Alginate-based hydrogel promotes neuronal survival and axon outgrowth of neuron-like cells
Year 2023,
Volume: 40 Issue: 2, 219 - 224, 19.07.2023
Başak Dalbayrak
,
Ekin Sönmez
,
Habibe Kurt
,
Müge İşleten Hoşoğlu
,
N. Hale Saybasili
,
Işıl Aksan Kurnaz
Abstract
Alginate is a natural polymer preferred for biotechnological applications due to its properties, such as biocompatibility, biodegradability, and low toxicity. However, neurons do not possess surface molecules interacting with alginate; therefore, alginate-based materials have limitations for neurodegenerative applications. Thus, increasing neuronal survival and promoting axonal outgrowth in the alginate-based hydrogels are the primary purposes of this study. We also aim to study the performance of alginate extracted from bioresources to that of commercial alginate. Cell-embedded alginate-based hydrogels were formed with CaCl2 and were either mixed with collagen type I or supplemented with differentiation protocols such as the addition of growth factors NGF or FGF, as well as serum withdrawal and retinoic acid (RA). Cells were observed by fluorescence imaging with acridine orange and propidium iodide, and upon dissolving the hydrogel with EDTA, cells were counted with trypan blue staining. In this study, both commercial alginate as well as alginate extracted from seaweed were compared for their performance and were found to be comparable. We determined that the addition of collagen to the alginate hydrogel increased neuronal survival but not axon outgrowth. NSC-34 cell differentiation with NGF and FGF was successful in both commercial and extracted alginate, with both growth factors increasing neural survival as well as axonal outgrowth, in spite of the clustering of cells immediately after treatment. However, the SH-SY5Y differentiation protocol using serum withdrawal and RA treatment did not yield good results. Both extracted and commercial alginates showed comparable performance in terms of neuronal survival in our study, which was further increased upon collagen addition. We also showed that NGF and FGF differentiation protocol in alginate hydrogels resulted in successful axon outgrowth in NSC-34 cells.
Thanks
The alginate extraction was performed within the scope of the OBEK project funded by Dogu Marmara Development Agency (MARKA), and the commercial alginate was a gift from Assoc. Prof. Israfil Kucuk at Gebze Technical University. We would like to acknowledge Prof. Dr. E. Damla Arisan for valuable help and supervision in this manuscript's critical writing and editing.
References
- 1. Hoffman AS. Hydrogels for biomedical applications. Adv Drug Deliv Rev [Internet]. 2012;64(SUPPL.):18–23. Available from: http://dx.doi.org/10.1016/j.addr.2012.09.010
- 2. Ahearne M, Ahearne M. Introduction to cell – hydrogel mechanosensing. 2014;
- 3. Oyen ML. Mechanical characterisation of hydrogel materials. Int Mater Rev. 2014;59(1):44–59.
- 4. Ozcelik B. Degradable hydrogel systems for biomedical applications [Internet]. Biosynthetic Polymers for Medical Applications. Elsevier Ltd; 2016. 173–188 p. Available from: http://dx.doi.org/10.1016/B978-1-78242-105-4.00007-9
- 5. Draget KI. Alginates. Handb Hydrocoll Second Ed. 2009;807–28.
- 6. Paques JP. Alginate Nanospheres Prepared by Internal or External Gelation with Nanoparticles [Internet]. Microencapsulation and Microspheres for Food Applications. Elsevier Inc.; 2015. 39–55 p. Available from: http://dx.doi.org/10.1016/B978-0-12-800350-3.00004-2
- 7. Rowley JA, Madlambayan G, Mooney DJ. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials. 1999;20(1):45–53.
- 8. Wang B, Wan Y, Zheng Y, Lee X, Liu T, Yu Z, et al. Alginate-based composites for environmental applications: a critical review. Crit Rev Environ Sci Technol [Internet]. 2019;49(4):318–56. Available from: https://doi.org/10.1080/10643389.2018.1547621
- 9. Andersen T, Auk-Emblem P, Dornish M. 3D Cell Culture in Alginate Hydrogels. Microarrays. 2015;4(2):133–61.
- 10. Kular JK, Basu S, Sharma RI. The extracellular matrix: Structure, composition, age-related differences, tools for analysis and applications for tissue engineering. J Tissue Eng. 2014;5.
- 11. Dalbayrak, Basak, Sonmez E, Kurt H, Isleten Hosoglu M, Kuçuk I, et al. Characterization of a 3D Neuronal-Culture Using Alginate Hydrogels and Optimize For Neuronal Survival and Axon Growth. Nat Appl Sci J. 2021;3(Special).
- 12. Kandemir B, Caglayan B, Hausott B, Erdogan B, Dag U, Demir O, et al. Pea3 transcription factor promotes neurite outgrowth. Front Mol Neurosci. 2014;7(JUNE):1–11.
- 13. Wang H, Wang R, Thrimawithana T, Little PJ, Xu J, Feng ZP, et al. The Nerve Growth Factor Signaling and Its Potential as Therapeutic Target for Glaucoma. Biomed Res Int. 2014;2014.
- 14. Bradshaw RA, Mobley W, Rush RA. Nerve growth factor and related substances: A brief history and an introduction to the international NGF meeting series. Int J Mol Sci. 2017;18(6).
Year 2023,
Volume: 40 Issue: 2, 219 - 224, 19.07.2023
Başak Dalbayrak
,
Ekin Sönmez
,
Habibe Kurt
,
Müge İşleten Hoşoğlu
,
N. Hale Saybasili
,
Işıl Aksan Kurnaz
References
- 1. Hoffman AS. Hydrogels for biomedical applications. Adv Drug Deliv Rev [Internet]. 2012;64(SUPPL.):18–23. Available from: http://dx.doi.org/10.1016/j.addr.2012.09.010
- 2. Ahearne M, Ahearne M. Introduction to cell – hydrogel mechanosensing. 2014;
- 3. Oyen ML. Mechanical characterisation of hydrogel materials. Int Mater Rev. 2014;59(1):44–59.
- 4. Ozcelik B. Degradable hydrogel systems for biomedical applications [Internet]. Biosynthetic Polymers for Medical Applications. Elsevier Ltd; 2016. 173–188 p. Available from: http://dx.doi.org/10.1016/B978-1-78242-105-4.00007-9
- 5. Draget KI. Alginates. Handb Hydrocoll Second Ed. 2009;807–28.
- 6. Paques JP. Alginate Nanospheres Prepared by Internal or External Gelation with Nanoparticles [Internet]. Microencapsulation and Microspheres for Food Applications. Elsevier Inc.; 2015. 39–55 p. Available from: http://dx.doi.org/10.1016/B978-0-12-800350-3.00004-2
- 7. Rowley JA, Madlambayan G, Mooney DJ. Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials. 1999;20(1):45–53.
- 8. Wang B, Wan Y, Zheng Y, Lee X, Liu T, Yu Z, et al. Alginate-based composites for environmental applications: a critical review. Crit Rev Environ Sci Technol [Internet]. 2019;49(4):318–56. Available from: https://doi.org/10.1080/10643389.2018.1547621
- 9. Andersen T, Auk-Emblem P, Dornish M. 3D Cell Culture in Alginate Hydrogels. Microarrays. 2015;4(2):133–61.
- 10. Kular JK, Basu S, Sharma RI. The extracellular matrix: Structure, composition, age-related differences, tools for analysis and applications for tissue engineering. J Tissue Eng. 2014;5.
- 11. Dalbayrak, Basak, Sonmez E, Kurt H, Isleten Hosoglu M, Kuçuk I, et al. Characterization of a 3D Neuronal-Culture Using Alginate Hydrogels and Optimize For Neuronal Survival and Axon Growth. Nat Appl Sci J. 2021;3(Special).
- 12. Kandemir B, Caglayan B, Hausott B, Erdogan B, Dag U, Demir O, et al. Pea3 transcription factor promotes neurite outgrowth. Front Mol Neurosci. 2014;7(JUNE):1–11.
- 13. Wang H, Wang R, Thrimawithana T, Little PJ, Xu J, Feng ZP, et al. The Nerve Growth Factor Signaling and Its Potential as Therapeutic Target for Glaucoma. Biomed Res Int. 2014;2014.
- 14. Bradshaw RA, Mobley W, Rush RA. Nerve growth factor and related substances: A brief history and an introduction to the international NGF meeting series. Int J Mol Sci. 2017;18(6).