J. Funct. Biomater., Volume 12, Issue 4 (December 2021) – 24 articles

The development of asymmetric membranes—i.e., matching two fibrous layers with selected composition and morphological properties to mimic both the epidermis and dermis—currently represents one of the most promising strategies to support skin regeneration during the wound healing process. Herein, a new asymmetric platform fabricated by a sequential electrospinning process was investigated. The top layer comprises cross-linked polyvinylalcohol (PVA) nanofibers (NFs)—from water solution—to replicate the epidermis’s chemical stability and wettability features. Otherwise, the bottom layer is fabricated by integrating PVA with wool-keratin extracted via sulfitolysis. This protein is a biocompatibility polymer with excellent properties for dermis-like structures. Morphological characterization via SEM supported by image analysis showed that the asymmetric membrane exhibited average fiber size—max frequency diameter 450 nm, range 1.40 μm—and porosity suitable for the healing process. FTIR-spectrums confirmed the presence of keratin in the bottom layer and variations of keratin-secondary structures. Compared with pure PVA-NFs, keratin/PVA-NFs showed a significant improvement in cell adhesion in in vitro tests. In perspective, these asymmetric membranes could be promisingly used to confine active species (i.e., antioxidants, antimicrobials) to the bottom layer to support specific cell activities (i.e., proliferation, differentiation) in wound healing applications. Full article

Breast cancer is becoming one of the main lethal carcinomas in the recent era, and its occurrence rate is increasing day by day. There are different breast cancer biomarkers, and their overexpression takes place in the metastasis of cancer cells. The most prevalent breast cancer biomarker is the human epidermal growth factor receptor2 (HER2). As this biomarker is overexpressed in malignant breast tissues, it has become the main focus in targeted therapies to fight breast cancer. There is a cascade of mechanisms involved in metastasis and cell proliferation in cancer cells. Nanotechnology has become extremely advanced in targeting and imaging cancerous cells. Quantum dots (QDs) are semiconductor NPs, and they are used for bioimaging, biolabeling, and biosensing. They are synthesized by different approaches such as top-down, bottom-up, and synthetic methods. Fully human monoclonal antibodies synthesized using transgenic mice having human immunoglobulin are used to target malignant cells. For the HER2 receptor, herceptin^® (trastuzumab) is the most specific antibody (Ab), and it is conjugated with QDs by using different types of coupling mechanisms. This quantum dot monoclonal antibody (QD-mAb) conjugate is localized by injecting it into the blood vessel. After the injection, it goes through a series of steps to reach the intracellular space, and bioimaging of specifically the HER2 receptor occurs, where apoptosis of the cancer cells takes place either by the liberation of Ab or the free radicals. Full article

Rutin is a natural compound with several pharmacological effects. Among these, antioxidant activity is one of the best known. Despite its numerous benefits, its topical application is severely limited by its physicochemical properties. For this reason, the use of suitable systems could be necessary to improve its delivery through skin, thus enhancing its pharmacological effects. In this regard, the aim of this work is to optimize the ethosomal dispersion modifying both lipid and ethanol concentrations and encapsulating different amounts of rutin. Characterization studies performed on the realized systems highlighted their great stability properties. Studies of encapsulation efficiency and loading degree allowed us to identify a better formulation (EE% 67.5 ± 5.2%, DL% 27 ± 1.7%), which was used for further analyses. The data recorded from in vitro studies showed that the encapsulation into these nanosystems allowed us to overcome the photosensitivity limitation of rutin. Indeed, a markable photostability of the loaded formulation was recorded, compared with that reported from the free rutin solution. The efficacy of the nanosystems was finally evaluated both in vitro on keratinocyte cells and in vivo on human healthy volunteers. The results confirmed the potentiality of rutin-loaded nanosystems for skin disease, mainly related to their anti-inflammatory and antioxidant effects. Full article

Enamel demineralization around orthodontic adhesive is a common esthetic concern during orthodontic treatment. The aim of this study was to prepare orthodontic adhesives containing monocalcium phosphate monohydrate (MCPM) and nisin to enable mineralizing and antibacterial actions. The physicomechanical properties and the inhibition of S. mutans growth of the adhesives with added MCPM (5, 10 wt %) and nisin (5, 10 wt %) were examined. Transbond XT (Trans) was used as the commercial comparison. The adhesive containing a low level of MCPM showed significantly higher monomer conversion (42–62%) than Trans (38%) (p < 0.05). Materials with additives showed lower monomer conversion (p < 0.05), biaxial flexural strength (p < 0.05), and shear bond strength to enamel than those of a control. Additives increased water sorption and solubility of the experimental materials. The addition of MCPM encouraged Ca and P ion release, and the precipitation of calcium phosphate at the bonding interface. The growth of S. mutans in all the groups was comparable (p > 0.05). In conclusion, experimental orthodontic adhesives with additives showed comparable conversion but lesser mechanical properties than the commercial material. The materials showed no antibacterial action, but exhibited ion release and calcium phosphate precipitation. These properties may promote remineralization of the demineralized enamel. Full article

The development of nanotechnology has allowed us to better exploit the potential of many natural compounds. However, the classic nanotechnology approach often uses both dangerous and environmentally harmful chemical compounds and drastic conditions for synthesis. Nevertheless, “green chemistry” techniques are revolutionizing the possibility of making technology, also for tissue engineering, environmentally friendly and cost-effective. Among the many approaches proposed and among several natural compounds proposed, honey seems to be a very promising way to realize this new “green” approach. Full article

Polylactide acid (PLA), as an FDA-approved biomaterial, has been widely applied due to its unique merits, such as its biocompatibility, biodegradability, and piezoelectricity. Numerous utilizations, including sensors, actuators, and bio-application—its most exciting application to promote cell migration, differentiation, growth, and protein–surface interaction—originate from the piezoelectricity effect. Since PLA exhibits piezoelectricity in both crystalline structure and an amorphous state, it is crucial to study it closely to understand the source of such a phenomenon. In this respect, in the current study, we first reviewed the methods promoting piezoelectricity. The present work is a comprehensive review that was conducted to promote the low piezoelectric constant of PLA in numerous procedures. In this respect, its chemistry and structural origins have been explored in detail. Combining any other variables to induce a specific application or to improve any PLA barriers, namely, its hydrophobicity, poor electrical conductivity, or the tuning of its mechanical properties, especially in the application of cardiovascular tissue engineering, is also discussed wherever relevant. Full article

Gold nanoparticles (AuNPs) are extremely promising objects for solving a wide range of biomedical problems. The gold nanoparticles production by biological method (“green synthesis”) is eco-friendly and allows minimization of the amount of harmful chemical and toxic byproducts. This review is devoted to the AuNPs biosynthesis peculiarities using various living organisms (bacteria, fungi, algae, and plants). The participation of various biomolecules in the AuNPs synthesis and the influence of size, shapes, and capping agents on the functionalities are described. The proposed action mechanisms on target cells are highlighted. The biological activities of “green” AuNPs (antimicrobial, anticancer, antiviral, etc.) and the possibilities of their further biomedical application are also discussed. Full article

The meniscus is a critical component of a healthy knee joint. It is a complex and vital fibrocartilaginous tissue that maintains appropriate biomechanics. Injuries of the meniscus, particularly in the inner region, rarely heal and usually progress into structural breakdown, followed by meniscus deterioration and initiation of osteoarthritis. Conventional therapies range from conservative treatment, to partial meniscectomy and even meniscus transplantation. All the above have high long-term failure rates, with recurrence of symptoms. This communication presents a brief account of in vitro and in vivo studies and describes recent developments in the field of 3D-printed scaffolds for meniscus tissue engineering. Current research in meniscal tissue engineering tries to combine polymeric biomaterials, cell-based therapy, growth factors, and 3D-printed scaffolds to promote the healing of meniscal defects. Today, 3D-printing technology represents a big opportunity in the orthopaedic world to create more specific implants, enabling the rapid production of meniscal scaffolds and changing the way that orthopaedic surgeons plan procedures. In the future, 3D-printed meniscal scaffolds are likely to be available and will also be suitable substitutes in clinical applications, in an attempt to imitate the complexity of the native meniscus. Full article

Pancreatic β-cell loss and failure with subsequent deficiency of insulin production is the hallmark of type 1 diabetes (T1D) and late-stage type 2 diabetes (T2D). Despite the availability of parental insulin, serious complications of both types are profound and endemic. One approach to therapy and a potential cure is the immunoisolation of β cells via artificial cell microencapsulation (ACM), with ongoing promising results in human and animal studies that do not depend on immunosuppressive regimens. However, significant challenges remain in the formulation and delivery platforms and potential immunogenicity issues. Additionally, the level of impact on key metabolic and disease biomarkers and long-term benefits from human and animal studies stemming from the encapsulation and delivery of these cells is a subject of continuing debate. The purpose of this review is to summarise key advances in this field of islet transplantation using ACM and to explore future strategies, limitations, and hurdles as well as upcoming developments utilising bioengineering and current clinical trials. Full article

To match the current life-style, there is a huge demand and market for the processed food whose manufacturing requires multiple steps. The mounting demand increases the pressure on the producers and the regulatory bodies to provide sensitive, facile, and cost-effective methods to safeguard consumers’ health. In the multistep process of food processing, there are several chances that the food-spoiling microbes or contaminants could enter the supply chain. In this contest, there is a dire necessity to comprehend, implement, and monitor the levels of contaminants by utilizing various available methods, such as single-cell droplet microfluidic system, DNA biosensor, nanobiosensor, smartphone-based biosensor, aptasensor, and DNA microarray-based methods. The current review focuses on the advancements in these methods for the detection of food-borne contaminants and pathogens. Full article

The ideal positioning of immediate implants in molar extraction sockets often requires the osteotomy to be in the interradicular septum, which can be challenging in some cases, with traditional site preparation techniques. Patients who had undergone molar tooth extraction and immediate implant placement at five different centers, and followed up between August 2015 and September 2020, were evaluated. Inclusion criteria were use of the osseodensification technique for implant site preparation. The primary outcome was septum width measurement pre-instrumentation and osteotomy diameter post expansion. Clinical outcomes, such as implant insertion torque (ISQ) and implant survival rate, were also collected. A total of 131 patients, who received 145 immediate implants, were included. The mean overall septum width at baseline was 3.3 mm and the mean osteotomy diameter post instrumentation was 4.65 mm. A total of ten implants failed: seven within the healing period and three after loading; resulting in a cumulative implant survival rate of 93.1%. This retrospective study showed that osseodensification is a predictable method for immediate implant placement with interradicular septum expansion in molar extraction sockets. Furthermore, it allowed the introduction of a new molar socket classification. In the future, well-designed controlled clinical studies are needed to confirm these results and further explore the potential advantages of this technique. Full article

The effect of thermohydrogen treatment and vacuum ion–plasma nitriding on the determination of the volume and surface structure of ball heads made of Ti–6Al–4V alloy was studied. It was found that the submicrocrystalline structure formed in the head during thermohydrogen treatment makes it possible to achieve hardness values of 39–41 units HRC and a surface roughness of 0.02 μm. It was shown that the creation of a modified layer consisting of ε (TiN) and δ (Ti₂N) titanium nitrides on the surface of a ball head and the solid interstitial solution of nitrogen in α-titanium makes it possible to completely eliminate material wear when testing for friction on ultra-high-molecular-weight polyethylene. The equivalent analysis was also conducted with a ball head that had been implanted in a human body for 12 years. It was found that the change in the color of the head, from slightly golden after nitriding to metallic, is due to the formation of an oxynitride nanoscale layer on the surface. It was shown that in contrast with films made of titanium oxide, the film developed in this study has high wear resistance. Full article

Transplantation of differentiated and fully functional neurons may be a better therapeutic option for the cure of neurodegenerative disorders and brain injuries than direct grafting of neural stem cells (NSCs) that are potentially tumorigenic. However, the differentiation of NSCs into a large population of neurons has been a challenge. Nanomaterials have been widely used as substrates to manipulate cell behavior due to their nano-size, excellent physicochemical properties, ease of synthesis, and versatility in surface functionalization. Nanomaterial-based scaffolds and synthetic polymers have been fabricated with topology resembling the micro-environment of the extracellular matrix. Nanocellulose materials are gaining attention because of their availability, biocompatibility, biodegradability and bioactivity, and affordable cost. We evaluated the role of nanocellulose with different linkage and surface features in promoting neuronal differentiation. Nanocellulose coupled with lysine molecules (CNC–Lys) provided positive charges that helped the cells to attach. Embryonic rat NSCs were differentiated on the CNC–Lys surface for up to three weeks. By the end of the three weeks of in vitro culture, 87% of the cells had attached to the CNC–Lys surface and more than half of the NSCs had differentiated into functional neurons, expressing endogenous glutamate, generating electrical activity and action potentials recorded by the multi-electrode array. Full article

Improvements in software for image analysis have enabled advances in both medical and engineering industries, including the use of medical analysis tools to recreate internal parts of the human body accurately. A research analysis found that FDM-sourced elements have shown viability for a customized and reliable approach in the orthopedics field. Three-dimensional printing has allowed enhanced accuracy of preoperative planning, leading to reduced surgery times, fewer unnecessary tissue perforations, and fewer healing complications. Furthermore, using custom tools chosen for each procedure has shown the best results. Bone correction-related surgeries require customized cutting guides for a greater outcome. This study aims to assess the biopolymer-based tools for surgical operations and their ability to sustain a regular heat-sterilization cycle without compromising the geometry and fit characteristics for a proper procedure. To achieve this, a DICOM and FDM methodology is proposed for fast prototyping of the cutting guide by means of 3D engineering. A sterilization test was performed on HTPLA, PLA, and nylon polymers. As a result, the unique characteristics within the regular autoclave sterilization process allowed regular supplied PLA to show there were no significant deformations, whilst annealed HTPLA proved this material’s capability of sustaining repeated heat cycles due to its crystallization properties. Both of these proved that the sterilization procedures do not compromise the reliability of the part, nor the safety of the procedure. Therefore, prototypes made with a similar process as this proposal could be safely used in actual surgery practices, while nylon performed poorly because of its hygroscopic properties. Full article

Zirconia has been used as a prosthesis material for over a decade because of its excellent mechanical properties and esthetics. The surface treatment for zirconia generally involves sandblasting and the application of primers for favorable bond strength between the surface and resin. However, sandblasting causes the microcracking and chipping of the zirconia surface. To overcome these challenges, the metallization of the zirconia surface was performed. Ti and Au were sputtered on yttria stabilized zirconia (YSZ) disks and heated to 800 °C for 15 min in air. These disks were bonded to stainless-steel rods using resin cement. Then, shear bond strength tests were performed using an Instron-type testing machine. The shear bond strength of the Ti sputtering group was significantly higher than that of the other groups. According to the results of X-ray photoelectron spectroscopy and electron probe microanalysis, the Ti-sputtered YSZ surface contained both sub-titanium oxide and titanium oxide before heating. Sub-titanium oxide was converted to titanium oxide by heating. These results suggest that metallization using Ti is effective for zirconia surface treatment to improve the shear bond strength between YSZ and resin cement. This metallization technique for YSZ has potential in clinical applications. Full article

Chitosan/PVA hydrogel films crosslinked by the freeze–thaw method and containing honey and allantoin were prepared for application as wound dressing materials. The effects of the freeze–thaw process and the addition of honey and allantoin on the swelling, the gel content and the mechanical properties of the samples were evaluated. The physicochemical properties of the samples, with and without the freeze–thaw process, were compared using FTIR, DSC and XRD. The results showed that the freeze–thaw process can increase the crystallinity and thermal stability of chitosan/PVA films. The freeze–thaw process increased the gel content but did not have a significant effect on the tensile strength. The presence of honey reduced the swelling and the tensile strength of the hydrogels due to hydrogen bonding interactions with PVA and chitosan chains. Long-term cell culture experiments using normal human dermal fibroblast (NHDF) cells showed that the hydrogels maintained their biocompatibility, and the cells showed extended morphology on the surface of the hydrogels for more than 30 days. The presence of honey significantly increased the biocompatibility of the hydrogels. The release of allantoin from the hydrogel was studied and, according to the Korsmeyer–Peppas and Weibull models, the mechanism was mainly diffusional. The results for the antimicrobial activity against E. coli and S. aureus bacteria showed that the allantoin-containing samples had a more remarkable antibacterial activity against S. aureus. According to the wound healing experiments, 98% of the wound area treated by the chitosan/PVA/honey hydrogel was closed, compared to 89% for the control. The results of this study suggest that the freeze–thaw process is a non-toxic crosslinking method for the preparation of chitosan/PVA hydrogels with long term biocompatibility that can be applied for wound healing and skin tissue engineering. Full article

In this study, we assessed the outcomes after surgical treatment of thoracic post-excision defects in 15 patients, using TiNi knitted surgical meshes and customized artificial TiNi-based ribs. Methods: Eight patients were diagnosed with advanced non-small cell lung cancer (NSCLC) invading the chest wall, of which five patients were T₃N₀M₀, two were T₃N₁M₀, and one was T₃N₂M₀. Squamous cell carcinoma was identified in three of these patients and adenocarcinoma in five. In two cases, chest wall resection and repair were performed for metastases of kidney cancer after radical nephrectomy. Three-dimensional CT reconstruction and X-ray scans were used to plan the surgery and customize the reinforcing TiNi-based implants. All patients received TiNi-based devices and were prospectively followed for a few years. Results: So far, there have been no lethal outcomes, and all implanted devices were consistent in follow-up examinations. Immediate complications were noted in three cases (ejection of air through the pleural drains, paroxysm of atrial fibrillation, and pleuritis), which were conservatively managed. In the long term, no complications, aftereffects, or instability of the thoracic cage were observed. Conclusion: TiNi-based devices used for extensive thoracic lesion repair in this context are promising and reliable biomaterials that demonstrate good functional, clinical, and cosmetic outcomes. Full article

The rise of antibiotic resistance has become a major threat to human health and it is spreading globally. It can cause common infectious diseases to be difficult to treat and leads to higher medical costs and increased mortality. Hence, multifunctional polymeric nanofibers with distinctive structures and unique physiochemical properties have emerged as a neo-tool to target biofilm and overcome deadly bacterial infections. This review emphasizes electrospun nanofibers’ design criteria and properties that can be utilized to enhance their therapeutic activity for antimicrobial therapy. Also, we present recent progress in designing the surface functionalization of antimicrobial nanofibers with non-antibiotic agents for effective antibacterial therapy. Lastly, we discuss the future trends and remaining challenges for polymeric nanofibers. Full article

Polymers have always played a critical role in the development of novel drug delivery systems by providing the sustained, controlled and targeted release of both hydrophobic and hydrophilic drugs. Among the different polymers, polyamides or poly(amino acid)s exhibit distinct features such as good biocompatibility, slow degradability and flexible physicochemical modification. The degradation rates of poly(amino acid)s are influenced by the hydrophilicity of the amino acids that make up the polymer. Poly(amino acid)s are extensively used in the formulation of chemotherapeutics to achieve selective delivery for an appropriate duration of time in order to lessen the drug-related side effects and increase the anti-tumor efficacy. This review highlights various poly(amino acid) polymers used in drug delivery along with new developments in their utility. A thorough discussion on anticancer agents incorporated into poly(amino acid) micellar systems that are under clinical evaluation is included. Full article

Biological hydrogels are highly promising materials for bone tissue engineering (BTE) due to their high biocompatibility and biomimetic characteristics. However, for advanced and customized BTE, precise tools for material stabilization and tuning material properties are desired while optimal mineralisation must be ensured. Therefore, reagent-free crosslinking techniques such as high energy electron beam treatment promise effective material modifications without formation of cytotoxic by-products. In the case of the hydrogel gelatin, electron beam crosslinking further induces thermal stability enabling biomedical application at physiological temperatures. In the case of enzymatic mineralisation, induced by Alkaline Phosphatase (ALP) and mediated by Calcium Glycerophosphate (CaGP), it is necessary to investigate if electron beam treatment before mineralisation has an influence on the enzymatic activity and thus affects the mineralisation process. The presented study investigates electron beam-treated gelatin hydrogels with previously incorporated ALP and successive mineralisation via incubation in a medium containing CaGP. It could be shown that electron beam treatment optimally maintains enzymatic activity of ALP which allows mineralisation. Furthermore, the precise tuning of material properties such as increasing compressive modulus is possible. This study characterizes the mineralised hydrogels in terms of mineral formation and demonstrates the formation of CaP in dependence of ALP concentration and electron dose. Furthermore, investigations of uniaxial compression stability indicate increased compression moduli for mineralised electron beam-treated gelatin hydrogels. In summary, electron beam-treated mineralized gelatin hydrogels reveal good cytocompatibility for MG-63 osteoblast like cells indicating a high potential for BTE applications. Full article

Demineralized bone matrix (DBM) is a decalcified allo/xenograft retaining collagen and noncollagenous proteins, which has been extensively used because of its osteoconductive and osteoinductive properties. Calcium sulfate (CaSO₄, CS) is a synthetic bone substitute used in bone healing with biocompatible, nontoxic, bioabsorbable, osteoconductive, and good mechanical characteristics. This study aims to prepare a DBM/CS composite bone graft material in a moldable putty form without compromising the peculiar properties of DBM and CS. For this purpose, firstly, porcine femur was defatted using chloroform/methanol and extracted by acid for demineralization, then freeze-dried and milled/sieved to obtain DBM powder. Secondly, the α-form and β-form of calcium sulfate hemihydrate (CaSO₄·0.5H₂O, CSH) were produced by heating gypsum (CaSO₄·2H₂O). The morphology and particle sizes of α- and β-CSH were obtained by SEM, and their chemical properties were confirmed by EDS, FTIR and XRD. Furthermore, the DBM-based graft was mixed with α- or β-CSH at a ratio of 9:1, and glycerol/4% HPMC was added as a carrier to produce a putty. DBM/CSH putty possesses a low washout rate, good mechanical strength and biocompatibility. In conclusion, we believe that the moldable DBM/CSH composite putty developed in this study could be a promising substitute for the currently available bone grafts, and might have practical application in the orthopedics field as a potential bone void filler. Full article

The creation of artificial biocomposites consisting of biocompatible materials in combination with bioactive molecules is one of the main tasks of tissue engineering. The development of new materials, which are biocompatible, functional, and also biodegradable in vivo, is a specific problem. Two types of products can be formed from these materials in the processes of biodegradation. The first types of substances are natural for a living organism and are included in the metabolism of cells, for example, sugars, lactic, glycolic, and β-hydroxybutyric acids. Substances that are not metabolized by cells represent the other type. In the latter case, such products should not be toxic, and their concentration when entering the bloodstream should not exceed the established maximum permissible level. The composite materials based on a mixture of biodegradable synthetic and natural polymers with the addition of hydroxyapatite nanoparticles, which acts as a stabilizer of the dispersed system during production of the composite, and which is a biologically active component of the resulting matrix, were obtained and studied. The indirect effect of the shape, size, and surface charge of hydroxyapatite nanoparticles on the structure and porosity of the formed matrix was shown. An in vivo study showed the absence of acute toxicity of the developed composites. Full article

Gestational diabetes mellitus (GDM) is the most frequent complication during pregnancy. This complex disease is characterized by glucose intolerance and consequent hyperglycemia that begins or is first diagnosed in pregnancy, and affects almost 7% of pregnant women. Previous reports have shown that GDM is associated with increased pregnancy complications and might cause abnormal fetal development. At present, treatments are not suitable for the prevention and management of these patients. As an alternative therapeutic opportunity and a leading scientific technique, nanotechnology has helped enlighten the health of these affected women. Theranostic nanomaterials with unique properties and small sizes (at least <100 nm in one of their dimensions) have been recently engineered for clinics and pharmaceutics. Reducing materials to the nanoscale has successfully changed their properties and enabled them to uniquely interact with cell biomolecules. Several biosensing methods have been developed to monitor glucose levels in GDM patients. Moreover, cerium oxide nanoparticles (NPs), selenium NPs, polymeric NPs, and drug-loaded NPs loaded with therapeutic agents have been used for GDM treatment. Still, there are some challenges associated with the detection limits and toxicity of such nanomaterials. This preliminary review covers the aspects from a fast-developing field to generating nanomaterials and their applications in GDM diagnosis and treatment. Full article

A copper oxide/cerium oxide nanocomposite (CuO/CeO₂, NC) was synthesized via a novel method using a metal–organic framework as a precursor. This nanomaterial was characterized by Fourier transform infrared spectroscopy (FTIR), powder X-ray diffraction (PXRD), field emission scanning electron microscopy (FESEM), transmission electron microscopy (TEM), dynamic light scattering size analysis (DLS), and zeta potential. The PXRD showed the successful synthesis of the CuO/CeO₂ NC, in which the 2theta values of 35.55° (d = 2.52 Å, 100%) and 38.73° (d = 2.32 Å, 96%) revealed the existence of copper (II) oxide. FTIR analysis showed the CeO₂, hydroxyl groups, absorbed water, and some residual peaks. The solid phase analysis by FESEM and TEM images showed mean particle sizes of 49.18 ± 24.50 nm and 30.58 ± 26.40 nm, respectively, which were comparable with crystallite size (38.4 nm) obtained from PXRD, but it appears the CuO/CeO₂ NC was not evenly distributed and in some areas, showed it was highly agglomerated. The hydrodynamic size (750.5 nm) also showed the agglomeration of the CuO/CeO₂ NCs in the solution, which had a negatively charged surface. The CuO/CeO₂ NCs showed anti-proliferative activity against human breast cancer cell line (MCF-7) in a dose- and time-dependence way, while affecting normal cells less significantly. Full article