Characterization of established cementoblast-like cell lines from human cementum-lining cells in vitro and in vivo

Kitagawa, Masae; Tahara, Hidetoshi; Kitagawa, Shoji; Oka, Hiroko; Kudo, Yasusei; Sato, Sunao; Ogawa, Ikuko; Miyaichi, Mutsumi; Takata, Takashi

doi:10.1016/j.bone.2006.05.022

Cited by 87 publications

(76 citation statements)

References 41 publications

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“…F-spondin (also known as spondin-1 encoded by SPON1) is an extracellular matrix protein that is required for the pathfinding of commissural axons during floor plate development (12,22). Higher expression of F-spondin enhanced that of osteocalcin and bone sialoprotein mRNA, as well as ALP in human periodontal ligament (HPL) cells, suggesting that F-spondin influences the differentiation of HPL cells (19,21). In the present study, the expression of SPON1 was weaker in compressed than in non-compressed HCEM.…”

Section: Discussionmentioning

confidence: 99%

“…An immortalized cell line of human cementoblasts (HCEM) was used in this study. The cells were provided by Professor Takashi Takata (Hiroshima University Graduate School), and their characteristics were previously reported (19,21). Cells were routinely maintained in alpha MEM (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich, St. Louis, MO, USA) and penicillin-streptomycin (100 IU/mL; Thermo Fisher Scientific) at 37°C in a humidified atmosphere of 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

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Response to light compressive force in human cementoblasts in vitro

et al. 2016

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The objective of this study is to investigate the responses of human cementoblasts to light compressive force in vitro. A human cementoblast cell line (HCEM) was loaded for 12 h by mounting coverslips (0.25 gf/cm 2 ). The coverslips were removed and the cells were cultured for up to 21 days. Cells without glass loading were used as controls. Cell growth, morphological changes, and the mRNA expression of RUNX2, ALP, WNT5A and SPON1 were investigated. No significant differences were observed in cell numbers between the compressed group and control group. Morphology of the compressed cells was slightly flattened on day 0; however, no indications of cell death were detected. Expression of differentiation markers including RUNX2, ALP and WNT5A was significantly lower in the compressed group (0.7, 0.75 and 0.75-fold respectively, P < 0.05) than in the control group on day 7. The expression levels of SPON1, a differentiation marker of cementoblasts, were higher on days 7 and 14 than on day 0, but were lower in the compressed group than in the control group (P < 0.01). These results suggest that light compressive force does not affect cell growth and morphology, but restrains higher expression of cementogenic differentiation markers in human cementoblasts in vitro.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Response to light compressive force in human cementoblasts in vitro

et al. 2016

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“…Human cementoblast-like (HCEM) cells have been established by transfection of hTERT (17). The cells express bone/ cementum-related proteins, such as type I collagen, runt-related transcription factor 2, osteocalcin, bone sialoprotein, and CP-23, show strong ALP activity and form calcified nodules.…”

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confidence: 99%

Brain-derived Neurotrophic Factor Stimulates Bone/Cementum-related Protein Gene Expression in Cementoblasts

Kajiya¹,

Shiba²,

Fujita³

et al. 2008

Journal of Biological Chemistry

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Brain-derived neurotrophic factor (BDNF), recognized as essential in the developing nervous system, is involved in differentiation and proliferation in non-neuronal cells, such as endothelial cells, osteoblasts, and periodontal ligament cells. We have focused on the application of BDNF to the regeneration of periodontal tissue and indicated that BDNF promotes the regeneration of experimentally created periodontal defects. Cementoblasts form cementum, mineralized tissue, which is key to establishing a functional periodontium. The application of BDNF to the regeneration of periodontal tissue requires elucidation of the mechanism by which BDNF regulates the functions of cementoblasts. In this study, we examined how BDNF regulates the mRNA expression of bone/cementum-related proteins (alkaline phosphatase (ALP), osteopontin (OPN), and bone morphogenetic protein-2 (BMP-2)) in cultures of immortalized human cementoblast-like (HCEM) cells. BDNF elevated the mRNA levels of ALP, OPN, and BMP-2 in HCEM cells. Small interfering RNA (siRNA) for TRKB, a high affinity receptor of BDNF, siRNA for ELK-1, which is a downstream target of ERK1/2, and PD98059, an ERK inhibitor, obviated the increase in the mRNA levels. BDNF increased the levels of phosphorylated ERK1/2 and Elk-1, and the blocking of BDNF signaling by treatment with siRNA for TRKB and PD98059 suppressed the phosphorylation of ERK1/2 and Elk-1. Furthermore, BDNF increased the levels of phosphorylated c-Raf, which activates the ERK signaling pathway. These findings provide the first evidence that the TrkB-c-Raf-ERK1/2-Elk-1 signaling pathway is required for the BDNF-induced mRNA expression of ALP, OPN, and BMP-2 in HCEM cells.

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“…Cell culture HCEM cells, immortalized by transfection of the hTERT gene, were established by Kitagawa et al (2006). The cells were cultured in a-minimum essential medium (a-MEM, Sigma, Steinheim, Germany) supplemented with 10% fetal bovine serum (FBS, GIBCO; Invitrogen, Buffalo, NY), penicillin G solution (100 U/ml), and streptomycin (100 mg/ml) (medium A).…”

Section: Methodsmentioning

confidence: 99%

Brain‐derived neurotrophic factor protects cementoblasts from serum starvation‐induced cell death

Kajiya

Shiba

Fujita

et al. 2009

Journal Cellular Physiology

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Our previous studies have shown that brain-derived neurotrophic factor (BDNF) enhances bone/cementum-related protein gene expression through the TrkB-c-Raf-ERK1/2-Elk-1 signaling pathway in cementoblasts, which play a critical role in the establishment of a functional periodontal ligament. To clarify how BDNF regulates survival in cementoblasts, we examined its effects on cell death induced by serum starvation in immortalized human cementoblast-like (HCEM) cells. BDNF inhibited the death of HCEM cells. Small-interfering RNA (siRNA) for TRKB, a high affinity receptor for BDNF, and for Bcl-2, countered the BDNF-induced decrease in dead cell number. In addition, LY294002, a PI3-kinase inhibitor; SH-6, an Akt inhibitor; and PDTC, a nuclear factor kappa B (NF-kappaB) inhibitor, but not PD98059, an ERK1/2 inhibitor, abolished the protective effect of BDNF against cell death. BDNF enhanced phosphorylated Akt levels, NF-kappaB activity in the nucleus, Bcl-2 mRNA levels, and mitochondrial membrane potential. The blocking of BDNF's actions by treatment with siRNA in all cases for TRKB and Bcl-2, LY294002, SH-6, and PDTC suppressed the enhancement. These findings provide the first evidence that a TrkB-PI3-kinase-Akt-NF-kappaB-Bcl-2 signaling pathway triggered by BDNF and the subsequent protective effect of BDNF on mitochondrial membrane potential are required to rescue HCEM cells from serum starvation-induced cell death. Furthermore, the survival and increased expression of bone/cementum-related proteins induced by BDNF in HCEM cells occur through different signaling pathways.

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Characterization of established cementoblast-like cell lines from human cementum-lining cells in vitro and in vivo

Cited by 87 publications

References 41 publications

<b>Response to light compressive force in human cementoblasts </b><i><b>in </b></i><i><b>vitro </b></i>

<b>Response to light compressive force in human cementoblasts </b><i><b>in </b></i><i><b>vitro </b></i>

Brain-derived Neurotrophic Factor Stimulates Bone/Cementum-related Protein Gene Expression in Cementoblasts

Brain‐derived neurotrophic factor protects cementoblasts from serum starvation‐induced cell death

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