Esther Rieko Takamori

Nanometer Scale Titanium Surface Texturing Are Detected by Signaling Pathways Involving Transient FAK and Src Activations

Background It is known that physico/chemical alterations on biomaterial surfaces have the capability to modulate cellular behavior, affecting early tissue repair. Such surface modifications are aimed to improve early healing response and, clinically, offer the possibility to shorten the time from implant placement to functional loading. Since FAK and Src are intracellular proteins able to predict the quality of osteoblast adhesion, this study evaluated the osteoblast behavior in response to nanometer scale titanium surface texturing by monitoring FAK and Src phosphorylations. Methodology Four engineered titanium surfaces were used for the study: machined (M), dual acid-etched (DAA), resorbable media microblasted and acid-etched (MBAA), and acid-etch microblasted (AAMB). Surfaces were characterized by scanning electron microscopy, interferometry, atomic force microscopy, x-ray photoelectron spectroscopy and energy dispersive X-ray spectroscopy. Thereafter, those 4 samples were used to evaluate their cytotoxicity and interference on FAK and Src phosphorylations. Both Src and FAK were investigated by using specific antibody against specific phosphorylation sites. Principal Findings The results showed that both FAK and Src activations were differently modulated as a function of titanium surfaces physico/chemical configuration and protein adsorption. Conclusions It can be suggested that signaling pathways involving both FAK and Src could provide biomarkers to predict osteoblast adhesion onto different surfaces.

Effect of Roughness of Zirconia and Titanium on Fibroblast Adhesion

The aim of this study was to investigate the adhesion (4 and 24 h) and the morphology of fibroblast Balb/c 3T3 seeded onto polystyrene, partially stabilized (ZrO(2)Y(2)O(3)), stabilized zirconia ceramic (3YTZP), and pure titanium (Ti, grade 2). Initial cell adhesion (4 h) was greater (P < 0.05, analysis of variance and Tukeys Multiple Comparisons Test) onto ZrO(2)Y(2)O(3) and polystyrene than in Ti and 3YTZ. After 24 h, the number of adhered cells was similar between the biomaterials tested, but smaller than onto polystyrene (P < 0.05). Cells were more spread onto glass surface after 4 h, but after 24 h, the morphology and density of the cells were similar in all groups (SEM). Profilometry showed distinct Ra values for each material: glass coverslips and ZrO(2)Y(2)O(3) (0.09 microm), Ti (0.88 microm), and 3YTZP (30.93 microm). It was concluded that ZrO(2)Y(2)O(3) promoted the best initial adhesion, thus indicating that surfaces with Ra values smaller than 0.1 microm could be more favorable to initial adhesion.

Cellular behavior as a dynamic field for exploring bone bioengineering: a closer look at cell-biomaterial interface.

Bone is a highly dynamic and specialized tissue, capable of regenerating itself spontaneously when afflicted by minor injuries. Nevertheless, when major lesions occur, it becomes necessary to use biomaterials, which are not only able to endure the cellular proliferation and migration, but also to substitute the original tissue or integrate itself to it. With the life expectancy growth, regenerative medicine has been gaining constant attention in the reconstructive field of dentistry and orthopedy. Focusing on broadening the therapeutic possibilities for the regeneration of injured organs, the development of biomaterials allied with the applicability of gene therapy and bone bioengineering has been receiving vast attention over the recent years. The progress of cellular and molecular biology techniques gave way to new-guided therapy possibilities. Supported by multidisciplinary activities, tissue engineering combines the interaction of physicists, chemists, biologists, engineers, biotechnologist, dentists and physicians with common goals: the search for materials that could promote and lead cell activity. A well-oriented combining of scaffolds, promoting factors, cells, together with gene therapy advances may open new avenues to bone healing in the near future. In this review, our target was to write a report bringing overall concepts on tissue bioengineering, with a special attention to decisive biological parameters for the development of biomaterials, as well as to discuss known intracellular signal transduction as a new manner to be explored within this field, aiming to predict in vitro the quality of the host cell/material and thus contributing with the development of regenerative medicine.

Evaluation of the cytocompatibility of mixed bovine bone

Treatment of bovine bone with peroxides and chaotropic agents aims to obtain an acellular bone matrix that is able to maintain the collagen-apatite complex and a higher mechanical resistance, a mixed biomaterial hereby named mixed bovine bone (MBB). The purpose of this study was to evaluate the cytocompatibility of MBB and cell-MBB interaction. Cell morphology, number of viable cells, ability to reduce methyltetrazolium and to incorporate neutral red upon exposure to different concentrations of the hydrosoluble extract of MBB were assessed in Balb-c 3T3 cells according to ISO 10993-5 standard. The interaction between cells and MBB surface was evaluated by scanning electron microscopy. The water-soluble MBB extracts were cytotoxic and led to cell death possibly due to its effect on mitochondrial function and membrane permeability. Cells plated directly onto the MBB did not survive, although after dialysis and material conditioning in DMEM + 10% FCS, the cells adhered and proliferated onto the material. It may be concluded that, in vitro, water-soluble MBB extracts were cytotoxic. Nevertheless, MBB cytotoxic effect was reverted by dialysis resulting in a material that is suitable for cell based-therapy in the bioengineering field.

Biocompatibility of Bovine Anorganic Xenograft

Porous granules without organic residues were produced from bovine medular anorganic bone. Sample showed carbonate content and crystalline order similar to natural bone. The cytotoxicity of anorganic bovine xenograft (Ossĕus®) was valuated from fibroblasts (1.5x104/cm2) cultured in serial diluted extract (0 – 100%) of Ossĕus plus 10% of fetal bovine serum (FBS). Pure extract (100%) reduced in 15% the number of viable cells (p<0.05, ANOVA, Tukey test) indicating very low cytotoxicity. Samples were implanted in the subcutaneous tissue of mice in order to evaluate tissue reaction. An organized connective tissue in contact to the granules was observed nine weeks after implantation. The anorganic bovine xenograft (Ossĕus®) was biocompatible and its behavior and osteoconduction potential should be evaluated in bony defects.

Evaluation of UCLA Implant-Abutment Sealing

PURPOSE To evaluate the effect of the presence of a prefabricated cobalt-chromium (CoCr) margin in a universal castable long abutment (UCLA) on the sealing capability and fit of the implant-abutment interface. MATERIALS AND METHODS One-hundred twenty external hexagon implants (SIN) were divided into two groups (n=60 each) to receive UCLA abutments from six manufacturers (n=10 each) either with or without a CoCr margin (n=60 each). Abutments were cast and 12 groups were formed: M (Microplant), I (Impladen), S (SIN), Sv (Signo Vinces), T (TitaniumFix), and B (Bionnovation). Sealing was determined by placing 0.7 µL of 0.1% toluidine blue in the implant wells before abutment torquing. Implant-abutment samples were placed into 2.0-mL vials containing 0.7 mL of distilled water to maintain the implant-abutment interface, and aliquots of 100 µL of water were retrieved at 1, 3, 6, 24, 48, 72, 96, and 144 hour incubation times for measurement of absorbance in a spectrophotometer, and returned for repeated measurements. Two-way ANOVA (P<.05) and Tukeys test were used. Scanning electron microscopy (SEM) was used for observation of the implant-abutment fit. RESULTS Groups M, Sv, and T without the CoCr margin resulted in complete release of toluidine blue at 1 hour, whereas I, S, and B did so at 3, 24, and 96 hours, respectively. Complete leakage in abutments with the prefabricated margin occurred at 6 hours for S; 24 hours for Sv, T, and B; and 72 hours for M and I. Implant-abutment gaps were observed in all groups. A poorer fit was depicted for groups M and T without the CoCr margin. CONCLUSION Complete leakage was observed for all UCLA abutments regardless of the presence of the CoCr margin. Implant-abutment gaps were observed in all groups.

Porcine peritoneum as source of biocompatible collagen in mice

PURPOSE: To investigate the biocompatibility and biodegradability of a membrane made from porcine peritoneum. METHODS: The membrane (5x5 mm) was inserted in the subcutaneous tissue on the back of 15 mice, which were killed after 1, 3 and 9 weeks (ISO 10993-6). The cellular components of the inflammatory response and degradation of the membrane were analyzed in hematoxylin-eosin-stained histological sections. RESULTS: After one week, mononuclear cells were observed inside the membrane. After three weeks, the material was almost completely absorbed. After nine weeks, there was no presence of material and there were signs of tissue remodeling. There was neither a foreign body reaction nor signs of tissue necrosis. CONCLUSION: The collagen membrane derived from porcine peritoneum is biocompatible and bioabsorbable when implanted in the subcutaneous tissue of mice.