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Bioprinted PDLSCs with high-concentration GelMA hydrogels exhibit enhanced osteogenic differentiation in vitro and promote bone regeneration in vivo

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Abstract

Objectives

We aimed to explore the osteogenic potential of periodontal ligament stem cells (PDLSCs) in bioprinted methacrylate gelatine (GelMA) hydrogels in vitro and in vivo.

Materials and methods

PDLSCs in GelMA hydrogels at various concentrations (3%, 5%, and 10%) were bioprinted. The mechanical properties (stiffness, nanostructure, swelling, and degradation properties) of bioprinted constructs and the biological properties (cell viability, proliferation, spreading, osteogenic differentiation, and cell survival in vivo) of PDLSCs in bioprinted constructs were evaluated. Then, the effect of bioprinted constructs on bone regeneration was investigated using a mouse cranial defect model.

Results

Ten percent GelMA printed constructs had a higher compression modulus, smaller porosity, lower swelling rate, and lower degradation rate than 3% GelMA. PDLSCs in bioprinted 10% GelMA bioprinted constructs showed lower cell viability, less cell spreading, upregulated osteogenic differentiation in vitro, and lower cell survival in vivo. Moreover, upregulated expression of ephrinB2 and EphB4 protein and their phosphorylated forms were found in PDLSCs in 10% GelMA bioprinted constructs, and inhibition of eprhinB2/EphB4 signalling reversed the enhanced osteogenic differentiation of PDLSCs in 10% GelMA. The in vivo experiment showed that 10% GelMA bioprinted constructs with PDLSCs contributed to more new bone formation than 10% GelMA constructs without PDLSCs and constructs with lower GelMA concentrations.

Conclusions

Bioprinted PDLSCs with high-concentrated GelMA hydrogels exhibited enhanced osteogenic differentiation partially through upregulated ephrinB2/EphB4 signalling in vitro and promoted bone regeneration in vivo, which might be more appropriate for future bone regeneration applications.

Clinical relevance

Bone defects are a common clinical oral problem. Our results provide a promising strategy for bone regeneration through bioprinting PDLSCs in GelMA hydrogels.

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Data availability

All data generated or analyzed during this study are included in this published article.

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Acknowledgements

The authors thank the Public Experimental Center of Xuzhou Medical University for their help and support.

Funding

This study was supported by National Nature Science Foundation of China (grant no. 82201071), Xuzhou Key Research and Development Project (grant no. KC21236), Research Program of the Jiangsu Provincial Health Commission (grant no. M2020091), and Innovation and Entrepreneurship Training Program for College Students of Jiangsu Province (202110313009Z).

Author information

Authors and Affiliations

  1. School of Stomatology, Xuzhou Medical University, No. 209 Tongshan Road, Xuzhou, 221004, Jiangsu, China

    Yaru Zhu, Wen Wang, Qiyu Chen, Tianshui Ren, Yanbin Qi, Changyong Yuan & Penglai Wang

  2. Affiliated Stomatological Hospital of Xuzhou Medical University, No.130 Huaihai West Road, Xuzhou, 221000, Jiangsu, China

    Wen Wang, Jianguang Yang, Gan Li, Changyong Yuan & Penglai Wang

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  1. Yaru Zhu
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  2. Wen Wang
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  9. Penglai Wang
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Contributions

Y.Z. and W.W. contributed to conceptualization (lead); writing—original draft (lead); formal analysis (lead); writing— review and editing (equal). Q.C. and T.R. assisted with methodology (lead); writing—review and editing (equal). J.Y. and G.L. and Y.Q. provided software (lead); writing—review and editing (equal). C.Y. contributed to conceptualization (supporting); writing—review and editing (lead). P.W. assisted with conceptualization (supporting); writing—original draft (supporting); writing—review and editing (equal).

Corresponding authors

Correspondence to Changyong Yuan or Penglai Wang.

Ethics declarations

Ethical approval

Human third molars were obtained from healthy volunteers with their written informed consent. Animal experiments in this study were approved by the Experimental Animal Ethics Committee of Xuzhou Medical University, approval No.202208S107. All experimental procedures were performed in accordance with the corresponding guidelines and regulations. All animals were kept in a pathogen-free environment and there were no incidents of animal cruelty.

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The authors declare no competing interests.

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Yaru Zhu and Wen Wang contributed equally to this work and share first authorship.

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Zhu, Y., Wang, W., Chen, Q. et al. Bioprinted PDLSCs with high-concentration GelMA hydrogels exhibit enhanced osteogenic differentiation in vitro and promote bone regeneration in vivo. Clin Oral Invest 27, 5153–5170 (2023). https://doi.org/10.1007/s00784-023-05135-7

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