From bytes to bites: Advancing the food industry with three-dimensional food printing.

The rapid advancement of three-dimensional (3D) printing (i.e., a type of additive manufacturing) technology has brought about significant advances in various industries, including the food industry. Among its many potential benefits, 3D food printing offers a promising solution to deliver products meeting the unique nutritional needs of diverse populations while also promoting sustainability within the food system. However, this is an emerging field, and there are several aspects to consider when planning for use of 3D food printing for large-scale food production. This comprehensive review explores the importance of food safety when using 3D printing to produce food products, including pathogens of concern, machine hygiene, and cleanability, as well as the role of macronutrients and storage conditions in microbial risks. Furthermore, postprocessing factors such as packaging, transportation, and dispensing of 3D-printed foods are discussed. Finally, this review delves into barriers of implementation of 3D food printers and presents both the limitations and opportunities of 3D food printing technology.

Designing future foods: Harnessing 3D food printing technology to encapsulate bioactive compounds.

Bioactive compounds (BCs) provide numerous health benefits by interacting with one or more components of living tissues and systems. However, despite their potential health benefits, most of the BCs have low bioaccessibility and bioavailability, hindering their potential health-promoting activities. The conventional encapsulation techniques are time-consuming and have major limitations in their food applications, including the use of non-food grade chemicals, undesired sensory attributes, and storage stability issues. A cutting-edge, new technique based on 3D printing can assist in resolving the problems associated with conventional encapsulation technologies. 3D food printing can help protect BCs by incorporating them precisely into three-dimensional matrices, which can provide (i) protection during storage, (ii) enhanced bioavailability, and (iii) effective delivery and controlled release of BCs. Recently, various 3D printing techniques and inks have been investigated in order to create delivery systems with different compositions and geometries, as well as diverse release patterns. This review emphasizes the advances in 3D printing-based encapsulation approaches, leading to enhanced delivery systems and customized food formulations.

Conventional techniques for encapsulating bioactive compounds (BCs) are summarized.Potential applications of 3D food printing for encapsulating BCs are discussed.Extrusion-based 3D food printing can enhance the stability/bioavailability of BCs.Hydrogels are promising materials for 3D food printing-based encapsulation.Future prospects for integrating 3D printing with encapsulation are highlighted.

Polysaccharide-based porous biopolymers for enhanced bioaccessibility and bioavailability of bioactive food compounds: Challenges, advances, and opportunities.

Bioactive food compounds, such as lycopene, curcumin, phytosterols, and resveratrol, have received great attention due to their potential health benefits. However, these bioactive compounds (BCs) have poor chemical stability during processing and low bioavailability after consumption. Several delivery systems have been proposed for enhancing their stability and bioavailability. Among these methods, porous biopolymers have emerged as alternative encapsulation materials, as they have superior properties like high surface area, porosity, and tunable surface chemistry to entrap BCs. This reduces the crystallinity (especially for the lipophilic ones) and particle size, and in turn, increases solubilization and bioavailability. Also, loading BCs into the porous matrix can protect them against environmental stresses such as light, heat, oxygen, and pH. This review introduces polysaccharide-based porous biopolymers for improving the bioaccessibility/bioavailability of bioactive food compounds and discusses their recent applications in the food industry. First, bioaccessibility and bioavailability are described with a special emphasis on the factors affecting them. Then, porous biopolymer fabrication methods, including supercritical carbon dioxide (SC-CO2 ) drying, freeze-drying, and electrospinning and electrospraying, are thoroughly discussed. Finally, common polysaccharide-based biopolymers (i.e., starch, nanocellulose, alginate, and pectin) used for generating porous materials are reviewed, and their current and potential future food applications are critically discussed.

Production of high-purity phospholipid concentrate from buttermilk powder using ethanol-modified supercritical carbon dioxide.

A new strategy to concentrate phospholipids from buttermilk powder was developed using a food-grade green method based on ethanol-modified supercritical carbon dioxide (SC-CO2) extraction. The effects of extraction conditions, namely temperature (50 and 60°C), pressure (30 and 40 MPa), and ethanol concentration (10, 15, and 20%, wt/wt), on the total lipid yield and phospholipid content were investigated. The ethanol concentration had a more significant effect on the total lipid yield and phospholipid content than the temperature and pressure within the ranges studied. The highest phospholipid recovery was achieved at 60°C, 30 MPa, and 15% (wt/wt) ethanol with a total lipid yield of 6.3% (wt/wt), of which 49% (wt/wt) were phospholipids composed of dihydrosphingomyelin (5%), sphingomyelin (24%), phosphatidylethanolamine (22%), phosphatidylserine (2%), phosphatidylinositol (3%), and phosphatidylcholine (44%). The triacylglycerol compositions of extracts obtained by Folch and ethanol-modified SC-CO2 extractions were similar. A sequential pure SC-CO2 and ethanol-modified SC-CO2 extraction was carried out to separate nonpolar lipids in the first fraction, thereby concentrating phospholipids in the second fraction. This sequential extraction produced a highly concentrated phospholipid extract (76%, wt/wt). To the best of our knowledge, this is the highest phospholipid concentration reported from buttermilk powder. Thus, this phospholipid-rich extract can be used in the development of functional foods as a food-grade emulsifier with potential health-promoting effects.

Application of supercritical carbon dioxide to enhance the aroma of whole sorghum flour for use in 3D printing of sorghum cookies.

Sorghum is a promising ingredient for new food products due to its high fiber content, slow digestibility, drought resistance, and gluten-free nature. One of the main challenges in sorghum-based products is the unpleasant aroma compounds found in grain sorghum. Therefore, in this study, sorghum flour was treated via supercritical carbon dioxide (SC-CO2) to remove undesired aroma compounds. The resulting SC-CO2-treated flours were used to generate dough for 3D food printing. At the optimized conditions, sorghum cookies were 3D-printed using 60 % water and a nozzle diameter of 1.5 mm. All dough samples produced with untreated and SC-CO2-treated sorghum flours exhibited shear-thinning behavior. Changing the treatment pressure (8-15 MPa) or temperature (40-60 °C) did not significantly affect the viscosity of the dough samples. Moreover, the sorghum cookie doughs had higher G' and G″ values after the SC-CO2 treatments (G' > G″). Doughs generated from flours treated at 15 MPa - 40 °C and 8 MPa - 60 °C showed lower adhesiveness compared to the ones produced from untreated flour, whereas 15 MPa - 60 °C treatment did not affect the adhesiveness. After baking, the 3D-printed cookies from SC-CO2-treated flour exhibited significantly lower redness (a*), but the hardness of the cookies was not affected by SC-CO2 treatment. Overall, the SC-CO2 treatment of sorghum flour did not negatively affect the quality parameters of the 3D-printed cookies while enhancing the aroma of the flour.

In vitro digestion of starch and protein aerogels generated from defatted rice bran via supercritical carbon dioxide drying.

This study investigated the in vitro digestibility of starch and protein aerogels produced from defatted rice bran (DRB), an underutilized rice processing byproduct, using supercritical carbon dioxide (SC-CO2) drying. The extracted starch (i.e., purified starch), crude starch, and proteins were used for the aerogel formation at 15% (w/w) concentration and further characterized. All aerogels exhibited three-dimensional open porous structures with high surface areas of 36-47 m2/g, densities lower than 0.3 g/cm3, and porosities higher than 84%. The starch hydrolyses in starch and crude starch aerogels were 86 and 73%, respectively, while the protein hydrolysis in protein aerogels reached up to 82% after sequential oral, gastric, and intestinal digestion. Thus, the hydrolysis rates achieved in simulated digestions suggest that the developed aerogels from DRB have the potential to serve as vehicles for delivering bioactive compounds and add value to the underutilized DRB.

Lutein encapsulation into dual-layered starch/zein gels using 3D food printing: Improved storage stability and in vitro bioaccessibility.

The ability of 3D printing to encapsulate, protect, and enhance lutein bioaccessibility was investigated under various printing conditions. A spiral-cube-shaped geometry was used to investigate the effects of printing parameters, namely zein concentration (Z; 20, 40, and 60 %) and printing speed (PS; 4, 8, 14, and 20 mm/s). Coaxial extrusion 3D printing was used with lutein-loaded zein as the internal flow material, and corn starch paste as the external flow material. The viscosities of the inks, microstructural properties, storage stability, and bioaccessibility of encapsulated lutein were determined. The sample printed with a zein concentration of 40 % at a printing speed of 14 mm/s (Z-40/PS-14) exhibited the best shape integrity. When lutein was entrapped in starch/zein gels (Z-40/PS-14), only 39 % of lutein degraded after 21 days at 25 °C, whereas 78 % degraded at the same time when crude lutein was studied. Similar improvements were also observed after storing at 50 °C for 21 days. Furthermore, after simulated digestion, the bioaccessibility of encapsulated lutein (9.8 %) was substantially higher than that of crude lutein (1.5 %). As a result, the developed delivery system using 3D printing could be an effective strategy for enhancing the chemical stability and bioaccessibility of bioactive compounds (BCs).

Upcycling imperfect broccoli and carrots into healthy snacks using an innovative 3D food printing approach.

Vegetables are healthy foods with nutritional benefits; however, nearly one-third of the world's vegetables are lost each year, and some of the losses happen due to the imperfect shape of the vegetables. In this study, imperfect vegetables (i.e., broccoli and carrots) were upcycled into freeze-dried powders to improve their shelf-life before they were formed into food inks for 3D printing. The rheology of the food inks, color analysis of the uncooked and cooked designs, and texture analysis of the cooked designs were determined. The inks with 50% and 75% vegetables provided the best printability and shape fidelity. 3D printing at these conditions retained a volume comparable to the digital file (14.4 and 14.3 cm3 vs. 14.6 cm3, respectively). The control, a wheat flour-based formulation, showed the lowest level of stability after 3D printing. The viscosity results showed that all the food inks displayed shear-thinning behavior, with broccoli having the greatest effect on viscosity. There was a significant color difference between uncooked and cooked samples, as well as between different formulations. The hardness of the baked 3D-printed samples was affected by the type and content of vegetable powders, where carrot-based snacks were notably harder than snacks containing broccoli. Overall, the results show that 3D food printing can be potentially used to reduce the loss and waste of imperfect vegetables.

Effect of a novel drying method based on supercritical carbon dioxide on the physicochemical properties of sorghum proteins.

The aim of this research was to use supercritical carbon dioxide (SC-CO2) drying as a novel approach for generating sorghum protein concentrates/isolates with enhanced functional properties. Sorghum protein extracts were obtained from white whole-grain sorghum flour and were dried by two methods, namely, freeze-drying and SC-CO2 drying. The collected proteins were characterized for their morphology, color, crystallinity, surface hydrophobicity, emulsifying activity index (EAI), creaming index (CI), foaming capacity (FC), foaming stability (FS), protein solubility, chemical interactions, and viscosity. The SC-CO2-dried proteins exhibited higher porosity compared to the freeze-dried ones with smaller particle sizes (∼5.1 vs. 0.4 µm, respectively). The XRD patterns indicated that the SC-CO2-dried proteins had a lower crystallinity than the freeze-dried proteins. However, the surface hydrophobicities of the freeze-dried and SC-CO2-dried proteins were similar. The EAI results showed that the emulsifying activity of freeze-dried protein powder (40.6) was better than that of SC-CO2-dried protein powder (29.8). Nevertheless, the solubility of SC-CO2-dried proteins was higher than that of freeze-dried proteins in most of the pHs investigated. Overall, the proposed SC-CO2 drying method has the potential to generate porous protein powders with improved solubility that can be used in developing functional foods.

Generation of porous starch beads via a 3D food printer: The effects of amylose content and drying technique.

A new approach via extrusion-based 3D food printing (3DP) was developed to fabricate porous spherical beads from corn starches with different amylose contents (i.e., 25, 55, and 72 %). The effects of amylose content and drying method, i.e., freeze-drying and supercritical carbon dioxide (SC-CO2), on the structural properties of the starch beads were investigated. The shape and size of the 3D-printed beads highly depended on the starches' amylose content as it affected the rheological properties of the inks. The smallest 3D-printed bead size was ∼980 µm generated from high amylose (72 %) corn starch. 3DP of starch with high amylose content along with SC-CO2 drying resulted in starch beads with superior properties. The SC-CO2-dried beads showed a significantly higher surface area (175 m2/g) than the freeze-dried ones (<1 m2/g).

Extraction of phenolic compounds from rice husk via ethanol-water-modified supercritical carbon dioxide.

Rice husk, a rice processing byproduct generated in large quantities (∼20% of the grain weight), creates a major disposal problem for the rice industry. However, rice husk contains high-value bioactive compounds that can provide potential health benefits. The objective of this study was to extract high-value phenolic compounds from rice husk using supercritical carbon dioxide (SC-CO2) technology. In this study, the effects of different extraction conditions, namely, temperature (40 and 60 °C), pressure (30 and 40 MPa), and ethanol concentration (15 and 25%, w/w) on the total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity (AA) were investigated. The extraction of phenolic compounds was also studied using different SC-CO2 modifiers, i.e., ethanol and ethanol-water. The highest TPC, TFC, and AA were achieved with 30 MPa, 60 °C, and 25% ethanol-water (50%, v/v) cosolvent mixture as 1.29 mg gallic acid equivalent (GAE)/g, 0.40 mg catechin equivalent (CE)/g, and 0.23 mg Trolox equivalent (TE)/g, respectively. Increasing water content up to 50% (v/v) in the cosolvent significantly improved the extraction yield. p-Coumaric, ferulic, and syringic acids were the predominant phenolic acids in the extracts obtained by cosolvent-modified SC-CO2 and methanol extractions. In addition, ethanol-water-modified SC-CO2 increased rice husk's porosity, which could be a potential pretreatment to enhance cellulose extraction. Thus, ethanol-water-modified SC-CO2 can be utilized to recover polar bioactive compounds from food processing byproducts for developing functional foods while eliminating the use of toxic organic solvents.

An extrusion-based 3D food printing approach for generating alginate-pectin particles.

In the present study, alginate-pectin (Al-P) hydrogel particles containing varied total gum concentrations (TGC) at a constant Al:P ratio of 80:20 were formed utilizing an innovative extrusion-based 3D food printing (3DFOODP) approach. The 3DFOODP conditions, namely, TGC (1.8, 2.0, and 2.2 wt%) and nozzle size (0.108, 0.159, and 0.210 mm) were investigated. The 3DFOODP approach was compared with the conventional bead formation method via a peristaltic pump. All Al-P printing inks exhibited a shear-thinning behavior. The increased apparent viscosity, loss and storage moduli were associated with the increase in the TGC. The size of the wet 3D-printed Al-P hydrogel particles ranged between 1.27 and 1.59 mm, which was smaller than that produced using the conventional method (1.44-1.79 mm). Freeze-dried Al-P particles showed a porous structure with reduced crystallinity. No chemical interaction was observed between alginate and pectin. This is the first report on generating Al-P-based beads using a 3DFOODP technique that can create delivery systems with high precision and flexibility.

Fabrication of Porous Spherical Beads from Corn Starch by Using a 3D Food Printing System.

This study introduces a 3D food printing approach to fabricate spherical starch beads with small sizes and high porosity for the first time. The results illustrated that 3D food printing could generate starch beads in different sizes depending on the nozzle diameter, printing pressure, and ink viscosity. The 3D-printed beads were characterized for their morphology, crystallinity, and textural properties, while the starch-based ink was analyzed for its rheological properties. A suitable printing was attained when viscosity was in the range of 1000-1200 Pa.s at a low shear rate (˂0.1 s-1). Among the starch concentrations (10-15%, w/w) investigated, 15% starch concentration provided the best control over the shape of the beads due to its high storage modulus (8947 Pa), indicating higher gel strength. At this condition, the starch beads revealed an average size of ~650 µm, which was significantly smaller than the beads produced with other starch concentrations (10 and 12.5%), and had a density of 0.23 g/cm3. However, at lower starch concentrations (10%), the beads were not able to retain their spherical shape, resulting in larger beads (812-3501 µm). Starch crystallinity decreased by gelatinization, and the starch beads exhibited a porous structure, as observed from their SEM images. Overall, 3D food printing can be an alternative approach to preparing porous beads for the delivery of bioactive compounds with high precision.

Enhancing the bioaccessibility of lycopene from tomato processing byproducts via supercritical carbon dioxide extraction.

Tomato peel and seed from tomato processing industry are treated as waste; however, they contain lycopene, a high-value bioactive compound. In this study, lycopene was extracted from tomato peel and seed using supercritical carbon dioxide (SC-CO2) and hexane, and the bioaccessibilities of lycopene in the SC-CO2- and hexane-extracted oleoresins were investigated for the first time. The (Z)-lycopene content of the SC-CO2-extracted oleoresin (69%) was higher than that of hexane-extracted oleoresin (45%). Separation of the insoluble fraction from the oleoresins increased the (Z)-lycopene contents of the SC-CO2- and hexane-extracted oil fractions to 80% and 49%, respectively. The bioaccessibility of total-lycopene in the oleoresins was increased by 3.3-fold via SC-CO2 extraction, which was attributed to higher (Z)-lycopene content, and small-sized uniform distribution of lycopene in the oleoresin. SC-CO2 extraction is not only a green method for extraction of bioactive compounds, but also has the potential to improve health benefits of bioactive compounds.

Physical and textural properties of functional edible protein films from soybean using an innovative 3D printing technology.

Increasing market demand for sustainable, environmentally friendly edible film materials has called for the development of new customizable production methods utilizing emerging technologies such as 3D printing. We hereby report a new method to generate functional edible soy protein isolate films prepared from three types of soybeans (AR-R11-7999, MO-S17-17168, and MO-S17-19874R) using an innovative 3D printing technology. The protein contents in AR-R11-7999, MO-S17-17168, and MO-S17-19874R soybean meals and their corresponding protein isolates were 40.0, 39.1, and 39.9; and 84.5, 84.7, and 87.3 % (w/w, dry basis), respectively. Response surface methodology was used to maximize the tensile and puncture strength and minimize the thickness of the 3D-printed edible films using protein concentration, plasticizer concentration (glycerol), and drying time as the independent variables. The optimized film production conditions were determined as soy protein concentration: 8.91%, plasticizer concentration: 3.00%, and drying time: 3.98 h with a desirability value of 0.7428. The optimized conditions were then successfully verified with the original soybean lot with a nonsignificant difference in physical properties. At the optimized conditions, the 3D-printed edible films using three soybean lots revealed: 0.108-0.114 mm thickness; 14.79-16.07 MPa tensile strength; 6.97-8.20 N puncture strength; 90.81-91.53, -1.89 to -1.31, and 14.85-17.25 were color parameters L*, a*, and b*, respectively; 1.22-1.36 g/cm3 density; and 104.4-105.7% elongation at break ratio (%). PRACTICAL APPLICATION: Edible soy protein films produced by an extrusion-based 3D printing approach are highly customizable and precise, and could be produced at an industrial scale. This newly produced environment-friendly soy protein-based edible film can serve as an alternate packaging to synthetic plastics and reduce the environmental landfill problem while adding value to soybean produced in the mid-south United States.

Liposomes for oral delivery of protein and peptide-based therapeutics: challenges, formulation strategies, and advances.

Throughout the past decade, there has been a rapid growth in the development of protein/peptide-based therapeutics. These therapeutics have found widespread applications in the treatment of cancer, infectious diseases, and other metabolic disorders owing to their several desirable attributes, such as reduced toxicity, diverse biological activities, high specificity, and potency. Most protein/peptide-based drugs are still administered parenterally, and there is an unprecedented demand in the pharmaceutical industry to develop oral delivery routes to increase patient acceptability and convenience. Recent advancements in nanomedicine discoveries have led to the development of several nano and micro-particle-based oral delivery platforms for protein/peptide-based therapeutics and among these, liposomes have emerged as a prominent candidate. Liposomes are spherical vesicles composed of one or more phospholipid bilayers enclosing a core aqueous phase. Their unique amphiphilic nature enables encapsulation of a diverse range of bioactives/drugs including both hydrophobic and hydrophilic compounds for delivery. Against this backdrop, this review provides an overview of the current approaches and challenges associated with the routes and methods of oral administration of protein/peptide-based therapeutics by using liposomes as a potential vehicle. First, the conventional and innovative liposome formation approaches have been discussed along with their applications. Next, the challenges associated with current approaches for oral delivery of protein and peptide-derived therapeutics have been thoroughly addressed. Lastly, we have critically reviewed the potential of liposomes utilization as vehicles for oral delivery of proteins emphasizing the current status and future directions in this area.

Heat stability of emulsions using functionalized milk protein concentrate generated by supercritical fluid extrusion.

In this study, thermostable oil-in-water emulsions containing high protein contents were developed using milk protein concentrate (MPC) that was functionalized by supercritical fluid extrusion (SCFX) processing at low temperature and shear. Functionalized MPC (f-MPC) emulsions (3% protein-80% oil and 10% protein-50% oil) were compared with emulsions stabilized by commercial MPC (c-MPC), sodium caseinate (NaCas), and a commercial mayonnaise for their emulsifying properties and heat stability at 70 and 90 °C for 30 min, and 121 °C for 15 min. Zeta-potentials and interfacial protein concentrations of f-MPC emulsions were higher than that of c-MPC emulsions. f-MPC emulsions remained stable against creaming for at least 8 weeks at room temperature (23 °C), while their c-MPC counterparts showed significant creaming at the same conditions. Even after heating at 121 °C for 15 min, f-MPC emulsions retained their structural integrity as observed from their confocal images, droplet size distributions, and viscosities. In contrast, c-MPC emulsions and mayonnaise disintegrated upon heating at 121 °C for 15 min, and oil droplets of mayonnaise partially coalesced during heating at 90 °C for 30 min. f-MPC emulsions revealed higher viscosities compared to c-MPC emulsions, providing them improved stability. Viscosities of f-MPC emulsions were not significantly affected by heating at 90 °C for 30 min, while other emulsions exhibited a substantial increase in their viscosities due to protein denaturation and aggregation. Thus, f-MPC emulsions can be utilized in the development of protein-enriched functional foods (e.g., spreads) that are stable against high heat treatments.

A novel and green nanoparticle formation approach to forming low-crystallinity curcumin nanoparticles to improve curcumin's bioaccessibility.

Health-promoting effects of curcumin are well-known; however, curcumin has a very low bioavailability due to its crystalline structure. The main objective of this study was to develop a novel green nanoparticle formation method to generate low-crystallinity curcumin nanoparticles to enhance the bioavailability of curcumin. Nanoporous starch aerogels (NSAs) (surface area of 60 m2/g, pore size of 20 nm, density of 0.11 g/cm3, and porosity of 93%) were employed as a mold to produce curcumin nanoparticles with the help of supercritical carbon dioxide (SC-CO2). The average particle size of the curcumin nanoparticles was 66 nm. Impregnation into NSAs decreased the crystallinity of curcumin and did not create any chemical bonding between curcumin nanoparticles and the NSA matrix. The highest impregnation capacity was 224.2 mg curcumin/g NSA. Curcumin nanoparticles significantly enhanced the bioaccessibility of curcumin by 173-fold when compared to the original curcumin. The concentration of curcumin in the bioaccessible fraction was improved from 0.003 to 0.125 mg/mL by impregnation of curcumin into NSAs (42-fold). This is a novel approach to produce food grade curcumin nanoparticles with reduced crystallinity and maximize the utilization of curcumin due to increased bioaccessibility.

In vitro bioaccessibility of novel low-crystallinity phytosterol nanoparticles in non-fat and regular-fat foods.

Crystalline structure of phytosterols leads to poor bioavailability and makes their incorporation into foods challenging. Bioaccessibility of first-of-their-kind low-crystallinity phytosterol nanoparticles impregnated in nanoporous starch aerogels (PS-NSA) was evaluated in non-, low-, and regular-fat solid and aqueous food formulations, namely, granola bars and puddings for the first time. Bioaccessibility of the phytosterol nanoparticles was significantly higher than that of crude phytosterols in all food formulations (p < .05); it was 88.2 and 91.8% for low- or regular-fat granola bars, respectively, whereas bioaccessibility of crude phytosterols was ca. 30% in those formulations. However, decreasing the lipid content to zero resulted in lower phytosterols' bioaccessibilities from both PS-NSA (53%) and crude phytosterols (ca. 16%) in non-fat granola bars. Bioaccessibility of crude phytosterols (2%) was significantly enhanced with PS-NSA (19%) in the pudding formulation. PS-NSA allows preparation of low- and non-fat foods enriched with phytosterols while enhancing the health benefits of phytosterols with smaller doses.

In Vitro Bioaccessibility of Low-Crystallinity Phytosterol Nanoparticles Generated Using Nanoporous Starch Bioaerogels.

Phytosterols are natural health-promoting bioactive compounds; however, phytosterols have very limited bioavailability due to their crystalline lipophilic structure. With the aim of improving bioaccessibility, low-crystallinity phytosterol nanoparticles were generated by supercritical carbon dioxide (SC-CO2 ) impregnation of phytosterols into nanoporous starch aerogels (NSAs). The in vitro bioaccessibility of the phytosterol nanoparticles (35%) was significantly higher than that of the crude phytosterols (3%) after sequential oral, gastric, and intestinal digestion. The percentages of starch hydrolysis were not different among the various NSA preparations and reached to 64% after sequential digestion. The zeta potential of the phytosterol nanoparticles was higher compared to that of crude phytosterols in the micellar phase; indicating higher stability. The findings of this study support the use of NSA to produce nanoparticles of reduced crystallinity to improve the bioaccessibility of the lipophilic bioactive compounds. PRACTICAL APPLICATIONS: This novel process can decrease the size and crystallinity of phytosterols and thus improve phytosterols' bioavailability. It is a blueprint to apply to other water insoluble food bioactives. This novel approach may (i) improve the health benefits of water-insoluble bioactives; (ii) enable food manufacturers to add water-insoluble bioactives into low- and high-fat foods to produce health-promoting foods; and (iii) enhance the cost-benefit ratio of water insoluble bioactives.