Beeswax Alcohol Prevents Low-Density Lipoprotein Oxidation and Demonstrates Antioxidant Activities in Zebrafish Embryos and Human Subjects: A Clinical Study.

Oxidative stress is one of the primary instigators of the onset of various human ailments, including cancers, cardiovascular diseases, and dementia. Particularly, oxidative stress severely affects low-density lipid & protein (LDL) oxidation, leading to several detrimental health effects. Therefore, in this study, the effect of beeswax alcohol (BWA) was evaluated in the prevention of LDL oxidation, enhancement of paraoxonase 1 (PON-1) activity of high-density lipid & protein (HDL), and zebrafish embryo survivability. Furthermore, the implication of BWA consumption on the oxidative plasma variables was assessed by a preliminary clinical study on middle-aged and older human subjects (n = 50). Results support BWA augmentation of PON-1 activity in a dose-dependent manner (10-30 µM), which was significantly better than the effect exerted by coenzyme Q10 (CoQ10). Moreover, BWA significantly curtails LDL/apo-B oxidation evoked by CuSO4 (final 0.5 µM) and a causes a marked reduction in lipid peroxidation in LDL. The transmission electron microscopy (TEM) analysis revealed a healing effect of BWA towards the restoration of LDL morphology and size impaired by the exposure of Cu2+ ions (final 0.5 µM). Additionally, BWA counters the toxicity induced by carboxymethyllysine (CML, 500 ng) and rescues zebrafish embryos from development deformities and apoptotic cell death. A completely randomized, double-blinded, placebo-controlled preliminary clinical study on middle- and older-aged human subjects (n = 50) showed that 12 weeks of BWA (100 mg/day) supplementation efficiently diminished serum malondialdehyde (MDA) and total hydroperoxides and enhanced total antioxidant status by 25%, 27%, and 22%, respectively, compared to the placebo-control and baseline values. Furthermore, the consumption of BWA did not exhibit any noteworthy changes in physical variables, lipid profile, glucose levels, and biomarkers pertinent to kidney and liver function, thus confirming the safety of BWA for consumption. Conclusively, in vitro, BWA prevents LDL oxidation, enhances PON-1 activity in HDL, and positively influences oxidative variables in human subjects.

Effect of mango butter on the physicochemical properties of beeswax-Moringa seed oil-based oleogels for topical application.

OBJECTIVE: The purpose of this research was to determine any connections between the characteristics of oleogels made of beeswax and the impact of mango butter. METHODS: Oleogel was prepared through inverted tube methods, and optimized through oil binding capacity. Other evaluations like bright field and polarized microscopy, Fourier-transform infrared (FTIR) spectroscopy, crystallization kinetics, mechanical study, and X-ray diffractometry (XRD). The drug release kinetic studies and in vitro antibacterial studies were performed. RESULTS: FTIR study reveals that the gelation process does not significantly alter the chemical composition of the individual components. Prepared gel exhibiting fluid-like behavior or composed of brittle networks is particularly vulnerable to disruptions in their network design. The incorporation of mango butter increases the drug permeation. In-vitro microbial efficacy study was found to be excellent. CONCLUSION: The studies revealed that mango butter can be used to modify the physico-chemical properties of the oleogels.

The Impact of Beeswax and Glycerol Monolaurate on Camellia Oil Oleogel's Formulation and Application in Food Products.

This study assessed the nutritional profile of camellia oil through its fatty acid composition, highlighting its high oleic acid content (81.4%), followed by linoleic (7.99%) and palmitic acids (7.74%), demonstrating its excellence as an edible oil source. The impact of beeswax (BW) and glycerol monolaurate (GML) on camellia oil oleogels was investigated, revealing that increasing BW or GML concentrations enhanced hardness and springiness, with 10% BW oleogel exhibiting the highest hardness and springiness. FTIR results suggested that the structure of the oleogels was formed by interactions between molecules without altering the chemical composition. In biscuits, 10% BW oleogel provided superior crispness, expansion ratio, texture, and taste, whereas GML imparted a distinct odor. In sausages, no significant differences were observed in color, water retention, and pH between the control and replacement groups; however, the BW group scored higher than the GML group in the sensory evaluation. The findings suggest that the BW oleogel is an effective fat substitute in biscuits and sausages, promoting the application of camellia oil in food products.

Impact of binary mixtures of natural waxes in mechanical properties and microstructure of oleogels.

BACKGROUND: Solid fats are critical to obtaining a wide range of food texture and quality characteristics, but their consumption is strongly associated with higher cardiovascular disease risks. Structuring unsaturated oils with natural waxes into oleogels (OG) is an innovative solution to develop fat mimics with a healthier profile. RESULTS: Soy wax (SW), beeswax (BW) and carnauba wax (CW), have been used in binary mixtures of waxes, aiming to understand their interactions and influence on OG quality properties and microstructural characteristics. In the present study, OGs were produced using binary wax mixtures and analyzed for texture, color, smoke point, microstructure, Fourier-transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD). Wax combinations led to antagonistic (mixtures with SW) and synergistic interactions (BW/CW) based on their mechanical properties. At the microstructural level BW/CW blends showed a reduction in crystal size and with a more compact structure. XRD and FTIR spectra revealed a packing of orthorhombic perpendicular subcell for most OGs, whereas SW produced samples with an arrangement with ß' crystals, characteristic of edible solid fats. Additionally, when compared to commercial beef fat, BW/CW mixtures showed similar quality attributes indicating that they could act as fat mimic. CONCLUSION: The combined analysis of microstructure, spectroscopic and mechanical properties enhanced the understanding of how the nature of the interactions between waxes and lipid phases impact in the final quality of the structured oils. The study's insights indicate that binary wax combinations can efficiently replace solid fats, offering healthier alternatives at the same time as preserving desired sensory characteristics. © 2024 Society of Chemical Industry.

Seasonal screening of pesticide residues in beehive products collected from different districts in Egypt.

Pesticides are of immense importance in agriculture, but they might contaminate bees' products. In this study, samples of honey, pollen, and beeswax were collected, seasonally, from apiaries in Toshka (Aswan), El-Noubariya (El-Beheira), and Ismailia (Ismailia) cities in Egypt. The pesticide residues were analyzed using the GC-MS after being extracted and cleaned using the QuEChERS method. Results showed that samples from El-Noubariya had great content of residues followed by Ismailia, and finally Toshka. Samples collected during fall and winter had the highest pesticide residue contents. Specifically, the phenylconazole fungicide group was repeatedly detected in all the examined samples along with organophosphate insecticides. Beeswax samples had the greatest amounts of pesticide residues followed by pollen and then honey samples. Chlorpyrifos (0.07-39.16 ng/g) and profenofos (1.94-17.00 ng/g) were detected in honey samples and their products. Pyriproxyfen (57.12 ng/g) and chlorpyrifos-methyl (39.16 ng/g) were detected in great amounts in beeswax samples from Ismailia and El-Noubariya, respectively. Yet, according to health hazard and quotient studies, the amounts of pesticides detected in honey do not pose any health threats to humans.

Antimicrobial activity and wound healing effect of a novel natural ointment: an in vitro and in vivo study.

Infection and pathological disorders, such as cellular disorders, ischaemia, neuropathy and angiogenesis, are considered the most critical factors which cause a delay in the wound healing process in patients with diabetes. This study aimed to investigate the effect of an ointment based on ostrich oil containing honey, beeswax, and ethanolic extracts of Nigella sativa, propolis and Cassia angustifolia on the wound healing process of diabetic rats. Gas chromatography/mass spectrometry analysis showed caffeic acid and pinostrobin chalcone molecules present in propolis, giving antibacterial and antifungal properties to the compound. The antibacterial assessment showed the ointment had remarkable antibacterial activity against Staphylococcus aureus (8.6±0.28mm), Escherichia coli (9.4±0.31mm), Acinetobacter baumannii (7.2±0.23mm) and Pseudomonas aeruginosa (13.9±0.42mm). In vivo results showed the ointment significantly accelerated wound healing and increased collagen deposition compared with the control (p<0.05). Histopathology evaluation also showed hair follicles, sebaceous glands and vessels in the group that used the ointment. These results proved successful and diabetic wound healing was rapid. Therefore, it could be concluded that the fabricated ointment could be a suitable candidate for wound healing.

Evaluation of high oleic sunflower oil oleogels with beeswax, beeswax-glyceryl monopalmitate, and beeswax-Span80 in cookie preparation.

BACKGROUND: Shortening is used widely in cookie preparation to improve quality and texture. However, large amounts of saturated and trans fatty acids present in shortening have adverse effects on human health, and much effort has been made to reduce the use of shortening. The use of oleogels might be a suitable alternative. In this study, the oleogels of high oleic sunflower oil with beeswax (BW), BW-glyceryl monopalmitate (BW-GMP), and BW-Span80 (BW-S80) were prepared and their suitability to replace shortening in cookie preparation was evaluated. RESULTS: The solid fat content of BW, BW-GMP, and BW-S80 oleogels was significantly lower than that of commercial shortening when the temperature was not higher than 35 °C. However, the oil-binding capacity of these oleogels was almost similar to that of shortening. The crystals in the shortening and oleogels were ß' form mainly; however, the morphology of crystal aggregates in these oleogels was different from that of shortening. The textural and rheological properties of doughs prepared with the oleogels were similar, and clearly different from those of dough with commercial shortening. The breaking strengths of cookies made with oleogels were lower than that of cookies prepared with shortening. However, cookies containing BW-GMP and BW-S80 oleogels were similar in density and color to those prepared with shortening. CONCLUSION: The textural properties and color of cookies with BW-GMP and BW-S80 oleogels were very similar to those of the cookies containing commercial shortening. The BW-GMP and BW-S80 oleogels could act as alternatives to shortening in the preparation of cookies. © 2023 Society of Chemical Industry.

Heavy Metal Concentrations of Beeswax (Apis mellifera L.) at Different Ages.

Beeswax is a naturally occurring product that worker bees produce. Beeswax is used in a variety of industries and pharmaceuticals. Humans utilize it extensively in cosmetics, medicinal formulations, and food manufacturing. Beeswax is an essential component of advanced contemporary beekeeping. Beekeepers, in particular, utilize significant amounts of beeswax to make beeswax comb foundation. In its natural condition, beeswax is white, but it becomes yellow then dark in color when it comes into touch with honey and pollen. The ongoing use of wax comb in bee activities (such as brood rearing, storage honey and bee bread), combined with environmental factors such as heavy metal and pesticide residues, resulted in a black color. Because of heavy metals can accumulate in wax for decades, beeswax can be a helpful tool for gathering data on hazardous contaminants in the environment. Because of their lipid-based chemical composition, beeswax combs act as a sink for numerous ambient pollutants as well as poisons when in the hive. The current study aims to measure nine heavy metals and important elements, including iron (Fe), chromium (Cr), zinc (Zn), copper (Cu), nickel (Ni), manganese (Mn), lead (Pb), cadmium (Cd), and cobalt (Co) in beeswax collected in the Behaira governorate region of Egypt between 2018 and 2022. Sample collection was conducted each year in triplicate. The samples were analyzed using an atomic absorption spectrophotometer. The quantity of metals in beeswax at different ages differed significantly. Depending on the wax age, Fe has the highest concentration in the range of 2.068 to 5.041 ppm, while Cd has the lowest ratio at 0.024 to 0.054 ppm from the first to fifth years old of comb age. The findings showed that as beeswax combs aged, the concentration of heavy metals rose. According to the study, it should gradually recycle beeswax combs each year and also adding new foundations.

Quantitation of 1,4-Dichlorobenzene and Thymol in Beeswax Using Dynamic Headspace Vacuum Transfer in Trap Extraction Prior to Gas Chromatography-Mass Spectrometry.

A method based on dynamic headspace vacuum transfer in trap extraction, followed by gas chromatography coupled with a mass spectrometer (DHS-VTT-GC-MS), was validated for the fast quantitation of 1,4-dichlorobenzene (p-dichlorobenzene; PDCB) and thymol residues in beeswax. The quantitation limits (LOQ) were 0.05 mg/kg (PDCB) and 0.25 mg/kg (thymol). Recoveries above 80% were obtained for PDCB concentrations between 0.05 and 10 mg/kg and for thymol concentrations between 0.25 and 200 mg/kg. Analysis of beeswax samples showed a good correlation between the results obtained by DHS-VTT-GC-MS analysis and those of a previous method based on SPE extraction followed by gas chromatography and triple- quadrupole mass spectrometry (GC-MS/MS) (R2 = 0.9770 for PDCB and 0.9666 for thymol). However, the sample preparation and chromatography were much shorter using the DHS-VTT-GC-MS method. Forty comb foundations samples produced in Switzerland in 2019 and 2021 were analysed using DHS-VTT-GC-MS. Fourteen samples contained PDCB above the LOQ of 0.05 mg/kg, ranging up to a maximum of 1.53 mg/kg. The mean concentration of the positive samples was 0.22 mg/kg. All samples contained thymol residues ranging between 3.9 and 84.4 mg/kg with mean and median concentrations of 22.7 mg/kg and 17.4 mg/kg. Residue levels of PCDB and thymol in Swiss beeswax were substantially below those measured 20 (PDCB) and 10 (thymol) years ago, respectively.

[Formation of the organoleptic profile of sugar cookies with a modified fat component].

Currently, as part of solving the problems of improving the quality and safety of food, research is underway on the introduction of oleogels into foodstuffs to replace solid fats, which contain saturated and trans-isomeric fatty acids. However, such a replacement leads to a change in the organoleptic characteristics of food. The aim of the work was to identify the characteristics that affect the difference in the perception of cookie descriptors baked using hard fat and oleogels. Material and methods. Three batches of sugar cookies samples with different fat components were produced for the study: butter cookies; cookies based on oleogel structured with beeswax; and cookies based on oleogel structured with combinations of beeswax fractions. Organoleptic evaluation was carried out by two methods: the triangle method according to ISO 4120:2004 and the free-choice profiling method according to ISO 13299-2015. In this case, the profile was derived statistically, by means of a generalized Procrustes analysis. Results and discussion. The analyze of cookies samples using the triangle method showed there was no noticeable difference between the control cookies samples and cookies with oleogel structured with beeswax, but there were significant differences with cookies baked with the use of oleogel structured with a combination of fractions. Generalized Procrustes analysis was used to interpret the results of the descriptor profile analysis. It was shown that differences in perception between samples were due to the severity of the following descriptors: Creamy flavour, Fat flavour, Floral flavour, Waxy flavour, Shape. A close correlation has been established between individual descriptors characterizing the samples under study. Conclusion. The data obtained as a result of the conducted studies allow not only to judge the acceptability of the use of oleogels in cookies from an organoleptic point of view, but also to identify individual characteristics that affect the perception of cookie samples. In the future, this information can be used to optimize the formulation of finished cookies containing oleogels in order to change the organoleptic profile in a targeted manner.

Analysis of balms taken from Egyptian human mummies using solid-phase extraction and gas chromatography-mass spectrometry.

The aim of this paper is to establish a protocol by solid-phase extraction-gas chromatography-mass spectrometry leading to a wide and fine qualitative chemical characterization of the several natural substances present in human mummies' balms, using a minimal quantity of samples. In this study, nine samples were analyzed from mummies dating back from the Third Intermediate Period to the Roman Period, and were provided by the Confluences Museum (Lyon, France). Using solid-phase extraction, three fractions were examined in this protocol. The first one, eluted with hexane, concerned chemical families of hydrocarbons of bitumen. The second, eluted with ethanol, enabled terpenic compounds to be characterized and beeswax. The last one, composed of diethyl ether with 2% acetic acid, extracted carboxylic acids with a long aliphatic chain (fatty matter) and glycerides. This study also allowed the characterization of non-saponified compounds from beeswax to be obtained while excluding the common saponification step. The analyzed mummification balms were shown to contain fatty matter, beeswax, bitumen, and diterpenic resinous material. This one-pot solid-phase extraction-gas chromatography-mass spectrometry method was efficient in reducing both the number of analytical steps and the complexity of the archaeological balms subsequently analyzed by gas chromatography-mass spectrometry.

Bee products as a source of promising therapeutic and chemoprophylaxis strategies against COVID-19 (SARS-CoV-2).

The emergence of novel coronavirus (SARS-CoV-2) in 2019 in China marked the third outbreak of a highly pathogenic coronavirus infecting humans. The novel coronavirus disease (COVID-19) spread worldwide, becoming an emergency of major international concern. However, even after a decade of coronavirus research, there are still no licensed vaccines or therapeutic agents to treat the coronavirus infection. In this context, apitherapy presents as a promising source of pharmacological and nutraceutical agents for the treatment and/or prophylaxis of COVID-19. For instance, several honeybee products, such as honey, pollen, propolis, royal jelly, beeswax, and bee venom, have shown potent antiviral activity against pathogens that cause severe respiratory syndromes, including those caused by human coronaviruses. In addition, the benefits of these natural products to the immune system are remarkable, and many of them are involved in the induction of antibody production, maturation of immune cells, and stimulation of the innate and adaptive immune responses. Thus, in the absence of specific antivirals against SARS-CoV-2, apitherapy could offer one hope toward mitigating some of the risks associated with COVID-19.

An Insight to the Composition of Pre-Hispanic Mayan Funerary Pigments by 1H-NMR Analysis.

The funerary rites of particular members of the pre-Hispanic Mayan society included the pigmentation of the corpse with a red color. In order to understand this ritual, it is first necessary to identify the constituents of the pigment mixture and then, based on its properties, analyze the possible form and moment of application. In the present approach, 1H-NMR analysis was carried to detect organic components in the funerary pigments of Xcambó, a small Maya settlement in the Yucatan Peninsula. The comparison of the spectra belonging to the pigment found in the bone remains of seven individuals, and those from natural materials, led to the identification of beeswax and an abietane resin as constituents of the pigment, thus conferring it agglutinant and aromatic properties, respectively. The 1H-NMR analysis also allowed to rule out the presence of copal, a resin found in the pigment cover from paramount chiefs from the Mayan society. Additionally, a protocol for the extraction of the organic fraction from the bone segment without visible signs of analysis was developed, thus broadening the techniques available to investigate these valuable samples.

Lyophilized Drug-Loaded Solid Lipid Nanoparticles Formulated with Beeswax and Theobroma Oil.

Solid lipid nanoparticles (SLNs) have the potential to enhance the systemic availability of an active pharmaceutical ingredient (API) or reduce its toxicity through uptake of the SLNs from the gastrointestinal tract or controlled release of the API, respectively. In both aspects, the responses of the lipid matrix to external challenges is crucial. Here, we evaluate the effects of lyophilization on key responses of 1:1 beeswax-theobroma oil matrix SLNs using three model drugs: amphotericin B (AMB), paracetamol (PAR), and sulfasalazine (SSZ). Fresh SLNs were stable with sizes ranging between 206.5-236.9 nm. Lyophilization and storage for 24 months (4-8 °C) caused a 1.6- and 1.5-fold increase in size, respectively, in all three SLNs. Zeta potential was >60 mV in fresh, stored, and lyophilized SLNs, indicating good colloidal stability. Drug release was not significantly affected by lyophilization up to 8 h. Drug release percentages at end time were 11.8 ± 0.4, 65.9 ± 0.04, and 31.4 ± 1.95% from fresh AMB-SLNs, PAR-SLNs, and SSZ-SLNs, respectively, and 11.4 ± 0.4, 76.04 ± 0.21, and 31.6 ± 0.33% from lyophilized SLNs, respectively. Thus, rate of release is dependent on API solubility (AMB < SSZ < PAR). Drug release from each matrix followed the Higuchi model and was not affected by lyophilization. The above SLNs show potential for use in delivering hydrophilic and lipophilic drugs.

Evaluation of beeswax coated coconut shells for packaging of set-type dahi: an Indian fermented product.

An attempt was made to develop coconut shell-based eco-friendly packaging material for the packaging of set-type dahi. Beeswax was used to improve the moisture barrier properties of coconut shell (CS). UV treatment for 60 min showed complete disinfection of the coconut shell surface. The top open side of the coconut shell was sealed with aluminium foil using pectin solution as adhesive. Set-type dahi was prepared and packaged in beeswax coated CS (BCCS) and evaluated for physico-chemical, textural, sensorial and microbiological quality changes for a period of 21 days at 6 ± 1 °C. The rate of increase in acidity was found slower in BCCS as compared to dahi packaged in polypropylene cups (control). There was no whey separation, consequently water-holding capacity and firmness of product were better in dahi packaged in BCCS. The sensorial analysis revealed better body and texture properties for dahi packaged in BCCS. However, the flavour score of dahi packaged in BCCS decreased comparatively after 12 days due to the absorption of coconut flavor by the product. These findings demonstrated the potential of coconut shell based eco-friendly packaging material for packaging set-type dahi and thereby aim to reduce the use of non-biodegradable plastics.

Protocol for a systematic review and meta-analysis of human exposure to pesticide residues in honey and other bees' products.

BACKGROUND: The presence of pesticides in honey and related products is an increasing concern for consumers and producers, although there is lack of data on the current burden of exposure of the general human population through these products. We present a protocol for a systematic review and meta-analysis of contamination to insecticides, herbicides and fungicides of products from honeybees, and an estimation of how much the consumption of these products contributes to the ADI (Acceptable Daily Intake) of selected substances. OBJECTIVES: We aim to systematically review and meta-analyse studies on the contamination to plant protection products in honey, royal jelly, beeswax and propolis, applying the Navigation Guide and WHO-ILO systematic review methodology as an organizing framework. DATA SOURCES: We will search electronic academic databases for potentially relevant records from PubMed, TOXNET and EMBASE. We will include quantitative studies analysing the contamination from insecticides, herbicides and fungicides in honey, propolis, royal jelly and beeswax. In particular, we will evaluate the presence of the following substances and classes of pesticides: Glyphosate, Chlorpyrifos, pyrethroid and neonicotinoid pesticides, fungicides and acaricides. STUDY APPRAISAL AND SYNTHESIS METHODS: At least two authors will independently screen titles and abstracts at a first stage of review, and full texts at a second stage, of potentially eligible records against the eligibility criteria; data extraction of included studies will then be performed by at least two authors, in blind. At least two authors will assess risk of bias and the quality of evidence, using the most suited tools currently available. The data on prevalence of contaminated samples and concentration of pesticides in the products will be combined using meta-analysis: when more than three studies reporting the necessary measures to fit the models are available, meta-analysis will be performed separately by product and by exposure; otherwise, weighted descriptive analysis will be performed. We will report the results using the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines (PRISMA).

Evaluation of linseed oil oleogels to partially replace pork backfat in fermented sausages.

BACKGROUND: Nowadays, fat replacement in meat products is a matter of concern in the meat industry. The objective of this study was to evaluate the replacement of pork backfat with two oleogels of linseed in dry-cured sausages. RESULTS: Five batches of dry-cured sausages were prepared with two oleogels, a mixture of γ-oryzanol and ß-sitosterol (SO) and beeswax (B), at two levels of replacement (20% and 40%) (SO-20, SO-40, B-20, and B-40, respectively) and a control batch. The fatty acid profile improved in terms of nutrition: the polyunsaturated fatty acid / saturated fatty acid (PUFA/SFA) and n-6/n-3 ratio was about 1.41 and 0.93 for the higher levels of replacement, SO-40 and B-40, respectively. Quality parameters such as pH and color also changed with the inclusion of oleogels, resulting in changes in the sensory quality. CONCLUSION: Oleogels based on linseed enabled the replacement of pork backfat in fermented sausages. Depending on the level of fat substitution, such oleogels could replace fat in dry-cured sausages at the industrial level. © 2019 Society of Chemical Industry.

Release-controlled microcapsules of thiamethoxam encapsulated in beeswax and their application in field.

Using beeswax as wrapping matrix, two types of release-controlled TM (thiamethoxam)/BK(beeswax-kaolin) microcapsules were prepared by adsorbing TM on kaolin and then encapsulated with beeswax, or directly wrapping TM with beeswax. The structure and morphology of the TM/BK microcapsules were characterized. The effects of different preparation methods, the particle size, pH conditions and different additives on the release property of the TM/BK microcapsules were investigated in water and soil column to compare the advantages of the two approaches. Finally, the insecticidal effect of the TM/BK microcapsules against sugarcane borer and rice planthopper was tested. The results show that the TM/BK microcapsules have a better sustained-release in both water and soil, and the release rate is different under different pH conditions. In addition, the releasing time of the TM/BK microcapsules can be modified by different preparation methods and combination of different additives. In the field applications, the insecticidal activity of the TM/BK microcapsules was better than that of non-sustained control group. Especially in the rice field test, 45 days after the application, the control group lost the activity against rice planthopper because of drug loss, whereas the TM/BK microcapsule group still retained about 90% of the insecticidal activity. The results suggest that the microcapsules have better agricultural application for insect control.

Embryonic stem cells differentiated into neuron-like cells using SB431542 small molecule on nanofibrous PLA/CS/Wax scaffold.

Electrospun nanofibrous scaffolds show huge potential to improve the neurological outcome in central nervous system disorders. In this study, we cultured mouse embryonic stem cells (mESCs) on an electrospun nanofibrous polylactic acid/Chitosan/Wax (PLA/CS/Wax) scaffold and surveyed the attachment, behavior, and differentiation of mESCs into neural cells. Differentiation in neural-like cells (NLCs) was investigated with a medium containing SB431542 as a small molecule and conjugated linolenic acid after 20 days. We used Immunocytochemistry and quantitative real-time polymerase chain reaction (RT-PCR) techniques to assess neural marker expression in differentiated cells. SEM imaging demonstrated that mESCs could strongly attach, stretch, and differentiate on PLA/CS/Wax scaffolds. MESCs that were cultured on PLA/CS/Wax scaffolds showed enhanced numbers of neural structures and neural markers including Nestin, NF-H, Tuj-1, and Map2 in neural induction medium compared to the control sample. These results revealed that electrospun PLA/CS/Wax scaffolds associated with the induction medium can assemble proper conditions for stem cell differentiation into NLCs. We hope that the development of new technologies in neural tissue engineering may pave a new avenue for neural tissue regeneration.