Aesculetin Accelerates Osteoblast Differentiation and Matrix-Vesicle-Mediated Mineralization

The imbalance between bone resorption and bone formation in favor of resorption results in bone loss and deterioration of bone architecture. Osteoblast differentiation is a sequential event accompanying biogenesis of matrix vesicles and mineralization of collagen matrix with hydroxyapatite crystals. Considerable efforts have been made in developing naturally-occurring plant compounds, preventing bone pathologies, or enhancing bone regeneration. Coumarin aesculetin inhibits osteoporosis through hampering the ruffled border formation of mature osteoclasts. However, little is known regarding the effects of aesculetin on the impairment of matrix vesicle biogenesis. MC3T3-E1 cells were cultured in differentiation media with 1–10 μM aesculetin for up to 21 days. Aesculetin boosted the bone morphogenetic protein-2 expression, and alkaline phosphatase activation of differentiating MC3T3-E1 cells. The presence of aesculetin strengthened the expression of collagen type 1 and osteoprotegerin and transcription of Runt-related transcription factor 2 in differentiating osteoblasts for 9 days. When ≥1–5 μM aesculetin was added to differentiating cells for 15–18 days, the induction of non-collagenous proteins of bone sialoprotein II, osteopontin, osteocalcin, and osteonectin was markedly enhanced, facilitating the formation of hydroxyapatite crystals and mineralized collagen matrix. The induction of annexin V and PHOSPHO 1 was further augmented in ≥5 μM aesculetin-treated differentiating osteoblasts for 21 days. In addition, the levels of tissue-nonspecific alkaline phosphatase and collagen type 1 were further enhanced within the extracellular space and on matrix vesicles of mature osteoblasts treated with aesculetin, indicating matrix vesicle-mediated bone mineralization. Finally, aesculetin markedly accelerated the production of thrombospondin-1 and tenascin C in mature osteoblasts, leading to their adhesion to preformed collagen matrix. Therefore, aesculetin enhanced osteoblast differentiation, and matrix vesicle biogenesis and mineralization. These findings suggest that aesculetin may be a potential osteo-inductive agent preventing bone pathologies or enhancing bone regeneration.

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Direct comparison of 3D and 2D cultivation reveals higher osteogenic capacity of elderly osteoblasts in 3D

Journal of Orthopaedic Surgery and Research ◽

10.1186/s13018-020-02153-z ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Stephan Payr ◽

Elizabeth Rosado-Balmayor ◽

Thomas Tiefenboeck ◽

Tim Schuseil ◽

Marina Unger ◽

...

Keyword(s):

Gene Expression ◽

Collagen Type ◽

3D Culture ◽

Osteogenic Potential ◽

Donor Age ◽

Collagen Type 1 ◽

2D And 3D ◽

Human Primary Osteoblasts ◽

Gene Expression Levels

Abstract Background The aim of this study was the investigation of the osteogenic potential of human osteoblasts of advanced donor age in 2D and 3D culture. Methods Osteoblasts were induced to osteogenic differentiation and cultivated, using the same polystyrene material in 2D and 3D culture for 2 weeks. Samples were taken to evaluate alkaline phosphatase (ALP) activity, mineralization and gene expression. Results Osteoprotegerin (OPG) levels were significantly increased (8.2-fold) on day 7 in 3D compared to day 0 (p < 0.0001) and 11.6-fold higher in 3D than in 2D (p < 0.0001). Both culture systems showed reduced osteocalcin (OC) levels (2D 85% and 3D 50% of basic value). Collagen type 1 (Col1) expression was elevated in 3D on day 7 (1.4-fold; p = 0.009). Osteopontin (OP) expression showed 6.5-fold higher levels on day 7 (p = 0.002) in 3D than in 2D. Mineralization was significantly higher in 3D on day 14 (p = 0.0002). Conclusion Advanced donor age human primary osteoblasts reveal significantly higher gene expression levels of OPG, Col1 and OP in 3D than in monolayer. Therefore, it seems that a relatively high potential of bone formation in a natural 3D arrangement is presumably still present in osteoblasts of elderly people. Trial registration 5217/11 on the 22nd of Dec. 2011.

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Evaluation of Collagen Type-1 Production and Anti-Inflammatory Activities of Human Placental Extracts in Human Gingival Fibroblasts

Journal of Hard Tissue Biology ◽

10.2485/jhtb.25.277 ◽

2016 ◽

Vol 25 (3) ◽

pp. 277-281 ◽

Cited By ~ 1

Author(s):

Homare Akagi ◽

Yasuhiro Imamura ◽

Yoshimasa Makita ◽

Hiroe Nakamura ◽

Naomi Hasegawa ◽

...

Keyword(s):

Collagen Type ◽

Human Gingival Fibroblasts ◽

Gingival Fibroblasts ◽

Anti Inflammatory ◽

Collagen Type 1

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The Effect of Ozone on Collagen Type-1 and Inflammatory Cytokine Production in Human Gingival Fibroblasts

Dentistry ◽

10.4172/2161-1122.1000339 ◽

2015 ◽

Vol 5 (10) ◽

Cited By ~ 1

Author(s):

Yoshimasa Makita Yasuhiro Imamura ◽

Kazuya Masuno Isao Tamura

Keyword(s):

Inflammatory Cytokine ◽

Collagen Type ◽

Cytokine Production ◽

Human Gingival Fibroblasts ◽

Gingival Fibroblasts ◽

Inflammatory Cytokine Production ◽

Collagen Type 1

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Novel biomimetic tripolymer scaffolds consisting of chitosan, collagen type 1, and hyaluronic acid for bone marrow-derived human mesenchymal stem cells-based bone tissue engineering

Journal of Biomedical Materials Research Part B Applied Biomaterials ◽

10.1002/jbm.b.33152 ◽

2014 ◽

Vol 102 (8) ◽

pp. 1825-1834 ◽

Cited By ~ 19

Author(s):

Smitha Mathews ◽

Ramesh Bhonde ◽

Pawan Kumar Gupta ◽

Satish Totey

Keyword(s):

Tissue Engineering ◽

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Hyaluronic Acid ◽

Bone Tissue Engineering ◽

Collagen Type ◽

Human Mesenchymal Stem Cells ◽

Collagen Type 1

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Evaluation of Secretome Tenogenic Potential from Adipose Stem Cells (ACS) in Hypoxic Condition with Fresh Frozen Tendon Scaffold Using Scleraxis (Scx), Insulin-Like Growth Factor 1 (IGF-1) and Collagen Type 1

Journal of Biomimetics Biomaterials and Biomedical Engineering ◽

10.4028/www.scientific.net/jbbbe.49.111 ◽

2021 ◽

Vol 49 ◽

pp. 111-118

Author(s):

Ramadhan Ananditia Putra ◽

Heri Suroto

Keyword(s):

Cell Culture ◽

Collagen Type ◽

Hypoxic Condition ◽

Adipose Stem Cells ◽

Immunohistochemical Examination ◽

Hypoxic Conditions ◽

Fresh Frozen ◽

Collagen Type 1

Various studies have been conducted to see the scaffold that supports the regeneration of tendon. This study aims to analyze thein vitrosecretome tenogenic potential produced by ASCs culture with fresh frozen tendon scaffold in hypoxic conditions. ELISA tests for Scx and IGF-1 levels in secretome were obtained from ASC culture with fresh frozen tendon scaffold under normoxic (21%) and hypoxia (2%) conditions. The immunohistochemical examination of COL-1 was also carried out on the 2ndand 6thdays of cell culture. The secretion of Scx and IGF-1 was increased in secretome from ASC cultures using a fresh frozen tendon scaffold compared with those which did not (p <0.05). In the normoxia condition, Scx and IGF-1 in secretome with fresh frozen tendons had better results than hypoxic conditions (p <0.05). The highest Scx levels were obtained in culture on the 6thday (p <0.05), while the highest IGF-1 levels were obtained in the culture on the 2ndday (p <0.05). There was an increase in the secretion of Scx and IGF-1 from ASC cultures with fresh frozen tendon scaffold under the hypoxic condition of 2%.

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1,25-Dihydroxyvitamin D3 Reduces TGF-β3-Induced Fibrosis-Related Gene Expression in Human Uterine Leiomyoma Cells

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2010-2131 ◽

2011 ◽

Vol 96 (4) ◽

pp. E754-E762 ◽

Cited By ~ 83

Author(s):

Sunil K. Halder ◽

J. Shawn Goodwin ◽

Ayman Al-Hendy

Keyword(s):

Protein Expression ◽

Vitamin D3 ◽

Uterine Leiomyoma ◽

Collagen Type ◽

Plasminogen Activator Inhibitor ◽

Plasminogen Activator Inhibitor 1 ◽

Dihydroxyvitamin D3 ◽

Collagen Type 1 ◽

Activator Inhibitor

Abstract Background: Uterine leiomyomas (fibroids) are the most common benign estrogen-dependent tumors of premenopausal women. TGF-β3 up-regulates the synthesis of many of extracellular matrix proteins that are associated with tissue fibrosis. Objective: To examine the effect of 1,25-dihydroxyvitamin D3 (vitamin D3) on TGF-β3-induced fibrosis-related protein expression in immortalized human uterine leiomyoma (HuLM) cells. Methods: HuLM cells were treated with TGF-β3 with or without vitamin D3. Western blot analyses were employed to test the effect of vitamin D3 on TGF-β3-induced protein expression of collagen type 1, fibronectin, and plasminogen activator inhibitor-1 proteins. Western blots as well as immunofluorescence analyses were used to verify the effect of vitamin D3 on TGF-β3-induced Smad activation involved in extracellular matrix protein synthesis and deposition, which ultimately lead to tissue fibrosis. Results: We observed that TGF-β3 induced fibronectin and collagen type 1 protein expression in HuLM cells, and that effect was suppressed by vitamin D3. TGF-β3 also induced protein expression of plasminogen activator inhibitor-1, an important TGF-β target, in HuLM cells, which was also inhibited by vitamin D3. Additionally, TGF-β3 induced phosphorylation of Smad2 as well as nuclear translocation of Smad2 and Smad3 in HuLM cells, whereas vitamin D significantly reduced all these TGF-β3-mediated effects. Therefore, our results suggest that vitamin D3 has consistently reduced TGF-β3 effects that are involved in the process of fibrosis in human leiomyoma cells. Conclusion: Vitamin D3 is an antifibrotic factor that might be potentially useful as a novel therapeutic for nonsurgical treatment of benign uterine fibroids.

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β1-Integrin binding to collagen type 1 transmits breast cancer cells into chemoresistance by activating ABC efflux transporters

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research ◽

10.1016/j.bbamcr.2020.118663 ◽

2020 ◽

Vol 1867 (5) ◽

pp. 118663 ◽

Cited By ~ 7

Author(s):

Fabian Baltes ◽

Vladlena Pfeifer ◽

Katja Silbermann ◽

Julia Caspers ◽

Kathleen Wantoch von Rekowski ◽

...

Keyword(s):

Breast Cancer ◽

Cancer Cells ◽

Breast Cancer Cells ◽

Collagen Type ◽

Efflux Transporters ◽

Β1 Integrin ◽

Collagen Type 1

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Cystathionine-β-synthase gene transfer and 3-deazaadenosine ameliorate inflammatory response in endothelial cells

AJP Cell Physiology ◽

10.1152/ajpcell.00207.2007 ◽

2007 ◽

Vol 293 (6) ◽

pp. C1779-C1787 ◽

Cited By ~ 30

Author(s):

Utpal Sen ◽

Neetu Tyagi ◽

Munish Kumar ◽

Karni S. Moshal ◽

Walter E. Rodriguez ◽

...

Keyword(s):

Endothelial Cells ◽

Inflammatory Response ◽

Protein Expression ◽

Adhesion Molecules ◽

Adhesion Molecule ◽

Vascular Remodeling ◽

Collagen Type ◽

Gene Transfection ◽

Collagen Type 1

Although elevated levels of homocysteine (Hcy) known as hyperhomocysteinemia (HHcy) are associated with increased inflammation and vascular remodeling, the mechanism of Hcy-mediated inflammation and vascular remodeling is unclear. The matrix metalloproteinases (MMPs) and adhesion molecules play an important role in vascular remodeling. We hypothesized that HHcy induces inflammation by increasing adhesion molecules and matrix protein expression. Endothelial cells were supplemented with high methionine, and Hcy accumulation was measured by HPLC. Nitric oxide (NO) bioavailability was detected by a NO probe. The protein expression was measured by Western blot analysis. MMP-9 activity was detected by gelatin-gel zymography. We demonstrated that methionine supplement promoted upregulation of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) through increased Hcy accumulation. In addition, increased synthesis of collagen type-1 was also observed. MMP-9 gene expression and protein activity were increased in methionine supplement groups. 3-Deazaadenosine (DZA), an adenosine analogue, prevented high methionine-induced ICAM-1 and VCAM-1 expression and collagen type-1 synthesis. Transfection of endothelial cells with cystathionine-β-synthase (CBS) gene construct, which converts Hcy to cystathionine, reduced Hcy accumulation in high methionine-fed cells. CBS gene transfection reduced the inflammatory response, as evident by attenuated ICAM-1 and VCAM-1 expression. Furthermore, collagen type-1 expression and MMP-9 activity were dramatically attenuated with CBS gene transfection. These results suggested that methionine supplement increased Hcy accumulation, which was associated with inflammatory response and matrix remodeling such as collagen type-1 synthesis and MMP-9 activity. However, in vitro DZA and CBS gene therapy successfully treated the HHcy-induced inflammatory reaction in the methionine metabolism pathway.

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