Nanoarchitectonics of Bactericidal Coatings Based on CaCO3-Nanosilver Hybrids.

Antimicrobial coatings provide protection against microbes colonization on surfaces. This can prevent the stabilization and proliferation of microorganisms. The ever-increasing levels of microbial resistance to antimicrobials are urging the development of alternative types of compounds that are potent across broad spectra of microorganisms and target different pathways. This will help to slow down the development of resistance and ideally halt it. The development of composite antimicrobial coatings (CACs) that can host and protect various antimicrobial agents and release them on demand is an approach to address this urgent need. In this work, new CACs based on microsized hybrids of calcium carbonate (CaCO3) and silver nanoparticles (AgNPs) were designed using a drop-casting technique. Polyvinylpyrrolidone and mucin were used as additives. The CaCO3/AgNPs hybrids contributed to endowing colloidal stability to the AgNPs and controlling their release, thereby ensuring the antibacterial activity of the coatings. Moreover, the additives PVP and mucin served as a matrix to (i) control the distribution of the hybrids, (ii) ensure mechanical integrity, and (iii) prevent the undesired release of AgNPs. Scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FTIR) techniques were used to characterize the 15 µm thick CAC. The antibacterial activity was determined against Escherichia coli, methicillin-resistant Staphylococcus aureus (MRSA), and Pseudomonas aeruginosa, three bacteria responsible for many healthcare infections. Antibacterial performance of the hybrids was demonstrated at concentrations between 15 and 30 µg/cm2. Unloaded CaCO3 also presented bactericidal properties against MRSA. In vitro cytotoxicity tests demonstrated that the hybrids at bactericidal concentrations did not affect human dermal fibroblasts and human mesenchymal stem cell viability. In conclusion, this work presents a simple approach for the design and testing of advanced multicomponent and functional antimicrobial coatings that can protect active agents and release them on demand.

Comparative Characterization of Iron and Silver Nanoparticles: Extract-Stabilized and Classical Synthesis Methods.

Synthesis of silver nanoparticles using extracts from plants is an advantageous technological alternative to the traditional colloidal synthesis due to its simplicity, low cost, and the inclusion of environmentally friendly processes to obtain a new generation of antimicrobial compounds. The work describes the production of silver and iron nanoparticles using sphagnum extract as well as traditional synthesis. Dynamic light scattering (DLS) and laser doppler velocimetry methods, UV-visible spectroscopy, transmission electron microscopy (TEM) combined with energy dispersive X-ray spectroscopy (EDS), atomic force microscopy (AFM), dark-field hyperspectral microscopy, and Fourier-transform infrared spectroscopy (FT-IR) were used to study the structure and properties of synthesized nanoparticles. Our studies demonstrated a high antibacterial activity of the obtained nanoparticles, including the formation of biofilms. Nanoparticles synthesized using sphagnum moss extracts likely have high potential for further research.

How similar is the antibacterial activity of silver nanoparticles coated with different capping agents?

Silver nanoparticles (AgNPs) represent one of the most commercialised metal nanomaterials, with an extensive number of applications that span from antimicrobial products to electronics. Bare AgNPs are very susceptible to aggregation, and capping agents are required for their protection and stabilisation. The capping agents can endow new characteristics which can either improve or deteriorate AgNPs (bio)activity. In the present work, five different capping agents were studied as stabilizing agents for AgNPs: trisodium citrate (citrate), polyvinylpyrrolidone (PVP), dextran (Dex), diethylaminoethyl-dextran (DexDEAE) and carboxymethyl-dextran (DexCM). The properties of the AgNPs were studied using a set of methods, including transmission electron microscopy, X-ray diffraction, thermogravimetric analysis and ultraviolet-visible and infrared spectroscopy. Coated and bare AgNPs were also tested against Escherichia coli, methicillin-resistance Staphylococcus aureus and Pseudomonas aeruginosa to analyse their capacity to suppress bacterial growth and eradicate biofilms of clinically relevant bacteria. The results showed that all the capping agents endow long-term stability for the AgNPs in water; however, when the AgNPs are in bacterial culture media, their stability is highly dependent on the capping agent properties due to the presence of electrolytes and charged macromolecules such as proteins. The results also showed that the capping agents have a substantial impact on the antibacterial activity of the AgNPs. The AgNPs coated with the Dex and DexCM were the most effective against the three strains, due to their better stability which resulted in the release of more silver ions, better interactions with the bacteria and diffusion into the biofilms. It is hypothesized that the antibacterial activity of capped AgNPs is governed by a balance between the AgNPs stability and their ability to release silver ions. Strong adsorption of capping agents like PVP on the AgNPs endows higher colloidal stability in culture media; however, it can decrease the rate of Ag+ release from the AgNPs and reduce the antibacterial performance. Overall, this work presents a comparative study between different capping agents on the properties and antibacterial activity of AgNPs, highlighting the importance of the capping agent in their stability and bioactivity.

Interaction of Lysozyme with Poly(L-lysine)/Hyaluronic Acid Multilayers: An ATR-FTIR Study.

Polyelectrolyte multilayers (PEM) loaded with bioactive molecules such as proteins serve as excellent mimics of an extracellular matrix and may find applications in fields such as biomedicine and cell biology. A question which is crucial to the successful employment of PEMs is whether conformation and bioactivity of the loaded proteins is preserved. In this work, the polarized attenuated total reflection Fourier transform infrared (ATR-FTIR) technique is applied to investigate the conformation of the protein lysozyme (Lys) loaded into the poly(L-lysine)/hyaluronic acid (PLL/HA) multilayers. Spectra are taken from the protein in the PEMs coated onto an ATR crystal during protein adsorption and desorption. For comparison, a similar investigation is performed for the case of Lys in contact with the uncoated crystal. The study highlights the presence of both "tightly" and "poorly bound" Lys fractions in the PEM. These fractions differ in their conformation and release behavior from the PEM upon washing. Comparison of spectra recorded with different polarizations suggests preferential orientation of alpha helical structures, beta sheets and turns in the "tightly bound" Lys. In contrast, the "poorly bound" fraction shows isotropic orientation and its conformation is well preserved.

A lipid membrane supported on an artificial extracellular matrix made of polyelectrolyte multilayers: towards nanoarchitectonics at the cellular interface.

To implement a specific function, cells recognize multiple physical and chemical cues and exhibit molecular responses at their interfaces - the boundary regions between the cell lipid-based membrane and the surrounding extracellular matrix (ECM). Mimicking the cellular external microenvironment presents a big challenge in nanoarchitectonics due to the complexity of the ECM and lipid membrane fragility. This study reports an approach for the assembly of a lipid bilayer, mimicking the cellular membrane, placed on top of a polyelectrolyte multilayer cushion made of hyaluronic acid and poly-L-lysine - a nanostructured biomaterial, which represents a 3D artificial ECM. Model proteins, lysozyme and α-lactalbumin, (which have similar molecular masses but carry opposite net charges) have been employed as soluble signalling molecules to probe their interaction with these hybrids. The formation of a lipid bilayer and the intermolecular interactions in the hybrid structure are monitored using a quartz crystal microbalance and confocal fluorescence microscopy. Electrostatic interactions between poly-L-lysine and the externally added proteins govern the transport of proteins into the hybrid. Designed ECM-cell mimicking hybrids open up new avenues for modelling a broad range of cell membranes and ECM and their associated phenomena, which can be used as a tool for synthetic biology and drug screening.

Spontaneous shrinkage drives macromolecule encapsulation into layer-by-layer assembled biopolymer microgels.

HYPOTHESIS: Recently, the anomalous shrinkage of surface-supported hyaluronate/poly-l-lysine (HA/PLL) microgels (µ-gels), which exceeds that reported for any other multilayer-based systems, has been reported [1]. The current study investigates the capability of these unique µ-gels for the encapsulation and retention of macromolecules, and proposes the shrinkage-driven assembly of biopolymer-based µ-gels as a novel tool for one-step surface biofunctionalization. EXPERIMENTS: A set of dextrans (DEX) and their charged derivatives - carboxymethyl (CM)-DEX and diethylaminoethyl (DEAE)-DEX - has been utilized to evaluate the effects of macromolecular mass and net charge on µ-gel shrinkage and macromolecule entrapment. µ-gels formation on the surface of silicone catheters exemplifies their potential to tailor biointerfaces. FINDINGS: Shrinkage-driven µ-gel formation strongly depends on the net charge and mass content of encapsulated macromolecules. Inclusion of neutral DEX decreases the degree of shrinkage several times, whilst charged DEXs adopt to the backbone of oppositely charged polyelectrolytes, resulting in shrinkage comparable to that of non-loaded µ-gels. Retention of CM-DEX in µ-gels is significantly higher compared to DEAE-DEX. These insights into the mechanisms of macromolecular entrapment into biopolymer-based µ-gels promotes fundamental understanding of molecular dynamics within the multilayer assemblies. Organization of biodegradable µ-gels at biomaterial surfaces opens avenues for their further exploitation in a diverse array of bioapplications.

Vaterite vectors for the protection, storage and release of silver nanoparticles.

Silver nanoparticles (AgNPs) have found widespread commercial applications due to their unique physical and chemical properties. However, their relatively poor stability remains a main problem. An ideal way to improve the stability of AgNPs is not only to endow colloidal stability to individual nanoparticles but also to protect them from environmental factors that induce their agglomeration, like variation of ionic strength and pH, presence of macromolecules, etc. Mesoporous calcium carbonate vaterite crystals (CaCO3 vaterite) have recently attracted significant attention as inexpensive and biocompatible carriers for the encapsulation and controlled release of both drugs and nanoparticles. This work aimed to develop an approach to load AgNPs into CaCO3 vaterite without affecting their properties. We focused on improving the colloidal stability of AgNPs by using different capping agents, and understanding the mechanism behind AgNPs loading and release from CaCO3 crystals. Various methods were applied to study the AgNPs and CaCO3 crystals loaded with AgNPs (CaCO3/AgNPs hybrids), such as scanning and transmission electron microscopy, X-ray diffraction, infrared and mass spectrometry. The results demonstrated that polyvinylpyrrolidone and positively charged diethylaminoethyl-dextran can effectively keep the colloidal stability of AgNPs during co-precipitation with CaCO3 crystals. CaCO3/AgNPs hybrids composed of up to 4 % weight content of nanoparticles were produced, with the loading mechanism being well-described by the Langmuir adsorption model. In vitro release studies demonstrated a burst release of stable AgNPs at pH 5.0 and a sustained release at pH 7.5 and 9.0. The antibacterial studies showed that these hybrids are effective against Escherichia coli, methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa, three important bacteria responsible for nosocomial infections. The developed approach opens a new way to stabilise, protect, store and release AgNPs in a controlled manner for their use as antimicrobial agents.

Formulation and Biodegradation of Surface-Supported Biopolymer-Based Microgels Formed via Hard Templating onto Vaterite CaCO3 Crystals.

In recent decades, there has been increased attention to the role of layer-by-layer assembled bio-polymer 3D structures (capsules, beads, and microgels) for biomedical applications. Such free-standing multilayer structures are formed via hard templating onto sacrificial cores such as vaterite CaCO3 crystals. Immobilization of these structures onto solid surfaces (e.g., implants and catheters) opens the way for the formulation of advanced bio-coating with a patterned surface. However, the immobilization step is challenging. Multiple approaches based mainly on covalent binding have been developed to localize these multilayer 3D structures at the surface. This work reports a novel strategy to formulate multilayer surface-supported microgels (ss-MG) directly on the surface via hard templating onto ss-CaCO3 pre-grown onto the surface via the direct mixing of Na2CO3 and CaCl2 precursor solutions. ss-MGs were fabricated using biopolymers: polylysine (PLL) as polycation and three polyanions-hyaluronic acid (HA), heparin sulfate (HS), and alginate (ALG). ss-MG biodegradation was examined by employing the enzyme trypsin. Our studies indicate that the adhesion of the ss-MG to the surface and its formation yield directly correlate with the mobility of biopolymers in the ss-MG, which decreases in the sequence of ALG > HA > HS-based ss-MGs. The adhesion of HS-based ss-MGs is only possible via heating during their formation. Dextran-loading increases ss-MG formation yield while reducing ss-MG shrinking. ss-MGs with higher polymer mobility possess slower biodegradation rates, which is likely due to diffusion limitations for the enzyme in more compact annealed ss-MGs. These findings provide valuable insights into the mechanisms underlying the formation and biodegradation of surface-supported biopolymer structures.

Surface Modification with Particles Coated or Made of Polymer Multilayers.

The coating of particles or decomposable cores with polyelectrolytes via Layer-by-Layer (LbL) assembly creates free-standing LbL-coated functional particles. Due to the numerous functions that their polymers can bestow, the particles are preferentially selected for a plethora of applications, including, but not limited to coatings, cargo-carriers, drug delivery vehicles and fabric enhancements. The number of publications discussing the fabrication and usage of LbL-assembled particles has consistently increased over the last vicennial. However, past literature fails to either mention or expand upon how these LbL-assembled particles immobilize on to a solid surface. This review evaluates examples of LbL-assembled particles that have been immobilized on to solid surfaces. To aid in the formulation of a mechanism for immobilization, this review examines which forces and factors influence immobilization, and how the latter can be confirmed. The predominant forces in the immobilization of the particles studied here are the Coulombic, capillary, and adhesive forces; hydrogen bonding as well as van der Waal's and hydrophobic interactions are also considered. These are heavily dependent on the factors that influenced immobilization, such as the particle morphology and surface charge. The shape of the LbL particle is related to the particle core, whereas the charge was dependant on the outermost polyelectrolyte in the multilayer coating. The polyelectrolytes also determine the type of bonding that a particle can form with a solid surface. These can be via either physical (non-covalent) or chemical (covalent) bonds; the latter enforcing a stronger immobilization. This review proposes a fundamental theory for immobilization pathways and can be used to support future research in the field of surface patterning and for the general modification of solid surfaces with polymer-based nano- and micro-sized polymer structures.

Hybrid Mucin-Vaterite Microspheres for Delivery of Proteolytic Enzyme Chymotrypsin.

While the enteral delivery of proteolytic enzymes is widely established for combating many diseases as an alternative to antibiotic treatment, their local delivery only emerges as administration route enabling sustained release in a controlled manner on site. The latest requires the development of drug delivery systems suitable for encapsulation and preservation of enzymatic proteolytic activity. This study proposes hybrid microspheres made of mucin and biodegradable porous crystals of calcium carbonate (CC) as the carriers for chymotrypsin (CTR) delivery. CTR is impregnated into CC and hybrid CC/mucin (CCM) microspheres by means of sorption without any chemical modification. The loading of the CC with mucin enhances CTR retention on hybrid microspheres (adsorption capacity of ≈8.7 mg g-1  vs 4.7 mg g-1 ), recharging crystal surface due to the presence of mucin and diminishing the average pore diameter of the crystals from 25 to 8 nm. Mucin also retards recrystallization of vaterite into nonporous calcite improving stability of CCM microspheres upon storage. Proteolytic activity of CTR is preserved in both CC and CCM microspheres, being pH dependent. Temperature-induced inactivation of CTR significantly diminishes by CTR encapsulation into CC and CCM microspheres. Altogether, these findings indicate promises of hybrid mucin-vaterite microspheres for mucosal application of proteases.

Modification of Surfaces with Vaterite CaCO3 Particles.

Former studies have demonstrated a strong interest toward the crystallization of CaCO3 polymorphs in solution. Nowadays, CaCO3 crystallization on solid surfaces is extensively being studied using biomolecules as substrates for the control of the growth aiming at various applications of CaCO3. Calcium carbonate exists in an amorphous state, as three anhydrous polymorphs (aragonite, calcite and vaterite), and as two hydrated polymorphs (monohydrocalcite and ikaite). The vaterite polymorph is considered as one of the most attractive forms due to its large surface area, biocompatibility, mesoporous nature, and other features. Based on physical or chemical immobilization approaches, vaterite can be grown directly on solid surfaces using various (bio)molecules, including synthetic polymers, biomacromolecules such as proteins and peptides, carbohydrates, fibers, extracellular matrix components, and even biological cells such as bacteria. Herein, the progress on the modification of solid surfaces by vaterite CaCO3 crystals is reviewed, focusing on main findings and the mechanism of vaterite growth initiated by various substances mentioned above, as well as the discussion of the applications of such modified surfaces.

The influence of Ca/Mg ratio on autogelation of hydrogel biomaterials with bioceramic compounds.

Hydrogels, which are versatile three-dimensional structures containing polymers and water, are very attractive for use in biomedical fields, but they suffer from rather weak mechanical properties. In this regard, biocompatible particles can be used to enhance their mechanical properties. The possibility of loading such particles with drugs (e.g. enzymes) makes them a particularly useful component in hydrogels. In this study, micro/nanoparticles containing various ratios of Ca2+/Mg2+ with sizes ranging from 1 to 8 µm were prepared and mixed with gellan gum (GG) solution to study the in-situ formation of hydrogel-particle composites. The particles provide multiple functionalities: 1) they efficiently crosslink GG to induce hydrogel formation through the release of the divalent cations (Ca2+/Mg2+) known to bind to GG polymer chains; 2) they enhance mechanical properties of the hydrogel from 2 up to 100 kPa; 3) the samples most efficiently promoting cell growth were found to contain two types of minerals: vaterite and hydroxymagnesite, which enhanced cells proliferation and hydroxyapatite formation. The results demonstrate that such composite materials are attractive candidates for applications in bone regeneration.

Mesoporous One-Component Gold Microshells as 3D SERS Substrates.

Surface-enhanced Raman scattering (SERS) is a powerful analytical tool for label-free analysis that has found a broad spectrum of applications in material, chemical, and biomedical sciences. In recent years, a great interest has been witnessed in the rational design of SERS substrates to amplify Raman signals and optionally allow for the selective detection of analytes, which is especially essential and challenging for biomedical applications. In this study, hard templating of noble metals is proposed as a novel approach for the design of one-component tailor-made SERS platforms. Porous Au microparticles were fabricated via dual ex situ adsorption of Au nanoparticles and in situ reduction of HAuCl4 on mesoporous sacrificial microcrystals of vaterite CaCO3. Elimination of the microcrystals at mild conditions resulted in the formation of stable mesoporous one-component Au microshells. SERS performance of the microshells at very low 0.4 µW laser power was probed using rhodamine B and bovine serum albumin showing enhancement factors of 2 × 108 and 8 × 108, respectively. The proposed strategy opens broad avenues for the design and scalable fabrication of one-component porous metal particles that can serve as superior SERS platforms possessing both excellent plasmonic properties and the possibility of selective inclusion of analyte molecules and/or SERS nanotags for highly specific SERS analysis.

Pharmaceuticals Removal by Adsorption with Montmorillonite Nanoclay.

The problem of purifying domestic and hospital wastewater from pharmaceutical compounds is becoming more and more urgent every year, because of the continuous accumulation of chemical pollutants in the environment and the limited availability of freshwater resources. Clay adsorbents have been repeatedly proposed as adsorbents for treatment purposes, but natural clays are hydrophilic and can be inefficient for catching hydrophobic pharmaceuticals. In this paper, a comparison of adsorption properties of pristine montmorillonite (MMT) and montmorillonite modified with stearyl trimethyl ammonium (hydrophobic MMT-STA) towards carbamazepine, ibuprofen, and paracetamol pharmaceuticals was performed. The efficiency of adsorption was investigated under varying solution pH, temperature, contact time, initial concentration of pharmaceuticals, and adsorbate/adsorbent mass ratio. MMT-STA was better than pristine MMT at removing all the pharmaceuticals studied. The adsorption capacity of hydrophobic montmorillonite to pharmaceuticals decreased in the following order: carbamazepine (97%) > ibuprofen (95%) > paracetamol (63-67%). Adsorption isotherms were best described by Freundlich model. Within the pharmaceutical concentration range of 10-50 µg/mL, the most optimal mass ratio of adsorbates to adsorbents was 1:300, pH 6, and a temperature of 25 °C. Thus, MMT-STA could be used as an efficient adsorbent for deconta×ating water of carbamazepine, ibuprofen, and paracetamol.

Which Biopolymers Are Better for the Fabrication of Multilayer Capsules? A Comparative Study Using Vaterite CaCO3 as Templates.

The polymer layer-by-layer assembly is accounted among the most attractive approaches for the design of advanced drug delivery platforms and biomimetic materials in 2D and 3D. The multilayer capsules can be made of synthetic or biologically relevant (e.g., natural) polymers. The biopolymers are advantageous for bioapplications; however, the design of such "biocapsules" is more challengeable due to intrinsic complexity and lability of biopolymers. Until now, there are no systematic studies that report the formation mechanism for multilayer biocapsules templated upon CaCO3 crystals. This work evaluates the structure-property relationship for 16 types of capsules made of different biopolymers and proposes the capsule formation mechanism. The capsules have been fabricated upon mesoporous cores of vaterite CaCO3, which served as a sacrificial template. Stable capsules of polycations poly-l-lysine or protamine and four different polyanions were successfully formed. However, capsules made using the polycation collagen and dextran amine underwent dissolution. Formation of the capsules has been correlated with the stability of the respective polyelectrolyte complexes at increased ionic strength. All formed capsules shrink upon core dissolution and the degree of shrinkage increased in the series of polyanions: heparin sulfate < dextran sulfate < chondroitin sulfate < hyaluronic acid. The same trend is observed for capsule adhesiveness to the glass surface, which correlates with the decrease in polymer charge density. The biopolymer length and charge density govern the capsule stability and internal structure; all formed biocapsules are of a matrix-type, other words are microgels. These findings can be translated to other biopolymers to predict biocapsule properties.

Cooling-Triggered Release from Mesoporous Poly(N-isopropylacrylamide) Microgels at Physiological Conditions.

Poly(N-isopropylacrylamide) (pNIPAM) hydrogels have broad potential applications as drug delivery vehicles because of their thermoresponsive behavior. pNIPAM loading/release performances are directly affected by the gel network structure. Therefore, there is a need with the approaches for accurate design of 3D pNIPAM assemblies with the structure ordered at the nanoscale. This study demonstrates size-selective spontaneous loading of macromolecules (dextrans 10-500 kDa) into pNIPAM microgels by microgel heating from 22 to 35 °C (microgels collapse and trap dextrans) followed by the dextran release upon further cooling down to 22 °C (microgels swell back) . This temperature-mediated behavior is fully reversible. The structure of pNIPAM microgels was tailored via hard templating and cross-linking of the hydrogel using sacrificial mesoporous cores of vaterite CaCO3 microcrystals. In addition, the fabrication of hollow thermoresponsive pNIPAM microshells has been demonstrated, utilizing vaterite microcrystals that had narrower pores. The proposed approach for heating-triggered encapsulation and cooling-triggered release into/from pNIPAM microgels may pave the ways for applications of pNIPAM hydrogels for skin and transdermal cooling-responsive drug delivery in the future.

CaCO3 crystals as versatile carriers for controlled delivery of antimicrobials.

CaCO3 crystals have been known for a long time as naturally derived and simply fabricated nano(micro)-sized materials able to effectively host and release various molecules. This review summarises the use of CaCO3 crystals as versatile carriers to host, protect and release antimicrobials, offering a strong tool to tackle antimicrobial resistance, a serious global health problem. The main methods for the synthesis of CaCO3 crystals with different properties, as well as the approaches for the loading and release of antimicrobials are presented. Finally, prospects to utilize the crystals in order to improve the therapeutic outcome and combat antimicrobial resistance are highlighted. Ultimately, this review intends to provide an in-depth overview of the application of CaCO3 crystals for the smart and controlled delivery of antimicrobial agents and aims at identifying the advantages and drawbacks as well as guiding future works, research directions and industrial applications.

Polyelectrolyte Multilayer Capsule (PEMC)-Based Scaffolds for Tissue Engineering.

Tissue engineering (TE) is a highly multidisciplinary field that focuses on novel regenerative treatments and seeks to tackle problems relating to tissue growth both in vitro and in vivo. These issues currently involve the replacement and regeneration of defective tissues, as well as drug testing and other related bioapplications. The key approach in TE is to employ artificial structures (scaffolds) to support tissue development; these constructs should be capable of hosting, protecting and releasing bioactives that guide cellular behaviour. A straightforward approach to integrating bioactives into the scaffolds is discussed utilising polyelectrolyte multilayer capsules (PEMCs). Herein, this review illustrates the recent progress in the use of CaCO3 vaterite-templated PEMCs for the fabrication of functional scaffolds for TE applications, including bone TE as one of the main targets of PEMCs. Approaches for PEMC integration into scaffolds is addressed, taking into account the formulation, advantages, and disadvantages of such PEMCs, together with future perspectives of such architectures.

Encapsulation of Low-Molecular-Weight Drugs into Polymer Multilayer Capsules Templated on Vaterite CaCO3 Crystals.

Polyelectrolyte multilayer capsules (PEMCs) templated onto biocompatible and easily degradable vaterite CaCO3 crystals via the layer-by-layer (LbL) polymer deposition process have served as multifunctional and tailor-made vehicles for advanced drug delivery. Since the last two decades, the PEMCs were utilized for effective encapsulation and controlled release of bioactive macromolecules (proteins, nucleic acids, etc.). However, their capacity to host low-molecular-weight (LMW) drugs (<1-2 kDa) has been demonstrated rather recently due to a limited retention ability of multilayers to small molecules. The safe and controlled delivery of LMW drugs plays a vital role for the treatment of cancers and other diseases, and, due to their tunable and inherent properties, PEMCs have shown to be good candidates for smart drug delivery. Herein, we summarize recent progress on the encapsulation of LMW drugs into PEMCs templated onto vaterite CaCO3 crystals. The drug loading and release mechanisms, advantages and limitations of the PEMCs as LMW drug carriers, as well as bio-applications of drug-laden capsules are discussed based upon the recent literature findings.

Temperature Window for Encapsulation of an Enzyme into Thermally Shrunk, CaCO3 Templated Polyelectrolyte Multilayer Capsules.

Encapsulation of enzymes allows to preserve their biological activities in various environmental conditions, such as exposure to elevated temperature or to proteases. This is particularly relevant for in vivo applications, where proteases represent a severe obstacle to maintaining the activity of enzymes. Polyelectrolyte multilayer capsules are suitable for enzyme encapsulation, where CaCO3 particles and temperature-dependent capsule formation are the best templates and the most adequate method, respectively. In this work, these two areas are combined and, ALP (alkaline phosphatase), which is a robust and therapeutically relevant enzyme, is encapsulated into thermally shrunk polyelectrolyte multilayer (PDADMAC/PSS)4 capsules templated on calcium carbonate particles (original average diameter: ≈3.5 µm). The activity of the encapsulated enzyme and the optimal temperature range for encapsulation are investigated. The enzymatic activity is almost four times higher upon encapsulation when the temperature range for encapsulation is situated just above the glass transition temperature (40 °C), while its optimal conditions are dictated, on the one hand, by the enzyme activity (better at lower temperatures) and, on the other hand, by the size and mechanical properties of capsules (better at higher temperatures).