Self-assembly Of Poly Research Articles

Nanoengineering Spikey Surfaces: Investigation of Reversible Organizational Control of Surface-Tethered Polypeptide Brushes.

Nature serves as an important source of inspiration for the innovation and development of micro- and nanostructures for advanced functional surfaces and substrates. One example used in nature is a spikey surface ranging from micrometer-sized spikes on pollen grains down to the nanometer-scale protein spikes found on viruses. This study explored the realization of such highly textured surfaces via the nanoengineering of self-assembled poly(γ-benzyl-l-glutamate) "nanospikes", exploiting solvent-induced chain organization, controlled surface chemical functionality, and enhanced stability in the form of polymer brushes. The reversible solvent-responsive behavior of these polymer chains and the aggregation behavior of the chain-ends were investigated via fluorescence characterization and studied through molecular simulations. Vapor-based solvent treatments were developed for orientation control with in situ analysis to understand film response and brush organizational behavior under different selected conditions. The effect of sub-100 nm nanopatterning on surface morphology and chain organization was examined via an integrated approach of experimental and computational studies. The methodologies established in this study present opportunities for engineering sophisticated nanoscale spikey surfaces with high customizability by means of nanolithography combined with solvent-assisted treatments.

Self-assembled nanoflower-mediated interfacial polymerization through tunable intra- and inter-layer channels to boost total heat exchange performance

Total heat exchange membranes (THEMs) are vital for minimizing energy consumption and enhancing indoor air quality in energy recovery ventilation (ERV) systems. Manipulating the surface morphology of polyamide (PA) membranes via nanofiller-mediated interfacial polymerization is key to advancing total heat recovery performance. This study presented the fabrication of PA thin-film nanocomposite (TFN) membranes by integrating self-assembled poly(amic acid-imide) nanoflowers (P(AA-I)-NFs) with intertwined nanosheets into the organic phase. This facile approach effectively eliminated nonselective interphase voids and defects, owing to the superior polymer affinity of the organic nanoflowers. The finely tuned intra- and inter-layer channels within P(AA-I)-NFs significantly impacted monomer mass transfer, facilitated the shuttle effect, expanded the interfacial polymerization zone, and led to the formation of a rougher, thicker PA layer with enhanced surface area. The optimized P(AA-I)-NFs/PA TFN membranes exhibited outstanding performance, including CO₂ permeability of 0.51 GPU, water vapor permeability of 656.59 GPU, and an enthalpy exchange efficiency of 71.47 %, surpassing the trade-off limitations typically observed in commercial and other advanced THEMs. These findings underscored the potential of P(AA-I)-NFs-mediated TFN membranes as highly competitive candidates for next-generation ERV systems.

Synthesis and Self-Assembly of Poly(4-acetoxystyrene) Cubosomes.

Polymer cubosomes (PCs) are a recent class of self-assembled nanostructures with great application potential due to their high porosity and surface area. Currently, most reported PCs consist of polystyrene block copolymers (BCPs), for which self-assembly parameters are rather well understood. Changing the block chemistry would be desirable to introduce more functionality; however, knowledge of adapting the self-assembly process to new chemistries remains limited. This work, reports on synthesizing poly(ethylene oxide)-block-poly(4-acetoxystyrene) and its copolymers with styrene, and provide conditions for their self-assembly into PCs with high yield and high inner order. It is shown that the polarity of the starting solvent toward the corona block allows tuning of the final morphology by controlling the corona volume and packing parameter.

Individual and simultaneous encapsulation and delivery of incompatible dyes in biocompatible multicompartment terpolymer micelles

Multicompartment micelles (MCMs) can deliver incompatible drug payloads because MCMs derive from the self-assembly of a terpolymer containing one hydrophilic block and two incompatible solvophobic blocks. In particular, MCMs resulting from the self-assembly of poly((sulfamate-carboxylate)isoprene)-block-polystyrene-block-poly(ethylene oxide) (PISC-b-PS-b-PEO) undergo irreversible transition to regular micelles at an alkaline pH. However, several aspects of these MCMs remain unexplored, such as the chemical transformation of PISC-b-PS-b-PEO into an amino-acid functional terpolymer, potential applications of these nanoparticles as dual drug delivery systems and their biocompatibility. In exploring these opportunities, we found that more than 90% of N-S bonds of sulfamate groups (of the PISC block) are cleaved upon acidic treatment of PISC-b-PS-b-PEO terpolymers, which are chemically transformed into a new polyzwitterion triblock terpolymer, that is, poly((amino-carboxylate)isoprene)-block-polystyrene-block-poly(ethylene oxide) (PACIS-b-PS-b-PEO), but retain their MCM morphology. Into these MCMs, two incompatible (Nile Red (hydrophobic) and eosin (hydrophilic)) guest molecules can be loaded separately and together in the predesigned cores without significantly disturbing the structure, as shown by cryo-TEM and light scattering. In addition to their high stability, PACIS-b-PS-b-PEO MCMs are also pH-responsive, whether loaded or unloaded, based on fluorescence spectroscopy and light scattering. In cytotoxicity tests, loaded and unloaded PACIS-b-PS-b-PEO nanoparticles proved nontoxic to various cell lines, including Balb/3T3 and MCF10A, with efficient cellular uptake, as shown by fluorescence imaging. Therefore, our findings highlight PACIS-b-PS-b-PEO as a versatile platform for preparing MCMs for multiple drug delivery.

Effects of Secondary Structures and pH on the Self-Assembly of Poly(ethylene glycol)-b-polytyrosine.

Different from conventional synthetic polymers, polypeptides exhibit a distinguishing characteristic of adopting specific secondary structures, including random coils, α-helixes, and β-sheets. The conformation determines the rigidity and solubility of polypeptide chains, which further direct the self-assembly and morphology of the nanostructures. We studied the effect of distinct secondary structures on the self-assembly behavior of polytyrosine (PTyr)-derived amphiphilic copolymers. Two block copolymers of enantiopure poly(ethylene glycol)-b-poly(l-tyrosine) (PEG-b-P(l-Tyr)) and racemic poly(ethylene glycol)-b-poly(dl-tyrosine) (PEG-b-P(dl-Tyr)) were synthesized through the ring-opening polymerization of l-tyrosine N-thiocarboxyanhydride (l-Tyr-NTA) and dl-tyrosine N-thiocarboxyanhydride (dl-Tyr-NTA), respectively, by using poly(ethylene glycol) amine as the initiator. PEG44-b-P(l-Tyr)10 adopts a β-sheet conformation and self-assembles into rectangular nanosheets in aqueous solutions, while PEG44-b-P(dl-Tyr)9 is primarily in a random coil conformation with a tiny content of β-sheet structures, which self-assembles into sheaf-like nanofibrils. A pH increase results in the ionization of phenolic hydroxyl groups, which decreases the β-sheet content and increases the random coil content of the PTyr segments. Accordingly, PEG44-b-P(l-Tyr)10 and PEG44-b-P(dl-Tyr)9 self-assemble to form slender nanobelts and twisted nanoribbons, respectively, in alkaline aqueous solutions. The secondary structure-driven self-assembly of PTyr-derived copolymers is promising to construct filamentous nanostructures, which have potential for applications in controlled drug release.

Synthesis and self-assembly of poly(4-vinylphenol)-b-poly(vinyl alcohol) diblock copolymer for invertible core-shell nanoparticles

We report the synthesis of the amphiphilic diblock copolymer, poly (vinyl phenol)-block-poly (vinyl alcohol), and its reversible formation of invertible core-shell nanoparticles. The diblock copolymer composed of poly (4-vinylphenol) (P4VPh) and poly (vinyl alcohol) (PVA) was prepared by the switchable reversible addition-fragmentation chain transfer polymerization (switchable-RAFT) of 4-acetoxystyrene and vinyl acetate and subsequent acetyl deprotection. The diblock copolymer was soluble in dimethyl sulfoxide (DMSO) but self-assembled into core-shell nanoparticles with the addition of H2O or tetrahydrofuran (THF). The formation and transition of the invertible core-shell system in response to solvent composition was characterized by 1H NMR, dynamic light scattering, and transmission electron microscopy. Interestingly, the fluorescent behavior of the P4VPh block was changed according to the morphological change of core-shell nanoparticle induced by solvent composition. With increasing the H2O content, the P4VPh chains in the DMSO solution collapsed and the diblock copolymers self-assembled to core-shell type micelles. Aggregation of P4VPh chains increased the local concentration of the phenolic group, resulting in fluorescence quenching. However, with the addition of THF to the DMSO solution, the inverted core-shell nanoparticles were formed where segregated P4VPh chains showed fluorescence.

Protocells Featuring Membrane-Bound and Dynamic Membraneless Organelles.

Living cells, especially eukaryotic ones, use multicompartmentalization to regulate intra- and extracellular activities, featuring membrane-bound and membraneless organelles. These structures govern numerous biological and chemical processes spatially and temporally. Synthetic cell models, primarily utilizing lipidic and polymeric vesicles, have been developed to carry out cascade reactions within their compartments. However, these reconstructions often segregate membrane-bound and membraneless organelles, neglecting their collaborative role in cellular regulation. To address this, we propose a structural design incorporating microfluidic-produced liposomes housing synthetic membrane-bound organelles made from self-assembled poly(ethylene glycol)-block-poly(trimethylene carbonate) nanovesicles and synthetic membraneless organelles formed via temperature-sensitive elastin-like polypeptide phase separation. This architecture mirrors natural cellular organization, facilitating a detailed examination of the interactions for a comprehensive understanding of cellular dynamics.

Highly Efficient Fabrication of Fluorescent "Turn-On" Lateral Flow Strips for Highly Sensitive Detection of Small Molecules Based on Self-Assembly of AuAg Nanoclusters.

Currently, fluorescent "turn-on" lateral flow assay (FONLFA) has shown enhanced "naked eye" detection sensitivity for small molecules, while it is urgent to adopt biocompatible fluorescent nanomaterials and needs new strategies to simplify the preparation process. In this study, a highly effective method was proposed to produce FONLFA strips for the detection of small molecules. The gold-silver nanoclusters (AuAgNCs) were immobilized onto the nitrocellulose membrane of the strips by the self-assembly of poly(sodium 4-styrenesulfonate), antigen, and AuAgNCs. The immobilization process entails a straightforward mixing of the three components, taking merely 1 min, thereby bypassing the necessity for chemical modification of fluorescent nanomaterials. The strategy offers a significantly simplified process, which substantially enhances the efficiency of the strip fabrication. Utilizing this method, a FONLFA was developed for carbendazim with a visual limit of detection (vLOD) reduced by 40-fold compared with the conventional colorimetric lateral flow assay (LFA). Furthermore, the approach demonstrates versatility by enabling the immobilization of AuAgNCs and streptavidin, which facilitates the development of aptamer-based FONLFAs. The designed aptamer-based FONLFA for kanamycin exhibited a 50-fold reduction in the vLOD compared with conventional colorimetric LFAs. Therefore, FONLFA holds promising potential for widespread applications in the analysis of small molecules.

A Versatile Synthesis Platform Based on Polymer Cubosomes for a Library of Highly Ordered Nanoporous Metal Oxides Particles.

Polymer cubosomes (PCs) have well-defined inverse bicontinuous cubic mesophases formed by amphiphilic block copolymer bilayers. The open hydrophilic channels, large periods, and robust physical properties of PCs are advantageous to many host-guest interactions and yet not fully exploited, especially in the fields of functional nanomaterials. Here, the self-assembly of poly(ethylene oxide)-block-polystyrene block copolymers is systematically investigated and a series of robust PCs is developed via a cosolvent method. Ordered nanoporous metal oxide particles are obtained by selectively filling the hydrophilic channels of PCs via an impregnation strategy, followed by a two-step thermal treatment. Based on this versatile PC platform, the general synthesis of a library of ordered porous particles with different pore structures , tunable large pore size (18-78nm), high specific surface areas (up to 123.3m2g-1 for WO3) and diverse framework compositions, such as transition and non-transition metal oxides, rare earth chloride oxides, perovskite, pyrochlore, and high-entropy metal oxides is demonstrated. As typical materials obtained via this method, ordered porous WO3 particles have the advantages of open continuous structure and semiconducting properties, thus showing superior gas sensing performances toward hydrogen sulfide.

Plasmonic Properties of Self-Assembled Gold Nanocrescents: Implications for Chemical Sensing.

A bottom-up approach, the Langmuir-Blodgett technique, is used for the preparation of composite thin films of gold nanoparticles and polymers: poly(styrene-b-2-vinylpyridine), poly-2-vinylpyridine, and polystyrene. The self-assembly of poly(styrene-b-2-vinylpyridine) at the air-water interface leads to the formation of surface micelles, which serve as a template for the organization of gold nanoparticles into ring assemblies. By using poly-2-vinylpyridine in conjunction with low surface pressure, the distance between nanostructures can be increased, allowing for optical characterization of single nanostructures. Once deposited on a solid substrate, the preorganized gold nanoparticles are subjected to further growth by the reduction of additional gold, leading to a variety of nanostructures which can be divided into two categories: nanocrescents and circular arrays of nanoparticles. The optical properties of individual structures are investigated by optical dark-field spectroscopy and numerical calculations. The plasmonic behavior of the nanostructures is elucidated through the correlation of optical properties with structural features and the identification of dominant plasmon modes. Being based on a self-assembly approach, the reported method allows for the formation of interesting plasmonic materials under ambient conditions, at a relatively large scale, and at low cost. These attributes, in addition to the resonances located in the near-infrared region of the spectrum, make nanocrescents candidates for biological and chemical sensing.

Synthesis via Reversible Addition–Fragmentation Chain Transfer Polymerization and Deep Control Property of Amphiphilic Self-assembled Poly(ethylene oxide)-block-polystyrene

Synthesis via Reversible Addition–Fragmentation Chain Transfer Polymerization and Deep Control Property of Amphiphilic Self-assembled Poly(ethylene oxide)-<i>block</i>-polystyrene

Controlled adsorption of multiple bioactive proteins enables targeted mast cell nanotherapy.

Protein adsorption onto nanomaterials often results in denaturation and loss of bioactivity. Controlling the adsorption process to maintain the protein structure and function has potential for a range of applications. Here we report that self-assembled poly(propylene sulfone) (PPSU) nanoparticles support the controlled formation of multicomponent enzyme and antibody coatings and maintain their bioactivity. Simulations indicate that hydrophobic patches on protein surfaces induce a site-specific dipole relaxation of PPSU assemblies to non-covalently anchor the proteins without disrupting the protein hydrogen bonding or structure. As a proof of concept, a nanotherapy employing multiple mast-cell-targeted antibodies for preventing anaphylaxis is demonstrated in a humanized mouse model. PPSU nanoparticles displaying an optimized ratio of co-adsorbed anti-Siglec-6 and anti-FcεRIα antibodies effectively inhibit mast cell activation and degranulation, preventing anaphylaxis. Protein immobilization on PPSU surfaces provides a simple and rapid platform for the development of targeted protein nanomedicines.

A Spike-like Self-Assembly of Polyaspartamide Integrated with Functionalized Nanoparticles.

The integration of nanoparticles (NPs) into molecular self-assemblies has been extensively studied with the aim of building well-defined, ordered structures which exhibit advanced properties and performances. This study demonstrates a novel strategy for the preparation of a spike-like self-assembly designed to enhance UV blocking. Poly(2-hydroxyethyl aspartamide) (PHEA) substituted with octadecyl chains and menthyl anthranilate (C18-M-PHEA) was successfully synthesized by varying the number of grafted groups to control their morphology and UV absorption. The in situ incorporation of polymerized rod-like TiO2 within the C18-M-PHEA self-aggregates generated spike-like self-assemblies (TiO2@C18-M-PHEA) with a chestnut burr structure in aqueous solution. The results showed that the spike-like self-assemblies integrated with TiO2 NPs exhibited a nine-fold increase in UV protection by simultaneous UV absorption and scattering compared with the pure TiO2 NPs formed via a bulk mixing process. This work provides a novel method for UV protection using self-assembling poly(amino acid)s derivatives integrated with functional nanoparticles to tune their morphology and organization.

Nanoparticle-Conjugated Toll-Like Receptor 9 Agonists Improve the Potency, Durability, and Breadth of COVID-19 Vaccines.

Development of effective vaccines for infectious diseases has been one of the most successful global health interventions in history. Though, while ideal subunit vaccines strongly rely on antigen and adjuvant(s) selection, the mode and time scale of exposure to the immune system has often been overlooked. Unfortunately, poor control over the delivery of many adjuvants, which play a key role in enhancing the quality and potency of immune responses, can limit their efficacy and cause off-target toxicities. There is a critical need for improved adjuvant delivery technologies to enhance their efficacy and boost vaccine performance. Nanoparticles have been shown to be ideal carriers for improving antigen delivery due to their shape and size, which mimic viral structures but have been generally less explored for adjuvant delivery. Here, we describe the design of self-assembled poly(ethylene glycol)-b-poly(lactic acid) nanoparticles decorated with CpG, a potent TLR9 agonist, to increase adjuvanticity in COVID-19 vaccines. By controlling the surface density of CpG, we show that intermediate valency is a key factor for TLR9 activation of immune cells. When delivered with the SARS-CoV-2 spike protein, CpG nanoparticle (CpG-NP) adjuvant greatly improves the magnitude and duration of antibody responses when compared to soluble CpG, and results in overall greater breadth of immunity against variants of concern. Moreover, encapsulation of CpG-NP into injectable polymeric-nanoparticle (PNP) hydrogels enhances the spatiotemporal control over codelivery of CpG-NP adjuvant and spike protein antigen such that a single immunization of hydrogel-based vaccines generates humoral responses comparable to those of a typical prime-boost regimen of soluble vaccines. These delivery technologies can potentially reduce the costs and burden of clinical vaccination, both of which are key elements in fighting a pandemic.

Self-assembly of poly(ionic liquid) block copolymer based dielectrics on semiconductor formation and performance.

Self-assembly of poly(ionic liquid) block copolymer based dielectrics on semiconductor formation and performance.

Self-assembled poly[2]catenanes based on non-covalent and dynamic covalent bonds

We report totally self-assembled poly[2]catenanes from M1 and M2 based on non-covalent and dynamic covalent bonds. M1 first self-assembled into SPs. Then, the addition of M2 induced dynamic covalent assembly with SPs to form poly[2]catenanes.

Combined ROS Sensitive PEG-PPS-PEG with Peptide Agonist for Effective Target Therapy in Mouse Model.

Growth hormone-releasing hormone (GHRH) agonist, a 29-amino acid peptide, shows significant potential in treating myocardial infarction (MI) by aiding the repair of injured heart tissue. The challenge lies in the effective on-site delivery of GHRH agonist. This study explores the use of a targetable delivery system employing ROS-responsive PEG-PPS-PEG polymers to encapsulate and deliver GHRH agonist MR409 for enhanced therapeutic efficacy. We synthesized a self-assembling poly (ethylene glycol)-poly (propylene sulfide)-poly (ethylene glycol) polymer (PEG-PPS-PEG) amphiphilic polymer responsive to reactive oxygen species (ROS). The hydrophilic peptide GHRH agonist MR409 was encapsulated within these polymers to form nano PEG-PPS-PEG@MR409 vesicles (NPs). Cardiomyocyte apoptosis was induced under hypoxia and serum-free culture condition for 24 hours, and their production of ROS was detected by fluorescence dye staining. The cellular uptake of PEG-PPS-PEG@MR409 NPs was observed using fluorescence-labeled MR409. Targeting ability and therapeutic efficacy were evaluated using a mouse MI model. PEG-PPS-PEG@MR409 NPs were efficiently internalized by cardiomyocytes, reducing ROS levels and apoptosis. These NPs exhibited superior targeting to the infarcted heart compared to naked MR409 peptide. With a reduced injection frequency (once every three days), PEG-PPS-PEG@MR409 NPs significantly promoted cardiac function recovery post-MI, matching the efficacy of daily MR409 injections. ROS-responsive PEG-PPS-PEG polymers provide a novel and effective platform for the targeted delivery of GHRH agonist peptides, improving cardiac function and offering a new approach for peptide therapy in MI treatment.

The effect of comb length on the in vitro and in vivo properties of self-assembled poly(oligoethylene glycol methacrylate)-based block copolymer nanoparticles.

Comb copolymer analogues of poly(lactic acid)-polyethylene glycol block copolymers (PLA-b-PEG) offer potential to overcome the inherent chemistry and stability limitations of their linear block copolymer counterparts. Herein, we examine the differences between P(L)LA10K-b-PEG10K and linear-comb copolymer analogues thereof in which the linear PEG block is replaced by poly(oligo(ethylene glycol) methacrylate) (POEGMA) blocks with different side chain (comb) lengths but the same overall molecular weight. P(L)LA10K-b-POEGMA47510K and P(L)LA10K-b-POEGMA200010K block copolymers were synthesized via activators regenerated by electron transfer atom transfer radical polymerization (ARGET ATRP) and fabricated into self-assembled nanoparticles using flash nanoprecipitation via confined impinging jet mixing. Linear-comb copolymer analogues based on PLA-b-POEGMA yielded smaller but still well-controlled nanoparticle sizes (88 ± 2 nm and 114 ± 1 nm respectively compared to 159 ± 2 nm for P(L)LA10K-b-PEG10K nanoparticles) that exhibited improved colloidal stability relative to linear copolymer-based nanoparticles over a 15 day incubation period while maintaining comparably high cytocompatibility, although the comb copolymer analogues had somewhat lower loading capacity for doxorubicin hydrochloride. Cell spheroid studies showed that the linear-comb copolymers promoted enhanced tumor transport and thus cell killing compared to conventional linear block copolymers. In vivo studies showed all NP types could passively accumulate within implanted CT26 tumors but with different accumulation profiles, with P(L)LA10K-b-POEGMA200010K NPs showing continuous accumulation throughout the full 24 h monitoring period whereas tumor accumulation of P(L)LA10K-b-POEGMA47510K NPs was significant only between 8 h and 24 h. Overall, the linear-comb copolymer analogues exhibited superior stability, biodistribution, spheroid penetration, and inherent tunability over linear NP counterparts.

Self-assembled poly(lactic acid) films with high heat resistance and multi-stage thermochromic properties prepared by blown film-annealing

Developing reversible thermochromic films that can achieve thermochromism from low to high temperatures by simple processes is a goal which carries a great deal of potential. They have potential applications in temperature monitoring, information encryption and anti-counterfeiting. For this work, decanedioic acid, 1,10-bis(2-benzoylhydrazide) (TMC-300) was used as a self-assembled organic nucleating agent and was combined with an annealing process to heat-resistant modify the blown polylactic acid films. The thermochromic films would have a higher upper limit to its detection range. Through this method, we obtained thermochromic films with a dense morphological structure. Additionally, the effects of TMC-300 and annealing on various properties of films were studied in detail. Unexpectedly, we found a hindering effect between thermochromic microcapsule (TCM) and TMC-300 at TMC-300 mass fractions below 0.3 wt%. Subsequently, TCM with different color changes in different temperature ranges was added. Films with thermochromic color change in the range of 25–70 °C were successfully prepared by this method. The color of the films also demonstrated a multi-stage thermochromic transition through brown, orange, and red before eventually becoming colorless. The results showed the crystallinity of the films rose to 45.65 %, while the heat resistance temperature and elongation at break increased by 60 % and 170 %, respectively. The prepared thermochromic films had enough transparency and even after 60 heating–cooling cycles, they exhibited outstanding thermochromic properties. Moreover, we have explored the application of films in fields of information encryption and temperature monitoring by utilising their multi-stage thermochromic properties.

Yolk-Shell Encapsulation of Cells by Biomimetic Mineralization and Visible Light-Induced Surface Graft Polymerization.

The pursuit of low-cytotoxicity modification strategies represents a prominent avenue in cell coating research, holding immense significance for the advancement of practical living cell-related technologies. Here, we presented a novel method to fabricate encapsulated yeast cells with a yolk-shell structure by biomimetic mineralization and visible-light-induced surface graft polymerization. In this approach, an amorphous calcium carbonate (ACC) shell was first deposited on the surface of a yeast cell (cell@ACC) modified with 4 layers of self-assembled poly(diallyl dimethylammonium chloride) (PDADMAC)/poly(acrylic acid) (PAA) film using a biomimetic mineralization technique. Subsequently, polyethylenimine (PEI) was absorbed on the surface of cell@ACC by electrostatic interaction. Then, a cross-linked shell was introduced by surface-initiated graft polymerization of poly(ethylene glycol) diacrylate (PEGDA) on cell@ACC under irradiation of visible light using thioxanthone catechol-O,O'-diacetic acid as the photosensitizer. After the removal of the inner ACC shell, the yolk-shell-structured yeast cells (cell@PHS) were obtained. Due to the mild conditions of the strategy, the cell@PHS could retain 98.81% of its original viability. The introduction of the shell layer significantly prolonged the lag phase of yeast cells, which could be tuned between 5 and 25 h by regulating the thickness of the shell. Moreover, the cell@PHS showed improved resistance against lyticase due to the presence of a protective shell. After 30 days of storage, the viability of cell@PHS was 81.09%, which is significantly higher than the 19.89% viability of native yeast cells.