Assessment of acute toxicity of crude extract rich in carotenoids from Cantaloupe melon (Cucumis melo L.) and the gelatin-based nanoparticles using the zebrafish (Danio rerio) model.

Cantaloupe melon is known for its carotenoid-rich orange pulp. However, carotenoids are sensitive to oxygen, light, and heat, potentially reducing their benefits. Nanoencapsulation can preserve these benefits but raises concerns about toxicity. We aimed to assess the safety and bioactive potential of crude extract-rich carotenoids (CE) and nanoparticles based on gelatin loaded with CE (EPG) by investigating parameters such as cardio or neurotoxicity, especially acute toxicity. EPG was obtained by O/W emulsification and characterized by different methods. Zebrafish embryos were exposed to CE and EPG at 12.5 mg/L and 50 mg/L for 96h and were investigated for survival, hatching, malformations, and seven days post fertilization (dpf) larvae's visual motor response. Adult fish underwent behavioral tests after acute exposure of 96h. CE and EPG showed no acute toxicity in zebrafish embryos, and both improved the visual motor response in 7dpf larvae (p = 0.01), suggesting the potential antioxidant and provitamin A effect of carotenoids in cognitive function and response in the evaluated model. Adult fish behavior remained with no signs of anxiety, stress, swimming pattern changes, or sociability that would indicate toxicity. This study highlights the safety and potential benefits of carotenoids in zebrafish. Further research is needed to explore underlying mechanisms and long-term effects.

Nanoparticles Loaded with a Carotenoid-Rich Extract from Cantaloupe Melon Improved Hepatic Retinol Levels in a Diet-Induced Obesity Preclinical Model.

The study evaluated the effect of the carotenoid-rich extract from cantaloupe melon (CE) nanoencapsulated in porcine gelatin (EPG) on hepatic retinol concentration and liver damage scores in Wistar rats with obesity induced by high glycemic index and high glycemic load diet (HGLI diet). For 17 days, animals were fed the HGLI diet. They were divided into three groups and treated for 10 days [HGLI diet + water, HGLI diet + CE (12.5 mg/kg), and HGLI diet + EPG (50 mg/kg)]. The groups were evaluated for dietary intake, retinol, weight variation, hematological parameters, fasting glucose, lipid profile, hepatic retinol concentration, AST/ALT ratio, FIB-4 (Fibrosis-4 Index for Liver Fibrosis), and APRI (AST to Platelet Ratio Index) scores to evaluate the effects on the liver. Animals treated with EPG showed a lower dietary intake (p < 0.05). No significant weight change was detected in the evaluated groups (p > 0.05). The EPG-treated group had significantly higher concentrations (p < 0.05) of hepatic retinol [266 (45) µg/g] than the untreated group [186 (23.8) µg/g] and the one treated with CE [175 (8.08) µg/g]. Liver damage assessment scores did not show significant differences, but the lowest means were observed in the group treated with EPG. The nanoencapsulation of the extract rich in beta-carotene promoted reduced food consumption and increased hepatic retinol without causing significant changes in liver damage scores. Thus, EPG is a candidate for future clinical studies to evaluate the beneficial effects of treating diseases involving vitamin A deficiencies.

Efficacy of Carotenoid-Loaded Gelatin Nanoparticles in Reducing Plasma Cytokines and Adipocyte Hypertrophy in Wistar Rats.

The present study investigated the effect of gelatin-based nanoparticles (EPG) loaded with a carotenoid-rich crude extract (CE) on systemic and adipose tissue inflammatory response in a model with inflammation induced by a high glycemic index and high glycemic load diet (HGLI). Nanoparticles synthesized were characterized by different physical and chemical methods. The in vivo investigation evaluated Wistar rats (n = 20, 11 days, adult male with 21 weeks) subdivided into untreated (HGLI diet), conventional treatment (nutritionally adequate diet), treatment 1 (HGLI + crude extract (12.5 mg/kg)), and treatment 2 (HGLI + EPG (50 mg/kg)) groups. Dietary intake, caloric intake and efficiency, weight, inflammatory cytokines tissue concentration, visceral adipose tissue (VAT) weight, histopathological analysis, and antioxidant activity in plasma and VAT were investigated. EPG showed the same physical and chemical characteristics as previous batches (95.2 nm, smooth surface, and chemical interactions between materials). The EPG-treated group was the only group promoting negative ∆dietary intake, ∆caloric efficiency, and ∆weight. In addition, it presented a significant reduction (p < 0.05) in IL-6 and leptin levels and a greater presence of multilocular adipocytes. The results suggest that EPG can act as a nutraceutical in adjuvant therapy for treating inflammatory diseases associated with adipose tissue accumulation.

In silico structure-based design of peptides or proteins as therapeutic tools for obesity or diabetes mellitus: A protocol for systematic review and meta analysis.

BACKGROUND: In silico studies using dynamic simulation or molecular docking have boosted the screening and identification of molecules and/or targets in studies aimed at treating diseases such as obesity and diabetes mellitus, optimizing the development of new drugs. This study aims to describe a systematic review protocol on peptides and proteins evaluated in silico as potential therapeutic agents for obesity or diabetes mellitus. METHODS: This protocol followed the Preferred Reporting Items for Systematic Reviews and Meta-analyses Protocols and was registered in the International Prospective Register of Systematic Reviews database (number: CRD42022355540). The databases to be searched will be PubMed, ScienceDirect, Scopus, Web of Science, virtual health library, and EMBASE. It will be included in silico studies that evaluate the simulation by dynamics or molecular docking of proteins or peptides involved in treating obesity or diabetes mellitus. Two independent reviewers will select studies, extract data, and assess methodological quality using the adapted Strengthening the reporting of empirical simulation studies. A narrative synthesis of the included studies will be performed for the systematic reviews. RESULTS: This protocol contemplates the production of 2 systematic reviews to be developed focusing on obesity or diabetes mellitus. CONCLUSION: The reviews will enable knowledge of peptides and proteins involved in research treating these diseases and will emphasize the importance of in silico studies in this context and for the development of future studies.

In silico structure-based designers of therapeutic targets for diabetes mellitus or obesity: A protocol for systematic review.

Obesity is a significant risk factor for several chronic non-communicable diseases, being closely related to Diabetes Mellitus. Computer modeling techniques favor the understanding of interaction mechanisms between specific targets and substances of interest, optimizing drug development. In this article, the protocol of two protocols of systematic reviews are described for identifying therapeutic targets and models for treating obesity or diabetes mellitus investigated in silico. The protocol is by the guidelines from the Preferred Reporting Items for Systematic Reviews and Meta-Analyzes Protocols (PRISMA-P) and was published in the International Prospective Register of Systematic Reviews database (PROSPERO: CRD42022353808). Search strategies will be developed based on the combination of descriptors and executed in the following databases: PubMed; ScienceDirect; Scopus; Web of Science; Virtual Health Library; EMBASE. Only original in silico studies with molecular dynamics, molecular docking, or both will be inserted. Two trained researchers will independently select the articles, extract the data, and assess the risk of bias. The quality will be assessed through an adapted version of the Strengthening the Reporting of Empirical Simulation Studies (STRESS) and the risk of bias using a checklist obtained from separate literature sources. The implementation of this protocol will result in the elaboration of two systematic reviews identifying the therapeutic targets for treating obesity (review 1) or diabetes mellitus (review 2) used in computer simulation studies and their models. The systematization of knowledge about these treatment targets and their in silico structures is fundamental, primarily because computer simulation contributes to more accurate planning of future either in vitro or in vivo studies. Therefore, the reviews developed from this protocol will guide decision-making regarding the choice of targets/models in future research focused on therapeutics of obesity or Diabetes Mellitus contributing to mitigate of factors such as costs, time, and necessity of in vitro and/or in vivo assays.

Nanoparticles Containing Tamarind Isolate Protein Potentiate the Satiety without Promoting the Anti-Inflammatory Effect in a Preclinical Model of Diet-Induced Obesity.

The study aimed to evaluate the nanoparticles (ECW) containing tamarind trypsin inhibitor (TTI) concerning the storage effect under different conditions on antitrypsin activity and the bioactive potential in a preclinical model. ECW was exposed to different pH and temperatures to evaluate the interaction between TTI and its encapsulating agents, monitored by antitrypsin activity. Wistar rats (n = 25) with obesity induced by diet were divided into groups: untreated; treatment with nutritionally adequate diet; treatment with nutritionally adequate diet and ECW/12.5 mg/kg; treatment with ECW/12.5 mg/kg; and treatment with TTI/25 mg/kg. The groups were evaluated over ten days with regards to satiety, zoometric, biochemical, and inflammatory parameters, using ten times less TTI (2.5 mg/kg) contained in ECW. TTI was protected and encapsulated in ECW without showing residual inhibitory activity. Only at gastric pH did ECW show antitrypsin activity. At different temperatures, it showed high antitrypsin activity, similar to TTI. The animals treated with ECW had significantly reduced body weight variation (p < 0.05), and only TTI treatment reduced the inflammatory parameters significantly (p < 0.05). The study showed that by using lower concentrations of TTI in ECW it was possible to perceive promising effects with perspectives of use in functional products for managing obesity and its complications.

Encapsulation techniques perfect the antioxidant action of carotenoids: A systematic review of how this effect is promoted.

Considering that antioxidant activities are directly related to carotenoid functionalities, it is necessary to use techniques that promote the stability of these natural pigments. This systematic review aimed to gather evidence on the effect of encapsulation techniques on the maintenance and/or enhancement of the antioxidant activity of carotenoids. The study was registered in PROSPERO (CRD42020142065). Searches were performed in PubMed, Embase, Virtual Health Library, Scopus, ScienceDirect, and Web of Science databases. Assessment of methodological quality was performed using OHAT. A total of 1577 articles were selected, resulting in 20 eligible studies. Overall, results showed that the mechanisms involved are related to the emergence of new chemical interactions, increased surface area, and the controlled release of carotenoids. Thus, evidence proved that encapsulation could preserve and/or enhance bioactivities of carotenoids, allowing the use in foods to promote benefits on population health.

Antioxidant stability enhancement of carotenoid rich-extract from Cantaloupe melon (Cucumis melo L.) nanoencapsulated in gelatin under different storage conditions.

The study evaluated the potential and antioxidant stability of nanoencapsulated carotenoid-rich extract (CE) from Cantaloupe melon (EPG). DPPH and ABTS radical scavenging assays were used to investigate the nanoencapsulation effect on antioxidant potential. CE and EPG stability were evaluated at 25 °C and 5 °C, with and without light (1600 lx) for 60 days, determining the ß-carotene concentration by UHPLC and antioxidant potential by ABTS. The antioxidant potential of carotenoids increased after nanoencapsulation (57-59%). After 60 days, there was low retention of ß-carotene (0-43.6%) in the CE, mainly at 25 °C light (0.00%) and dark (10.0%), and total loss of activity in the four conditions. EPG preserved the ß-carotene concentration in the dark at 25 °C (99.0%) and in the light (83.1%) and dark (99.0%) at 5 °C, maintaining the antioxidant potential (68.7-48.3%). Therefore, EPG enhanced and stabilized the antioxidant potential of carotenoids, beneficial to human health.

Safety and bioactive potential of nanoparticles containing Cantaloupe melon (Cucumis melo L.) carotenoids in an experimental model of chronic inflammation.

The safety and bioactive potential of crude carotenoid extract from Cantaloupe melon nanoencapsulated in porcine gelatin (EPG) were evaluated in a chronic inflammatory experimental model. Animals were fed a high glycemic index and high glycemic load (HGLI) diet for 17 weeks and treated for ten days with 1) HGLI diet, 2) standard diet, 3) HGLI diet + crude carotenoid extract (CE) (12.5 mg/kg), and 4) HGLI diet + EPG (50 mg/kg). General toxicity signals were investigated, considering body weight, food intake, hematological, biochemical parameters, relative weight, morphology, and histopathology of organs. The biochemical parameters indicated the low toxicity of EPG. Acute hepatitis was observed in animals' livers, but CE and EPG groups presented improved tissue appearance. Chronic enteritis was observed in animals, with villi and intestinal glands preservation in the EPG group. The results suggest the safety and the bioactive effect of EPG, possibly related to its anti-inflammatory potential.

Gelatin nanoparticles enable water dispersibility and potentialize the antimicrobial activity of Buriti (Mauritia flexuosa) oil.

BACKGROUND: Buriti oil presents numerous health benefits, but due to its lipophilic nature and high oxidation, it is impossible to incorporate it into aqueous food matrices. Thus, the present study evaluated whether powder nanoparticles based on porcine gelatin (OPG) and in combination with sodium alginate (OAG) containing buriti oil obtained by O/W emulsification followed by freeze-drying enabled water dispersibility and preserved or increased the antimicrobial activity of the oil. RESULTS: OPG presented spherical shape, smooth surface, smaller particle size and polydispersity index [51.0 (6.07) nm and 0.40 (0.05)], and better chemical interaction between the nonpolar amino acids and the hydrophobic oil chain. OPG also presented a higher dispersibility percentage [85.62% (7.82)] than OAG [50.19% (7.24)] (p < 0.05), and significantly increased the antimicrobial activity of the oil by 59, 62, and 43% for Pseudomonas aeruginosa, Klebsiella pneumonia, and Staphylococcus aureus, respectively. CONCLUSIONS: Thus, nanoencapsulation in gelatin is a promising strategy to increase the potential to use buriti oil in foods.

Effect of carotenoid encapsulation on antioxidant activities: A protocol for systematic review.

BACKGROUND: Carotenoids play essential roles in human health, such as antioxidant activity, and therefore can decrease free radicals oxidation action, preventing numerous diseases. However, these compounds have an unstable nature, turning them susceptible to adverse conditions in food processing and storage. Thereby the search for alternatives that maintain and enhance carotenoid antioxidant function, such as encapsulation, has grown. The objective of this study was to establish a systematic review protocol to evaluate the effect of different encapsulation techniques on the antioxidant action of carotenoids, evaluating which one is the best and safest, and their role in enhancing the antioxidant activity. METHODS: This protocol was guided by the preferred reporting items for protocols for systematic reviews and meta-analyzes. The databases to be searched are PubMed, EMBASE, Scopus, ScienceDirect, and Web of Science. Experimental studies conducted in rats and mice (in vivo) of both sexes and ages, evaluating the use of encapsulated and crude carotenoids will be included in the systematic review. The characteristics of the studies, the experimental model, and the main results will be described, and the risk of bias assessment will be evaluated. Three independent reviewers will proceed with the selection of studies, data extraction, and methodological quality assessment. A narrative synthesis will be made for the included studies. Besides, if sufficient qualitative data is available, a meta-analysis will be conducted. I2 statistics will be used to assess heterogeneity. RESULTS: This protocol will guide the production of a systematic review that can determine the effect of different encapsulation techniques and encapsulating agents on the antioxidant action of carotenoids. Thus, it will enable the determination of the best encapsulation techniques to promote the preservation and increase of the antioxidant activity, contributing to future research that may reproduce the best carotenoid encapsulation technique in an animal model. CONCLUSION: The systematic review to be produced from this protocol will provide support for the construction of research that evaluates the effect of encapsulation on the antioxidant function of carotenoids and its possible application as a nutraceutical, considering that this functionality is directly associated with health promotion. RECORD OF SYSTEMATIC REVIEW: This review was recorded in the International Register of Prospective Systematic Reviews on January 22, 2020 (registration: CRD42020142065). Available at: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42020142065.

Nanoencapsulation improved water solubility and color stability of carotenoids extracted from Cantaloupe melon (Cucumis melo L.).

Cantaloupe melon carotenoids were encapsulated in porcine gelatin, whey protein isolate and concentrate by emulsification O/W to evaluate which agent could promote an increase in water solubility, and color stability in yogurt. The average particle size obtained was 59.3 (2.60) nm-161.0 (27.30) nm. Encapsulated crude extract in porcine gelatin presented the smallest size and polydispersity index [0.4 (0.04)], and showed sphericity, smooth surface and low agglomeration in SEM. These results associated to the good chemical interaction between the raw materials shown by FTIR, justify the increase in water solubility [0.072 (0.007) mg.mL-1] compared to the crude extract [0.026 (0.003) mg.mL-1]. The yogurt added with this nanoencapsulate remained stable for 60 days, unlike the crude extract. The results show that the nanoencapsulation using gelatin increased water solubility and the potential of application of melon carotenoids in food as natural dyes.