scholarly journals Fish Collagen Promotes the Expression of Genes Related to Osteoblastic Activity

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mark Luigi Fabian Capati ◽  
Ayako Nakazono ◽  
Kohei Yamamoto ◽  
Kouji Sugimoto ◽  
Kajiro Yanagiguchi ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Clinical Application ◽  
Biological Effects ◽  
Pcr Analysis ◽  
Type I ◽  
Experimental Conditions ◽  
Suitable Alternative ◽  
Alkaline Phosphatase Staining ◽  
Proliferation And Differentiation

Tilapia type I atelocollagen (TAC) is a strong candidate for clinical application as its biological scaffold due to a high degeneration temperature and biologically safe properties. The aim of this study was to confirm the biological effects of TACin vitroon osteoblastic cells, simulating its clinical application. The proliferation and differentiation of typical preosteoblasts, MC3T3-E1 cells, were investigated using a microarray analysis, staining assay for mineralization, and real-time PCR analysis of the expression of mineralization-related genes. The mRNA expression of 10 genes involved in proliferation and differentiation increased after 3-day culture on an TAC gel, with an average balanced score ratio exceeding 1.5 compared to the control. After two weeks of culture, all three experimental groups showed stronger alkaline phosphatase staining than after one week. The genes expression of alkaline phosphatase, osteocalcin, and bone sialoprotein increased under the experimental conditions. The gene expression of osteopontin did not increase, and no statistical differences were noted among the three experimental groups. The present and previous findings suggest that TAC is not only a suitable alternative to collagen products originating from mammals but also a novel biomaterial with cell differentiation ability for regenerative medicine.

Effects of fluoride on human bone cells in vitro: differences in responsiveness between stromal osteoblast precursors and mature osteoblasts

1994 ◽  
Vol 130 (4) ◽  
pp. 381-386 ◽  
Author(s):  
Moustapha Kassem ◽  
Leif Mosekilde ◽  
Erik F Eriksen
Keyword(s):  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Sodium Fluoride ◽  
Bone Cells ◽  
Human Bone ◽  
Type I ◽  
Procollagen Type ◽  
Procollagen Type I ◽  
Osteoblast Precursors

Kassem M, Mosekilde L, Eriksen EF. Effects of fluoride on human bone cells in vitro: differences in responsiveness between stromal osteoblast precursors and mature osteoblasts. Eur J Endocrinol 1994;130:381–6. ISSN 0804–4643 The cellular effects of sodium fluoride (NaF) on human bone cells in vitro have been variable and dependent on the culture system used. Variability could be attributed to differences in responsiveness to NaF among different populations of cells at various stages of differentiation in the osteoblastic lineage. In this study we compared the effects of NaF in serum-free medium on cultures of more differentiated human osteoblast-like (hOB) cells derived from trabecular bone explants and on osteoblast committed precursors derived from human bone marrow, i.e. human marrow stromal osteoblast-like (hMS(OB)) cells. Sodium fluoride (10−5 mol/l) increased proliferation of hMS(OB) cells (p<0.05, N = 10) but was not mitogenic to hOB cells (p>0.05, N= 10). Alkaline phosphatase (AP) production increased in both hMS(OB) (p<0.05, N=9) and hOB cells (p<0.05, N=9). No significant effects on procollagen type I propeptide production were obtained in either culture. In the presence of 1,25-dihydroxycholecalciferol (10−9 mol/l), NaF enhanced alkaline phosphatase (p<0.05, N=8), procollagen type I propeptide (p<0.05, N=7) and osteocalcin (p<0.05, N=7) production by hMS(OB) cells but not by hOB cells. Our results suggest that osteoblast precursors are more sensitive to NaF action than mature osteoblasts and that the in vivo effects of NaF on bone formation may be mediated by stimulating proliferation and differentiation of committed osteoblast precursors in bone marrow. M Kassem, Mayo Clinic, Endocrine Research Unit, W-Joseph 5-164, Rochester, MN 55904, USA

Biocompatibility of Calcium Phosphate Ceramics Synthesized from Eggshell

2007 ◽  
Vol 330-332 ◽  
pp. 23-26 ◽  
Author(s):  
Nam Sik Oh ◽  
Yun Ho Na ◽  
S. W. Ji ◽  
S.W. Song ◽  
S.H. Oh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Alkaline Phosphatase ◽  
Cell Attachment ◽  
Morphological Characteristics ◽  
Osteoblastic Differentiation ◽  
Human Bone ◽  
Pcr Analysis ◽  
Mixing Ratio ◽  
Culture Plate ◽  
Alkaline Phosphatase Staining

The aim of this paper was the HA and β-TCP powers were synthesized by a new wetchemical method using eggshell and phosphoric acid. The biocompatibility of synthesized natural HA, HA/β-TCP(50:50) and β-TCP derived from eggshell was compared with those of as commercial chemical powder with mesenchymal stem cells derived from human bone marrow. Development of crystalline phases of the mixtures was studied as functions of mixing ratio and temperature using X-ray diffractometer. The morphological characteristics of the calcined eggshell and synthesized powders were examined by scaning electron microscopy. The in-vitro cytotoxicity and cell attachment of sintered disks were examined using human bone marrowderived multipotent stem cells(hBMSCs). Cell response was characterized by MTT assay , Alkaline phosphatase stain and RT-PCR analysis. Pure HA was synthesized in the mixing ratio of 1:1.1 wt% at 900°C for 1h. the crystallization of HA was started at 800°C in the 1:1.1 mixing ratio, ant the HA phase was continued up to the high temperatures. In the ratio of 1:1.3 and 1:1.5 wt%, β-TCP was effectively synthesized at 900°C. In the 1:1.5 ratio, β-TCP phase was detected at 700°C, and complete crystallized β-TCP was observed above 900°C. At the higher temperature than 1000°C, the β-TCP was gradually decreased and α-TCP was observed. The HA and β-TCP disk does not exert cytotoxic effect on the hBMSCs undergoing osteoblastic differentiation. In addition, the hBMSCs are adhered on the surface of synthesized natural HA and β-TCP disk as successfully as on the culture plate or as commercial chemical HA and β-TCP disk. The hBMSCs adhered on either synthesized natural HA, β-TCP or as commercial chemical HA, β-TCP disk displays undistinguishable actin arrangement and cellular phenotypes, indicating that synthesized natural HA, β-TCP does not disrupt normal cellular responses. Analysis of differentiation of the hBMSCs cultured on culture plate, synthesized natural HA, β-TCP and as commercial chemichal HA, β-TCP disk shows that three matrices are able to support osteoblastic differentiation of the hBMSCs as accessed by alkaline phosphatase staining.

scholarly journals PHAGOCYTOSIS OF BACTERIA IN THE ABSENCE OF ANTIBODY AND THE EFFECT OF PHYSICAL SURFACE

1956 ◽  
Vol 104 (2) ◽  
pp. 233-243 ◽  
Author(s):  
Edwin M. Lerner
Keyword(s):  
Filter Paper ◽  
Chemical Effect ◽  
Type I ◽  
Blood Leukocytes ◽  
Experimental Conditions ◽  
Phagocytic Capacity ◽  
E Coli ◽  
Physical Surface ◽  
Type Iv

The present experiments have shown that phagocytosis occurs in the absence of specific antibody and in the absence of a "suitable physical surface", as further that the presence of a rough surface does not increase the in vitro phagocytosis of pneumococci by polymorphonuclear leukocytes. This held true during repetition of Wood's experiments, as well as when more controlled quantitative techniques were employed, when conditions were made optimal for phagocytosis by increasing bacterial concentrations, and when blood leukocytes were substituted for exudate leukocytes. Evidence has been presented previously that the stimulation of phagocytosis of E. coli, B. abortus, and Type IV Pneumococcus, after contact with filter paper or an active compound present in filter paper, is a chemical effect rather than a physical effect. This type of stimulation did not occur with the Type I A5 Pneumococcus. The leukocyte of the circulating blood was found to be definitely superior to the exudate leukocyte in phagocytic capacity, under all the experimental conditions tested.

25 PRODUCTION OF TRANSGENIC PIGS WITH CreER-MEDIATED ASTROCYTIC-SPECIFIC RECOMBINATION SYSTEM FOR NEUROLOGICAL DISEASE MODELS

2017 ◽  
Vol 29 (1) ◽  
pp. 120
Author(s):  
S.-U. Hwang ◽  
J. D. Yoon ◽  
K. Eun ◽  
H. Kim ◽  
S.-H. Hyun
Keyword(s):  
Neurological Disease ◽  
Fluorescent Protein ◽  
Brain Magnetic Resonance Imaging ◽  
Donor Cell ◽  
Genetically Engineered ◽  
Pcr Analysis ◽  
Transgenic Pigs ◽  
Suitable Alternative ◽  
Recombination System

Pigs are one of the most suitable alternative laboratory models than other animals, because they have similar cardiovascular, renal and gastrointestinal organs with those of human. However, in the case of genetically engineered animals, early development of embryos is inhibited by expression of foreign genes, there are many cases of miscarriage or birth early mortality. To overcome these problems, we constructed pig glial fibrillary acidic protein (GFAP) promoter-Cre recombinase fused to a mutated ligand-binding domain of the human oestrogen receptor (CreERT2) and enhanced green fluorescent protein (EGFP)-LoxP transgenes for tamoxifen(TM)-inducible CreERT2-mediated recombination. We then established donor transgenic pig fibroblasts with pGFAP-CreERT2; LCMV-EGFPLoxP transgenes for somatic cell nuclear transfer (SCNT). We produced the SCNT embryos using a Cloud male #5 pGFAP-CreERT2+LCMV-EGFPLoxP donor cell line that was verified in vitro. It was transferred into a surrogate mother and then 5 pGFAP-CreERT2; EGFPLoxP TG piglets were born. By immunofluorescence staining and semi-nested PCR analysis, it was proved that CreER-mediated astrocytic-specific recombination system was operated in some cerebral astrocytic cells after TM-administration to TG pig #4. Additionally, we obtained brain magnetic resonance imaging (MRI) images using 3T-tesla MRI. Brain compartment volume (total brain, grey matter, white matter, cerebellum, brainstem, lateral ventricle, thalamus, midbrain, pons, medulla oblongata, hypophysis) was no significant differences between normal pig and pGFAP-CreERT2; EGFPLoxP transgenic (TG) pig. In summary, we verified the pGFAP promoter-driven CreERT2-LoxP recombination system in TG pig generated by SCNT depending on the TM administration. We suggest that this technology will be a useful tool for studying physiology of astrocytes and generating TG pig model of neurological disease such as Huntington’s disease, Alzheimer’s disease and brain tumour.

scholarly journals Effects of RAC1 on Proliferation of Hen Ovarian Prehierarchical Follicle Granulosa Cells

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1589
Author(s):  
Thobela Louis Tyasi ◽  
Xue Sun ◽  
Xuesong Shan ◽  
Simushi Liswaniso ◽  
Ignatius Musenge Chimbaka ◽  
...  
Keyword(s):  
Cell Cycle Progression ◽  
Biological Effects ◽  
Expression Levels ◽  
Cell Proliferation And Differentiation ◽  
Proliferation And Differentiation ◽  
Mrna And Protein Expression ◽  
Chicken Ovary ◽  
New Evidence ◽  
Follicle Selection

RAC1 belongs to the small G protein Rho subfamily and is implicated in regulating gene expression, cell proliferation and differentiation in mammals and humans; nevertheless, the function of RAC1 in growth and development of hen ovarian follicles is still unclear. This study sought to understand the biological effects of RAC1 on granulosa cell (GC) proliferation and differentiation of hen ovarian prehierarchical follicles. Firstly, our results showed expression levels of RAC1 mRNA in the follicles with diameters of 7.0–8.0 mm, 6.0–6.9 mm and 1.0–3.9 mm were greater than other follicles (p < 0.05). The RAC1 protein was mainly expressed in oocyte and its around GCs and stromal tissues of the prehierarchical follicles by immunohistochemistry. Further investigation revealed the RAC1 gene remarkably enhanced the mRNA and protein expression levels of FSHR (a marker of follicle selection), CCND2 (a marker of cell-cycle progression and GC differentiation), PCNA (a marker of GC proliferation), StAR and CYP11A1 (markers of GC differentiation and steroidogenesis) (p < 0.05). Furthermore, our data demonstrated siRNA interference of RAC1 significantly reduced GC proliferation (p < 0.05), while RAC1 gene overexpression enhanced GC proliferation in vitro (p < 0.05). Collectively, this study provided new evidence that the biological effects of RAC1 on GC proliferation, differentiation and steroidogenesis of chicken ovary follicles.

scholarly journals Isopsoralen Enhanced Osteogenesis by Targeting AhR/ERα

Molecules ◽  
2018 ◽  
Vol 23 (10) ◽  
pp. 2600 ◽  
Author(s):  
Luna Ge ◽  
Yazhou Cui ◽  
Kai Cheng ◽  
Jinxiang Han
Keyword(s):  
Osteoblast Differentiation ◽  
Estrogen Receptor Alpha ◽  
Biological Effects ◽  
Hormone Deficiency ◽  
In Vivo Studies ◽  
Osteogenic Potential ◽  
Type I ◽  
Dependent Manner

Isopsoralen (IPRN), one of the main effective ingredients in Psoralea corylifolia Linn, has a variety of biological effects, including antiosteoporotic effects. In vivo studies show that IPRN can increase bone strength and trabecular bone microstructure in a sex hormone deficiency-induced osteoporosis model. However, the mechanism underlying this osteogenic potential has not been investigated in detail. In the present study, we investigated the molecular mechanism of IPRN-induced osteogenesis in MC3T3-E1 cells. Isopsoralen promoted osteoblast differentiation and mineralization, increased calcium nodule levels and alkaline phosphatase (ALP) activity and upregulated osteoblast markers, including ALP, runt-related transcription factor 2 (RUNX2), and collagen type I alpha 1 chain (COL1A1). Furthermore, IPRN limited the nucleocytoplasmic shuttling of aryl hydrocarbon receptor (AhR) by directly binding to AhR. The AhR target gene cytochrome P450 family 1 subfamily A member 1 (CYP1A1) was also inhibited in vitro and in vivo. This effect was inhibited by the AhR agonists indole-3-carbinol (I3C) and 3-methylcholanthrene (3MC). Moreover, IPRN also increased estrogen receptor alpha (ERα) expression in an AhR-dependent manner. Taken together, these results suggest that IPRN acts as an AhR antagonist and promotes osteoblast differentiation via the AhR/ERα axis.

scholarly journals The Preventive Effect of Biochanin A on Bone Loss in Ovariectomized Rats: Involvement in Regulation of Growth and Activity of Osteoblasts and Osteoclasts

2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
Shu-Jem Su ◽  
Yao-Tsung Yeh ◽  
Huey-Wen Shyu
Keyword(s):  
Bone Loss ◽  
Mrna Expression ◽  
Biological Effects ◽  
Biochanin A ◽  
Ovariectomized Rats ◽  
Tartrate Resistant Acid Phosphatase ◽  
Type I ◽  
Mineral Density ◽  
Ovx Rats

Biochanin A (BCA) is a major isoflavone abundant in red clover (Trifolium pretense). The protective effect of BCA on bone loss in an ovariectomized (OVX) animal model has never been clarified. The objective of this study was to investigate the biological effects of BCA on bone loss in OVX ratsin vivoand on the development of osteoblasts and osteoclastsin vitro. Ovariectomy resulted in a marked increase in body weight and a decrease in femoral bone mineral density and trabecular bone volume that was prevented by BCA or 17β-estradiol (E2) treatment. However, an increase in uterine weight was observed in E2-treated OVX rats, but not in response to BCA treatment. Treatment with BCA increased the mRNA expression of osterix, collagen type I, alkaline phosphatase (ALP), and osteocalcin and decreased the mRNA expression of tartrate-resistant acid phosphatase (TRAP) and the receptor activator of nuclear factor-κB ligand (RANKL)/osteoprotegerin (OPG) ratio in the femur of OVX rats. Treatment with BCA or E2 prevented the OVX-induced increase in urinary deoxypyridinoline (DPD) and serum tumor necrosis factorα(TNF-α) and interleukin-1β(IL-1β).In vitro, BCA induced preosteoblasts to differentiate into osteoblasts and increased osteoblast mineralization. BCA inhibited preosteoclasts and osteoclast proliferation and decreased osteoclast bone resorption. These findings suggest that BCA treatment can effectively prevent the OVX-induced increase in bone loss and bone turnover possibly by increasing osteoblastic activities and decreasing osteoclastic activities.

scholarly journals Expression and homologous regulation of GH secretagogue receptor mRNA in rat adrenal gland

2002 ◽  
pp. 677-688 ◽  
Author(s):  
ML Barreiro ◽  
L Pinilla ◽  
E Aguilar ◽  
M Tena-Sempere
Keyword(s):  
Biological Effects ◽  
Biologically Active ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
R Gene ◽  
Rt Pcr ◽  
Short Term ◽  
Opposite Pattern ◽  
Corticosterone Secretion

OBJECTIVE: GH secretagogues (GHSs) elicit a variety of biological effects in several endocrine and non-endocrine target tIssues, including activation of the hypothalamic-pituitary-adrenal axis. The latter is mainly carried out through a central hypothalamic action; yet the possibility of additional effects directly at the adrenal level cannot be ruled out. The aims of this study were to evaluate the expression and homologous regulation of the GHS-receptor (GHS-R) gene in rat adrenal and to assess the effects of synthetic (GH releasing peptide-6 - GHRP-6) and natural (ghrelin) ligands of GHS-R upon basal and ACTH-stimulated corticosterone secretion in vitro. DESIGN AND METHODS: Analysis of adrenal expression of target mRNAs (GHS-R, GHS-R1a, ghrelin, and several steroidogenic factors) was conducted by means of primer-specific, semi-quantitative RT-PCR. Evaluation of corticosterone secretion by incubated adrenal tIssue was carried out by specific RIA. RESULTS: RT-PCR analysis demonstrated expression of the GHS-R gene, but not of the gene encoding the cognate ligand ghrelin, in rat adrenal. Moreover, expression of the mRNA coding for the type 1a GHS-R (GHS-R1a), i.e. the biologically active receptor form, was demonstrated. The adrenal expression of the GHS-R message appeared under the regulation of homologous signals in vitro, as short-term incubation of adrenal samples in serum-free medium induced a significant increase in GHS-R mRNA levels that was inhibited by exposure to different doses of GHRP-6 (10(-9)-10(-5) mol/l) or ghrelin (10(-7) mol/l). Notably, an opposite pattern of homologous regulation of GHS-R gene expression was observed at the pituitary. Finally, short-term stimulation with increasing concentrations of GHRP-6 (10(-9)-10(-5) mol/l) or ghrelin (10(-7) mol/l) failed to alter basal and ACTH-stimulated corticosterone secretion in vitro, neither did it modify ACTH-stimulated mRNA expression levels of several upstream elements in the steroidogenic route: the steroidogenic acute regulatory (StAR) protein, and the enzymes P450 cholesterol side-chain cleavage (P450scc) and 3beta-hydroxysteroid dehydrogenase (3beta-HSD). CONCLUSIONS: Our study provides novel evidence for the expression and homologous regulation of the GHS-R gene in rat adrenal. However, our results cast doubts on the possibility of direct adrenal actions of ligands of the GHS-R in the regulation of corticosterone secretion in the rat.

The Effect of 3D Construction Culture of Human Chondrocytes Using Alginate Sponge

2006 ◽  
Vol 326-328 ◽  
pp. 883-888 ◽  
Author(s):  
Jin Sang Lee ◽  
Byung Kim ◽  
Min Soo Kim ◽  
Seung Jae Lee ◽  
Sung Won Kim ◽  
...  
Keyword(s):  
Cartilage Tissue Engineering ◽  
Collagen Type I ◽  
Cartilage Tissue ◽  
Type I ◽  
In Vitro Production ◽  
3D Scaffold ◽  
Proliferation And Differentiation ◽  
Electron Microscopy Analysis ◽  
Vitro Production

In this study, we investigated the effect of the use of alginate sponge as a chondrocyte-3D scaffold for the construction of a cartilage graft. Alginate sponge was made by 5% alginic acid which was crosslinked by CaCl2. Chondrocytes were obtained from a nasal septum after the operation and cultured in 3D alginate sponge. For analysis of cell differentiation, we have checked aggrecan, collagen type I and II using RT-PCR and performed the histological and scanning electron microscopy analysis. Our experiments showed that alginate sponge of 5% promoted sufficient chondrocyte proliferation and differentiation, resulting in the formation of a specific cartilage matrix. The sponge presents new perspectives with respect to in vitro production of "artificial" cartilage. We conclude that the alginate sponges have potential as a scaffold for cartilage tissue engineering.

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