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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

Proliferation of HCECs within the printed constructs incubated with fresh medium containing no (minus sodium citrate) or 66.7% sodium citrate (plus sodium citrate, mole ratio of C/A), as measured by CCK-8.Data represent means ± SD (*p < 0.01, **p < 0.005).
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f6: Proliferation of HCECs within the printed constructs incubated with fresh medium containing no (minus sodium citrate) or 66.7% sodium citrate (plus sodium citrate, mole ratio of C/A), as measured by CCK-8.Data represent means ± SD (*p < 0.01, **p < 0.005).

Mentions: Moreover, the influence of sodium citrate on the proliferation of HCECs within the printed constructs was measured using CCK-8 (Fig. 6). It was found that, if the cell density was assessed immediately after measuring CCK-8, no difference in the OD was observed between the assays without (0.226) or with (0.236) sodium citrate. However, if sodium citrate was added, the cells showed faster proliferation from day 2 to day 8; on day 8, the OD values were 0.670 ± 0.015 (with sodium citrate) and 0.581 ± 0.021 (without sodium citrate). The former increased 15.4% more than the latter. The morphological changes in the 3D hydrogel during different culture times are shown in Fig. 7. The printed 3D gel constructs were incubated with fresh medium containing 66.7% (C/A, mole ratio, %) sodium citrate. From day 1 to day 10 of the in vitro experiments (Fig. 7), HCECs aggregated and formed clusters, which had a round-shape morphology. The number and diameter of these clusters grew with increasing culture time, which can be clearly shown in Fig. 7. The diameter of the clusters was increased from 14.15 ± 1.77 μm at day 1 to 59.22 ± 3.881 μm at day 10, which was measured using Image Pro Plus (IPP) software.


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)

Proliferation of HCECs within the printed constructs incubated with fresh medium containing no (minus sodium citrate) or 66.7% sodium citrate (plus sodium citrate, mole ratio of C/A), as measured by CCK-8.Data represent means ± SD (*p < 0.01, **p < 0.005).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f6: Proliferation of HCECs within the printed constructs incubated with fresh medium containing no (minus sodium citrate) or 66.7% sodium citrate (plus sodium citrate, mole ratio of C/A), as measured by CCK-8.Data represent means ± SD (*p < 0.01, **p < 0.005).
Mentions: Moreover, the influence of sodium citrate on the proliferation of HCECs within the printed constructs was measured using CCK-8 (Fig. 6). It was found that, if the cell density was assessed immediately after measuring CCK-8, no difference in the OD was observed between the assays without (0.226) or with (0.236) sodium citrate. However, if sodium citrate was added, the cells showed faster proliferation from day 2 to day 8; on day 8, the OD values were 0.670 ± 0.015 (with sodium citrate) and 0.581 ± 0.021 (without sodium citrate). The former increased 15.4% more than the latter. The morphological changes in the 3D hydrogel during different culture times are shown in Fig. 7. The printed 3D gel constructs were incubated with fresh medium containing 66.7% (C/A, mole ratio, %) sodium citrate. From day 1 to day 10 of the in vitro experiments (Fig. 7), HCECs aggregated and formed clusters, which had a round-shape morphology. The number and diameter of these clusters grew with increasing culture time, which can be clearly shown in Fig. 7. The diameter of the clusters was increased from 14.15 ± 1.77 μm at day 1 to 59.22 ± 3.881 μm at day 10, which was measured using Image Pro Plus (IPP) software.

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