Mesenchymal Stem Cells and Bovine Bone Mineral in Sinus Lift Procedures-An Experimental Study in Sheep

Introduction: New reconstructive and less invasive methods have been searched to optimize bone formation and osseointegration of dental implants in maxillary sinus augmentation. Purpose: The aim of the presented ovine split-mouth study was to compare bovine bone mineral (BBM) alone and in combination with mesenchymal stem cells (MSCs) regarding their potential in sinus augmentation. Material and Methods: Bilateral sinus floor augmentations were performed in six adult sheep. BBM and MSCs were placed into the test side and only BBM in the contra-lateral control side of each sheep. Animals were sacrificed after 8 and 16 weeks. Augmentation sites were analyzed by computed tomography, histology, and histomorphometry. Results: The initial volumes of both sides were similar and did not change significantly with time. A tight connection between the particles of BBM and the new bone was observed histologically. Bone formation was significantly (p = 0.027) faster by 49% in the test sides. Conclusion: The combination of BBM and MSCs accelerated new bone formation in this model of maxillary sinus augmentation. This could allow early placement of implants.

Mesenchymal stem cells and inorganic bovine bone mineral in sinus augmentation: comparison with augmentation by autologous bone in adult sheep

Our aim was to compare the osteogenic potential of mononuclear cells harvested from the iliac crest combined with bovine bone mineral (BBM) (experimental group) with that of autogenous cancellous bone alone (control group). We studied bilateral augmentations of the sinus floor in 6 adult sheep. BBM and mononuclear cells (MNC) were mixed and placed into one side and autogenous bone in the other side. Animals were killed after 8 and 16 weeks. Sites of augmentation were analysed radiographically and histologically. The mean (SD) augmentation volume was 3.0 (1.0) cm(3) and 2.7 (0.3) cm(3) after 8 and 16 weeks in the test group, and 2.8 (0.3) cm(3) (8 weeks) and 2.8 (1.2) cm(3) (16 weeks) in the control group, respectively. After 8 weeks, histomorphometric analysis showed 24 (3)% BBM, and 19 (11)% of newly formed bone in the test group. The control group had 20 (13%) of newly formed bone. Specimens after 16 weeks showed 29 (12%) of newly formed bone and 19 (3%) BBM in the test group. The amount of newly formed bone in the control group was 16 (6%). The results show that mononuclear cells, including mesenchymal stem cells, in combination with BBM as the biomaterial, have the potential to form bone. (C) 2009 The British Association of Oral and Maxillofacial Surgeons. Published by Elsevier Ltd. All rights reserved.

A Clinical Study of 406 Sinus Augmentations With 100% Anorganic Bovine Bone

Background: The aim of the present study is to evaluate the use of anorganic bovine bone (ABB) associated with a collagen membrane (CM) for a sinus graft by means of clinical, histologic, and radiographic parameters in cases with bone availability <= 7 mm. A preliminary evaluation consisted of a clinical examination, computed tomography (CT), and a panoramic x-ray. Methods: Ninety-two patients requiring bilateral sinus grafts and 222 requiring unilateral procedures (total: 406 sinuses) participated in this study. A total of 1,025 implants were placed in the grafted sinuses. A total of 118 implants were placed simultaneously with the sinus graft (one stage), and 907 implants were placed in a subsequent surgery (two stages), 6 to 12 months after the graft was performed. In seven cases, a biopsy was harvested for histomorphometric analysis. Recall appointments were scheduled every 6 months, and panoramic and periapical x-rays were required every year for 3 years. Results: Among 1,025 implants, 19 were lost (survival rate: 98.1%). The difference in survival rates for implants placed in native bone: <= 3 mm (98.1%), >3 to <= 5 mm (98.6%), and >5 to <= 7 mm (97.0%) was not statistically significant (P = 0.3408). The survival rates for implants with rough and machined surfaces (98.6% and 97.0%, respectively) were not statistically significant (P = 0.0840). The histomorphometric analysis showed new bone formation (39.0% +/- 12%), marrow space (52.9% +/- 9.3%), and residual ABB (8% +/- 2.7%). Conclusion: Our results indicated that 1,025 implants placed in sinuses grafted exclusively with ABB combined with CM led to an excellent and predictable survival rate of 98.1%. J Periodontol 2009;80:1920-1927.

Elastic modulus and hardness of cortical and trabecular bovine bone measured by nanoindentation

The elastic modulus and hardness of several microstructure components of dry bovine vertebrae and tibia have been investigated in the longitude and transverse directions using nanoindentation. The elastic modulus for the osteons and the interstitial lamellae in the longitude direction were found to be (24.7±2.5) GPa and (30.1±2.4) GPa. As it's difficult to distinguish osteons from interstitial lamellae in the transverse direction, the average elastic modulus for cortical bovine bone in the transverse direction was (19.8±1.6) GPa. The elastic modulus for trabecular bone in the longitude and transverse direction were (20±2) GPa and (14.7±1.9) GPa respectively. The hardness also varied among the microstructure components in the range of 0.41–0.89 GPa. Analyses of variance show that the values are significantly different.

Nano- and macro-scale characterisation of the mechanical properties of bovine bone

In the present study, nano- and macro-scale characterisations on the mechanical properties of bovine cortical bones have been performed by using nanoindentation and conventional compressive tests. Nanoindentation results showed that the elastic modulus for the osteons and the interstitial lamellae in the longitude direction were 24.7 ± 2.5 GPa and 30.1 ± 2.4 GPa. As it’s difficult to distinguish osteons from interstitial lamellae in the transverse direction, the average elastic modulus for cortical bovine bone in the transverse direction was 19.8 ± 1.6 GPa. Significant differences were found in the modulus values between different microstructures of bone tissue and in different testing direction. It was found that the elastic modulus of bone bovine material in nano-level was higher than that in macro-level. The elastic modulus and
ultimate stress of large bone samples were 12.5 ± 1.9 GPa and 195 ± 19 MPa respectively from the compression test.

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.

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)

Effect of bovine bone morphogenetic proteins on radius fracture healing in rabbits

OBJETIVO: Investigar a influência de Proteínas Morfogenéticas Ósseas de origem bovina (bBMPs) ligadas a hidroxiapatita mais colágeno na consolidação de fraturas instáveis do rádio. MÉTODOS: em 15 coelhos com aproximadamente 5,5 meses de idade e peso médio de 3,5kg foi realizada uma fratura transversa na porção média da diáfise do rádio de ambos os membros. Na fratura do rádio direito foi aplicada mistura de bBMPs ligadas à hidroxiapatita (bBMP-HA) e colágeno bovino como aglutinante e na do rádio esquerdo, considerada controle, nenhum tratamento foi usado. Os coelhos (cinco por período) foram submetidos à eutanásia aos 30, 60 e 90 dias após a cirurgia para realização do processamento histológico e análise microscópica. RESULTADOS: A análise histológica descritiva revelou que a consolidação foi similar para os membros tratado e controle. Pela análise histomorfométrica, a área de novo osso foi em média 867442,16 mm², 938743.00 mm² e 779621,06 mm² para os membros controles e 841118,47 mm², 788038,76mm² e 618587,24 mm² para os membros tratados, aos 30, 60 e 90 dias, respectivamente. Desta forma, aos 60 dias de pós-operatório a área de novo osso foi 12.17% maior no membro tratado com bBMP-HA/colágeno em relação ao membro controle (p<0.05, teste de Tukey). em ambos os membros a área de novo osso aumentou durante o período experimental até a total consolidação da fratura. CONCLUSÃO: Baseado nos resultados obtidos foi possível concluir que a mistura de bBMP-HA/colágeno induziu pequena, porém significante melhora na consolidação da fratura.

Alveolar process preservation at implants installed immediately into extraction sockets using deproteinized bovine bone mineral - an experimental study in dogs

Aim To evaluate the soft tissue and the dimensional changes of the alveolar bony crest at sites where deproteinized bovine bone mineral (DBBM) particles, concomitantly with the placement of a collagen membrane, were used at implants installed into 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 bilaterally, the premolars hemi-sectioned, and the distal roots removed. Recipient sites were prepared in the distal alveolus, and implants were placed. At the test sites, DBBM particles were placed in the residual marginal defects concomitantly with the placement of a collagen membrane. No treatment augmentation was performed at the control sites. A non-submerged healing was allowed. Impressions were obtained at baseline and at the time of sacrifice performed 4 months after surgery. The cast models obtained were analyzed using an optical system to evaluate dimensional variations. Block sections of the implant sites were obtained for histological processing and soft tissue assessments. Results After 4 months of healing, no differences in soft tissue dimensions were found between the test and control sites based on the histological assessments. The location of the soft tissue at the buccal aspect was, however, more coronal at the test compared with the control sites (1.8 +/- 0.8 and 0.9 +/- 0.8 mm, respectively). At the three-dimensional evaluation, the margin of the soft tissues at the buccal aspect appeared to be located more apically and lingually. The vertical dislocation was 1 +/- 0.6 and 2.7 +/- 0.5 mm at the test and control sites, respectively. The area of the buccal shrinkage of the alveolar crest was significantly smaller at the test sites (5.9 +/- 2.4 mm2) compared with the control sites (11.5 +/- 1.7 mm2). Conclusion The use of DBBM particles concomitantly with the application of a collagen membrane used at implants placed into sockets immediately after tooth extraction contributed to the preservation of the alveolar process.

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.

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.

Dispersed hydroxyapatite bioglass 45S5 composites: Comparative evaluation of the use of bovine bone and synthetic hydroxyapatite

The bovine bone and sintetic hydroxyapatite (HA) bioceramics are reference materials to employment as a bone substitute, however, their slow rate of degradation and its low rate of bioactivity index (Ib) are presented as limiting factors for application as bone graft. In contrast, the bioglass is a resorbable and osteoinductive material. the present work objective the development of composites of dispersed bovine bone or sintetic HA in silicate-phosphate bioglass, seeking to obtain a biomaterial with properties suitable for application as bone grafts. The composites were prepared by mixing between the powder components followed by sintering for 1h. Were used HA and bioglass (45S5) with particle size <240μm. The tested proportions of HA/45S5 were 20/80, 30/70 and 40/60 (wt%). The composites characterization was made employing scanning electron microscopy, Infra-Red Spectrometry and hydrolytic resistance test. The test results indicate the potential use of the materials developed for applications such as bone graft. © (2012) Trans Tech Publications, Switzerland.

Autogenous bone combined with anorganic bovine bone for maxillary sinus augmentation: Analysis of the osteogenic potential of cells derived from the donor and the grafted sites

Objectives: This study aimed to comparatively evaluate the in vitro osteogenic potential of cells obtained from the mandibular ramus (MR, autogenous bone donor site) and from the maxillary sinus (MS) bone grafted with a mixture of anorganic bovine bone (ABB) and MR prior to titanium implant placement (MS, grafted implant site). Material and methods: Cells were obtained from three patients subjected to MS floor augmentation with a 1: 1 mixture of ABB (GenOx Inorg®) and MR. At the time of the sinus lift procedure and after 8 months, prior to implant placement, bone fragments were taken from MR and MS, respectively, and subjected to trypsin-collagenase digestion for primary cell culturing. Subcultured cells were grown under osteogenic condition for up to 21 days and assayed for proliferation/viability, osteoblast marker mRNA levels, alkaline phosphatase (ALP) activity and calcium content/Alizarin red staining. ALP activity was also determined in primary explant cultures exposed to GenOx Inorg® (1: 1 with MR) for 7 days. Data were compared using either the Mann-Whitney U-test or the Kruskal-Wallis test. Results: MS cultures exhibited a significantly lower osteogenic potential compared with MR cultures, with a progressive increase in cell proliferation together with a decrease in osteoblast markers, reduced ALP activity and calcium content. Exposure of MR-derived primary cultures to GenOx Inorg® inhibited ALP activity. Conclusion: These results suggest that the use of GenOx Inorg® in combination with MR fragments for MS floor augmentation inhibits the osteoblast cell differentiation at the implant site in the long term. © 2013 John Wiley & Sons A/S.

Deproteinized bovine bone mineral particles and osseointegration of implants without primary bone contact: An experimental study in dogs

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. © 2013 John Wiley & Sons A/S.