Chitosan hybrid nanomaterials: A study on interaction with biomimetic membranes.

This study examined the influence of nanomaterials (NMs) on the organization of membrane lipids and the resulting morphological changes. The cell plasma membrane is heterogeneous, featuring specialized lipid domains in the liquid-ordered (Lo) phase surrounded by regions in the liquid-disordered (Ld) phase. We utilized model membranes composed of various lipids and lipid mixtures in different phase states to investigate the interactions between the NMs and membrane lipids. Specifically, we explored the interactions of pure chitosan (CS) and CS-modified nanocomposites (NCs) with ZnO, CuO, and SiO2 with four lipid mixtures: egg-phosphatidylcholine (EggPC), egg-sphingomyelin/cholesterol (EggSM/Chol), EggPC/Chol, and EggPC/EggSM/Chol, which represent the coexistence of Ld, Lo, and Ld/Lo, respectively. The data show that CS NMs increase the membrane lipid order at glycerol level probed by Laurdan spectroscopy. Additionally, the interaction of CS-based NMs with membranes leads to an increase in bending elasticity modulus, zeta potential, and vesicle size. The lipid order changes are most significant in the highly fluid Ld phase, followed by the Lo/Ld coexistence phase, and are less pronounced in the tightly packed Lo phase. CS NMs induced egg PC vesicle adhesion, fusion, and shrinking. In heterogeneous Lo/Ld membranes, inward invaginations and vesicle shrinking via the Ld phase were observed. These findings highlight mechanisms involved in CS NM-lipid interactions in membranes that mimic plasma membrane heterogeneity.

Anti-Biofilm Perspectives of Propolis against Staphylococcus epidermidis Infections.

Staphylococcus epidermis has emerged as the main causative agent of medical device-related infections. Their major pathogenicity factor lies in its ability to adhere to surfaces and proliferate into biofilms, which increase their resistance to antibiotics. The main objective of this study was to evaluate the use and the mechanism of action of an ethanolic extract of Spanish propolis (EESP) as a potential alternative for preventing biofilm-related infections caused by S. epidermidis. The chemical composition of propolis is reported and its antibacterial activity against several strains of S. epidermidis with different biofilm-forming capacities evaluated. The influence of sub-inhibitory concentrations (sub-MICs) of EESP on their growth, physicochemical surface properties, adherence, and biofilm formation were studied. EESP interferes with planktonic cells, homogenizing their physicochemical surface properties and introducing a significant delay in their growth. The adherence and biofilms at the EESP concentrations investigated were decreased up to 90.5% among the strains. Microscopic analysis indicated that the planktonic cells that survived the treatment were the ones that adhere and proliferate on the surfaces. The results obtained suggest that the EESP has a high potential to be used as an inhibitor of both the adhesion and biofilm formation of S. epidermidis.

Cation-induced morphological transitions and aggregation of thermoresponsive PNIPAM nanogels.

Poly(N-isopropylacrylamide) (PNIPAM) nanogels are promising responsive colloidal particles that can be used in pharmaceutical applications as drug carriers. This work investigates the temperature-dependent morphological changes and agglomeration of PNIPAM nanogels in the presence of mono- and multi-valent cationic electrolytes. We described the deswelling, flocculation, thermal reversibility behaviour and aggregated morphology of PNIPAM nanogels over a range of electrolyte concentrations and temperatures revealing the critical transition points from stable suspension to spontaneous agglomeration. We demonstrated that the flocculating ability and the rate of aggregate formation follow the order of deswelling behaviour. Transmission electron microscopy and atomic force microscopy analysis revealed the presence of a shell-like layer with varying density in the multivalent electrolyte solutions when compared to those in aqueous medium. We identified a concentration threshold of the thermally induced reversible aggregation/dispersion for the PNIPAM nanogels in the presence of Na+ and K+ ions at 10 mM, for Mg2+ and Ca2+ ions at 1 mM and for Al3+ ions at 0.1 mM concentrations. Such concentration thresholds indicated the effective destabilization of the electrolyte system with multivalency following the Schulze-Hardy rule. Our findings were supported by applying a Debye screening model that accounts for the shielding effect of multivalent cationic electrolytes on these nanogel systems. Our experiments and the models confirmed the compression of the electric double layer as the valency and ionic strength increased, except for Al3+ at higher concentrations which seemed to disrupt the electrical double layer and cause reversal of zeta potential. Our work highlights the significant impact the presence of multivalent cations can impose on the stability and morphology of nanogels, and this understanding will help in designing responsive nanogel systems based on PNIPAM nanogels.

A rapid protocol for synthesis of chitosan nanoparticles with ideal physicochemical features.

In this research, an innovative protocol is introduced to address crucial deficiencies in the formulation of chitosan nanoparticles (Cs NPs). While NPs show potential in drug delivery systems (DDSs), their application in the clinic is hindered by various drawbacks, such as toxicity, high material costs, and time-consuming and challenging preparation procedures. Within polymer-based NPs, Cs is a plentiful natural substance derived from the deacetylation of chitin, which can be sourced from the shells of shrimp or crab. Cs NPs can be formulated using the ionic gelation technique, which involves the use of a negatively charged agent, such as tripolyphosphate (TPP), as a crosslinking agent. Even though Cs is a cost-effective and biocompatible material, the formulation of Cs NPs with the correct size and surface electrical charge (zeta potential) presents a persistent challenge. In this study, various techniques were employed to analyze the prepared Cs NPs. The size and surface charge of the NPs were evaluated using dynamic light scattering (DLS). Morphological analysis was conducted using field emission-scanning electron microscopy (FE-SEM). The chemical composition and formation of Cs NPs were investigated using Fourier transform infrared (FTIR). The stability analysis was confirmed through X-ray diffraction (XRD) analysis. Lastly, the biocompatibility of the NPs was assessed through cell cytotoxicity evaluation using the MTT assay. Moreover, here, 11 formulations with different parameters such as reaction pH, Cs:TPP ratio, type of Cs/TPP, and ultrasonication procedure were prepared. Formulation 11 was chosen as the optimized formulation based on its high stability of more than three months, biocompatibility, nanosize of 75.6 ± 18.24 nm, and zeta potential of +26.7 mV. To conclude, the method described here is easy and reproducible and can be used for facile preparation of Cs NPs with desirable physicochemical characteristics and engineering ideal platforms for drug delivery purposes.

Green Synthesis of Silver Nanoparticles using Coriandrum sativum and Murraya koenigii Leaf Extract and its Thrombolytic Activity.

BACKGROUND: Plants have been used for ages in traditional medicine, and it is exciting to perceive how recent research has recognized the bioactive compounds liable for their beneficial effects. Green synthesis of metal nanoparticles is a hastily emergent research area in nanotechnology. This study describes the synthesis of silver nanoparticles (AgNPs) using Coriandrum sativum and Murraya koenigii leaf extract and its thrombolytic activity. OBJECTIVE: The aim of the study was to determine the clot lysis activity of Coriandrum sativum and Murraya koenigii synthesized silver nanoparticles. METHODS: Leaves of Coriandrum sativum and Murraya koenigii were collected. Methanolic extraction of the plant sample was done through a Soxhlet extractor. The methanolic extract obtained from both the leaves was subjected to GC-MS analysis. The synthesized NPs from leaf extracts were monitored for analysis, where the typical X-ray diffraction pattern and its diffraction peaks were identified. 3D image of the NPs was analysed by Atomic Force Microscopy. The surface charge of nanoparticles was identified by Zeta potential. The Clot lysis activity of Coriandrum sativum and Murraya koenigii synthesized silver nanoparticles were analysed by the modified Holmstorm method. RESULTS: The thrombolytic property of the methanolic extract of plants Coriandrum sativum showed clot lysis activity at 2.5 mg/mL with 45.99% activity, and Murraya koenigii extract with 66.56% activity. The nanoparticles (Nps) from Coriandrum sativum showed clot lysis activity at 2.5 mg/mL with 58.29% activity, and NPs from Murraya koenigii with 54.04% activity. Coriandrum sativum in GC-MS exhibited 3 peaks, whereas Murraya koenigii extract showed five peaks with notable bioactive compounds. CONCLUSION: These NPs were further used for biomedical applications after being fixed by an organic encapsulation agent. The present research reveals the usefulness of Coriandrum sativum and Murraya koenigii for the environmentally friendly manufacture of silver nanoparticles.

Dynamic Light Scattering of DNA-Ligand Complexes.

Dynamic light scattering (DLS) enables the characterization of sizes and electrokinetic properties of colloids, polymers, and macromolecules. DNA is a charged semiflexible polyelectrolyte that is condensed or compacted by counterions, proteins, and other condensing agents in processes such as chromosome compaction and gene therapeutic applications. DNA condensation is closely related to charge screening since packaging requires effective neutralization of its surface negative charges. In this chapter, we describe in detail the protocol for DLS DNA-ligand complexes. As an example, we describe data for the condensation of DNA by chitosan and the measurement of size, zeta potential, and electrophoretic mobility of the DNA-ligand complex by DLS.

Engineered Mesoporous Silica-Based Nanoparticles: Characterization of Surface Properties.

Mesoporous silica-based nanomaterials have emerged as multifunctional platforms with applications spanning catalysis, medicine, and nanotechnology. Since their synthesis in the early 1990s, these materials have attracted considerable interest due to their unique properties, including high surface area, tunable pore size, and customizable surface chemistry. This article explores the surface properties of a series of MSU-type mesoporous silica nanoparticles, elucidating the impact of different functionalization strategies on surface characteristics. Through an extensive characterization utilizing various techniques, such as FTIR, Z-potential, and nitrogen adsorption porosimetry, insights into the surface modifications of mesoporous silica nanoparticles are provided, contributing to a deeper understanding of their nanostructure and related interactions, and paving the way to possible unexpected actionability and potential applications.

Cr(VI) removal performance from wastewater by microflora isolated from tannery effluents in a semi-arid environment: a SEM, EDX, FTIR and zeta potential study.

Hexavalent chromium removal from the environment remains a crucial worldwide challenge. To address this issue, microbiological approaches are amongst the straightforward strategies that rely mainly on the bacteria's and fungi's survival mechanisms upon exposure to toxic metals, such as reduction, efflux system, uptake, and biosorption. In this work, scanning electron microscopy, energy-dispersive X-ray spectrophotometry, Fourier transform infrared spectroscopy, and zeta potential measurements were used to investigate the ability of chromium adsorption by Bacillus licheniformis, Bacillus megaterium, Byssochlamys sp., and Candida maltosa strains isolated from tannery wastewater. Scanning electron microscopy combined with energy dispersive X-ray spectroscopy revealed alterations in the cells treated with hexavalent chromium. When exposed to 50 mg/L Cr6+, Bacillus licheniformis and Candida maltosa cells become rough, extracellular secretions are reduced in Bacillus megaterium, and Byssochlamys sp. cells are tightly bound and exhibit the greatest Cr weight percentage. In-depth analysis of Fourier transform infrared spectra of control and Cr-treated cells unveiled Cr-microbial interactions involving proteins, lipids, amino acids, and carbohydrates. These findings were supported by zeta potential measurements highlighting significant variations in charge after treatment with Cr(VI) with an adsorption limit of 100 mg/L Cr6+ for all the strains. Byssochlamys sp. showed the best performance in Cr adsorption, making it the most promising candidate for treating Cr-laden wastewater.

Preparation and Mechanism of Shale Inhibitor TIL-NH2 for Shale Gas Horizontal Wells.

In this study, a new polyionic polymer inhibitor, TIL-NH2, was developed to address the instability of shale gas horizontal wells caused by water-based drilling fluids. The structural characteristics and inhibition effects of TIL-NH2 on mud shale were comprehensively analyzed using infrared spectroscopy, NMR spectroscopy, contact angle measurements, particle size distribution, zeta potential, X-ray diffraction, thermogravimetric analysis, and scanning electron microscopy. The results demonstrated that TIL-NH2 significantly enhances the thermal stability of shale, with a decomposition temperature exceeding 300 °C, indicating excellent high-temperature resistance. At a concentration of 0.9%, TIL-NH2 increased the median particle size of shale powder from 5.2871 µm to over 320 µm, effectively inhibiting hydration expansion and dispersion. The zeta potential measurements showed a reduction in the absolute value of illite's zeta potential from -38.2 mV to 22.1 mV at 0.6% concentration, highlighting a significant decrease in surface charge density. Infrared spectroscopy and X-ray diffraction confirmed the formation of a close adsorption layer between TIL-NH2 and the illite surface through electrostatic and hydrogen bonding, which reduced the weakly bound water content to 0.0951% and maintained layer spacing of 1.032 nm and 1.354 nm in dry and wet states, respectively. Thermogravimetric analysis indicated a marked reduction in heat loss, particularly in the strongly bound water content. Scanning electron microscopy revealed that shale powder treated with TIL-NH2 exhibited an irregular bulk shape with strong inter-particle bonding and low hydration degree. These findings suggest that TIL-NH2 effectively inhibits hydration swelling and dispersion of shale through the synergistic effects of cationic imidazole rings and primary amine groups, offering excellent temperature and salt resistance. This provides a technical foundation for the low-cost and efficient extraction of shale gas in horizontal wells.

Deposition of Human-Serum-Albumin-Functionalized Spheroidal Particles on Abiotic Surfaces: Reference Kinetic Results for Bioparticles.

Human serum albumin (HSA) corona formation on polymer microparticles of a spheroidal shape was studied using dynamic light scattering and Laser Doppler Velocimetry (LDV). Physicochemical characteristics of the albumin comprising the zeta potential and the isoelectric point were determined as a function of pH for various ionic strengths. Analogous characteristics of the polymer particles were analyzed. The adsorption of albumin on the particles was in situ monitored by LDV. The stability of the HSA-functionalized particle suspensions under various pHs and their electrokinetic properties were also determined. The deposition kinetics of the particles on mica, silica and gold sensors were investigated by optical microscopy, AFM and quartz microbalance (QCM) under diffusion and flow conditions. The obtained results were interpreted in terms of the random sequential adsorption model that allowed to estimate the range of applicability of QCM for determining the deposition kinetics of viruses and bacteria at abiotic surfaces.

Physicochemical and Adsorption Properties of Nanoporous Activated Biocarbons from Thermochemical Treatment of Horsetail Herb.

The adsorption characteristics of novel activated biocarbons prepared from horsetail herb by physical activation (using carbon dioxide) and chemical one (using phosphoric(V) acid) in the process of simultaneous proteins immobilization in multicomponent solutions were examined. The carbon materials were characterized in terms of their porous structure, acidic-basic properties, and surface morphology. The binding mechanisms of such proteins as bovine serum albumin (BSA) and lysozyme (LSZ), differing in internal stability, were determined alone and in their blends. This was done based on the comprehensive analysis of the results of adsorption/desorption, surface, electrokinetic and stability measurements. These experiments were carried out over a wide pH range of 3-11. They included the following issues: (1) determination of the protein adsorbed/desorbed amounts on/from a surface of activated biocarbons; (2) study of the kinetics of these processes; (3) examination of the macromolecules impact on the surface charge density and zeta potential of the carbon materials; and (4) determination of the suspension stability and size of aggregates formed in the examined systems. The analysis of the obtained results indicated the differences in the binding mechanism of both proteins that is of key importance for their simultaneous immobilization on activated biocarbons surface in the soil environment.

Relationships of the Surface Charge of Low-Density Lipoprotein (LDL) with the Serum LDL-Cholesterol and Atherosclerosis Levels in a Japanese Population: The DOSANCO Health Study.

AIM: This study investigated the associations of the surface charge of low-density lipoprotein (LDL) with the serum LDL-cholesterol and atherosclerosis levels in a community-based Japanese population. METHODS: The study had a cross-sectional design and included 409 community residents aged 35-79 years who did not take medications for dyslipidemia. The potential electric charge of LDL and the zeta potential, which indicate the surface charge of LDL, were measured by laser Doppler microelectrophoresis. The correlations of the zeta potential of LDL (-mV) with the serum LDL-cholesterol levels (mg/dL), cardio-ankle vascular index (CAVI), and serum high-sensitivity C-reactive protein (hsCRP) levels (log-transformed values, mg/L) were examined using Pearson's correlation coefficient (r). Linear regression models were constructed to examine these associations after adjusting for potential confounding factors. RESULTS: A total of 201 subjects with correctly stored samples were included in the primary analysis for zeta potential measurement. An inverse correlation was observed between the LDL zeta potential and the serum LDL-cholesterol levels (r=-0.20; p=0.004). This inverse association was observed after adjusting for sex, age, dietary cholesterol intake, smoking status, alcohol intake, body mass index, and the serum levels of the major classes of free fatty acids (standardized ß=-6.94; p=0.005). However, the zeta potential of LDL showed almost no association with CAVI or the serum hsCRP levels. Similar patterns were observed in the 208 subjects with compromised samples as well as all the original 409 subjects. CONCLUSION: A higher electronegative surface charge of LDL was associated with lower serum LDL-cholesterol levels in the general Japanese population.

Highly stable, fluorescent, artificial lipoprotein nanoparticles.

Here, we report a novel kind of protein nanoparticles of 11 nm in size, which have a central protein core surrounded by two layers of lipid. One layer of the lipid was covalently attached to the protein, while the other layer has been physically assembled around the protein core. Particle synthesis is highly modular, while both the size and charge of the protein nanoparticles are controlled in a predictable manner. Circular dichroism studies of the conjugate showed that the protein secondary structure is retained, while biophysical characterizations indicated the particle purity, size, and charge. The conjugate had a high thermal stability to steam sterilization conditions at 121°C (17 psi). After labeling the protein core with few different fluorescent dyes, they were strongly fluorescent with the corresponding colors independent of their size, unlike quantum dots. They are readily digested by proteases, and these water-soluble, non-toxic, highly stable, biocompatible, and biodegradable conjugates are suitable for cell imaging and drug delivery applications.

Effect of Dielectric Constant on the Zeta Potential of Spherical Electric Double Layers.

Zeta potential refers to the electrokinetic potential present in colloidal systems, exerting significant influence on the diverse properties of nano-drug delivery systems. The impact of the dielectric constant on the zeta potential and charge inversion of highly charged colloidal particles immersed in a variety of solvents spanning from polar, such as water, to nonpolar solvents and in the presence of multivalent salts was investigated through primitive Monte Carlo (MC) model simulations. Zeta potential, ξ, is decreased with the decreasing dielectric constant of the solvent and upon further increase in the salinity and the valency of the salt. At elevated levels of salt, the colloidal particles become overcharged in all solvents. As a result, their apparent charge becomes opposite in sign to the stoichiometric charge. This reversal of charge intensifies until reaching a saturation point with further increase in salinity.

Thermal Stability of Dispersions of Amino-Functionalized Silica in Glycol and in 50-50 Aqueous Glycol.

Dispersions of amino-functionalized silica in ethylene glycol (EG) and in aqueous glycol show excellent stability at room temperature. Stability at elevated temperatures would be much desired with respect to their potential application as heat-transfer fluids. Amino-functionalized silica was dispersed in EG and in 50-50 aqueous EG by mass. HCl and acetic acid were added to enhance the positive ζ potential. The dispersions were stored at 40, 60, 80, and 100 °C for up to 28 days, and ζ potential and apparent particle radius were studied as a function of elapsed time. The particles showed a positive ζ potential in excess of 40 mV (Smoluchowski), which remained unchanged for 28 days. Such a high absolute value of ζ potential is sufficient to stabilize the dispersion against flocculation and sedimentation. The apparent particle radius in acidified dispersions was about 70 nm, and it was stable for 28 days. The particles were larger in pH-neutral dispersions. The apparent particle radius was about 80 nm in fresh dispersions and it increased on long storage at 80 and 100 °C.

Lipid-Based Nanoparticles for RNA Delivery.

Various nanoparticle-based delivery systems have been developed for the encapsulation and protection of active cargoes. Lipid nanoparticles represent one of the most widely used nanoparticle-based delivery systems for in vitro and in vivo applications, especially for the delivery of ribonucleic acid (RNA). In this chapter, a simple bulk mixing method for the encapsulation of RNA is described along with characterization techniques for measuring encapsulation efficiency and nanoparticle physicochemical properties.

Synthesis and characterization of aluminosilicate and zinc silicate from sugarcane bagasse fly ash for adsorption of aflatoxin B1.

Sugarcane bagasse fly ash, a residual product resulting from the incineration of biomass to generate power and steam, is rich in SiO2. Sodium silicate is a fundamental material for synthesizing highly porous silica-based adsorbents to serve circular practices. Aflatoxin B1 (AFB1), a significant contaminant in animal feeds, necessitates the integration of adsorbents, crucial for reducing aflatoxin concentrations during the digestive process of animals. This research aimed to synthesize aluminosilicate and zinc silicate derived from sodium silicate based on sugarcane bagasse fly ash, each characterized by a varied molar ratio of aluminum (Al) to silicon (Si) and zinc (Zn) to silicon (Si), respectively. The primary focus of this study was to evaluate their respective capacities for adsorbing AFB1. It was revealed that aluminosilicate exhibited notably superior AFB1 adsorption capabilities compared to zinc silicate and silica. Furthermore, the adsorption efficacy increased with higher molar ratios of Al:Si for aluminosilicate and Zn:Si for zinc silicate. The N2 confirmed AFB1 adsorption within the pores of the adsorbent. In particular, the aluminosilicate variant with a molar ratio of 0.08 (Al:Si) showcased the most substantial AFB1 adsorption capacity, registering at 88.25% after an in vitro intestinal phase. The adsorption ability is directly correlated with the presence of surface acidic sites and negatively charged surfaces. Notably, the kinetics of the adsorption process were best elucidated through the application of the pseudo-second-order model, effectively describing the behavior of both aluminosilicate and zinc silicate in adsorbing AFB1.

Important Factors Affecting Induction of Cell Death, Oxidative Stress and DNA Damage by Nano- and Microplastic Particles In Vitro.

We have described the influence of selected factors that increase the toxicity of nanoplastics (NPs) and microplastics (MPs) with regard to cell viability, various types of cell death, reactive oxygen species (ROS) induction, and genotoxicity. These factors include plastic particle size (NPs/MPs), zeta potential, exposure time, concentration, functionalization, and the influence of environmental factors and cell type. Studies have unequivocally shown that smaller plastic particles are more cytotoxic, penetrate cells more easily, increase ROS formation, and induce oxidative damage to proteins, lipids, and DNA. The toxic effects also increase with concentration and incubation time. NPs with positive zeta potential are also more toxic than those with a negative zeta potential because the cells are negatively charged, inducing stronger interactions. The deleterious effects of NPs and MPs are increased by functionalization with anionic or carboxyl groups, due to greater interaction with cell membrane components. Cationic NPs/MPs are particularly toxic due to their greater cellular uptake and/or their effects on cells and lysosomal membranes. The effects of polystyrene (PS) vary from one cell type to another, and normal cells are more sensitive to NPs than cancerous ones. The toxicity of NPs/MPs can be enhanced by environmental factors, including UV radiation, as they cause the particles to shrink and change their shape, which is a particularly important consideration when working with environmentally-changed NPs/MPs. In summary, the cytotoxicity, oxidative properties, and genotoxicity of plastic particles depends on their concentration, duration of action, and cell type. Also, NPs/MPs with a smaller diameter and positive zeta potential, and those exposed to UV and functionalized with amino groups, demonstrate higher toxicity than larger, non-functionalized and environmentally-unchanged particles with a negative zeta potential.

Harnessing polyelectrolyte complexes for precision cancer targeting: a comprehensive review.

Polyelectrolytes represent a unique class of polymers abundant in ionizable functional groups. In a solution, ionized polyelectrolytes can intricately bond with oppositely charged counterparts, giving rise to a fascinating phenomenon known as a polyelectrolyte complex. These complexes arise from the interaction between oppositely charged entities, such as polymers, drugs, and combinations thereof. The polyelectrolyte complexes are highly appealing in cancer management, play an indispensable role in chemotherapy, crafting biodegradable, biocompatible 3D membranes, microcapsules, and nano-sized formulations. These versatile complexes are pivotal in designing controlled and targeted release drug delivery systems. The present review emphasizes on classification of polyelectrolyte complex along with their formation mechanisms. This review comprehensively explores the applications of polyelectrolyte complex, highlighting their efficacy in targeted drug delivery strategies for combating different forms of cancer. The innovative use of polyelectrolyte complex presents a potential breakthrough in cancer therapeutics, demonstrating their role in enhancing treatment precision and effectiveness.

Synchronous improvement of methane production and digestate dewaterability in sludge anaerobic digestion by nanobubble.

The subsequence anaerobic digestion (AD) of dewatered sludge (DWS) from wastewater treatment plants necessitates an emphasis on enhancing methane production and dewaterability. The effect of different nanobubble water (NBW) on AD of DWS was investigated under mesophilic conditions. Cumulative methane production was improved by 9.0-27.8% with the addition of different NBW (Air, CO2, He, and N2). NBW improved methanogenic performance by significantly enhancing the hydrolysis of sludge AD. Results from the digestate, the capillary suction time, specific resistance to filtration, and moisture content could be decreased by 14.6-18.2%, 18.8-29.6%, and 13.6-19.5%, respectively. The addition of NBW can improve the dewaterability of digestate by reducing the digestate particle size and increasing the zeta potential of digestate. The addition of NBW significantly increased methane production and improved dewaterability in AD; Air-NBW showed the best improvement.