Charge-Controlled Synthetic Hyaluronan-Based Cell Matrices
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthetic ECM Model with Tunable Charge
2.2. Charge-Dependent Enzymatic Degradation
2.2.1. Hyaluronidase IV
2.2.2. Hyaluronate Lyase
2.2.3. Comparison
2.3. Charge-Dependent Protein Adsorption
2.3.1. Unspecific
2.3.2. Specific
2.4. Charge-Dependent Cell Attachment
3. Conclusions
4. Materials and Methods
4.1. Materials
4.2. HA Hydrogel Formation and Physicochemical Characterization
4.2.1. HA Thiolation
4.2.2. HA Hydrogel Formation and Determination of Reacted Thiols
4.2.3. Mechanical Measurements
4.3. Calcultation of Negative Network Charge
4.4. Enzymatic Degradation
4.5. Protein Adsorption
4.6. Cell Cultivation on HA Hydrogels
4.7. Phase Contrast and Fluorescence Microscopy on HA Hydrogels
4.7.1. Live/Dead Staining
4.7.2. Microscopy Imaging
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Sample Availability: Samples of the compounds are available from the authors. |
Degree of Thiolation (%) | Adsorbed Collagen (ng/mL) | Adsorbed Fibronectin (ng/mL) | Adsorbed Aggrecan (ng/mL) |
---|---|---|---|
18 ± 1 | 0.001 ± 0.019 | −0.020 ± 0.011 | −0.011 ± 0.019 |
25 ± 4 | −0.015 ± 0.004 | −0.029 ± 0.006 | −0.024 ± 0.007 |
33 ± 2 | −0.020 ± 0.004 | −0.023 ± 0.012 | −0.031 ± 0.004 |
Ratio of HA:DTP:EDCl | Batch of HA (Lot Lifecore) | Reaction Time | Degree of Thiolation |
---|---|---|---|
1:1:1 | 025828 | 10 min | 18 ± 1% |
1:1:1 | 025828 | 15 min | 25 ± 1% |
1:1:1 | 025828 | 20 min | 33 ± 2% |
Degree of Thiolation (%) | Crosslinker | Young’s Modulus (kPa) |
---|---|---|
18 ± 1 | uncharged | 0.36 ± 0.02 |
18 ± 1 | charged | 2.57 ± 0.26 |
25 ± 4 | uncharged | 2.68 ± 0.13 |
25 ± 4 | charged | 3.65 ± 0.71 |
33 ± 2 | uncharged | 3.20 ± 0.12 |
33 ± 2 | charged | 6.57 ± 0.71 |
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Hegger, P.S.; Kupka, J.; Minsky, B.B.; Laschat, S.; Boehm, H. Charge-Controlled Synthetic Hyaluronan-Based Cell Matrices. Molecules 2018, 23, 769. https://doi.org/10.3390/molecules23040769
Hegger PS, Kupka J, Minsky BB, Laschat S, Boehm H. Charge-Controlled Synthetic Hyaluronan-Based Cell Matrices. Molecules. 2018; 23(4):769. https://doi.org/10.3390/molecules23040769
Chicago/Turabian StyleHegger, Patricia S., Julia Kupka, Burcu Baykal Minsky, Sabine Laschat, and Heike Boehm. 2018. "Charge-Controlled Synthetic Hyaluronan-Based Cell Matrices" Molecules 23, no. 4: 769. https://doi.org/10.3390/molecules23040769