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Histological and Nanomechanical Properties of a New Nanometric Hydroxiapatite Implant Surface. An In Vivo Study in Diabetic Rats

Oliveira, Paula G F P; Coelho, Paulo G; Bergamo, Edmara T P; Witek, Lukasz; Borges, Cristine A; Bezerra, Fábio B; Novaes, Arthur B; Souza, Sergio L S
Implant therapy is a predictable treatment to replace missing teeth. However, the osseointegration process may be negatively influenced by systemic conditions, such as diabetes mellitus (DM). Microtopography and implant surface developments are strategies associated to better bone repair. This study aimed to evaluate, in healthy and diabetic rats, histomorphometric (bone to implant contact = %BIC; and bone area fraction occupancy = %BAFO) and nanomechanical (elastic modulus = EM; and hardness = H) bone parameters, in response to a nanometric hydroxyapatite implant surface. Mini implants (machined = MAC; double acid etched = DAE, and with addition of nano-hydroxyapatite = NANO) were installed in tibias of healthy and diabetic rats. The animals were euthanized at 7 and 30 days. NANO surface presented higher %BIC and %BAFO when compared to MAC and DAE (data evaluated as a function of implant surface). NANO surface presented higher %BIC and %BAFO, with statistically significant differences (data as a function of time and implant surface). NANO surface depicted higher EM and H values, when compared to machined and DAE surfaces (data as a function of time and implant surface). Nano-hydroxyapatite coated implants presented promising biomechanical results and could be an important tool to compensate impaired bone healing reported in diabetics.
PMID: 33322243
ISSN: 1996-1944
CID: 4726652

Microstructural, mechanical, and optical characterization of an experimental aging-resistant zirconia-toughened alumina (ZTA) composite

Lopes, A C O; Coelho, P G; Witek, L; Benalcázar Jalkh, E B; Gênova, L A; Monteiro, K N; Cesar, P F; Lisboa-Filho, P N; Bergamo, E T P; Ramalho, I S; Campos, T M B; Bonfante, E A
OBJECTIVE:(ZTA Zpex) compared to a translucent zirconia (Zpex) and Alumina. METHODS:Disc-shaped specimens were obtained by uniaxial and isostatic pressing the synthesized powders (n = 70/material). After sintering and polishing, half of the specimens underwent aging (20 h, 134 °C, 2.2 bar). Crystalline content and microstructure were evaluated using X-ray diffraction and scanning electron microscopy, respectively. Specimens underwent biaxial flexural strength testing to determine the characteristic stress, Weibull modulus, and reliability. Translucency parameter (TP) and Contrast ratio (CR) were calculated to characterize optical properties. RESULTS:ZTA Zpex demonstrated a compact surface with a uniform dispersion of zirconia particles within the alumina matrix, and typical alumina and zirconia crystalline content. ZTA Zpex and alumina exhibited higher CR and lower TP than Zpex. ZTA Zpex and Zpex showed significantly higher characteristic stress relative to alumina. While aging did not affect optical and mechanical properties of ZTA Zpex and alumina, Zpex demonstrated a significant increase in translucency, as well as a in characteristic stress. Alumina reliability was significantly lower than others at 300 MPa, ZTA Zpex and Zpex reliability decreased at 800 MPa, except for aged Zpex. SIGNIFICANCE/CONCLUSIONS:While aging did not affect the mechanical nor the optical properties of ZTA Zpex and alumina, it did alter both properties of Zpex. The results encourage further investigations to engineer ZTA as a framework material for long span fixed dental prostheses specially where darkened substrates, such as titanium implant abutments or endodontically treated teeth, demand masking.
PMID: 32943230
ISSN: 1879-0097
CID: 4629852

Three-Dimensional Printing for Craniofacial Bone Tissue Engineering

Shen, Chen; Witek, Lukasz; Flores, Roberto L; Tovar, Nick; Torroni, Andrea; Coelho, Paulo G; Kasper, F Kurtis; Wong, Mark; Young, Simon
The basic concepts from the fields of biology and engineering are integrated into tissue engineering to develop constructs for the repair of damaged and/or absent tissues, respectively. The field has grown substantially over the past two decades, with particular interest in bone tissue engineering (BTE). Clinically, there are circumstances in which the quantity of bone that is necessary to restore form and function either exceeds the patient's healing capacity or bone's intrinsic regenerative capabilities. Vascularized osseous or osteocutaneous free flaps are the standard of care with autologous bone remaining the gold standard, but is commonly associated with donor site morbidity, graft resorption, increased operating time, and cost. Regardless of the size of a craniofacial defect, from trauma, pathology, and osteonecrosis, surgeons and engineers involved with reconstruction need to consider the complex three-dimensional (3D) geometry of the defect and its relationship to local structures. Three-dimensional printing has garnered significant attention and presents opportunities to use craniofacial BTE as a technology that offers a personalized approach to bony reconstruction. Clinicians and engineers are able to work together to produce patient-specific space-maintaining scaffolds tailored to site-specific defects, which are osteogenic, osseoconductive, osseoinductive, encourage angiogenesis/vasculogenesis, and mechanically stable upon implantation to prevent immediate failure. In this work, we review biological and engineering principles important in applying 3D printing technology to BTE for craniofacial reconstruction as well as present recent translational advancements in 3D printed bioactive ceramic scaffold technology.
PMCID:7759279
PMID: 32842918
ISSN: 1937-335x
CID: 4751522

Influence of Abutment Fabrication Method on 3D Fit at the Implant-Abutment Connection

Ramalho, Ilana; Witek, Lukasz; Coelho, Paulo G; Bergamo, Edmara; Pegoraro, Luiz Fernando; Bonfante, Estevam A
PURPOSE/OBJECTIVE:To three-dimensionally evaluate the internal fit at the implant-abutment interface of abutments fabricated with different workflows using a combination of the silicone replica technique and microcomputed tomography (μCT). MATERIALS AND METHODS/METHODS:) were assessed. Data were statistically evaluated using ANOVA and Tukey test (P < .05). RESULTS:in the full digital group, which exhibited higher mean values compared to the other regions (P = .000). The 3D measurements for quantification of internal discrepancy were strongly associated with the 2D measurements. CONCLUSION/CONCLUSIONS:Ti-Base and UCLA abutments exhibited better internal fit at the implant-abutment interfaces compared to a fully digitalized workflow (CAD/CAM custom abutments).
PMID: 33284906
ISSN: 1942-4426
CID: 4736802

WE43 and WE43-T5 Mg alloys screws tested in-vitro cellular adhesion and differentiation assay and in-vivo histomorphologic analysis in an ovine model

Torroni, Andrea; Witek, Lukasz; Fahliogullari, Hayat Pelin; Bortoli, Joao Paulo; Ibrahim, Amel; Hacquebord, Jacques; Gupta, Nikhil; Coelho, Paulo
WE43 Mg alloy proved to be an ideal candidate for production of resorbable implants in both clinical and trial settings. In previous studies we tested biocompatibility and degradation properties of WE43 (as-cast) and artificially aged (WE43-T5) Mg alloys in a sheep model. Both alloys showed excellent biocompatibility with the as-cast, WE43, form showing increased degradability compared to the artificially aged, WE43-T5. In the present study, our group assessed the biological behavior and degradation pattern of the same alloys when implanted as endosteal implants in a sheep model. Twelve screws (3x15 mm) were evaluated, one screw per each composition was placed bi-cortically in the mandible of each animal with a titanium (2x12 mm) screw serving as an internal positive control. At 6 and 24 weeks histomorphological analysis was performed, at 6 weeks as cast, WE43, yielded a higher degradation rate, increased bone remodeling and osteolysis compared to the WE43-T5 alloy; however, at 24 weeks WE43-T5 showed higher degradation rate and increased bone remodeling than as-cast. In vitro assay of cell growth, adhesion and differentiation was also conducted to investigate possible mechanisms underlying the behavior expressed from the alloys in vivo. In conclusion WE43-T5 indicated bone/implant interaction properties that makes it more suitable for fabrication of endosteal bone screws.
PMID: 32903065
ISSN: 1530-8022
CID: 4629792

Aging resistant ZTA composite for dental applications: Microstructural, optical and mechanical characterization

Benalcazar Jalkh, Ernesto Byron; Monteiro, Kelli Nunes; Cesar, Paulo Francisco; Genova, Luis Antonio; Bergamo, Edmara T P; Lopes, Adolfo Coelho de Oliveira; Lima, Erick; Lisboa-Filho, Paulo Noronha; Campos, Tiago Moreira Bastos; Witek, Lukasz; Coelho, Paulo G; Borges, Ana Flavia Sanches; Bonfante, Estevam A
OBJECTIVE:To synthesize a zirconia toughened alumina (ZTA) composite with 70% alumina reinforced by 30% zirconia for dental applications and to characterize its microstructure and optical properties for comparison with the isolated counterpart materials and a first-generation 3Y-TZP. METHODS:Disc-shaped specimens were divided in four groups (n = 70/material): (1) 3YSB-E (first generation 3Y-TZP), (2) Zpex (second generation 3Y-TZP), (3) alumina, and (4) ZTA-Zpex 70/30. After synthesis, ceramic powders were pressed, and green-body samples sintered following a predetermined protocol. Specimens were polished to obtain a mirror surface finish. Apparent density was measured by Archimedes principle. X-ray diffraction (XRD) and scanning electron microscope (SEM) were used to characterize the crystalline content and microstructure. Reflectance tests were performed to determine the contrast-ratio (CR) and translucency-parameter (TP). Mechanical properties were assessed by biaxial flexural strength (BFS) test. All analyses were conducted before and after artificial aging (20 h, 134 °C, 2.2 bar). Optical parameters were evaluated through repeated-measures analysis of variance and Tukey tests (p < 0.05). BFS data were analyzed using Weibull statistics (95% CI). RESULTS:High density values (95-99%) were found for all ceramic materials and SEM images exhibited a dense microstructure. While XRD patterns revealed the preservation of crystalline content in the ZTA composite, an increase in the monoclinic peak was observed for pure zirconias after aging. Significantly higher CR and lower TP values were observed for the ZTA composite, followed by alumina, 3YSB-E, and Zpex. The highest characteristic stress was recorded for 3YSB-E, followed by intermediate values between ZTA and Zpex, and the lowest for alumina. Aging affected the optical and mechanical properties of both zirconias, while remained stable for ZTA composite and alumina. SIGNIFICANCE/CONCLUSIONS:The synthesis of experimental 70-30% ZTA composite was successful and its relevance for dental applications relies on its higher masking ability, aging resistance, and strength similar to zirconia.
PMID: 32561117
ISSN: 1879-0097
CID: 4510792

Obesity/Metabolic Syndrome and Diabetes Mellitus on Peri-implantitis

de Oliveira, Paula Gabriela Faciola Pessôa; Bonfante, Estevam A; Bergamo, Edmara T P; de Souza, Sérgio Luis Scombatti; Riella, Leonardo; Torroni, Andrea; Benalcazar Jalkh, Ernesto B; Witek, Lukasz; Lopez, Christopher D; Zambuzzi, Willian Fernando; Coelho, Paulo G
Literature has reported that up to 50% of dental implants may be affected by peri-implantitis, a bacteria-induced chronic inflammatory process, which promotes osteoclast-mediated bone resorption and inhibits bone formation, leading to progressive bone loss around implants. Current evidence points toward an increased risk for the development of peri-implantitis in both obesity/metabolic syndrome (MetS) and diabetes mellitus (DM) conditions relative to the healthy population. Currently, there is no effective treatment for peri-implantitis and the 50% prevalence in MetS and DM, along with its predicted increase in the worldwide population, presents a major concern in implant dentistry as hyperglycemic conditions are associated with bone-healing impairment; this may be through dysfunction of osteocalcin-induced glucose metabolism. The MetS/DM proinflammatory systemic condition and altered immune/microbiome response affect both catabolic and anabolic events of bone-healing that include increased osteoclastogenesis and compromised osteoblast activity, which could be explained by the dysfunction of insulin receptor that led to activation of signals related with osteoblast differentiation. Furthermore, chronic hyperglycemia along with associated micro- and macro-vascular ailments leads to delayed/impaired wound healing due to activation of pathways that are particularly important in initiating events linked to inflammation, oxidative stress, and cell apoptosis; this may be through deactivation of AKT/PKB protein, which possesses a pivotal role in drive survival and eNOS signaling. This review presents an overview of the local and systemic mechanisms synergistically affecting bone-healing impairment in MetS/DM individuals, as well as a rationale for hierarchical animal model selection, in an effort to characterize peri-implantitis disease and treatment.
PMID: 32591106
ISSN: 1879-3061
CID: 4510852

Clinical, histological, and nanomechanical parameters of implants placed in healthy and metabolically compromised patients

Granato, Rodrigo; Bergamo, Edmara T P; Witek, Lukasz; Bonfante, Estevam A; Marin, Charles; Greenberg, Michael; Kurgansky, Gregory; Coelho, Paulo G
OBJECTIVES/OBJECTIVE:To evaluate the clinical outcomes, histological parameters, and bone nanomechanical properties around implants retrieved from healthy and metabolic syndrome (MS) patients. METHODS:Twenty-four patients with edentulous mandibles (12/condition), received four implants between the mental foramina. An additional implant prototype was placed for retrieval histology. The following clinical outcomes were evaluated: insertion torque (IT), implant stability quotient (ISQ) values at baseline and after 60 days of healing, and implant survival. The prototype was retrieved after the healing and histologically processed for bone morphometric evaluation of bone-to-implant contact (%BIC) and bone area fraction occupancy (%BAFO), and bone nanoindentation to determine the elastic modulus (Em) and hardness (H). Descriptive statistical procedures and survival tests were used to analyze the data. RESULTS:The final study population was comprised of 10 women and 11 men (∼64 years). A total of 105 implants were placed, 21 retrieved for histology. Implant survival rates were similar between groups (>99 %). Similarly, IT and ISQ analyses showed no significant association with systemic condition (p > 0.216). Histological micrographs depicted similar bone morphology, woven bone, for both conditions. While MS (33 ± 5.3 %) and healthy (39 ± 6.5 %) individuals showed no significant difference for %BIC (p = 0.116), significantly higher %BAFO was observed for healthy (45 ± 4.6 %) relative to MS (30 ± 3.8 %) (p < 0.001). No significant differences on bone nanomechanical properties was observed (p > 0.804). CONCLUSIONS:Although no significant influence on clinical parameters and bone nanomechanical properties was observed, MS significantly reduced bone formation in the peri-implant area in the short-term. CLINICAL SIGNIFICANCE/CONCLUSIONS:A lower amount of bone formation in the peri-implant area was observed in comparison to healthy patients, although the other short-term clinical outcomes were not significantly different. Considering the escalating prevalence of MS patients in need for implant treatment, it becomes crucial to understand bone-to-implant response to determine the ideal loading time in this population.
PMID: 32738285
ISSN: 1879-176x
CID: 4572202

3D Printing of Microgel-Loaded Modular Microcages as Instructive Scaffolds for Tissue Engineering

Subbiah, Ramesh; Hipfinger, Christina; Tahayeri, Anthony; Athirasala, Avathamsa; Horsophonphong, Sivaporn; Thrivikraman, Greeshma; França, Cristiane Miranda; Cunha, Diana Araujo; Mansoorifar, Amin; Zahariev, Albena; Jones, James M; Coelho, Paulo G; Witek, Lukasz; Xie, Hua; Guldberg, Robert E; Bertassoni, Luiz E
Biomaterial scaffolds have served as the foundation of tissue engineering and regenerative medicine. However, scaffold systems are often difficult to scale in size or shape in order to fit defect-specific dimensions, and thus provide only limited spatiotemporal control of therapeutic delivery and host tissue responses. Here, a lithography-based 3D printing strategy is used to fabricate a novel miniaturized modular microcage scaffold system, which can be assembled and scaled manually with ease. Scalability is based on an intuitive concept of stacking modules, like conventional toy interlocking plastic blocks, allowing for literally thousands of potential geometric configurations, and without the need for specialized equipment. Moreover, the modular hollow-microcage design allows each unit to be loaded with biologic cargo of different compositions, thus enabling controllable and easy patterning of therapeutics within the material in 3D. In summary, the concept of miniaturized microcage designs with such straight-forward assembly and scalability, as well as controllable loading properties, is a flexible platform that can be extended to a wide range of materials for improved biological performance.
PMID: 32700332
ISSN: 1521-4095
CID: 4542662

Assessing osseointegration of metallic implants with boronized surface treatment

Witek, L; Tovar, N; Lopez, C-D; Morcos, J; Bowers, M; Petrova, R-S; Coelho, P-G
BACKGROUND:Modification of endosteal implants through surface treatments have been investigated to improve osseointegration. Boronization has demonstrated favorable mechanical properties, but limited studies have assessed translational, in vivo outcomes. This study investigated the effect of implant surface boronization on bone healing. MATERIAL AND METHODS/METHODS:Two implant surface roughness profiles (acid etched, machined) in CP titanium (type II) alloy implants were boronized by solid-state diffusion until 10-15µm boron coating was achieved. The surface-treated implants were placed bilaterally into 5 adult sheep ilia for three and six weeks. Four implant groups were tested: boronized machined (BM), boronized acid-etched (BAA), control machined (CM), and control acid-etched (CAA). Osseointegration was quantified by calculating bone to implant contact (BIC) and bone area fraction occupancy (BAFO). RESULTS:Both implant types treated with boronization had BIC values not statistically different from machined control implants at t=3 weeks, and significantly less than acid-etched control (p<0.02). BAFO values were not statistically different for all 3-week groups except machined control (significantly less at p <0.02). BAFO had a significant downward trend from 3 to 6 weeks in both boronized implant types (p<0.03) while both control implant types had significant increases in BIC and BAFO from 3 to 6 weeks. CONCLUSIONS:Non-decalcified histology depicted intramembranous-like healing/remodeling in bone for controls, but an absence of this dynamic process in bone for boronized implants. These findings are inconsistent with in vitro work describing bone regenerative properties of elemental Boron and suggests that effects of boron on in vivo bone healing warrant further investigation.
PMID: 32271322
ISSN: 1698-6946
CID: 4379132