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For decades, animal models have been the standard approach in drug research and development, as they are required by regulations in the transition from preclinical to clinical trials. However, there is growing ethical and scientific concern regarding these trials, as 80 % of the therapeutic potential observed in pre-clinical studies are often unable to be replicated, despite demonstrating efficacy and safety. In response to this, Tissue Engineering has emerged as a promising alternative that enables the treatment of various diseases through the production of biological models for advanced biological assays or through the direct development of tissue repairs or replacements. One of the promising applications of Tissue Engineering is the development of three-dimensional (3D) models for in vitro tests, replacing the need for in vivo animal models. In this study, 3D skin equivalents (TSE) were produced and used as an in vitro model to test photobiostimulation using curcumin-loaded nanocapsules. Photodynamic biostimulation therapy uses photodynamic processes to generate small amounts of reactive oxygen species (ROS), which can activate important biological effects such as cell differentiation, modulation of inflammatory processes and contribution to cell regeneration. The PLGA nanocapsules (NC) used in the study were synthesized through a preformed polymer deposition method, exhibiting particle size <200 nm, Zeta potential >|30| and polydispersity index between 0.5 and 0.3. Atomic force microscopy analyzes confirmed that the particle size was <200 nm, with a spherical morphology and a predominantly smooth and uniform surface. The NC biocompatibility assay did not demonstrate cytotoxicity for the concentrations tested (2.5-25 µg mL-1).The in vitro release assay showed a slow and sustained release characteristic of the nanocapsules, and cellular uptake assays indicated a significant increase in cellular internalization of the curcumin-loaded nanostructure. Monolayer photobiostimulation studies revealed an increase in cell viability of the HDFn cell line (viability 134 %-228 %) for all LED fluences employed at λ = 450 nm (150, 300, and 450 mJ cm-2). Additionally, the scratch assays, monitoring in vitro scar injury, demonstrated more effective effects on cell proliferation with the fluence of 300 mJ cm-2. Staining of TSE with hematoxylin and eosin showed the presence of cells with different morphologies, confirming the presence of fibroblasts and keratinocytes. Immunohistochemistry using KI-67 revealed the presence of proliferating cells in TSE after irradiation with LED λ = 450 nm (150, 300, and 450 mJ cm-2).

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BACKGROUND: The facial aging process entails alterations in the volume, shape, and texture of all skin layers over time. Calcium hydroxyapatite (CaHA) is a well-established safe skin filler with unique properties to resolve some skin alterations by stimulating neocollagenesis. The vectoral-lift (V-lift) technique targets the global repositioning of facial structures by addressing distinct anatomical injection planes. It includes deep facial augmentation with Radiesse PlusTM to retain ligament restructuring and superficial subcutaneous enhancement with diluted Radiesse DuoTM. Herein, we present cases that illustrate the use of this approach. METHODS: This pilot study enrolled 36 participants (33 women and three men; ages 37-68 years) in a Brazilian clinical setting, and all patients underwent a single treatment. Photographs were taken at rest, in frontal and oblique views, before injection, and 90 days after treatment. RESULTS: Treatment resulted in elevation of the upper and middle face, notable improvements in the infraorbital hollow, and adjustment of the mean facial volume. CONCLUSIONS: The V-lift technique is a three-dimensional pan-facial treatment that relies on ligament support and face vectoring to obtain a lifting effect and facial contour restoration. It encompasses deep facial augmentation involving the use of Radiesse PlusTM for restructuring and retaining ligaments and Radiesse DuoTM for superficial subcutaneous enhancement. This approach targets a global repositioning of the facial structures by addressing distinct anatomical injection planes. It achieves a repositioning of the overall facial anatomy without requiring a substantial volumetric expansion. LEVEL OF EVIDENCE IV: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

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The use of biofuels has grown in the last decades as a consequence of the direct environmental impacts of fossil fuel use. Elucidating structure, diversity, species interactions, and assembly mechanisms of microbiomes is crucial for understanding the influence of environmental disturbances. However, little is known about how contamination with biofuel/petrofuel blends alters the soil microbiome. Here, we studied the dynamics in the soil microbiome structure and composition of four field areas under long-term contamination with biofuel/fossil fuel blends (ethanol 10% and gasoline 90%-E10; ethanol 25% and gasoline 75%-E25; soybean biodiesel 20% and diesel 80%-B20) submitted to different bioremediation treatments along a temporal gradient. Soil microbiomes from biodiesel-polluted areas exhibited higher richness and diversity index values and more complex microbial communities than ethanol-polluted areas. Additionally, monitored natural attenuation B20-polluted areas were less affected by perturbations caused by bioremediation treatments. As a consequence, once biostimulation was applied, the degradation was slower compared with areas previously actively treated. In soils with low diversity and richness, the impact of bioremediation treatments on the microbiomes was greater, and as a result, the hydrocarbon degradation extent was higher. The network analysis showed that all abundant keystone taxa corresponded to well-known degraders, suggesting that the abundant species are core targets for biostimulation in soil remediation processes. Altogether, these findings showed that the knowledge gained through the study of microbiomes in contaminated areas may help design and conduct optimized bioremediation approaches, paving the way for future rationalized and efficient pollutant mitigation strategies.

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In this work, the bioremediation of wastewater from the textile industry with indigo dye content was carried out using combined bioaugmentation, bioventilation, and biostimulation techniques. Initially, the inoculum was prepared by isolating the microorganisms from the textile wastewater in a 2 L bioreactor. Then, the respirometry technique was implemented to determine the affinity of the microorganisms and the substrate by measuring CO2 and allowed the formulation of an empirical mathematical model for the growth kinetics of the microorganism. Finally, the bioremediation was carried out in a 3 L bioreactor obtaining an indigo dye removal efficiency of 20.7 ± 1.2%, 24.0 ± 1.5%, and 29.7 ± 1.1% for equivalent wavelengths of 436 nm, 525 nm, and 620 nm. The chemical oxygen demand showed an average reduction of 88.9 ± 2.5%, going from 470.7 ± 15.6 to 52.3 ± 10.7 ppm after 30 days under constant agitation and aeration. A negative generalized exponential model was fitted to assess the affinity of the microorganism with the wastewater as a substrate by evaluating the production of CO2 during the bioremediation. Bioremediation techniques improve water discharge parameters compared to chemical treatments implemented in the industry, reducing the use of substances that can generate secondary pollution. Bioaugmentation, biostimulation, and bioventing of the textile wastewater in this study demonstrate the potential of these combined techniques to serve as an efficient alternative for indigo-contaminated wastewater in the textile industry.

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The aim of this study was to evaluate the effectiveness of photobiomodulation with a 780 nm laser as an adjunct to surgical treatment in the regeneration of bone fractures. Twenty patients diagnosed with open fractures in the lower limbs were selected and randomly divided into two groups: control and LLLT. LLLT parameter: 780 nm, 0.04 cm2 of light beam diameter, 40 mW of power, 10 s per point, 0.4 J of energy, fluence of 10 J/cm2 and irradiance of 1 W/cm2. The evaluated data were: pain, using McGill scale, use of analgesics and anti-inflammatories, levels of cytokines TNF-α, IFN-γ, IL-1ß, IL-10, and IL-17, and bone level regeneration. Data were analyzed using Wilcoxon and Mann-Whitney tests (5%). We can conclude that LLLT was effective as an adjuvant in the bone fracture regeneration process, altered IL-1ß levels, reduced the use of analgesics and anti-inflammatories, reducing the pain pattern throughout the sessions.

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Soil salinization negatively impacts plant development and induces land degradation, thus affecting biodiversity, water quality, crop production, farmers' well-being, and the economic situation in the affected region. Plant germination, growth, and productivity are vital processes impaired by salinity stress; thus, it is considered a serious threat to agriculture. The extent to which a plant is affected by salinity depends mainly on the species, but other factors, including soil attributes, water, and climatic conditions, also affect a plant's ability to tolerate salinity stress. Unfortunately, this phenomenon is expected to be exacerbated further by climate change. Consequently, studies on salt stress tolerance in plants represent an important theme for the present Special Issue of Plants. The present Special Issue contains 14 original contributions that have documented novel discoveries regarding induced or natural variations in plant genotypes to cope with salt stress, including molecular biology, biochemistry, physiology, genetics, cell biology, modern omics, and bioinformatic approaches. This Special Issue also includes the impact of biostimulants on the biochemical, physiological, and molecular mechanisms of plants to deal with salt stress and on the effects of salinity on plant nutrient status. We expect that readers and academia will benefit from all the articles included in this Special Issue.

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The present study describes practical implication of bioaugmentation and biostimulation processes for bioremediation of an industrial soil chronically contaminated by hydrocarbons. For this purpose, biomass production of six autochthonous hydrocarbon-degrading bacteria were evaluated as inoculum of bioaugmentation strategy, by testing carbon and nitrogen sources included co-products and agro-industrial waste as sustainable and low-cost components of the growth medium. Otherwise, biostimulation was approached by the addition of optimized concentration of nitrogen and phosphorus. Microcosm assays showed that total hydrocarbons (TH) were significantly removed from chronically contaminated soil undergoing bioremediation treatment. Systems Mix (bioaugmentation); N,P (biostimulation) and Mix + N,P (bioaugmentation and biostimulation) reached higher TH removal, being 89.85%, 91.00%, 93.04%, respectively, comparing to 77.83% of system C (natural attenuation) at 90 days. The increased heterotrophic aerobic bacteria and hydrocarbon degrading bacteria counts were according to TH biodegrading process during the experiments. Our results showed that biostimulation with nutrients represent a valuable alternative tool to treat a chronically hydrocarbon-contaminated industrial soil, while bioaugmentation with a consortium of hydrocarbon degrading bacteria would be justified when the soil has a low amount of endogenous degrading microorganisms. Furthermore, the production of inoculum for application in bioaugmentation using low-cost substrates, such as industrial waste, would lead to the development of an environmentally friendly and attractive process in terms of cost-benefit.

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Contamination of soils by automotive residual oil represents a global environmental problem. Bioremediation is the technology most suitable to remove this contaminant from the medium. Therefore, this work aimed to evaluate the effectiveness of bioremediation of automotive residual oil-contaminated soils by biostimulation with enzymes, surfactant, and vermicompost. The bioremediation efficiency was examined using a factorial design of 24 to determine the effect of the time, pH and temperature conditions, biostimulation with enzyme-vermicompost, and biostimulation with enzyme-surfactant. Enzymes obtained from Ricinus communis L. seeds, commercial vermicompost, and Triton X-100 were used. Results showed that the highest removal efficiency (99.9%) was achieved at 49 days, with a pH of 4.5, temperature of 37 °C, and using biostimulation with enzyme-vermicompost (3% w/v-5% w/w). The addition of surfactant was not significant in increasing the removal efficiency. Therefore, the results provide adequate conditions to bioremediate automotive residual oil-contaminated soils by biostimulation using enzymes supported with vermicompost.

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This study aims at the application of a marine fungal consortium (Aspergillus sclerotiorum CRM 348 and Cryptococcus laurentii CRM 707) for the bioremediation of diesel oil-contaminated soil under microcosm conditions. The impact of biostimulation (BS) and/or bioaugmentation (BA) treatments on diesel-oil biodegradation, soil quality, and the structure of the microbial community were studied. The use of the fungal consortium together with nutrients (BA/BS) resulted in a TPH (Total Petroleum Hydrocarbon) degradation 42% higher than that obtained by natural attenuation (NA) within 120 days. For the same period, a 72 to 92% removal of short-chain alkanes (C12 to C19) was obtained by BA/BS, while only 3 to 65% removal was achieved by NA. BA/BS also showed high degradation efficiency of long-chain alkanes (C20 to C24) at 120 days, reaching 90 and 92% of degradation of icosane and heneicosane, respectively. In contrast, an increase in the levels of cyclosiloxanes (characterized as bacterial bioemulsifiers and biosurfactants) was observed in the soil treated by the consortium. Conversely, the NA presented a maximum of 37% of degradation of these alkane fractions. The 5-ringed PAH benzo(a)pyrene, was removed significantly better with the BA/BS treatment than with the NA (48 vs. 38 % of biodegradation, respectively). Metabarcoding analysis revealed that BA/BS caused a decrease in the soil microbial diversity with a concomitant increase in the abundance of specific microbial groups, including hydrocarbon-degrading (bacteria and fungi) and also an enhancement in soil microbial activity. Our results highlight the great potential of this consortium for soil treatment after diesel spills, as well as the relevance of the massive sequencing, enzymatic, microbiological and GC-HRMS analyses for a better understanding of diesel bioremediation.

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The addax antelope (Addax nasomaculatus) is a species under serious threat of extinction, as it is more abundant in captivity than in the wild. However, little is known about its basic biology. The aims of this study were to determine how locomotor, feeding, aggressive, marking, and sexual behavior of male addax allocated in all-male groups vary with season and with female contact (i.e., biostimulation). The study was conducted in captive conditions, in two groups of adult males: one with no-physical contact with females, aside from visual and olfactory interactions (CF group, n = 4), and another group completely isolated from females (IF group, n = 4). The frequency of behaviors was recorded during the daytime, 4 days per season (total time of observation = 256 h). Lying, standing, walking, aggressive, marking, grazing, and ruminating behaviors as well as water and supplement consumptions varied with season (all p < 0.05). The lying, walking, marking, grazing, and ruminating behaviors were more frequently observed for CF than IF males (all p < 0.05). Also, all behaviors, except for marking, varied with the interaction between the group and seasons (all p < 0.05). Sexual behavior was extremely scarce, so it was not possible to analyze how it varied with seasons and the group. The present study suggests that management program and housing conditions, especially in ex situ breeding plans, should consider the influence of the season and the sociosexual context on the behavior of addax males.

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Lysinibacillus is a bacterial genus that has generated recent interest for its biotechnological potential in agriculture. Strains belonging to this group are recognized for their mosquitocidal and bioremediation activity. However, in recent years some reports indicate its importance as plant growth promoting rhizobacteria (PGPR). This research sought to provide evidence of the PGP activity of Lysinibacillus spp. and the role of the indole-3-acetic acid (IAA) production associated with this activity. Twelve Lysinibacillus spp. strains were evaluated under greenhouse conditions, six of which increased the biomass and root architecture of corn plants. In most cases, growth stimulation was evident at 108 CFU/mL inoculum concentration. All strains produced IAA with high variation between them (20-70 µg/mL). The bioinformatic identification of predicted genes associated with IAA production allowed the detection of the indole pyruvic acid pathway to synthesize IAA in all strains; additionally, genes for a tryptamine pathway were detected in two strains. Extracellular filtrates from all strain's cultures increased the corn coleoptile length in an IAA-similar concentration pattern, which demonstrates the filtrates had an auxin-like effect on plant tissue. Five of the six strains that previously showed PGPR activity in corn also promoted the growth of Arabidopsis thaliana (col 0). These strains induced changes in root architecture of Arabidopsis mutant plants (aux1-7/axr4-2), the partial reversion of mutant phenotype indicated the role of IAA on plant growth. This work provided solid evidence of the association of Lysinibacillus spp. IAA production with their PGP activity, which constitutes a new approach for this genus. These elements contribute to the biotechnological exploration of this bacterial genus for agricultural biotechnology.

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A well-defined jawline is aesthetically pleasing, so that rejuvenation of the jawline is becoming part of routine aesthetic practice. Restoring balance to the lower face without surgical intervention requires a multiple treatment approach, among which the minimally invasive injectables play a central role. Nevertheless, amidst the plethora of different injectable products available, choosing the option that best suits the patient's need can be a challenge. A panel of experts sought to describe herein 4 clinical cases, illustrating the Contouring Plus technique, which provides a practical guidance for lower third assessment, choice of products, and treatment execution, especially among hyaluronic acid and calcium hydroxylapatite, aiming for immediate, as well as mid-to-long-term results.

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Anthropogenic activities have been increasing Polycyclic Aromatic Hydrocarbons (PAHs) release, promoting an urgent need for decontamination methods. Therefore, anthracene biodegradation by endophytic, extremophilic, and entomophilic fungi was studied. Moreover, a salting-out extraction methodology with the renewable solvent ethanol and the innocuous salt K2HPO4 was employed. Nine of the ten employed strains biodegraded anthracene in liquid medium (19-56% biodegradation) after 14 days at 30 °C, 130 rpm, and 100 mg L-1. The most efficient strain Didymellaceae sp. LaBioMMi 155, an entomophilic strain, was employed for optimized biodegradation, aiming at a better understanding of how factors like pollutant initial concentration, pH, and temperature affected this process. Biodegradation reached 90 ± 11% at 22 °C, pH 9.0, and 50 mg L-1. Futhermore, 8 different PAHs were biodegraded and metabolites were identified. Then, experiments with anthracene in soil ex situ were performed and bioaugmentation with Didymellaceae sp. LaBioMMi 155 presented better results than natural attenuation by the native microbiome and biostimulation by the addition of liquid nutrient medium into soil. Therefore, an expanded knowledge about PAHs biodegradation processes was achieved with emphasis to the action of Didymellaceae sp. LaBioMMi 155, which can be further employed for in situ biodegradation (after strain security test), or for enzyme identification and isolation aiming at oxygenases with optimal activity under alkaline conditions.

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Agriculture in the current century is seeking sustainable tools in order to generate plant production systems with minimal negative environmental impact. In recent years it has been shown that the use of insect frass is an option to be used for this purpose. The present work studied the effect of low doses (0.1, 0.5, and 1.0% w/w) of cricket frass (Acheta domesticus) in the substrate during the cultivation of tomatos under greenhouse conditions. Plant performance and antioxidant enzymatic activities were measured in the study as explicative variables related to plant stress responses in order to determine possible biostimulant or elicitor effects of cricket frass treatments during tomato cultivation under greenhouse conditions. The main findings of this study indicated that tomato plants responded in a dose dependent manner to cricket frass treatments, recalling the hormesis phenomenon. On the one hand, a 0.1% (w/w) cricket frass treatment showed typical biostimulant features, while on the other hand, 0.5 and 1.0% treatments displayed elicitor effects in tomato plants under evaluated conditions in the present study. These results support the possibility that low doses of cricket frass might be used in tomato cultivation (and perhaps in other crops) for biostimulant/elicitor input into sustainable production systems.

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Oily sludge is a residue from the petroleum industry composed of a mixture of sand, water, metals, and high content of hydrocarbons (HCs). The heavy oily sludge used in this study originated from Colombian crude oil with high density and low American Petroleum Institute (API) gravity. The residual waste from heavy oil processing was subject to thermal and centrifugal extraction, resulting in heavy oily sludge with very high density and viscosity. Biodegradation of the total petroleum hydrocarbons (TPH) was tested in microcosms using several bioremediation approaches, including: biostimulation with bulking agents and nutrients, the surfactant Tween 80, and bioaugmentation. Select HC degrading bacteria were isolated based on their ability to grow and produce clear zones on different HCs. Degradation of TPH in the microcosms was monitored gravimetrically and with gas chromatography (GC). The TPH removal in all treatments ranged between 2 and 67%, regardless of the addition of microbial consortiums, amendments, or surfactants within the tested parameters. The results of this study demonstrated that bioremediation of heavy oily sludge presents greater challenges to achieve regulatory requirements. Additional physicochemical treatments analysis to remediate this recalcitrant material may be required to achieve a desirable degradation rate.

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This study evaluated the effect of the application of mineral fertilization (F), the anionic surfactant Triton X-100 (TX100), or the inoculation with a hydrocarbooclastic bacterial consortium (BCons) on the growth of Clitoria ternatea during the phytoremediation of a Gleysol contaminated with weathered petroleum hydrocarbons (39,000 mg kg-1 WPH) collected from La Venta, Tabasco (Mexico). The experiment consisted of a completely randomized design with seven treatments and four replications each under greenhouse conditions. The application of F (biostimulation) increased plant growth and biomass production; in contrast, TX100 only favored root biomass (11%) but significantly favored WPH degradation. Bioaugmentation with BCons did not show significant effects on plant growth. Nevertheless, the combination of biostimulation with bioaugmentation (BCons + F, BCons + TX100, and BCons + F+TX100) enhanced plant growth, hydrocarbonoclastic bacteria population, and WPH degradation when compared to treatments with the single application of bioaugmentation (BCons) or biostimulation (F).

Application of mineral fertilization and commercial surfactant favored root biomass and degradation of weathered petroleum hydrocarbons (WPH). The reintroduction of hydrocarbonoclastic and surfactant-producer bacteria did not enhance plant growth but significantly contributed on WPH degradation from a chronically contaminated soil.

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Currently, the use of biostimulants in agriculture is a tool for mitigating certain environmental stresses. Brown algae extracts have become one of the most important categories of biostimulants in agriculture, and are derived from the different uses and positive results obtained under optimal and stressful conditions. This study aimed to examine the efficacy of a foliar application of a hydroalcoholic extract of Sargassum spp. and two controls (a commercial product based on Ascophyllum nodosum and distilled water) with regard to growth, the antioxidant system, and the expression of defense genes in tomato seedlings grown in nonsaline (0 mM NaCl) and saline (100 mM NaCl) conditions. In general, the results show that the Sargassum extract increased the growth of the seedlings at the end of the experiment (7.80%) compared to the control; however, under saline conditions, it did not modify the growth. The Sargassum extract increased the diameter of the stem at the end of the experiment in unstressed conditions by 14.85% compared to its control and in stressful conditions by 16.04% compared to its control. Regarding the accumulation of total fresh biomass under unstressed conditions, the Sargassum extract increased it by 19.25% compared to its control, and the accumulation of total dry biomass increased it by 18.11% compared to its control. Under saline conditions, the total of fresh and dry biomass did not change. Enzymatic and nonenzymatic antioxidants increased with NaCl stress and the application of algal products (Sargassum and A. nodosum), which was positively related to the expression of the defense genes evaluated. Our results indicate that the use of the hydroalcoholic extract of Sargassum spp. modulated different physiological, metabolic, and molecular processes in tomato seedlings, with possible synergistic effects that increased tolerance to salinity.

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Global climate change affects agriculture and tends to aggravate the effect of various environmental stress factors including soil salinity. Beneficial elements such as titanium (Ti) may improve the performance of plants facing restrictive environments such as saline soils. This research work evaluated the individual effect of sodium chloride (0, 50, and 100 mM NaCl) in solution, that of leaf-applied Ti (0, 500, and 1000 mg L-1 Ti), and their interactions on physiological, biochemical, and nutritional variables of tomato (Solanum lycopersicum L.) seedlings cv. Rio Grande in a factorial design in greenhouse hydroponics. NaCl reduced seedling height, stem diameter, leaf area, SPAD units, and sugar and K concentrations, and increased antioxidant activity in stems and roots, photosynthetic pigments, sugars. Titanium increased the N, P, K, Ca, Mg, and Ti concentrations in leaves, but the concentration of total sugars in leaves was reduced when applying 500 mg Ti L-1. Under moderate salinity conditions (50 mM NaCl) the application of Ti increased the antioxidant activity in roots, while, at all salinity levels tested, Ti increased the concentrations of macro-nutrients and Ti in leaves. Titanium is concluded to have a positive effect on the antioxidant activity and nutrition of seedlings under saline stress conditions.

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Current agricultural practices for vegetable production are unsustainable, and the use of certain nanomaterials has shown significant potential for either plant growth promotion or defense induction in crop species. The aim of the present work was to evaluate the possible effects of two SBA nano-structured silica materials differing in morphology; SBA-15, with porous structure in parallel and with a highly ordered hexagonal array and SBA-16, with spheric nano-cages located in cubic arrays, as plant growth promoters/eustressors on chili pepper (Capsicum annuum L.) during cultivation under greenhouse conditions. The study was carried out at three foliarly applied concentrations (20, 50 and 100 ppm) of either SBA materials to determine effects on seed germination, seedling growth, plant performance and cold tolerance under greenhouse. Phytotoxicity tests were carried out using higher concentrations (100, 1000 and 200 ppm) applied by dipping or spraying onto chili pepper plants. Deionized water controls were included. The results showed that the SBA materials did not affect seed germination; however, SBA-15 at 50 ppm and 100 ppm applied by imbibition significantly increased seedling height (up to 8-fold) and provided enhanced growth performance in comparison with controls under select treatment regimes. Weekly application of SBA-15 at 20 ppm significantly increased stem diameter and cold tolerance; however, SBA-16 showed significant decreases in plant height (20 ppm biweekly applied) and stem diameter (20, 50 and 100 ppm biweekly applied). The results demonstrate that both SBA materials provided hormetic effects in a dose dependent manner on chili pepper production and protection to cold stress. No phytotoxic response was evident. These findings suggested the nanostructured mesoporous silica have potential as a sustainable amendment strategy to increase crop production under stress-inducing cultivation conditions.