Applications of alginate microspheres in therapeutics delivery and cell culture: Past, present and future

Polymers are the backbone of pharmaceutical drug delivery. There are several polymers with varying properties available today for use in different pharmaceutical applications. Alginate is widely used in biomedical research due to its attractive features such as biocompatibility, biodegradability, inertness, low cost, and ease of production and formulation. Encapsulation of therapeutic agents in alginate/alginate complex microspheres protects them from environmental stresses, including the acidic environment in the gastro-intestinal tract (GIT) and enzymatic degradation, and allows targeted and sustained delivery of the agents. Microencapsulation is playing an increasingly important role in drug delivery as evidenced by the recent surge in research articles on the use of alginate in the delivery of small molecules, cells, bacteria, proteins, vaccines, and for tissue engineering applications. Formulation of these alginate microspheres (AMS) are commonly achieved by conventional external gelation method using various instrumental manipulation such as vortexing, homogenization, ultrasonication or spray drying, and each method affects the overall particle characteristics. In this review, an inclusive summary of the currently available methods for the formulation of AMS, its recent use in the encapsulation and delivery of therapeutics, and future outlook will be discussed

Fabrication of PNIPAm-based thermoresponsive hydrogel microwell arrays for tumor spheroid formation

Complex three-dimensional (3D) cell cultures are being increasingly implemented in biomedical research as they provide important insights into complex cancer biology, and cell-cell and cell-matrix interactions in the tumor microenvironment. However, most methods used today for 3D cell culture are limited by high cost, the need for specialized skills, low throughput and the use of unnatural culture environments. We report the development of a unique biomimetic hydrogel microwell array platform for the generation and stress-free isolation of cancer spheroids. The poly N-isopropylacrylamide-based hydrogel microwell array (PHMA) has thermoresponsive properties allowing for the attachment and growth of cell aggregates/ spheroids at 37 °C, and their easy isolation at room temperature (RT). The reversible phase transition of the microwell arrays at 35 °C was confirmed visually and by differential scanning calorimetry. Swelling/ shrinking studies and EVOS imaging established that the microwell arrays are hydrophilic and swollen at temperatures \u3c35 °C, while they shrink and are hydrophobic at temperatures \u3e35 °C. Spheroid development within the PHMA was optimized for seeding density, incubation time and cell viability. Spheroids of A549, HeLa and MG-63 cancer cell lines, and human lung fibroblast (HLF) cell line generated within the PHMAs had relatively spherical morphology with hypoxic cores. Finally, using MG-63 cell spheroids as representative models, a proof-of-concept drug response study using doxorubicin hydrochloride was conducted. Overall, we demonstrate that the PHMAs are an innovative alternative to currently used 3D cell culture techniques, for the high-throughput generation of cell spheroids for disease modeling and drug screening applications

Emerging drug delivery strategies for idiopathic pulmonary fibrosis treatment

Idiopathic pulmonary fibrosis (IPF) is a debilitating and fatal condition that causes severe scarring of the lungs. While the pathogenesis of IPF continues to be extensively studied and several factors have been considered, an exact cause has yet to be established. With inadequate treatment options and no cure available, overall disease prognosis is still poor. Existing oral therapies, pirfenidone and nintedanib, may attempt to improve the patients’ quality of life by mitigating symptoms and slowing disease progression, however chronic doses and systemic deliveries of these drugs can lead to severe side effects. The lack of effective treatment options calls for further investigation of restorative as well as additional palliative therapies for IPF. Nanoparticle-based sustained drug delivery strategies can be utilized to ensure targeted delivery for site-specific treatment as well as long-acting therapy, improving overall patient compliance. This review provides an update on promising strategies for the delivery of anti-fibrotic agents, along with an overview of key therapeutic targets as well as relevant emerging therapies currently being evaluated for IPF treatment

Dual-Modality, Dual-Functional Nanoprobes for Cellular and Molecular Imaging

<p>An emerging need for evaluation of promising cellular therapies is a non-invasive method to image the movement and health of cells following transplantation. However, the use of a single modality to serve this purpose may not be advantageous as it may convey inaccurate or insufficient information. Multi-modal imaging strategies are becoming more popular for <i>in vivo</i> cellular and molecular imaging because of their improved sensitivity, higher resolution and structural/functional visualization. This study aims at formulating Nile Red doped hexamethyldisiloxane (HMDSO) nanoemulsions as dual modality (Magnetic Resonance Imaging/Fluorescence), dual-functional (oximetry/detection) nanoprobes for cellular and molecular imaging. HMDSO nanoprobes were prepared using a HS15-lecithin combination as surfactant and showed an average radius of 71&#177;39 nm by dynamic light scattering and <i>in vitro</i> particle stability in human plasma over 24 hrs. They were found to readily localize in the cytosol of MCF7-GFP cells within 18 minutes of incubation. As proof of principle, these nanoprobes were successfully used for fluorescence imaging and for measuring pO₂ changes in cells by magnetic resonance imaging, <i>in vitro,</i> thus showing potential for <i>in vivo</i> applications.</p

Biodegradable and Inherently Fluorescent pH-Responsive Nanoparticles for Cancer Drug Delivery

Purpose: The development of two novel pH-only and pH- and thermo-responsive theranostic nanoparticle (NP) formulations to deliver an anticancer drug and track the accumulation and therapeutic efficacy of the formulations through inherent fluorescence. Methods: A pH-responsive formulation was synthesized from biodegradable photoluminescent polymer (BPLP) and sodium bicarbonate (SBC) via an emulsion technique, while a thermoresponsive BPLP copolymer (TFP) and SBC were used to synthesize a dual-stimuli responsive formulation via free radical co-polymerization. Cisplatin was employed as a model drug and encapsulated during synthesis. Size, surface charge, morphology, pH-dependent fluorescence, lower critical solution temperature (LCST; TFP NPs only), cytocompatibility and in vitro uptake, drug release kinetics and anticancer efficacy were assessed. Results: While all BPLP-SBC and TFP-SBC combinations produced spherical nanoparticles of a size between 200-300 nm, optimal polymer-SBC ratios were selected for further study. Of these, the optimal BPLP-SBC formulation was found to be cytocompatible against primary Type-1 alveolar epithelial cells (AT1) up to 100 μg/mL, and demonstrated sustained drug release over 14 days, dose-dependent uptake, and marked pH-dependent A549 cancer cell killing (72 vs. 24% cell viability, at pH 7.4 vs. 6.0). The optimal TFP-SBC formulation showed excellent cytocompatibility against AT1 cells up to 500 μg/mL, sustained release characteristics, dose-dependent uptake, pH-dependent (78% at pH 7.4 vs. 64% at pH 6.0 at 37°C) and marked temperature-dependent A549 cancer cell killing (64% at 37°C vs. 37% viability at pH 6.0, 41°C). Conclusions: In all, both formulations hold promise as inherently fluorescent, stimuli-responsive theranostic platforms for passively targeted anti-cancer therapy. Graphical Abstract: [Figure not available: see fulltext.]

Synthesis and characterization of a novel pH-responsive drug-releasing nanocomposite hydrogel for skin cancer therapy and wound healing

Local skin cancer recurrence occurs in ∼12% of the patients post-surgery due to persistent growth of residual cancer cells. Wound infection is another significant complication following surgery. We report a novel in situ-forming nanocomposite hydrogel (NCH) containing PLGA-carboxymethyl chitosan nanoparticles (186 nm) for localized pH-responsive skin cancer therapy and wound healing. This injectable hydrogel, comprising of a citric acid-derived polymer backbone, gelled within 5 minutes, and demonstrated excellent swelling (283% of dry weight) and compressive strengths (∼5.34 MPa). Nanoparticle incorporation did not significantly affect hydrogel properties. The NCH effluents were cytocompatible with human dermal fibroblasts at 500 μg ml-1 concentration and demonstrated pH-dependent drug release and promising therapeutic efficacy against A431 and G361 skin cancer cells in vitro. Significant zones of inhibition were observed in S. aureus and E. coli cultures on NCH treatment, confirming its antibacterial properties. Our studies show that the pH-responsive NCH can be potentially used for adjuvant skin cancer treatment and wound healing. This journal i

Magnetic hydroxyapatite nanomaterial–cyclodextrin tethered polymer hybrids as anticancer drug carriers

Osteosarcoma, the most common bone cancer, leads to a poor survival rate of patients. Drug targeting employing hydroxyapatite (HAp)-based nanocarriers represents a fascinating choice for non-invasive treatment of osteosarcoma. Herein, we report strontium-doped (Sr-HAp) and iron- and strontium-co-doped (Sr,Fe-HAp) hydroxyapatite nanoparticles as novel materials that deliver doxorubicin to bone cancer cells. A platinum-complexed and cyclodextrin-functionalized chitosan derivative is utilized to coat the NPs. Sr-HAp (aspect ratio ∼20) and Sr,Fe-HAp (aspect ratio ∼3) nanoparticles are formed as nanowhiskers and nanorods, respectively, as revealed by transmission electron microscopy. Strontium ferrite NPs are synthesized and their properties are compared with those of the Sr/Sr,Fe-doped HAp NPs. These ferrite NPs show ferromagnetic behavior, as opposed to Sr-HAp and Sr,Fe-HAp. The latter two respectively display paramagnetic and superparamagnetic behaviors. The loading percentage of the anticancer drug, Doxorubicin (Dox), in the nanocarriers is high and the release of Dox is sustained at physiological pH. The Dox-loaded nanocarriers are tested for their in vitro cytotoxicity against lung, cervical, liver, and bone cancer cell lines. In general, the efficacy of Dox is not diminished on loading in the nanocarriers. In addition, the Dox-carriers demonstrate a time- and dose-dependent cytotoxicity. The efficacy is enhanced in the case of Dox-loaded carriers on MG-63 (osteosarcoma) cell lines. The anticancer activity is tested in vivo on both male and female albino mice. Enhanced chemotherapeutic potential is observed for Dox-loaded Sr,Fe-HAp in a metastatic model of MG-63. The platinum derivative polymer possesses its own therapeutic effect and contributes to the general activity. The novel polymer-HAp nanohybrid represents an effective nanocarrier for the treatment of osteosarcoma