A sodium alginate gel bead adsorbent doping with amidoxime-modified hydroxyapatite for the efficient adsorption of uranium.

In this work, the modified­sodium alginate gel beads were prepared by sol-gel method. Due to the presence of water channels in the sodium alginate gel bead, amidoxime groups and PO43- were exposed to the surface of the adsorbent to the maximum extent, resulting in the excellent adsorption capacity of modified­sodium alginate gel beads. The introduction of amidoxime-modified hydroxyapatite significantly improved the adsorption capacity and the adsorption rate of the gel beads. The adsorption capacity increased from 308.7 to 466.0 mg/g and the adsorption equilibrium time was shortened from 300 min to 120 min. The modified­sodium alginate gel bead possessed the advantages of short adsorption time, high adsorption efficiency and large adsorption capacity, which could be regarded as a potential adsorbent for uranium. Moreover, the uranium removal ability on the modified gel beads was mainly attributed to the Coulomb force between PO43- and uranium and the complexation between uranium and amidoxime groups. In summary, this work would provide a new idea for the modification and application of sodium alginate-based materials.

Development of Anthocyanin-Rich Gel Beads from Colored Rice for Encapsulation and In Vitro Gastrointestinal Digestion.

Colored rice anthocyanins are water-soluble natural pigments that can be used as an active ingredient in healthy food and pharmaceutical products. However, anthocyanin utilization is limited because of its instability. This work produced anthocyanin-rich gel beads from colored rice using a modified ionotropic gelation technique for encapsulation, and their efficacy was studied in vitro in the gastrointestinal tract. In total, 15 colored rice samples of three types (whole grain rice, ground rice, and ground germinated rice) were screened to identify the highest anthocyanin content. The anthocyanin content of the whole grain rice was significantly (p < 0.05) higher than it was in the ground and ground germinated rice. The sample with the highest anthocyanin content (1062.7 µg/g) was the black glutinous rice grain from Phrae, chosen based on its anthocyanin-rich crude extract. A new formula using a modified ionotropic gelation technique was prepared for the inclusion of the extract in gel beads. The results indicated that the incorporation of oil and wax significantly increased the encapsulation efficiency of the gel beads (% EE value of 85.43%) and improved the bioavailability of the active ingredient. Moreover, after simulated digestion, the release of anthocyanin and total phenolic content occurred more than five times. Scanning electron microscopy revealed that the surface of the gel beads was smooth. Furthermore, the presence of polyphenols and polysaccharides in the gel beads was confirmed using FTIR. The oil-wax-incorporated, anthocyanin-rich gel beads could be implemented for antioxidant delivery into the gastrointestinal tract to further improve healthy food and nutraceutical products.

Hybrid biodegradable materials from starch and hydrocolloid: fabrication, properties and applications of starch-hydrocolloid film, gel and bead.

The potential for utilizing starch and hydrocolloids as sustainable biomaterials has garnered significant attention among researchers. The biodegradability and functional properties of composite films, gels, and beads, as well as their environmental friendliness, make them attractive options for a variety of applications. However, the hydrophilicity, brittleness, and regeneration limitations of starch materials can be addressed through the incorporation of non-starch hydrocolloids. This article summarizes the formation mechanisms and interactions of starch-hydrocolloid films, gels, and gel beads, evaluates the factors that affect their structural and functional properties, and presents an overview of the progress made in their physicochemical and functional applications. The structure of starch-hydrocolloid composites is primarily formed through hydrogen bond interactions, and the source, proportion, and preparation conditions of the components are critical factors that affect the properties of the biomaterials. Starch-hydrocolloid films are primarily used for extending the shelf life of food products and detecting food freshness. Starch-hydrocolloid gels are utilized as adsorption materials, wound dressings, and flexible sensors, and starch-hydrocolloid beads are primarily employed for the controlled release of bioactive substances. It is clear that starch-hydrocolloid composites have the potential to develop novel advanced materials for various applications in the food, biological, and materials industries.

Highly Stable, Mechanically Enhanced, and Easy-to-Collect Sodium Alginate/NZVI-rGO Gel Beads for Efficient Removal of Cr(VI).

Nanoscale zero-valent iron (NZVI) is a material that is extensively applied for water pollution treatment, but its poor dispersibility, easy oxidation, and inconvenient collection limit its application. To overcome these drawbacks and limit secondary contamination of nanomaterials, we confine NZVI supported by reduced graphene oxide (rGO) in the scaffold of sodium alginate (SA) gel beads (SA/NZVI-rGO). Scanning electron microscopy showed that the NZVI was uniformly dispersed in the gel beads. Fourier transform infrared spectroscopy demonstrated that the hydrogen bonding and conjugation between SA and rGO allowed the NZVI-rGO to be successfully embedded in SA. Furthermore, the mechanical strength, swelling resistance, and Cr(VI) removal capacity of SA/NZVI-rGO were enhanced by optimizing the ratio of NZVI and rGO. Interestingly, cation exchange may drive Cr(VI) removal above 82% over a wide pH range. In the complex environment of actual Cr(VI) wastewater, Cr(VI) removal efficiency still reached 70.25%. Pseudo-first-order kinetics and Langmuir adsorption isotherm are preferred to explain the removal process. The mechanism of Cr(VI) removal by SA/NZVI-rGO is dominated by reduction and adsorption. The sustainable removal of Cr(VI) by packed columns could be well fitted by the Thomas, Adams-Bohart, and Yoon-Nelson models, and importantly, the gel beads maintained integrity during the prolonged removal. These results will contribute significant insights into the practical application of SA/NZVI-rGO beads for the Cr(VI) removal in aqueous environments.

Tannic acid-assisted fabrication of antibacterial sodium alginate-based gel beads for the multifunctional adsorption of heavy metal ions and dyes.

The existence of heavy metals and dyes seriously affects the ecological environment and human safety. Antibacterial adsorption materials with the broad-spectrum removal of multiple pollutants are urgently required for water remediation. Herein, a sustainable and antibacterial sodium alginate (SA) gel bead adsorbent with honeycomb cellular architecture is developed by the biomimetic deposition polyphenolic tannic acid (TA) induced grafting diethylenetriamine (DETA) under mild conditions for efficient removal of Cr(VI) and dyes. Taking advantage of the catechol surface chemistry, TA occurring rapid polymerization with DETA monomers not only enhances the water resistance and thermal stability of the gel bead, but also introduces abundant polyphenolic functional groups and active adsorption sites. The multifunctional gel bead showed outstanding antibacterial activity against S. aureus (sterilization rates: 83.8 %) and E. coli (sterilization rates: 99.5 %). The maximum adsorption capacity of gel bead for Cr(VI) was 163.9 mg/g. Moreover, the removal efficiency of the gel bead for dyes of Safranine T and Rhodamine B was 89.5 % (maximum adsorption capacity: 537 mg/g) and 76.7 % (maximum adsorption capacity: 460.2 mg/g), respectively, indicating its excellent broad-spectrum adsorption performance for multiple pollutants. Therefore, TA-assisted fabrication of SA-based gel bead with excellent antibacterial property is a promising multifunctional adsorption material for practical water remediation.

Fabrication of phosphoric-crosslinked chitosan@g-C3N4 gel beads for uranium(VI) separation from aqueous solution.

In this work, a novel g-C3N4 filled, phosphoric-crosslinked chitosan gel bead (P-CS@CN) was successfully prepared to adsorb U(VI) from water. The separation performance of chitosan was improved by introducing more functional groups. At pH 5 and 298 K, the adsorption efficiency and adsorption capacity could reach 98.0 % and 416.7 mg g-1, respectively. After adsorption, the morphological structure of P-CS@CN did not change and adsorption efficiency remained above 90 % after 5 cycles. P-CS@CN exhibited an excellent applicability in water environment based on dynamic adsorption experiments. Thermodynamic analyses demonstrated the value of ΔG, manifesting the spontaneity of U(VI) adsorption process on P-CS@CN. The positive values of ΔH and ΔS showed that the U(VI) removal behavior of P-CS@CN was an endothermic reaction, indicating that the increase of temperature was great benefit to the removal. The adsorption mechanism of P-CS@CN gel bead could be summarized as the complexation reaction with the surface functional groups. This study not only developed an efficient adsorbent for the treatment of radioactive pollutants, but also provided a simple and feasible strategy for the modification of chitosan-based adsorption materials.

Universal Way to "Glue" Capsules and Gels into 3D Structures by Electroadhesion.

We demonstrate the use of electroadhesion (EA), i.e., adhesion induced by an electric field, to connect a variety of soft materials into 3D structures. EA requires a cationic and an anionic material, but these can be of diverse origin, including covalently cross-linked hydrogels made by polymerizing charged monomers or physical gels/capsules formed by the ionic cross-linking of biopolymers (e.g., alginate and chitosan). Between each cationic/anionic pair, EA is induced rapidly (in ∼10 s) by low voltages (∼10 V DC)─and the adhesion is permanent after the field is turned off. The adhesion is strong enough to allow millimeter-scale capsules/gels to be assembled in 3D into robust structures such as capsule-capsule chains, capsule arrays on a base gel, and a 3D cube of capsules. EA-based assembly of spherical building blocks can be done more precisely, rapidly, and easily than by any alternative techniques. Moreover, the adhesion can be reversed (by switching the polarity of the field)─hence any errors during assembly can be undone and fixed. EA can also be used for selective sorting of charged soft matter─for example, a 'finger robot' can selectively 'pick up' capsules of the opposite charge by EA and subsequently 'drop off' these structures by reversing the polarity. Overall, our work shows how electric fields can be used to connect soft matter without the need for an adhesive or glue.

Analysis of mRNA Subcellular Distribution in Collective Cell Migration.

The movement of groups of cells by collective cell migration requires division of labor between group members. Therefore, distinct cell identities, unique cell behaviors, and specific cellular roles are acquired by cells undergoing collective movement. A key driving force behind the acquisition of discrete cell states is the precise control of where, when, and how genes are expressed, both at the subcellular and supracellular level. Unraveling the mechanisms underpinning the spatiotemporal control of gene expression in collective cell migration requires not only suitable experimental models but also high-resolution imaging of messenger RNA and protein localization during this process. In recent times, the highly stereotyped growth of new blood vessels by sprouting angiogenesis has become a paradigm for understanding collective cell migration, and consequently this has led to the development of numerous user-friendly in vitro models of angiogenesis. In parallel, single-molecule fluorescent in situ hybridization (smFISH) has come to the fore as a powerful technique that allows quantification of both RNA number and RNA spatial distribution in cells and tissues. Moreover, smFISH can be combined with immunofluorescence to understand the precise interrelationship between RNA and protein distribution. Here, we describe methods for use of smFISH and immunofluorescence microscopy in in vitro angiogenesis models to enable the investigation of RNA and protein expression and localization during endothelial collective cell migration.

Guidance on Processing the 10x Genomics Single Cell Gene Expression Assay.

The demand for technologies that allow the study of gene expression at single cell resolution continues to increase. One such assay was launched in 2016 by the US-based company 10x Genomics Inc. Utilizing the power of the single cell on a large scale (Zheng et al. Nat Commun 8:14049, 2017)-capturing thousands of cells at once-has shaped life sciences ever since and allowed researchers to discover new insights within their respective fields of study such as oncology, neurobiology, and immunology (among others). Obtaining high-data quality is the key to being able to make these meaningful discoveries, which in turn is directly linked to the quality of the initial cell (or nuclei) suspension that is used to load the 10x Genomics Chromium Single Cell Gene Expression assay. A successful workflow relies on a cell suspension which is fully dissociated, extremely clean, and of high viability. While the workflow itself has been detailed elsewhere (De Simone et al. Methods Mol Biol 1979:87-110, 2019), in this chapter we will focus on the importance of the quality of the initial cell suspension, as well as common mistakes that can occur while running a Single Cell Gene Expression assay. The descriptions of these tips and tricks refer to the current version of the 10x Genomics User Guide (Chromium Single Cell 3' Reagent Kits User Guide (v3.1 Chemistry Dual Index). https://support10xgenomicscom/single-cell-gene-expression/index/doc/user-guide-chromium-single-cell-3-reagent-kits-user-guide-v31-chemistry-dual-index) which can be downloaded from the Support section on the 10x Genomics website (10x Genomics website. https://www10xgenomicscom). These documents and user guides are continuously improved and updated; hence, it is important to regularly check the company's website for the most recent version.

Optimization and release characteristics of catechin-loaded calcium pectinate beads by internal gelation.

The aim of this study was to optimize the composition of catechin-loaded calcium pectinate gel (CPG) beads formed using internal gelation and to evaluate the sustained-release behavior of catechin. The pectin concentration, catechin-to-pectin ratio, and calcium carbonate-to-pectin ratio were optimized for the sustained catechin release (2.89, 28.92, and 32.79%, respectively). The catechin release profiles were analyzed using the simple enzyme kinetic-like semi empirical model newly proposed in this study. The actual release rate was found to be the fastest in the simulated intestinal fluid (SIF), followed by the simulated gastric fluid (SGF) and the pH 4.5 buffer, whereas the thermodynamic equilibrium was achieved fastest in the pH 4.5 buffer, followed by SGF and SIF. Glutaraldehyde treatment suppressed catechin release in all tested media. These results suggest that internally gelled CPG beads are suitable for catechin delivery, and crosslinkers, such as glutaraldehyde, can effectively sustain their release.

Fabrication and characterization of phycocyanin-alginate-pregelatinized corn starch composite gel beads: Effects of carriers on kinetic stability of phycocyanin.

Composite gel beads using calcium alginate and different concentrations of pregelatinized corn starch (PCS) were produced to encapsulate phycocyanin (PC). Rheological properties of different sodium alginate/PCS/PC mixtures, structural and morphological properties of beads, and kinetic stability of encapsulated PC (upon heating at various time-temperature combinations) were then assessed. Rheological properties of the mixtures exhibited shear thinning behaviors. Aquagram revealed that the PC-containing beads had more water structure with weak­hydrogen bonds. Morphological images represented less subsidence in the structures of composite gel beads, unlike PCS-free beads. Kinetic study showed that degradation rate constant values of PC encapsulated in composite gel beads (1.08-3.45 × 10-4, 3.38-4.43 × 10-4, and 5.57-15.32 × 10-4 s-1) were lower than those in PCS-free alginate gel beads (4.45 × 10-4, 9.20 × 10-4, and 18.04 × 10-4 s-1) at 40, 50, and 60 °C, respectively. This study suggests that the composite gel beads can improve PC stability.

Customization of liquid-core sodium alginate beads by molecular engineering.

Detailed alginate molecular structural factors that modulated the texture of liquid core alginate beads were investigated. Under the same CaCl2 concentration and contacting time, a higher bursting force can be generated from both high Mw and low M/G alginate beads. G-block took part in faster dimerization, producing a compact gel network and long chains providing an elastic response generated by interchain interaction. Within same bursting force, high Mw beads were elastic than high G ones. Shorter chain length and higher M residue allowed more water to trap in the gel network, causing accretion in thickness and downed gel strength. The lowest shell thickness (0.11±0.01mm) was measured from the highest Mw and M/G sample, while the thickest (0.78±0.08mm) was from the lowest Mw and M/G alginate. The beads became brittle with a shortened chain length. Thus, alginate-based food products with a desired textural profile can be designed by varying alginate structures.

Understanding the Drying Behavior of Regenerated Cellulose Gel Beads: The Effects of Concentration and Nonsolvents.

The drying behavior of regenerated cellulose gel beads swollen with different nonsolvents (e.g., water, ethanol, water/ethanol mixtures) is studied in situ on the macroscopic scale with an optical microscope as well as on nanoscale using small-angle/wide-angle X-ray scattering (SAXS/WAXS) techniques. Depending on the cellulose concentration, the structural evolution of beads during drying follows one of three distinct regimes. First, when the cellulose concentration is lower than 0.5 wt %, the drying process comprises three steps and, regardless of the water/ethanol mixture composition, a sharp structural transition corresponding to the formation of a cellulose II crystalline structure is observed. Second, when the cellulose concentration is higher than 5.0 wt %, a two-step drying process is observed and no structural transition occurs for any of the beads studied. Third, when the cellulose concentration is between 0.5 and 5.0 wt %, the drying process is dependent on the nonsolvent composition. A three-step drying process takes place for beads swollen with water/ethanol mixtures with a water content higher than 20%, while a two-step drying process is observed when the water content is lower than 20%. To describe the drying behavior governed by the cellulose concentration and nonsolvent composition, a simplified phase diagram is proposed.

Macro- and Microstructural Evolution during Drying of Regenerated Cellulose Beads.

The macro- and microstructural evolution of water swollen and ethanol swollen regenerated cellulose gel beads have been determined during drying by optical microscopy combined with analytical balance measurements, small-angle X-ray scattering (SAXS), and wide-angle X-ray scattering (WAXS). Two characteristic length scales, which are related to the molecular dimension of cellulose monomer and elongated aggregates of these monomers, could be identified for both types of beads by SAXS. For ethanol swollen beads, only small changes to the structures were detected in both the SAXS and WAXS measurements during the entire drying process. However, the drying of cellulose from water follows a more complex process when compared to drying from ethanol. As water swollen beads dried, they went through a structural transition where elongated structures changed to spherical structures and their dimensions increased from 3.6 to 13.5 nm. After complete drying from water, the nanostructures were characterized as a combination of rodlike structures with an approximate size of cellulose monomers (0.5 nm), and spherical aggregates (13.5 nm) without any indication of heterogeneous meso- or microporosity. In addition, WAXS shows that cellulose II hydrate structure appears and transforms to cellulose II during water evaporation, however it is not possible to determine the degree of crystallinity of the beads from the present measurements. This work sheds lights on the structural changes that occur within regenerated cellulose materials during drying and can aid in the design and application of cellulosic materials as fibers, adhesives, and membranes.

Encapsulation and release behavior of curcumin based on nanoemulsions-filled alginate hydrogel beads.

To provide the bilateral advantages of emulsions and hydrogels, a facile approach was used to fabricate nanoemulsions filled hydrogel beads through combining the method of self-emulsification and sodium alginate (SA) ionic gelation. The encapsulation and release behavior of curcumin (Cur) were further investigated. The results indicated that Cur packaged nanoemulsions were with the size of 24.26 ±â€¯0.22 nm. The nanoemulsions filled SA hydrogel beads were spherical shell with the diameter of 0.46 ±â€¯0.02 mm. For Cur, the EE and LC of emulsion filled SA hydrogel beads were 99.15 ±â€¯0.85% and 7.25 ±â€¯3.16 mg/g respectively. The release behavior could be regulated by external pH condition. The release behavior at pH 9.0 displayed a higher release rate than that at pH 7.0. Cur released behavior well followed the Hixcon-Crowell model which indicated that Cur was released in a diffusion-controlled model. Comparatively investigation of microstructure using field emission scanning electron microscope (FE-SEM) further investigates the corrosion behavior of SA gel beads during Cur release. The worth-while endeavor provided a practical combined technique of emulsions and ionic gelation to fabricate hybrid hydrogel beads that have potential in delivery system for hydrophobic composition.

Magnetic chitosan/sodium alginate gel bead as a novel composite adsorbent for Cu(II) removal from aqueous solution.

Using sodium alginate hydrogel as skeleton, in combination with chitosan and magnetic Fe3O4, a new type of magnetic chitosan/sodium alginate gel bead (MCSB) was prepared. Adsorptive removal of Cu(II) from aqueous solutions was studied by using the MCSB as a promising candidate in environmental application. Different kinetics and isotherm models were employed to investigate the adsorption process. Based on Fourier transform infrared spectroscopy, field-emission scanning electron microscope, CHNS/O elements analysis, vibration magnetometer, and various means of characterization, a comprehensive analysis of the adsorption mechanism was conducted. The MCSB had a good magnetic performance with a saturation magnetization of 12.5 emu/g. Elemental analysis proved that the addition of chitosan introduced a considerable amount of nitrogen-rich groups, contributing significantly to copper adsorption onto gel beads. The contact time necessary for adsorption was optimized at 120 min to achieve equilibrium. Experimental data showed that the adsorption process agreed well with the Langmuir isotherm model and the pseudo-second-order kinetics model. The theoretical maximum adsorption capacity of MCSB for Cu(II) could reach as high as 124.53 mg/g. In conclusion, the MCSB in this study is a novel and promising composite adsorbent, which can be applied for practical applications in due course.

Preparation and characterization of emulsion-filled gel beads for the encapsulation and protection of resveratrol and α-tocopherol.

Proteins and polysaccharides have been widely used as wall materials to prepare novel carriers for the encapsulation of a single bioactive component or co-encapsulation of different bioactive components. Emulsion-filled gel beads generally contain three essential components: oil droplets, interface membrane of emulsifier, and gel matrix. In this study, emulsion-filled gel beads were prepared by the combination of heat-denatured whey protein isolate (WPI)'s emulsification and alginate's gelation at various protein and oil contents, in order to discuss simultaneous encapsulation, protection and release of hydrophobic α-tocopherol and amphiphilic resveratrol. High efficiency for the co-encapsulation of resveratrol and α-tocopherol was achieved for the emulsion-filled gel beads containing 1% WPI and 1%-4% oil. WPI content was more important than the content of oil for the protection of resveratrol while the content of oil was more important for the encapsulation and protection of α-tocopherol during storage. The release of α-tocopherol was <20% while resveratrol was rapidly released during digestion. Resveratrol remained 73% after released from the emulsion-filled gel beads containing 1% WPI and 1%-2% oil for 6 h. These results should be useful for the development of biopolymer-based carriers for the delivery of different bioactive components.

Condensation of Counterions Gives Rise to Contraction Transitions in a One-Dimensional Polyelectrolyte Gel.

The equilibrium volume of a polyelectrolyte gel results from a balance between the tendency to swell caused by outbound polymer/counterion diffusion along with Coulomb interactions on the one hand; and, on the other, the elastic resilience of the cross-linked polymer network. Direct Coulomb forces contribute both to non-ideality of the equilibrated Donnan osmotic pressure, but also to stretching of the network. To isolate the effect of polyelectrolyte expansion, we have analyzed a "one-dimensional" version of a gel, a linear chain of charged beads connected by Hooke's law springs. As in the range of weak Coulomb strengths previously studied, the springs are significantly stretched by the repulsive interactions among the beads even when the Coulomb strength is strong enough to cause condensation of counterions. There is a quasi-abrupt transition from a stretched state to a partially collapsed state in a transition range between weak and strong Coulomb strengths. Fluctuations between stretched and contracted conformations occur within the transition range. As the solvent quality decreases past the transition range, a progressive collapse can result if the condensed counterions strengthen the spring constant.

Effects of gelling bath on the physical properties of alginate gel beads and the biological characteristics of entrapped HepG2 cells.

Optimizing alginate gel beads is necessary to support the survival, proliferation, and function of entrapped hepatocytes. In this study, gelling bath was modified by decreasing calcium ion concentration and increasing sodium ion concentration. Alginate gel beads (using 36% G sodium alginate) prepared in the modified gelling bath had more homogeneous structure and better mass transfer properties compared with the traditional gelling bath that contains only calcium ions. Moreover, alginate gel beads generated in the modified gelling bath could significantly promote the HepG2 cell proliferation and the growth of cell spheroids, and maintain the albumin secretion ability similar to alginate gel beads prepared in the traditional gelling bath with only calcium ions. The mass transfer properties and cell proliferation were similar in ALG beads with different M/G ratio (36% G and 55% G) generated in the modified gelling bath, whereas they were significantly increased compared with alginate gel beads (55% G) in traditional gelling bath. These results indicated that adjusting the gelling bath was a simple and convenient method to enhance the mass transfer properties of alginate gel beads for 3D hepatocyte culture, which might provide more hepatocytes for the bioartificial liver support system.

Novel decellularized liver matrix-alginate hybrid gel beads for the 3D culture of hepatocellular carcinoma cells.

Developing reliable three-dimensional (3D) cell culture systems that can mimic native tumor microenvironments is necessary for investigating the mechanism of hepatocellular carcinoma (HCC) metastasis and screen therapeutic drugs. In the present study, we developed decellularized liver matrix-alginate (DLM-ALG) hybrid gel beads. DLM powder was prepared by optimized decellularization methods and liquid nitrogen grinding. DLM-ALG beads were generated by dropping alginate solution containing DLM powder into a gelling bath. DLM powder concentration in alginate solution was ≤1% (w/v) and had no effect on the sphericity and mechanical stability of the beads. In addition, HCCLM3 cells cultured in 1% (w/v) DLM-ALG beads presented gradually enhanced viability during in vitro culture. The protein expression of urokinase plasminogen activator system and activity of matrix metalloproteinases (MMPs) of HCCLM3 cells, including MMP2 and MMP9, were more significantly promoted in DLM-ALG beads compared with that in conventional ALG beads without DLM powder. Moreover, the dose-dependent increase in HCCLM3 cell MMP activities was observed along with the DLM powder concentration in 0.5% and 1% DLM-ALG groups. Therefore, DLM-ALG beads might serve as a novel 3D culture system for exploring the mechanisms of HCC metastasis and screening therapeutic drugs.