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The aim of this study was to evaluate the in vivo performance of two different deproteinized bovine bone (DBB) grafting materials: DBBB (Bio-Oss®) and DBBL (Laddec®), for the regeneration of critically sized (8 mm) defects in rabbit's calvaria. Three round-shaped defects were surgically created in the calvaria of 13 New Zealand White rabbits proximal to the coronal suture in the parietal bone. Two of the defects were filled with one of the grafting materials while a third was left empty to serve as a negative control. Bone regeneration properties were evaluated at 4- and 8-weeks after implantation by means of histological and histomorphometrical analyses. Statistical analyses were performed through a mixed model analysis with fixed factors of time and material. Histological evaluation of the control group evidenced a lack of bridging bone formation across the defect sites at both evaluation time points. For the experimental groups, new bone formation was observed around the defect periphery and to progress radially inwards to the center of the defect site, regardless of the grafting material. Histomorphometric analyses at 4 weeks demonstrated higher amount of bone formation through the defect for DBBB group. However, at 8 weeks, DBBL and DBBB demonstrated osteoconductivity and low resorption rates with evidence of statistically similar bone regeneration through the complete boney defect. Finally, DBBB presented lower soft tissue migration within the defect when compared to DBBL at both evaluation time points. DBBB and DBBL presented similar bone regeneration performance and slow resorption rates. Although both materials promoted bone regeneration through the complete defect, DBBB presented lower soft tissue migration within the defects at 4- and 8-weeks.

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Objectives: This study evaluated the cellular response of primary osteoblasts exposed to two different presentations of a low-temperature non-sintered deproteinized bovine bone matrix (DBBM). Materials and methods: Six different baths of a commercially available DBBM block (Bonefill® Porous Block) and one of DBBM granule (Bonefill® Porous) were evaluated to identify the mineral structure and organic or cellular remnants. Samples of the same baths were processed in TRIZOL for RNA extraction and quantification. For the immunologic cell reaction assay, primary human osteoblasts (pOB) were exposed to DBMM block (pOB + B) or granules (pOB + G), or none (control) for 1, 3, or 7 days of cell cultivation. Expression of proinflammatory cytokines by pOB was evaluated by crosslinked ELISA assay. In addition, total DNA amount, as well as cell viability via LDH evaluation, was assessed. Results: Organic remnants were present in DBBM blocks; 45.55% (±7.12) of osteocytes lacunae presented cellular remnants in blocks compared to 17.31% (±1.31) in granules. In three of five batches of blocks, it was possible to isolate bovine RNA. The highest concentration of TGF-ß1 was found in supernatants of pOB + G on day 7 (218.85 ± 234.62 pg/mL) (p < 0.05), whereas pOB + B presented the lowest amount of TGF-ß1 secretion at the end of evaluation (30.22 ± 14.94 pg/mL, p < 0.05). For IL-6 and OPG, there was no statistical difference between groups, while pOB + G induced more IL-8 secretion than the control (3.03 ± 3.38 ng/mL, p < 0.05). Considering the kinetics of cytokine release during the study period, all groups presented a similar pattern of cytokines, estimated as an increasing concentration for IL-6, IL-8, and OPG during cultivation. Adherent cells were observed on both material surfaces on day 7, according to H&E and OPN staining. Conclusion: Neither tested material induced a pronounced inflammatory response upon osteoblast cultivation. However, further studies are needed to elucidate the potential influence of organic remnants in bone substitute materials on the regeneration process.

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Abstract The limited options for bone repair have led to an extensive research of the field and the development of alloplastic and xenogeneic grafts. The purpose of this study was to evaluate bone repair with two bone substitutes: deproteinized bovine bone (DBB) and biphasic calcium phosphate ceramic (BCP) in critical-size defect. A total of 8-mm defects were made in the parietal bones of rabbits (n=12). The animals were divided into three experimental groups: sham (defect filled with a blood clot), DBB (defect filled with DBB), and BCP (defect filled with BCP). After the experimental periods of 15 and 45 days, the animals were euthanized and submitted to histomorphometric analysis. The total defect area, mineralized tissue area, biomaterial area, and soft tissue area were evaluated. A greater amount of immature bone tissue and biomaterial particles were observed in the BCP group compared to DBB and sham at 45 days (p<0.05). There was no difference in the qualitative pattern of bone deposition between DBB and BCP. However, the sham group did not show osteoid islands along with the defect, presenting a greater amount of collagen fibers as well in relation to the DBB and BCP groups. There was a greater number of inflammatory cells in the DBB at 45 days compared to BCP and sham groups. In conclusion, BCP and DBB are options for optimizing the use of bone grafts for maxillofacial rehabilitation. Bone defects treated with BCP showed greater deposition of bone tissue at 45 days.

Resumo As opções limitadas para reparo ósseo levaram ao desenvolvimento de abrangente pesquisa na área de enxertos aloplásticos e xenogênicos. O objetivo deste estudo foi avaliar o reparo ósseo com dois substitutos ósseos: osso bovino desproteinizado (DBB) e cerâmica fosfática de cálcio bifásica (BCP) em defeito de tamanho crítico. Material e métodos: defeitos críticos de 8 mm foram feitos nos ossos parietais de coelhos (n=12). Os animais foram divididos em três grupos experimentais: sham (defeito preenchido com coágulo sanguíneo), DBB (defeito preenchido com DBB) e BCP (defeito preenchido com BCP). Após os períodos experimentais de 15 e 45 dias, os animais foram sacrificados e submetidos à análise histomorfométrica. Foram avaliadas a área total de defeitos, área de tecidos mineralizados, área de biomateriais e área de tecidos moles. Resultados: maior quantidade de tecido ósseo imaturo e de partículas de biomaterial foram observados no grupo BCP em comparação aos grupos DBB e sham aos 45 dias (p<0,05). Não houve diferença no padrão qualitativo de deposição óssea entre DBB e BCP. Ainda, o grupo sham não apresentou ilhas osteóides ao longo do defeito, apresentando maior quantidade de fibras colágenas em relação aos grupos DBB e BCP. Houve maior quantidade de células inflamatórias no DBB aos 45 dias em comparação aos grupos BCP e sham. Conclusões: BCP e DBB são opções para otimizar o uso de enxertos ósseos na reabilitação de pacientes. Defeitos ósseos tratados com BCP mostraram maior deposição de tecido ósseo aos 45 dias.

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F1-protein fraction (F1) is a natural bioactive compound extracted from the rubber tree, Hevea brasiliensis, and has been recently studied for its therapeutic potential in wound healing. In this study, we investigated the concentration-dependent effects of F1 (0.01%, 0.025%, 0.05%, and 0.1%) incorporated into deproteinized bovine bone (DBB) and porous biphasic calcium phosphate (pBCP), on the repair of rat calvarial critical-size bone defects (CSBD). The defects were analyzed by 3D-microtomography and 2D-histomorphometry at 12 weeks postsurgery. The binding efficiency of F1 to pBCP (96.3 ± 1.4%) was higher than that to DBB (67.7 ± 3.3%). In vivo analysis showed a higher bone volume (BV) gain in all defects treated with DBB (except in 0.1% of F1) and pBCP (except in 0.05% and 0.1% of F1) compared to the CSBD without treatment/control group (9.96 ± 2.8 mm3 ). DBB plus 0.025% F1 promoted the highest BV gain (29.7 ± 2.2 mm3 , p < .0001) compared to DBB without F1 and DBB plus 0.01% and 0.1% of F1. In the pBCP group, incorporation of F1 did not promote bone gain when compared to pBCP without F1 (15.9 ± 4.2 mm3 , p > .05). Additionally, a small BV occurred in defects treated with pBCP plus 0.1% F1 (10.4 ± 1.4 mm3, p < .05). In conclusion, F1 showed a higher bone formation potential in combination with DBB than with pBCP, in a concentration-dependent manner. Incorporation of 0.25% F1 into DBB showed the best results with respect to bone formation/repair in CSBD. These results suggest that DBB plus 0.25% F1 can be used as a promising bioactive material for application in bone tissue engineering.

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OBJECTIVES: The effect of leucocyte- and platelet-rich fibrin (L-PRF), associated with DBBM (deproteinized bovine bone mineral; Bio-Oss®) was investigated and compared with autogenous bone graft as a standard material for filling bone defects. MATERIAL AND METHODS: A defect of 5 mm in diameter was performed in 40 calvaria of rats. The animals were divided into 5 groups and received blood clot (CO), autogenous bone (AUT), DBBM (BIO), L-PRF, or DBBM associated with L-PRF (BIO-LPRF). After 4 and 8 weeks, bone regeneration was assessed by histometric and immunohistochemical analyses. RESULTS: The highest mean percentage of bone formation found at 4 and 8 weeks was observed for the BIO-L-PRF group (54.0% ± 2.8 and 63.6% ± 2.2). The lowest mean percentage at 4 and 8 weeks was observed for the CO group (16.7% ± 2.5 and 20.5% ± 1.0). There was statistical similarity among the AUT, BIO, and L-PRF groups. The expressions OC, RUNX 2, and VEGF showed a favorable aspect in the formation of new bone for BIO-L-PRF. VEGF was the marker with the highest expression because it was related to the initial healing process, promoting the migration and proliferation of endothelial cells in the region of the defect. Even after weeks, VEGF maintained a moderate expression. CONCLUSIONS: The association of L-PRF with DBBM improved bone repair when these biomaterials were inserted into the defects of the calvaria of rats. CLINICAL RELEVANCE: This reinforces the good performance of bovine bone and L-PRF as filler materials, especially when associated.

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BACKGROUND: Bovine bone grafts have been widely used in dentistry for guided tissue regeneration and can support new bone formation in direct contact with the graft. The aim of this study was to compare the morphometric and bone density changes after using two different bovine bone graft blocks in segmental osseous defects in the mandible of rabbits following different postoperative periods. MATERIAL AND METHODS: Critical size segmental defects were surgically created bilaterally in the jaw of 18 rabbits. The defects were filled with either deproteinized bovine bone mineral with 10% collagen (DBBM-C; BioOss Collagen®), lyophilized bovine medullary bone (LBMB; Orthogen®), or left untreated according to a split-mouth design. Animals were sacrificed after 3 or 6 months of healing. The hemimandibles were scanned ex vivo using a high-resolution (19 µm) microcomputed tomography. Morphometric and bone density parameters were calculated in the region of the defect using CT-Analyser (Bruker). Initial graft blocks were used as baseline. RESULTS: DBBM-C presented a denser microarchitecture, in comparison to LBMB at baseline. DBBM-C and LBMB grafted regions showed a similar progressive remodeling, with a significant decrease in structure complexity and maintenance of bone volume fraction during the postoperative follow-up periods. Both graft materials showed an enhanced bone replacement and more complex structure compared to untreated defects. The apparent fusion between the graft and host bone was observed only in the defects filled with LBMB. CONCLUSION: LBMB grafts showed a similar behavior as DBBM-C regarding structural remodeling. In LBMB samples, apparent integration between the host bone and the graft was present.

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The aim of this paper was to evaluate through histological analysis of the tissue reaction of deproteinized bovine bone matrix (DBBM) when inserted into the site of intramuscular ectopic sheep. In this study, 16 sheep received 3 groups and these were divided into 2 experimental times: Group 1-sham group, Group 2-particulate autogenous bone and Group 3-DBBM granules. All animals underwent surgical procedures for insertion of materials in an ectopic site (muscles of the lower back and after 3 and 6 months postoperatively, the samples were evaluated by histological analysis. The results indicated that the Sham group showed dense collagen fibers and thin, characterizing fibrosis at 3 and 6 months. In the autograft group there was a significant amount of collagen deposition and decreased inflammation at 6 months postoperatively. Group of DBBM, it was noted the presence of dense connective tissue and surrounding remaining particles was observed the formation of with osteoid characteristic tissue. The DBBM demonstrated biocompatibility, osteoconductivity and small osteogenesis capacity on ectopic site.

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OBJECTIVE: To compare with pristine sites bone resorption and soft tissue adaptation at implants placed immediately into extraction sockets (IPIES) in conjunction with deproteinized bovine bone mineral (DBBM) particles and a collagen membrane. MATERIAL AND METHODS: The mesial root of the third premolar in the left side of the mandible was endodontically treated (Test). Flaps were elevated, the tooth hemi-sectioned, and the distal root removed to allow the immediate installation of an implant into the extraction socket in a lingual position. DBBM particles were placed into the defect and on the outer contour of the buccal bony ridge, concomitantly with the placement of a collagen membrane. A non-submerged healing was allowed. The premolar on the right side of the mandible was left in situ (control). Ground sections from the center of the implant as well as from the center of the distal root of the third premolar of the opposite side of the mandible were obtained. The histological image from the implant site was superimposed to that of the contralateral pristine distal alveolus, and dimensional variation evaluated for the hard tissue and the alveolar ridge. RESULTS: After 3 months of healing, both histological and photographic evaluation revealed a reduction of hard and soft tissue dimensions. CONCLUSION: The contour augmentation performed with DBBM particles and a collagen membrane at the buccal aspects of implants placed IPIES was not able to maintain the tissue volume.

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OBJECTIVES: To evaluate the influence on osseointegration of Deproteinized bovine bone mineral (DBBM) particles used to fill defects of at least 1 mm around implants having no primary contact with bone. MATERIAL AND METHODS: Premolars and first molars were extracted bilaterally from the mandible of six Labrador dogs. After 3 months of healing, mucoperiosteal full-thickness flaps were elevated, and one recipient site was prepared in the molar region of each hemi-mandible to place implants. These were installed with a deliberate circumferential and periapical space to the bone walls of 1.2 mm. All implants were stabilized with passive fixation plates to maintain the implants in situ and without any contact with the implant bed. The control sites were left to be filled with coagulum, while at the test sites, the residual gap was filled with DBBM. After 3 months of submerged healing, the animals were sacrificed. Ground sections were prepared and analyzed histomorphometrically. RESULTS: Mineralized bone-to-implant contact was 4.0% and 3.9% for control and test sites, respectively. The width of the residual defects was 0.48 mm and 0.88 mm at the control and test sites, respectively. The percentage of implant surface covered by a layer of dense connective tissue of 0.12 mm of width on average was 84.9% and 88.5% at the control and test sites, respectively. CONCLUSION: A minor and not predictable degree of contact or distance osteogenesis was obtained on the implant surface when primary contact of the implant surface with the implant bed had deliberately been avoided. DBBM grafting of the artificial gap did not favor osseointegration. Neither did it enhance the ability to bridge the gap with newly formed bone in an artificial defect wider than 1 mm.

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Objetivo: A proposta deste estudo foi avaliar a regeneração óssea de defeitos criados cirurgicamente e naturais circunferenciais (gap) de implantes pósexodônticos. Materiais e método: Para este estudo foram utilizados 8 cães da raça labrador, nos quais foram instalados implantes imediatos (pós-exodônticos) nos alvéolos dos dentes P2 (2o pré-molar) e M1 (1o molar) inferiores bilaterais. Nos implantes da região dos 2º pré-molares bilaterais foram criados cirurgicamente defeitos peri-implantares vestibulares que foram preenchidos de um lado por osso autógeno – OA e do outro por osso bovino inorgânico Bio-Oss®- OB. Nos alvéolos correspondentes aos primeiros molares (M1) os implantes foram instalados e os gaps (defeitos naturais) existente entre a tábua óssea vestibular e o implante, foram preenchidos por Bone Ceramic® - (grupo BC) ou somente coágulo (grupo COA). Todos os sítios cirúrgicos foram posteriormente recobertos por membrana de colágeno reabsorvível (Biogide ®) e mantidos com cicatrizador. Após 4 meses de pós-operatório, os animais foram eutanasiados e os blocos ósseos processados laboratorialmente para análise histomorfométrica. Resultados: Os resultados obtidos nos implantes instalados na região do P2 mostraram que os valores do IS-C foram ligeiramente maior (2.3±0.8mm) no grupo OA, comparado ao grupo OB (1.7± 0.7 mm), enquanto o IS-B dos grupos OA e OB foram respectivamente de 2.7±0.7mm e 2.2±1.0 mm no lado vestibular e de 1.6±0.8mm e 0.8±0.5mm no lado lingual. As médias dos valores de PM-C, PM-B e PM-IS no grupo OA e OB foram respectivamente de 4.3±0.9mm, 4.7±0.9mm, 2.0±1.6mm e 4.3±0.6mm, 4.8±0.6mm, 2.5±0.8mm. A diferença estatística significativa foi observada somente na espessura da crista óssea vestibular de 2 a 5 mm em direção ao IS (“pescoço do implante”), incluindo o biomaterial (S-OCbt) que se apresentou maior no grupo OB. Nos implantes instalados na região do M1, o gap vestibular horizontal foi de 1.1 (COA) e 1.4 mm (BC) e vertical de 3 e...

Objective: The purpose of these studies were to evaluate bone regeneration at natural or surgically-created buccal defects at implants placed immediately into extraction sockets (IPIES). Material and methods: Eight Labrador dogswere used and implants were placed immediately into the extraction sockets of P2 (second premolar) and M1 (first molar), bilaterally. At P2, the buccal wall of the extraction sockets was removed bilaterally to create a standardized defect, 4 mm wide coronally, 2 mm wide apically, and 6 mm height. At P2 sites, Bio-Oss at the test (OB) or autogenous bone at the control (OA) sites were used to fill the defects. At M1 sites, Bone Ceramic ® at the test (BC) or the clot alone (COA) were used to fill the defects. All surgical sites were subsequently covered by a resorbable collagen membraneand a non-submerged healing was allowed. After 4 months the animals were euthanized, bone blocks harvested and processed for histomorphometric analysis. Results: at P2 sites, IS- C values were slightly higher (2.3±0.8mm) in the OA compared to the OB (1.7±0.7 mm). IS- B were 2.7 ± 0.7mm and 2.2 ± 1.0mm on the buccal side and 1.6 ± 0.8mm and 0.8 ± 0.5mm on the lingual side at OA and OB groups, respectively. The average values of PM- C , B - PM and PM -IS in OA and OB groups were 4.3 ± 0.9mm , 4.7 ± 0.9mm , 2.0 ± 1.6mm and 4.3 ± 0.6mm , 4.8 ± 0.6mm , 2.5 ± 0.8mm, respectively. A statistically significant difference was only observed in the thickness of the buccal bony crest at 2-5 mm apically to IS (the implant neck) when the biomaterial (S -OCbt) was included in the measurement, which appeared to be higher in the OB group. At M1 sites, at the time of installation, the horizontal buccal gap was 1.1 at the COA and 1.4 mm at the BC sites while the vertical remaining defect was 3 mm and 4 mm deep in the control and test sites, respectively. At the buccal aspect, IS-C was higher in the COA (0.6 ± 1.6mm) compared to the BC (0.1 ± 1.8mm) groups and IS- B was...

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Objetivo: A proposta deste estudo foi avaliar a regeneração óssea de defeitos criados cirurgicamente e naturais circunferenciais (gap) de implantes pósexodônticos. Materiais e método: Para este estudo foram utilizados 8 cães da raça labrador, nos quais foram instalados implantes imediatos (pós-exodônticos) nos alvéolos dos dentes P2 (2o pré-molar) e M1 (1o molar) inferiores bilaterais. Nos implantes da região dos 2º pré-molares bilaterais foram criados cirurgicamente defeitos peri-implantares vestibulares que foram preenchidos de um lado por osso autógeno – OA e do outro por osso bovino inorgânico Bio-Oss®- OB. Nos alvéolos correspondentes aos primeiros molares (M1) os implantes foram instalados e os gaps (defeitos naturais) existente entre a tábua óssea vestibular e o implante, foram preenchidos por Bone Ceramic® - (grupo BC) ou somente coágulo (grupo COA). Todos os sítios cirúrgicos foram posteriormente recobertos por membrana de colágeno reabsorvível (Biogide ®) e mantidos com cicatrizador. Após 4 meses de pós-operatório, os animais foram eutanasiados e os blocos ósseos processados laboratorialmente para análise histomorfométrica. Resultados: Os resultados obtidos nos implantes instalados na região do P2 mostraram que os valores do IS-C foram ligeiramente maior (2.3±0.8mm) no grupo OA, comparado ao grupo OB (1.7± 0.7 mm), enquanto o IS-B dos grupos OA e OB foram respectivamente de 2.7±0.7mm e 2.2±1.0 mm no lado vestibular e de 1.6±0.8mm e 0.8±0.5mm no lado lingual. As médias dos valores de PM-C, PM-B e PM-IS no grupo OA e OB foram respectivamente de 4.3±0.9mm, 4.7±0.9mm, 2.0±1.6mm e 4.3±0.6mm, 4.8±0.6mm, 2.5±0.8mm. A diferença estatística significativa foi observada somente na espessura da crista óssea vestibular de 2 a 5 mm em direção ao IS (“pescoço do implante”), incluindo o biomaterial (S-OCbt) que se apresentou maior no grupo OB. Nos implantes instalados na região do M1, o gap vestibular horizontal foi de 1.1 (COA) e 1.4 mm (BC) e vertical de 3 e...

Objective: The purpose of these studies were to evaluate bone regeneration at natural or surgically-created buccal defects at implants placed immediately into extraction sockets (IPIES). Material and methods: Eight Labrador dogswere used and implants were placed immediately into the extraction sockets of P2 (second premolar) and M1 (first molar), bilaterally. At P2, the buccal wall of the extraction sockets was removed bilaterally to create a standardized defect, 4 mm wide coronally, 2 mm wide apically, and 6 mm height. At P2 sites, Bio-Oss at the test (OB) or autogenous bone at the control (OA) sites were used to fill the defects. At M1 sites, Bone Ceramic ® at the test (BC) or the clot alone (COA) were used to fill the defects. All surgical sites were subsequently covered by a resorbable collagen membraneand a non-submerged healing was allowed. After 4 months the animals were euthanized, bone blocks harvested and processed for histomorphometric analysis. Results: at P2 sites, IS- C values were slightly higher (2.3±0.8mm) in the OA compared to the OB (1.7±0.7 mm). IS- B were 2.7 ± 0.7mm and 2.2 ± 1.0mm on the buccal side and 1.6 ± 0.8mm and 0.8 ± 0.5mm on the lingual side at OA and OB groups, respectively. The average values of PM- C , B - PM and PM -IS in OA and OB groups were 4.3 ± 0.9mm , 4.7 ± 0.9mm , 2.0 ± 1.6mm and 4.3 ± 0.6mm , 4.8 ± 0.6mm , 2.5 ± 0.8mm, respectively. A statistically significant difference was only observed in the thickness of the buccal bony crest at 2-5 mm apically to IS (the implant neck) when the biomaterial (S -OCbt) was included in the measurement, which appeared to be higher in the OB group. At M1 sites, at the time of installation, the horizontal buccal gap was 1.1 at the COA and 1.4 mm at the BC sites while the vertical remaining defect was 3 mm and 4 mm deep in the control and test sites, respectively. At the buccal aspect, IS-C was higher in the COA (0.6 ± 1.6mm) compared to the BC (0.1 ± 1.8mm) groups and IS- B was...

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