Adalberto L Rosa

Publications of Adalberto L Rosa

• Effects of enamel matrix derivative and transforming growth factor-β1 on human osteoblastic cells.

  Authors: Daniela B Palioto, Thaisângela L Rodrigues, Julie T Marchesan, Márcio M Beloti, Paulo T de Oliveira, Adalberto L Rosa

  Head & face medicine. 07/2011; 7:13.

  Extracellular matrix proteins are key factors that influence the regenerative capacity of tissues. The objective of the present study was to evaluate the effects of enamel matrix derivative (EMD),Extracellular matrix proteins are key factors that influence the regenerative capacity of tissues. The objective of the present study was to evaluate the effects of enamel matrix derivative (EMD), TGF-β1, and the combination of both factors (EMD+TGF-β1) on human osteoblastic cell cultures. Cells were obtained from alveolar bone of three adult patients using enzymatic digestion. Effects of EMD, TGF-β1, or a combination of both were analyzed on cell proliferation, bone sialoprotein (BSP), osteopontin (OPN) and alkaline phosphatase (ALP) immunodetection, total protein synthesis, ALP activity and bone-like nodule formation. All treatments significantly increased cell proliferation compared to the control group at 24 h and 4 days. At day 7, EMD group showed higher cell proliferation compared to TGF-β1, EMD + TGF-β1 and the control group. OPN was detected in the majority of the cells for all groups, whereas fluorescence intensities for ALP labeling were greater in the control than in treated groups; BSP was not detected in all groups. All treatments decreased ALP levels at 7 and 14 days and bone-like nodule formation at 21 days compared to the control group. The exposure of human osteoblastic cells to EMD, TGF-β1 and the combination of factors in vitro supports the development of a less differentiated phenotype, with enhanced proliferative activity and total cell number, and reduced ALP activity levels and matrix mineralization.

• Efficacy of a bioactive glass-ceramic (Biosilicate((R))) in the maintenance of alveolar ridges and in osseointegration of titanium implants.

  Authors: Virgílio M Roriz, Adalberto L Rosa, Oscar Peitl, Edgar D Zanotto, Heitor Panzeri, Paulo T de Oliveira

  Clinical oral implants research. 11/2009;

  Abstract Objectives: The aims of this research were to evaluate the efficacy of a bioactive glass-ceramic (Biosilicate((R))) and a bioactive glass (Biogran((R))) placed in dental sockets in theAbstract Objectives: The aims of this research were to evaluate the efficacy of a bioactive glass-ceramic (Biosilicate((R))) and a bioactive glass (Biogran((R))) placed in dental sockets in the maintenance of alveolar ridge and in the osseointegration of Ti implants. Material and methods: Six dogs had their low premolars extracted and the sockets were implanted with Biosilicate((R)), Biogran((R)) particles, or left untreated. After the extractions, measurements of width and height on the alveolar ridge were taken. After 12 weeks a new surgery was performed to take the final ridge measurements and to insert bilaterally three Ti implants in biomaterial-implanted and control sites. Eight weeks post-Ti implant placement block biopsies were processed for histological and histomorphometric analysis. The percentages of bone-implant contact (BIC), of mineralized bone area between threads (BABT), and of mineralized bone area within the mirror area (BAMA) were determined. Results: The presence of Biosilicate((R)) or Biogran((R)) particles preserved alveolar ridge height without affecting its width. No significant differences in terms of BIC, BAMA, and BABT values were detected among Biosilicate((R)), Biogran((R)), and the non-implanted group. Conclusions: The results of the present study indicate that filling of sockets with either Biosilicate((R)) or Biogran((R)) particles preserves alveolar bone ridge height and allows osseointegration of Ti implants. To cite this article: Roriz VM, Rosa AL, Peitl O, Zanotto ED, Panzeri H, de Oliveira PT. Efficacy of a bioactive glass-ceramic (Biosilicate((R))) in the maintenance of alveolar ridges and in osseointegration of titanium implants. Clin. Oral Impl. Res. xx, 2009; 000-000.

• In vitro proliferation and osteoblastic phenotype expression of cells derived from human vertebral lamina and iliac crest.

  Authors: Helton L A Defino, Carlos F P da Silva Herrero, Grasiele E Crippa, Larissa Sverzut Bellesini, Marcio M Beloti, Adalberto L Rosa

  Spine. 08/2009; 34(15):1549-53.

  STUDY DESIGN: Osteoblastic cells derived from vertebral lamina and iliac crest were isolated and cultured under the same conditions (osteogenic medium, pH, temperature, and CO2 levels). OBJECTIVE: ToSTUDY DESIGN: Osteoblastic cells derived from vertebral lamina and iliac crest were isolated and cultured under the same conditions (osteogenic medium, pH, temperature, and CO2 levels). OBJECTIVE: To compare proliferation and expression of osteoblastic phenotype of cells derived from vertebral lamina and iliac grafting. SUMMARY OF BACKGROUND DATA: Many factors play a role in the success of bone graft in spinal fusion including osteoblastic cell population. Two common sources of graft are vertebral lamina and iliac crest, however, differences in proliferation and osteoblastic phenotype expression between cells from these sites have not been investigated. METHODS: Cells obtained from cancellous bone of both vertebral lamina and iliac crest were cultured and proliferation was evaluated by direct cell counting and viability detected by Trypan blue. Alkaline phosphatase (ALP) activity was evaluated by thymolphthalein release from thymolphthalein monophosphate and matrix mineralization by staining with alizarin red S. Gene expression of ALP, osteocalcin, runt-related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, osteoprotegerin, and receptor activator of NF-kB ligand was analyzed by real-time PCR. All comparisons were donor-matched. RESULTS: Proliferation was greater at days 7 and 10 in cells from vertebral lamina compared with ones from iliac crest without difference in cell viability. ALP activity was higher in cells from vertebral lamina compared with cells from iliac crest at days 7 and 10. At 21 days, mineralized matrix was higher in cells derived from vertebral lamina than from iliac crest. At day 7, gene expression of ALP, osteocalcin, runt-related transcription factor 2, Msh homeobox 2, bone morphogenetic protein 7, intercellular adhesion molecule 1 precursor, receptor activator of NF-kB ligand, and osteoprotegerin was higher in cells derived from vertebral lamina compared with iliac crest. CONCLUSION: Cell proliferation and osteoblastic phenotype development in cells derived from cancellous bone were more exuberant in cultures of vertebral lamina than of iliac crest.

• Effect of growth hormone on in vitro osteogenesis and gene expression of human osteoblastic cells is donor-age-dependent.

  Authors: Grasiele E Crippa, Marcio M Beloti, Cristina R Cardoso, João S Silva, Adalberto L Rosa

  Journal of cellular biochemistry. 06/2008; 104(2):369-76.

  It has been demonstrated that the effect of GH on bone tissue is reduced with aging. In this study we tested the hypothesis that the action of GH on osteoblastic cells is donor-age-dependent byIt has been demonstrated that the effect of GH on bone tissue is reduced with aging. In this study we tested the hypothesis that the action of GH on osteoblastic cells is donor-age-dependent by investigating the effect of GH on the development of osteoblastic phenotype in cultures of cells from adolescents (13-16 years old), young adults (18-35 years old), and adults (36-49 years old). Osteoblastic cells derived from human alveolar bone were cultured with or without GH for periods of up to 21 days, and parameters of in vitro osteogenesis and gene expression of osteoblastic markers were evaluated. GH increased culture growth, collagen content and alkaline phosphatase (ALP) activity in cultures from adolescents and young adults, whereas non-significant effect was observed in cultures from adults. While GH significantly increased the bone-like formation in cultures from adolescents, a slightly effect was observed in cultures from young adults and no alteration was detected in cultures from adults. Results from real-time PCR demonstrated that GH upregulated ALP, osteocalcin, type I collagen, and Cbfa1 mRNA levels in cultures from adolescents. In addition, cultures from young adults showed higher ALP mRNA expression and the expression of all evaluated genes was not affected by GH in cultures from adults. These results indicate that the GH effect on both in vitro osteogenesis and gene expression of osteoblastic markers is donor-age-dependent, being more pronounced on cultures from adolescents.

• In vitro cytotoxicity of dental alloys and cpTi obtained by casting.

  Authors: Adriana C L Faria, Adalberto L Rosa, Renata C S Rodrigues, Ricardo F Ribeiro

  Journal of biomedical materials research. Part B, Applied biomaterials. 06/2008; 85(2):504-8.

  BACKGROUND: This study aimed to compare the cytotoxicity of base-metal dental alloys and to evaluate if the casting method could influence their cytotoxicity. METHODS: Disks of base-metal dentalBACKGROUND: This study aimed to compare the cytotoxicity of base-metal dental alloys and to evaluate if the casting method could influence their cytotoxicity. METHODS: Disks of base-metal dental alloys were cast by two methods: plasma, under argon atmosphere, injected by vacuum-pressure; and oxygen-gas flame, injected by centrifugation, except Ti-6Al-4V and commercially pure titanium (cpTi), cast only by plasma. SCC9 cells were cultured in culture media D-MEM/Ham's F12 supplemented, at 37 degrees C in a humidified atmosphere of 5% carbon dioxide and 95% air, on the previously prepared disks. At subconfluence in wells without disks (control), cell number and viability were evaluated. RESULTS: In plasma method, cpTi and Ti-6Al-4V were similar to control and presented higher number of cells than all other alloys, followed by Ni-Cr. In oxygen-gas flame method, all alloys presented fewer cells than control. Ni-Cr presented more cells than any other alloy, followed by Co-Cr-Mo-W which presented more cells than Ni-Cr-Ti, Co-Cr-Mo, and Ni-Cr-Be. There were no significant differences between casting methods related to cell number. Cell viability was not affected by either chemical composition or casting methods. CONCLUSION: cpTi and Ti-6Al-4V were not cytotoxic while Ni-Cr-Be was the most cytotoxic among tested alloys. The casting method did not affect cytotoxicity of the alloys.

• Chronic ethanol intake inhibits in vitro osteogenesis induced by osteoblasts differentiated from stem cells.

  Authors: Maria L Rosa, Marcio M Beloti, Natalia Prando, Regina H C Queiroz, Paulo T de Oliveira, Adalberto L Rosa

  Journal of applied toxicology : JAT. 04/2008; 28(2):205-11.

  The study investigated whether chronic ethanol (ETH) intake and subsequent ETH exposure of cell cultures affects osteoblast differentiation by evaluating key parameters of in vitro osteogenesis. RatsThe study investigated whether chronic ethanol (ETH) intake and subsequent ETH exposure of cell cultures affects osteoblast differentiation by evaluating key parameters of in vitro osteogenesis. Rats were treated with 5-20% (0.85-3.43 mm) ETH, increasing by 5% per week for a period of 4 weeks (habituation), after which the 20% level was maintained for 15 days (chronic intake). Bone-marrow stem cells from control (CONT) or ETH-treated rats were cultured in osteogenic medium which was either supplemented (ETH) or not supplemented (CONT) with 1.3 mm ethanol. Thus, four groups relating to rat treatment/culture supplementation were evaluated: (1) CONT/CONT, (2) ETH/CONT, (3) CONT/ETH and (4) ETH/ETH. Cell morphology, proliferation and viability, total protein content, alkaline phosphatase (ALP) activity and bone-like nodule formation were evaluated. Chronic ethanol intake significantly reduced both food and liquid consumption and body weight gain. No difference was seen in cell morphology among treatments. Cell number was affected at 7 and 10 days as follows: CONT/CONT = CONT/ETH < ETH/CONT = ETH/ETH. Doubling time between 3 and 10 days was greater in groups of CONT animals: ETH/ETH = ETH/CONT < CONT/ETH = CONT/CONT. Cell viability and ALP activity were not affected by either animal treatment or culture exposure to ethanol. At day 21, the total protein content was affected as follows: ETH/ETH = CONT/ETH < ETH/CONT = CONT/CONT. Bone-like nodule formation was affected as follows: ETH/ETH < CONT/ETH < ETH/CONT < CONT/CONT. These results show that chronic ethanol intake, followed by the exposure of osteoblasts to ethanol, inhibited the differentiation of osteoblasts, as indicated by an increased proliferation rate and reduced bone-like nodule formation.

• Microarray-based gene expression analysis of human osteoblasts in response to different biomaterials.

  Authors: Karina F Bombonato-Prado, Larissa S Bellesini, Cristina M Junta, Márcia M Marques, Geraldo A Passos, Adalberto L Rosa

  Journal of biomedical materials research. Part A. 03/2008;

  Several biomaterials have been widely used in bone regeneration/substitution procedures in orthopedic and oral surgery. However, how these biomaterials alter osteoblast gene expression is poorlySeveral biomaterials have been widely used in bone regeneration/substitution procedures in orthopedic and oral surgery. However, how these biomaterials alter osteoblast gene expression is poorly understood. We therefore attempted to address this question by using cDNA microarray technique to identify genes that are differentially regulated in osteoblasts exposed to biomaterials comprehending the biocompatibility spectrum of bioactive (bioglass and hydroxyapatite), bioinert (Ti and stainless steel), and biotolerant (polymethylmethacrylate). By using a cDNA microarray containing 687 human IMAGE sequences, we identified in primary cultures of osteoblastic cells differentiated from the human bone marrow and exposed to these biomaterials, genes whose expression was significantly upregulated or downregulated. Among the differentially expressed genes we have found those involved with cell cycle regulation, cell differentiation and proliferation, apoptosis, cell adhesion, bone mineralization and skeletal development. These results can be relevant to a better understanding of the molecular mechanism underlying the behavior of osteoblasts in bone regenerative procedures. (c) 2008 Wiley Periodicals, Inc. J Biomed Mater Res 2008.

• Culture of osteogenic cells from human alveolar bone: a useful source of alkaline phosphatase.

  Authors: Ana Maria S Simão, Marcio M Beloti, Adalberto L Rosa, Paulo T de Oliveira, José Mauro Granjeiro, João M Pizauro, Pietro Ciancaglini

  Cell biology international. 12/2007; 31(11):1405-13.

  The aim of this study was to obtain membrane-bound alkaline phosphatase from osteoblastic-like cells of human alveolar bone. Cells were obtained by enzymatic digestion and maintained in primaryThe aim of this study was to obtain membrane-bound alkaline phosphatase from osteoblastic-like cells of human alveolar bone. Cells were obtained by enzymatic digestion and maintained in primary culture in osteogenic medium until subconfluence. First passage cells were cultured in the same medium and at 7, 14, and 21 days, total protein content, collagen content, and alkaline phosphatase activity were evaluated. Bone-like nodule formation was evaluated at 21 days. Cells in primary culture at day 14 were washed with Tris-HCl buffer, and used to extract the membrane-bound alkaline phosphatase. Cells expressed osteoblastic phenotype. The apparent optimum pH for PNPP hydrolysis by the enzyme was pH 10.0. This enzyme also hydrolyzes ATP, ADP, fructose-1-phosphate, fructose-6-phosphate, pyrophosphate and beta-glycerophosphate. PNPPase activity was reduced by typical inhibitors of alkaline phosphatase. SDS-PAGE of membrane fraction showed a single band with activity of approximately 120 kDa that could be solubilized by phospholipase C or Polidocanol.

• Bone response to three different chemical compositions of fluorcanasite glass-ceramic.

  Authors: Valdemar Mallet da Rocha Barros, Jorge Luiz J Liporaci, Adalberto L Rosa, Marcela Caffarena Junqueira, Paulo Tambasco de Oliveira, Anthony Johnson, Richard van Noort

  Journal of biomedical materials research. Part A. 11/2007; 83(2):480-3.

  The aim of this study was to evaluate the bone response to three fluorcanasite glass-ceramic compositions with different solubilities (K3, K5, and K8) after implantation in a femur rabbit model.The aim of this study was to evaluate the bone response to three fluorcanasite glass-ceramic compositions with different solubilities (K3, K5, and K8) after implantation in a femur rabbit model. Fluorcanasite glass-ceramic rods were implanted bilaterally in the mid-shafts rabbit femurs. Implants were harvested at 8 and 12 weeks and prepared for histological and histomorphometric analyses at the light microscope level. Bioglass 45S5 rods were used as a control material. At 8 weeks, all fluorcanasite glass-ceramics were entirely surrounded by a nonmineralized connective tissue. At 12 weeks, reduced areas of bone tissue were observed in the cortical area in direct contact with the K3 and K5 fluorcanasite glass-ceramics compared to Bioglass 45S5, whereas no bone tissue was observed in direct contact with the K8 surface. Bone-to-implant contact in the cortical area was affected by the material chemical composition and ranked as follows: Bioglass 45S5>K3>K5>K8 (p=0.001). In the bone marrow, a layer of fibrous connective tissue formed in direct contact with the fluorcanasite glass-ceramics and Bioglass 45S5, and only rarely exhibited contact osteogenesis. All the fluorcanasite glass-ceramics appeared to degrade in the biological environment. The solubility ratio did not alter significantly the biological reply of the fluorcanasite glass-ceramics in vivo. Further modifications of the chemical composition of the fluorcanasite glass-ceramic are required to increase the stability of the material in vivo.

• In vitro biocompatibility of a novel membrane of the composite poly(vinylidene-trifluoroethylene)/barium titanate.

  Authors: Márcio M Beloti, Paulo T de Oliveira, Rossano Gimenes, Maria A Zaghete, Márcio J Bertolini, Adalberto L Rosa

  Journal of biomedical materials research. Part A. 12/2006; 79(2):282-8.

  This study was aimed at investigating the in vitro biocompatibility of a novel membrane of the composite poly(vinylidene-trifluoroethylene)/barium titanate (P(VDF-TrFE)/BT). Osteoblastic cells wereThis study was aimed at investigating the in vitro biocompatibility of a novel membrane of the composite poly(vinylidene-trifluoroethylene)/barium titanate (P(VDF-TrFE)/BT). Osteoblastic cells were obtained from human alveolar bone fragments and cultured under standard osteogenic condition until subconfluence. First passaged cells were cultured on P(VDF-TrFE)/BT and expanded polytetrafluoroethylene (e-PTFE--control) membranes in 24-well plates. Cell adhesion and spreading were evaluated at 30 min, and 4 and 24 h. For proliferation assay, cells were cultured for 1, 7, and 10 days. Cell viability was detected by trypan blue at 7 and 10 days. Total protein content and alkaline phosphatase (ALP) activity were measured at 7, 14, and 21 days. Cultures were stained with Alizarin red at 21 days, for detection of mineralized matrix. Data were compared by ANOVA and Student t test. Cell attachment (p = 0.001), cell number (p = 0.001), and ALP activity (p = 0.0001) were greater on P(VDF-TrFE)/BT. Additionally, doubling time was greater on P(VDF-TrFE)/BT (p = 0.03), indicating a decreased proliferation rate. Bone-like nodule formation took place only on P(VDF-TrFE)/BT. The present results showed that both membranes are biocompatible. However, P(VDF-TrFE)/BT presented a better in vitro biocompatibility and allowed bone-like nodule formation. Therefore, P(VDF-TrFE)/BT could be an alternative membrane to be used in guided tissue regeneration.

• Purmorphamine enhances osteogenic activity of human osteoblasts derived from bone marrow mesenchymal cells.

  Authors: Márcio M Beloti, Larissa S Bellesini, Adalberto L Rosa

  Cell biology international. 08/2005; 29(7):537-41.

  Purmorphamine is a novel small molecule with osteogenesis-inducing activity in multipotent mesenchymal progenitor cells, but there has been no evaluation of its effect on human cells to date. The aimPurmorphamine is a novel small molecule with osteogenesis-inducing activity in multipotent mesenchymal progenitor cells, but there has been no evaluation of its effect on human cells to date. The aim of this study was to investigate the induction of osteogenic activity by purmorphamine in human osteoblasts differentiated from bone marrow mesenchymal cells. Cells were cultured in 24-well plates at a density of 2x10(4)/well in medium containing 1, 2 or 3 microM purmorphamine, or vehicle. At 7, 14 and 21 days, cell proliferation, viability, and alkaline phosphatase (ALP) activity were evaluated. Bone-like nodule formation was evaluated at 21 days. Purmorphamine did not affect cell proliferation or viability, but increased ALP activity and bone-like nodule formation. These results indicate that events related to osteoblast differentiation, including increased ALP activity and bone-like nodule formation, are enhanced by purmorphamine.

• Osteoblastic differentiation of cultured rat bone marrow cells on hydroxyapatite with different surface topography.

  Authors: Adalberto L Rosa, Márcio M Beloti, Richard van Noort

  Dental materials : official publication of the Academy of Dental Materials. 12/2003; 19(8):768-72.

  Hydroxyapatite (HA) has been used in orthopedic, dental, and maxillofacial surgery as a bone substitute. OBJECTIVE: The aim of this investigation was to study the effect of surface topographyHydroxyapatite (HA) has been used in orthopedic, dental, and maxillofacial surgery as a bone substitute. OBJECTIVE: The aim of this investigation was to study the effect of surface topography produced by the presence of microporosity on the response of the rat bone marrow cells, evaluating: cell attachment, proliferation, total protein content, alkaline phosphatase (ALP) activity, and bone-like nodule formation. METHODS: Cells were cultured on HA discs manufactured by a combination of uniaxial powder pressing and different sintering conditions, with different percentage of microporosity (<5%-HA5, 15%-HA15, and 30%-HA30). For attachment evaluation, cells were cultured for 2 h. Proliferation was evaluated after 7 and 14 days. After 14 days, total protein content and ALP activity were measured. For bone-like nodule formation, cells were cultured for 21 days. Data were compared by ANOVA and Duncan's multiple range test when appropriate. RESULTS: Cell attachment was not affected by surface topography (p=0.37). Proliferation (p=0.001), total protein content (p=0.039), ALP activity (p=0.050), and bone-like nodule formation (p=0.00001) were all significantly decreased by the most irregular surface (HA30). SIGNIFICANCE. These results suggest that initial cell events were not affected by the surface topography of the HA. However, intermediary and final events such as proliferation, protein synthesis, ALP activity, and bone-like nodule formation favored surfaces with a more regular topography, such as that presents in HA with 15% or less of microporosity.

• In vivo biocompatibility of three different chemical compositions of Ricinus communis polyurethane.

  Authors: Valdemar M R Barros, Adalberto L Rosa, Márcio M Beloti, Gilberto Chierice

  Journal of biomedical materials research. Part A. 11/2003; 67(1):235-9.

  Alteration in the chemical composition of a biomaterial may be undertaken to improve its biological properties. The aim of this work was to evaluate the biocompatibility of three chemicalAlteration in the chemical composition of a biomaterial may be undertaken to improve its biological properties. The aim of this work was to evaluate the biocompatibility of three chemical compositions of Ricinus communis polyurethane (RCP): RCPp (pure RCP), RCP + CaCO(3), and RCP +Ca(3)(PO(4))(2). RCP cylinders were surgically implanted in rabbit femurs. After 8, 12, and 16 weeks, the femurs were removed, fixed, sectioned, ground, and stained by Stevenel's blue/Alizarin red S for light microscopy and histomorphometry. The osseointegration and osseoconductivity were calculated by means of image analysis and the data were submitted to the Kruskal-Wallis test followed by Dunn's test. Osseointegration was already completed after 8 weeks on RCP + Ca(3)(PO(4))(2) because similar values were found from week 8 to 16, whereas it showed a time-dependent increase on RCPp and RCP +CaCO(3). The osseointegration was greater on RCP + Ca(3)(PO(4))(2) in all periods when compared with RCPp, and after 8 and 12 weeks when compared with RCP + CaCO(3). None of the RCP samples presented osseoconductivity. The present results showed that RCP blended with calcium phosphate improved the biocompatibility by both enhancing and accelerating its osseointegration. Based on the absence of osseoconductivity, RCP was considered to be a bioinert material.

• TAK-778 enhances osteoblast differentiation of human bone marrow cells.

  Authors: Adalberto L Rosa, Marcio M Beloti

  Journal of cellular biochemistry. 09/2003; 89(6):1148-53.

  TAK-778 has been shown to induce bone growth in in vitro and in vivo models. However, there are no studies evaluating the effect of TAK-778 on human cells. Thus, the aim of this study was toTAK-778 has been shown to induce bone growth in in vitro and in vivo models. However, there are no studies evaluating the effect of TAK-778 on human cells. Thus, the aim of this study was to investigate osteogenesis induced by TAK-778 on human bone marrow cells. Cells were cultured in 24-well culture plates at a cell density of 2 x 10(4) cells/well in culture medium containing TAK-778 (10(-7), 10(-6), and 10(-5) M, each) or vehicle. During the culture period, cells were incubated at 37 degrees C in a humidified atmosphere of 5% CO(2) and 95% air. For attachment evaluation, cells were cultured for 4 and 24 h. After 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity, and bone-like formation were evaluated. Data were compared by ANOVA and Duncan's multiple range test. TAK-778 did not affect cell attachment and viability. Cell number was reduced by TAK-778 in all time period evaluated in a dose-dependent way. The effect of TAK-778 on total protein content, ALP activity and bone-like formation was a dose-dependent increase. The present results suggest that initial cell events such as cell attachment are not affected by TAK-778 while events that indicate osteoblast differentiation including reduced cell proliferation, and increased both ALP activity and bone-like formation are enhanced by TAK-778 in a time and dose-dependent way. It means that TAK-778 could be a useful drug to enhance new bone formation in clinical situations that require rapid restoration of physiologic function, such as orthopedic and maxillofacial surgery.

• TAK-778 enhances osteoblast differentiation of human bone marrow cells cultured on titanium.

  Authors: Adalberto L Rosa, Márcio M Beloti

  Biomaterials. 09/2003; 24(17):2927-32.

  TAK-778 induces bone growth in in vitro and in vivo models. The aim of this study was to evaluate the osteogenic potential of TAK-778 on human bone marrow cells cultured on commercially pure titaniumTAK-778 induces bone growth in in vitro and in vivo models. The aim of this study was to evaluate the osteogenic potential of TAK-778 on human bone marrow cells cultured on commercially pure titanium (cpTi). Cells were cultured either in absence or in presence of TAK-778 (10(-5)M) on cpTi in supplemented alpha-MEM. For attachment evaluation, cells were cultured for 4 and 24h. After 7, 14, and 21 days, cell proliferation, cell viability, total protein content, alkaline phosphatase (ALP) activity, and bone-like formation were evaluated. TAK-778 did not affect cell attachment and viability. Cell number was reduced by TAK-778. ALP activity, total protein content, and bone-like formation were increased by TAK-778. These results suggest that initial cell events such as cell attachment are not affected by TAK-778 while events that indicate osteoblast differentiation including reduced cell proliferation, and increased both ALP activity and bone-like formation are enhanced by TAK-778 in presence of cpTi. It means that TAK-778 could be a useful drug to improve the osseointegration of implants by both enhancing and accelerating bone formation on Ti surface.

• Response of rat bone marrow cells to commercially pure titanium submitted to different surface treatments.

  Authors: Samuel P Xavier, Paulo S P Carvalho, Márcio M Beloti, Adalberto L Rosa

  Journal of dentistry. 04/2003; 31(3):173-80.

  OBJECTIVES: Alterations in the commercially pure titanium (cpTi) surface may be undertaken to improve its biological properties. The aim of this study is to investigate the biocompatibility of cpTiOBJECTIVES: Alterations in the commercially pure titanium (cpTi) surface may be undertaken to improve its biological properties. The aim of this study is to investigate the biocompatibility of cpTi submitted to different surface treatments. METHODS: The cpTi surfaces were prepared so that machined and blasted surfaces, either acid etched or not, were compared using rat bone marrow cells cultured to differentiated into osteoblast. For attachment evaluation, cells were cultured for 4 and 24h. Cell morphology was evaluated after 3 days. After 7, 14, and 21 days cell proliferation was evaluated. Total protein content and alkaline phosphatase (ALP) activity were evaluated after 14 and 21 days. For bone-like nodule formation, cells were cultured for 21 days. Data were compared by analysis of variance. RESULTS: Cell attachment, cell morphology, cell proliferation, and ALP activity were not affected by surface treatments. Total protein content was reduced by blasted and acid etched surface. Bone-like nodule formation was significantly reduced by blasted, acid etched, and a combination of both blasted and acid etched surfaces. CONCLUSIONS: Based on these results, it can be suggested that cpTi surfaces that were submitted only to machining treatment favor the final event of osteoblastic differentiation of the rat bone marrow cells, evidenced by increased bone-like nodule formation.

• Rat bone marrow cell response to titanium and titanium alloy with different surface roughness.

  Authors: Adalberto L Rosa, Márcio M Beloti

  Clinical oral implants research. 03/2003; 14(1):43-8.

  In general, cell response is affected by both chemical composition and surface roughness of implant materials. The aim of this study was to evaluate the effect of titanium (Ti) chemical compositionIn general, cell response is affected by both chemical composition and surface roughness of implant materials. The aim of this study was to evaluate the effect of titanium (Ti) chemical composition and surface roughness on the response of rat bone marrow cells, examining cell attachment, cell proliferation, total protein content, alkaline phosphatase (ALP) activity, and bone-like nodule formation. Cells were cultured on both commercially pure titanium (cpTi) and titanium-6-aluminum-4-vanadium alloy (Ti-A) discs with four different average roughnesses (Ra). For attachment evaluation, cells were cultured for 2 h. After 14 days, cell proliferation, total protein content, and ALP activity were evaluated. Bone-like nodule formation was evaluated after 21 days. Data were compared by anova and Duncan's multiple range test when appropriate. Cell attachment and total protein content were affected by neither Ti chemical composition (P = 0.201, and P = 0.639, respectively) or surface roughness (P = 0.972, and P = 0.660, respectively). Proliferation, ALP activity, and bone-like nodule formation were affected only by Ti chemical composition (P = 0.0001, P = 0.064, and P = 0.0001, respectively). These results suggest that cpTi would optimize osteoblastic differentiation by rat bone marrow cells, including reduced cell proliferation, and increased ALP activity and bone-like nodule formation, while surface roughness, within the Ra parameters used, would not affect significantly the rat bone marrow cell response.

• Effect of the chemical composition of Ricinus communis polyurethane on rat bone marrow cell attachment, proliferation, and differentiation.

  Authors: Márcio M Beloti, Karen R N Hiraki, Valdemar M R Barros, Adalberto L Rosa

  Journal of biomedical materials research. Part A. 02/2003; 64(1):171-6.

  Alterations in the chemical composition of a polymer may be undertaken to improve its biological properties. The aim of this study was to investigate the in vitro biocompatibility of Ricinus communisAlterations in the chemical composition of a polymer may be undertaken to improve its biological properties. The aim of this study was to investigate the in vitro biocompatibility of Ricinus communis polyurethane (RCP) with three different chemical compositions: RCPp (pure RCP), RCP+CaCO(3), and RCP+Ca(3)(PO(4))(2). Rat bone marrow cells were cultivated under conditions that allowed osteoblastic differentiation and were evaluated for cell attachment, cell proliferation, cell morphology, total protein content, alkaline phosphatase (ALP) activity, and bonelike nodule formation. For the evaluation of attachment, cells were cultured for 4 h. After 3 days, cell morphology was evaluated. Cell proliferation was evaluated after 7 and 14 days. Total protein content and ALP activity were evaluated after 14 days. For bonelike nodule formation, cells were cultured for 21 days. Data were compared with an analysis of variance and Duncan's multiple range test when appropriate. Cell attachment and ALP activity were not affected by RCP chemical composition. Proliferation, total protein content, and bonelike nodule formation were all affected by RCP chemical composition. These results suggest that initial cell events are not affected by RCP chemical composition, whereas RCPs blended with calcium carbonate or, better yet, calcium phosphate, by favoring events that promote matrix mineralization, are more biocompatible materials.

• Response of rat bone marrow cells to commercially pure titanium submitted to different surface treatments

  Authors: Samuel P. Xavier, Paulo S.P. Carvalho, Márcio M. Beloti, Adalberto L. Rosa

  Journal of Dentistry.

  Objectives. Alterations in the commercially pure titanium (cpTi) surface may be undertaken to improve its biological properties. The aim of this study is to investigate the biocompatibility of cpTiObjectives. Alterations in the commercially pure titanium (cpTi) surface may be undertaken to improve its biological properties. The aim of this study is to investigate the biocompatibility of cpTi submitted to different surface treatments.Methods. The cpTi surfaces were prepared so that machined and blasted surfaces, either acid etched or not, were compared using rat bone marrow cells cultured to differentiated into osteoblast. For attachment evaluation, cells were cultured for 4 and 24 h. Cell morphology was evaluated after 3 days. After 7, 14, and 21 days cell proliferation was evaluated. Total protein content and alkaline phosphatase (ALP) activity were evaluated after 14 and 21 days. For bone-like nodule formation, cells were cultured for 21 days. Data were compared by analysis of variance.Results. Cell attachment, cell morphology, cell proliferation, and ALP activity were not affected by surface treatments. Total protein content was reduced by blasted and acid etched surface. Bone-like nodule formation was significantly reduced by blasted, acid etched, and a combination of both blasted and acid etched surfaces.Conclusions. Based on these results, it can be suggested that cpTi surfaces that were submitted only to machining treatment favor the final event of osteoblastic differentiation of the rat bone marrow cells, evidenced by increased bone-like nodule formation.

• Nitric acid passivation does not affect in vitro biocompatibility of titanium.

  Authors: Adriana C L Faria, Márcio M Beloti, Adalberto L Rosa

  The International journal of oral & maxillofacial implants. 18(6):820-5.

  PURPOSE: In general, both chemical composition and surface features of implants affect cell response. The aim of this study was to evaluate the effect of titanium (Ti) passivation on the response ofPURPOSE: In general, both chemical composition and surface features of implants affect cell response. The aim of this study was to evaluate the effect of titanium (Ti) passivation on the response of rat bone marrow cells, considering cell attachment, cell morphology, cell proliferation, total protein content, alkaline phosphatase (ALP) activity, and bonelike nodule formation. MATERIALS AND METHODS: Cells were cultured on both commercially pure titanium (cpTi) and titanium-aluminium-vanadium alloy (Ti-6Al-4V) discs, either passivated or not. For attachment evaluation, cells were cultured for 4 and 24 hours. Cell morphology was evaluated after 4 days. After 7, 14, and 21 days, cell proliferation, total protein content, and ALP activity were evaluated. Bonelike nodule formation was evaluated after 21 days. Data were compared by analysis of variance and the Duncan multiple range test. RESULTS: Cell attachment, cell morphology, cell proliferation, total protein content, ALP activity, and bonelike nodule formation all were unaffected by Ti composition or passivation. DISCUSSION AND CONCLUSION: Although the protocol for passivation used here could interfere with the pattern of ions released from Ti-6Al-4V and cpTi surfaces, the present study did not show any effect of this surface treatment on in vitro biocompatibility of Ti as evaluated by osteoblast attachment, proliferation, and differentiation.