Biomedical Applications Research Articles

Enzymatic Production of Trehalose and Trehalulose by Immobilized Thermostable Trehalose Synthase.

Trehalose, a versatile disaccharide renowned for its unique physical and chemical properties, finds extensive application in the food, pharmaceutical, and cosmetic industries. While conventional extraction methods face challenges, enzymatic conversion offers a promising avenue for the industrial production of trehalose. This study delves into a novel synthetic approach utilizing a recombinant enzyme, merging the thermostable trehalose synthase domain from Thermus thermophiles with a cellulose binding domain. Immobilization of this enzyme on cellulose matrices enhances stability and facilitates product purification, opening avenues for efficient enzymatic synthesis. Notably, the engineered enzyme demonstrates additional activity, converting sucrose into trehalulose. This dual functionality, combined with immobilization strategies, holds immense potential for scalable and cost-effective production of trehalose and trehalulose, offering promising prospects in various industrial and biomedical applications.

Sophorolipid: An Effective Biomolecule for Targeting Microbial Biofilms.

Biofilms are microbial aggregates encased in a matrix that is attached to biological or nonbiological surfaces and constitute serious problems in food, medical, and marine industries and can have major negative effects on both health and the economy. Biofilm's complex microbial community provides a resistant environment that is difficult to eradicate and is extremely resilient to antibiotics and sanitizers. There are various conventional techniques for combating biofilms, including, chemical removal, physical or mechanical removal, use of antibiotics and disinfectants to destroy biofilm producing organisms. In contrast to free living planktonic cells, biofilms are very resistant to these methods. Hence, new strategies that differ from traditional approaches are urgently required. Microbial world offers a wide range of effective "green" compounds such as biosurfactants. They outperform synthetic surfactants in terms of biodegradability, superior stabilization, and reduced toxicity concerns. They also have better antiadhesive and anti-biofilm capabilities which can be used to treat biofilm-related problems. Sophorolipids (SLs) are a major type of biosurfactants that have gained immense interest in the healthcare industries because of their antiadhesive and anti-biofilm properties. Sophorolipids may therefore prove to be attractive substances that can be used in biomedical applications as adjuvant to other antibiotics against some infections through growth inhibition and/or biofilm disruption.

Reactive oxygen species-mediated organic long-persistent luminophores light up biomedicine: from two-component separated nano-systems to integrated uni-luminophores.

Organic luminophores have been widely utilized in cells and in vivo fluorescence imaging but face extreme challenges, including a low signal-to-noise ratio (SNR) and even false signals, due to non-negligible background signals derived from real-time excitation lasers. To overcome these challenges, in the last decade, functionalized organic long-persistent luminophores have gained much attention. Such luminophores could not only overcome the biological toxicity of inorganic long-persistent luminescent materials (metabolic toxicity and leakage risk of inorganic heavy metals), but also continue to emit long-persistent luminescence after removing the excitation source, thus effectively improving imaging quality. More importantly, organic long-persistent luminophores have good structure tailorability for the construction of activable probes, which is favorable for biosensing. Recently, the development of reactive oxygen species (ROS)-mediated long-persistent (ROSLP) luminophores (especially organic small-molecule ROSLP luminophores) is still in the rising stage. Notably, ROSLP luminophores for in vivo imaging have experienced from two-component separated nano-systems to integrated uni-luminophores, which obtained gradually better designability and biocompatibility. In this review, we summarize the progress and challenges of organic long-persistent luminophores, focusing on their development history, long-persistent luminescence working mechanisms, and biomedical applications. We hope that these insights will help scientists further develop functionalized organic long-persistent luminophores for the biomedical field.

Non-contact photoacoustic imaging with a silicon photonics-based Laser Doppler Vibrometer

Photoacoustic imaging has emerged as a powerful, non-invasive modality for various biomedical applications. Conventional photoacoustic systems require contact-based ultrasound detection and expensive, bulky high-power lasers for the excitation. The use of contact-based detectors involves the risk of contamination, which is undesirable for most biomedical applications. While other non-contact detection methods can be bulky, in this paper, we demonstrate a proof-of-concept experiment for compact and contactless detection of photoacoustic signals on silicone samples embedded with ink-filled channels. A silicon photonics-based Laser Doppler Vibrometer (LDV) detects the acoustic waves excited by a compact pulsed laser diode. By scanning the LDV beam over the surface of the sample, 2D photoacoustic images were reconstructed of the sample.

Limitations of effective medium models for tissue phantoms in the THz frequency range

The absorption of terahertz (THz) radiation by water molecules facilitates its application to several biomedical applications such as cancer detection. Therefore, it is critical for the THz technologies to be characterised with water content in a sample. In this paper, we analyse gelatine phantoms in the THz frequency range, with continuously varying hydration levels as they dry over time. Water molecules in close proximity to the protein molecule, termed ‘bound water’, feature properties different from the ‘free water’ molecules at larger distances. We find that a common model for predicting electromagnetic properties of phantoms and tissue samples, which assumes that only the free water varies with hydration while the bound water remains constant, does not agree well with measured results. To gain insight into this behaviour, we simultaneously measured the phantom in Raman spectroscopy, which shows a continuously varying concentration of bound water with hydration level. It follows from this investigation, that the permittivity contributions of neither the biomolecules nor water are expected to be linear with water density. This means that the often used, simple effective medium model will not be accurate for many biological tissues or phantoms.

Mechanistic understanding of enhancing bioactivity via bio-ionic liquid functionalization of biomaterials

The selection of biomaterials is a pivotal criterion for designing tissue engineering scaffolds, crucial for diverse biomedical applications. While natural biomaterials offer inherent bioactivity and biocompatibility, fine-tuning their properties for specific applications poses challenges. Conversely, synthetic biomaterials, while highly customizable, are often bio-inert. Functionalizing materials with bioactive molecules can improve cell-material interactions. Prevailing approaches utilizing RGD peptides or natural/synthetic composites have drawbacks including cost and immunogenicity. Bio-ionic liquids (BILs), a class of biocompatible ionic liquids derived from natural or synthetic biomolecules, offer a potential alternative. Herein, we present the biofunctionalization of a bio-inert polyethylene glycol diacrylate (PEGDA) with an acrylated choline-based BIL (“BioPEG”) as a method for enhancing the bioactivity of biomaterials. Cell studies using mouse myoblast C2C12 and human mesenchymal stem cells (hMSCs) revealed the excellence of BioPEG hydrogels in promoting cell attachment, growth, proliferation, and differentiation. Computational molecular dynamics (MD) simulations elucidated the mechanism by which BIL facilitates cell interaction with PEGDA. 3D printability of BioPEG hydrogel was showcased using a digital light processing (DLP) bioprinter, with printed scaffolds demonstrating excellent cytocompatibility. Collectively, these findings present BIL as a versatile bioactive agent for augmenting cell adhesion to materials, thus enriching the repertoire of biomaterials for advanced tissue engineering.

Bubble-Inspired Multifunctional Magnetic Microrobots for Integrated Multidimensional Targeted Biosensing.

Microrobots possessing multifunctional integration are desired for therapeutics and biomedicine applications. However, existing microrobots with desired functionalities need to be fabricated through complex procedures due to their constrained volume, limited manufacturing processes, and lack of effective in vivo observation methods. Inspired by bubbles exhibiting various abilities, we report magnetic air bubble microrobots with simpler structures to simultaneously integrate multiple functions, including microcargo delivery, multimode locomotion, imaging, and biosensing. Contributed by buoyancy and magnetic actuation to overcome obstacles, flexible three-dimensional locomotion is implemented, guaranteeing the integrity of micro-objects adsorbed on the surface of the air bubble microrobot. Introducing air microbubbles enhances the ultrasound imaging capability of microrobots in the vascular system of mice in vivo, facilitating ample medical applications. Moreover, air-liquid reactions endow microrobots with rapid pH biosensing. This work provides a unique strategy to utilize relatively simple air bubbles to achieve the complex functions of microrobots for biomedical applications.

Automated model discovery for textile structures: The unique mechanical signature of warp knitted fabrics

Textile fabrics have unique mechanical properties, which make them ideal candidates for many engineering and medical applications: They are initially flexible, nonlinearly stiffening, and ultra-anisotropic. Various studies have characterized the response of textile structures to mechanical loading; yet, our understanding of their exceptional properties and functions remains incomplete. Here we integrate biaxial testing and constitutive neural networks to automatically discover the best model and parameters to characterize warp knitted polypropylene fabrics. We use experiments from different mounting orientations, and discover interpretable anisotropic models that perform well during both training and testing. Our study shows that constitutive models for warp knitted fabrics are highly sensitive to an accurate representation of the textile microstructure, and that models with three microstructural directions outperform classical orthotropic models with only two in-plane directions. Strikingly, out of 214=16,384 possible combinations of terms, we consistently discover models with two exponential linear fourth invariant terms that inherently capture the initial flexibility of the virgin mesh and the pronounced nonlinear stiffening as the loops of the mesh tighten. We anticipate that the tools we have developed and prototyped here will generalize naturally to other textile fabrics–woven or knitted, weft knit or warp knit, polymeric or metallic–and, ultimately, will enable the robust discovery of anisotropic constitutive models for a wide variety of textile structures. Beyond discovering constitutive models, we envision to exploit automated model discovery for the generative material design of wearable devices, stretchable electronics, and smart fabrics, as programmable textile metamaterials with tunable properties and functions. Our source code, data, and examples are available at https://github.com/LivingMatterLab/CANN. Statement of significanceTextile structures are rapidly gaining popularity in many biomedical applications including tissue engineering, wound healing, and surgical repair. A precise understanding of their unique mechanical properties is critical to tailor them to their specific functions. Here we integrate mechanical testing and machine learning to automatically discover the best models for knitted polypropylene fabrics. We show that warp knitted fabrics possess a complex symmetry with three distinct microstructural directions. Along these, the behavior is dominated by an exponential linear term that characterize the initial flexibility of the virgin mesh and the nonlinear stiffening as the loops of the fabric tighten. We expect that our technology will generalize naturally to other fabrics and enable the robust discovery of complex anisotropic models for a wide variety of textile structures.

Green Synthesis of Dracocephalum kotschyi-Coated Silver Nanoparticles: Antimicrobial, Antioxidant, and Anticancer Potentials.

BACKGROUND The environmentally friendly production of silver nanoparticles (AgNPs) has gained significant attention as a sustainable alternative to traditional chemical methods. This study focused on synthesizing AgNPs using extract of Dracocephalum kotschyi (D. kotschyi), a medicinal plant. MATERIAL AND METHODS The biosynthesis of AgNPs was monitored using UV-visible spectrophotometry. The role of phytoconstituents from D. kotschyi in stabilizing AgNPs was analyzed using Fourier-transform infrared (FTIR) spectroscopy. Dynamic light scattering (DLS) spectroscopy was used to determine the size, charge, and polydispersity of the nanoparticles, while scanning electron microscopy (SEM) was employed to assess their morphology. We evaluated the antimicrobial efficacy of the synthesized AgNPs against various bacteria, their antioxidant properties via a 2,2-Diphenyl-1-picrylhydrazyl (DPPH) assay, and their cytotoxic activity against the HeLa cervical cancer cell line. RESULTS The formation of AgNPs was indicated by a color change and the emergence of a surface plasmon resonance peak at 418 nm. The nanoparticles demonstrated significant antimicrobial, antioxidant, cytotoxic, and anticancer activities. Morphology, size, and shape analysis revealed nearly spherical particles with an average size of 43 nm. FTIR confirmed the presence of phenolic compounds in the extract, serving as reducing and capping agents. X-ray diffraction (XRD) analysis confirmed the crystalline structure of the nanoparticles. Antimicrobial assessments showed effectiveness against Escherichia coli and Staphylococcus aureus. The DPPH scavenging assay demonstrated efficient antioxidant activity, and potent apoptotic anticancer effects were observed on cervical cancer cells. CONCLUSIONS The extract of D. kotschyi was effective as a reducing agent in the environmentally friendly synthesis of AgNPs, which exhibited noteworthy antimicrobial, antioxidant, and anticancer properties. These findings suggest potential biomedical applications for the synthesized AgNPs.

Poly (lactic acid) electrospun nanofiber membranes: Advanced characterization for biomedical applications with drug loading performance studies

Traditional dressings have shortcomings such as poor moisture absorption and easy to adhere, making the development of new dressings crucial. In this work, a PLA/PVP crosslinked drug-loaded nanofiber membrane was prepared through electrospinning and ultraviolet crosslinking, with poly (lactic acid) (PLA), polyvinylpyrrolidone (PVP), and salicylic acid (SA) as starting materials. The results demonstrated that the inclusion of PVP notably boosted the viscosity and conductivity of the blend spinning solution. The roughness of the fabricated fiber was elevated, and the diameter of the fibers was more uniform. Additionally, the incorporation of PVP not only enhanced the porosity of the fiber membrane but also effectively decreased its contact angle. Notably, when the PVP content reached 40 %, the contact angle underwent a substantial reduction, decreasing significantly from 125.4° to 82.2°. The SA drug-loaded fiber membrane exhibited a notable bacteriostatic effect against Escherichia coli and Staphylococcus aureus, with its release behavior adhering to Fick's diffusion law. In the cell viability experiment, the cell proliferation rate increased from 94 % to 129 % after 3 days. This shows that the prepared membrane has good antibacterial effect and cell compatibility, which provides a theoretical basis for the construction of a new medical dressing.

Adenosine Triphosphate Inhibits Cold-Responsive Aggregation.

Adenosine triphosphate (ATP), ubiquitous in all living organisms, is conventionally recognized as a fundamental energy currency essential for a myriad of cellular processes. While its traditional role in energy metabolism requires only micromolar concentrations, the cellular content of ATP has been found to be significantly higher at the millimolar level. Recent studies have attempted to correlate this higher concentration of ATP with its nonenergetic role in maintaining protein homeostasis, leaving the investigation of ATP's nontrivial activities in biology an open question. Here, by coupling computer simulations and experiments, we uncover new insights into ATP's role as a cryoprotectant against cold-salt stress, highlighting the necessity for higher cellular ATP concentrations. We present direct evidence at charged silica interfaces, demonstrating ATP's ability to restore native intersurface interactions disrupted by combined cold-salt stress, thereby inhibiting cold-responsive aggregation in high-salt conditions. ATP desorbs salt cations from negatively charged surfaces through predominant interactions between ATP and the salt cations. Although the mode of ATP's action remains unchanged with temperature, the extent of interaction scales with temperature, requiring less ATP activity at lower temperatures, justifying the reason for reduction in cellular ATP content due to the cold effect, reported in previous experimental studies. The trend observed in inorganic nanostructures is recurrent and robustly transferable to charged protein interfaces. A thorough comparison of ATP's cryoprotective activity with traditionally known biological cryoprotectants (glycine and betaine) reveals ATP's greater efficiency. In retrospect, our findings highlight ATP's additional biological role in cryopreservation, expanding its potential biomedical applications by offering effective protection of cells from cryoinjuries and avoiding the significant challenges associated with the toxicity of organic cryoprotectants.

Optical wavefront shaping in deep tissue using photoacoustic feedback

Abstract Over the past decade, optical wavefront shaping has been developed to focus light through highly opaque scattering layers, opening new possibilities for biomedical applications. To probe light intensity deep inside soft scattering media such as biological tissues, internal guide-stars are required. Here, we give an overview of the main principles and describe in depth the use of a photoacoustic feedback signal for this purpose. We further present first principles calculations and simulations to estimate important experimental parameters, and detailed instructions on designing and conducting these experiments. Finally, we provide guidance towards selecting suitable equipment for building a typical experimental setup, paving the way for further innovative biomedical imaging and therapy applications.

Engineered Shape-Morphing Transitions in Hydrogels Through Suspension Bath Printing of Temperature-Responsive Granular Hydrogel Inks.

4D printing of hydrogels is an emerging technology used to fabricate shape-morphing soft materials that are responsive to external stimuli for use in soft robotics and biomedical applications. Soft materials are technically challenging to process with current 4D printing methods, which limits the design and actuation potential of printed structures. Here, a simple multi-material 4D printing technique is developed that combines dynamic temperature-responsive granular hydrogel inks based on hyaluronic acid, whose actuation is modulated via poly(N-isopropylacrylamide) crosslinker design, with granular suspension bath printing that provides structural support during and after the printing process. Granular hydrogels are easily extruded upon jamming due to their shear-thinning properties and their porous structure enables rapid actuation kinetics (i.e., seconds). Granular suspension baths support responsive ink deposition into complex patterns due to shear-yielding to fabricate multi-material objects that can be post-crosslinked to obtain anisotropic shape transformations. Dynamic actuation is explored by varying printing patterns and bath shapes, achieving complex shape transformations such as 'S'-shaped and hemisphere structures. Furthermore, stepwise actuation is programmed into multi-material structures by using microgels with varied transition temperatures. Overall, this approach offers a simple method to fabricate programmable soft actuators with rapid kinetics and precise control over shape morphing.

Fe0-MAP Prepared Glycosurfaces for Selective Cell Capture: From Adherent to Suspended Cells.

The specific capture of live cells is crucial for various biomedical applications. Existing methods often are limited by complex production processes. This study introduces Fe0-mediated monomer-adaptation polymerization (Fe0-MAP), a convenient and rapid synthesis approach for selective cell capture using surface-engineered glycopolymer brushes. This method utilizes surface-initiated zerovalent iron-mediated reversible-deactivation radical polymerization (Fe0-SI-RDRP), offering advantages like simplicity, biocompatibility and oxygen-tolerance due to the use of iron sheet as catalysts. We successfully employed Fe0-MAP to selective capture both adherent (HeLa, L929) and suspended cells (Ramos, U937) in mammalian cell cultures. Combining excellent biocompatibility, specific and reusable cell capture capabilities, and applicability to suspended cells, Fe0-MAP establishes itself as a promising strategy for selective cell capture, holding significant potential for diverse biomedical applications.

Eco-friendly synthesis of bioactive silver nanoparticles from black roasted gram (Cicer arietinum) for biomedical applications

Green synthesis leverages biological resources such as plant extracts to produce cost-effectively and environmentally friendly NPs. In our study, silver nanoparticles (AgNPs) are biosynthesized using blank roasted grams (Cicer arietinum) as reducing agents. CA-AgNPs were characterized by a characteristic surface plasmon resonance (SPR) peak at 224 nm in the UV–Vis spectrum. FTIR analysis revealed functional groups with O–H stretching at 3410 cm−1, C–H stretching at 2922 cm−1, and C=O stretching at 1635 cm−1. XRD patterns exhibited sharp peaks at 33.2°, 38.4°, 55.7°, and 66.6°, confirming high crystallinity. Morphological analysis through FESEM indicated spherical CA-AgNPs averaging 500 nm in size, with EDS revealing Ag at 97.51% by weight. Antimicrobial assays showed zones of inhibition of 14 mm against Candida albicans, 18 mm against Escherichia coli., and 12 mm against Propionibacterium acnes. The total phenolic content of CA-AgNPs was 26.17 ± 13.54 mg GAE/g, significantly higher than the 11.85 ± 9.57 mg GAE/g in CA extract. The ABTS assay confirmed the antioxidant potential with a lower IC50 value of 1.73 ± 0.41 µg/mL, indicating enhanced radical scavenging activity. Anti-melanogenesis was validated through tyrosinase, showing inhibition rates of 97.97% at the highest concentrations. The anti-inflammatory was evaluated by western blot, which showed decreased expression of iNOS and COX-2. This study demonstrates the green synthesis of CA-AgNPs and its potential biomedical applications. The results of this study demonstrate that biosynthesized CA-AgNPs have key biological applications.

Natural Biological Solutions for Chronic Pathological Problems

Naturally sourced biomolecules and their derivatives have had significant historical impacts in terms of their biomedical application [...]

Nitrogen-doped carbon quantum dot regulates cell proliferation and differentiation by endoplasmic reticulum stress

ABSTRACT Quantum dots have diverse biomedical applications, from constructing biological infrastructures like medical imaging to advancing pharmaceutical research. However, concerns about human health arise due to the toxic potential of quantum dots based on heavy metals. Therefore, research on quantum dots has predominantly focused on oxidative stress, cell death, and other broader bodily toxicities. This study investigated the toxicity and cellular responses of mouse embryonic stem cells (mESCs) and mouse adult stem cells (mASCs) to nitrogen-doped carbon quantum dots (NCQDs) made of non-metallic materials. Cells were exposed to NCQDs, and we utilized a fluorescent ubiquitination-based cell system to verify whether NCQDs induce cytotoxicity. Furthermore, we validated the differentiation-inducing impact of NCQDs by utilizing embryonic stem cells equipped with the Oct4 enhancer-GFP reporter system. By analyzing gene expression including Crebzf, Chop, and ATF6, we also observed that NCQDs robustly elicited endoplasmic reticulum (ER) stress. We confirmed that NCQDs induced cytotoxicity and abnormal differentiation. Interestingly, we also confirmed that low concentrations of NCQDs stimulated cell proliferation in both mESCs and mASCs. In conclusion, NCQDs modulate cell death, proliferation, and differentiation in a concentration-dependent manner. Indiscriminate biological applications of NCQDs have the potential to cause cancer development by affecting normal cell division or to fail to induce normal differentiation by affecting embryonic development during pregnancy. Therefore, we propose that future biomedical applications of NCQDs necessitate comprehensive and diverse biological studies.

Living Cell-Mediated Catalyst-Free Spontaneous Polymerization of Zwitterionic Methacrylates for Preparation of Probiotic-Loaded Hydrogels.

Living cell-mediated polymerization offers promising applications in biomaterials, yet its further biological utilization is hindered by the need for metal ions or radical initiators with available methods. In this study, we introduce a living cell-mediated polymerization that leverages the intrinsic metabolic activities of living cells to initiate and sustain free radical polymerization of zwitterionic methacrylates. The polymerization proceeded in the absence of transition metal catalysts, radical initiators, or light sources. The conversion of zwitterionic methacrylate strongly correlated with cellular activities and achieved a maximum conversion of 98% within 48 hours. Living cells efflux redox power across membranes through metabolism and that terminal electron fluxes are captured by zwitterionic methacrylates pre-assembled on the living cell surface to initiate radical polymerization reactions. The polymerization caused significant changes to the cell membrane surface and synthesized hydrogels with tailored mechanical properties. The polymer hydrogel obtained via probiotic E. coli Nissle 1917 was able to release the in-situ encapsulated molecules, which demonstrated living cell-mediated polymer hydrogel as a vehicle for the delivery of both cellular and molecular therapeutic agents. This research offered a green and efficient method for synthesizing bioactive materials and advancing the field of cellular therapeutics and drug delivery.

Living Cell‐Mediated Catalyst‐Free Spontaneous Polymerization of Zwitterionic Methacrylates for Preparation of Probiotic‐Loaded Hydrogels

Living cell‐mediated polymerization offers promising applications in biomaterials, yet its further biological utilization is hindered by the need for metal ions or radical initiators with available methods. In this study, we introduce a living cell‐mediated polymerization that leverages the intrinsic metabolic activities of living cells to initiate and sustain free radical polymerization of zwitterionic methacrylates. The polymerization proceeded in the absence of transition metal catalysts, radical initiators, or light sources. The conversion of zwitterionic methacrylate strongly correlated with cellular activities and achieved a maximum conversion of 98% within 48 hours. Living cells efflux redox power across membranes through metabolism and that terminal electron fluxes are captured by zwitterionic methacrylates pre‐assembled on the living cell surface to initiate radical polymerization reactions. The polymerization caused significant changes to the cell membrane surface and synthesized hydrogels with tailored mechanical properties. The polymer hydrogel obtained via probiotic E. coli Nissle 1917 was able to release the in‐situ encapsulated molecules, which demonstrated living cell‐mediated polymer hydrogel as a vehicle for the delivery of both cellular and molecular therapeutic agents. This research offered a green and efficient method for synthesizing bioactive materials and advancing the field of cellular therapeutics and drug delivery.

Creation of 3D chitin/chitosan composite scaffold from naturally pre-structured verongiid sponge skeleton

This study represents the first creation and characterization of a 3D chitin/chitosan composite scaffold derived from the naturally pre-structured skeleton of the cultivated marine demosponge Aplysina aerophoba, aiming to preserve the intricate architecture of the unique tube-like chitin while incorporating chitosan layers. Advanced staining methods, including the use of iodine and Cibacron Brilliant Red (CBR), were employed to distinguish these polysaccharides. ATR-FTIR spectroscopy confirmed the system's structural integrity and identified the optimal chitin/chitosan balance, achieved after 60-minute treatment in 38% NaOH at 95°C. Fluorescent microscopy using fluorescein isothiocyanate (FITC) effectively confirmed the presence of chitosan layers in the created chitin/chitosan scaffolds. Scanning electron microscopy analysis further elucidated significant morphological distinctions, where chitin fibers displayed a smooth, uniform surface, contrasting with the ragged and irregular texture of chitosan-containing fibers, indicating significant surface modifications. Zeta potential measurements confirmed the partial transformation of chitin into chitosan. The dual-layer configuration, consisting of a resilient chitin core and a versatile chitosan exterior, not only provides structural support, but also enhances the scaffold's functionality for potential technological and biomedical applications. The preferential metallization of the chitosan phase by copper nanoparticles in the created 3D chitin/chitosan composite opens the way to the potential use of such scaffolds in catalysis.