Quantitative comparison of parathyroid hormone 1 receptor between human periodontal ligament stem cells and human periodontal ligament cells

doi:10.3969/j.issn.2095-4344.2015.10.016

Abstract

Abstract:

BACKGROUND: Human periodontal ligament stem cells are a kind of mesenchymal stem cells that have self-renewal and multidifferentiation potential. Previous studies have showed that human periodontal ligament stem cells can differentiate into osteoblast-like cells or adipocyte-like cells under appropriate induction. Yet few studies have focused on the expression level of parathyroid hormone 1 receptor which will affect the osteogenic potential of Human periodontal ligament stem cells.

OBJECTIVE: To examine the expression level of parathyroid hormone 1 receptor between human periodontal ligament stem cells and human periodontal ligament cells and to discuss the role of parathyroid hormone 1 receptor in osteogenic differentiation.

METHODS: By using magnetic-bead cell sorting, we separated and identified the human periodontal ligament stem cells and human periodontal ligament cells. We examined and compared the mRNA expression level of parathyroid hormone 1 receptor in human periodontal ligament stem cells and human periodontal ligament cells by Real-Time PCR. Osteoblastic differentiation was examined through in vitro matrix mineralization by alizarin red staining and alkaline phosphatase assay.

RESULTS AND CONCLUSION: Positive immunomagetic sorted cells were positive for STRO-1, CD146, Vimentin, indicating that they were periodontal ligament stem cells. Parathyroid hormone 1 receptor was expressed in human periodontal ligament stem cells and mainly located in cell membrane and cytoplasm which were similar to human periodontal ligament cells and MG63 cells. The expression of parathyroid hormone 1 receptor in human periodontal ligament stem cells was 3.7 times higher than that in human periodontal ligament cells, which was similar to that in MG63 cells. After osteogenic induction, human periodontal ligament stem cells showed a higher expression of parathyroid hormone 1 receptor and osteoblast-related genes as well as the activity of osteoblast alkaline phosphatase and mineralization compared to human periodontal ligament cells. Our data showed that parathyroid hormone 1 receptor was higher in human periodontal ligament stem cells than human periodontal ligament cells and the expression was related with osteogenic differentiation, suggesting that human periodontal ligament stem cells display a higher potency of osteogenic differentiation and act as seed cells with a vast application prospect in oral tissue engineering.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: Stem Cells, Periodontal Ligament, Receptors, Parathyroid Hormone

CLC Number:

R394.2

Wang Yan-lan, Wang Xiao-xiao, Chen Tian-yu, Yang Fan-qiao, Dai Xu-bin, Ou Qian-min, Dai Shuang-ye, Lin Xue-feng . Quantitative comparison of parathyroid hormone 1 receptor between human periodontal ligament stem cells and human periodontal ligament cells [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(10): 1562-1569.

Figures/Tables 6

References

[1]Park JY, Jeon SH, Choung PH. Efficacy of periodontal stem cell transplantation in the treatment of advanced periodontitis. Cell Transplant. 2011;20(2):271-285.
[2]Yip I, Ma L, Mattheos N, et al. Defect healing with various bone substitutes. Clin Oral Implants Res. 2014. [Epub ahead of print]
[3]Goyal L. Clinical effectiveness of combining platelet rich fibrin with alloplastic bone substitute for the management of combined endodontic periodontal lesion. Restor Dent Endod. 2014;39(1):51-55.
[4]Bartold PM, McCulloch CA, Narayanan AS, et al. Tissue engineering: a new paradigm for periodontal regeneration based on molecular and cell biology. Periodontol 2000. 2000; 24:253-269.
[5]Maeda H, Tomokiyo A, Fujii S, et al. Promise of periodontal ligament stem cells in regeneration of periodontium. Stem Cell Res Ther. 2011;2(4):33.
[6]Ge S, Zhao N, Wang L, et al. Bone repair by periodontal ligament stem cellseeded nanohydroxyapatite-chitosan scaffold. Int J Nanomedicine. 2012;7:5405-5414.
[7]Seo BM, Miura M, Gronthos S, et al. Investigation of multipotent postnatal stem cells from human periodontal ligament. Lancet. 2004;364(9429):149-155.
[8]Jin SH, Lee JE, Yun JH, et al. Isolation and characterization of human mesenchymal stem cells from gingival connective tissue. J Periodontal Res. 2014. [Epub ahead of print]
[9]Ding G, Liu Y, Wang W, et al. Allogeneic periodontal ligament stem cell therapy for periodontitis in swine. Stem Cells. 2010; 28(10):1829-1838.
[10]Fawzy El-Sayed KM, Dörfer C, Fändrich F, et al. Adult mesenchymal stem cells explored in the dental field. Adv Biochem Eng Biotechnol. 2013;130:89-103.
[11]Cupp ME, Nayak SK, Adem AS, et al. Parathyroid hormone (PTH) and PTH-related peptide domains contributing to activation of different PTH receptor-mediated signaling pathways. J Pharmacol Exp Ther. 2013;345(3):404-418.
[12]Neer RM, Arnaud CD, Zanchetta JR, et al. Effect of parathyroid hormone (1-34) on fractures and bone mineral density in postmenopausal women with osteoporosis. N Engl J Med. 2001;344(19):1434-1441.
[13]Abuduwali N, Lossdörfer S, Winter J, et al. Functional characterization of the parathyroid hormone 1 receptor in human periodontal ligament cells. Clin Oral Investig. 2014; 18(2):461-470.
[14]Ahlström M, Pekkinen M, Lamberg-Allardt C. Dexamethasone downregulates the expression of parathyroid hormone-related protein (PTHrP) in mesenchymal stem cells. Steroids. 2009; 74(2):277-282.
[15]Wada N, Menicanin D, Shi S, et al. Immunomodulatory properties of human periodontal ligament stem cells. J Cell Physiol. 2009;219(3):667-676.
[16]Vasconcelos DF, Marques MR, Benatti BB, et al. Intermittent parathyroid hormone administration improves periodontal healing in rats. J Periodontol. 2014;85(5):721-728.
[17]Watson PH, Fraher LJ, Hendy GN, et al. Nuclear localization of the type 1 PTH/PTHrP receptor in rat tissues. J Bone Miner Res. 2000;15(6):1033-1044.
[18]Lupp A, Klenk C, Röcken C, et al. Immunohistochemical identification of the PTHR1 parathyroid hormone receptor in normal and neoplastic human tissues. Eur J Endocrinol. 2010; 162(5):979-986.
[19]Jong YJ, Sergin I, Purgert CA, et al. Location-dependent signaling of the group 1 metabotropic glutamate receptor mGlu5. Mol Pharmacol. 2014;86(6):774-785.
[20]O'Malley KL, Jong YJ, Gonchar Y, et al. Activation of metabotropic glutamate receptor mGlu5 on nuclear membranes mediates intranuclear Ca2+ changes in heterologous cell types and neurons. J Biol Chem. 2003; 278(30):28210-28219.
[21]Pickard BW, Hodsman AB, Fraher LJ, et al. Type 1 parathyroid hormone receptor (PTH1R) nuclear trafficking: association of PTH1R with importin alpha1 and beta. Endocrinology. 2006;147(7):3326-3332.
[22]Guo Y, Li Z, Ding R, et al. Parathyroid hormone induces epithelial-to-mesenchymal transition via the Wnt/β-catenin signaling pathway in human renal proximal tubular cells. Int J Clin Exp Pathol. 2014;7(9):5978-5987.
[23]Kraus D, Jäger A, Abuduwali N, et al. Intermittent PTH(1-34) signals through protein kinase A to regulate osteoprotegerin production in human periodontal ligament cells in vitro. Clin Oral Investig. 2012;16(2):611-618.
[24]Wolf M, Jäger A, Abuduwali N, et al. Continuous PTH modulates alkaline phosphatase activity in human PDL cells via protein kinase C dependent pathways in vitro. Ann Anat. 2013;195(5):455-460.
[25]Alokail MS, Peddie MJ. Quantitative comparison of PTH1R in breast cancer MCF7 and osteosarcoma SaOS-2 cell lines. Cell Biochem Funct. 2008;26(4):522-533.
[26]Gentili C, Morelli S, de Boland AR. Characterization of PTH/PTHrP receptor in rat duodenum: effects of ageing. J Cell Biochem. 2003;88(6):1157-1167.
[27]Zhou S, Bueno EM, Kim SW, et al. Effects of age on parathyroid hormone signaling in human marrow stromal cells. Aging Cell. 2011;10(5):780-788.
[28]Swarthout JT, D'Alonzo RC, Selvamurugan N, et al. Parathyroid hormone-dependent signaling pathways regulating genes in bone cells. Gene. 2002;282(1-2):1-17.
[29]Luck MD, Carter PH, Gardella TJ. The (1-14) fragment of parathyroid hormone (PTH) activates intact and amino-terminally truncated PTH-1 receptors. Mol Endocrinol. 1999;13(5):670-680.
[30]Blind E, Raue F, Knappe V, et al. Cyclic AMP formation in rat bone and kidney cells is stimulated equally by parathyroid hormone-related protein (PTHrP) 1-34 and PTH 1-34. Exp Clin Endocrinol. 1993;101(3):150-155.
[31]Potetinova Z, Barbier JR, Suen T, et al. C-terminal analogues of parathyroid hormone: effect of C-terminus function on helical structure, stability, and bioactivity. Biochemistry. 2006;45(37):11113-11121.
[32]Pioszak AA, Xu HE. Molecular recognition of parathyroid hormone by its G protein-coupled receptor. Proc Natl Acad Sci U S A. 2008;105(13):5034-5039.
[33]Datta NS, Abou-Samra AB. PTH and PTHrP signaling in osteoblasts. Cell Signal. 2009;21(8):1245-1254.
[34]Esbrit P, Alcaraz MJ. Current perspectives on parathyroid hormone (PTH) and PTH-related protein (PTHrP) as bone anabolic therapies. Biochem Pharmacol. 2013;85(10): 1417-1423.
[35]Sidiropoulou-Chatzigiannis S, Kourtidou M, Tsalikis L.The effect of osteoporosis on periodontal status, alveolar bone and orthodontic tooth movement. A literature review. J Int Acad Periodontol. 2007;9(3):77-84.
[36]Qiu T, Xian L, Crane J, et al. PTH receptor signaling in osteoblasts regulates endochondral vascularization in maintenance of postnatal growth plate. J Bone Miner Res. 2015;30(2):309-317.
[37]Haramoto N, Kawane T, Horiuchi N. Upregulation of PTH receptor mRNA expression by dexamethasone in UMR-106 osteoblast-like cells. Oral Dis. 2007;13(1):23-31.
[38]Cochran DL, Jones A, Heijl L, et al. Periodontal regeneration with a combination of enamel matrix proteins and autogenous bone grafting.J Periodontol. 2003;74(9):1269-1281.
[39]Gao Y, Zhao G, Li D, et al. Isolation and multiple differentiation potential assessment of human gingival mesenchymal stem cells. Int J Mol Sci. 2014;15(11):20982-20996.
[40]Han J, Menicanin D, Marino V, et al. Assessment of the regenerative potential of allogeneic periodontal ligament stem cells in a rodent periodontal defect model. J Periodontal Res. 2014;49(3):333-345.
[41]Trubiani O, Orsini G, Zini N, et al. Regenerative potential of human periodontal ligament derived stem cells on three-dimensional biomaterials: a morphological report. J Biomed Mater Res A. 2008;87(4):986-993.