Role of chondroitin sulfate in the developmental and healing process of the dental pulp in mice
Role of chondroitin sulfate in the developmental and healing process of the dental pulp in mice
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硫酸软骨素在小鼠牙髓发育和愈合过程中的作用
DOI：
10.1007/s00441-022-03575-3
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Hayato Ohshima
中科院分区：
文献类型：
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作者：
Hiroko Ida-Yonemochi ;Kosei Takeuchi ;Hayato Ohshima
硫酸软骨素蛋白聚糖（CSPG）是一种重要的细胞外基质，在器官形成过程中起着重要作用。其核心蛋白和硫酸软骨素（CS）链具有特定的生物学功能。为了阐明CS在牙髓发育和愈合过程中的作用，我们在CS N-乙酰氨基半乳糖转移酶-1（T1）基因敲除（KO）小鼠中进行了实验性牙齿再植。我们还使用T1-siRNA技术和外部CS对WT和T1 KO原代牙髓细胞进行细胞增殖测定和qRT-PCR分析。在牙齿发育过程中，CS在牙乳头中呈弥漫性表达，随着牙髓的成熟，CS在分化区（包括成牙本质细胞）中消失。在完全发育的磨牙，CS被限制在根尖区域与Gli 1阳性细胞共定位。在再植牙愈合过程中，CD 31阳性细胞在WT磨牙的CS阳性基质中积累。在T1 KO磨牙中，在愈合早期，牙髓中Ki 67-和Gli 1-阳性细胞的出现明显少于WT磨牙，并且在T1 KO小鼠中I型胶原阳性修复性牙本质形成不明显。在原代培养实验中，T1基因的siRNA敲低显著抑制WT牙髓细胞的增殖，外源性CS显著上调T1 KO牙髓细胞中cyclin D1和CD 31的mRNA表达。这些结果表明，CS是参与细胞增殖和功能分化的牙髓组成细胞，包括血管细胞，在牙齿损伤后的牙髓组织愈合过程中。
Chondroitin sulfate proteoglycan (CSPG), one of the major extracellular matrices, plays an important part in organogenesis. Its core protein and chondroitin sulfate (CS) chain have a specific biological function. To elucidate the role of CS in the developmental and healing process of the dental pulp, we performed an experimental tooth replantation in CS N-acethylgalactosaminyltransferase-1 (T1) gene knockout (KO) mice. We also performed cell proliferation assay and qRT-PCR analysis for the WT and T1KO primary dental pulp cells using T1-siRNA technique and external CS. During tooth development, CS was diffusely expressed in the dental papilla, and with dental pulp maturation, CS disappeared from the differentiated areas, including the odontoblasts. In fully developed molars, CS was restricted to the root apex region colocalizing with Gli1-positive cells. In the healing process after tooth replantation, CD31-positive cells accumulated in the CS-positive stroma in WT molars. In T1KO molars, the appearance of Ki67- and Gli1-positive cells in the dental pulp was significantly fewer than in WT molars in the early healing stage, and collagen I-positive reparative dentin formation was not obvious in T1KO mice. In primary culture experiments, siRNA knockdown of T1 gene significantly suppressed cell proliferation in WT dental pulp cells, and the mRNA expression ofcyclin D1andCD31was significantly upregulated by external CS in T1KO dental pulp cells. These results suggest that CS is involved in the cell proliferation and functional differentiation of dental pulp constituent cells, including vascular cells, in the healing process of dental pulp tissue after tooth injury.