Stimulation of interleukin-4 of cell proliferation and mRNA expression of alkaline phosphatase and collagen type I in human osteoblast-like cells of trabecular bone

Nohtomi, Kyoko; Sato, Kanji; Shizume, Kazuo; Yamazaki, Kazuko; Demura, Hiroshi; Hosoda, Kei; Murata, Yoshiharu; Seo, Hisao

doi:10.1016/s0169-6009(08)80188-2

Cited by 20 publications

(10 citation statements)

References 23 publications

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“…As expected, thyroid glands of control rats expressed mRNA of VEGF receptors, flt-I and its related gene KDR/flk-i. Interestingly, both flt-i and KDR/flk-I mRNAs were also increased on days 3-7, and decreased gradually to the control level on days [14][15][16][17][18][19][20][21][22][23][24][25][26][27][28] (Fig. 12, B and C).…”

Section: Resultsmentioning

confidence: 99%

“…Usually, 10-cm dishes were pretreated with agarose to culture thyroid follicles in suspension. After appropriate culture periods, the thyroid follicles were centrifuged and total RNA was extracted as reported previously (18).…”

Section: Methodsmentioning

confidence: 99%

“…RNA samples were transferred onto nylon membranes and then hybridized with human VEGF cDNA spanning nucleotide 0.6 kb (6,20,21), rat VEGF 0.5-kb cDNA, rat flt-i 1.2-kb cDNA, or rat KDR/flk-J 1.2-kb cDNA (21) SDS at 550C and exposed to X-omat film (Eastman Kodak, Rochester, NY) with an intensifying screen at -700C. For comparison of RNA loading, the filters were rehybridized with 32P-labeled human /3-actin or glyceraldehyde 3-phosphate dehydrogenase (GAPDH) cDNA (supplied by Y. Murata, Research Institute for Environmental Medicine, Nagoya University, Nagoya, Japan) (22), as described previously (18).…”

Section: Methodsmentioning

confidence: 99%

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Stimulation by thyroid-stimulating hormone and Grave's immunoglobulin G of vascular endothelial growth factor mRNA expression in human thyroid follicles in vitro and flt mRNA expression in the rat thyroid in vivo.

Sato¹,

Yamazaki²,

Shizume³

et al. 1995

J. Clin. Invest.

160

113

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Stimulation by thyroid-stimulating hormone and Grave's immunoglobulin G of vascular endothelial growth factor mRNA expression in human thyroid follicles in vitro and flt mRNA expression in the rat thyroid in vivo.

Sato¹,

Yamazaki²,

Shizume³

et al. 1995

J. Clin. Invest.

160

113

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show abstract

“…T helper lymphocytes (Th2 cells) secrete IL-4, -5, -10, and -13, which are generally anti-inflammatory by inhibiting the activity of cytokines secreted from Th1 cells, such as IL-1, -2, -6, and -12 [35]. IL-4 stimulates proliferation and mineralization by enhancing hydroxyproline accumulation, calcium deposition, and osteocalcin secretion [36], [37]; because we observed attenuated response to IL-4 and osteogenic differentiation in AnxA2 kd and AnxA5 kd cells, it is possible that AnxA2 and AnxA5 function as regulators of transcription in osteoblastic cells. Indeed, Belluoccio et al reported significant alteration in the transcriptome of compound AnxA5 −/− ;AnxA6 −/− , wherein 56% of differentially-regulated genes (defined by 3-fold difference compared to control) were related to cell survival, involving cell motility, apoptosis, cell cycle, cell proliferation and differentiation and tumor suppressor/survival [8].…”

Section: Discussionmentioning

confidence: 99%

Impaired Osteoblast Differentiation in Annexin A2- and -A5-Deficient Cells

et al. 2014

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Annexins are a class of calcium-binding proteins with diverse functions in the regulation of lipid rafts, inflammation, fibrinolysis, transcriptional programming and ion transport. Within bone, they are well-characterized as components of mineralizing matrix vesicles, although little else is known as to their function during osteogenesis. We employed shRNA to generate annexin A2 (AnxA2)- or annexin A5 (AnxA5)-knockdown pre-osteoblasts, and determined whether proliferation or osteogenic differentiation was altered in knockdown cells, compared to pSiren (Si) controls. We report that DNA content, a marker of proliferation, was significantly reduced in both AnxA2 and AnxA5 knockdown cells. Alkaline phosphatase expression and activity were also suppressed in AnxA2- or AnxA5-knockdown after 14 days of culture. The pattern of osteogenic gene expression was altered in knockdown cells, with Col1a1 expressed more rapidly in knock-down cells, compared to pSiren. In contrast, Runx2, Ibsp, and Bglap all revealed decreased expression after 14 days of culture. In both AnxA2- and AnxA5-knockdown, interleukin-induced STAT6 signaling was markedly attenuated compared to pSiren controls. These data suggest that AnxA2 and AnxA5 can influence bone formation via regulation of osteoprogenitor proliferation, differentiation, and responsiveness to cytokines in addition to their well-studied function in matrix vesicles.

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“…Other local cells, especially macrophages and T cells will also contribute to the presence of various cytokines and growth factors. For instance, interleukin‐4, produced by activated T cells has been documented to stimulate not only the proliferation of osteoblast‐like cells, but also the expression of ALP mRNA and ALP enzyme activity (Nohtomi et al 1994). Thus, capillary proliferation into the wound area is enhanced and osteoblast precursor cells originating from perivascular loose connective tissue seem to be attracted to the area.…”

Section: Discussionmentioning

confidence: 99%

Temporal and local appearance of alkaline phosphatase activity in early stages of guided bone regeneration

Stucki¹,

Schmid

Hämmerle

et al. 2001

Clinical Oral Implants Res

121

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Alkaline phosphatase (ALP) catalyzes the hydrolysis of phosphate esters and it seems to be a prerequisite for normal skeletal mineralization. Also, ALP is the most widely recognized marker of osteoblast phenotypes. By a tissue regenerative technique called Guided Bone Regeneration (GBR), it is possible nowadays to regenerate small bony defects. The aim of the present study was to investigate early events in bone healing and neogenesis by studying histochemically the temporal and local appearance of the marker Alkaline Phosphatase (ALP) in a GBR model system. Nine healthy volunteers (5 males, 4 females, mean age 31.7 years) participated in the experiment. After raising a mucoperiosteal flap from the mandibular second molar to the retromolar area in each volunteer, a hollow titanium test cylinder was placed into a congruent bony bed and the coronal end of the cylinder was closed with an ePTFE-membrane. Then the flap was adapted and sutured to obtain primary wound closure. After 2, 7 and 12 weeks, the regenerated tissue within the cylinders was harvested. Histologically, ALP activity was observed associated with the osteoid seams in the very basal part of the regenerate where new bone trabeculae were in the process of being formed. More coronally, large round cells seemed to secrete an ALP-positive substance since in the center of such cell clusters strong ALP activity located extracellularly was detected. In the present experiment, ALP seemed to have been an early sign of osteoblast secretion of a matrix which subsequently was determined to become osteoid. ALP activity was never seen isolated within connective tissue and away from bone. This is an indication that its source is linked to existing bone. The present study has documented for the first time the appearance of ALP activity in guided bone regenerations in humans. It has revealed that: 1) Osteogenesis in guided bone regeneration is preceded by localized, marked expression of ALP in an organized connective tissue environment. 2) Bone neogenesis is an early event in this experimental setup and may be detected already 2 weeks after wounding. 3) Expression of ALP and subsequent bone neogenesis is originating from and topographically linked to pre-existing bone structures.

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Stimulation of interleukin-4 of cell proliferation and mRNA expression of alkaline phosphatase and collagen type I in human osteoblast-like cells of trabecular bone

Cited by 20 publications

References 23 publications

Stimulation by thyroid-stimulating hormone and Grave's immunoglobulin G of vascular endothelial growth factor mRNA expression in human thyroid follicles in vitro and flt mRNA expression in the rat thyroid in vivo.

Stimulation by thyroid-stimulating hormone and Grave's immunoglobulin G of vascular endothelial growth factor mRNA expression in human thyroid follicles in vitro and flt mRNA expression in the rat thyroid in vivo.

Impaired Osteoblast Differentiation in Annexin A2- and -A5-Deficient Cells

Temporal and local appearance of alkaline phosphatase activity in early stages of guided bone regeneration

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