Micro-computed tomography (micro-CT) evaluation of root canal morphology in immature maxillary third molars.

The endodontic treatment of immature permanent teeth with necrotic pulp is a significant clinical challenge. The success of regenerative endodontic procedure is highly dependent on disinfection of the root canal and an accurate anatomical knowledge of the root canal. The aim of this study was to use micro-computed tomography (micro-CT) analysis to investigate the configuration of root canals in the upper permanent third maxillary molars with incomplete root development in their coronal, apical and middle third portions. Thirty immature third permanent maxillary molars were scanned using a micro-CT system. Then, we measured the diameters and areas of the root canal in the coronal, middle and apical third of the roots. The ratio between the long and short diameter of each root canal was then calculated and the canals were divided into several groups: round, oval, long oval, flat and irregular. The round configuration was not observed in the distobuccal and mesiobuccal roots in any of their anatomical regions. Oval and long oval canals predominated in the distobuccal root. The greatest variations were observed in the mesiobuccal root, with the ribbon-shaped canal predominating in the middle region and an irregular shape in the apical region. In the coronal region of the palatal canal, the round configuration predominated; in the middle third, we observed an almost equivalent distribution between round and oval configurations; apically, the oval shape predominated. In conclusion, we observed significant complexity and variation in the morphology and configuration of root canals in immature permanent molars, thus generating additional obstacles for the success of regenerative endodontics.

Ultra-Short Laser-Assisted Micro-Structure Formations on Mg/Zn Double-Doped Calcium Phosphate Ceramics for Enhanced Antimicrobial Activity.

Bacterial infection is one of the most common and harmful medical issues following the implantation of materials and devices in the body leading to antibiotic resistance of diverse bacterial strains. In this work, a novel approach is presented combining adopted laser-based patterning method in addition to doping (Mg and Zn) metal ions to prepare calcium phosphate ceramic substrate, applicable in medicine, with enhanced surface antimicrobial characteristics. The preparation of tablets containing Mg (8.44 mol%) and Zn (2.63 mol%) ß-tricalcium phosphate involved biomimetic precipitation of amorphous calcium phosphate in media of simulated body fluid enriched with Mg2+ and Zn2+ ions as well as the presence of valine as an organic additive, followed by step-wise calcination up to 1000 °C. The results from laser processing showed formation of deeper patterns with increased surface roughness (from 4.9 µm to 9.4 µm) as laser power and velocity increase, keeping constant the hatch sizes of 50 µm. The textured surfaces consist of peaks and valleys arrangement that change the morphology of Escherichia coli cells and decrease of cell viability. Our study reveals the possibilities of the application of ultra-short laser radiation as a potential alternative therapy for controlling the antimicrobial effect of the ceramic surface.

Design of Surfaces with Persistent Antimicrobial Properties on Stainless Steel Developed Using Femtosecond Laser Texturing for Application in "High Traffic" Objects.

Metal-based high-touch surfaces used for diverse applications in everyday use, like handrails, playground grab handles, doorknobs, ATM touch pads, and desks, are the most common targets for pollution with a variety of microbes; there is thus a need to improve their antimicrobial properties, an issue which has become a challenge in recent years, particularly after the COVID-19 pandemic. According to the World Health Organization (WHO), drug-resistant pathogens are one of the main concerns to global health today, as they lead to longer hospital stays and increased medical costs. Generally, the development of antimicrobial surfaces is related to the utilization of chemical methods via deposition on surfaces in the forms of various types of coatings. However, the addition of chemical substances onto a surface can induce unwanted effects, since it causes surface chemistry changes and, in some cases, cannot provide long-lasting results. A novel approach of utilising ultra-short laser radiation for the treatment of metallic surfaces by inducing a variety of micro- and nanostructuration is elaborated upon in the current research, estimating the optimum relation between the wettability and roughness characteristics for the creation of antimicrobial properties for such high-touch surfaces. In the current study, AISI 304-304L stainless steel metal was used as a benchmark material. Surface texturing via laser ablation with femtosecond laser pulses is an effective method, since it enables the formation of a variety of surface patterns, along with the creation of bimodal roughness, in one-step processing. In this investigation, a precise approach toward developing hydrophobic stainless steel surfaces with tunable adherence using femtosecond laser-induced modification is described. The impact of basic femtosecond laser processing parameters, like the scanning velocity, laser energy, and wettability properties of the laser-processed stainless steel samples, are examined. It is identified that the topography and morphology of laser-induced surface structures can be efficiently changed by adapting the laser processing parameters to create structures, which facilitate the transfer of surface properties from extremely low to high surface wettability.

Single-Step Process for Titanium Surface Micro- and Nano-Structuring and In Situ Silver Nanoparticles Formation by Ultra-Short Laser Patterning.

Ultra-short laser (USL)-induced surface structuring combined with nanoparticles synthesis by multiphoton photoreduction represents a novel single-step approach for commercially pure titanium (cp-Ti) surface enhancement. Such a combination leads to the formation of distinct topographical features covered by nanoparticles. The USL processing of cp-Ti in an aqueous solution of silver nitrate (AgNO3) induces the formation of micron-sized spikes surmounted by silver nanoparticles (AgNPs). The proposed approach combines the structuring and oxidation of the Ti surface and the synthesis of AgNPs in a one-step process, without the use of additional chemicals or a complex apparatus. Such a process is easy to implement, versatile and sustainable compared to alternative methodologies capable of obtaining comparable results. Antimicrobial surfaces on medical devices (e.g., surgical tools or implants), for which titanium is widely used, can be realized due to the simultaneous presence of AgNPs and micro/nano-structured surface topography. The processed surfaces were examined by means of a scanning electron microscope (SEM), energy-dispersive X-ray spectroscopy (EDX), atomic force microscopy (AFM) and Raman spectroscopy. The surface morphology and the oxidation, quality and quantity of AgNPs were analyzed in relation to process parameters (laser scanning speed and AgNO3 concentration), as well as the effect of AgNPs on the Raman signal of Titanium oxide.

Optimizing the Surface Structural and Morphological Properties of Silk Thin Films via Ultra-Short Laser Texturing for Creation of Muscle Cell Matrix Model.

Temporary scaffolds that mimic the extracellular matrix's structure and provide a stable substratum for the natural growth of cells are an innovative trend in the field of tissue engineering. The aim of this study is to obtain and design porous 2D fibroin-based cell matrices by femtosecond laser-induced microstructuring for future applications in muscle tissue engineering. Ultra-fast laser treatment is a non-contact method, which generates controlled porosity-the creation of micro/nanostructures on the surface of the biopolymer that can strongly affect cell behavior, while the control over its surface characteristics has the potential of directing the growth of future muscle tissue in the desired direction. The laser structured 2D thin film matrices from silk were characterized by means of SEM, EDX, AFM, FTIR, Micro-Raman, XRD, and 3D-roughness analyses. A WCA evaluation and initial experiments with murine C2C12 myoblasts cells were also performed. The results show that by varying the laser parameters, a different structuring degree can be achieved through the initial lifting and ejection of the material around the area of laser interaction to generate porous channels with varying widths and depths. The proper optimization of the applied laser parameters can significantly improve the bioactive properties of the investigated 2D model of a muscle cell matrix.

Investigating Potential Effects of Ultra-Short Laser-Textured Porous Poly-ε-Caprolactone Scaffolds on Bacterial Adhesion and Bone Cell Metabolism.

Developing antimicrobial surfaces that combat implant-associated infections while promoting host cell response is a key strategy for improving current therapies for orthopaedic injuries. In this paper, we present the application of ultra-short laser irradiation for patterning the surface of a 3D biodegradable synthetic polymer in order to affect the adhesion and proliferation of bone cells and reject bacterial cells. The surfaces of 3D-printed polycaprolactone (PCL) scaffolds were processed with a femtosecond laser (λ = 800 nm; τ = 130 fs) for the production of patterns resembling microchannels or microprotrusions. MG63 osteoblastic cells, as well as S. aureus and E. coli, were cultured on fs-laser-treated samples. Their attachment, proliferation, and metabolic activity were monitored via colorimetric assays and scanning electron microscopy. The microchannels improved the wettability, stimulating the attachment, spreading, and proliferation of osteoblastic cells. The same topography induced cell-pattern orientation and promoted the expression of alkaline phosphatase in cells growing in an osteogenic medium. The microchannels exerted an inhibitory effect on S. aureus as after 48 h cells appeared shrunk and disrupted. In comparison, E. coli formed an abundant biofilm over both the laser-treated and control samples; however, the film was dense and adhesive on the control PCL but unattached over the microchannels.

Ultra-Short Laser Surface Properties Optimization of Biocompatibility Characteristics of 3D Poly-ε-Caprolactone and Hydroxyapatite Composite Scaffolds.

The use of laser processing for the creation of diverse morphological patterns onto the surface of polymer scaffolds represents a method for overcoming bacterial biofilm formation and inducing enhanced cellular dynamics. We have investigated the influence of ultra-short laser parameters on 3D-printed poly-ε-caprolactone (PCL) and poly-ε-caprolactone/hydroxyapatite (PCL/HA) scaffolds with the aim of creating submicron geometrical features to improve the matrix biocompatibility properties. Specifically, the present research was focused on monitoring the effect of the laser fluence (F) and the number of applied pulses (N) on the morphological, chemical and mechanical properties of the scaffolds. SEM analysis revealed that the femtosecond laser treatment of the scaffolds led to the formation of two distinct surface geometrical patterns, microchannels and single microprotrusions, without triggering collateral damage to the surrounding zones. We found that the microchannel structures favor the hydrophilicity properties. As demonstrated by the computer tomography results, surface roughness of the modified zones increases compared to the non-modified surface, without influencing the mechanical stability of the 3D matrices. The X-ray diffraction analysis confirmed that the laser structuring of the matrices did not lead to a change in the semi-crystalline phase of the PCL. The combinations of two types of geometrical designs-wood pile and snowflake-with laser-induced morphologies in the form of channels and columns are considered for optimizing the conditions for establishing an ideal scaffold, namely, precise dimensional form, mechanical stability, improved cytocompatibility and antibacterial behavior.

Biomimetic Hierarchical Structuring of PLA by Ultra-Short Laser Pulses for Processing of Tissue Engineered Matrices: Study of Cellular and Antibacterial Behavior.

The influence of ultra-short laser modification on the surface morphology and possible chemical alteration of poly-lactic acid (PLA) matrix in respect to the optimization of cellular and antibacterial behavior were investigated in this study. Scanning electron microscopy (SEM) morphological examination of the processed PLA surface showed the formation of diverse hierarchical surface microstructures, generated by irradiation with a range of laser fluences (F) and scanning velocities (V) values. By controlling the laser parameters, diverse surface roughness can be achieved, thus influencing cellular dynamics. This surface feedback can be applied to finely tune and control diverse biomaterial surface properties like wettability, reflectivity, and biomimetics. The triggering of thermal effects, leading to the ejection of material with subsequent solidification and formation of raised rims and 3D-like hollow structures along the processed zones, demonstrated a direct correlation to the wettability of the PLA. A transition from superhydrophobic (θ > 150°) to super hydrophilic (θ < 20°) surfaces can be achieved by the creation of grooves with V = 0.6 mm/s, F = 1.7 J/cm2. The achieved hierarchical architecture affected morphology and thickness of the processed samples which were linked to the nature of ultra-short laser-material interaction effects, namely the precipitation of temperature distribution during material processing can be strongly minimized with ultrashort pulses leading to non-thermal and spatially localized effects that can facilitate volume ablation without collateral thermal damage The obtained modification zones were analyzed employing Fourier transform infrared (FTIR), X-ray photoelectron spectroscopy (XPS), Energy dispersive X-ray analysis (EDX), and optical profilometer. The modification of the PLA surface resulted in an increased roughness value for treatment with lower velocities (V = 0.6 mm/s). Thus, the substrate gains a 3D-like architecture and forms a natural matrix by microprocessing with V = 0.6 mm/s, F = 1.7 J/cm2, and V = 3.8 mm/s, F = 0.8 J/cm2. The tests performed with Mesenchymal stem cells (MSCs) demonstrated that the ultra-short laser surface modification altered the cell orientation and promoted cell growth. The topographical design was tested also for the effectiveness of bacterial attachment concerning chosen parameters for the creation of an array with defined geometrical patterns.

Femtosecond Laser Fabrication of Engineered Functional Surfaces Based on Biodegradable Polymer and Biopolymer/Ceramic Composite Thin Films.

Surface functionalization introduced by precisely-defined surface structures depended on the surface texture and quality. Laser treatment is an advanced, non-contact technique for improving the biomaterials surface characteristics. In this study, femtosecond laser modification was applied to fabricate diverse structures on biodegradable polymer thin films and their ceramic blends. The influences of key laser processing parameters like laser energy and a number of applied laser pulses (N) over laser-treated surfaces were investigated. The modification of surface roughness was determined by atomic force microscopy (AFM). The surface roughness (Rrms) increased from approximately 0.5 to nearly 3 µm. The roughness changed with increasing laser energy and a number of applied laser pulses (N). The induced morphologies with different laser parameters were compared via Scanning electron microscopy (SEM) and confocal microscopy analysis. The chemical composition of exposed surfaces was examined by FTIR, X-ray photoelectron spectroscopy (XPS), and XRD analysis. This work illustrates the capacity of the laser microstructuring method for surface functionalization with possible applications in improvement of cellular attachment and orientation. Cells exhibited an extended shape along laser-modified surface zones compared to non-structured areas and demonstrated parallel alignment to the created structures. We examined laser-material interaction, microstructural outgrowth, and surface-treatment effect. By comparing the experimental results, it can be summarized that considerable processing quality can be obtained with femtosecond laser structuring.