Alejandro Pelaez-Vargas

Isotropic micropatterned silica coatings on zirconia induce guided cell growth for dental implants

UNLABELLED Titanium implants are the gold standard in dentistry; however, problems such as gingival tarnishing and peri-implantitis have been reported. For zirconia to become a competitive alternative dental implant material, surface modification techniques that induce guided tissue growth must be developed. OBJECTIVES To develop alternative surface modification techniques to promote guided tissue regeneration on zirconia materials, for applications in dental implantology. METHODS A methodology that combined soft lithography and sol-gel chemistry was used to obtain isotropic micropatterned silica coatings on yttria-stabilized zirconia substrates. The materials were characterized via chemical, structural, surface morphology approaches. In vitro biological behavior was evaluated in terms of early adhesion and viability/metabolic activity of human osteoblast-like cells. Statistical analysis was conducted using one-way ANOVA/Tukey HSD post hoc test. RESULTS Isotropic micropatterned silica coatings on yttria-stabilized zirconia substrates were obtained using a combined approach based on sol-gel technology and soft lithography. Micropatterned silica surfaces exhibited a biocompatible behavior, and modulated cell responses (i.e. inducing early alignment of osteoblast-like cells). After 7d of culture, the cells fully covered the top surfaces of pillar microstructured silica films. SIGNIFICANCE The micropatterned silica films on zirconia showed a biocompatible response, and were capable of inducing guided osteoblastic cell adhesion, spreading and propagation. The results herein presented suggest that surface-modified ceramic implants via soft lithography and sol-gel chemistry could potentially be used to guide periodontal tissue regeneration, thus promoting tight tissue apposition, and avoiding gingival retraction and peri-implantitis.

Micropatterned silica thin films with nanohydroxyapatite micro-aggregates for guided tissue regeneration

UNLABELLED Surface modification of biomaterials has been shown to improve the biological response to dental implants. The ability to create a controlled micro-texture on the implant via additive surface modification techniques with bioactive nanohydroxyapatite (nanoHA) may positively influence guided tissue regeneration. OBJECTIVE The main goal of this study was to produce micro-fabricated SiO(2) surfaces modified with nanohydroxyapatite particles and to characterize their influence on the biological response of Human Dental-Pulp Mesenchymal Stem Cells (hDP-MSCs) and Streptococcus mutans. MATERIALS AND METHODS A combined methodology of sol-gel and soft-lithography was used to produce micropatterned SiO(2) thin films with different percentages of nanoHA micro-aggregates. The surfaces were characterized by SEM/EDS, FT-IR/ATR, AFM, XPS quantitative elemental percentage and contact angle measurements. Biological characterization was performed using hDP-MSCs cultures, while Streptococcus mutans was the selected microorganism to evaluate the bacterial adhesion on the thin films. RESULTS Micropatterned SiO(2) surfaces with 0%, 1% and 5% of nanoHA micro-aggregates were successfully produced using a combination of sol-gel and soft-lithography. These surfaces controlled the biological response, triggering alignment and oriented proliferation of hDP-MSCs and significant differences in the adhesion of S. mutans to the different surfaces. SIGNIFICANCE The micropatterned surfaces exhibited biocompatible behavior that induced an oriented adhesion and proliferation of hDP-MSCs while SiO(2) presented low bacterial adhesion. These results show that the combination of sol-gel with soft-lithography is a good approach to create micropatterned surfaces with bioactive nanoparticles for guided tissue regeneration.

Palate morphology of bruxist children with mixed dentition. A pilot study

The objective of the study was to analyse quantitatively palatal morphology in bruxist and non-bruxist children with mixed dentition. Twenty-three children with mixed dentition were classified as bruxist according to their anxiety level, audible occlusal sounds related by the parents and signs of temporomandibular disorders; 23 children were control subjects matched for gender, age, and dental formula. The maxillary dental arches of all subjects were reproduced from alginate impressions cast in dental stone with a standardized technique. The casts were digitalized and mathematical equations were used to obtain the form of the palate in the sagittal, frontal and horizontal planes. Bruxist children had a statistically significant longer palate in the sagittal plane than control children; palatal shape differed especially in correspondence of the third, fourth and fifth teeth, bruxist children showing a relatively higher palate than control children. In this pilot study, sagittal plane differences in the palate between bruxist and non-bruxist children matched for age and gender were found. Further investigations are needed to understand better the clinical implications of the findings. Results should be taken into account in the diagnosis of the occlusal development in children with parafunctions to prevent future abnormalities: a bruxist child may have bigger dental arches than a normal child.

Effects of Line and Pillar Array Microengineered SiO2 Thin Films on the Osteogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells

A primary goal in bone tissue engineering is the design of implants that induce controlled, guided, and rapid healing. The events that normally lead to the integration of an implant into bone and determine the performance of the device occur mainly at the tissue-implant interface. Topographical surface modification of a biomaterial might be an efficient tool for inducing stem cell osteogenic differentiation and replace the use of biochemical stimuli. The main goal of this work was to develop micropatterned bioactive silica thin films to induce the osteogenic differentiation of human bone marrow-derived mesenchymal stem cells (hMSCs) only through topographical stimuli. Line and pillar micropatterns were developed by a combination of sol-gel/soft lithography and characterized by scanning electron microscopy, atomic force microscopy, and contact angle measurements. hMSCs were cultured onto the microfabricated thin films and flat control for up to 21 days under basal conditions. The micropatterned groups induced levels of osteogenic differentiation and expression of osteoblast-associated markers higher than those of the flat controls. Via comparison of the micropatterns, the pillars caused a stronger response of the osteogenic differentiation of hMSCs with a higher level of expression of osteoblast-associated markers, ALP activity, and extracellular matrix mineralization after the cells had been cultured for 21 days. These findings suggest that specific microtopographic cues can direct hMSCs toward osteogenic differentiation.

Modulation of human dermal microvascular endothelial cell and human gingival fibroblast behavior by micropatterned silica coating surfaces for zirconia dental implant applications

Abstract Dental ceramic implants have shown superior esthetic behavior and the absence of induced allergic disorders when compared to titanium implants. Zirconia may become a potential candidate to be used as an alternative to titanium dental implants if surface modifications are introduced. In this work, bioactive micropatterned silica coatings were produced on zirconia substrates, using a combined methodology of sol–gel processing and soft lithography. The aim of the work was to compare the in vitro behavior of human gingival fibroblasts (HGFs) and human dermal microvascular endothelial cells (HDMECs) on three types of silica-coated zirconia surfaces: flat and micropatterned (with pillars and with parallel grooves). Our results showed that cells had a higher metabolic activity (HGF, HDMEC) and increased gene expression levels of fibroblast-specific protein-1 (FSP-1) and collagen type I (COL I) on surfaces with pillars. Nevertheless, parallel grooved surfaces were able to guide cell growth. Even capillary tube-like networks of HDMEC were oriented according to the surface geometry. Zirconia and silica with different topographies have shown to be blood compatible and silica coating reduced bacteria adhesion. All together, the results indicated that microstructured bioactive coating seems to be an efficient strategy to improve soft tissue integration on zirconia implants, protecting implants from peri-implant inflammation and improving long-term implant stabilization. This new approach of micropatterned silica coating on zirconia substrates can generate promising novel dental implants, with surfaces that provide physical cues to guide cells and enhance their behavior.

Effect of awareness through movement on the head posture of bruxist children

The aim of this study was to evaluate the effectiveness of physiotherapy to improve the head posture and reduce the signs of bruxism in a group of bruxist children. A single-blind randomized clinical trial was performed. All the subjects were 3- to 6-year old, had complete primary dentition, dental and skeletal class I occlusion and were classified as bruxist according to the minimal criteria of the ICSD for bruxism. For each child, a clinical, photographic and radiographic evaluation of the head and cervical posture were realized with standardized techniques. The children were randomized in an experimental (n = 13) and a control (n = 13) group. A physiotherapeutic intervention was applied to the children of the experimental group once a week, until 10 sessions were completed. Afterwards, the cephalogram and the clinical and photographic evaluation of the head posture were measured again. The data were analysed with the t-test and Mann-Whitney test. The subjects of the experimental group showed statistically significant improvement in the natural head posture. The physiotherapeutic intervention showed to be efficient to improve the head posture at the moment of measurement in the studied children. The relationship between bruxism and head posture, if exists, seems to be worthwhile to examine.

Effects of density of anisotropic microstamped silica thin films on guided bone tissue regeneration--in vitro study.

The growing demand for better implant aesthetics has led to increased research on the development of all-ceramic dental implants. The use of microtextured coatings with enhanced properties has been presented as a viable way to improve tissue integrability of all-ceramic implants. The aim of this study was to evaluate the effects of different densities of anisotropic microtextured silica thin films, which served as a model coating, on the behavior of human osteoblast-like cells. The differential responses of human osteoblast-like cells to anisotropic silica microtextures with varying densities, produced via a combination of sol-gel and soft lithography processing, were evaluated in terms of alignment, elongation (using fluorescence microscopy), overall cellular activity, and the expression/activity levels of alkaline phosphatase (ALP). Statistical analysis was conducted using one-way ANOVA/Tukey HSD post hoc test. The thin films were thoroughly characterized via scanning electron microscopy/energy dispersive spectroscopy, Fourier transform infrared, and contact angle measurements. Thin film characterization revealed increased nanoscale roughness and reduced wettability on the micropatterned surfaces. Cell culture experiments indicated that the microtextures induced cell alignment, elongation, and guided colonization on the surface. Cells cultured on denser micropatterns exhibited increased metabolic activity (t = 14-21 days). The early expression/activity levels of ALP released into the medium were found to be significantly higher only on the least dense micropattern. These results suggest the possibility that microstructured silica thin films could be used to guide and enhance peri-implant cell/tissue responses, potentially improving tissue integration for metallic and all-ceramic dental implants.

Early Spreading and Propagation of Human Bone Marrow Stem Cells on Isotropic and Anisotropic Topographies of Silica Thin Films Produced via Microstamping

While there has been rapid development of microfabrication techniques to produce high-resolution surface modifications on a variety of materials in the last decade, there is still a strong need to produce novel alternatives to induce guided tissue regeneration on dental implants. High-resolution microscopy provides qualitative and quantitative techniques to study cellular guidance in the first stages of cell-material interactions. The purposes of this work were (1) to produce and characterize the surface topography of isotropic and anisotropic microfabricated silica thin films obtained by sol-gel processing, and (2) to compare the in vitro biological behavior of human bone marrow stem cells on these surfaces at early stages of adhesion and propagation. The results confirmed that a microstamping technique can be used to produce isotropic and anisotropic micropatterned silica coatings. Atomic force microscopy analysis was an adequate methodology to study in the same specimen the sintering derived contraction of the microfabricated coatings, using images obtained before and after thermal cycle. Hard micropatterned coatings induced a modulation in the early and late adhesion stages of cell-material and cell-cell interactions in a geometry-dependent manner (i.e., isotropic versus anisotropic), as it was clearly determined, using scanning electron and fluorescence microscopies.

Reinforced Portland cement porous scaffolds for load‐bearing bone tissue engineering applications

Modified Portland cement porous scaffolds with suitable characteristics for load-bearing bone tissue engineering applications were manufactured by combining the particulate leaching and foaming methods. Non-crosslinked polydimethylsiloxane was evaluated as a potential reinforcing material. The scaffolds presented average porosities between 70 and 80% with mean pore sizes ranging from 300 μm up to 5.0 mm. Non-reinforced scaffolds presented compressive strengths and elastic modulus values of 2.6 and 245 MPa, respectively, whereas reinforced scaffolds exhibited 4.2 and 443 MPa, respectively, an increase of ∼62 and 80%. Portland cement scaffolds supported human osteoblast-like cell adhesion, spreading, and propagation (t = 1-28 days). Cell metabolism and alkaline phosphatase activity were found to be enhanced at longer culture intervals (t ≥ 14 days). These results suggest the possibility of obtaining strong and biocompatible scaffolds for bone repair applications from inexpensive, yet technologically advanced materials such as Portland cement.

Cellular Alignment Induction during Early In Vitro Culture Stages Using Micropatterned Glass Coatings Produced by Sol-Gel Process

Cell behaviour such as adhesion, morphology, proliferation and functional activity are highly influenced by surface properties including hydrophobicity, roughness, texture and morphology. These surface properties may be controlled using a mixture of additive coating techniques to produce glass coatings by sol-gel process and soft lithography on dental ceramics. The purpose of this work was to compare cell adhesion and early orientation of Human Bone Marrow (HBM) cells cultured on micro-patterned (micro-PGC) and on flat glass coatings (FGC) produced by sol-gel processing. Spin coating was used to apply SiO2 flat coatings on glass substrates as model surfaces. Photolithography was applied to produce master patterns with microscale dimensions. A moulding technique was used to print micropatterned SiO2 glass coatings produced by a sol-gel process. The coatings were then sintered, sterilized and cultured with HBM cells derived from primary cultures, using a standardized protocol, for 1 and 7 days. Cell morphology and orientation were observed using scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Flat and MPGC with line shaped features were produced. Cells presented a typical osteoblastic morphology on flat surfaces while slimmer, preferentially oriented and more elongated morphologies could be seen on line micro-patterned surfaces. HBM cells cultured on flat glass coatings showed increased tendency to spread and to assume more randomized proliferation when compared to the cells on the micro-patterned glass coatings. Micro-patterned glass coatings showed higher orientation control and smaller delay in the rate of proliferation, in early stages of in vitro culture as compared to flat coatings. These preliminary studies revealed that Micro-PGC induce significant morphological changes and controlled orientation of HBM cells during early stages of cell proliferation.