Doxorubicin Conjugated γ-Globulin Functionalised Gold Nanoparticles: A pH-Responsive Bioinspired Nanoconjugate Approach for Advanced Chemotherapeutics.

Developing successful nanomedicine hinges on regulating nanoparticle surface interactions within biological systems, particularly in intravenous nanotherapeutics. We harnessed the surface interactions of gold nanoparticles (AuNPs) with serum proteins, incorporating a γ-globulin (γG) hard surface corona and chemically conjugating Doxorubicin to create an innovative hybrid anticancer nanobioconjugate, Dox-γG-AuNPs. γG (with an isoelectric point of ~7.2) enhances cellular uptake and exhibits pH-sensitive behaviour, favouring targeted cancer cell drug delivery. In cell line studies, Dox-γG-AuNPs demonstrated a 10-fold higher cytotoxic potency compared to equivalent doxorubicin concentrations, with drug release favoured at pH 5.5 due to the γ-globulin corona's inherent pH sensitivity. This bioinspired approach presents a novel strategy for designing hybrid anticancer therapeutics. Our study also explored the intricacies of the p53-mediated ROS pathway's role in regulating cell fate, including apoptosis and necrosis, in response to these treatments. The pathway's delicate balance of ROS emerged as a critical determinant, warranting further investigation to elucidate its mechanisms and implications. Overall, leveraging the robust γ-globulin protein corona on AuNPs enhances biostability in harsh serum conditions, augments anticancer potential within pH-sensitive environments, and opens promising avenues for bioinspired drug delivery and the design of novel anticancer hybrids with precise targeting capabilities.

Biocidal polymer derived near white light-emitting polymeric carbon particles for antibacterial and bioimaging applications.

A growing antimicrobial crisis has increased demand for antimicrobial materials. It has become increasingly popular to convert polymeric macromolecules into polymeric carbon particles (PCP) in order to achieve highly biocompatible materials with unique properties as a result of the ability to synthesize nanomaterials of the right size and add value to existing stable polymers. This work presents the tuning of PCP for antibacterial application by combining a biocidal polymer with one-pot solvothermal synthesis. PCP displayed broad-spectrum antibacterial activity via various mechanisms, including inhibition of bacterial cell walls, ROS generation, and antibiotic resistance. Furthermore, these biocidal PCP were observed to show excitation-independent near-white light emission which on the other hand is generally possible due to mixed sizes, doping, and surface effects. As opposed to the parent biocidal polymer, PCP added ROS-mediated bactericidal activity, increased cytocompatibility, and nanofibers with anti-adhesive effects and potential of imaging bacterial cells.

Post-Implantation Stiffening by a Bioinspired, Double-Network, Self-Healing Hydrogel Facilitates Minimally Invasive Cell Delivery for Cartilage Regeneration.

Injectable hydrogels have demonstrated advantages in cartilage repair by enabling the delivery of cells through a minimally invasive approach. However, several injectable hydrogels suffer from rapid degradation and low mechanical strength. Moreover, higher mechanical stiffness in hydrogels can have a detrimental effect on post-implantation cell viability. To address these challenges, we developed an in situ forming bioinspired double network hydrogel (BDNH) that exhibits temperature-dependent stiffening after implantation. The BDNH mimics the microarchitecture of aggrecan, with hyaluronic acid-conjugated poly(N-isopropylacrylamide) providing rigidity and Schiff base crosslinked polymers serving as the ductile counterpart. BDNHs exhibited self-healing property and enhanced stiffness at physiological temperature. Excellent cell viability, long time cell proliferation, and cartilage specific matrix production were observed in the chondrocytes cultured in the BDNH hydrogel. Evidence of cartilage regeneration in a rabbit cartilage defect model using chondrocyte-laden BDNH has suggested it to be a potential candidate for cartilage tissue engineering.

Injectable Bone Cement Reinforced with Gold Nanodots Decorated rGO-Hydroxyapatite Nanocomposites, Augment Bone Regeneration.

Interest in the development of new generation injectable bone cements having appropriate mechanical properties, biodegradability, and bioactivity has been rekindled with the advent of nanoscience. Injectable bone cements made with calcium sulfate (CS) are of significant interest, owing to its compatibility and optimal self-setting property. Its rapid resorption rate, lack of bioactivity, and poor mechanical strength serve as a deterrent for its wide application. Herein, a significantly improved CS-based injectable bone cement (modified calcium sulfate termed as CSmod ), reinforced with various concentrations (0-15%) of a conductive nanocomposite containing gold nanodots and nanohydroxyapatite decorated reduced graphene oxide (rGO) sheets (AuHp@rGO), and functionalized with vancomycin, is presented. The piezo-responsive cement exhibits favorable injectability and setting times, along with improved mechanical properties. The antimicrobial, osteoinductive, and osteoconductive properties of the CSmod cement are confirmed using appropriate in vitro studies. There is an upregulation of the paracrine signaling mediated crosstalk between mesenchymal stem cells and human umbilical vein endothelial cells seeded on these cements. The ability of CSmod to induce endothelial cell recruitment and augment bone regeneration is evidenced in relevant rat models. The results imply that the multipronged activity exhibited by the novel-CSmod cement would be beneficial for bone repair.

A drug-free strategy to combat bacterial infections with magnetic nanoparticles biosynthesized in bacterial pathogens.

The extensive and indiscriminate use of antibiotics in the ongoing COVID-19 pandemic might significantly contribute to the growing number of multiple drug resistant (MDR) bacteria. With the dwindling pipeline of new and effective antibiotics, we might soon end up in a post-antibiotic era, in which even common bacterial infections would be a challenge to control. To prevent this, an antibiotic-free strategy would be highly desirable. Magnetic nanoparticle (MNP)-mediated hyperthermia-induced antimicrobial therapy is an attractive option as it is considered safe for human use. Given that iron and zinc are critical for bacterial virulence, we evaluated the response of multiple pathogenic bacteria to these elements. Treatment with 1 mM iron and zinc precursors resulted in the intracellular biosynthesis of MNPs in multiple Gram-positive and Gram-negative disease-causing bacteria. The superparamagnetic nanoparticles in the treated bacteria/biofilms, generated heat upon exposure to an alternating magnetic field (AMF), which resulted in an increase in the temperature (5-6 °C) of the milieu with a subsequent decrease in bacterial viability. Furthermore, we observed for the first time that virulent bacteria derived from infected samples harbour MNPs, suggesting that the bacteria had biosynthesised the MNPs using the metal ions acquired from the host. AMF treatment of the bacterial isolates from the infected specimens resulted in a strong reduction in viability (3-4 logs) as compared to vancomycin/ciprofloxacin treatment. The therapeutic efficacy of the MNPs to induce bacterial death with AMF alone was confirmed ex vivo using infected tissues. Our proposed antibiotic-free approach for killing bacteria using intracellular MNPs is likely to evolve as a promising strategy to combat a wide range of bacterial infections.

Mechanical Integrity in a Dynamic Interpenetrating Hydrogel Network of Supramolecular Peptide-Polysaccharide Supports Enhanced Chondrogenesis.

Tissue engineering demands intelligently designed scaffolds that encompass the properties of the target tissues in terms of mechanical and bioactive properties. An ideal scaffold for engineering a cartilage tissue should provide the chondrocytes with a favorable 3D microarchitecture apart from possessing optimal mechanical characteristics such as compressibility, energy dissipation, strain stiffening, etc. Herein, we used a unique design approach to develop a hydrogel having a dynamic interpenetrating network to serve as a framework to support chondrocyte growth and differentiation. An amyloid-inspired peptide amphiphile (1) was self-assembled to furnish kinetically controlled nanofibers and incorporated in a dynamic covalently cross-linked polysaccharide network of carboxymethyl cellulose dialdehyde (CMC-D) and carboxymethyl chitosan (CMCh) using Schiff base chemistry. The dynamic noncovalent interaction played a pivotal role in providing the desired modulation in the structure and mechanical properties of the double-network hydrogels that are imperative for cartilage scaffold design. The adaptable nature supported shear-induced extrusion of the hydrogel and facilitated various cellular functions while maintaining its integrity. The potential of the as-developed hydrogels to support in vitro chondrogenesis was explored using human chondrocytes. Evidence of improved cell growth and cartilage-specific ECM production confirmed the potential of the hydrogel to support cartilage tissue engineering while reaffirming the significance of mimicking the biophysical microenvironment to induce optimal tissue regeneration.

An injectable hydrogel having proteoglycan-like hierarchical structure supports chondrocytes delivery and chondrogenesis.

The ECM of cartilage is composed of proteoglycans (PG) that contain glycosaminoglycan (GAG), aggrecan, hyaluronic acid (HA) and other molecular components which play an important role in regulating chondrocyte functions via cell-matrix interactions, integrin-mediated signalling etc. Implantation of chondrocytes encapsulated in scaffolds that mimic the micro-architecture of proteoglycan, is expected to enhance cartilage repair. With an aim to create a hydrogel having macromolecular structure that resembles the cartilage-specific ECM, we constructed a hierarchal structure that mimic the PG. The bottle brush structure of the aggrecan was obtained using chondroitin sulphate and carboxymethyl cellulose which served as GAG and core protein mimic respectively. A proteoglycan-like structure was obtained by cross-linking it with modified chitosan that served as a HA substitute. The physico-chemical characteristics of the above cross-linked injectable hydrogel supported long term human articular chondrocyte subsistence and excellent post-injection viability. The chondrocytes encapsulated in the PMH expressed significant levels of articular cartilage specific markers like collagen II, aggrecan, GAGs etc., indicating the ability of the hydrogel to support chondrocyte differentiation. The biocompatibility and biodegradability of the hydrogels was confirmed using suitable in vivo studies. The results revealed that the PG-mimetic hydrogel could serve as a promising scaffold for chondrocyte implantation.

Tunable, conductive, self-healing, adhesive and injectable hydrogels for bioelectronics and tissue regeneration applications.

Conductive hydrogels are attracting considerable interest in view of their potential in a wide range of applications that include healthcare and electronics. Such hydrogels are generally incorporated with conductive materials/polymers. Herein, we present a series of conductive hydrogels (Ch-CMC-PDA), prepared with no additional conductive material. The hydrogels were synthesized using a combination of chitosan, cellulose (CMC) and dopamine (DA). The conductivity (0.01-3.4 × 10-3 S cm-1) in these gels is attributed to ionic conductivity. Very few conductive hydrogels are endowed with additional properties like injectability, adhesiveness and self-healing, which would help to widen their scope for applications. While the dynamic Schiff base coupling in our hydrogels facilitated self-healing and injectable properties, polydopamine imparted tissue adhesiveness. The porosity, rheological, mechanical and conductive properties of the hydrogels are regulated by the CMC-dialdehyde-polydopamine (CMC-D-PDA) content. The hydrogel was evaluated in various bioelectronics applications like ECG monitoring and triboelectric nanogenerators (TENG). The ability of the hydrogel to support cell growth and serve as a template for tissue regeneration was confirmed using in vitro and in vivo studies. In summary, the integration of such remarkable features in the ionic-conductive hydrogel would enable its usage in bioelectronics and biomedical applications.

Actin-binding carbon dots selectively target glioblastoma cells while sparing normal cells.

Curcumin, a pleiotropic signalling molecule from Curcuma longa, is reported to be effective against multiple cancers. Despite its promising effect, curcumin had failed in clinical trials due to its low aqueous solubility, stability and poor bioavailability. While several approaches are being attempted to overcome the limitations, the improved solubility observed with curcumin-derived carbon dots appeared to be a strategy worth exploring. To assess if the carbon dots possess bio-activity similar to curcumin, we synthesized carbon dots (CurCD) from curcumin and ethylenediamine. Unlike curcumin, the as-synthesized curcumin carbon dots exhibited excellent solubility, excitation-dependent emission and photostability. The anti-cancer activity evaluated with glioblastoma cells using the well-established in vitro models indicated its comparable/enhanced activity over curcumin. Besides, the selective affinity of CurCD to the actin filament, indicated it's prospective to serve as a marker of actin filaments. In addition, the non-toxic effects observed in normal cells and fish embryos indicated CurCD was more biocompatible than curcumin. While this work reveals the superior properties of CurCD over curcumin, it provides a new approach to explore other plant derived molecules with similar limitations like curcumin.

A bioinspired, ice-templated multifunctional 3D cryogel composite crosslinked through in situ reduction of GO displayed improved mechanical, osteogenic and antimicrobial properties.

3D biopolymeric scaffolds often lack the biochemical cues and mechanical strength to encourage bone tissue regeneration. Chemical crosslinkers have been extensively used to impart strength, but have been found to be toxic at the site of implantation and possess a lacuna in physical strength. We attempted to address this by engineering a self-crosslinked polymer through the in-situ reduction of Graphene oxide (GO) in a gelatin cryogel (Gel-RGO) using ice as a template to create pores. Superior osteoinductive and antimicrobial properties were further endowed on the cryogel by incorporating silver nanoparticles decorated nanohydroxyapatite in the Gel-RGOAg@Hap(2%) cryogel. The optimized biocompatible cryogel favoured bone cell adhesion and its proliferation. The osteoconductive and osteoinductive potential of the cryogel was confirmed through biomineralization and differentiation of bone cells. In addition, these cryogels showed prolonged antimicrobial activity against S. aureus. This investigation exhibits the achievability/prospect of building up an ideal gelatin platform without the utilization of an outside crosslinking agent via manipulating the conditions of gelation. The superior crosslinking achieved between gelatin and GO, in addition to its ability to support bone formation and prevent infection make this cryogel an attractive candidate for bone tissue engineering applications.

Synthesis and Evaluation of a Zinc Eluting rGO/Hydroxyapatite Nanocomposite Optimized for Bone Augmentation.

Repair of critical size bone defects is a clinical challenge that usually necessitates the use of bone substitutes. For successful bone repair, the substitute should possess osteoconductive, osteoinductive, and vascularization potential, with the ability to control post-implantation infection serving as an additional advantage. With an aim to develop one such substitute, we optimized a zinc-doped hydroxyapatite (HapZ) nanocomposite decorated on reduced graphene oxide (rGO), termed as G3HapZ, and demonstrated its potential to augment the bone repair. The biocompatible composite displayed its osteoconductive potential in biomineralization studies, and its osteoinductive property was confirmed by its ability to induce mesenchymal stem cell (MSC) differentiation to osteogenic lineage assessed by in vitro mineralization (Alizarin red staining) and expression of osteogenic markers including runt-related transcription factor 2 (RUNX-2), alkaline phosphatase (ALP), type 1 collagen (COL1), bone morphogenic protein-2 (BMP-2), osteocalcin (OCN), and osteopontin (OPN). While the potential of G3HapZ to support vascularization was displayed by its ability to induce endothelial cell migration, attachment, and proliferation, its antimicrobial activity was confirmed using S. aureus. Biocompatibility of G3HapZ was demonstrated by its ability to induce bone regeneration and neovascularization in vivo. These results suggest that G3HapZ nanocomposites can be exploited for a range of strategies in developing orthopedic bone grafts to accelerate bone regeneration.

Nanotechnology for COVID-19: Therapeutics and Vaccine Research.

The current global health threat by the novel coronavirus disease 2019 (COVID-19) requires an urgent deployment of advanced therapeutic options available. The role of nanotechnology is highly relevant to counter this "virus" nano enemy. Nano intervention is discussed in terms of designing effective nanocarriers to counter the conventional limitations of antiviral and biological therapeutics. This strategy directs the safe and effective delivery of available therapeutic options using engineered nanocarriers, blocking the initial interactions of viral spike glycoprotein with host cell surface receptors, and disruption of virion construction. Controlling and eliminating the spread and reoccurrence of this pandemic demands a safe and effective vaccine strategy. Nanocarriers have potential to design risk-free and effective immunization strategies for severe acute respiratory syndrome coronavirus 2 vaccine candidates such as protein constructs and nucleic acids. We discuss recent as well as ongoing nanotechnology-based therapeutic and prophylactic strategies to fight against this pandemic, outlining the key areas for nanoscientists to step in.

In-vitro and in-vivo evaluation of modified sodium starch glycolate for exploring its haemostatic potential.

The control of blood flow from breached blood vessels during surgery or trauma is challenging. With the existing treatment options being either expensive or ineffective, the development of a haemostat that overcome such drawbacks would be beneficial. With an aim to develop an ideal haemostat, the potential of sodium starch glycolate (SSG), a commonly used pharmaceutical disintegrant was modified to obtain porous microparticles (pSSG). The biodegradability, cyto-compatibility and haemo-compatibility of the modified particles were confirmed using appropriate studies. In comparison to starch and SSG, the irregular shaped pSSG demonstrated spontaneous and significant fluid absorption (3500+500 %) and formed a physical barrier to blood flow. In addition, significant blood cells aggregation and platelet activation was observed in the modified micoparticles leading to rapid clot formation. In-vivo studies on liver and abdominal artery injury models in rats indicated the superior haemostatic potential of pSSG over SSG and starch. The results indicated that pSSG can be explored further in clinical evaluation as a hemostat.

In Situ Biosynthesized Superparamagnetic Iron Oxide Nanoparticles (SPIONS) Induce Efficient Hyperthermia in Cancer Cells.

Despite the promising role of magnetic hyperthermia in cancer therapy, its use in patients has been restricted by hurdles that include inefficient targeting of magnetic particles to the tumor site, limited bioavailability, and high toxicity, etc. Taking advantage of the unique metabolic property of cancer cells, we explored the potential of these cells to biosynthesize magnetic nanoparticles for potential hyperthermia applications. Treatment of cancer cells with a mixture of FeCl2 and zinc gluconate resulted in a significant increase in intracellular Fe and Zn content in these cells. Exposure of these cells to an alternating magnetic field (AMF) for 30 min resulted in a substantial temperature rise of 5-6 °C. The in situ formed particles were identified as iron oxide and ZnO nanoparticles. Based on the magnetic property and size, the iron oxide nanoparticles were classified as superparamagnetic iron oxide nanoparticles (SPIONS) comprising a mixture of magnetite (Fe3-δO4) and maghemite (γ-Fe2O3). The role of reactive oxygen species (H2O2) and the involvement of the glycolytic pathway in the biosynthesis of the nanoparticles were confirmed using appropriate in vitro studies. The simplicity of treatment, the specificity of cells capable of synthesis of SPIONS, and the hyperthermia response observed in cancer cells indicate a promising strategy to achieve effective magnetic hyperthermia for cancer therapy.

Polysaccharide-Based Hybrid Self-Healing Hydrogel Supports the Paracrine Response of Mesenchymal Stem Cells.

The aim of stem cell therapy is to repair damaged tissues. Some of the challenges facing its success include cell retention and survival at the wound site. While the retention of cells has been addressed by employing scaffolds, the survival of transplanted cells in the repair tissue is however low. It is hypothesized that the observed regeneration is more a result of migration of tissue repairing cells from adjoining tissues in response to paracrine factors secreted by implanted cells than by the implanted cells per se. In this study, we report the synthesis of a self-healing hybrid hydrogel that is injectable. The hybrid hydrogel was developed using the dynamic equilibrium of Schiff base linkage between the aldehyde groups on carboxymethyl cellulose dialdehyde (CMC-D) and amino groups on carboxymethyl chitosan (CMCh). The hydrogel stiffness and kinetics of gelation were observed to be modulated with different molecular weights of chitosan. In vitro studies demonstrated the cytocompatibility, hemocompatibility, and biodegradability of the hydrogel. The chemotactic, proliferative, and wound-healing response of cells to the paracrine factors secreted from the mesenchymal stem cell (MSC)-hydrogel composite confirmed the ability of the hydrogel to support the paracrine response of stem cells. Our results suggest that the synthesized hydrogel-MSC composite could serve as a potential scaffold for studying the in vitro response of cells to the paracrine factors released by the encapsulated cells as well as a cell delivery vehicle for in vivo applications.

"Gold nanoparticles composite-folic acid conjugated graphene oxide nanohybrids" for targeted chemo-thermal cancer ablation: In vitro screening and in vivo studies.

Nano-graphene oxide (GO) nanometal composite (specifically nanogold and nanosilver) have shown to be a promising material for anticancer therapeutics. Owing to their high drug loading capacity, photothermal and synergizing effects, it is very important to exploit them for targeted chemo-thermal cancer therapeutics. In this work, gold nanoparticles (AuNPs) were selected as the composite metal, folic acid (FA) was taken as GO surface functionalization moiety for active tumor targeting of model anticancer drug Doxorubicin (Dox). AuNPs composite-folate conjugated graphene oxide (FA-GO@Au) nano-platforms were synthesized and characterized in detail. Near-infrared (NIR) sensitivity resulted in an aggravated release of both Dox and ionic gold from the nanohybrid surface. Simultaneous delivery of Dox and AuNPs in the cellular vicinity was further enhanced after localized NIR exposure which resulted in significantly improved cancer cell toxicity. Mechanistic evaluation revealed G0/G1 phase arrest due to increased DNA intercalation and provoked early apoptosis under NIR influence. Pharmacokinetics and organ distribution studies were carried out in healthy mice and rabbits to estimate the actual bio fate of these nanohybrids. In vivo studies showed substantial tumor regression in solid tumor model in Balb/c mice and NIR exposure induced photo-thermal effects further resulted in better tumor management. Study provides substantial evidences both at in vitro and in vivo level to support the fact that NIR induced local photo-thermal effects can solely be used as a tumor targeting tool. This NIR dependent nanohybrid approach presents a precise and flexible strategy for targeted chemotherapy and photo-thermal tumor ablation.