Gileade Pereira Freitas

Poly(vinylidene-trifluoroethylene)/barium titanate composite for in vivo support of bone formation

In this study, we evaluated the effect of poly(vinylidene fluoride-trifluoroethylene)/barium titanate (P(VDF-TrFE)/BT) membrane on in vivo bone formation. Rat calvarial bone defects were implanted with P(VDF-TrFE)/BT and polytetrafluoroethylene (PTFE) membranes, and at 4 and 8 weeks, histomorphometric and gene expression analyses were performed. A higher amount of bone formation was noticed on P(VDF-TrFE)/BT compared with PTFE. The gene expression of RUNX2, bone sialoprotein, osteocalcin, receptor activator of nuclear factor-kappa B ligand, and osteoprotegerin indicates that P(VDF-TrFE)/BT favored the osteoblast differentiation compared with PTFE. These results evidenced the benefits of using P(VDF-TrFE)/BT to promote new bone formation, which may represent a promising alternative to be employed in guided bone regeneration.

Bone tissue response to plasma-nitrided titanium implant surfaces

A current goal of dental implant research is the development of titanium (Ti) surfaces to improve osseointegration. Plasma nitriding treatments generate surfaces that favor osteoblast differentiation, a key event to the process of osteogenesis. Based on this, it is possible to hypothesize that plasma-nitrided Ti implants may positively impact osseointegration. Objective The aim of this study was to evaluate the in vivo bone response to Ti surfaces modified by plasma-nitriding treatments. Material and Methods Surface treatments consisted of 20% N2 and 80% H2, 450°C and 1.5 mbar during 1 h for planar and 3 h for hollow cathode. Untreated surface was used as control. Ten implants of each surface were placed into rabbit tibiae and 6 weeks post-implantation they were harvested for histological and histomorphometric analyses. Results Bone formation was observed in contact with all implants without statistically significant differences among the evaluated surfaces in terms of bone-to-implant contact, bone area between threads, and bone area within the mirror area. Conclusion Our results indicate that plasma nitriding treatments generate Ti implants that induce similar bone response to the untreated ones. Thus, as these treatments improve the physico-chemical properties of Ti without affecting its biocompatibility, they could be combined with modifications that favor bone formation in order to develop new implant surfaces.

Effect of Surface Nanotopography on Bone Response to Titanium Implant

Clinical success of implant therapy is directly related to titanium (Ti) surface properties and the quality of bone tissue. The treatment of Ti implants with H2SO4/H2O2 is a feasible, reproducible, and low-cost technique to create surface nanotopography (Ti-Nano). As this nanotopography induces osteoblast differentiation, we hypothesized that it may affect bone response to Ti. Thus, this study was designed to evaluate the bone response to a machined Ti implant treated with H2SO4/H2O2 to generate Ti-Nano and to compare it with a commercially available microtopographic Ti implant (Ti-Porous). Implants were placed in rabbit tibias and evaluated after 2 and 6 weeks, and the bone tissue formed around them was assessed by microtomography to record bone volume, bone surface, specific bone surface, trabecular number, trabecular thickness, and trabecular separation. Undecalcified histological sections were used to determine the percentages of bone-to-implant contact, bone area formed between threads, and bone area formed in the mirror area. At the end of 6 weeks, the removal torque was evaluated using a digital torque gauge. The results showed bone formation in close contact with both Ti-Nano and Ti-Porous implants without relevant morphological and morphometric differences, in addition to a similar removal torque irrespective of surface topography. In conclusion, our results have shown that a simple and low-cost method using H2SO4/H2O2 is highly efficient for creating nanotopography on Ti surfaces, which elicits a similar bone response compared with microtopography presented in a commercially available Ti implant.

Selection of reference genes for quantitative real-time polymerase chain reaction studies in rat osteoblasts: ABUNA et al.

Quantitative real‐time polymerase chain reaction (qRT‐PCR) is a powerful tool to evaluate gene expression, but its accuracy depends on the choice and stability of the reference genes used for normalization. In this study, we aimed to identify reference genes for studies on osteoblasts derived from rat bone marrow mesenchymal stem cells (bone marrow osteoblasts), osteoblasts derived from newborn rat calvarial (calvarial osteoblasts), and rat osteosarcoma cell line UMR‐106. The osteoblast phenotype was characterized by ALP activity and extracellular matrix mineralization. Thirty‐one candidates for reference genes from a Taqman® array were assessed by qRT‐PCR, and their expressions were analyzed by five different approaches. The data showed that several of the most traditional reference genes, such as Actb and Gapdh, were inadequate for normalization and that the experimental conditions may affect gene stability. Eif2b1 was frequently identified among the best reference genes in bone marrow osteoblasts, calvarial osteoblasts, and UMR‐106 osteoblasts. Selected stable and unstable reference genes were used to normalize the gene expression of Runx2, Alp, and Oc. The data showed statistically significant differences in the expression of these genes depending on the stability of the reference gene used for normalization, creating a bias that may induce incorrect assumptions in terms of osteoblast characterization of these cells. In conclusion, our study indicates that a rigorous selection of reference genes is a key step in qRT‐PCR studies in osteoblasts to generate precise and reliable data.

Effect of bone morphogenetic protein 9 on osteoblast differentiation of cells grown on titanium with nanotopography

Among bone morphogenetic proteins (BMPs), BMP‐9 has been described as one with higher osteogenic potential. Here, we aimed at evaluating the effect of BMP‐9 on the osteoblast differentiation of cells grown on titanium (Ti) with nanotopography, a well‐known osseoinductive surface. MC3T3‐E1 cells were grown either in absence or presence of BMP‐9 (20 nM) on Ti with nanotopography (Ti‐Nano) or machined Ti (Ti‐Machined) for up to 21 days to evaluate the gene expression of RUNX2, osterix, osteocalcin, bone sialoprotein, SMAD6 and SMAD4, protein expression of SMAD4, ALP activity and extracellular matrix mineralization. As expected BMP‐9 increased osteoblast differentiation irrespective of Ti surface topography; however, the cells grown on Ti‐Nano were more responsible to BMP‐9 compared with cells grown on Ti‐machined. This could be, at least in part, due to the fact that Ti‐Nano may act on both ways, by increasing the activation (SMAD4) and decreasing the inhibition (SMAD6) of the signaling pathway triggered by BMP‐9, while Ti‐Machined only decrease the inhibition (SMAD6) of this pathway. In conclusion, the combination of the osteogenic potential of BMP‐9 with the osseoinductive capacity of Ti‐Nano could be a promising strategy to favor the osseointegration of Ti implants.

Effect of cell therapy with allogeneic osteoblasts on bone repair of rat calvaria defects

BACKGROUND AIMS Regenerative medicine strategies based on cell therapy are considered a promising approach to repair bone defects. The aims of this study were to evaluate the effect of subculturing on the osteogenic potential of osteoblasts derived from newborn rat calvaria and the effect of these osteoblasts on bone repair of rat calvaria defects. METHODS Cells were obtained from 50 newborn rat calvaria, and primary osteoblasts (OB) were compared with first passage (OB-P1) in terms of osteogenic potential by assaying cell proliferation, alkaline phosphatase (ALP) activity, extracellular matrix mineralization and gene expression of the osteoblastic markers RUNX2, ALP, osteocalcin and bone sialoprotein. Then, 5-mm calvaria defects were created in 24 Wistar rats, and after 2 weeks, they were locally injected with 50 µL of phosphate-buffered saline containing either 5 × 106 osteoblasts (OB-P1, n = 12) or no cells (control, n = 12). Four weeks post-injection, the bone formation was evaluated by micro-computed tomography and histological analyses. Data were compared by analysis of variance, followed by the Student-Newman-Keulss test or Students t-test (P ≤ 0.05). RESULTS OB-P1 showed high proliferation and ALP activity, and despite the reduced gene expression of osteoblastic markers and extracellular matrix mineralization compared with OB, they displayed osteogenic potential, being a good choice for injection into calvaria defects. The micro-tomographic and histological data showed that defects treated with OB-P1 presented higher bone formation compared with control defects. DISCUSSION Our results indicate that cells derived from newborn rat calvaria retain osteoblastic characteristics after subculturing and that these osteoblasts stimulate bone repair in a rat calvaria defect model.

Bioactive-glass ceramic with two crystalline phases (BioS-2P) for bone tissue engineering

We aimed to evaluate the in vitro osteogenic and osteoinductive potentials of BioS-2P and its ability to promote in vivo bone repair. To investigate osteogenic potential, UMR-106 osteoblastic cells were cultured on BioS-2P and Bioglass 45S5 discs in osteogenic medium. The osteoinductive potential was evaluated using mesenchymal stem cells (MSCs) cultured on BioS-2P, Bioglass 45S5 and polystyrene in non-osteogenic medium. Rat bone calvarial defects were implanted with BioS-2P scaffolds alone or seeded with MSCs. UMR-106 proliferation was similar for both materials, while alkaline phosphatase (ALP) activity and mineralization were higher for BioS-2P. Bone sialoprotein (BSP), RUNX2 and osteopontin (OPN) gene expression and BSP, OPN, ALP and RUNX2 protein expression were higher on BioS-2P. For MSCs, ALP activity was higher on Bioglass 45S5 than on BioS-2P and was lower on polystyrene. All genes were highly expressed on bioactive glasses compared to polystyrene. BioS-2P scaffolds promoted in vivo bone formation without differences in the morphometric parameters at 4, 8 and 12 weeks. After 8 weeks, the combination of BioS-2P with MSCs did not increase the quantity of new bone compared to the BioS-2P alone. To stimulate osteoblast activity, drive MSC differentiation and promote bone formation, BioS-2P is a good choice as a scaffold for bone tissue engineering.

Surgical Treatment of Fracture in Atrophic Jaw

Introduction:Mandible fractures are common injuries in the facial bones. It has a varied etiology, especially automobile accidents, bicycle accidents, motorcycle accidents and assaults. Objective: This study aims to illustrate a case of a patient with bilateral atrophic mandibular fractures treated surgically. Clinical case:male patient, 48 years old, victim an accident at work causing bilateral mandibular body fractures treated by internal rigid fixation with titanium plates and screws with the making of access submandibular Risdon bilaterally. Conclusion: The authors present a discussion of the controversy involved in this type of treatment and showing the advantages of transcutaneous approach for the treatment of mandibular fractures.