Localized Delivery of Pilocarpine to Hypofunctional Salivary Glands through Electrospun Nanofiber Mats: An Ex Vivo and In Vivo Study.

Dry mouth or xerostomia is a frequent medical condition among the polymedicated elderly population. Systemic pilocarpine is included in the first line of pharmacological therapies for xerostomia. However, the efficacy of existing pilocarpine formulations is limited due to its adverse side effects and multiple daily dosages. To overcome these drawbacks, a localized formulation of pilocarpine targeting the salivary glands (SG) was developed in the current study. The proposed formulation consisted of pilocarpine-loaded Poly(lactic-co-glycolic acid) (PLGA)/poly(ethylene glycol) (PEG) nanofiber mats via an electrospinning technique. The nanofiber mats were fully characterized for their size, mesh porosity, drug encapsulation efficiency, and in vitro drug release. Mat biocompatibility and efficacy was evaluated in the SG organ ex vivo, and the expression of proliferation and pro-apoptotic markers at the cellular level was determined. In vivo short-term studies were performed to evaluate the saliva secretion after acute SG treatment with pilocarpine-loaded nanofiber mats, and after systemic pilocarpine for comparison purposes. The outcomes demonstrated that the pilocarpine-loaded mats were uniformly distributed (diameter: 384 ± 124 nm) in a highly porous mesh, and possessed a high encapsulation efficiency (~81%). Drug release studies showed an initial pilocarpine release of 26% (4.5 h), followed by a gradual increase (~46%) over 15 d. Pilocarpine-loaded nanofiber mats supported SG growth with negligible cytotoxicity and normal cellular proliferation and homeostasis. Salivary secretion was significantly increased 4.5 h after intradermal SG treatment with drug-loaded nanofibers in vivo. Overall, this study highlights the strengths of PLGA/PEG nanofiber mats for the localized daily delivery of pilocarpine and reveals its potential for future clinical translation in patients with xerostomia.

3D bioprinting of skin tissue: From pre-processing to final product evaluation.

Bioprinted skin tissue has the potential for aiding drug screening, formulation development, clinical transplantation, chemical and cosmetic testing, as well as basic research. Limitations of conventional skin tissue engineering approaches have driven the development of biomimetic skin equivalent via 3D bioprinting. A key hope for bioprinting skin is the improved tissue authenticity over conventional skin equivalent construction, enabling the precise localization of multiple cell types and appendages within a construct. The printing of skin faces challenges broadly associated with general 3D bioprinting, including the selection of cell types and biomaterials, and additionally requires in vitro culture formats that allow for growth at an air-liquid interface. This paper provides a thorough review of current 3D bioprinting technologies used to engineer human skin constructs and presents the overall pipelines of designing a biomimetic artificial skin via 3D bioprinting from the design phase (i.e. pre-processing phase) through the tissue maturation phase (i.e. post-processing) and into final product evaluation for drug screening, development, and drug delivery applications.

Evaluation of sewage sludge incineration ash as a potential land reclamation material.

This study evaluated the potential of utilising sewage sludge incineration ash as a land reclamation material. Toxicity assessment of the leachate of the ash was carried out for both terrestrial and marine organisms. Both the fruit fly Drosophila melanogaster and barnacle Amphibalanus amphitrite showed that both bottom and fly ash leached at liquid-to-solid (L/S) ratio 5 did not substantially affect viabilities. The leachate carried out at L/S 10 was compared to the European Waste Acceptance Criteria and the sewage sludge ashes could be classified as non-hazardous waste. The geotechnical properties of the sewage sludge ash were studied and compared to sand, a conventional land reclamation material, for further evaluation of its potential as a land reclamation material. It was found from direct shear test that both bottom and fly ashes displayed similar and comparable shear strength to that of typical compacted sandy soil based on the range of internal friction angle obtained. However, the consolidation profile of bottom ash was significantly different from sand, while that of fly ash was more similar to sand. Our study showed that the sewage sludge ash has the potential to be used as a land reclamation material.

Effective Recovery of Vanadium from Oil Refinery Waste into Vanadium-Based Metal-Organic Frameworks.

Carbon black waste, an oil refinery waste, contains a high concentration of vanadium(V) leftover from the processing of crude oil. For the sake of environmental sustainability, it is therefore of interest to recover the vanadium as useful products instead of disposing of it. In this work, V was recovered in the form of vanadium-based metal-organic frameworks (V-MOFs) via a novel pathway by using the leaching solution of carbon black waste instead of commercially available vanadium chemicals. Two different types of V-MOFs with high levels of crystallinity and phase purity were fabricated in very high yields (>98%) based on a coordination modulation method. The V-MOFs exhibited well-defined and controlled shapes such as nanofibers (length: > 10 µm) and nanorods (length: ∼270 nm). Furthermore, the V-MOFs showed high catalytic activities for the oxidation of benzyl alcohol to benzaldehyde, indicating the strong potential of the waste-derived V-MOFs in catalysis applications. Overall, our work offers a green synthesis pathway for the preparation of V-MOFs by using heavy metals of industrial waste as the metal source.

Codelivery of anti-cancer agents via double-walled polymeric microparticles/injectable hydrogel: A promising approach for treatment of triple negative breast cancer.

Triple negative breast cancer (TNBC) is an aggressive sub-type of breast cancer that rarely responds to conventional chemotherapy. Therefore, novel agents or new routes need to be developed to improve treatment efficacy and diminish severe side-effects of anti-cancer agents in TNBC patients. This study explores a novel localized co-delivery platform with potential application against TNBC. Uniform core-shell microparticles encapsulating cisplatin (Cis-DDP) and paclitaxel (PTX) are fabricated using coaxial electrohydrodynamic atomization technique and subsequently are embedded into an injectable hydrogel. The hydrogel provides an additional diffusion barrier against Cis-DDP and confines premature release of drugs. In addition, the hydrogel can provide a versatile tool for retaining particles in the tumor resected cavity during the injection following debulking surgery and prevent surgical site infection due to its inherent antibacterial properties. The combination of Cis-DDP and PTX demonstrates a synergistic effect against MDA-MB-231 cell line assigned to three different mechanisms of action, including denaturation of DNA strands, stabilization of microtubules, and amplification of intracellular reactive oxygen species (ROS) and activation of caspase-3 pathways. The results show a significant accumulation of mitochondrial ROS insults in cells upon treatment that consequently causes programmed cells death. The performance of microparticles/hydrogel carrier is evaluated against three-dimensional MDA-MB-231 (breast cancer) 3D spheroids, where a superior efficacy and a greater reduction in spheroid growth are observed over 14 days, as compared with free-drug treatment. Overall, drug-loaded core-shell microparticles embedded into injectable hydrogel provides a promising strategy to treat aggressive cancers and a modular platform for a broad range of localized multidrug therapies customizable to the cancer type.

Chemically treated carbon black waste and its potential applications.

In this work, carbon black waste - a hazardous solid residue generated from gasification of crude oil bottom in refineries - was successfully used for making an absorbent material. However, since the carbon black waste also contains significant amounts of heavy metals (especially nickel and vanadium), chemical leaching was first used to remove these hazardous impurities from the carbon black waste. Acid leaching with nitric acid was found to be a very effective method for removal of both nickel and vanadium from the carbon black waste (i.e. up to 95% nickel and 98% vanadium were removed via treatment with 2M nitric acid for 1h at 20°C), whereas alkali leaching by using NaOH under the same condition was not effective for removal of nickel (less than 10% nickel was removed). Human lung cells (MRC-5) were then used to investigate the toxicity of the carbon black waste before and after leaching. Cell viability analysis showed that the leachate from the original carbon black waste has very high toxicity, whereas the leachate from the treated samples has no significant toxicity. Finally, the efficacy of the carbon black waste treated with HNO3 as an absorbent for dye removal was investigated. This treated carbon black waste has high adsorption capacity (∼361.2mg dye/g carbonblack), which can be attributed to its high specific surface area (∼559m2/g). The treated carbon black waste with its high adsorption capacity and lack of cytotoxicity is a promising adsorbent material. Moreover, the carbon black waste was found to show high electrical conductivity (ca. 10S/cm), making it a potentially valuable source of conductive material.

Toxicity assessment of carbon black waste: A by-product from oil refineries.

In Singapore, approximately 30t/day of carbon-based solid waste are produced from petrochemical processes. This carbon black waste has been shown to possess physical properties that are characteristic of a good adsorbent such as high external surface area. Therefore, there is a growing interest to reutilize and process this carbon black waste into secondary materials such as adsorbents. However, the carbon black waste obtained from petrochemical industries may contain heavy metals that are hazardous to human health and the environment, hence restricting its full potential for re-utilization. Therefore, it is important to examine the possible toxicity effects and toxicity mechanism of carbon black waste on human health. In this study, inductively coupled plasma optical emission spectroscopy (ICP-OES) analysis showed that the heavy metals, vanadium (V), molybdenum (Mo) and nickel (Ni), were present in the carbon black waste in high concentrations. Three human cell lines (HepG2 cells, MRC-5 cells and MDA-MB-231 cells) were used to investigate the toxicity of carbon black waste extract in a variety of in vitro assays. Results from MTS assays indicated that carbon black waste extract decreased the viability of all three cell lines in a dose and time-dependent manner. Observations from confocal microscopy further confirmed this phenomenon. Flow cytometry assay also showed that carbon black waste extract induced apoptosis of human cell lines, and the level of apoptosis increased with increasing waste concentration. Results from reactive oxygen species (ROS) assay indicated that carbon black waste extract induced oxidative stress by increasing intracellular ROS generation in these three human cell lines. Moreover, induction of oxidative damage in these cells was also observed through the alteration of glutathione (GSH) and superoxide dismutase (SOD) activities. Last but not least, by treating the cells with V-spiked solution of concentration equivalent to that found in the carbon black waste extract, V was identified as the main culprit for the high toxicity of carbon black waste extract. These findings could potentially provide insight into the hazards of carbon black waste extract and its toxicity mechanism on human cell lines.

Double-Walled Microparticles-Embedded Self-Cross-Linked, Injectable, and Antibacterial Hydrogel for Controlled and Sustained Release of Chemotherapeutic Agents.

First-line cancer chemotherapy has been prescribed for patients suffered from cancers for many years. However, conventional chemotherapy provides a high parenteral dosage of anticancer drugs over a short period, which may cause serious toxicities and detrimental side effects in healthy tissues. This study aims to develop a new drug delivery system (DDS) composed of double-walled microparticles and an injectable hydrogel for localized dual-agent drug delivery to tumors. The uniform double-walled microparticles loaded with cisplatin (Cis-DDP) and paclitaxel (PTX) were fabricated via coaxial electrohydrodynamic atomization (CEHDA) technique and subsequently were embedded into injectable alginate-branched polyethylenimine. The findings show the uniqueness of CEHDA technique for simply swapping the place of drugs to achieve a parallel or a sequential release profile. This study also presents the simulation of CEHDA technique using computational fluid dynamics (CFD) that will help in the optimization of CEHDA's operating conditions prior to large-scale production of microparticles. The new synthetic hydrogel provides an additional diffusion barrier against Cis-DDP and confines premature release of drugs. In addition, the hydrogel can provide a versatile tool for retaining particles in the tumor resected cavity during the injection after debulking surgery and preventing surgical site infection due to its inherent antibacterial properties. Three-dimensional MDA-MB-231 (breast cancer) spheroid studies demonstrate a superior efficacy and a greater reduction in spheroid growth for drugs released from the proposed composite formulation over a prolonged period, as compared with free drug treatment. Overall, the new core-shell microparticles embedded into injectable hydrogel can serve as a flexible controlled release platform for modulating the release profiles of anticancer drugs and subsequently providing a superior anticancer response.

Rapid toxicity screening of gasification ashes.

The solid residues including bottom ashes and fly ashes produced by waste gasification technology could be reused as secondary raw materials. However, the applications and utilizations of these ashes are very often restricted by their toxicity. Therefore, toxicity screening of ash is the primary condition for reusing the ash. In this manuscript, we establish a standard for rapid screening of gasification ashes on the basis of in vitro and in vivo testing, and henceforth guide the proper disposal of the ashes. We used three different test models comprising human cell lines (liver and lung cells), Drosophila melanogaster and Daphnia magna to examine the toxicity of six different types of ashes. For each ash, different leachate concentrations were used to examine the toxicity, with C0 being the original extracted leachate concentration, while C/C0 being subsequent diluted concentrations. The IC50 for each leachate was also quantified for use as an index to classify toxicity levels. The results demonstrated that the toxicity evaluation of different types of ashes using different models is consistent with each other. As the different models show consistent qualitative results, we chose one or two of the models (liver cells or lung cells models) as the standard for rapid toxicity screening of gasification ashes. We may classify the gasification ashes into three categories according to the IC50, 24h value on liver cells or lung cells models, namely "toxic level I" (IC50, 24h>C/C0=0.5), "toxic level II" (C/C0=0.05

Potential application of gasification to recycle food waste and rehabilitate acidic soil from secondary forests on degraded land in Southeast Asia.

Gasification is recognized as a green technology as it can harness energy from biomass in the form of syngas without causing severe environmental impacts, yet producing valuable solid residues that can be utilized in other applications. In this study, the feasibility of co-gasification of woody biomass and food waste in different proportions was investigated using a fixed-bed downdraft gasifier. Subsequently, the capability of biochar derived from gasification of woody biomass in the rehabilitation of soil from tropical secondary forests on degraded land (adinandra belukar) was also explored through a water spinach cultivation study using soil-biochar mixtures of different ratios. Gasification of a 60:40 wood waste-food waste mixture (w/w) produced syngas with the highest lower heating value (LHV) 5.29 MJ/m(3)-approximately 0.4-4.0% higher than gasification of 70:30 or 80:20 mixtures, or pure wood waste. Meanwhile, water spinach cultivated in a 2:1 soil-biochar mixture exhibited the best growth performance in terms of height (a 4-fold increment), weight (a 10-fold increment) and leaf surface area (a 5-fold increment) after 8 weeks of cultivation, owing to the high porosity, surface area, nutrient content and alkalinity of biochar. It is concluded that gasification may be an alternative technology to food waste disposal through co-gasification with woody biomass, and that gasification derived biochar is suitable for use as an amendment for the nutrient-poor, acidic soil of adinandra belukar.