Bone Regeneration Using Rat-Derived Dedifferentiated Fat Cells Combined with Activated Platelet-Rich Plasma

Bone regeneration using mesenchymal stem cells has several limitations. We investigated adipose-derived dedifferentiated fat (DFAT) cells as an alternative, and evaluated their cell proliferation rate, osteoblast differentiation, and bone regeneration ability in combination with activated platelet-rich plasma (aPRP). Rat DFATs and aPRP were isolated using ceiling culture and centrifugation, respectively. The cell proliferation rate was measured, and the cells were cultured in an osteoblast differentiation medium under varying concentrations of aPRP for 21 days and stained with Alizarin red. Gene expression was evaluated using real time polymerase chain reaction. Critical defects were implanted with DFAT seeded gelatin sponges under aPRP, and four weeks later, the bone regeneration ability was evaluated using micro-computed tomography and hematoxylin-eosin staining. The cell proliferation rate was significantly increased by the addition of aPRP. Alizarin red staining was positive 21 days after the start of induction, with significantly higher Runt-related transcription factor 2 (Runx2) and osteocalcin (OCN) expression levels than those in the controls. A 9 mm critical defect was largely closed (60.6%) after four weeks of gelatin sponge implantation with DFAT and aPRP. Therefore, materials combining DFAT cells and aPRP may be an effective approach for bone regeneration. Further research is needed to explore the long-term effects of these materials.

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Effect of Gelatin on Osteogenic Cell Sheet Formation Using Canine Adipose-Derived Mesenchymal Stem Cells

Cell Transplantation ◽

10.3727/096368916x693338 ◽

2017 ◽

Vol 26 (1) ◽

pp. 115-123 ◽

Cited By ~ 15

Author(s):

Ah Young Kim ◽

Yongsun Kim ◽

Seung Hoon Lee ◽

Yongseok Yoon ◽

Wan-Hee Kim ◽

...

Keyword(s):

Stem Cells ◽

Cell Proliferation ◽

Mesenchymal Stem Cells ◽

Bone Regeneration ◽

Cell Sheet ◽

Proliferation Rate ◽

Osteogenic Cell ◽

Cell Sheets ◽

Gene Markers ◽

Cell Proliferation Rate

Osteogenically differentiated cell sheet techniques using mesenchymal stem cells (MSCs) are available to stimulate bone regeneration. The advantage of the cell sheet technique is delivering live cells effectively into the focal region. We developed a novel osteogenic cell sheet technique by adding gelatin to osteogenic cell medium. Gelatin-induced osteogenic cell sheets (GCSs) were compared to conventional osteogenic cell sheets (OCSs). Undifferentiated MSCs (UCs) were used as a control. The morphology of these cell sheets was evaluated microscopically and histologically. The time-dependent cell proliferation rate was estimated by DNA quantification. The expression of osteogenic gene markers and the number of calcium depositions were assessed by quantitative real-time polymerase chain reaction and Alizarin red S (ARS) staining, respectively. GCSs were thicker and stronger than OCSs. GCSs showed a significantly higher cell proliferation rate compared to OCSs ( p < 0.05). GCSs exhibited significantly higher upregulation of BMP-7 mRNA compared to OCSs ( p < 0.05). Both GCSs and OCSs showed negative ARS reactivity on day 10, but only GCSs showed positive ARS reactivity on day 21. With this technique, we observed active cell proliferation with abundant ECM and upregulation of osteogenic bone markers, and our results suggest that GCSs could be promising for therapeutic applications in bone regeneration.

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Evidence for a Negative Correlation between Human Reactive Enamine-Imine Intermediate Deaminase A (RIDA) Activity and Cell Proliferation Rate: Role of Lysine Succinylation of RIDA

International Journal of Molecular Sciences ◽

10.3390/ijms22083804 ◽

2021 ◽

Vol 22 (8) ◽

pp. 3804

Author(s):

Luisa Siculella ◽

Laura Giannotti ◽

Benedetta Di Chiara Stanca ◽

Matteo Calcagnile ◽

Alessio Rochira ◽

...

Keyword(s):

Cell Proliferation ◽

Cell Lines ◽

Negative Correlation ◽

Cancer Biology ◽

Human Tumor ◽

In Silico Analysis ◽

Proliferation Rate ◽

Reactive Intermediate ◽

Cell Proliferation Rate

Reactive intermediate deaminase (Rid) proteins are enzymes conserved in all domains of life. UK114, a mammalian member of RidA subfamily, has been firstly identified as a component of liver perchloric acid-soluble proteins (L-PSP). Although still poorly defined, several functions have been attributed to the mammalian protein UK114/RIDA, including the reactive intermediate deamination activity. The expression of UK114/RIDA has been observed in some tumors, arousing interest in this protein as an evaluable tumor marker. However, other studies reported a negative correlation between UK114/RIDA expression, tumor differentiation degree and cell proliferation. This work addressed the question of UK114/RIDA expression in human non-tumor HEK293 cell lines and in some human tumor cell lines. Here we reported that human RIDA (hRIDA) was expressed in all the analyzed cell line and subjected to lysine (K-)succinylation. In HEK293, hRIDA K-succinylation was negatively correlated to the cell proliferation rate and was under the control of SIRT5. Moreover, K-succinylation clearly altered hRIDA quantification by immunoblotting, explaining, at least in part, some discrepancies about RIDA expression reported in previous studies. We found that hRIDA was able to deaminate reactive enamine-imine intermediates and that K-succinylation drastically reduced deaminase activity. As predicted by in silico analysis, the observed reduction of deaminase activity has been related to the drastic alterations of hRIDA structure inferred by K-succinylation. The role of hRIDA and the importance of its K-succinylation in cell metabolism, especially in cancer biology, have been discussed.

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Cell proliferation rate and nuclear morphometry in Roberts syndrome

Clinical Genetics ◽

10.1111/j.1399-0004.1998.tb03772.x ◽

2008 ◽

Vol 54 (6) ◽

pp. 512-516 ◽

Cited By ~ 6

Author(s):

Petros M Pavlopoulos ◽

Anastasia E Konstantinidou ◽

Emmanuel Agapitos ◽

Panagiotis Davaris

Keyword(s):

Cell Proliferation ◽

Proliferation Rate ◽

Roberts Syndrome ◽

Nuclear Morphometry ◽

Cell Proliferation Rate

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Analysis of cell proliferation rate in Oral Leukoplakia and Oral Squamous Cell Carcinoma

Journal of Clinical and Experimental Dentistry ◽

10.4317/jced.2.e173 ◽

2010 ◽

pp. e173-177 ◽

Cited By ~ 2

Author(s):

S. Mehkri ◽

AR. Iyengar ◽

KS. Nagesh ◽

MB. Bharati

Keyword(s):

Cell Proliferation ◽

Squamous Cell Carcinoma ◽

Oral Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Squamous Cell ◽

Proliferation Rate ◽

Oral Leukoplakia ◽

Cell Proliferation Rate

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Osteogenic Effects of Triterpene Saponin Soyasapogenol B on Differentiation, Mineralization, Autophagy, and Necroptosis

10.21203/rs.3.rs-1182818/v1 ◽

2021 ◽

Author(s):

Kyung-Ran Park ◽

Joon Yeop Lee ◽

Soo Hyun Kim ◽

Il Keun Kwon ◽

Hyung-Mun Yun

Keyword(s):

Cell Proliferation ◽

Osteoblast Differentiation ◽

Biological Effects ◽

Bone Diseases ◽

Adhesion Assay ◽

Pharmacological Properties ◽

Alizarin Red ◽

Triterpenoid Saponins ◽

Triterpene Saponin ◽

Osteogenic Activity

Abstract Background: Triterpenoid saponins are a diverse group of natural compounds in plants. A triterpene saponin, Soyasapogenol B (SoyB), from Arachis hypogaea (peanut) has various pharmacological properties. This study aimed to elucidate pharmacological properties and mechanisms of SoyB on bone-forming cells. Methods: Cell viability adhesion, and migration were analyzed using MTT assay, cell adhesion assay, and Boyden chamber assay. Osteogenic activity and osteogenicity were analyzed using alkaline phosphatase (ALP) staining and activity, and Alizarin Red S (ARS) staining. Cell signaling, protein expression, and autophagy were analyzed using Western blot analysis, immunofluorescence assay, and DAPGreen autophagy detection assay. Results and Conclusion: In the present study, SoyB (> 99.99% purity), triterpene saponin, was isolated from the fruit of A. hypogaea. At concentrations ranging from 1 to 20 mM, SoyB showed no cell proliferation effects, whereas 30 - 100 mM SoyB increased cell proliferation in MC3T3-E1 cells. Next, osteoblast differentiation was analyzed and found that SoyB enhanced ALP staining and activity and bone mineralization as evidence for early and late osteoblast differentiation. SoyB also induced RUNX2 expression in nucleus with the increased phosphorylation of Smad1/5/8 and JNK2 during osteoblast differentiation. In addition, SoyB-mediated osteoblast differentiation was not associated with autophagy and necroptosis. Furthermore, SoyB increased cell migration and adhesion with the upregulation of MMP13 levels during osteoblast differentiation. The findings of this study provide new evidence that SoyB possesses biological effects on osteogenic activity and osteogenicity in bone-forming cells, and suggest a potentially beneficial role for peanuts foods and drugs containing SoyB in the treatment and prevention of bone diseases.

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Parathyroid gland volume increases with postmaturational aging in the rat

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00261.2001 ◽

2002 ◽

Vol 282 (3) ◽

pp. E557-E563 ◽

Cited By ~ 7

Author(s):

Bernard Halloran ◽

Per Udén ◽

Quan-Yang Duh ◽

Shoichi Kikuchi ◽

Tracy Wieder ◽

...

Keyword(s):

Cell Proliferation ◽

Renal Insufficiency ◽

Parathyroid Gland ◽

Inorganic Phosphorus ◽

Proliferation Rate ◽

Parathyroid Cell ◽

Fischer 344 ◽

Gland Volume ◽

Cell Proliferation Rate ◽

Age Related

To examine the pathophysiology of the age-related rise in the plasma concentration of parathyroid hormone (PTH), we studied the relationships among plasma immunoreactive PTH (iPTH), parathyroid gland volume, parathyroid cell proliferation rate, renal function, and blood Ca2+ in male Fischer 344 rats aged 6–28 mo. Plasma iPTH increased 2.5-fold between 6 and 28 mo and correlated with parathyroid gland volume ( r = 0.87). Gland volume began to increase as early as 6–12 mo of age and by 28 mo was threefold greater than at 6 mo. Gland expansion was a consequence of hyperplasia stimulated in part by an increase in cell proliferative activity late in life. Blood Ca2+ and plasma inorganic phosphorus did not change significantly with age. Glomerular filtration rate decreased with age but only after the age of 24 mo. Unlike what has been observed in the human, these data suggest that the age-related increase in plasma iPTH in the rat is linked to parathyroid gland hyperplasia and that early gland growth does not appear to be associated with hypocalcemia or renal insufficiency, but rather to developmentally related metabolic changes. Later in life (>24 mo), the increase in parathyroid cell proliferation rate, further hyperplastic expansion of the gland, and increase in iPTH secretion appear to be associated with renal insufficiency.

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The Novel Long Noncoding RNA TUSC7 Inhibits Proliferation by Sponging MiR-211 in Colorectal Cancer

Cellular Physiology and Biochemistry ◽

10.1159/000457938 ◽

2017 ◽

Vol 41 (2) ◽

pp. 635-644 ◽

Cited By ~ 57

Author(s):

Jian Xu ◽

Rui Zhang ◽

Jian Zhao

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Tumor Suppressor ◽

Long Noncoding Rna ◽

Noncoding Rna ◽

Proliferation Rate ◽

Expression Level ◽

The Novel ◽

Cell Proliferation Rate ◽

Potential Tumor Suppressor

Background/Aims: The novel long noncoding RNA (lncRNA) tumor suppressor candidate 7 (TUSC7) has been reported as a potential tumor suppressor, while the functional role of TUSC7 is still unknown in colorectal cancer (CRC). Here, we characterized TUSC7 expression profile in CRC patients and investigated its biological function and potential molecular mechanism. Methods: RNA isolation, qRT-PCR, cell counter kit-8 assay, cell cycle assay, EdU assay, and western blot were performed. Statistical analyses were performed using SPSS 18.0 software and p value < 0.05 was considered as statistically significant. Results: In a cohort of CRC patients, we found TUSC7 was significantly downregulated in CRC tissues compared with adjacent non-tumor tissues (P < 0.01). Patients with high expression of TUSC7 had better survival than those with low expression of TUSC7 (HR = 0.342, 95% CI: 0.120-0.972, P = 0.044). Cell count kit 8 and EdU assays showed that ectopic expression of TUSC7 in HCT116 and SW480 cells significantly inhibited cell proliferation rate. After silence of TUSC7 with small interfering RNA, cell proliferation rate increased. Flow cytometry analyses revealed cycles were arrested at G1 phase after TUSC7 overexpression. We found there were 2 binding sites of miR-211-3p within the sequence of TUSC7 and TUSC7 expression level was negatively correlated with miR-211-3p. TUSC7 overexpression increased the expression level of CDK6, which is a downstream target of miR-211-3p, in both RNA and protein level. Furthermore, luciferase reporter assay indicated that TUSC7 could sponge miR-211-3p. Conclusion: To summary, we demonstrated that TUSC7 is a potential tumor suppressor in CRC, and TUSC7 could inhibit CRC cell proliferation by completely sponging miR-211-3p.

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