Journal of Shanghai Jiao Tong University (Medical Science) >

2022 , Vol. 42 >Issue 8: 1008 - 1015

DOI: https://doi.org/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

  • Xijun WANG ,
  • Anting JIN ,
  • Xiangru HUANG ,
  • Hongyuan XU ,
  • Xin GAO ,
  • Yiling YANG ,
  • Qinggang DAI ,
  • Lingyong JIANG
Expand
  • 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
JIANG Lingyong, E-mail: jianglingyong@sjtu.edu.cn.

Received date: 2022-04-14

  Accepted date: 2022-08-13

  Online published: 2022-10-08

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)

Fold

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-CreERT2mice were mated with R26tdTomato 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-CreERT2;R26tdTomato . 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-CreERT2;R26tdTomato 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

Cite this article

Xijun WANG , Anting JIN , Xiangru HUANG , Hongyuan XU , Xin GAO , Yiling YANG , Qinggang DAI , Lingyong JIANG . 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 . DOI: 10.3969/j.issn.1674-8115.2022.08.005

References

1 LI Y N, JACOX L A, LITTLE S H, et al. Orthodontic tooth movement: the biology and clinical implications[J]. Kaohsiung J Med Sci, 2018, 34(4): 207-214.
2 SEO B M, MIURA M, GRONTHOS S, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament[J]. Lancet, 2004, 364(9429): 149-155.
3 WISE G E, KING G J. Mechanisms of tooth eruption and orthodontic tooth movement[J]. J Dent Res, 2008, 87(5): 414-434.
4 赵闫, 欧阳宁鹃, 代杰文, 等. 细胞谱系示踪技术及其在口腔医学研究中的应用进展[J].口腔生物医学, 2017, 8(4): 205-210.
4 ZHAO Y, OUYANG N J, DAI J W, et al. Development of cell lineage tracing technology and its application in stomatology research[J]. Oral Biomed, 2017, 8(4): 205-210.
5 BULTE J W M, DOUGLAS T, WITWER B, et al. Monitoring stem cell therapy in vivo using magnetodendrimers as a new class of cellular MR contrast agents[J]. Acad Radiol, 2002, 9(Suppl 2): S332-S335.
6 NAGY A. Cre recombinase: the universal reagent for genome tailoring[J]. Genesis, 2000, 26(2): 99-109.
7 SHI Y, HE G X, LEE W C, et al. Gli1 identifies osteogenic progenitors for bone formation and fracture repair[J]. Nat Commun, 2017, 8(1): 2043.
8 YI Y, STENBERG W, LUO W, et al. Alveolar bone marrow Gli1+ stem cells support implant osseointegration[J]. J Dent Res, 2022, 101(1): 73-82.
9 MIZOGUCHI T, PINHO S, AHMED J, et al. Osterix marks distinct waves of primitive and definitive stromal progenitors during bone marrow development[J]. Dev Cell, 2014, 29(3): 340-349.
10 KAWANAMI A, MATSUSHITA T, CHAN Y Y, et al. Mice expressing GFP and CreER in osteochondro progenitor cells in the periosteum[J]. Biochem Biophys Res Commun, 2009, 386(3): 477-482.
11 BASSIR S H, GARAKANI S, WILK K, et al. Prx1 expressing cells are required for periodontal regeneration of the mouse incisor[J]. Front Physiol, 2019, 10: 591.
12 GONG X Y, ZHANG H, XU X Q, et al. Tracing PRX1+ cells during molar formation and periodontal ligament reconstruction[J]. Int J Oral Sci, 2022, 14(1): 5.
13 ZHONG Z A, SUN W H, CHEN H Y, et al. Optimizing tamoxifen-inducible Cre/Loxp system to reduce tamoxifen effect on bone turnover in long bones of young mice[J]. Bone, 2015, 81: 614-619.
14 DAI Q G, ZHOU S R, ZHANG P, et al. Force-induced increased osteogenesis enables accelerated orthodontic tooth movement in ovariectomized rats[J]. Sci Rep, 2017, 7(1): 3906.
15 KUSUMBE A P, RAMASAMY S K, STARSICHOVA A, et al. Sample preparation for high-resolution 3D confocal imaging of mouse skeletal tissue[J]. Nat Protoc, 2015, 10(12): 1904-1914.
16 孙聪, 刘文佳, 金岩, 等. 小鼠正畸牙齿移动模型的制作[J]. 牙体牙髓牙周病学杂志, 2017, 27(5): 280-283.
16 SUN C, LIU W J, JIN Y, et al. Establishment of mouse model of orthodontic tooth movement[J]. Chin J Conserv Dent, 2017, 27(5): 280-283.
17 ABDALLAH B M, KASSEM M. Human mesenchymal stem cells: from basic biology to clinical applications[J]. Gene Ther, 2008, 15(2): 109-116.
18 WANG K, XU C, XIE X, et al. Axin2+ PDL cells directly contribute to new alveolar bone formation in response to orthodontic tension force[J]. J Dent Res, 2022, 101(6): 695-703.
19 KIM H, KIM M, IM S K, et al. Mouse Cre-LoxP system: general principles to determine tissue-specific roles of target genes[J]. Lab Anim Res, 2018, 34(4): 147-159.
20 FEIL R, BROCARD J, MASCREZ B, et al. Ligand-activated site-specific recombination in mice[J]. Proc Natl Acad Sci U S A, 1996, 93(20): 10887-10890.
21 XIE X, WANG J, WANG K, et al. Axin2+-mesenchymal PDL cells, instead of K14+ epithelial cells, play a key role in rapid cementum growth[J]. J Dent Res, 2019, 98(11): 1262-1270.
22 LI B, WANG Y G, FAN Y, et al. Cranial suture mesenchymal stem cells: insights and advances[J]. Biomolecules, 2021, 11(8): 1129.
23 MARTIN J F, BRADLEY A, OLSON E N. The paired-like homeo box gene MHox is required for early events of skeletogenesis in multiple lineages[J]. Genes Dev, 1995, 9(10): 1237-1249.
24 LU M F, CHENG H T, KERN M J, et al. Prx-1 functions cooperatively with another paired-related homeobox gene, prx-2, to maintain cell fates within the craniofacial mesenchyme[J]. Development, 1999, 126(3): 495-504.
25 MITCHELL J M, HICKLIN D M, DOUGHTY P M, et al. The Prx1 homeobox gene is critical for molar tooth morphogenesis[J]. J Dent Res, 2006, 85(10): 888-893.
26 BALIC A, ADAMS D, MINA M N. Prx1 and Prx2 cooperatively regulate the morphogenesis of the medial region of the mandibular process[J]. Dev Dyn, 2009, 238(10): 2599-2613.
27 WANG J, SARASWAT D, SINHA A K, et al. Paired related homeobox protein 1 regulates quiescence in human oligodendrocyte progenitors[J]. Cell Rep, 2018, 25(12): 3435-3450.e6.
Options
Outlines

/

〈 〉