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.

Impaired Osteogenic Differentiation Of Mesenchymal Stem Cells Derived From Bone Marrow Of Patients With Low-Risk Myelodysplastic Syndromes

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 1579-1579
Author(s):  
Chunkang Chang ◽  
Chengming Fei ◽  
Youshan Zhao ◽  
Juan Guo ◽  
Xiao Li
Keyword(s):  
Alkaline Phosphatase ◽  
Osteogenic Differentiation ◽  
Lower Risk ◽  
Conflicts Of Interest ◽  
Osteoblastic Differentiation ◽  
Differentiation Markers ◽  
Alizarin Red ◽  
Hematopoietic Microenvironment ◽  
Alkaline Phosphatase Staining ◽  
Osteogenic Induction

Abstract Background The pathogenesis of MDS has not been completely understood, and insufficiency of the hematopoietic microenvironment can be an important factor. MSCs and osteoblasts are key components of the hematopoietic microenvironment. Studying osteoblastic differentiation of MSCs quantitatively may help to understand the pathogenesis of MDS. Methods 38 patients with MDS and 15 normal donors were investigated in this study. Osteoblastic differentiation assays were performed in 16 MDS cases and 8 controls. The expression of osteogenic differentiation markers were measured by real-time PCR. Alkaline phosphatase staining was performed with Alkaline Phosphatase staining kit after 3,7,14 days of incubation. ALP activity was assessed at 3, 7, and 10 days after osteogenic differentiation. Mineralization analysis was performed at 7, 14 and 21 days of osteogenic induction. The areas of mineralization were measured by Image-Pro Plus 6.0 software. Results Both MDS-MSCs and normal cells displayed same fibroblast-like morphology and similar antigen expression. The expression level of RUNX2 was significantly decreased in MSCs from MDS, compaired with normal controls, especially in lower-risk MDS. After osteogenic induction, lower-risk MDS showed lower alkaline phosphatase activity, less intense alizarin red S staining, and lower gene expression of osteogenic differentiation markers, however, higher-risk MDS was normal. Conclusions We concluded that impaired osteogenic differentiation of MSCs was seen mainly in patients with lower-risk MDS. It may contribute to the ineffective hamatopoiesis of MDS. Disclosures: No relevant conflicts of interest to declare.

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.

Stimulatory Effect of Nano-Sized Calcium Metaphosphate Particles on Proliferation and Osteoblastic Differentiation of Human Bone Marrow Mesenchymal Stem Cells

2007 ◽  
Vol 361-363 ◽  
pp. 1177-1180 ◽  
Author(s):  
Sun Young Lee ◽  
Min Jung Son ◽  
Gil Son Khang ◽  
Young Suk Son ◽  
Chang Kuk You ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Morphological Changes ◽  
Osteoblastic Differentiation ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Marrow Mesenchymal Stem Cells ◽  
Potential Applications ◽  
Biomedical Field ◽  
Dose Dependent

Recently, nanomaterials have received considerable attention because of their potential applications in the biomedical field. In the present study, we investigated the effects of nano-sized calcium metaphosphate (CMP) particles (50 nm) compared with micro-sized CMP particles (200-500 nm and 10 μm) on the proliferation and osteoblastic differentiation of human bone marrow stem cells (BMSCs). BMSCs were challenged with CMP particles with different sizes for 3, 5, and 7 days. An analysis of the proliferation revealed that the nano-sized CMP particles (50 nm) stimulated the proliferation of BMSCs up to 27.79% compared to the untreated control. This stimulatory effect of the nano-sized CMP particle was dose-dependent. CMP particles appeared to adhere on the surface of BMSCs but this did not cause distinguishable morphological changes. Moreover, all CMP particles (50 nm to 10 μm) were capable of stimulating an osteoblastic differentiation of BMSCs as accessed by alkaline phosphatase (ALP) and von Kossa stainings. Further molecular analysis revealed that all the CMP particles induced an expression of osteoblast-related genes such as osteocalcin (OC) and collagen I (Col I). Taken together, our data demonstrate that nano-sized CMP particles have the potential to stimulate the proliferation and osteoblastic differentiation of BMSCs.

In Vitro Biocompatibility Testing of Collagen-Calcium Phosphate Composites Using Human Bone Derived Osteoblasts

1998 ◽  
Vol 550 ◽  
Author(s):  
A.C. Lawson ◽  
M. Oyama ◽  
M.E. Emerton ◽  
M.J.O. Francis ◽  
A.H.R.W. Simpson ◽  
...  
Keyword(s):  
Alkaline Phosphatase ◽  
Calcium Phosphate ◽  
Phosphatase Activity ◽  
Alkaline Phosphatase Activity ◽  
Transforming Growth Factor ◽  
Cell Attachment ◽  
Octacalcium Phosphate ◽  
Histochemical Staining ◽  
Human Bone

AbstractHuman bone derived osteoblasts were cultured on collagen-calcium phosphate composites. The ability of the substrates to support cell attachment, proliferation and bone formation was assessed using histochemical staining for alkaline phosphatase activity and immunolocalisation of transforming growth factor- β1and type 1 collagen. The effect of calcium phosphate phase and crystal size was investigated and the calcified samples compared with uncalcified collagen. Osteoblasts adhere to the collagen-calcium phosphate composites and express a mature osteoblast phenotype in vitro. Cell adhesion was greater on unmineralised collagen than on the mineralised composites, however, these cells were less differentiated. The presence of larger crystals seemed to have a detrimental effect on the cells, reducing proliferation and alkaline phosphatase activity. There was no discernible difference between the effect of hydroxyapatite and octacalcium phosphate on the cells.

scholarly journals Human Amnion-Derived Mesenchymal Stem Cells Promote Osteogenic Differentiation in Human Bone Marrow Mesenchymal Stem Cells by Influencing the ERK1/2 Signaling Pathway

2016 ◽  
Vol 2016 ◽  
pp. 1-12 ◽  
Author(s):  
Yuli Wang ◽  
Fei Jiang ◽  
Yi Liang ◽  
Ming Shen ◽  
Ning Chen
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Alkaline Phosphatase ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Osteoblastic Differentiation ◽  
Human Amnion ◽  
Bone Deficiency ◽  
Marrow Mesenchymal Stem Cells

Human amnion-derived mesenchymal stem cells (HAMSCs) are considered to be an important resource in the field of tissue engineering because of their anti-inflammatory properties and fewer ethical issues associated with their use compared with other sources of stem cells. HAMSCs can be obtained from human amniotic membranes, a readily available and abundant tissue. However, the potential of HAMSCs as seed cells for treating bone deficiency is unknown. In this study, HAMSCs were used to promote proliferation and osteoblastic differentiation in human bone marrow mesenchymal stem cells (HBMSCs) in a Transwell coculture system. Proliferation levels were investigated by flow cytometry and immunofluorescence staining of 5-ethynyl-2′-deoxyuridine (EdU). Osteoblastic differentiation and mineralization were evaluated in chromogenic alkaline phosphatase (ALP) activity substrate assays, Alizarin red S staining, and RT-PCR analysis of early HBMSCs osteogenic marker expression. We demonstrated that HAMSCs stimulated increased alkaline phosphatase (ALP) activity, mRNA expression of osteogenic marker genes, and mineralized matrix deposition. Moreover, the effect of HAMSCs was significantly inhibited by U0126, a highly selective inhibitor of extracellular signaling-regulated kinase 1/2 (ERK1/2) signaling. We demonstrate that HAMSCs promote osteogenic differentiation in HBMSCs by influencing the ERK1/2 signaling pathway. These observations confirm the potential of HAMSCs as a seed cell for the treatment of bone deficiency.

scholarly journals Xenogenic bone matrix extracts induce osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells

2007 ◽  
Vol 2 (4) ◽  
pp. 383-390 ◽  
Author(s):  
Marwan E El-Sabban ◽  
Hilda El-Khoury ◽  
Rima Hamdan-Khalil ◽  
Steen Sindet-Pedersen ◽  
Ali Bazarbachi
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Bone Matrix ◽  
Osteoblastic Differentiation ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Xenogenic Bone

scholarly journals Differentiation of Urine-Derived Induced Pluripotent Stem Cells to Neuron, Astrocyte and Microvascular Endothelial Cells from a Diabetic Patient

2019 ◽  
Author(s):  
Wan Liu ◽  
Ping Zhang ◽  
Jing Tan ◽  
Yongzhong Lin
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Induced Pluripotent Stem Cells ◽  
Pluripotent Stem Cells ◽  
Disease Model ◽  
Morphological Characteristics ◽  
Microvascular Endothelial Cells ◽  
Alkaline Phosphatase Staining ◽  
Microvascular Endothelial ◽  
Induced Pluripotent

AbstractBackgroundComplications of central nervous system (CNS) in type 2 diabetes mellitus (T2DM) often lead to cognitive impairment and seriously affect the quality of life. However, there is no individualized disease model. Urinary epithelial cells (UECs) can be an ideal source for generating human induced pluripotent stem cells (hiPSCs) and progenitors, as they are easily accessible, non-invasive and universally available. Therefore, we intended to differentiate urine-derived hiPSCs into neuron (N), astrocyte (A) and microvascular endothelial cells (E) from a T2DM patient for future study its pathogenesis and precision medical treatment.Methods and ResultshiPSCs was successfully induced from UECs using integration free Sendai virus technology in a totally noninvasive manner. It had a normal karyotype (46, XY) and were proved to be pluripotent by immunofluorescence staining, alkaline phosphatase staining, karyotyping, teratoma experiments and methylated analysis. N, A and E were successfully induced and displayed typical morphological characteristics.ConclusionsThis study indicates that N, A, E can be generated from urine-derived hiPSCs. Then we intend to create a new disease model in vitro to simulate the cerebral microenvironment of DM which will provide new methods for further investigate the disease-specific mechanisms.

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