LSPR sensing for in situ monitoring the Ag dissolution of Au@Ag core-shell nanoparticles in biological environments.

Silver nanoparticles (AgNPs) are extensively used as an antibacterial agent, and monitoring the dissolution behavior of AgNPs in native biological environments is critical in both optimizing their performance and regulating their safety. However, current assessment methods rely on sophisticated analytical tools that are off-site and time-consuming with potential underestimations, due to complicated sample preparation. Although localized surface plasmon resonance (LSPR) sensing offers a facile method for the detection of AgNP dissolution, it is limited by low sensitivity and poor nanoparticle stability in native biological environments. Herein, we constructed a highly sensitive and stable LSPR sensor using gold-silver core-shell nanoparticles (Au@AgNPs), in combination with polymeric stabilizing agents, for the direct measurement of the Ag shell dissolution in native biological media. The high sensitivity was attributed to the acute and large LSPR shift generated by bimetallic nanoparticles. The sensor was used for the real-time monitoring of the Ag dissolution of Au@AgNPs during their co-culture with both bacteria and fibroblast cells. The media pH was found to dominate the Ag dissolution process, where Au@AgNPs exhibited bactericidal effects in the bacteria environment with relatively low pH, but they showed little toxicity towards fibroblast cells at pH 7.4. The minimum inhibition concentration of Au@AgNPs for bacterial growth was found similar to that of AgNO3 in terms of released Ag amount. Thus, stabilized Au@AgNPs not only allow the in-situ monitoring of Ag dissolution via LSPR sensing but also constitute an effective antibacterial agent with controlled toxicity, holding great potential for future biomedical and healthcare applications.

Autoclavable Albumin-Based Cryogels with Uncompromising Properties.

The development of autoclavable hydrogels has been driven by the need for materials that can withstand the rigors of sterilization without compromising their properties or functionality. Many conventional hydrogels cannot withstand autoclave treatment owing to the breakdown of their composition or structure under the high-temperature and high-pressure environment of autoclaving. Here, the effect of autoclaving on the physical, mechanical, and biological properties of bovine serum albumin methacryloyl (BSAMA) cryogels at three protein concentrations (3, 5, and 10%) was extensively studied. We found that BSAMA cryogels at three concentrations remained little changed after autoclaving in terms of gross shape, pore structure, and protein secondary structure. Young's modulus of autoclaved BSAMA cryogels (BSAMAA) at low concentrations (3 and 5%) was similar to that of BSAMA cryogels, whereas 10% BSAMAA exhibited a higher Young's modulus value, compared with 10% BSAMA. Interestingly, BSAMAA cryogels prolonged degradation. Importantly, cell viability, drug release, and hemolytic behaviors were found to be similar among the pre- and post-autoclaved cryogels. Above all, autoclaving proved to be more effective in sterilizing BSAMA cryogels from bacteria contamination than UV and ethanol treatments. Thus, autoclavable BSAMA cryogels with uncompromising properties would be useful for biomedical applications.

Emerging albumin hydrogels as personalized biomaterials.

Developing biomaterials-based tissue engineering scaffolds with personalized features and intrinsic biocompatibility is appealing and urgent. Through utilizing various strategies, albumin, as the most abundant protein in plasma, could be fabricated into sustainable, cost-effective, and potentially personalized hydrogels that would display enormous biological applications. To date, much of the albumin-based research is primarily engrossed in using albumin as a therapeutic molecule or a drug carrier, not much as a scaffold for tissue engineering. For this reason, we have come up with a detailed and insightful review of recent progress in albumin-based hydrogels having an emphasis on production techniques, material characteristics, and biological uses. It is envisioned that albumin-based scaffolds would be appealing and useful platforms to meet current tissue engineering needs and achieve the goal of clinical translation to benefit patients. STATEMENT OF SIGNIFICANCE: The creation of autologous material-based scaffolds is a potential method for preventing immunological reactions and obtaining the best therapeutic results. Patient-derived albumin hydrogels may consequently provide improved opportunities for personalized treatment due to their abundant supply and minimal immunogenicity. To provide a detailed and insightful summary on albumin-based hydrogels, this review includes latest comprehensive information on their preparation procedures, features, and applications in 3D printing and other biomedical applications. The challenges, along with the future potential for implementing albumin-based hydrogels in clinics, have also been addressed.

Human Albumin-Based Hydrogels for Their Potential Xeno-Free Microneedle Applications.

Nowadays, hydrogels-based microneedles (MNs) have attracted a great interest owing to their outstanding qualities for biomedical applications. For the fabrication of hydrogels-based microneedles as tissue engineering scaffolds and drug delivery carriers, various biomaterials have been tested. They are required to feature tunable physiochemical properties, biodegradability, biocompatibility, nonimmunogenicity, high drug loading capacity, and sustained drug release. Among biomaterials, human proteins are the most ideal biomaterials for fabrication of hydrogels-based MNs; however, they are mechanically weak and poorly processible. To the best of the knowledge, there are no reports of xeno-free human protein-based MNs so far. Here, human albumin-based hydrogels and microneedles for tissue engineering and drug delivery by using relatively new processible human serum albumin methacryloyl (HSAMA) are engineered. The resultant HSAMA hydrogels display tunable mechanical properties, biodegradability, and good biocompatibility. Moreover, the xeno-free HSAMA microneedles display a sustained drug release profile and significant mechanical strength to penetrate the model skin. In vitro, they also show good biocompatibility and anticancer efficacy. Sustainable processible human albumin-based biomaterials may be employed as a xeno-free platform in vivo for tissue engineering and drug delivery in clinical trials in the future.

Mussel-Inspired Surface Functionalization of Porous Albumin Cryogels Supporting Synergistic Antibacterial/Antioxidant Activity and Bone-Like Apatite Formation.

A crucial method for adding new functions to current biomaterials for biomedical applications has been surface functionalization via molecular design. Mussel-inspired polydopamine (PDA) has generated much attention as a facile method for the functionalization of biomaterials because of its substantial independence in deposition, beneficial cell interactions, and significant responsiveness aimed at secondary functionalization. Because of their porous structure, the bovine serum albumin methacryloyl (BSAMA)-BM cryogels were functionalized with PDA (BM-PDA), which may reproduce the architecture and biological purpose of the natural extracellular environment. Excellent antioxidative and antibacterial qualities, improved mineralization, and better cell responsiveness were all demonstrated by BM-PDA. BM-PDA scaffolds maintained their linked and uniform pores after functionalization, which can make it easier for nutrients to be transported during bone repair. As a result, hydroxyapatite (HA)-coated BM* and BM-PDA* cryogels were created through successive mineralization with the goal of mineralized bone tissue repair. The heterogeneous nucleation and surface roughness contributed to rod-like apatite production in BM-PDA* cryogels whereas BM* cryogels were made up of plate-like HA morphologies. Analysis results showed that after five cycles, the mineral contents were around 57% and the HA units remained equally dispersed on the surface of BM-PDA* with a Ca/P ratio of 1.63. Other natural polymer-based cryogels can be coated using this general, rapid, and simple PDA coating technique and utilized as implants for bone tissue engineering. Future clinical uses of albumin cryogels for bone tissue engineering will advance as a result of additional in-vivo testing of such PDA-coated cryogels.

An Engineered Protein-Based Building Block (Albumin Methacryloyl) for Fabrication of a 3D In Vitro Cryogel Model.

Drug-induced liver injury (DILI) is a leading cause of attrition in drug development or withdrawal; current animal experiments and traditional 2D cell culture systems fail to precisely predict the liver toxicity of drug candidates. Hence, there is an urgent need for an alternative in vitro model that can mimic the liver microenvironments and accurately detect human-specific drug hepatotoxicity. Here, for the first time we propose the fabrication of an albumin methacryloyl cryogel platform inspired by the liver's microarchitecture via emulating the mechanical properties and extracellular matrix (ECM) cues of liver. Engineered crosslinkable albumin methacryloyl is used as a protein-based building block for fabrication of albumin cryogel in vitro models that can have potential applications in 3D cell culture and drug screening. In this work, protein modification, cryogelation, and liver ECM coating were employed to engineer highly porous three-dimensional cryogels with high interconnectivity, liver-like stiffness, and liver ECM as artificial liver constructs. The resulting albumin-based cryogel in vitro model provided improved cell-cell and cell-material interactions and consequently displayed excellent liver functional gene expression, being conducive to detection of fialuridine (FIAU) hepatotoxicity.

Facile Fabrication of Transparent and Opaque Albumin Methacryloyl Gels with Highly Improved Mechanical Properties and Controlled Pore Structures.

For porous protein scaffolds to be employed in tissue-engineered structures, the development of cost-effective, macroporous, and mechanically improved protein-based hydrogels, without compromising the original properties of native protein, is crucial. Here, we introduced a facile method of albumin methacryloyl transparent hydrogels and opaque cryogels with adjustable porosity and improved mechanical characteristics via controlling polymerization temperatures (room temperature and -80 °C). The structural, morphological, mechanical, and physical characteristics of both porous albumin methacryloyl biomaterials were investigated using FTIR, CD, SEM, XRD, compression tests, TGA, and swelling behavior. The biodegradation and biocompatibility of the various gels were also carefully examined. Albumin methacryloyl opaque cryogels outperformed their counterpart transparent hydrogels in terms of mechanical characteristics and interconnecting macropores. Both materials demonstrated high mineralization potential as well as good cell compatibility. The solvation and phase separation owing to ice crystal formation during polymerization are attributed to the transparency of hydrogels and opacity of cryogels, respectively, suggesting that two fully protein-based hydrogels could be used as visible detectors/sensors in medical devices or bone regeneration scaffolds in the future.

Novel biohybrid spongy scaffolds for fabrication of suturable intraoral graft substitutes.

Despite the fact that classic autograft is the gold standard material for periodontal plastic surgery, it has some drawbacks, including the need for a second surgical site and the scarcity of palatal donor tissues. However, only a few research works on the manufacturing of bioengineered intraoral connective tissue grafts have been conducted. In this work, porous bovine serum albumin methacryloyl/gelatin methacryloyl (BG) biohybrid scaffolds were developed for super-elasticity, shape recovery, suturability for persistent stability, sufficient scaffolding function, and convenient manipulating characteristics to fabricate an intraoral graft substitute with superb stability to resist frequent dynamic forces caused by functional movement (speaking, masticating, and swallowing). Furthermore, in a 3D cell culture assay, BG scaffolds demonstrated excellent cell adhesion and proliferation of L929 cells. In addition, the BG scaffolds were able to release Ibuprofen in a controlled manner for postoperative recovery. The use of a low-cost, optimized cryogelation technique for functional biomacromolecules offers up new possibilities to develop promising scaffolds for dental clinical settings.

Synthesis and fabrication of gelatin-based elastomeric hydrogels through cosolvent-induced polymer restructuring.

Hydrogels have a wide range of applications in tissue engineering, drug delivery, device fabrication for biological studies and stretchable electronics. For biomedical applications, natural polymeric hydrogels have general advantages such as biodegradability and non-toxic by products as well as biocompatibility. However, applications of nature derived hydrogels have been severely limited by their poor mechanical properties. For example, most of the protein derived hydrogels do not exhibit high stretchability like methacrylated gelatin hydrogel has ∼11% failure strain when stretched. Moreover, protein derived elastomeric hydrogels that are fabricated from low molecular weight synthetic peptides require a laborious process of synthesis and purification. Biopolymers like gelatin, produced in bulk for pharma and the food industry can provide an alternative for the development of elastomeric hydrogels. Here, we report the synthesis of ureidopyrimidinone (Upy) functionalized gelatin and its fabrication into soft elastomeric hydrogels through supramolecular interactions that could exhibit high failure strain (318.73 ± 44.35%). The hydrogels were fabricated through a novel method involving co-solvent optimization and structural transformation with 70% water content. It is anticipated that the hydrogel fabrication method involves the formation of hydrophobic cores of ureidopyrimidinone groups inside the hydrogel which introduced elastomeric properties to the resulting hydrogel.

Comparison of globular albumin methacryloyl and random-coil gelatin methacryloyl: Preparation, hydrogel properties, cell behaviors, and mineralization.

Bovine serum albumin methacryloyl (BSAMA) is a newly emerging photocurable globular protein-based material whereas gelatin methacryloyl (GelMA) is one of the most popular photocurable fibrous protein-based materials. So far, the influence of their different structural conformations as building blocks on hydrogel properties and mineral deposition has not been investigated. Here, we compared their differences in structures, gelation kinetics, hydrogel properties, mineralization, and cell behaviors. BSAMA maintained a stable globular structure while GelMA exhibited temperature-sensitive conformations (4 - 37 °C). BSAMA displayed slower gelation kinetics and much more retarded enzymatic degradation compared to GelMA. Photocurable BSAMA (6.41 - 390.95 kPa) and GelMA hydrogels (36.09 - 199.70 kPa) exhibited tunable mechanical properties depending on their concentrations (10 - 20%). Interestingly, BSAMA hydrogels mineralized needle-like apatite (Ca/P: 1.409) with higher crystallinity compared to GelMA hydrogels (Ca/P: 1.344). BSAMA and GelMA supported satisfactory cell (MC3T3-L1) viability of 99.43 ± 0.57% and 97.14 ± 0.69%, respectively. However, BSAMA gels were less favorable to cell proliferation and migration than GelMA gels. In serum-free environments, cells on GelMA displayed a higher amount of attachment, a more elongated shape, and a longer protrusion compared to those on BSAMA (p < 0.01) during the early adhesion.

Recent advances in bioprinting technologies for engineering hepatic tissue.

In the sphere of liver tissue engineering (LTE), 3D bioprinting has emerged as an effective technology to mimic the complex in vivo hepatic microenvironment, enabling the development of functional 3D constructs with potential application in the healthcare and diagnostic sector. This review gears off with a note on the liver's microscopic 3D architecture and pathologies linked to liver injury. The write-up is then directed towards unmasking recent advancements and prospects of bioprinting for recapitulating 3D hepatic structure and function. The article further introduces available stem cell opportunities and different strategies for their directed differentiation towards various hepatic stem cell types, including hepatocytes, hepatic sinusoidal endothelial cells, stellate cells, and Kupffer cells. Another thrust of the article is on understanding the dynamic interplay of different hepatic cells with various microenvironmental cues, which is crucial for controlling differentiation, maturation, and maintenance of functional hepatic cell phenotype. On a concluding note, various critical issues and future research direction towards clinical translation of bioprinted hepatic constructs are discussed.

Pristane-induced mammary carcinomas.

Animal models for studying human breast cancer carcinogenesis and testing drug candidates on human breast cancer have extensively been proposed. Especially, chemically induced breast tumor models have been used because they can mimic the progression of clinical cancer from the beginning and can be generated with a facile procedure. Pristane is a hydrocarbon oil that is used as a chemical carcinogen to induce tumorigenesis in mice as well as arthritis and lupus nephritis in rats. In only a few studies, pristane-induced breast cancer models have been reported. This chapter is designated to describe pristane-induced mammary carcinoma models. Here, we provide a protocol for generating pristane-induced breast tumors in mice models for analyzing and for testing potential therapeutics on them. The given protocol can be applied to other animal models with some changes.

Hydrogel composite scaffolds with an attenuated immunogenicity component for bone tissue engineering applications.

Xenogeneic bones are potential templates for bone regeneration. In this study, decellularized porcine bone powder with attenuated immunogenicity was incorporated into a photocurable hydrogel, gelatin methacryloyl (GelMA), to obtain scaffolds with good mechanical properties for bone tissue engineering. The decellularized bone powder (DCB)-GelMA hybrid scaffolds had higher compressive strength and stiffness values when the DCB content was increased. In vitro evaluations revealed the biocompatibility of these scaffolds. The scaffolds could induce human bone marrow mesenchymal stem cells (hMSCs) to undergo osteogenic differentiation even in the absence of an induction medium. The efficiency of the scaffolds for bone regeneration applications was further evaluated using an in vivo cranial bone defect model in rats. Micro-CT images showed that the hybrid scaffolds with 20% DCB content had the best effect in promoting new bone regeneration. Thus, it was concluded that the DCB-GelMA hybrid scaffolds have high potential in bone tissue engineering applications.

Bioactive micropatterned platform to engineer myotube-like cells from stem cells.

Skeletal muscle has the capacity to repair and heal itself after injury. However, this self-healing ability is diminished in the event of severe injuries and myopathies. In such conditions, stem cell-based regenerative treatments can play an important part in post-injury restoration. We herein report the development of a bioactive (integrin-ß1antibody immobilized) gold micropatterned platform to promote human mesenchymal stem cell (hMSC) differentiation into myotube-like cells. hMSCs grown on bioactive micropattern differentiated into myotube-like cells within two weeks. Furthermore, the up-regulation of myogenic markers, multi-nucleated state with continuous actin cytoskeleton and the absence of proliferation marker confirmed the formation of myotube-like cells on bioactive micropattern. The prominent expression of elongated integrin-ß1(ITG-ß1) focal adhesions and the development of anisotropic stress fibers in those differentiated cells elucidated their importance in stem cell myogenesis. Together, these findings delineate the synergistic role of engineered cell anisotropy and ITG-ß1-mediated signaling in the development of myotube-like cells from hMSCs.

Highly substituted decoupled gelatin methacrylamide free of hydrolabile methacrylate impurities: An optimum choice for long-term stability and cytocompatibility.

Gelatin methacryloyl (GelMA; GM) contains impurities, including hydrolabile photosensitive methacrylate groups or soluble methacrylic acid (MA), which could be potentially detrimental to its in vitro and in vivo applications. To date, the influence of GM photocurable side chains on the cytotoxicity and ambient structural stability has remained to be investigated. Here, we successfully separated highly substituted decoupled gelatin methacrylamide (DGM) from GM via removing methacrylate impurities in order to evaluate its stability, cell viability, and cell toxicity, compared to GM, DGM plus soluble MA, and soluble MA. The photocurable methacrylate groups in GM were hydrolytically labile in neutral solutions, changing into soluble MA over time; on the other hand, the photocurable methacrylamide groups in DGM remained intact under the same conditions. Soluble MA was found to decrease cell viability in a dose dependent manner and caused severe cell toxicity at above 10 mg/mL. DGM plus MA started to impair cell viability at a 25 mg/mL concentration. DGM exhibited excellent cell viability and little cell toxicity across the treated concentrations (0.1-25 mg/mL). DGM without hydrolabile methacrylate and cytotoxic MA impurities could be a better choice for long term stability and good cell compatibility for bioapplications including bioprinting and cell encapsulation.

Inclusion of Cross-Linked Elastin in Gelatin/PEG Hydrogels Favourably Influences Fibroblast Phenotype.

The capacity of a biomaterial to innately modulate cell behavior while meeting the mechanical property requirements of the implant is a much sought-after goal within bioengineering. Here we covalently incorporate soluble elastin into a gelatin-poly (ethylene glycol) (PEG) hydrogel for three-dimensional (3D) cell encapsulation to achieve these properties. The inclusion of elastin into a previously optimized gelatin-PEG hydrogel was then evaluated for effects on entrapped fibroblasts, with the aim to assess the hydrogel as an extracellular matrix (ECM)-mimicking 3D microenvironment for cellular guidance. Soluble elastin was incorporated both physically and covalently into novel gelatin/elastin hybrid PEG hydrogels with the aim to harness the cellular interactivity and mechanical tunability of both elastin and gelatin. This design allowed us to assess the benefits of elastin-containing hydrogels in guiding fibroblast activity for evaluation as a potential dermal replacement. It was found that a gelatin-PEG hydrogel with covalently conjugated elastin, supported neonatal fibroblast viability, promoted their proliferation from 7.3% to 13.5% and guided their behavior. The expression of collagen alpha-1(COL1A1) and elastin in gelatin/elastin hybrid gels increased 16-fold and 6-fold compared to control sample at day 9, respectively. Moreover, cells can be loaded into the hydrogel precursor solution, deposited, and the matrix cross-linked without affecting the incorporated cells adversely, thus enabling a potential injectable system for dermal wound healing.

Photocurable Albumin Methacryloyl Hydrogels as a Versatile Platform for Tissue Engineering.

Photopolymerization of protein-derived polymers functionalized with methacryloyl groups has been increasingly used to fabricate three-dimensional tissue constructs for biomedical applications because photocurable protein-based polymers (e.g., gelatin and collagen methacryloyl) feature spatial-temporal controllability of engineering complex constructs as well as inherent biological properties. Herein, we report photocurable albumin-based hydrogels. First, photocurable bovine serum albumin methacryloyl (BSA-MA) with different degrees of substitution (DM) was successfully synthesized in a precise manner, without substantially altering BSA native secondary structure. Resultant photocurable BSA-MA hydrogels exhibited tunable physio-biochemical properties over the swelling, degradation, and mechanical properties. Moreover, photo-cross-linked BSA-MA hydrogels provided a permissible environment to support cell viability and functionality both in two- and three-dimensional culture systems. Photocurable BSA-MA hydrogels may be used as a versatile platform for various bioapplications including tissue engineering and 3D bioprinting.

Low Dose of Paclitaxel Combined with XAV939 Attenuates Metastasis, Angiogenesis and Growth in Breast Cancer by Suppressing Wnt Signaling.

Triple-negative breast cancer (TNBC) accounts for 15% of overall breast cancer. A lack of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2 receptor) makes TNBC more aggressive and metastatic. Wnt signaling is one of the important pathways in the cellular process; in TNBC it is aberrantly regulated, which leads to the progression and metastasis. In this study, we designed a therapeutic strategy using a combination of a low dose of paclitaxel and a Wnt signaling inhibitor (XAV939), and examined the effect of the paclitaxel-combined XAV939 treatment on diverse breast cancer lines including TNBC cell lines (MDA-MB-231, MDA-MB-468, and BT549) and ER+ve cell lines (MCF-7 and T-47D). The combination treatment of paclitaxel (20 nM) and XAV939 (10 µM) exerted a comparable therapeutic effect on MDA-MB-231, MDA-MB-468, BT549, MCF-7, and T-47D cell lines, relative to paclitaxel with a high dose (200 nM). The paclitaxel-combined XAV939 treatment induced apoptosis by suppressing Bcl-2 and by increasing the cleavage of caspases-3 and PARP. In addition, the in vivo results of the paclitaxel-combined XAV939 treatment in a mice model with the MDA-MB-231 xenograft further confirmed its therapeutic effect. Furthermore, the paclitaxel-combined XAV939 treatment reduced the expression of ß-catenin, a key molecule in the Wnt pathway, which led to suppression of the expression of epithelial-mesenchymal transition (EMT) markers and angiogenic proteins both at mRNA and protein levels. The expression level of E-cadherin was raised, which potentially indicates the inhibition of EMT. Importantly, the breast tumor induced by pristane was significantly reduced by the paclitaxel-combined XAV939 treatment. Overall, the paclitaxel-combined XAV939 regimen was found to induce apoptosis and to inhibit Wnt signaling, resulting in the suppression of EMT and angiogenesis. For the first time, we report that our combination approach using a low dose of paclitaxel and XAV939 could be conducive to treating TNBC and an external carcinogen-induced breast cancer.

5-hydroxymethylfurfural-embedded poly (vinyl alcohol)/sodium alginate hybrid hydrogels accelerate wound healing.

Treating large acute or chronic wounds remains a challenging task because of a lack of effective methods to accelerate wound healing. Though growth factor-based wound products are found to be effective, they are costly and are potentially associated with increased cancer mortality. Effective, safe, and affordable strategies to tackle large or chronic wounds need to further be designed and developed. Here, we designed a new antioxidant-embedded hydrogel system for speeding up the wound healing process. We prepared multifunctional poly (vinyl alcohol)/sodium alginate (PVA/SA) hydrogels with the incorporation of 5-hydroxymethylfurfural (5-HMF) and silver nanoparticles (Ag-NPs), and investigated their physiochemical and biological properties in vitro and in vivo. PVA, SA, 5-HMF, and Ag-NPs were employed for good mechanical properties, good biocompatibility, anti-inflammation, and anti-bacterial activity, respectively. 5-HMF is commonly found in many food products (e.g. honey, coffee, and black garlic) and is known as an antioxidant. In our in vitro study, 5-HMF was found to effectively facilitate the proliferation and migration of human skin fibroblasts (HSF), and collagen production. Besides, 5-HMF-embedded PVA/SA hybrid hydrogels supported controlled release and good cell compatibility, and more importantly accelerated wound healing in vivo through ameliorated inflammation, enhanced angiogenesis/vascularization, increased collagen production, and promoted re-epithelialization.

Epithelial-mesenchymal transition of cancer cells using bioengineered hybrid scaffold composed of hydrogel/3D-fibrous framework.

Cancer cells undergoing epithelial-mesenchymal transition (EMT) acquire stem cell-like phenotype associated with malignant behaviour, chemoresistance, and relapse. Current two-dimensional (2D) in-vitro culture models of tumorigenesis are inadequate to replicate the complexity of in-vivo microenvironment. Therefore, the generation of functional three-dimensional (3D) constructs is a fundamental prerequisite to form multi-cellular tumour spheroids for studying basic pathological mechanisms. In this study, we focused on two major points (i) designing and fabrication of 3D hybrid scaffolds comprising electrospun fibers with cancer cells embedded within hydrogels, and (ii) determining the potential roles of 3D hybrid scaffolds associated with EMT in cancer progression and metastasis. Our findings revealed that 3D hybrid scaffold enhances cell proliferation and induces cancer cells to undergo EMT, as demonstrated by significant up-regulation of EMT associated transcriptional factors including Snail1, Zeb1, and Twist2; and mesenchymal markers whereas epithelial marker, E-Cadherin was downregulated. Remarkably, this induction is independent of cancer cell-type as similar results were obtained for breast cancer cells, MDA-MB-231 and gastric cancer cells, MKN74. Moreover, the hybrid scaffolds enrich aggressive cancer cells with stem cell properties. We showed that our 3D scaffolds could trigger EMT of cancer cells which could provide a useful model for studying anticancer therapeutics against metastasis.