Oligomerization-induced Conformational Change in the C-terminal Region of Nel-like Molecule 1 (NELL1) Protein Is Necessary for the Efficient Mediation of Murine MC3T3-E1 Cell Adhesion and Spreading

Nakamura, Yoko; Hasebe, Ai; Takahashi, Kaneyoshi; Iijima, Masumi; Yoshimoto, Nobuo; Maturana, Andrés D.; Ting, Kang; Kuroda, Shinya; Niimi, Tomoaki

doi:10.1074/jbc.m113.507020

Cited by 15 publications

(18 citation statements)

References 45 publications

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“…The results showed that Nell-1 effectively activated the ERK and JNK pathways rather than the P38 pathway in pre-osteoblasts on Ti surfaces. Nakamura Y et al found that Nell-1 mediated cell adhesion by binding to integrin receptors to activate ERK in MC3T3-E1 cells [47], confirming that Nell-1 can activate the ERK pathway in pre-osteoblasts. Interestingly, the selective activation of different MAPK pathways by Nell-1 depends on cell type [26].…”

Section: Discussionmentioning

confidence: 86%

Nell-1 Enhances Osteogenic Differentiation of Pre-Osteoblasts on Titanium Surfaces via the MAPK-ERK Signaling Pathway

Shen

Wang

et al. 2018

Cell Physiol Biochem

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Background/Aims: This study aimed to investigate the effect of Nell-1 on the osteogenic behaviors of pre-osteoblasts on titanium (Ti) surfaces and to identify the underlying signaling pathway. Methods: Nell-1 at different concentrations was added to culture medium to stimulate MC3T3-E1 subclone 14 on Ti surfaces. A CCK-8 colorimetric assay was used to detect cell proliferation. Alkaline phosphatase activity (ALP) assay and enzyme-linked immunosorbent assay (ELISA) were used to evaluate ALP activity and the osteocalcin (OCN) secretion, respectively. Indicators of osteoblastic differentiation were assessed using real-time polymerase chain reaction analysis (RT-PCR). Western blot (WB) assay was used to analyze the expression changes of the osteogenic proteins and the mitogen-activated protein kinase (MAPK) pathway. Results: Nell-1 significantly increased the osteogenic gene and protein expression levels of ALP, OCN, Runx2, osteoprotegerin (OPG), collagen type I (Col-I), and Osterix (Osx) in pre-osteoblasts on Ti surfaces. The optimal concentration of Nell-1 was 100 ng/ ml. In addition, Nell-1 activated ERK and JNK, but not P38, in MC3T3-E1 cells on the Ti surface. Except for ALP and Col-I, the promotive effects of Nell-1 on the expression of osteogenic markers were suppressed by ERK inhibitor U0126. Conclusion: Certain concentrations of Nell-1 can promote the osteogenic differentiation of pre-osteoblasts on Ti surfaces by activating the MAPK/ERK signaling pathway.

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Section: Discussionmentioning

confidence: 86%

Nell-1 Enhances Osteogenic Differentiation of Pre-Osteoblasts on Titanium Surfaces via the MAPK-ERK Signaling Pathway

Shen

Wang

et al. 2018

Cell Physiol Biochem

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“…62, 66 In contrast, prior studies have suggested that assembly of NELL-1 into homo-oligomeric structures is at least partially required for its bioactive effects. 67 Fourthly, the cost of manufacturing and risk of contamination can be considerably diminished by using small molecules. 62 Unfortunately, no known small molecule has yet been developed as an agonist for NELL-1 signaling.…”

Section: Discussionmentioning

confidence: 99%

Combining Smoothened Agonist and NEL-Like Protein-1 Enhances Bone Healing

Lee

Wang

Pan

et al. 2017

Plastic &Amp; Reconstructive Surgery

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Background Non-healing bone defects represent an immense biomedical burden. Despite recent advances in protein based bone regeneration, safety concerns over bone morphogenetic protein-2 have prompted the search for alternative factors. Previously, we examined the additive/synergistic effects of Hedgehog (Hh) and Nel-like protein-1 (NELL-1) on the osteogenic differentiation of mesenchymal stem cells in vitro. Here, we sought to leverage our previous findings by applying the combination of Smoothened agonist (SAG), Hh signal activator, and NELL-1 to an in vivo critical size bone defect model. Methods A 4 mm parietal bone defect was created in mixed gender CD-1 mice. Treatment groups included control (n = 6), SAG (n = 7), NELL-1 (n = 7) or SAG + NELL-1 (n = 7). A custom fabricated poly(lactic-co-glycolic acid) disc with hydroxyapatite coating was used as an osteoinductive scaffold. Results Results at 4 and 8 weeks showed increased bone formation by micro-computed tomography analyses with either stimulus alone (SAG or NELL-1), but significantly greater bone formation with both components combined (SAG + NELL-1). This included greater bone healing scores, increased bone volume and bone thickness. Histologic analyses confirmed a significant increase in new bone formation with the combination therapy SAG + NELL-1, accompanied by increased defect vascularization. Conclusions In summary, our results suggest that combining the Hh signaling agonist SAG and NELL-1 has potential as a novel therapeutic strategy for the healing of critical-sized bone defects. Future directions will include optimization of dosage and delivery strategy for a SAG + NELL-1 combination product.

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“…Preosteoblastic cells are precursor cells of mature osteoblasts, located in bone marrow stroma in high numbers, and likely to be critically affected by wear debris during the AL process. Several studies have used MC3T3‐E1 cells as a model of preosteoblasts for differentiation and functionality of osteoblastic cells …”

Section: Discussionmentioning

confidence: 99%

“…Several studies have used MC3T3-E1 cells as a model of preosteoblasts for differentiation and functionality of osteoblastic cells. 11,20 In this study, we first established the concentration range of particulate and ion forms of Co-Cr alloy that could effectively affect the functions of preosteoblasts without severe cytotoxicity in a 72-h culture. It appeared that particles 2.5 mg mL 21 or ions 500 lM resulted in significant cell death compared to debris-free controls, while particles 0.15 mg mL 21 or ions 32 lM did not show any influence on preosteoblasts.…”

Section: Discussionmentioning

confidence: 99%

Biological responses of preosteoblasts to particulate and ion forms ofCo‐Cr alloy

Yang

Zhang

et al. 2015

J Biomedical Materials Res

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This study compared the particulate and ion forms of a cobalt-chrome (Co-Cr) alloy on the differentiation/activation of preosteoblasts. Mouse preosteoblasts (MC3T3-E1) were cultured in an osteoblast-induction medium in the presence of particulate and ion forms of a Co-Cr alloy, followed by cell proliferation and cytotoxicity evaluations. The maturation and function of osteoblasts were assessed by alkaline phosphatase (ALP) assay and related gene expressions. Both particulate and ion forms of the metals significantly reduced the proliferation of MC3T3-E1 cells in a dose-dependent manner. Similarly, cells challenged with high concentrations of particles and ions exhibited a marked cytotoxic effect and diminished expression of ALP. Real-time (RT) polymerase chain reaction (PCR) data have suggested that cells with Co-Cr particles dramatically promoted over-expression of monocyte chemo-attractant protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6), whereas Co(2+) ions treatment predominately up-regulated expressions of receptor activator of nuclear factor kappa-B ligand (RANKL), nuclear factor of activated T-cells cytoplasmic 1 (NFATc1), and down-regulated expression of osteoprotegerin (OPG) and Osterix (Osx). Overall, this study provides evidence that both Co-Cr alloy particles and metal ions interfered with the MC3T3-E1 cells for their growth, maturation, and functions. Further, Co-Cr particles exhibited stronger effects on inflammatory mediators, while metal ions showed more influence on inhibition of osteoblast differentiation and promotion of osteoclastogenesis.

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Oligomerization-induced Conformational Change in the C-terminal Region of Nel-like Molecule 1 (NELL1) Protein Is Necessary for the Efficient Mediation of Murine MC3T3-E1 Cell Adhesion and Spreading

Cited by 15 publications

References 45 publications

Nell-1 Enhances Osteogenic Differentiation of Pre-Osteoblasts on Titanium Surfaces via the MAPK-ERK Signaling Pathway

Nell-1 Enhances Osteogenic Differentiation of Pre-Osteoblasts on Titanium Surfaces via the MAPK-ERK Signaling Pathway

Combining Smoothened Agonist and NEL-Like Protein-1 Enhances Bone Healing

Biological responses of preosteoblasts to particulate and ion forms ofCo‐Cr alloy

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