中国组织工程研究2024，Vol.28Issue(22)：3478-3483,6.DOI:10.12307/2024.534
大鼠硬脑膜对颅骨成骨增强的影响
Effect of dura mater on enhancement of cranial osteogenesis in rats
摘要
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.关键词
原代细胞/硬脑膜/颅骨/成骨/Twist1/成骨分化Key words
primary cell/dura mater/skull/osteogenesis/Twist1/osteogenic differentiation分类
临床医学引用本文复制引用
安冉,邵国,张春阳..大鼠硬脑膜对颅骨成骨增强的影响[J].中国组织工程研究,2024,28(22):3478-3483,6.基金项目
国家自然科学基金(82160250,81960238),项目负责人:张春阳 （82160250,81960238）
国家临床重点专科建设项目经费资助,项目负责人:张春阳 （）
包头医学院研究生科研创新项目(bycx2023013),项目负责人:安冉 National Natural Science Foundation of China,No.82160250,81960238(to ZCY) （bycx2023013）
Funding for the National Clinical Key Specialty Construction Project(to ZCY) （to ZCY）
Graduate Student Research and Innovation Program of Baotou Medical College,No.bycx2023013(to AR) （to AR）
