Journal: bioRxiv

Article Title: Aqueous two-phase bioinks for discrete packing and compartmentalisation of 3D bioprinted cells

doi: 10.1101/2025.06.27.661968

Figure Lengend Snippet: Rational of ATPS inks bioprinting. (left) The microfluidic printhead is integrated into a 3D bioprinting system to print the ATPS fibres. In the upper inlets, (i) a glycerol solution and (ii) CaCl 2 cross-link solution are used. In the core inlet, ATPS inks based on GelMA 5% and Alginate 4% are flown. (right) The size of the ATPS droplets varies with increasing NaCl and GelMA concentration. (bottom) Within the ATPS hydrogel, cell organisation varies with droplet size, which in turn depends on NaCl concentration. From left to right, cells become increasingly elongated and organised within the GelMA droplets.

Article Snippet: The emulsion ink was then flowed into the microfluidic chip via the control of microfluidic pumps (neMESYS, Cetoni GmbH).

Techniques: Concentration Assay

Journal: bioRxiv

Article Title: Aqueous two-phase bioinks for discrete packing and compartmentalisation of 3D bioprinted cells

doi: 10.1101/2025.06.27.661968

Figure Lengend Snippet: Microfluidic 3D bioprinting of ATPSs. a-i) Image of the microfluidic printhead during 3D deposition of an ATPS scaffold. a-ii) Air-printed ATPS scaffold lattice layer by layer with detail of a single fibre in a-iii) brightfield and in a-iv) alginate autofluorescence at 405 nm. b) OCT images of the ATPS systems. b-i) Printed grid image for the 0 g/L NaCl composition, b-ii) frontal fibre image, b-iii) fibre sections. b-iv) Printed grid image for the 36 g/L NaCl composition, b-v) frontal fibre image, b-vi) fibre sections. Scale bar: (a, iii – iv) 100 µm. Mean ± S.D. n=3.

Article Snippet: The emulsion ink was then flowed into the microfluidic chip via the control of microfluidic pumps (neMESYS, Cetoni GmbH).

Techniques:

Journal: bioRxiv

Article Title: Aqueous two-phase bioinks for discrete packing and compartmentalisation of 3D bioprinted cells

doi: 10.1101/2025.06.27.661968

Figure Lengend Snippet: Spreading and osteogenic differentiation of HBMSCs in ATPS hydrogels at different salt concentrations. a) Microfluidic 3D bioprinting of HBMSCs ATPS-scaffolds. b-c-d) Confocal images with HBMSCs labelled with DAPI to show nuclei (blue) and cells marked with phalloidin to show f-actin (green). e) RT-qPCR analysis of HBMSCs after 7 days of 3D culture in basal and osteogenic media. The protein expression of further factor associated with osteogenic differentiation was examined to observe the effect of different culture media on ATPS scaffolds. The expression levels of osterix (OSX), RUNX2, alkaline phosphatase (ALP), COL1A1, osteonectin (OCL) and osteocalcin (OCN) were found higher in ATPS constructs cultured in osteogenic media at day 7 compared to controls in basal media. f) Immunofluorescence images with HBMSCs labelled with DAPI to show nuclei (blue) and cells marked with BMP-2 monoclonal antibody to show BMP-2 (yellow). g) Microscopic and macroscopic (insert) view of alkaline phosphatase staining performed at day 1 and day 7 in samples at 0 – 9 – 36 g/L NaCl treated in basal and osteogenic medium. Scale bars: (b, f) 100 µm, (g) 100 µm – 1 mm. Statistical significances were assessed by two-way ANOVA. Mean ± S.D. n=3.

Article Snippet: The emulsion ink was then flowed into the microfluidic chip via the control of microfluidic pumps (neMESYS, Cetoni GmbH).

Techniques: Quantitative RT-PCR, Expressing, Construct, Cell Culture, Immunofluorescence, Staining