Skip to main content

Advertisement

Log in

Fabrication and preliminary in vitro evaluation of ultraviolet-crosslinked electrospun fish scale gelatin nanofibrous scaffolds

  • TISSUE ENGINEERING CONSTRUCTS AND CELL SUBSTRATES
  • Original Research
  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

This study aimed to explore a potential use of fish scale-derived gelatin nanofibrous scaffolds (GNS) in tissue engineering due to their biological and economical merits. Extraction of gelatin was achieved via decalcification, sonication and lyophilization of mixed fish scales. To fabricate nano-scale architecture of scaffolds analogous to natural extracellular matrix, gelatin was rendered into nanofibrous matrices through 6-h electrospinning, resulting in the average diameter of 48 ± 12 nm. In order to improve the water-resistant ability while retaining their biocompatibility, GNS were physically crosslinked with ultraviolet (UV) irradiation for 5 min (UGN5), 10 min (UGN10) and 20 min (UGN20). On average, the diameter of nanofibers increased by 3 folds after crosslinking, however, Fourier transform infrared spectroscopy analysis confirmed that no major alterations occurred in the functional groups of gelatin. A degradation assay showed that UGN5 and UGN10 scaffolds remained in minimum essential medium for 14 days, while UGN20 scaffolds degraded completely after 10 days. All UGN scaffolds promoted adhesion and proliferation of human keratinocytes, HaCaT, without causing an apparent cytotoxicity. UGN5 scaffolds were shown to stimulate a better growth of HaCaT cells compared to other scaffolds upon 1 day of incubation, whereas UGN20 had a long-term effect on cells exhibiting 25% higher cell proliferation than positive control after 7 days. In the wound scratch assay, UGN5 scaffolds induced a rapid cell migration closing up to 79% of an artificial wound within 24 h. The current findings provide a new insight of UGN scaffolds to serve as wound dressings in the future.

In the wound scratch assay, UGN5 induced a rapid cell migration closing up to 79% of an artificial wound within 24 h.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Singer AJ, Dagum AB. Current management of acute cutaneous wounds. N Engl J Med. 2008;359:1037–46.

    CAS  Google Scholar 

  2. Demidova-Rice TN, Hamblin MR, Herman IM. Acute and impaired wound healing: pathophysiological and current methods for drug delivery, part 1: normal and chronic wounds: biology, causes, and approaches to care. Adv Skin Wound Care. 2012;25:304–14.

    Google Scholar 

  3. Ogurtsova K, da Rocha Fernandes JD, Huang Y, et al. IDF Diabetes Atlas: Global estimates for the prevalence of diabetes for 2015 and 2040. Diabetes Res Clin Pract. 2017;128:40–50.

    CAS  Google Scholar 

  4. Posnett J, Franks PJ. The burden of chronic wounds in the UK. Nurs Times. 2008;104:44–5.

    Google Scholar 

  5. Sood A, Granick MS, Tomaselli NL. Wound dressings and comparative effectiveness data. Adv Wound Care. 2014;3:511–29.

    Google Scholar 

  6. Frantz C, Stewart KM, Weaver VM. The extracellular matrix at a glance. J Cell Sci. 2010;123(Pt 24):4195–200.

    CAS  Google Scholar 

  7. Gnavi S, Fornasari BE, Tonda-Turo C, et al. The influence of electrospun fibre size on Schwann cell behavior and axonal outgrowth. Mater Sci Eng: C. 2015;48:620–31.

    CAS  Google Scholar 

  8. Ghaly AE, Ramakrishnan VV, Brooks MS, Budge SM, Dave D. Fish processing wastes as a potential source of protein. Amino Acids and Oils: A Critical Review. J Microb Biochem Technol. 2013;5:107–29.

    Google Scholar 

  9. Boran G, Mulvaney SJ, Regenstein JM. Rheological properties of gelatin from silver carp skin compared to commercially available gelatins from different sources. J Food Sci. 2010;75:E565–71.

    CAS  Google Scholar 

  10. Herpandi H, Huda N, Adzitey F. Fish bone and scale as a potential source of Halal gelatin. J Fish Aquat Sci. 2011;6:379–89.

    CAS  Google Scholar 

  11. Badami AS, Kreke MR, Thompson MS, Riffle JS, Goldstein AS. Effect of fiber diameter on spreading, proliferation and differentiation of osteoblastic cells on electrospun poly (lactic acid) substrates. Biomaterials. 2006;24:596–606.

    Google Scholar 

  12. Chua KN, Chai C, Lee PC, et al. Surface-aminated electrospun nanofibers enhance adhesion and expansion of human umbilical cord blood hematopoietic stem/progenitor cells. Biomaterials. 2006;27:6043–51.

    CAS  Google Scholar 

  13. Agarwal S, Wendorff JH, Greiner A. Use of electrospinning technique for biomedical applications. Polymer. 2008;49:5603–21.

    CAS  Google Scholar 

  14. Bigi A, Cojazzi G, Panzavolta S, Rubini K, Roveri N. Mechanical and thermal properties of gelatin films at different degrees of glutaraldehyde crosslinking. Biomaterials. 2001;22:763–8.

    CAS  Google Scholar 

  15. Zhang YZ, Venugopal J, Huang ZM, Lim CT, Ramakrishna S. Crosslinking of the electrospun gelatin nanofibers. Polymer. 2006;47:2911–7.

    CAS  Google Scholar 

  16. Sisson K, Zhang C, Farach-Carson MC, Chase DB, Rabolt JF. Fiber diameters control osteoblastic cell migration and differentiation in electrospun gelatin. J Biomed Mater Res Part A. 2010;94:1312–20.

    Google Scholar 

  17. Huang-Lee LL, Cheung DT, Nimni ME. Biochemical changes and cytoxicity associated with the degradatin of polymeric glutaraldehydge derived crosslinks. J Biomedical Materials Part A. 1990;24:1185–201.

    CAS  Google Scholar 

  18. Gough JE, Scotchford CA, Downes S. Cytotoxicity of glutaraldehyde crosslinked collagen/poly (vinyl alcohol) films is by the mechanism of apoptosis. J Biomed Mater Res A. 2002;61:121–30.

    CAS  Google Scholar 

  19. Weadock KS, Miller EJ, Bellincampi LD, Zawadsky JP, Dunn MG. Physical crosslinking of collagen fibers: Comparison of ultraviolet irradiation and dehydrothermal treatment. J of Biomed Mater Res. 1995;29:1373–9.

    CAS  Google Scholar 

  20. Kato Y, Nishikawa T, Kawakishi S. Formation of protein‐bound 3,4‐dihydroxyphenylalanine in collagen types i and iv exposed to ultraviolet light. Photochem Photobiol. 1995;61:367–72.

    CAS  Google Scholar 

  21. Lew DH, Liu PHT, Orgill DP. Optimization of UV cross‐linking density for durable and nontoxic collagen GAG dermal substitute. J Biomed Mater Res Part B: Appl Biomater. 2007;82B:51–6.

    CAS  Google Scholar 

  22. Lin WH, Tsai WB. In situ UV-crosslinking gelatin electrospun fibers for tissue engineering applications. Biofabrication. 2013;5:035008.

    Google Scholar 

  23. Masutani EM, Kinoshita CK, Tanaka TT, Ellison AK, Yoza BA. Increasing thermal stability of gelatin by UV-induced cross-linking with glucose. Int J Biomate. 2014;2014:979636.

    Google Scholar 

  24. Mori M, Hamamoto A, Nakano M. Effects of ultraviolet LED on bacteria. World Congress on Medical Physics and Biomedical Engineering. 2006;2:1327–30.

    Google Scholar 

  25. Tu ZC, Huang T, Wang H. Physico-chemical properties of gelatin from bighead carp (Hypophthalmichthys nobilis) scales by ultrasound-assisted extraction. J Food Sci Technol. 2015;52:2166–74.

    CAS  Google Scholar 

  26. Huang C-Y, Kuo J-M, Wu S-J, Tsai H-T. Isolation and characterization of fish scale collagen from tilapia (Oreochromis sp.) by a novel extrusion-hydro-extraction process. Food Chem. 2016;190:997–1006.

    CAS  Google Scholar 

  27. Gómez-Guillén MC, Turnay J, Fernández-Dı́az MD, Ulmo N, Lizarbe MA, Montero P. Structural and physical properties of gelatin extracted from different marine species: a comparative study. Food Hydrocoll. 2002;16:25–34.

    Google Scholar 

  28. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680–5.

    CAS  Google Scholar 

  29. Nuge T, Tshai KY, Lim SS, Nordin N, Hoque ME. Preparation and characterization of CU-, FE-, AG-, ZN- and NI- doped gelatin nanofibers for possible applications in antibacterial nanomedicine. J Eng Sci Technol. 2017;12:68–81.

    Google Scholar 

  30. Zhou P, Mulvaney SJ, Regenstein JM. Properties of Alaska pollock skin gelatin: A comparison with tilapia and pork skin gelatins. J Food Sci. 2006;71:C313–21.

    CAS  Google Scholar 

  31. Lin CC, Chiou TK, Sung WC. Characteristics of Gelatin from Giant Grouper (Epinephelus Lanceolatus) skin. Int J Food Prop. 2015;18:2339–48.

    CAS  Google Scholar 

  32. Song X, Dong P, Gravesande J, Cheng B, Xing J. UV-mediated solid-state cross-linking of electrospinning nanofibers of modified collagen. Int J Biol Macromol. 2018;120:2086–93.

    CAS  Google Scholar 

  33. Yslas EI, Cavallo P, Acevedo DF, Barbero CA, Rivarola VA. Cysteine modified polyaniline films improve biocompatibility for two cell lines. Mater Sci Eng: C. 2015;51:51–6.

    CAS  Google Scholar 

  34. Lutolf MP, Hubbell JA. Synthetic biomaterials as instructive extracellular microenvironments for morphogenesis in tissue engineering. Nat Biotechnol. 2005;23:47–55.

    CAS  Google Scholar 

  35. Xu CY, Inai R, Kotaki M, Ramakrishna S. Aligned biodegradable nanofibrous structure: a potential scaffold for blood vessel engineering. Biomaterials. 2004;25:877–86.

    CAS  Google Scholar 

  36. Kumbar SG, Nukavarapu SP, James R, Nair LS, Laurencin CT. Electrospun poly (lactic acid-co-glycolic acid) scaffolds for skin tissue engineering. Biomaterials. 2008;29:4100–7.

    CAS  Google Scholar 

  37. Min BM, Lee G, Kim SH, Nam YS, Lee TS, Park WH. Electrospinning of silk fibroin nanofibers and its effect on the adhesion and spreading of normal human keratinocytes and fibroblasts in vitro. Biomaterials. 2004;25:1289–97.

    CAS  Google Scholar 

  38. Cooper DR, Davidson RJ. The effect of ultraviolet irradiation on collagen-fold formation. Biochemical J. 1966;98:655–61.

    CAS  Google Scholar 

  39. Fujimori E. Changes induced by ozone and ultraviolet light in type I collagen. European J Biochem. 1985;152:299–306.

    CAS  Google Scholar 

  40. Sionkowska A. Modification of collagen films by ultraviolet irradiation. Polym Degrad Stab. 2000;68:147–51.

    CAS  Google Scholar 

  41. Wang X, Li X, Yost MJ. Microtensile testing of collagen fibril for cardiovascular tissue engineering. J Biomed Mater Res. 2005;74:263–8.

    Google Scholar 

  42. Muyonga JH, Cole CGB, Duodu KG. Fourier transform infrared (FTIR) spectroscopic study of acid soluble collagen and gelatin from skins and bones of young and adult Nile perch (Lates niloticus). Food Chem. 2004;86:325–32.

    CAS  Google Scholar 

  43. Byler DM, Susi H. Examination of the secondary structure of proteins by deconvolved FTIR spectra. Biopolymers. 1986;25:469–87.

    CAS  Google Scholar 

  44. Ohan MP, Weadock KS, Dunn MG. Synergistic effects of glucose and ultraviolet irradiation on the physical properties of collagen. J Biomed Mater Res. 2002;60:384–91.

    CAS  Google Scholar 

  45. Davidenko N, Bax DV, Schuster CF, Farndale RW, Hamaia SW, Best SM, Cameron RE. Optimisation of UV irradiation as a binding site conserving method for crosslinking collagen-based scaffolds. J Mater Sci: Mater Med. 2016;27:14.

    Google Scholar 

  46. Cooper DR, Davidson RJ. Tthe effect of ultraviolet irradiation on soluble collagen. Biochem J. 1965;97:139–47.

    CAS  Google Scholar 

  47. Sudbeck BD, Pilcher BK, Welgus HG, Parks WC. Induction and repression of collagenase-1 by keratinocytes is controlled by distinct components of different extracellular matrix compartments. J Biol Chem. 1997;272:22103–10.

    CAS  Google Scholar 

  48. Rahsaz M, Geramizadeh B, Kaviani M, Marzban S. Gelatin for purification and proliferation of primary keratinocyte culture for use in chronic wounds and burns. Exp Clin Transplant. 2015;13:361–5.

    Google Scholar 

  49. Liang HC, Chang WH, Liang HF, Lee MH, Sung HW. Crosslinking structures of gelatin hydrogels crosslinked with genipin or a water‐soluble carbodiimide. J Appl Polym Sci. 2004;91:4017–26.

    CAS  Google Scholar 

  50. Kanade S, Nataraj G, Ubale M, Mehta P. Fluorescein diacetate vital staining for detecting viability of acid-fast bacilli in patients on antituberculosis treatment. Int J Mycobacteriol. 2016;5:294–98.

    Google Scholar 

  51. Fu X, Xu M, Liu J, Qi Y, Li S, Wang H. Regulation of migratory activity of human keratinocytes by topography of multiscale collagen-containing nanofibrous matrices. Biomaterials. 2014;35:1496–506.

    CAS  Google Scholar 

  52. Jafari J, Emami SH, Samadikuchaksaraei A, Bahar MA, Gorjipour F. Electrospun chitosan–gelatin nanofiberous scaffold: Fabrication and in vitro evaluation. Biomed Mater Eng. 2011;21:99–112.

    CAS  Google Scholar 

  53. Das RP, Jain AK, Ramesh V. Current concepts in the pathogenesis of psoriasis. Indian J Dermatol. 2009;54:7–12.

    Google Scholar 

  54. Takashi K, Folmer J, Coulombe PA. Increased expression of keratin 16 causes anomalies in cytoarchitecture and keratinization in transgenic mouse skin. J Cell Biol. 1994;127:505–20.

    Google Scholar 

Download references

Acknowledgements

This study was conducted under the collaboration of Faculty of Science and Faculty of Engineering and funded by The University of Nottingham Malaysia Campus.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Siew Shee Lim or Hwei-San Loh.

Ethics declarations

Conflict of interest

The authors declare that they do not have any conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Beishenaliev, A., Lim, S.S., Tshai, K.Y. et al. Fabrication and preliminary in vitro evaluation of ultraviolet-crosslinked electrospun fish scale gelatin nanofibrous scaffolds. J Mater Sci: Mater Med 30, 62 (2019). https://doi.org/10.1007/s10856-019-6264-4

Download citation

  • Published:

  • DOI: https://doi.org/10.1007/s10856-019-6264-4

Navigation