Scalable production of glass flakes via compression in the liquid phase

A comprehensive study on controlled shape formation with high yield of three commercially relevant SiO₂-based amorphous glasses in a stirred media mill is presented. Stressing under well-controlled conditions leads to micron-sized amorphous glass flakes with high aspect ratios. This unique result is quite contrary to the fundamental observation that comminution processes generally result in irregular shaped particles. The influences of glass composition, processing time, stirrer tip speed and grinding media size on the obtained products have been investigated. The size and shape of the obtained glass flakes have been characterized by scanning electron microscopy and atomic force microscopy. The glass flakes exhibit thicknesses as low as 155 nm while the lateral dimensions are well within the micrometer range. Our study shows that particle size reduction occurs within the first hour of grinding. Afterwards plastic deformation of the fragments, which can be accompanied by densification of the glass network, leads to the formation and further thinning of the glass flakes. To demonstrate the quality and applicability of the glass flakes interference pigments were realized by a TiO₂ coating via an aqueous titration process. The presented approach offers a simple, convenient and fully scalable top-down method to produce flake-like particles from various silica glasses. The obtained flakes are suitable substrates for further modifications and applications and the process can be transferred to other materials and glasses with tailor-made chemical compositions.     

CFD modelling and simulation of an industrial scale continuous fluidized bed roaster

In the present work, computational fluid dynamics (CFD) based modelling of an industrial scale continuous fluidised bed roaster (FBR) has been carried out to study its performance at different operating conditions, so that the sulphide-sulphur content in the product is within 0.4% at the designed feed rate of 39.75 DMT/h. Eulerian-Eulerian multiphase model, considering four granular phases and one gas phase has been implemented to investigate the velocity and mass fraction profile of the particles in the FBR. The heat and species mass balance calculations have been performed external to CFD, by dividing the roaster into several sections. The conversion of ZnS to ZnO at various sections of the roaster has been estimated using reaction kinetics under isothermal condition (1203 K). The heat liberated and possible temperature rise at each section was predicted based on the heat of reaction and sensible heat of the solid and gaseous products. The CFD model was validated with the plant data for a feed rate of 36.5 DMT/h, air flow rate of 65,000 Nm³/h and O₂ content of 21%. The proposed model predicted the sulphide-sulphur content in the product to be 0.4% for the designed feed rate of 39.75 DMT/h, when the O₂ content in the inlet air was increased to 25%.     

Predicting the flowability of alumina powder during batch grinding through the establishment of a grinding kinetic model

Grinding is one of the main unit operation in industrial processes handling powders. The particle size reduction that takes place during grinding tests, usually results in a significant change in the flow behavior of the ground powder. Up to now, a model predicting the evolution of powder flowability with grinding time, according to the operating conditions is still missing. In this paper, a methodology combining a grinding kinetic model and a flowability model involving the population-dependent granular Bond number is developed. The methodology has been applied to an alumina powder, ground in a batch ball mill. The flow function coefficient of the ground samples is measured after various grinding times in a powder shear tester. The comparison between model predictions and experimental data shows that this method allows an accurate prediction of the powder flow behavior over the first sixteen minutes of grinding.     

Metal-organic framework derived hexagonal layered cobalt oxides with {1 1 2} facets and rich oxygen vacancies: High efficiency catalysts for total oxidation of propane

A cobalt-based metal–organic framework was used as a precursor to synthesize Co₃O₄ catalysts exhibiting a hexagonal layered morphology by calcination at varying temperatures. Various characterization techniques, such as XRD, SEM, Raman, H₂-TPR, O₂-TPD and N₂ adsorption–desorption, were used to study the effects of calcination temperature on the grain size, surface area, and pore volume of the catalysts. The Co₃O₄ catalyst obtained by calcination at 350 °C (Co₃O₄-350) exhibited the highest catalytic activity for the total oxidation of propane. Furthermore, the small grain size and layered structure of Co₃O₄-350 allowed it to possess a high specific surface area, a highly exposed {1 1 2} facets, and abundant oxygen defects that facilitated a favorable low-temperature reducibility and oxygen mobility, thereby improving catalytic activity. This research offers a simple strategy for synthesis of Co₃O₄ with layered structure, highly exposed {1 1 2} facets and rich oxygen defects.     

Changes in the oxygen content,morphology, and microstructure of Mo-10Nb composite powders during mechanical alloying

The prefabrication of Mo-Nb composite powders is an effective way of improving the homogeneity of Mo-10Nb targets, which have broad application prospects in the photoelectric sensor industry. However, this aspect has been rarely addressed so far. Therefore, we prepared Mo-10Nb composite powders by mechanical alloying (MA), and investigated the effects of the experimental parameters such as the milling speed and duration on the particle morphology, size distribution, compositional homogeneity, crystallite size, inner strain, and oxygen content. High-quality Mo-10Nb composite powders with 3-μm spherical particles of narrow size distribution, homogeneous elemental distribution, and nanometric crystalline structure were obtained by implementing optimum MA parameters, viz., a milling speed of 250 rpm and duration of 36 h using an MITR QM-QX-4L omnidirectional ball mill. The mechanically alloyed Mo-10Nb composite powders were prone to oxidation when exposed to air, which led to a sharp increase in the oxygen content to ~5400 ppm. X-ray photoelectron spectroscopic analysis revealed the presence of Nb₂O₅, MoO₂, and MoO₃ on the surface of the Mo-10Nb particle. We believe that this study demonstrates an interesting strategy for the fabrication of high-quality Mo-10Nb targets.     

3D HCN nanotexture with synergistic effect of nickel and hole scavengers for enhancing photocatalytic H2 production: Role of morphology and influential parameters

Well-designed three-dimensional (3D) nanotextures of graphitic carbon nitride (g-C₃N₄), synthesized using template free single step method and mediated with nickel as a noble free metal, for solar hydrogen production, has been investigated. The photoactivity was investigated in a slurry type continuous flow photoreactor system by using different influential parameters such as hole scavengers, diffusion effects, time, and mass transfer. Compared to bulk g-C₃N₄, H₂ yield was increased with 3D hierarchical carbon nitride (HCN) nanotexture. The H₂ evolution rate was reached to 1310 µmol g^?1 h^?1 with optimized 2 % Ni loading to 3D HCN. This H₂ evolution rate was 19.8 and 24.9 times higher than it was generated using 3D HCN and g-C₃N₄, respectively. The special interlayer opening, more light penetration and suppressed charge carrier recombination were the main contributors for this photoactivity enhancement. Among the different influential parameters, lower viscosity, higher number of protons and less diffusion effects were promising to give significantly higher H₂ production. The stability of nanotextures was entirely dependent on the attached reactants over the nickel reactive sites, which was more promising for Triethanolamine (TEOA) than using methanol. This newly developed low-cost 3D HCN can be promising in solar energy conversion and other energy applications.     

Breaking behaviour and interactions in maize and soybean meal while grinding of a hammer mill

The objective of the present study was to investigate whether mixing ratio of maize and soybean meal (SBM) affects the breaking behaviour during hammer-milling in terms of the nutrient properties and in vitro digestibility of fractionated particles. Mixtures of maize and SBM with different proportions (% Maize:SBM; 0:100, 25:75, 50:50, 75:25, 100:0) were hammer milled using a 2-mm screen. The obtained powder was sieved into seven fractions with size ranges from 0.149 to 1.190 mm. Results show that energy consumption of grinding mixtures increased from 3.8 to 48.4 kJ/kg with the maize proportion increasing from zero to 100%. Mixing proportion of maize and SBM showed significant effects on nutrient content of fractionated material. For hammer milled material <595 µm, the in vitro digestibility of crude protein (CP) and organic matter (OM) of fractionated material decreased with increasing particle size. Additionally grinding fractionated particles ≥595 µm over a 1-mm sized screen before in vitro digestion analysis increased the digestibility of OM and CP. Equivalent particle size (EPS) and geometric standard deviation (GSD) of hammer milled maize and SBM and their mixtures correlated better than geometric mean diameter (GMD) to OM and CP in vitro digestibility in a linear regression model. In summary, the mixing ratio of maize and SBM had a significant effect on the breaking behaviour of ingredients and in vitro digestibility of CP and OM of the isolated fractions. Mixing ingredients before grinding is suggested in terms of saving energy consumption. The GSD/EPS of ground material should be considered while studying the effects of particle size distribution on the in vitro digestibility of nutrients.     

Recent progress in bismuth-based high Curie temperature piezo-/ferroelectric perovskites for electromechanical transduction applications

Piezo-/ferroelectric materials with high Curie temperature (T_C) are widely needed in sensors, actuators and transducers which can be used for high-temperature (HT) electromechanical transduction applications. In recent years, remarkable progress has been made in bismuth-based piezo-/ferroelectric perovskite materials (BPPs). In this article, recent progress in high T_C BPPs is reviewed. This review starts with an introduction to HT piezoelectrics and their applications. A detailed survey is then carried out on bismuth-based perovskites (BPs) with high T_C. Material synthesis, doping effects and chemical modifications of the related solid solutions are examined. Based on this analysis, the structure–property relationship of these materials is established. In addition, recent developments of BPPs for HT electromechanical transduction applications are presented and evaluated. Lastly, some main existing issues are analyzed and their possible solutions are proposed. This article provides a comprehensive overview of the research and development of BPPs and offers some prospects towards making these materials a viable resource for the design and fabrication of electromechanical transducers with unique specifications, especially, high temperature, high frequency and high power, for a wide range of technological applications.     

3D printing vertically: Direct ink writing free-standing pillar arrays

Over the last three decades, a variety of additive manufacturing techniques have gradually gained maturity and will potentially play an important role in future manufacturing industries. Among them, direct ink writing has attracted significant attention from both material and tissue engineering areas, where the colloidal ink is extruded and dispensed according to a pre-designed path, usually in the X-Y plane with suitable increments in the Z direction. Undoubtedly, this way of disassembling geometries, simple or complex, can facilitate most of the printing process. However, for one extreme case, i.e. pillar arrays, the size resolution can deviate from both nozzle and design if the common way of slicing and additive manufacturing is used. Therefore, a different printing path is required – directly depositing pillars in a converse gravitational direction. This paper gives multiple examples of printing viscoelastic colloidal ceramic and metal inks uniaxially and periodically into free-standing and height-adjustable pillar arrays. It is expected to inspire the additive manufacturing community that more versatile degrees of freedom and complex printing paths, not confined within only complex shapes, can be achieved by ink-based 3D printing.     

Designing amorphous formulations of polyphenols with naringin by spray-drying for enhanced solubility and permeability

Naringin (NAR), a major flavanone (FVA) glycoside, is a component of food mainly obtained from grapefruit. We used NAR as a food additive to improve the solubility and permeability of hydrophobic polyphenols used as supplements in the food industry. The spray-dried particles (SDPs) of NAR alone show an amorphous state with a glass transition temperature (T_g) at 93.2 °C. SDPs of hydrophobic polyphenols, such as flavone (FVO), quercetin (QCT), naringenin (NRG), and resveratrol (RVT) were prepared by adding varying amounts of NAR. All SDPs of hydrophobic polyphenols with added NAR were in an amorphous state with a single T_g, but SDPs of hydrophobic polyphenols without added NAR showed diffraction peaks derived from each crystal. The SDPs with NAR could keep an amorphous state after storage at a high humidity condition for one month, except for SDPs of RVT/NAR. SDPs with NAR enhanced the solubility of hydrophobic polyphenols, especially NRG solubility, which was enhanced more than 9 times compared to NRG crystal. The enhanced solubility resulted in the increased membrane permeability of NRG. The antioxidant effect of the hydrophobic NRG was also enhanced by the synergetic effect of NAR. The findings demonstrated that NAR could be used as a food additive to enhance the solubility and membrane permeability of hydrophobic polyphenols.     

Steel slag waste combined with melamine pyrophosphate as a flame retardant for rigid polyurethane foams

To explore the potential application of industrial waste, steel slag powder in combination with melamine pyrophosphate (MPP) was adopted to improve the flame retardancy of rigid polyurethane foam (RPUF). The incorporation of steel slag slightly reduced the thermal conductivity of the resulting flame-retardant RPUF samples. The addition of MPP and/or steel slag did not significantly alter the thermal stability in terms of T_-10% and T_max but did obviously increase the T_-50% value, suggesting the improved thermal resistance of the residues. The coaddition of MPP and steel slag into RPUF resulted in higher LOI values and lower peak heat release rates than the samples incorporating either MPP or steel slag alone. The superior flame retardancy could be attributed to MPP promoting char formation, which then acted as a barrier at the beginning of RPUF thermal decomposition; simultaneously, the thermally stable inorganics in the steel slag powder strengthened the thermal resistance of this char layer.     

Investigation of the molecular state of 4-aminosalicylic acid in matrix formulations for dry powder inhalers using solid-state fluorescence spectroscopy of 4-dimethylaminobenzonitrile

Carrier-free method is an alternative approach for dry powder inhaler (DPI) formulations, which overcome poor drug mobility and distribution. Here we investigated the properties of an active pharmaceutical ingredient (API) within composite particles. We used highly-branched cyclic dextrin (HBCD) as the excipient matrix that was prepared using a spray-drying technique. 4-Aminosalicylic acid (4-ASA) and 4-dimethylaminobenzonitrile (DMABN) were selected as a hydrophilic second-line antitubercular agent and a surrogate for 4-ASA as a model compound, respectively. The spray-dried particles (SDPs) containing 4-ASA or DMABN with HBCD had geometric median diameters (D₅₀) of 2.34 ± 0.07 μm and 2.26 ± 0.10 μm, respectively. Further, the in vitro aerodynamic properties were similar for SDPs containing 4-ASA and DMABN with HBCD. To determine the properties of APIs within composite particles, we performed solid-state fluorescence spectroscopy of DMABN. As a candidate excipient, hydroxypropyl methylcellulose (HPMC) was compared to HBCD. We determined the intensity ratio of twisted intramolecular charge transfer (TICT) emission to locally excited emission within the excipient matrix environment. The HBCD matrix environment was better than HPMC to trigger a more robust TICT reaction of DMABN. A potent state-changing interaction of APIs occurred in the HBCD matrix environment versus another excipient environment.     

Particle tracking microrheology of cancer cells in living subjects

Cumulative evidence shows that microenvironmental conditions play a significant role in the regulation of cell functions, and how cells respond to these conditions are of central importance to regenerative medicine and cancer cell response to therapeutics. Here, we develop a new method to examine cell mechanical properties by analyzing the motion of nanoparticles in living in mice, combining particle tracking with intravital microscopy. This method directly examines the mechanical response of breast carcinoma cells and normal breast epithelial cells under intravital microenvironments. Our results show both carcinoma and normal cells display significantly reduced compliance (less deformability) in vivo compared to the same cells cultured in 2D, in both sparse and confluent conditions. While the compliance of the normal cells remains steady over time, the compliance of carcinoma cells decreases further as they form tumor-like architectures. Integrating the cancer cells into spheroids embedded in 3D collagen matrices in part redirected the mechanical response to a state closer to the in vivo setting. Overall, our study demonstrates that the microenvironment is a crucial regulator of cell mechanics and the intravital particle tracking method can provide novel insights into the role of cell mechanics in vivo.     

Synthesis,structure, and mechanical response of Cr0.26Fe0.24Al0.5 and Cr0.15Fe0.14Al0.30Cu0.13Si0.28 nanocrystallite entropy alloys

The present research work has concentrated to synthesize nanocrystalline (NC) Cr_0.26Fe_0.24Al_0.5 (medium entropy alloy, 3E-MEA) and Cr_0.15Fe_0.14Al_0.30Cu_0.13Si_0.28 (high-entropy alloy, 5E-HEA) non-equiatomic (equal weight fraction) alloys through mechanical alloying (MA); which studied the influence of entropy effect on structural properties, microstructural characterization, and mechanical behaviour. Further, the same non-equiatomic ratio of two coarse grain alloys (CGAs) was manufactured by conventional powder metallurgy (PM) route (blending method, 3E-CGA, 5E-CGA) for comparison. All synthesized powders were hot-pressed (HPed) at 723 k for 30 min subsequently mechanical properties in terms of compressive stress-strain and hardness were examined. The samples of as-milled powders, HPed, and fractured were investigated using X-ray diffraction (XRD) and advanced electron microscopes. The HPed sample of 3E-MEA of Cr_0.26Fe_0.24Al_0.5 produced 94% BCC and 6% FCC crystal structures due to more dissolution of Al atoms in the stronger bonding atoms of Cr-Fe lattice. Whereas 5E-HEA of Cr_0.15Fe_0.14Al_0.30Cu_0.13Si_0.28 sample has exhibited 72.1% FCC phase and 27.9% BCC phase due to balance between the dissolution of FCC elements (Al, Cu, Si) and BCC elements (Cr, Fe). Further, 3E-MEA and 5E-HEA have exhibited the ultimate compressive strength (UCS) of 1278 ± 6.75 MPa and 2060 ± 2.8 MPa respectively whereas the corresponding conventionally blended alloys produced 268 ± 5 MPa and 615 ± 3 MPa for 3E-CGA and 6E-CGA respectively. Vicker’s hardness strength (VHS) of 5E-HEA of Cr_0.15Fe_0.14Al_0.30Cu_0.13Si_0.28 has exhibited 68% more when compared to 3E-MEA of Cr_0.26Fe_0.24Al_0.5, 3.26 times higher compared to blended alloys. Further, several strengthening mechanisms on the mechanical behaviour of MEA and HEA were investigated in which dislocation strengthening mechanisms followed by solid solution strengthening mechanisms have influenced more as compared to grain boundary strengthening mechanisms.     

Evaluation of particle recovery from microalgae

Coccoliths are micro-structured biomineral particles found in cell protective covering layers of coccolithophore species. They are mainly composed of CaCO₃ and their individual crystal entities are arranged in such a way that they construct complex and unique structures. This complexity is found down to the individual particle level and appears to have promising properties to offer. This study focuses on the essential step prior to any kind of implementation, which is the recovery of the material. It summarizes cleaning protocols found in literature, compares them for the first time for the same freshly cultivated material and addresses challenges that still need to be overcome. Further, it highlight the advantages and disadvantages of the best cleaning protocols, suggests optimizations with promising results and uses size distribution measurements to analyse the recovery efficiency. To that end, further characterization techniques, new for coccoliths, are introduced and used to improve our current knowledge of the particles behaviour.     

pH-responsive dual-functional hydrogel integrating localized delivery and anti-cancer activities for highly effective therapy in PDX of OSCC

As one of the most promising localized drug delivery systems for enhancing therapeutic efficacy and reducing systemic toxicity, supramolecular hydrogels self-assembled from natural products have recently attracted tremendous attention. However, the intricate drug loading process, limited drug entrapment efficacy, and lack of stimulus responsiveness considerably impede their potential for biological applications and raise the need for advanced hydrogel-based delivery systems. Therefore, the development of updated materials that integrate localized delivery and drug activity into a single system is extremely desired and has great potential to overcome the aforementioned shortcomings. In this study, a pH-responsive dual-functional isoG-based supramolecular hydrogel with both localized delivery and anti-cancer activity in one molecule is successfully developed in one pot by following a simple and green procedure. The isoguanosine-phenylboronic-guanosine (isoGPBG) hydrogel exhibits exceptional stability (more than one year), outstanding pH-responsiveness and excellent sustained release capability. Both in vitro and in vivo experiments demonstrate that the isoGPBG hydrogel not only shows acceptable biocompatibility and biodegradability but also significantly inhibit tumor growth (approximately 60% inhibition of tumor growth) and improve overall survival, especially in preclinical patient-derived xenograft (PDX) model of oral squamous cell carcinoma (OSCC). Therefore, the isoGPBG hydrogel, to the best of our knowledge, is the first example of pH-responsive dual-functional isoG-based supramolecular hydrogel integrating localized delivery and anti-cancer activity in one molecule. It is implied that the isoGPBG hydrogel could act as a smart dual-functional localized delivery system in the future for clinical cancer therapy.     

Recent breakthroughs and perspectives of high-energy layered oxide cathode materials for lithium ion batteries

Ni-rich layered oxides (NRLOs) and Li-rich layered oxides (LRLOs) have been considered as promising next-generation cathode materials for lithium ion batteries (LIBs) due to their high energy density, low cost, and environmental friendliness. However, these two layered oxides suffer from similar problems like capacity fading and different obstacles such as thermal runaway for NRLOs and voltage decay for LRLOs. Understanding the similarities and differences of their challenges and strategies at multiple scales plays a paramount role in the cathode development of advanced LIBs. Herein, we provide a comprehensive review of state-of-the-art progress made in NRLOs and LRLOs based on multi-scale insights into electrons/ions, crystals, particles, electrodes and cells. For NRLOs, issues like structure disorder, cracks, interfacial degradation and thermal runaway are elaborately discussed. Superexchange interaction and magnetic frustration are blamed for structure disorder while strains induced by universal structural collapse result in issues like cracks. For LRLOs, we present an overview of the origin of high capacity followed by local crystal structure, and the root of voltage hysteresis/decay, which are ascribed to reduced valence of transition metal ions, phase transformation, strains, and microstructure degradation. We then discuss failure mechanism in full cells with NRLO cathode and commercial challenges of LRLOs. Moreover, strategies to improve the performance of NRLOs and LRLOs from different scales such as ion-doping, microstructure designs, particle modifications, and electrode/electrolyte interface engineering are summarized. Dopants like Na, Mg and Zr, delicate gradient concentration design, coatings like spinel LiNi_0.5Mn_1.5O₄ or Li₃PO₄ and novel electrolyte formulas are highly desired. Developing single crystals for NRLOs and new crystallographic structure or heterostructure for LRLOs are also emphasized. Finally, remaining challenges and perspectives are outlined for the development of NRLOs and LRLOs. This review offers fundamental understanding and future perspectives towards high-performance cathodes for next-generation LIBs.     

Synergistic antidiabetic activity of ZnO nanoparticles encompassed by Urtica dioica extract

The present study aimed at producing ZnO nanoparticles using the leaf extract of nettle (Urtica dioica) as a medicinally valuable plant to maximize the antidiabetic property of ZnO while excluding the chemical pollution from the synthesis process. The properties of the ZnO-extract sample were uncovered by various techniques and compared to that produced without the extract (ZnO). The results of the surface, optical, and thermal studies disclosed the presence of the extract biomolecules over the ZnO-extract sample and was further confirmed by GC–MS analysis. The ZnO-extract was intraperitoneally injected to alloxan-induced diabetic rats and the effects on the serum levels of fasting blood glucose, insulin, high-density lipoprotein cholesterol, total cholesterol, and total triglyceride were assessed. The obtained results were then compared with the effects of ZnO, nettle leaf extract, and insulin on the same factors. Among all the examined treatments, the best antidiabetic performance was obtained in the rats treated by ZnO-U. dioica extract mainly owing to the great synergistic interaction between its constituents.     

Improving hydrothermal performance of hybrid nanofluid in double tube heat exchanger using tapered wire coil turbulator

Tapered wire coil insert is proposed as a novel enhancer in the double tube heat exchanger and experimental studies on Al₂O₃ + MgO hybrid nanofluid flowing under the turbulent condition are performed to investigate the hydrothermal characteristics. Effects of using tapered wire coil turbulator and hybrid nanofluid on the hydrothermal behaviors are examined for different coil configurations (Converging (C) type, Diverging (D) type and Conversing-Diverging (C-D) type) and hybrid nanofluid inlet temperatures and volume flow rates. Results show that D-type wire coil insert promotes better hydrothermal performance as compared to C-type and C-D type. Nusselt number and friction factor of hybrid nanofluid using D-type, C-D type and C-type wire coil inserts enhance up to 84%, 71% and 47%, and 68%, 57% and 46%, respectively than that of water in tube without insert. The entropy generation of hybrid nanofluid is lower than that of base fluid in all cases. The thermal performance factor for hybrid nanofluid is found more than one with all inserts. The thermal performance factor is observed a maximum of 1.69 for D-type coil. The study reveals that the hybrid nanofluid and tapered wire coil combination is promising option for improving the hydrothermal characteristics of double pipe heat exchanger.     

Three-dimensional periodic structures of gold nanoclusters in the interstices of sub-100 nm polymer particles toward surface-enhanced Raman scattering

Regularly ordered polymer nanoparticle (PNP) assemblies incorporating gold nanoparticle (Au NP) clusters into the PNP interstices were fabricated by a simultaneous deposition of PNPs and Au NPs on a glass substrate. Monodisperse PNPs with an average size of 66 nm were employed as a template in the co-assembly to create the sub-100 nm periodic Au nanostructures on the substrate. First, mono-layering of PNP array with incorporation of 14 nm Au NPs was performed by a drop-casting to examine the number ratio of Au NPs to PNPs for multi-layering. Absorption spectra of the mono-layered co-assemblies of PNPs and Au NPs were employed to characterize the clustered state of Au NPs in the interstices of mono-layered PNPs. The number ratio suitable for homogeneous incorporation of Au NPs clustered in the interstice was found to be ranged from 6 to 8 in the characterization. Then, multi-layered co-assemblies of PNPs and clustered Au NPs were fabricated by a vertical deposition method with the Au NP number ratio of 8 to PNPs. Lifting rate of the substrate on which the PNPs were deposited was varied in the vertical deposition method to tune the film thickness of NP co-assembly. A decrease in the lifting rate to 1 μm/s could thicken the film to 0.71 μm corresponding to 13 layers of PNPs, resulting in the fabrication of periodic structures of Au NP clusters with a high packing density. Signal-to-noise ratio in the Raman measurement using p-mercaptobenzoic acid as a target molecule was successfully enhanced by multi-layering of the co-assembly, indicating that Au NP clusters were homogeneously incorporated into the interstices of PNPs in the co-assemblies.