Nanofibrous polyelectrolyte complex incorporated BSA-alginate composite bioink for 3D bioprinting of bone mimicking constructs.

Although several bioinks have been developed for 3D bioprinting applications, the lack of optimal printability, mechanical properties, and adequate cell response has limited their practical applicability. Therefore, this work reports the development of a composite bioink consisting of bovine serum albumin (BSA), alginate, and self-assembled nanofibrous polyelectrolyte complex aggregates of gelatin and chitosan (PEC-GC). The nanofibrous PEC-GC aggregates were prepared and incorporated into the bioink in varying concentrations (0 % to 3 %). The bioink samples were bioprinted and crosslinked post-printing by calcium chloride. The average nanofiber diameter of PEC-GC was 62 ± 15 nm. It was demonstrated that PEC-GC improves the printability and cellular adhesion of the developed bioink and modulates the swelling ratio, degradation rate, and mechanical properties of the fabricated scaffold. The in vitro results revealed that the bioink with 2 % PEC-GC had the best post-printing cell viability of the encapsulated MG63 osteosarcoma cells and well oragnized stress fibers, indicating enhanced cell adhesion. The cell viability was >90 %, as observed from the MTT assay. The composite bioink also showed osteogenic potential, as confirmed by the estimation of alkaline phosphatase activity and collagen synthesis assay. This study successfully fabricated a high-shape fidelity bioink with potential in bone tissue engineering.

Development of a novel thermogelling PEC-based ECM mimicking nanocomposite bioink for bone tissue engineering.

Non-union of large bone defects has been an existing clinical problem. 3D extrusion-based bioprinting provides an efficient approach to tackle such problems. This approach enables the use of various biomaterials, cell types and growth factors in developing a superior bone graft that is specific to the defect. In this article, we have designed and printed an ECM mimicking, self-assembled polyelectrolyte complex (PEC) based fibrous bioink using natural polymers like chitosan-polygalacturonic acid (PGA) and other biomaterials - gelatin, laponite and nanohydroxyapatite with a modified 3D printer. The developed bioink possesses a thermo-reversible sol-gel transition at physiological pH and temperature. Here, we demonstrated that post-printing, our fiber-reinforced bioink had significant cell proliferation with cell viability of >80% and negligible cell morbidity. The practicability of developing this self-assembled PEC-based bioink was assessed. Bioink with 4% gelatin (PECHLG4) had optimal printability with a minimal swelling ratio of approximately 3%. The printed scaffold had integrity for a period of 8 days under 0.5 mg/mL lysozyme concentration. We also evaluated the mechanical property of the bioink using compression analysis which gave an elastic modulus of 16 KPa. This combination of natural polymers and nanocomposite, along with a fibrous network of PECs, is itself a novel approach for 3D bioprinting and can be a preliminary proposition for the treatment of large bone defects.

Valorization of phenol contaminated wastewater for lipid production by Rhodosporidium toruloides 9564T.

Phenol is one of the most common hazardous organic compound presents in several industrial effluents which directly affects the aquatic environment. The present study envisaged the phenol biodegradation and simultaneous lipid production along with its underlying mechanism by oleaginous yeast Rhodosporidium toruloides 9564T. Experiments were designed using simulated wastewater by varying phenol concentration in the range of 0.25-1.5 g/L and inoculum size of 1, 5, and 10% with and without glucose. The oleaginous yeast was found to completely degrade up to 0.75 g/L phenol with lipid accumulation of 26.3%. Phenol at > 0.5 g/L severely inhibited the growth of R. toruloides 9564T at 1% and 5% inoculum size. Phenol toxicity up to 0.75 g/L can be overcome by increasing inoculum size to 10%. The maximum specific growth rate (µmax) and phenol degradation rate (qmax) were found to be 0.0717 h-1 and 0.01523 h-1, respectively. The enzymatic pathway study suggested that R. toruloides 9564T follows an ortho cleavage pathway for phenol degradation and lipid accumulation. Phytotoxicty and cytotoxicity tests for treated and untreated samples clearly demonstrated a decline in toxicity of the treated wastewater. R. toruloides brought about an important paradigm shift toward a circular economy in which industrial wastewater is considered a valuable resource for bioenergy production.