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Bioprinting three-dimensional cell-laden tissue constructs with controllable degradation.

Wu Z, Su X, Xu Y, Kong B, Sun W, Mi S - Sci Rep (2016)

Bottom Line: However, the printed cells in this hydrogel could not degrade the surrounding alginate gel matrix, causing them to remain in a poorly proliferating and non-differentiating state.The 3D-printed hydrogel network with interconnected channels and a macroporous structure was stable and achieved high cell viability (over 90%).Cell proliferation and specific marker protein expression results also revealed that with the help of sodium citrate degradation, the printed HCECs showed a higher proliferation rate and greater cytokeratin 3(CK3) expression, indicating that this newly developed method may help to improve the alginate bioink system for the application of 3D bioprinting in tissue engineering.

View Article: PubMed Central - PubMed

Affiliation: Biomanufacturing Engineering Laboratory, Graduate School at Shenzhen, Tsinghua University, Shenzhen, P.R. China.

ABSTRACT
Alginate hydrogel is a popular biologically inert material that is widely used in 3D bioprinting, especially in extrusion-based printing. However, the printed cells in this hydrogel could not degrade the surrounding alginate gel matrix, causing them to remain in a poorly proliferating and non-differentiating state. Here, we report a novel study of the 3D printing of human corneal epithelial cells (HCECs)/collagen/gelatin/alginate hydrogel incubated with a medium containing sodium citrate to obtain degradation-controllable cell-laden tissue constructs. The 3D-printed hydrogel network with interconnected channels and a macroporous structure was stable and achieved high cell viability (over 90%). By altering the mole ratio of sodium citrate/sodium alginate, the degradation time of the bioprinting constructs can be controlled. Cell proliferation and specific marker protein expression results also revealed that with the help of sodium citrate degradation, the printed HCECs showed a higher proliferation rate and greater cytokeratin 3(CK3) expression, indicating that this newly developed method may help to improve the alginate bioink system for the application of 3D bioprinting in tissue engineering.

No MeSH data available.


Related in: MedlinePlus

Specific marker protein expression of printed cells within constructs incubated with medium containing 66.7% sodium citrate (C/A, mole ratio, %).Micrographs show fluorescent staining of CK3 (green) and nuclei (blue) of HCECs within the constructs after culturing for 1, 3, and 5 days (scale bar, 100 μm). (For the interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
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f8: Specific marker protein expression of printed cells within constructs incubated with medium containing 66.7% sodium citrate (C/A, mole ratio, %).Micrographs show fluorescent staining of CK3 (green) and nuclei (blue) of HCECs within the constructs after culturing for 1, 3, and 5 days (scale bar, 100 μm). (For the interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Mentions: Corneal epithelial-specific cytokeratin 3 (CK3) is often used as a unique marker of HCECs39. Using sodium citrate to accelerate degradation, its expression in HCECs was achieved by immunofluorescence. After 1, 3, and 5 days of culturing in the medium containing 66.7% sodium citrate (C/A, mole ratio, %), the localization and expression of CK3 in HCECs were investigated by confocal microscopy. As Fig. 8 shows, HCECs incubated with sodium citrate proliferated during the culture time. Compared with day 1, the number of cells obviously increased at day 3 or day 5, indicating that the changed matrix property (degradation of the scaffold) may cause a higher cell proliferation, which could be the reason for the increased OD values in the CCK-8 experiment. Additionally, Fig. 8 presents largely synthesised CK3 protein by HCECs printed within the constructs, demonstrating that, using sodium citrate to accelerate the degradation process of the alginate hydrogel system, the cells can not only survive in the hydrogel but also proliferate and synthesise proteins normally. Furthermore, after 3 or 5 days of culturing, it was found that some of the cells formed two- or three-cell aggregates; this was demonstrated more clearly at greater magnification (×200).


Bioprinting three-dimensional cell-laden tissue constructs with controllable degradation.

Wu Z, Su X, Xu Y, Kong B, Sun W, Mi S - Sci Rep (2016)

Specific marker protein expression of printed cells within constructs incubated with medium containing 66.7% sodium citrate (C/A, mole ratio, %).Micrographs show fluorescent staining of CK3 (green) and nuclei (blue) of HCECs within the constructs after culturing for 1, 3, and 5 days (scale bar, 100 μm). (For the interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835808&req=5

f8: Specific marker protein expression of printed cells within constructs incubated with medium containing 66.7% sodium citrate (C/A, mole ratio, %).Micrographs show fluorescent staining of CK3 (green) and nuclei (blue) of HCECs within the constructs after culturing for 1, 3, and 5 days (scale bar, 100 μm). (For the interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)
Mentions: Corneal epithelial-specific cytokeratin 3 (CK3) is often used as a unique marker of HCECs39. Using sodium citrate to accelerate degradation, its expression in HCECs was achieved by immunofluorescence. After 1, 3, and 5 days of culturing in the medium containing 66.7% sodium citrate (C/A, mole ratio, %), the localization and expression of CK3 in HCECs were investigated by confocal microscopy. As Fig. 8 shows, HCECs incubated with sodium citrate proliferated during the culture time. Compared with day 1, the number of cells obviously increased at day 3 or day 5, indicating that the changed matrix property (degradation of the scaffold) may cause a higher cell proliferation, which could be the reason for the increased OD values in the CCK-8 experiment. Additionally, Fig. 8 presents largely synthesised CK3 protein by HCECs printed within the constructs, demonstrating that, using sodium citrate to accelerate the degradation process of the alginate hydrogel system, the cells can not only survive in the hydrogel but also proliferate and synthesise proteins normally. Furthermore, after 3 or 5 days of culturing, it was found that some of the cells formed two- or three-cell aggregates; this was demonstrated more clearly at greater magnification (×200).

Bottom Line: However, the printed cells in this hydrogel could not degrade the surrounding alginate gel matrix, causing them to remain in a poorly proliferating and non-differentiating state.The 3D-printed hydrogel network with interconnected channels and a macroporous structure was stable and achieved high cell viability (over 90%).Cell proliferation and specific marker protein expression results also revealed that with the help of sodium citrate degradation, the printed HCECs showed a higher proliferation rate and greater cytokeratin 3(CK3) expression, indicating that this newly developed method may help to improve the alginate bioink system for the application of 3D bioprinting in tissue engineering.

View Article: PubMed Central - PubMed

Affiliation: Biomanufacturing Engineering Laboratory, Graduate School at Shenzhen, Tsinghua University, Shenzhen, P.R. China.

ABSTRACT
Alginate hydrogel is a popular biologically inert material that is widely used in 3D bioprinting, especially in extrusion-based printing. However, the printed cells in this hydrogel could not degrade the surrounding alginate gel matrix, causing them to remain in a poorly proliferating and non-differentiating state. Here, we report a novel study of the 3D printing of human corneal epithelial cells (HCECs)/collagen/gelatin/alginate hydrogel incubated with a medium containing sodium citrate to obtain degradation-controllable cell-laden tissue constructs. The 3D-printed hydrogel network with interconnected channels and a macroporous structure was stable and achieved high cell viability (over 90%). By altering the mole ratio of sodium citrate/sodium alginate, the degradation time of the bioprinting constructs can be controlled. Cell proliferation and specific marker protein expression results also revealed that with the help of sodium citrate degradation, the printed HCECs showed a higher proliferation rate and greater cytokeratin 3(CK3) expression, indicating that this newly developed method may help to improve the alginate bioink system for the application of 3D bioprinting in tissue engineering.

No MeSH data available.


Related in: MedlinePlus