Development of injectable upconversion nanoparticle-conjugated doxorubicin theranostics electrospun nanostructure for targeted photochemotherapy in breast cancer.

Nanotheranostic-based photochemotherapies with targeted drug delivery have considerably surfaced in cancer therapy. In the presented work, polyethyleneimine-coated upconversion nanoparticles were engineered to conjugate covalently with doxorubicin. Upconversion nanoparticles (UCNP)-Doxorubicin (DOX)/synthesized epidermal growth factor receptor-targeting peptide blended with polymer composite was electrospun and formulated as the injectable dosage form. The size of the UCNP and the nanofiber diameter were assessed as 26.75 ± 1.54 and 162 ± 2.82 nm, respectively. The optimized ratio of dopants resulted in UCNP photoluminescence with maximum emission intensity at around 800 nm upon 980 nm excitation wavelength. The paramagnetic nature of UCNPs and amide conjugation with the drug was confirmed analytically. The loading capacity of UCNP for doxorubicin was determined to be 54.56%, while nanofibers exhibited 98.74% capacity to encapsulate UCNP-DOX. The release profile of UCNP-DOX from nanofiber formulation ranged from sustained to controlled, with relative enhancement in acidic conditions. The nanofiber demonstrated good mechanical strength, robust swelling, and degradation rate. Biocompatibility tests showed more than 90% cell viability on L929 and NIH/3T3 cell lines with UCNP-DOX@NF/pep nanoformulation. The IC50 values of 2.15 ± 0.54, 2.87 ± 0.67, and 3.42 ± 0.45 µg/mL on MDA-MB-231, 4T1, and MCF-7 cancer cell line, respectively, with a significant cellular uptake, has been reported. The UCNP protruded a ≈62.7°C temperature rise within 5 min of 980 nm laser irradiation and a power density of 0.5 W cm-2. The nanoformulation induced reactive oxygen species of 65.67% ± 3.21% and apoptosis by arresting the cell cycle sub-G1 phase. The evaluation conveys the effectiveness of the developed injectable theranostic delivery system in cancer therapy.

Natural bone inspired core-shell triple-layered gel/PCL/gel 3D printed scaffolds for bone tissue engineering.

Despite technological advancements in bone tissue engineering, it is still a challenge to fabricate a scaffold with high bioactivity as well as high mechanical strength that can promote osteogenesis as well as bear load. Here we developed a 3D printed gel-polymer multi-layered hybrid scaffold. The innermost layer is porous gel-based framework made of gelatin/carboxymethyl-chitin/nano-hydroxyapatite and is cryogenically 3D printed. Further, the second and middle layer of micro-engineered polycaprolactone (PCL) is infused in the gel with controlled penetration and tuneable coating thickness. The PCL surface is further coated with a third and final thin layer of gel matrix used for the first layer. This triple-layered structure demonstrates compression strength and modulus of 13.07 ± 1.15 MPa and 21.8 ± 0.82 MPa, respectively, post 8 weeks degradation which is >3000% and >700% than gel scaffold. It also shows degradation of 6.84 ± 0.70% (83% reduction than gel scaffold) after 12 weeks and swelling of 69.09 ± 6.83% (81% reduction) as compared to gel scaffolds. Further, nearly 300%, 250%, 50%, and 440% increase in cellular attachment, proliferation, protein generation, and mineralization, respectively are achieved as compared to only PCL scaffolds. Thus, these hybrid scaffolds offer high mechanical strength, slow degradation rate, high bioactivity, and high osteoconductivity. These multifunctional scaffolds have potential for reconstructing non-load-bearing bone defects like sinus lift, jaw cysts, and moderate load-bearing like reconstructing hard palate, orbital palate, and other craniomaxillofacial bone defects.

Theranostic magnetic nanoparticles enhance DNA damage and mitigate doxorubicin-induced cardio-toxicity for effective multi-modal tumor therapy.

The chemo-therapeutic efficacy of Doxorubicin (Dox), a potent anti-cancer drug used in the treatment of several solid tumors, is severely compromised by its cardio-toxicity. To overcome this shortcoming and exploit the utmost theranostic potential of nano-formulations, magnetic nanoparticles co-encapsulated with Dox and indocyanine green (ICG) in a liposomal carrier and tagged with cyclic RGD peptide were rationally designed and synthesized. These magneto-liposomes (T-LMD) showed αvß3-integrin receptor targeting and higher cyto-toxicity in several cancer cell lines (i.e. lung, breast, skin, brain and liver cancer) in combination with or without gamma radiation or magnetic hyperthermia therapy as compared to clinical liposomal nano-formulation of Dox (Lippod™). Mechanism of chemo-radio-sensitization was found to involve activation of JNK mediated pro-apoptotic signaling axis and delayed repair of DNA double strand breaks. Real time imaging of ICG labeled T-LMD suggested ~6-18 fold higher tumor accumulation of T-LMD as compared to off-target organs (kidney, liver, spleen, intestine, lungs and heart) and resulted in its higher combinatorial (chemo-radio-hyperthermia) tumor therapy efficacy as compared to Lippod™. Moreover, T-LMD showed insignificant toxicity to the heart tissue as suggested by serum levels of CK-MB, histo-pathological analysis, anti-oxidant enzyme activities (Catalase and GST) and markers of cardiac fibrosis, suggesting its potential for targeted multi-modal therapy of cancer.

"Viscotaxis"- directed migration of mesenchymal stem cells in response to loss modulus gradient.

Directed cell migration plays a crucial role in physiological and pathological conditions. One important mechanical cue, known to influence cell migration, is the gradient of substrate elastic modulus (E). However, the cellular microenvironment is viscoelastic and hence the elastic property alone is not sufficient to define its material characteristics. To bridge this gap, in this study, we investigated the influence of the gradient of viscous property of the substrate, as defined by loss modulus (G″) on cell migration. We cultured human mesenchymal stem cells (hMSCs) on a collagen-coated polyacrylamide gel with constant storage modulus (G') but with a gradient in the loss modulus (G″). We found hMSCs to migrate from high to low loss modulus. We have termed this form of directional cellular migration as "Viscotaxis". We hypothesize that the high loss modulus regime deforms more due to creep in the long timescale when subjected to cellular traction. Such differential deformation drives the observed Viscotaxis. To verify our hypothesis, we disrupted the actomyosin contractility with myosin inhibitor blebbistatin and ROCK inhibitor Y27632, and found the directional migration to disappear. Further, such time-dependent creep of the high loss material should lead to lower traction, shorter lifetime of the focal adhesions, and dynamic cell morphology, which was indeed found to be the case. Together, findings in this paper highlight the importance of considering the viscous modulus while preparing stiffness-based substrates for the field of tissue engineering. STATEMENT OF SIGNIFICANCE: While the effect of substrate elastic modulus has been investigated extensively in the context of cell biology, the role of substrate viscoelasticity is poorly understood. This omission is surprising as our body is not elastic, but viscoelastic. Hence, the role of viscoelasticity needs to be investigated at depth in various cellular contexts. One such important context is cell migration. Cell migration is important in morphogenesis, immune response, wound healing, and cancer, to name a few. While it is known that cells migrate when presented with a substrate with a rigidity gradient, cellular behavior in response to viscoelastic gradient has never been investigated. The findings of this paper not only reveal a completely novel cellular taxis or directed migration, it also improves our understanding of cell mechanics significantly.

Chemical Fingerprinting of Polymers Using Electron Energy-Loss Spectroscopy.

Electron energy-loss spectroscopy (EELS) is becoming an important tool in the characterization of polymeric materials. The sensitivity of EELS to changes in the chemical structure of polymeric materials dictates its applicability. In particular, it is important for compositional analysis to have reference spectra of pure components. Here, we report the spectra of the carbon K-edge of six polymers (polyethylene, polypropylene, polybutylene terephthalate, and polylactic acid) including copolymers (styrene acrylonitrile and acrylonitrile butadiene styrene), to be used as reference spectra for future EELS studies of polymers. We have successfully decomposed the carbon K-edge of each of the polymers and assigned the observed peaks to bonding transitions. The spectra have been acquired in standard experimental conditions, and electron beam damage has been taken into account during establishment of spectral-structural relationships. We found that the more commonly available low-energy resolution spectrometers are adequate to chemically fingerprint linear saturated hydrocarbons such as PE, PP, and PLA. We have thus moved a step closer toward creating an atlas of polymer EELS spectra, which can be subsequently used for chemical bond mapping of polymeric materials with nanoscale spatial resolution.

Viscoelastic substrate decouples cellular traction force from other related phenotypes.

Survival and maintenance of normal physiological functions depends on continuous interaction of cells with its microenvironment. Cells sense the mechanical properties of underlying substrate by applying force and modulate their behaviour in response to the resistance offered by the substrate. Most of the studies addressing cell-substrate mechanical interactions have been carried out using elastic substrates. Since tissues within our body are viscoelastic in nature, here we explore the effect of substrate's viscoelasticity on various properties of mesenchymal stem cells. Here, we used two sets of polyacrylamide substrates having similar storage modulus (G' = 1.1-1.6 kPa) but different loss modulus (G" = 45 Pa and 300 Pa). We report that human mesenchymal stem cells spread more but apply less force on the viscoelastic substrate (substrate with higher loss modulus). We further investigated the effect of substrate viscoelasticity on the expression of other contractility-associated proteins such as focal adhesion (FA) proteins (Vinculin, Paxillin, Talin), cytoskeletal proteins (actin, mysion, intermediate filaments, and microtubules) and mechano-sensor protein Yes-Associated Protein (YAP). Our results show that substrate viscoelasticity decouples cellular traction from other known traction related phenotypes.

Probing Kinetics and Mechanism of Formation of Mixed Metallic Nanoparticles in a Polymer Membrane by Galvanic Replacement between Two Immiscible Metals: Case Study of Nickel/Silver Nanoparticle Synthesis.

Galvanic replacement between metals has received notable research interest for the synthesis of heterometallic nanostructures. The growth pattern of the nanostructures depends on several factors such as extent of lattice mismatch, adhesive interaction between the metals, cohesive forces of the individual metals, etc. Due to the difficulties in probing ultrafast kinetics of the galvanic replacement reaction and particle growth in solution, real-time mechanistic investigations are often limited. As a result, the growth mechanism of one metal on the surface of another metal at the nanoscale is poorly understood so far. In the present work, we could successfully probe the galvanic replacement of silver ions with nickel nanoparticles, stabilized in a polymer membrane, using two complementary methods, namely, small-angle X-ray scattering (SAXS) and radiolabeling, and the results are supported by density functional theory (DFT) computations. The silver-nickel system has been chosen for the present investigation because of the high degree of bulk immiscibility caused by the large lattice mismatch (15.9%) and the weak adhesive interaction, which makes it a perfect model system for immiscible metal pairs. Membrane, as a host medium, plays a crucial role in retarding the kinetics of atomic and particle rearrangements (nucleation and growth) due to slower mobility of the atoms (monomers) and particles within the polymer network. This allowed us to examine the real-time concentration of silver monomers during galvanic replacement of silver ions with nickel nanoparticles and evolution of Ni/Ag nanoparticles. From combined experiment and DFT computations, it has been demonstrated, for the first time to the best of our knowledge, that the majority of silver atoms, which are produced on the nickel nanoparticle surface by galvanic reactions, do not form traditional core-shell nanostructures with nickel and undergo a self-governing sequential nucleation and growth of silver nanoparticles via formation of intermediate prenucleation silver clusters, leading to the formation of mixed metallic nanoparticles in the membrane. The surface of NiNPs has a heterogeneous effect on the silver nucleation pathway, which is evident from the reduced critical free energy barrier of nucleation (ΔGcrit). The present work establishes an original mechanistic pathway based on a sequential nucleation model for formation of mixed metallic nanoparticles by the galvanic replacement route, which opens up future possibilities for size-controlled synthesis in mixed systems.

Ayurvedic processing of α-HgS gives novel physicochemistry and distinct toxicokinetics in zebrafish.

Rasasindura (RS) is an Ayurvedic medicine, which contains ∼99% α-HgS. It is used as a rejuvenating agent and commonly used to treat diseases such as syphilis, insomnia, high fever, and nervous disorders. Cinnabar ore (α-HgS) is a well-known mineral, which is readily available. Despite it, Ayurvedic practitioners adopted an involved and tedious procedure for the preparation of RS. In this study, three samples, one was Ayurvedic (RS), the second one was the commercial (HGS), and the third one was cinnabar ore (CN), were physiochemically examined. Zebrafish model was employed for toxicity study with an oral dose of 100 mg/kg/day for the three samples for 10 days. We found that RS conferred novel physicochemical properties, which were not seen in HGS and CN. Significantly, the average crystallite size of RS was lowest (26 nm) as compared to HGS (31 nm) and CN (34 nm), and the rate of increase of crystallite size with temperature was lowest in RS. RS did not show any significant behavioral toxicity in zebrafish, which was seen with the HGS-and CN-treated zebrafish. HGS-and CN-treated zebrafish showed a significantly high (∗∗∗p < 0.001) decrease (77 ± 7.6% and 51 ± 6.5%, respectively) of glutathione (GSH) levels in the brain, however, for RS-treated zebrafish, the change of GSH was insignificant (26 ± 2.5%, p > 0.05). Interestingly, HGS significantly altered the γ-aminobutyric acid (GABA) in brain tissue. Therefore, among all three samples, RS exhibited the lowest toxicity, which can be credited to the distinct toxicokinetics by these samples.

Surface-Functionalized Poly(Ether Sulfone) Composite Hollow Fiber Membranes with Improved Biocompatibility and Uremic Toxins Clearance for Bioartificial Kidney Application.

Bioartificial kidney (BAK) is attracting the focus of the research community. In this study, the efficacy of surface-functionalized poly(ether sulfone)-TPGS-graphene oxide composite hollow fiber membranes as a promising material for the single extracorporeal unit for BAK application was evaluated. The cytotoxicity was examined using human primary renal proximal tubular epithelial cells (hPTCs), and the removal of uremic toxins (urea, creatinine, phosphate, and lysozyme) from the toxin-spiked goat blood was measured. The surface-functionalized polymer composite membranes acted as a biocompatible material for attachment and proliferation of hPTCs, which was confirmed by microscopy studies, proliferation, and activity assays. The functional activity of these renal cells over this biocompatible membrane was also maintained. Remarkably, the functionalized composite membranes showed removal of urea (46.4 ± 3.5%), creatinine (52.2 ± 3.9%), phosphate (35.5 ± 2.7%), and lysozyme (11.2 ± 0.8%) from the toxin-spiked goat blood. Therefore, these obtained results showed that the surface-functionalized poly(ether sulfone)-TPGS-graphene oxide composite hollow fiber membranes are suitable for BAK application.

Accentuated osseointegration in osteogenic nanofibrous coated titanium implants.

Anchoring of endosseous implant through osseointegration continues to be an important clinical need. Here, we describe the development of superior endosseous implant demonstrating enhance osseointegration, achieved through surface modification via coating of osteogenic nanofibres. The randomized bio-composite osteogenic nanofibres incorporating polycaprolactone, gelatin, hydroxyapatite, dexamethasone, beta-glycerophosphate and ascorbic acid were electrospun on titanium implants mimicking bone extracellular matrix and subsequently induced osteogenesis by targeting undifferentiated mesenchymal stem cells present in the peri-implant niche to regenerate osseous tissue. In proof-of-concept experiment on rabbit study models (n = 6), micro-computed tomography (Micro-CT), histomorphometric analysis and biomechanical testing in relation to our novel osteogenic nanofibrous coated implants showed improved results when compared to uncoated controls. Further, no pathological changes were detected during gross examination and necropsy on peri-implant osseous tissues regenerated in response to such coated implants. The findings of the present study confirm that osteogenic nanofibrous coating significantly increases the magnitude of osteogenesis in the peri-implant zone and favours the dynamics of osseointegration.

Polyethersulfone-carbon nanotubes composite hollow fiber membranes with improved biocompatibility for bioartificial liver.

Carbon nanotubes (CNTs) blended hollow fiber membranes (HFMs) are a promising new material in the area of biomedical engineering because they simultaneously provide tunable hydrophilicity along with selective permeability. In the present study, composite polyethersulfone (P) HFMs were fabricated using d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS or T) as compatibilizer, and carboxylated multiwalled CNTs (MWCNTs or C) as filler. The amount of MWCNTs was optimized for the improved hemocompatibility, cell viability, and cellular functionality. An optimum was found with the composte HFMs (PTC-2), where MWCNTs were used at concentration of 0.030 wt.%, as it exhibited improved compatibility with human blood. Further, these PTC-2 HFMs showed enhanced liver (HepG2) cells growth with the enhanced cell functional activities, mainly albumin secretion and glucose consumption. These developed composite membrane can act as a membrane material for liver cell bioreactor and bioartificial liver development because of their 3D scaffold like characteristic which enables cell growth, and selective permeability which helps in immunoisolation.

Multiscale Porosity in Compressible Cryogenically 3D Printed Gels for Bone Tissue Engineering.

Three-dimensional (3D) printing technology has seen several refinements when introduced in the field of medical devices and regenerative medicines. However, it is still a challenge to 3D print gels for building complex constructs as per the desired shape and size. Here, we present a novel method to 3D print gelatin/carboxymethylchitin/hydroxyapatite composite gel constructs of a complex shape. The objective of this study is to fabricate a bioactive gel scaffold with a controlled hierarchical structure. The hierarchy ranges from 3D outer shape to macroporosity to microporosity and rough surface. The fabrication process developed here uses 3D printing in a local cryogenic atmosphere, followed by lyophilization and cross-linking. The gel instantly freezes after extrusion on the cold plate. The cooling action is not limited to the build plate, but the entire gel scaffold is cooled during the 3D printing process. This enables the construction of a stable self-sustaining large-sized 3D complex geometry. Further, lyophilization introduces bulk microporosity into the scaffolds. The outer shape and macroporosity were controlled with the 3D printer, whereas the microporous structure and desirable rough surface morphology were obtained through lyophilization. With cryogenic 3D printing, up to 90% microporosity could be incorporated into the scaffolds. The microporosity and pore size distribution were controlled by changing the cross-linker and total polymer concentration, which resulted in six times increase in surface open pores of size <20 µm on increasing the cross-linker concentration from 25 to 100 mg/mL. The introduction of bulk microporosity was shown to increase swelling by 1.8 times along with a significant increase in human umbilical cord mesenchymal stem cells and Saos-2 cell attachment (2×), proliferation (2.4×), Saos-2 cell alkaline phosphatase level (2×), and mineralization (3×). The scaffolds are spongy in nature in a wet state, thus making them potential implants for bone cavities with a small opening. The application of these cryogenically 3D printed compressible gel scaffolds with multiscale porosity extends to a small- as well as a large-sized open/partially open patient-specific bone defect.

Modelling and optimization of NaOH-etched 3-D printed PCL for enhanced cellular attachment and growth with minimal loss of mechanical strength.

Despite having gained success in achieving intricate geometries for bone-graft fabrication, 3D printing technology still lacks good implant-tissue bonding. This can be addressed with alkaline surface post-treatment of 3D printed grafts, which improves the surface morphology and cellular response (attachment and proliferation), as shown in this study of polycaprolactone (PCL). The parameters for process optimization were NaOH-concentration, reaction temperature, and treatment time. Along with the hydrolysis reaction, its morphological implications at micro-level was also studied here for the first time. The modified surface was characterized by measuring surface porosity, surface roughness, and cellular response. A kinetic model was developed to correlate surface porosity with concentration, temperature and time. The concept of treatment intensity is introduced, which is a lumped parameter consisting of the product of the three governing parameters, which shows a concentration-temperature-time equivalency. With the increase in treatment intensity, surface porosity increased to ~60%, the surface roughness (RMS value) increased to ~700 nm, and cellular response improved till surface porosity reaches ~35%. This study establishes the importance of NaOH-PCL interaction and proposes that the surface reaction mechanism studied here can be exploited to enhance the in-vivo performance of bone grafts.

A biodegradable fluorescent nanohybrid for photo-driven tumor diagnosis and tumor growth inhibition.

Specific targeting and phototriggered therapy in mouse model have recently emerged as the starting point of cancer theragnosis. Herein, we report a bioresponsive and degradable nanohybrid, a liposomal nanohybrid decorated with red emissive carbon dots, for localized tumor imaging and light-mediated tumor growth inhibition. Unsaturated carbon dots (C-dots) anchored to liposomes convert near-infrared (NIR) light into heat and also produce reactive oxygen species (ROS), demonstrating the capability of phototriggered cancer cell death and tumor regression. The photothermal and oxidative damage of breast tumor by the nonmetallic nanohybrid has also been demonstrated. Designed nanoparticles show excellent aqueous dispersibility, biocompatibility, light irradiated enhanced cellular uptake, release of reactive oxygen species, prolonged and specific tumor binding ability and good photothermal response (62 °C in 5 minutes). Safe and localized irradiation of 808 nm light demonstrates significant tumor growth inhibition and bioresponsive degradation of the fluorescent nanohybrid without affecting the surrounding healthy tissues.

Matrix-entrapped cellular secretome rescues diabetes-induced EPC dysfunction and accelerates wound healing in diabetic mice.

Cellular secretory products have infinite potential, which is only recently explored for research and therapeutic applications. The present study elaborated on the formation of a unique matrix-entrapped cellular secretome (MCS), a hydrogel-like secretome produced by bone marrow-derived mononuclear cells when cultured on a three-dimensional electrospun nanofiber matrix under specific conditions. These culture conditions support the growth of a mixed population predominantly comprising of endothelial precursor cells (EPCs), along with mesenchymal stromal cells and pericytes. Interestingly, such secretome is not formed in a pure culture of EPCs on the similarly formulated matrix, suggesting that a heterotypic cell-cell interaction is essential for the formation of MCS. In addition, the specific composition of the matrix was found to be a critical necessity for the formation of MCS. Furthermore, the application of the MCS as a substrate promotes the growth of EPCs in culture. It also rescues the diabetes-induced EPC dysfunction as assessed based on the parameters, such as viability, proliferation, colony formation, cellular adhesion, chemotactic migration, and tubule formation. MCS augments the levels of eNOS-specific mRNA (Nos3) and also promotes the restoration of the SDF1/CXCR4 axis in diabetic EPCs. Notably, a topical application of MCS on diabetic wounds leads to an accelerated wound closure. Thus, the current data showed that MCS forms an excellent cell-free biomaterial in the treatment of diabetic wounds and non-healing ulcers.

Gloriosa superba Mediated Synthesis of Platinum and Palladium Nanoparticles for Induction of Apoptosis in Breast Cancer.

Green chemistry approaches for designing therapeutically significant nanomedicine have gained considerable attention in the past decade. Herein, we report for the first time on anticancer potential of phytogenic platinum nanoparticles (PtNPs) and palladium nanoparticles (PdNPs) using a medicinal plant Gloriosa superba tuber extract (GSTE). The synthesis of the nanoparticles was completed within 5 hours at 100°C which was confirmed by development of dark brown and black colour for PtNPs and PdNPs, respectively, along with enhancement of the peak intensity in the UV-visible spectra. High-resolution transmission electron microscopy (HRTEM) showed that the monodispersed spherical nanoparticles were within a size range below 10 nm. Energy dispersive spectra (EDS) confirmed the elemental composition, while dynamic light scattering (DLS) helped to evaluate the hydrodynamic size of the particles. Anticancer activity against MCF-7 (human breast adenocarcinoma) cell lines was evaluated using MTT assay, flow cytometry, and confocal microscopy. PtNPs and PdNPs showed 49.65 ± 1.99% and 36.26 ± 0.91% of anticancer activity. Induction of apoptosis was most predominant in the underlying mechanism which was rationalized by externalization of phosphatidyl serine and membrane blebbing. These findings support the efficiency of phytogenic fabrication of nanoscale platinum and palladium drugs for management and therapy against breast cancer.

Hydrophilic ZIF-8 decorated GO nanosheets improve biocompatibility and separation performance of polyethersulfone hollow fiber membranes: A potential membrane material for bioartificial liver application.

The hydrophobic nature of zeolitic imidazole framework-8 (ZIF-8) nanoparticles restricts their use as additives in hollow fiber membranes (HFMs) for biomedical applications. In this study, hydrophilic ZIF-8 decorated graphene oxide nanosheets (ZGs) were synthesized and used as additives (0-1 wt%) in polyethersulfone (P) HFMs with the aim of improving the biocompatibility and separation performance so as to make the ZGP HFMs suitable for bioartificial liver (BAL) application. Elemental mapping and Fourier transform infrared studies confirmed the efficacious incorporation of ZG nanohybrids in the ZGP HFMs, which resulted in their improved hydrophilicity. The remarkably improved biocompatibility was experimentally demonstrated for the ZGP HFMs, which also were antioxidative and hemocompatible. There was a significantly high attachment and proliferation of HepG2 cells on these HFMs, and they showed remarkably high urea synthesis and albumin secretion. Further, the ZGP HFMs showed high ultrafiltration coefficient (392.2 ±â€¯26.5 mL/h/m2/mm Hg), high flux recovery ratio (84.3%), low flux reduction (15.7%), and desirable molecular weight cutoff (125-135 kDa). Thus, these results experimentally demonstrated that the hydrophilic ZG nanohybrids improve the desirable properties of ZGP HFMs making them a potential biocompatible material for biomedical applications including BAL application.

Development of hybrid scaffold with biomimetic 3D architecture for bone regeneration.

In the present study, a biomimetic three-dimensional hybrid scaffold has been designed considering the bone natural architecture with favorable interconnected porous structure, nano-microscale features and mechanical strength. The chief components of the hybrid scaffold are core-sheath nanofibers and hydrogel, suitably arranged to create a bone like microenvironment. Specifically, the core-sheath nanofibers were coiled tightly into a ring to mimic the osteon, and reinforced in a hydrogel matrix. Morphological analysis using SEM and 4D-X-ray microscopy revealed that the hybrid scaffold consists of coiled rings of nanofibers in highly porous hydrogel matrix showing structural similarity to osteons. The reinforcement of electrospun nanofibers in hydrogel influenced the mechanical properties of scaffold. The potential application of the biomimetic hybrid scaffold, and the role of its specific architecture, was subsequently investigated in vitro using a human osteosarcoma fibroblast cell line. Furthermore, DNA quantification, alkaline-phosphatase and alizarin assay validated the potential of fabricated scaffold for bone tissue-regeneration.

Functionally coated polyethersulfone hollow fiber membranes: A substrate for enhanced HepG2/C3A functions.

Hollow fiber membrane (HFM) based liver assist systems are a life-saving bridge for patients until a donor organ is available for transplantation or until liver regeneration. However, liver cell attachment and functional maintenance on HFM surface is a major challenge in bio-artificial liver (BAL) support systems. In the present study, novel glutaraldehyde (GTA)-crosslinked gelatin (gel)-coated polyethersulfone (X-gel-PT) HFMs were manufactured using triple orifice spinneret by the dry-wet spinning method. HFMs were characterized for morphology, outer surface roughness, hydrophilicity, tensile strength, thermal stability, BET surface area and pore volume measurements, permeability and rejection. Fourier transform infrared spectroscopy, and transmission electron microscopy confirmed the GTA-crosslinked gel-coating in the X-gel-PT HFMs, which provided the desirable extracellular matrix-like environment to the HepG2/C3A cells. The results of in-vitro hemocompatibility tests showed the better suitability of the developed HFMs for the blood-contact application. X-gel-PT HFMs showed significantly better cellular attachment and proliferation of HepG2/C3A cells on day 3 and 6, as shown by scanning electron and confocal microscopy. Significantly high urea synthesis and albumin secretion seen indicated the improved functional and metabolic activity of HepG2/C3A cells. Thus, the developed X-gel-PT HFMs is a suitable substrate for the hepatocyte culture, mass culture, and development of BAL support system.

Three-dimensional multiscale fiber matrices: development and characterization for increased HepG2 functional maintenance for bio-artificial liver application.

The development of a cell-growth substrate that provides a nature-like microenvironment, promotes cell adhesion, and maintains the cells' functional activities is a research focus in the field of tissue engineering. In the present study, three-dimensional micro-nano multiscale fiber-based substrates were developed by depositing biocompatible polycaprolactone (PCL)/PCL-Chitosan (C)/PCL-C-Gelatin (G) electrospun nanofibers (NFs) on the outer surface of hollow fiber membranes (HFMs) in one step. A comparison study with regard to physico-chemical characterization, hemocompatibility, cytotoxicity, and cellular functionality was performed with the developed matrices. The PCL-C-G NFs-deposited HFMs-based matrix showed superior hemocompatibility for blood-contact applications. The cytotoxicity of these matrices was found to be minimal. HepG2 cells exhibited an exceptionally robust adherence and proliferated growth on the matrix with the formation of characteristic multi-cellular spheroids. Furthermore, cell functional activities such as albumin secretion, urea synthesis, and cytochrome P450 specific activity were measured for the developed matrices. The developed three-dimensional multiscale fibers-based matrix can be a potential membrane for bioreactor and bio-artificial liver applications.