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Effect of 3-dimensional Collagen Fibrous Scaffolds with Different Pore Sizes on Pulp Regeneration

  • Qianli Zhang
    Affiliations
    Department of Cariology and Endodontology, National Engineering Laboratory for Digital and Material Technology, Peking Key Laboratory of Digital Stomatology, Peking University School and Hospital of Stomatology, Beijing, China

    4th Dental Department, Peking University School and Hospital of Stomatology, Beijing, China
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  • Chongyang Yuan
    Affiliations
    Department of Cariology and Endodontology, National Engineering Laboratory for Digital and Material Technology, Peking Key Laboratory of Digital Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
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  • Li Liu
    Affiliations
    Beijing Engineering Research Centre of Advanced Elastomers, Beijing University of Chemical Technology, Beijing, China
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  • Shipeng Wen
    Correspondence
    Address requests for reprints to Dr Shipeng Wen, Beijing Engineering Research Centre of Advanced Elastomers, Beijing University of Chemical Technology, Beijing 100029, China.
    Affiliations
    Beijing Engineering Research Centre of Advanced Elastomers, Beijing University of Chemical Technology, Beijing, China
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  • Xiaoyan Wang
    Correspondence
    Dr Xiaoyan Wang, Department of Cariology and Endodontology, National Engineering Laboratory for Digital and Material Technology, Peking Key Laboratory of Digital Stomatology, Peking University School and Hospital of Stomatology, Beijing 100081, China.
    Affiliations
    Department of Cariology and Endodontology, National Engineering Laboratory for Digital and Material Technology, Peking Key Laboratory of Digital Stomatology, Peking University School and Hospital of Stomatology, Beijing, China
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Published:October 18, 2022DOI:https://doi.org/10.1016/j.joen.2022.10.007

      Abstract

      Introduction

      In this study, we generated a 3-dimensional (3D) collagen fibrous scaffold for potential pulp regeneration and investigated the influence of various pore sizes of these scaffolds on proliferation, odontoblastic differentiation of human dental pulp cells (hDPCs), and subsequent tissue formation during pulp regeneration.

      Methods

      Electrospinning followed by freeze-drying was used to fabricate 3D fibrous collagen scaffolds. hDPCs were cultured on these scaffolds. Cell growth was detected by a Cell Counting Kit-8 assay and observed via scanning electron microscopy. Odontogenic genes and protein expression were analyzed by real-time reverse transcription polymerase chain reaction and immunofluorescence staining. The formation of mineralized nodules was tested by von Kossa staining, scanning electron microscopy, and energy-dispersive X-ray microanalysis. Subcutaneous transplantation of the seeded scaffold/tooth fragments into nude mice was performed to observe tissue formation for pulp regeneration.

      Results

      Collagen 3D fibrous scaffolds with 3 distinct mean pore sizes (approximately 20 μm, 65 μm, and 145 μm) were fabricated, which showed good biocompatibility and bioactivity. Scaffolds with larger mean pore sizes of 65 and 145 μm improved hDPC ingrowth and proliferation, with the 65-μm scaffold group presenting the highest level of odontogenic gene expression (DSPP and DMP-1), protein expression (DMP-1), mineralized area ratio, and vascular pulplike tissue formation after 6 weeks of subcutaneous implantation.

      Conclusions

      The pore size of collagen 3D fibrous scaffolds significantly affected cell adhesion, proliferation, odontoblastic differentiation, and tissue rehabilitation. Scaffolds with a mean pore size of 65 μm presented superior results and could be an alternative for pulp regeneration.

      Key Words

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