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The effect of different cross-linking conditions of EDC/NHS on type II collagen scaffolds: an in vitro evaluation

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Abstract

The purpose of this paper is to analyze the properties of porcine cartilage type II collagen scaffolds crosslinked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide/N-hydroxy-succinamide (EDC/NHS) under different conditions. The porous EDC/NHS-crosslinked scaffolds were obtained through a two-step freeze-drying process. To determine the optimal crosslinking condition, we used different solvents and various crosslinking temperatures to prepare the scaffolds. Three crosslinking solutions were prepared with different solvents, photographs were taken with a flash in the darkroom, and light transmission was observed. Type II collagen was crosslinked on a horizontal shaker at a speed of 60 r/min according to the above grouping conditions, and then the structural change of the scaffold in each group was observed. To investigate the swelling ratio and the in vitro degradation of the collagen scaffold, tests were also carried out by immersion of the scaffolds in a PBS solution and digestion in type II collagenase, respectively. The influence of the scaffolds on the proliferation of chondrocytes was assessed by the methyl thiazolyl tetrazolium colorimetric assay. The morphology of the crosslinked scaffolds cocultured with chondrocytes was characterized by a scanning electron microscope. The results proved that 75% alcohol and a crosslinking temperature of 37 °C are recommended. Collagen fibrils are more densely packed after crosslinking with EDC/NHS and have a more uniform structure than that of noncrosslinked ones. The EDC-crosslinked scaffolds possessed excellent mechanical property and biocompatibility.

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Acknowledgements

This study was supported by the Project of invigorating Health Care through Science, Technology and Education (Jiangsu Provincial Medical Youth Talent), Changzhou High-level Medical Talents Training Project (2016CZBJ029).

Funding

This study was funded by the Project of invigorating Health Care through Science, Technology and Education (Jiangsu Provincial Medical Youth Talent), Changzhou High-level Medical Talents Training Project (2016CZBJ029).

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Author notes

  1. Lu-Ming Nong, Dong Zhou and Dong Zheng have contributed equally as first authors.

Authors and Affiliations

  1. Orthopedics Department, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, 213000, Jiangsu Province, China

    Lu-Ming Nong, Dong Zhou, Dong Zheng, Yu-Qing Jiang, Nan-Wei Xu, Gong-Yin Zhao, Hui Wei & Long Han

  2. Laboratory Department, The Affiliated Changzhou No.2 People’s Hospital of Nanjing Medical University, Changzhou, 213000, Jiangsu Province, China

    Si-Yuan Zhou

  3. General Surgery Department, Huaiyin Hospital of Huai’an City, Huai’an, 223300, Jiangsu Province, China

    Hui Han

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  1. Lu-Ming Nong
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Correspondence to Long Han.

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Nong, LM., Zhou, D., Zheng, D. et al. The effect of different cross-linking conditions of EDC/NHS on type II collagen scaffolds: an in vitro evaluation. Cell Tissue Bank 20, 557–568 (2019). https://doi.org/10.1007/s10561-019-09790-7

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