Antibacterial and osteoinductive properties of wollastonite scaffolds impregnated with propolis produced by additive manufacturing.

Biocompatible ceramic scaffolds offer a promising approach to address the challenges in bone reconstruction. Wollastonite, well-known for its exceptional biocompatibility, has attracted significant attention in orthopedics and craniofacial fields. However, the antimicrobial properties of wollastonite have contradictory findings, necessitating further research to enhance its antibacterial characteristics. This study aimed to explore a new approach to improve in vitro biological response in terms of antimicrobial activity and cell proliferation by taking advantage of additive manufacturing for the development of scaffolds with complex geometries by 3D printing using propolis-modified wollastonite. The scaffolds were designed with a TPMS (Triply Periodic Minimal Surface) gyroid geometric shape and 3D printed prior to impregnation with propolis extract. The paste formulation was characterized by rheometric measurements, and the presence of propolis was confirmed by FTIR spectroscopy. The scaffolds were comprehensively assessed for their mechanical strength. The biological characterization involved evaluating the antimicrobial effects against Staphylococcus aureus and Staphylococcus epidermidis, employing Minimum Inhibitory Concentration (MIC), Zone of Inhibition (ZOI), and biofilm formation assays. Additionally, SaOs-2 cultures were used to study cell proliferation (Alamar blue assay), and potential osteogenic was tested (von Kossa, Alizarin Red, and ALP stainings) at different time points. Propolis impregnation did not compromise the mechanical properties of the scaffolds, which exhibited values comparable to human trabecular bone. Propolis incorporation conferred antibacterial activity against both Staphylococcus aureus and Staphylococcus epidermidis. The implementation of TPMS gyroid geometry in the scaffold design demonstrated favorable cell proliferation with increased metabolic activity and osteogenic potential after 21 days of cell cultures.

Cyclic strain of poly (methyl methacrylate) surfaces triggered the pathogenicity of Candida albicans.

Candida albicans is an opportunistic yeast and the primary etiological factor in oral candidiasis and denture stomatitis. The pathogenesis of C. albicans could be triggered by several variables, including environmental, nutritional, and biomaterial surface cues. Specifically, biomaterial interactions are driven by different surface properties, including wettability, stiffness, and roughness. Dental biomaterials experience repetitive (cyclic) stresses from chewing and biomechanical movements. Pathogenic biofilms are formed over these biomaterial surfaces under cyclic strain. This study investigated the effect of the cyclic strain (deformation) of biomaterial surfaces on the virulence of Candida albicans. Candida biofilms were grown over Poly (methyl methacrylate) (PMMA) surfaces subjected to static (no strain) and cyclic strain with different levels (ε˜x=0.1 and 0.2%). To evaluate the biomaterial-biofilm interactions, the biofilm characteristics, yeast-to-hyphae transition, and the expression of virulent genes were measured. Results showed the biofilm biomass and metabolic activity to be significantly higher when Candida adhered to surfaces subjected to cyclic strain compared to static surfaces. Examination of the yeast-to-hyphae transition showed pseudo-hyphae cells (pathogenic) in cyclically strained biomaterial surfaces, whereas static surfaces showed spherical yeast cells (commensal). RNA sequencing was used to determine and compare the transcriptome profiles of cyclically strained and static surfaces. Genes and transcription factors associated with cell adhesion (CSH1, PGA10, and RBT5), biofilm formation (EFG1), and secretion of extracellular matrix (ECM) (CRH1, ADH5, GCA1, and GCA2) were significantly upregulated in the cyclically strained biomaterial surfaces compared to static ones. Genes and transcription factors associated with virulence (UME6 and HGC1) and the secretion of extracellular enzymes (LIP, PLB, and SAP families) were also significantly upregulated in the cyclically strained biomaterial surfaces compared to static. For the first time, this study reveals a biomaterial surface factor triggering the pathogenesis of Candida albicans, which is essential for understanding, controlling, and preventing oral infections. STATEMENT OF SIGNIFICANCE: Fungal infections produced by Candida albicans are a significant contributor to various health conditions. Candida becomes pathogenic when certain environmental conditions change, including temperature, pH, nutrients, and CO2 levels. In addition, surface properties, including wettability, stiffness, and roughness, drive the interactions between Candida and biomaterials. Clinically, Candida adheres to biomaterials that are under repetitive deformation due to body movements. In this work, we revealed that when Candida adhered to biomaterial surfaces subjected to repetitive deformation, the microorganism becomes pathogenic by increasing the formation of biofilms and the expression of virulent factors related to hyphae formation and secretion of enzymes. Findings from this work could aid the development of new strategies for treating fungal infections in medical devices or implanted biomaterials.

Effects of Propolis Impregnation on Polylactic Acid (PLA) Scaffolds Loaded with Wollastonite Particles against Staphylococcus aureus, Staphylococcus epidermidis, and Their Coculture for Potential Medical Devices.

Several diseases and injuries cause irreversible damage to bone tissues, which may require partial or total regeneration or replacement. Tissue engineering suggests developing substitutes that may contribute to the repair or regeneration process by using three-dimensional lattices (scaffolds) to create functional bone tissues. Herein, scaffolds comprising polylactic acid and wollastonite particles enriched with propolis extracts from the Arauca region of Colombia were developed as gyroid triply periodic minimal surfaces using fused deposition modeling. The propolis extracts exhibited antibacterial activity against Staphylococcus aureus (ATCC 25175) and Staphylococcus epidermidis (ATCC 12228), which cause osteomyelitis. The scaffolds were characterized using scanning electron microscopy, Fourier-transform infrared spectroscopy, differential scanning calorimetry, contact angle, swelling, and degradation. Their mechanical properties were assessed using static and dynamic tests. Cell viability/proliferation assay was conducted using hDP-MSC cultures, while their bactericidal properties against monospecies cultures (S. aureus and S. epidermidis) and cocultures were evaluated. The wollastonite particles did not affect the physical, mechanical, or thermal properties of the scaffolds. The contact angle results showed that there were no substantial differences in the hydrophobicity between scaffolds with and without particles. Scaffolds containing wollastonite particles suffered less degradation than those produced using PLA alone. A representative result of the cyclic tests at Fmax = 450 N showed that the maximum strain reached after 8000 cycles is well below the yield strain (i.e., <7.5%), thereby indicating that even under these stringent conditions, these scaffolds will be able to work properly. The scaffolds impregnated with propolis showed a lower % of cell viability using hDP-MSCs on the 3rd day, but these values increased on the 7th day. These scaffolds exhibited antibacterial activity against the monospecies cultures of S. aureus and S. epidermidis and their cocultures. The samples without propolis loads did not show inhibition halos, whereas those loaded with EEP exhibited halos of 17.42 ± 0.2 mm against S. aureus and 12.9 ± 0.5 mm against S. epidermidis. These results made the scaffolds possible bone substitutes that exert control over species with a proliferative capacity for the biofilm-formation processes required for typical severe infectious processes.

Preparation of collagen fibrils from mineralized tissues and evaluation by atomic force microscopy.

Mineralized tissues like bone and dentin are materials that support the distribution of mechanical loads through the body of humans and other animals. While their organic content plays a critical role on the structural behavior of these materials, investigations that quantify the structural properties of collagen fibrils in mineralized tissues at the nanoscale are rather limited. We report a new experimental methodology to prepare samples of dentinal collagen fibrils for evaluation by atomic force microscopy and characterize their mechanical behavior. Specifically, a Dynamic Mechanical Analysis (DMA) of the collagen fibrils was performed to study their viscoelastic behavior. The capacity for viscous dampening in the fibrils was characterized in terms of measures of the energy dissipation, phase angle and loss modulus in both the peak and trough regions of the fibrils. According to the phase angle and the loss modulus, the peak regions of the fibrils exhibit significantly greater stiffness and capacity for dampening than the trough regions. This new approach will help in exploring the role of collagen fibrils in the mechanical behavior of dentin and other mineralized tissues as well as help to understand the potential effects from changes in fibril confirmation with tissue treatments, aging or that result from chronic disease.

Bioinspired hierarchical impact tolerant materials.

The quest for new light-weight materials with superior mechanical properties is a goal of materials scientists and engineers worldwide. A promising route in this pursuit is drawing inspiration from nature to design and develop materials with enhanced properties. By emulating the graded mineral content and hierarchical structure of fish scales of the Arapaima gigas from the nano to macro scales, we were able to develop bioinspired laminated composites with improved impact resistance. Activated by the addition of nano-particles of Al2O3 and nano-layers of TiN to a thermoplastic fiber substrate, new energy dissipation mechanisms operating at the nanoscale enhanced the energy absorption and stiffness of the bioinspired material. Remarkably, the newly developed materials are easily transferred to the industry with minimum associated manufacturing costs.

Contribution of Root Canal Treatment to the Fracture Resistance of Dentin.

INTRODUCTION: Although the strength and toughness of dentin decrease with age, no study has explored if restorative treatments are a contributing factor. METHODS: Multiple extracted teeth were obtained from randomly selected donors and categorized according to donor age and prior root canal treatment. The microstructure and chemical composition of radicular dentin were evaluated using scanning electron microscopy and Raman spectroscopy, respectively, and the strength was evaluated in 4-point flexure to failure. Data were compared using the Student t test. RESULTS: Dentin from the root canal-restored teeth exhibited significantly lower strength (P < .05) than tissue from age- and donor-matched unrestored tooth pairs. Although there was no significant difference in the mineral-to-collagen ratio between the 2 groups, dentin obtained from the root canal-treated teeth exhibited more extensive collagen cross-linking and lower tubule occlusion ratios than the unrestored tooth pairs. CONCLUSIONS: There is a decrease in the strength of radicular dentin with aging, but prior root canal treatment increases the extent of degradation.

Reduction in Fracture Resistance of the Root with Aging.

INTRODUCTION: The incidence of vertical root fracture in endodontically treated teeth increases with patient age. This study evaluated the microstructure, chemical composition, and mechanical properties of radicular dentin as a function of aging. METHODS: Single-rooted teeth were obtained from adult donors ranging from age 15 to older than 80 years. Beams were extracted from the middle third of the root and subjected to 4-point flexure to failure to evaluate strength of dentin in terms of donor age. Based on the strength distribution, the fatigue strength of root tissue from young (≤30 years) and old (≥55 years) donor teeth was evaluated via cyclic flexure loading. The microstructure and chemical composition of radicular dentin from the 2 groups were evaluated as a function of distance from the root apex using microscopy and Raman spectroscopy, respectively. RESULTS: The strength decreased with age by approximately 25 MPa per decade, which resulted in a significant difference (P ≤ .0001) between the young (199 ± 36 MPa) and old (122 ± 11 MPa) groups. There was also a significant difference (P ≤ .0001) in fatigue strength between the young and old age groups. Both the mineral-to-collagen ratio and degree of cross-linking increased with age, with the largest changes identified in the apical and middle thirds, respectively. CONCLUSIONS: There is a reduction in the damage tolerance of radicular dentin with increasing age. This degradation appears to result from changes in the microstructure, corresponding chemical composition, and increase in collagen cross-linking with age, and is most severe near the root apex.

Effects of polar solvents on the mechanical behavior of fish scales.

Fish scales are unique structural materials that serve as a form of natural armor. In this investigation the mechanical behavior of scales from the Cyprinus carpio was evaluated after exposure to a polar solvent. Uniaxial tensile and tear tests were conducted on specimens prepared from the scales of multiple fish extracted from near the head, middle and tail regions, and after exposure to ethanol for periods from 0 to 24h. Submersion in ethanol caused instantaneous changes in the tensile properties regardless of anatomical site, with increases in the elastic modulus, strength and modulus of toughness exceeding 100%. The largest increase in properties overall occurred in the elastic modulus of scales from the tail region and exceeded 200%. Although ethanol treatment had significant effect on the tensile properties, it had limited influence on the tear resistance. The contribution of ethanol to the mechanical behavior appears to be derived from an increase in the degree of interpeptide hydrogen-bonding of the collagen molecules. Spatial variations in the effects of ethanol exposure on the mechanical behavior arise from the differences in degree of mineralization and lower mineral content in scales of the tail region.

Temperature effects on the fracture resistance of scales from Cyprinus carpio.

In this investigation the fracture resistance of scales from Cyprinus carpio was evaluated as a function of environmental temperature. Tear specimens were prepared from scales obtained from three characteristic regions (i.e. head, mid-length and tail) of multiple fish. The fracture resistance was characterized in Mode III loading and over temperatures ranging from -150°C to 21°C. Results showed that there was a significant reduction in tear resistance with decreasing temperature and the lowest resistance to fracture was obtained at -150°C. There was a significant difference in the relative tear toughness between scales from the three locations at ambient conditions (21°C), but not below freezing. Scales obtained near the head exhibited the largest resistance to fracture (energy ≈ 150 ± 25 kJm(-2)) overall. The fracture resistance was found to be primarily dependent on the thickness of the external mineralized layer and the number of external elasmodine plies, indicating that both the anatomical position and the corresponding microstructure are important to the mechanical behavior of elasmoid fish scales. These variables may be exploited in the design of bioinspired armors and should be considered in future studies concerning the mechanical behavior of these interesting natural materials.