Low-intensity Pulsed Ultrasound Stimulation Facilitates Osteogenic Differentiation of Human Periodontal Ligament Cells

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2014 Apr 19

PMID 24743551

Citations 33

Authors

Bo Hu

Yuanyuan Zhang

Jie Zhou

Jing Li

Feng Deng

Zhibiao Wang

Jinlin Song

Affiliations

Soon will be listed here.

Abstract

Human periodontal ligament cells (hPDLCs) possess stem cell properties, which play a key role in periodontal regeneration. Physical stimulation at appropriate intensities such as low-intensity pulsed ultrasound (LIPUS) enhances cell proliferation and osteogenic differentiation of mesechymal stem cells. However, the impacts of LIPUS on osteogenic differentiation of hPDLCs in vitro and its molecular mechanism are unknown. This study was undertaken to investigate the effects of LIPUS on osteogenic differentiation of hPDLCs. HPDLCs were isolated from premolars of adolescents for orthodontic reasons, and exposed to LIPUS at different intensities to determine an optimal LIPUS treatment dosage. Dynamic changes of alkaline phosphatase (ALP) activities in the cultured cells and supernatants, and osteocalcin production in the supernatants after treatment were analyzed. Runx2 and integrin β1 mRNA levels were assessed by reverse transcription polymerase chain reaction analysis after LIPUS stimulation. Blocking antibody against integrinβ1 was used to assess the effects of integrinβ1 inhibitor on LIPUS-induced ALP activity, osteocalcin production as well as calcium deposition. Our data showed that LIPUS at the intensity of 90 mW/cm2 with 20 min/day was more effective. The ALP activities in lysates and supernatants of LIPUS-treated cells started to increase at days 3 and 7, respectively, and peaked at day 11. LIPUS treatment significantly augmented the production of osteocalcin after day 5. LIPUS caused a significant increase in the mRNA expression of Runx2 and integrin β1, while a significant decline when the integrinβ1 inhibitor was used. Moreover, ALP activity, osteocalcin production as well as calcium nodules of cells treated with both daily LIPUS stimulation and integrinβ1 antibody were less than those in the LIPUS-treated group. In conclusion, LIPUS promotes osteogenic differentiation of hPDLCs, which is associated with upregulation of Runx2 and integrin β1, which may thus provide therapeutic benefits in periodontal tissue regeneration.

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References

Keselowsky B, Collard D, Garcia A . Integrin binding specificity regulates biomaterial surface chemistry effects on cell differentiation. Proc Natl Acad Sci U S A. 2005; 102(17):5953-7. PMC: 1087905. DOI: 10.1073/pnas.0407356102. View

Washio K, Iwata T, Mizutani M, Ando T, Yamato M, Okano T . Assessment of cell sheets derived from human periodontal ligament cells: a pre-clinical study. Cell Tissue Res. 2010; 341(3):397-404. DOI: 10.1007/s00441-010-1009-1. View

Salcedo L, Lian L, Jiang H, Sopko N, Penn M, Damaser M . Low current electrical stimulation upregulates cytokine expression in the anal sphincter. Int J Colorectal Dis. 2011; 27(2):221-5. DOI: 10.1007/s00384-011-1324-3. View

Lee N, Sowa H, Hinoi E, Ferron M, Ahn J, Confavreux C . Endocrine regulation of energy metabolism by the skeleton. Cell. 2007; 130(3):456-69. PMC: 2013746. DOI: 10.1016/j.cell.2007.05.047. View

Pitaru S, McCulloch C, Narayanan S . Cellular origins and differentiation control mechanisms during periodontal development and wound healing. J Periodontal Res. 1994; 29(2):81-94. DOI: 10.1111/j.1600-0765.1994.tb01095.x. View

Ciavarella S, Dammacco F, De Matteo M, Loverro G, Silvestris F . Umbilical cord mesenchymal stem cells: role of regulatory genes in their differentiation to osteoblasts. Stem Cells Dev. 2009; 18(8):1211-20. DOI: 10.1089/scd.2008.0340. View

Inubushi T, Tanaka E, Rego E, Kitagawa M, Kawazoe A, Ohta A . Effects of ultrasound on the proliferation and differentiation of cementoblast lineage cells. J Periodontol. 2008; 79(10):1984-90. DOI: 10.1902/jop.2008.080081. View

Wu P, Song J, Feng G, Dong N, Zhao C, Wang Z . [Repairing effects of low-intensity pulsed ultrasound on alveolar bone defects in Beagle dogs]. Hua Xi Kou Qiang Yi Xue Za Zhi. 2010; 28(5):522-5. View

Juliano R . Signal transduction by cell adhesion receptors and the cytoskeleton: functions of integrins, cadherins, selectins, and immunoglobulin-superfamily members. Annu Rev Pharmacol Toxicol. 2002; 42:283-323. DOI: 10.1146/annurev.pharmtox.42.090401.151133. View

10.

Fluck M . Functional, structural and molecular plasticity of mammalian skeletal muscle in response to exercise stimuli. J Exp Biol. 2006; 209(Pt 12):2239-48. DOI: 10.1242/jeb.02149. View

11.

Talic N, Evans C, Daniel J, George A, Zaki A . Immunohistochemical localization of alphavbeta3 integrin receptor during experimental tooth movement. Am J Orthod Dentofacial Orthop. 2004; 125(2):178-84. DOI: 10.1016/j.ajodo.2003.03.005. View

12.

Man J, Shelton R, Cooper P, Scheven B . Low-intensity low-frequency ultrasound promotes proliferation and differentiation of odontoblast-like cells. J Endod. 2012; 38(5):608-13. DOI: 10.1016/j.joen.2012.01.015. View

13.

Kanno T, Takahashi T, Tsujisawa T, Ariyoshi W, Nishihara T . Mechanical stress-mediated Runx2 activation is dependent on Ras/ERK1/2 MAPK signaling in osteoblasts. J Cell Biochem. 2007; 101(5):1266-77. DOI: 10.1002/jcb.21249. View

14.

Needleman I, Worthington H, Giedrys-Leeper E, Tucker R . Guided tissue regeneration for periodontal infra-bony defects. Cochrane Database Syst Rev. 2006; (2):CD001724. DOI: 10.1002/14651858.CD001724.pub2. View

15.

Yang Y, Rossi F, Putnins E . Periodontal regeneration using engineered bone marrow mesenchymal stromal cells. Biomaterials. 2010; 31(33):8574-82. DOI: 10.1016/j.biomaterials.2010.06.026. View

16.

Mostafa N, Uludag H, Dederich D, Doschak M, El-Bialy T . Anabolic effects of low-intensity pulsed ultrasound on human gingival fibroblasts. Arch Oral Biol. 2009; 54(8):743-8. DOI: 10.1016/j.archoralbio.2009.04.012. View

17.

Li L, Han M, Li S, Wang L, Xu Y . Cyclic tensile stress during physiological occlusal force enhances osteogenic differentiation of human periodontal ligament cells via ERK1/2-Elk1 MAPK pathway. DNA Cell Biol. 2013; 32(9):488-97. PMC: 3752521. DOI: 10.1089/dna.2013.2070. View

18.

Xue H, Zheng J, Cui Z, Bai X, Li G, Zhang C . Low-intensity pulsed ultrasound accelerates tooth movement via activation of the BMP-2 signaling pathway. PLoS One. 2013; 8(7):e68926. PMC: 3720872. DOI: 10.1371/journal.pone.0068926. View

19.

El-Bialy T, El-Shamy I, Graber T . Repair of orthodontically induced root resorption by ultrasound in humans. Am J Orthod Dentofacial Orthop. 2004; 126(2):186-93. DOI: 10.1016/j.ajodo.2004.02.010. View

20.

Yang Y, Yang Y, Li X, Cui L, Fu M, Rabie A . Functional analysis of core binding factor a1 and its relationship with related genes expressed by human periodontal ligament cells exposed to mechanical stress. Eur J Orthod. 2010; 32(6):698-705. DOI: 10.1093/ejo/cjq010. View