance from the implant surface (about 0.4mm) at both defect types. Only minor newly formed bone in contact with the implant, starting from the base of the defects, was seen at the large defects (about 0.8mm) while a larger amount was detected at the small defects (about 2.2 mm).Conclusion: Marginal defects around titanium implants appeared to regenerate in 20-30 days by means of a distance osteogenesis. The bone fill of the defects was, however, incomplete after 1 month.

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.