Cell laden alginate-keratin based composite microcapsules containing bioactive glass for tissue engineering applications


Microcapsules based on alginate-keratin, alginate dialdehyde (ADA)-keratin and ADA-keratin-45S5 bioactive glass (BG) were successfully prepared. The samples were characterized by light microscopy, scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The results showed that ADA-based materials possess higher degradation rate compared to alginate–based materials. The incorporation of BG particles (mean particle size: 2.0 µm) improved the bioactivity of the materials. Moreover, the biological properties of the samples were evaluated by encapsulating MG-63 osteosarcoma cells into the microcapsules. The cell viability in all samples increased during 21 days of cultivation. However, the presence of 0.5% BG particle seemed to have initial negative effect on cell growth compared to other samples without BG. On the other hand, the positive effect of CaP formation was visible after 3 weeks in the BG containing samples. The results are relevant to consider the development of cell laden bioinks incorporating inorganic bioactive particles for biofabrication approaches.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. 1.

    Mikos AG, et al. Engineering complex tissues. Tiss Eng. 2006;12:3307–39.

    CAS  Article  Google Scholar 

  2. 2.

    Cross LM, Shah K, Palani S, Peak CW, Gaharwar AK. Gradient nanocomposite hydrogels for interface tissue engineering. Nanomed: Nanotechnol, Biol Med. 2018;14:2465–74.

    CAS  Article  Google Scholar 

  3. 3.

    Yang J, Zhang YS, Yue K, Khademhosseini A. Cell-laden hydrogels for osteochondral and cartilage tissue engineering. Acta Biomater. 2017;57:1–25.

    CAS  Article  Google Scholar 

  4. 4.

    Van Vlierberghe S, Dubruel P, Schacht E. Biopolymer-based hydrogels as scaffolds for tissue engineering applications: a review. Biomacromolecules. 2011;12:1387–408.

    Article  Google Scholar 

  5. 5.

    Nicodemus GD, Bryant SJ. Cell encapsulation in biodegradable hydrogels for tissue engineering applications. Tissue Eng, Part B. 2008;14:149–65.

    CAS  Article  Google Scholar 

  6. 6.

    Gryshkov O, Pogozhykh D, Hofmann N, Pogozhykh O, Mueller T, Glasmacher B. Encapsulating non-human primate multipotent stromal cells in alginate via high voltage for cell-based therapies and cryopreservation. PLoS One. 2014;9:e107911.

    Article  Google Scholar 

  7. 7.

    Zeng Q, Han Y, Li H, Chang J. Bioglass/alginate composite hydrogel beads as cell carriers for bone regeneration. J Biomed Mater Res, Part B. 2014;102B:42–51.

    CAS  Article  Google Scholar 

  8. 8.

    Kim WS, Mooney DJ, Arany PR, Lee K, Huebsch N, Kim J. Adipose tissue engineering using injectable, oxidized alginate hydrogels. Tissue Eng Part A. 2012;18:737–43.

    CAS  Article  Google Scholar 

  9. 9.

    Reakasame S, Boccaccini AR. Oxidized alginate-based hydrogels for tissue engineering applications: a review. Biomacromolecules. 2018;19:3–21.

    CAS  Article  Google Scholar 

  10. 10.

    Moshaverinia A, Chen C, Akiyama K, Ansari S, Xu X, Chee WW, Schricker SR, Shi S. Alginate hydrogel as a promising scaffold for dental-derived stem cells: an in vitro study. J Mater Sci Mater Med. 2012;23:3041–51.

    CAS  Article  Google Scholar 

  11. 11.

    Sarker B, Papageorgiou DG, Silva R, Zehnder T, Gul-E-Noor F, Bertmer M, Kaschta J, Chrissafis K, Detscha R, Boccaccini AR. Fabrication of alginate–gelatin crosslinked hydrogel microcapsules and evaluation of the microstructure and physico-chemical properties. J Mater Chem B. 2014;2:1470–82.

    CAS  Article  Google Scholar 

  12. 12.

    Baniasadi M, Minary-Jolandan M. Alginate-collagen fibril composite hydrogel. Materials. 2015;8:799–814.

    CAS  Article  Google Scholar 

  13. 13.

    Deepthi S, Jayakumar R. Alginate nanobeads interspersed fibrin network as in situ forming hydrogel for soft tissue engineering. Bioact Mater. 2018;3:194–200.

    CAS  Article  Google Scholar 

  14. 14.

    Silva R, Singh R, Sarker B, Papageorgiou DG, Juhasz JA, Roether JA, Cicha I, Kaschta J, Schubert DW, Chrissafis K, Detsch R, Boccaccini AR. Hybrid hydrogels based on keratin and alginate for tissue engineering. J Mater Chem B. 2014;2:5441–51.

    CAS  Article  Google Scholar 

  15. 15.

    Wang HJ, Di L, Ren QS, Wang JY. Applications and degradation of proteins used as tissue engineering materials. Materials. 2009;2:613–35.

    CAS  Article  Google Scholar 

  16. 16.

    Gupta P, Nayak KK. Optimization of keratin/alginate scaffold using RSM and its characterization for tissue engineering. Int J Biol Macromol. 2016;85:141–9.

    CAS  Article  Google Scholar 

  17. 17.

    Srisuwan Y, Srihanam P. Preparation and characterization of keratin/alginate blend microparticles. Adv Mater Sci Eng. 2018;2018: Article ID 8129218.

  18. 18.

    Sarker B, Singh R, Silva R, Roether JA, Kaschta J, Detsch R, Schubert DW, Cicha I, Boccaccini AR. Evaluation of fibroblasts adhesion and proliferation on alginate-gelatin crosslinked hydrogel. PLoS One. 2014;9:e107952.

    Article  Google Scholar 

  19. 19.

    Balakrishnan B, Joshi N, Jayakrishnan A, Banerjee R. Self-crosslinked oxidized alginate/gelatin hydrogel as injectable, adhesive biomimetic scaffolds for cartilage regeneration. Acta Biomater. 2014;10:3650–63.

    CAS  Article  Google Scholar 

  20. 20.

    Sakai S, Yamaguchi S, Takei T, Kawakami K. Oxidized alginate-cross-linked alginate/gelatin hydrogel fibers for fabricating tubular constructs with layered smooth muscle cells and endothelial cells in collagen gels. Biomacromolecules. 2008;9:2036–41.

    CAS  Article  Google Scholar 

  21. 21.

    Hench L. Bioceramics. J Am Ceram Soc. 1998;81:1705–28.

    CAS  Article  Google Scholar 

  22. 22.

    Leite ÁJ, Sarker B, Zehnder T, Silva R, Mano JF, Boccaccini AR. Bioplotting of a bioactive alginate dialdehyde-gelatin composite hydrogel containing bioactive glass nanoparticles. Biofabrication. 2016;8:035005.

    Article  Google Scholar 

  23. 23.

    Hench LL. Some comments on bioglass: four eras of discovery and development. Biomed Glass. 2015;1:1–11.

    Google Scholar 

  24. 24.

    Midha S, Kumar S, Sharma A, Kaur K, Shi X, Naruphontjirakul P, Jones JR, Ghosh S. Silk fibroin-bioactive glass based advanced biomaterials: towards patient-specific bone grafts. Biomed Mater. 2018;13:055012.

    Article  Google Scholar 

  25. 25.

    Rottensteiner-Brandl U, Detsch R, Sarker B, Lingens L, Köhn K, Kneser U, Bosserhoff AK, Horch RE, Boccaccini AR, Arkudas A. Encapsulation of rat bone marrow derived mesenchymal stem cells in alginate dialdehyde/gelatin microbeads with and without nanoscaled bioactive glass for in vivo bone tissue engineering. Materials. 2018;11:1880.

    Article  Google Scholar 

  26. 26.

    Wheeler TS, Sbravati ND, Janorkar AV. Mechanical & cell culture properties of elastin-like polypeptide, collagen, bioglass, and carbon nanosphere composites. Ann Biomed Eng. 2013;41:2042–55.

    Article  Google Scholar 

  27. 27.

    Zehnder T, Boccaccini AR, Detsch R. Biofabrication of a co-culture system in an osteoid-like hydrogel matrix. Biofabrication. 2017;9:025016.

    Article  Google Scholar 

  28. 28.

    Peterson GL. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977;83:346–56.

    CAS  Article  Google Scholar 

  29. 29.

    Laemmli UK. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature. 1970;227:680–5.

    CAS  Article  Google Scholar 

  30. 30.

    Kokubo T, Takadama H. How useful is SBF in predicting in vivo bone bioactivity? Biomaterials. 2006;27:2907–15.

    CAS  Article  Google Scholar 

  31. 31.

    Guo Z, Yang C, Zhou Z, Chen S, Li F. Characterization of biodegradable poly(lactic acid) porous scaffolds prepared using selective enzymatic degradation for tissue engineering. RSC Adv. 2017;7:34063.

    CAS  Article  Google Scholar 

  32. 32.

    Sarker B, Li W, Zheng K, Detsch R, Boccaccini AR. Designing porous bone tissue engineering scaffolds with enhanced mechanical properties from composite hydrogels composed of modified alginate, gelatin, and bioactive glass. ACS Biomater Sci Eng. 2016;2:2240–54.

    CAS  Article  Google Scholar 

  33. 33.

    Vasconcelos A, Freddi G, Cavaco-Paulo A. Biodegradable materials based on silk fibroin and keratin. Biomacromolecules. 2008;9:1299–305.

    CAS  Article  Google Scholar 

  34. 34.

    Zheng K, Solodovnyk A, Li W, Goudouri OM, Stähli C, Nazhat SN, Boccaccini AR. Aging time and temperature effects on the structure and bioactivity of gel-derived 45S5 glass-ceramics. J Am Ceram Soc. 2015;98.1:30–38.

    Article  Google Scholar 

  35. 35.

    Reznikov N, Steele JAM, Fratzl P, Stevens MM. Materials science vision of extracellular matrix mineralization. Nat Rev Mater. 2016;1:16041.

    CAS  Article  Google Scholar 

  36. 36.

    Kristiansen KA, Tomren HB, Christensen BE. Periodate oxidized alginates: depolymerization kinetics. Carbohydr Polym. 2011;86:1595–601.

    CAS  Article  Google Scholar 

  37. 37.

    Rottensteiner U, Sarker B, Heusinger D, Dafinova D, Rath SN, Beier JP, Kneser U, Horch RE, Detsch R, Boccaccini AR, Arkudas A. In vitro and in vivo biocompatibility of alginate dialdehyde/gelatin hydrogels with and without nanoscaled bioactive glass for bone tissue engineering applications. Materials. 2014;7:1957–74.

    CAS  Article  Google Scholar 

Download references


Supachai Reakasame acknowledges the German Academic Exchange Service (DAAD) for financial support. We thank Dr.-Ing. Kai Zheng and Dr. Aldo Leal-Egaña (Institute of Biomaterials, University of Erlangen-Nuremberg) for helpful discussions.

Author information



Corresponding author

Correspondence to Aldo R. Boccaccini.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Reakasame, S., Trapani, D., Detsch, R. et al. Cell laden alginate-keratin based composite microcapsules containing bioactive glass for tissue engineering applications. J Mater Sci: Mater Med 29, 185 (2018). https://doi.org/10.1007/s10856-018-6195-5

Download citation