Col1+ 细胞在牙槽窝愈合中的时空分布及分化潜能

doi:10.3969/j.issn.1674-8115.2026.02.002

上海交通大学学报（医学版） ›› 2026, Vol. 46 ›› Issue (2): 143-150.doi: 10.3969/j.issn.1674-8115.2026.02.002

• 论著 · 基础研究 • 上一篇

Col1⁺ 细胞在牙槽窝愈合中的时空分布及分化潜能

赵昱¹, 昝冰欣¹, 孙思远², 黄湘如², 高洁³, 吴轶群³(), 江凌勇²(), 代庆刚³()

^1.滨州医学院口腔医学院，烟台 264003
^2.上海交通大学医学院附属第九人民医院口腔颅颌面科，上海交通大学口腔医学院，国家口腔医学中心，国家口腔疾病临床医学研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011
^3.上海交通大学医学院附属第九人民医院口腔第二门诊部，上海交通大学口腔医学院，国家口腔医学中心，国家口腔疾病临床医学研究中心，上海市口腔医学重点实验室，上海市口腔医学研究所，上海 200011

收稿日期:2025-05-05 接受日期:2025-12-19 出版日期:2026-02-28 发布日期:2026-02-28
通讯作者: 吴轶群，主任医师，博士；电子信箱：yiqunwu@hotmail.com。
江凌勇，主任医师，博士；电子信箱：jianglingyong@sjtu.edu.cn。
代庆刚，副主任医师，博士；电子信箱：daiqinggang@sjtu.edu.cn。
作者简介:第一联系人：为共同第一作者（*co-first authors）。
基金资助:
上海交通大学医学院“双百人”项目(20221809);国家自然科学基金(82430032,82071083,82271006,82271004,82471007);国家重点研发计划(2024YFC2510700);上海市自然科学基金(22ZR1436700,21ZR1436900,21ZR1437700,24ZR1491900,25ZR1401217);上海交通大学医学院附属第九人民医院交叉研究基金(JYJC202116,JYJC202411);上海市科技创新行动计划国际科技合作项目/政府间国际科技合作项目(23410713600);海南省自然科学基金(824MS152);海南健康科学与技术协同创新联合项目(WSJK2025MS196)

Spatiotemporal distribution and differentiation potential of Col1⁺ cells in alveolar socket healing

Zhao Yu¹, Zan Bingxin¹, Sun Siyuan², Huang Xiangru², Gao Jie³, Wu Yiqun³(), Jiang Lingyong²(), Dai Qinggang³()

^1.Binzhou Medical University School of Stomatology, Yantai 264003, China
^2.Department of Oral and Maxillofacial Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology, Shanghai 200011, China
^3.Department of Second Dental Centre, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; College of Stomatology, Shanghai Jiao Tong University; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology; Shanghai Research Institute of Stomatology, Shanghai 200011, China

Received:2025-05-05 Accepted:2025-12-19 Online:2026-02-28 Published:2026-02-28
Contact: Wu Yiqun, E-mail: yiqunwu@hotmail.com.
Jiang Lingyong, E-mail: jianglingyong@sjtu.edu.cn.
Dai Qinggang, E-mail: daiqinggang@sjtu.edu.cn.
Supported by:
"Two-hundred Talents" Program of Shanghai Jiao Tong University School of Medicine(20221809);National Natural Science Foundation of China(82430032,82071083,82271006,82271004,82471007);National Key Research and Development Program of China(2024YFC2510700);Natural Science Foundation of Shanghai(22ZR1436700,21ZR1436900,21ZR1437700,24ZR1491900,25ZR1401217);Cross-disciplinary Research Fund of Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine(JYJC202116,JYJC202411);International Science and Technology Cooperation Project of Shanghai Science and Technology Innovation Action Plan/Inter-Governmental International Science and Technology Cooperation Project(23410713600);Natural Science Foundation of Hainan Province(824MS152);Joint Program on Health Science & Technology Innovation of Hainan(WSJK2025MS196)

摘要/Abstract

摘要：

目的·将Cre/loxP重组酶系统与拔牙模型相结合，探究分泌Ⅰ型胶原蛋白（collagen type Ⅰ，COL1）的Col1⁺ 细胞在牙槽窝愈合过程中的时空分布规律及其向成骨细胞的分化潜能。方法·将Col1-CreERT2小鼠与Rosa26-LoxP-Stop-LoxP-tdTomato小鼠交配繁殖，以获得Col1-CreERT2；tdTomato双转基因子代小鼠。取15只子代小鼠，通过腹腔注射他莫昔芬（tamoxifen，TA）标记Col1⁺ 细胞谱系，1周后于全身麻醉下完整拔除子代小鼠右侧上颌第一磨牙，以构建双转基因小鼠拔牙模型。将子代小鼠分为5组（每组3只），分别在0、3、7、14、28 d取小鼠上颌骨样本，通过石蜡切片观察Col1⁺ 细胞谱系在牙槽窝愈合过程中的动态分布；利用免疫荧光技术标记成骨细胞特异性转录因子（osterix，OSX）、血管标志物内皮黏蛋白（endomucin，EMCN）、神经元标志物β3微管蛋白（β3-tubulin）探究Col1⁺ 细胞的成骨潜能及其与血管、神经的空间定位关系。结果·成功构建了Col1-CreERT2；tdTomato双转基因小鼠拔牙模型。石蜡切片观察的结果显示：建模后第3日，牙槽窝中出现Col1⁺细胞；第7日，Col1⁺ 细胞的数量增加，牙槽窝逐渐成骨；第28日，Col1⁺ 细胞的比例进一步上升，且分布于骨小梁周围。统计结果显示，Col1⁺ 细胞数量在牙槽窝愈合过程中随时间的推移而增加（与第0日相比，均P<0.001）。免疫荧光技术检测的结果显示，建模后第7~28日，牙槽窝内Col1⁺OSX⁺双阳性细胞数量逐渐增多，Col1⁺ 细胞在空间定位上分别与EMCN和β3-tubulin相邻。结论·在牙槽窝愈合过程中，Col1⁺ 细胞具有向成骨细胞分化的潜能，还可能参与血管、神经生成。

关键词: 谱系示踪技术, Ⅰ型胶原蛋白, Cre/loxP重组酶系统, 牙槽窝愈合

Abstract:

Objective ·To combine the Cre/loxP recombinase system with a tooth extraction model to explore the spatiotemporal distribution of Col1⁺ cells secreting type Ⅰ collagen (collagen type Ⅰ, COL1) during alveolar socket healing and their potential to differentiate into osteoblasts. Methods ·Col1-CreERT2 mice were mated with Rosa26-LoxP-Stop-LoxP-tdTomato mice to obtain Col1-CreERT2;tdTomato double transgenic offspring mice. Fifteen offspring mice were selected and tamoxifen (TA) was intraperitoneally injected to label the Col1⁺ cell lineage. One week later, the right maxillary first molars of the offspring mice were completely extracted under general anesthesia to establish a double transgenic mouse tooth extraction model. The offspring mice were divided into 5 groups, with 3 mice in each group. Maxillary bone samples were collected at 0, 3, 7, 14, and 28 days after tooth extraction, respectively. Paraffin sections were used to observe the dynamic distribution of the Col1⁺ cell lineage during alveolar socket healing. Immunofluorescence techniques were employed to label the osteoblast-specific transcription factor osterix (OSX), vascular marker endomucin (EMCN), and neuronal marker β3-tubulin to investigate the osteogenic potential of Col1⁺ cells and their spatial localization relationship with blood vessels and nerves. Results ·The tooth extraction model of Col1-CreERT2;tdTomato double transgenic mice was successfully established. Paraffin section observations revealed that Col1⁺ cells appeared in the alveolar socket on the third day after the model was established; on the seventh day, the number of Col1⁺ cells increased and alveolar socket osteogenesis gradually occurred; on the 28th day, the proportion of Col1⁺ cells further increased and they were distributed around the bone trabeculae. The results of statistical analysis showed that the number of Col1⁺ cells increased over time during alveolar socket healing (all P<0.001, compared with day 0). The results of Immunofluorescence assay indicated that the number of Col1⁺ OSX⁺ double-positive cells in the alveolar socket gradually increased from the 7th to the 28th day after the model was established, and Col1⁺ cells were spatially adjacent to EMCN and β3-tubulin. Conclusion ·During alveolar socket healing, Col1⁺ cells have the potential to differentiate into osteoblasts, and may also be involved in the formation of blood vessels and nerves.

Key words: lineage tracing technology, collagen type Ⅰ (COL1), Cre/loxP recombinase system, alveolar socket healing

中图分类号:

R782.11

赵昱, 昝冰欣, 孙思远, 黄湘如, 高洁, 吴轶群, 江凌勇, 代庆刚. Col1⁺ 细胞在牙槽窝愈合中的时空分布及分化潜能[J]. 上海交通大学学报（医学版）, 2026, 46(2): 143-150.

Zhao Yu, Zan Bingxin, Sun Siyuan, Huang Xiangru, Gao Jie, Wu Yiqun, Jiang Lingyong, Dai Qinggang. Spatiotemporal distribution and differentiation potential of Col1⁺ cells in alveolar socket healing[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2026, 46(2): 143-150.

图/表 6

表 1 PCR引物序列

Tab 1 Primer sequences for PCR

Gene	Forward sequence (5'→3')	Reverse sequence (5'→3')
Col1-CreERT2	CGATGCAACGAGTGATGAGG	CGCATAACCAGTGAAACAGC
tdTomato	TTCGGCTTCTGGCGTGTGAC	GAAGAGGAGTGCTCCGATCAGG

图1 双转基因子代小鼠拔牙模型构建及其样本获取流程图

Fig 1 Flowchart of the construction and sample collection of the tooth extraction model in double transgenic offspring mice

图2 双转基因子代小鼠的基因型鉴定与拔牙模型构建Note： A/B. Genotype identification of the offspring mice for the tdTomato gene (A) and Cre gene (B). tdTomato represents Rosa26-LoxP-Stop-LoxP-tdTomato, and Cre represents Col1-CreERT2. C. Schematic diagram of different states of Col1-CreERT2 cells under conditions with or without TA. ER—estrogen receptor. D. Construction of the tooth extraction model in the offspring mice. The arrows indicate the fresh alveolar socket.

Fig 2 Genotype identification of double transgenic offspring mice and establishment of the tooth extraction model

图3 牙槽窝愈合过程中 Col1+ 细胞的时空分布Note： A. Detection of the quantity and distribution of Col1+ cells during the healing process of the alveolar socket by immunofluorescence assay (×20). The tdTomato+ cells are stained red, and the blue signal represents the result after staining with DAPI. The dotted lines show the edge of the alveolar socket. B. Statistical analysis of the percentage of Col1+ cells in the process of alveolar socket healing. ①P<0.001, compared with the Day 0 group.

Fig 3 Spatiotemporal distribution of Col1+ cells during the healing process of the alveolar socket

图 4 牙槽窝愈合过程中 Col1+ 细胞与OSX+ 细胞的时空分布Note： A. Detection of the distribution of Col1+ and OSX+ cells in the alveolar socket at different time points by immunofluorescence assay (above ×20, below×40). The tdTomato+ cells are stained red, while the blue color represents the result after DAPI staining. The green color marks the OSX+ cells, and the OSX+ tdTomato+ double-positive cells appear yellow. The dotted lines show the edge of the alveolar socket. B. Statistical analysis of the proportion of double-positive cells (OSX+ and tdTomato+) in the total tdTomato+ cells and total OSX+ cells during the period of alveolar socket healing.

Fig 4 Spatiotemporal distribution of Col1+ cells and OSX+ cells during the healing process of the alveolar socket

图5 免疫荧光技术检测 Col1+ 细胞与EMCN+ 细胞（A）以及与β3-tubulin+ 细胞（B）在牙槽窝内的分布情况及空间关联性Note： The green areas respectively mark EMCN and β3-tubulin. The dotted lines represent the edge of the alveolar socket. In figures A and B, the upper panels ×20, and the lower panels ×40.

Fig 5 Detection of the distribution and spatial correlation of Col1+ /EMCN+ cells (A) and Col1+ /β3-tubulin+ cells (B) in alveolar sockets by immunofluorescence assay

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Col1⁺ 细胞在牙槽窝愈合中的时空分布及分化潜能

Spatiotemporal distribution and differentiation potential of Col1⁺ cells in alveolar socket healing

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Col1+ 细胞在牙槽窝愈合中的时空分布及分化潜能

Spatiotemporal distribution and differentiation potential of Col1+ cells in alveolar socket healing

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Col1⁺ 细胞在牙槽窝愈合中的时空分布及分化潜能

Spatiotemporal distribution and differentiation potential of Col1⁺ cells in alveolar socket healing