Study on Prrx1+ periodontal ligament stem cells during orthodontic tooth movement by lineage tracing

doi:10.3969/j.issn.1674-8115.2022.08.005

Journal of Shanghai Jiao Tong University (Medical Science) ›› 2022, Vol. 42 ›› Issue (8): 1008-1015.doi: 10.3969/j.issn.1674-8115.2022.08.005

• Basic research •

Study on Prrx1⁺ periodontal ligament stem cells during orthodontic tooth movement by lineage tracing

WANG Xijun¹(), JIN Anting¹(), HUANG Xiangru¹, XU Hongyuan¹, GAO Xin¹, YANG Yiling¹, DAI Qinggang², JIANG Lingyong¹()

^1.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 200011, China
^2.The 2nd Dental Centre, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine; National Center for Stomatology; National Clinical Research Center for Oral Diseases; Shanghai Key Laboratory of Stomatology, Shanghai 200011, China

Received:2022-04-14 Accepted:2022-08-13 Online:2022-08-28 Published:2022-10-08
Contact: JIANG Lingyong E-mail:838657340@qq.com;Katerinajin@foxmail.com;jianglingyong@sjtu.edu.cn
Supported by:
National Natural Science Foundation of China(82071083);Natural Science Foundation of Shanghai(22ZR1436700);Program of Shanghai Academic/Technology Research Leader(20XD1422300);Cross-disciplinary Research Fund of Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine(JYJC202116);Project of Biobank of Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine(YBKB201919);Clinical Research Plan of Shanghai Hospital Development Center(SHDC2020CR4084);Students Innovation Training Program of Shanghai Jiao Tong University School of Medicine(S202110248114)

Abstract

Abstract:

Objective ·To investigate the dynamic distribution of paired related homebox 1 (Prrx1) positive cell lineage (Prrx1⁺ cell lineage) in periodontal ligament stem cells (PDLSCs) during tooth movement in mice by Cre/loxP recombination system. Methods ·By using Cre/loxP recombination system, inducible Prrx1-Cre^ERT2^{mice were mated with R26^tdTomato fluorescently labeledmice, and their offspring mice were genotyped by PCR. Eight offspring mice were injected with tamoxifen (TA) intraperitoneally to mark the Prrx1⁺^{cell lineage (tdTomato⁺ cells) in PDLSCs. The orthodontics tooth movement (OTM) model was constructed by placing a force-loading device on the left side of maxilla (i.e., OTM side), and the right side without force was control side (i.e., Ctrl side). The mice were sacrificed on the third day (OTM 3 d) and the seventh day (OTM 7 d) of tooth movement, and their bilateral maxillary molars and surrounding periodontium were collected, decalcified, embedded and frozen sectioned. The changes of periodontal ligament in the tension area and compression area were observed by hematoxylin-eosin staining (H-E staining), and the dynamic distribution of Prrx1⁺ celllineage was observed by immunofluorescence staining. Results ·The genotype of the offspring mice was identified by PCR as Prrx1-Cre^ERT2;R26^tdTomato . With the increased action of the stressing device, the tooth movement distance of OTM 7 d mice [(87.44±4.02) μm] increased significantly compared with that of the OTM 3 d mice [(42.81±5.04) μm], suggesting OTM model was successfully constructed. H-E staining showed that the periodontal ligament and its gap in the compression area of the OTM side was compressed and narrowed on OTM 3 d, and its width was gradually restored on OTM 7 d; the periodontal ligament in the tension area of the OTM side was stretched on OTM 3 d, and the periodontal ligament was more regularly arranged on OTM 7 d than that on OTM 3 d. Immunofluorescence staining showed that the number of tdTomato⁺ cells in the periodontal ligament of the compression area on the OTM side was lower than that of the Ctrl side on OTM 3 d, and the number of tdTomato⁺ cells in compression area on the OTM side was higher than that of the Ctrl side on OTM 7 d (both P<0.05); on both OTM 3 d and OTM 7 d, the number of tdTomato⁺ cells in tension area on the OTM side increased compared with that on the Ctrl side (both P<0.05). Conclusion ·The OTM model of Prrx1-Cre^ERT2;R26^tdTomato mice is successfully constructed, and the involvement of Prrx1⁺ cell lineage in periodontal remodeling during OTM is tentatively confirmed.}}

Key words: cell lineage tracing, orthodontic tooth movement (OTM), paired related homeobox1 (Prrx1), Cre/loxP recombinase system

CLC Number:

R783.5

WANG Xijun, JIN Anting, HUANG Xiangru, XU Hongyuan, GAO Xin, YANG Yiling, DAI Qinggang, JIANG Lingyong. Study on Prrx1⁺ periodontal ligament stem cells during orthodontic tooth movement by lineage tracing[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022, 42(8): 1008-1015.

Figures/Tables 5

Tab 1 Primer sequences for PCR

Gene	Forward sequence （5'→3'）	Reverse sequence （5'→3'）
Prrx1-Cre^ERT2	CGATGCAACGAGTGATGAGG	CGCATA ACCAGTGAAACAGC
tdTomato	TTCGGCTTCTGGCGTGTGAC	GAAGAGGAGTGCTCCGATCAGG
tdTomato-WT	AGCACTTGCTCTCCCAAAGTCGCTC	AAACTTCCCGACAAAACCGAA

Fig 1 Detection of genotype of offspring mice by PCR

Fig 2 Construction and validation of OTM model in offspring mice

Fig 3 Changes of periodontal ligament and expression of Prrx1+ cell lineage in the compression area

Fig 4 Changes of periodontal ligament and expression of Prrx1+ cell lineage in the tension area

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References 27

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Study on Prrx1⁺ periodontal ligament stem cells during orthodontic tooth movement by lineage tracing

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