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Enzymatic modification of eggshell membrane with the application in biomimetic scaffold

  • Materials for life sciences
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Abstract

Biomimetic scaffolds that induce bone growth in defects have always been the main focus of bone regeneration research. This study was to develop biomineralization scaffolds using eggshell membranes as the bio-template. The structural and mineralizing characteristics of the inner and outer eggshell membrane (IM and OM) before and after alkaline protease and sodium sulfite treatments were investigated. Electron microscopy showed that both the IM and OM had a core-mantel structure. IM had a thinner mantel layer and more uniform fiber network structure than OM, exhibiting small average pore size and high specific surface area. The mineralization of IM and OM only occurred on the fibers surface. However, enzymatic treatment caused partial destruction of the mantel layer, which allowed the formation of mineral crystals. Specifically, the IM and enzyme-treated IM (EIM) fibers had better mineralization potentials than OM and the enzyme-treated OM (EOM), respectively. Hydroxyapatite-accumulated EIM (EIMHA) showed stronger hydrophilicity, higher BSA fixation ability, and simulated tissue fluid absorption ability. Moreover, the EIMHA facilitated the cell adhesion of the MC3T3-E1 cells and showed a strong cell proliferation activity. In summary, eggshell membranes treated with alkaline protease and sodium sulfite has a high potential to be used as a biomimetic mineralization scaffold.

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Abbreviations

ESM:

Eggshell membrane

IM:

Inner eggshell membrane

OM:

Outer eggshell membrane

LM:

Limiting membrane

OMHA:

Mineralized outer eggshell membrane

IMHA:

Mineralized inner eggshell membrane

EOMHA:

Enzyme-hydrolyzed OM/ hydroxyapatite

EIMHA:

Enzyme-hydrolyzed IM/hydroxyapatite

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Acknowledgements

We acknowledge the financial support from National Natural Science Foundation of China (32072237, 32072158).

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Author notes

  1. Xing Fu, Qian-Cheng Zhao contribute equally to this paper and should be considered as co-first author

Authors and Affiliations

  1. College of Food Science and Technology, National Research and Development Centre for Egg Processing, Huazhong Agricultural University, Hubei, 430070, Wuhan, People’s Republic of China

    Xing Fu, Qian-cheng Zhao, Mei-hu Ma & Xi Huang

  2. Department of Animal Science, Iowa State University, Ames, IA, 50011, USA

    Dong Uk Ahn

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  1. Xing Fu
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Correspondence to Xi Huang.

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Fu, X., Zhao, Qc., Ahn, D.U. et al. Enzymatic modification of eggshell membrane with the application in biomimetic scaffold. J Mater Sci 57, 14993–15007 (2022). https://doi.org/10.1007/s10853-022-07557-2

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