Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (22): 3478-3483.doi: 10.12307/2024.534

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Effect of dura mater on enhancement of cranial osteogenesis in rats

An Ran1, 2, 3, 4, Shao Guo5, Zhang Chunyang2, 3, 4   

  1. 1Baotou  Medical  College  of Inner  Mongolia  University  of Science  and  Technology, Baotou 014000, Inner Mongolia Autonomous Region, China; 2Department of Neurosurgery, First Affiliated Hospital of Baotou Medical College of Inner Mongolia University of Science and Technology, Baotou 014010, Inner Mongolia Autonomous Region, China; 3Institute of Neurosurgical Diseases (Translational Medicine), Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China; 4Inner Mongolia Autonomous Region Bone Tissue Regeneration and Damage Repair Engineering Technology Center, Baotou 014010, Inner Mongolia Autonomous Region, China; 5Translational Medicine Center of The Third People’s Hospital of Longgang District of Shenzhen City, Shenzhen 518100, Guangdong Province, China
  • Received:2023-01-07 Accepted:2023-11-16 Online:2024-08-08 Published:2024-01-20
  • Contact: Zhang Chunyang, Professor, Chief physician, Department of Neurosurgery, First Affiliated Hospital of Baotou Medical College of Inner Mongolia University of Science and Technology, Baotou 014010, Inner Mongolia Autonomous Region, China; Institute of Neurosurgical Diseases (Translational Medicine), Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China; Inner Mongolia Autonomous Region Bone Tissue Regeneration and Damage Repair Engineering Technology Center, Baotou 014010, Inner Mongolia Autonomous Region, China
  • About author:An Ran, Master candidate, Baotou Medical College of Inner Mongolia University of Science and Technology, Baotou 014000, Inner Mongolia Autonomous Region, China; Department of Neurosurgery, First Affiliated Hospital of Baotou Medical College of Inner Mongolia University of Science and Technology, Baotou 014010, Inner Mongolia Autonomous Region, China; Institute of Neurosurgical Diseases (Translational Medicine), Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China; Inner Mongolia Autonomous Region Bone Tissue Regeneration and Damage Repair Engineering Technology Center, Baotou 014010, Inner Mongolia Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China, No. 82160250, 81960238 (to ZCY); Funding for the National Clinical Key Specialty Construction Project (to ZCY); Graduate Student Research and Innovation Program of Baotou Medical College, No. bycx2023013 (to AR)

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Abstract: BACKGROUND: The dura mater and skull are physically and functionally related, although there have been few investigations on primary extraction of dura mater and cranial cells, as well as co-culture of the two. The use of primary cells to investigate the influence of the dura mater on the skull is novel, and it is hoped that it may give a theoretical foundation for therapeutic therapy.
OBJECTIVE: Rat dura mater and cranial bone cells were retrieved in situ to observe the influence of dura mater on cranial bone proliferation and differentiation, as well as to get a basic knowledge of the involvement of Twist1 in this process.
METHODS: The enzyme digestion method was used in conjunction with the tissue block method to extract dural cells and cranial osteoblasts from rats within three days of birth. Immunofluorescence staining was used to identify the extracted cells, and alizarin red staining was used to identify and evaluate cranial osteoblasts and their mineralization ability. After co-culturing dural cells and cranial osteoblasts, real-time PCR was utilized to identify the expression of genes associated to cranial osteoblast proliferation and osteogenesis, as well as Twist1.
RESULTS AND CONCLUSION: (1) Morphology: The retrieved dural cells had morphological traits similar to fibroblasts, while the osteoblasts were spindle-shaped. (2) Cell identification: immunofluorescence staining revealed that extracted dural cells expressed high levels of vimentin and cranial osteoblasts expressed high levels of alkaline phosphatase; cranial osteoblasts were stained with alizarin red 28 days after osteogenic induction, and obvious mineralized nodules were observed. (3) Real time PCR detection showed that the co-culture group had higher levels of PCNA, alkaline phosphatase, and RUNX2 mRNA expression than the control group (P < 0.01); however, Twist1 mRNA expression was lower (P < 0.01). (4) The findings showed that the primary extracted cranial osteoblasts had a high mineralization capacity, and that the dura mater was a key factor in promoting cranial growth and development and osteogenic differentiation, with Twist1 playing a key role in this process.       

Key words: primary cell, dura mater, skull, osteogenesis, Twist1, osteogenic differentiation

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