Role of FBXW2 in explant cultures of bovine periosteum-derived cells

Abstract Objective Bone regeneration is a potential technique for treating osteoporosis. A previous study reported that F-box and WD-40 domain-containing protein 2 (FBXW2) localized with osteocalcin in bovine periosteum after 5 weeks of explant culture. However, the osteoblastic functions of FBXW2 remain unclear. In this study, double-fluorescent immunostaining was used to investigate the potential role of FBXW2 and its relationship with osteocalcin. Results At day 0, FBXW2 was expressed in the cambium layer between the bone and periosteum, while osteocalcin was expressed in bone. After explant culture, changes in the periosteum were observed from weeks 1 to 7. At week 1, partial FBXW2 expression was seen with a small amount of osteocalcin. At week 2, a layer of FBXW2 was observed. From weeks 3 to 7, tube-like structures of FBXW and osteocalcin were observed, and periosteum-derived cells were released from the periosteum in areas where no FBXW2 was observed. Bovine periosteum-derived cells can form a three-dimensional cell pellet, because multilayered cell sheets are formed inside of the periosteum in vitro. It is shown that in results FBXW2 is produced in periosteal explants near sites where initial osteogenic activity is observed, suggesting that it may be involved in periosteal osteogenesis.

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In Vitro Comparison of 2D-Cell Culture and 3D-Cell Sheets of Scleraxis-Programmed Bone Marrow Derived Mesenchymal Stem Cells to Primary Tendon Stem/Progenitor Cells for Tendon Repair

International Journal of Molecular Sciences ◽

10.3390/ijms19082272 ◽

2018 ◽

Vol 19 (8) ◽

pp. 2272 ◽

Cited By ~ 10

Author(s):

Chi-Fen Hsieh ◽

Zexing Yan ◽

Ricarda Schumann ◽

Stefan Milz ◽

Christian Pfeifer ◽

...

Keyword(s):

Bone Marrow ◽

Cell Sheet ◽

Tendon Repair ◽

Three Dimensional ◽

Cell Types ◽

Tendon Tissue ◽

Cell Sheets ◽

Tendon Tissue Engineering ◽

Dimensional Cell

The poor and slow healing capacity of tendons requires novel strategies to speed up the tendon repair process. Hence, new and promising developments in tendon tissue engineering have become increasingly relevant. Previously, we have established a tendon progenitor cell line via ectopic expression of the tendon-related basic helix-loop-helix (bHLH) transcription factor Scleraxis (Scx) in human bone marrow mesenchymal stem cells (hMSC-Scx). The aim of this study was to directly compare the characteristics of hMSC-Scx cells to that of primary human tendon stem/progenitors cells (hTSPCs) via assessment of self-renewal and multipotency, gene marker expression profiling, in vitro wound healing assay and three-dimensional cell sheet formation. As expected, hTSPCs were more naive than hMSC-Scx cells because of higher clonogenicity, trilineage differentiation potential, and expression of stem cell markers, as well as higher mRNA levels of several gene factors associated with early tendon development. Interestingly, with regards to wound healing, both cell types demonstrate a comparable speed of scratch closure, as well as migratory velocity and distance in various migration experiments. In the three-dimensional cell sheet model, hMSC-Scx cells and hTSPCs form compact tendinous sheets as histological staining, and transmission electron microscopy shows spindle-shaped cells and collagen type I fibrils with similar average diameter size and distribution. Taken together, hTSPCs exceed hMSC-Scx cells in several characteristics, namely clonogenicity, multipotentiality, gene expression profile and rates of tendon-like sheet formation, whilst in three-dimensional cell sheets, both cell types have comparable in vitro healing potential and collagenous composition of their three-dimensional cell sheets, making both cell types a suitable cell source for tendon tissue engineering and healing.

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Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010005161x ◽

1974 ◽

Vol 32 ◽

pp. 320-321

Author(s):

J. P. Revel

Keyword(s):

Developmental Stages ◽

Three Dimensional ◽

Cell Movement ◽

Cellular Interactions ◽

Serial Sections ◽

Cell Sheets ◽

Freeze Etching ◽

Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

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Specificity of the HIV-1 Protease on Substrates Representing the Cleavage Site in the Proximal Zinc-Finger of HIV-1 Nucleocapsid Protein

Viruses ◽

10.3390/v13061092 ◽

2021 ◽

Vol 13 (6) ◽

pp. 1092

Author(s):

János András Mótyán ◽

Márió Miczi ◽

Stephen Oroszlan ◽

József Tőzsér

Keyword(s):

Amino Acid ◽

Zinc Finger ◽

Cleavage Site ◽

Potential Role ◽

Binding Mode ◽

Interaction Energies ◽

Vitro Assay ◽

Hiv 1

To explore the sequence context-dependent nature of the human immunodeficiency virus type 1 (HIV-1) protease’s specificity and to provide a rationale for viral mutagenesis to study the potential role of the nucleocapsid (NC) processing in HIV-1 replication, synthetic oligopeptide substrates representing the wild-type and modified versions of the proximal cleavage site of HIV-1 NC were assayed as substrates of the HIV-1 protease (PR). The S1′ substrate binding site of HIV-1 PR was studied by an in vitro assay using KIVKCF↓NCGK decapeptides having amino acid substitutions of N17 residue of the cleavage site of the first zinc-finger domain, and in silico calculations were also performed to investigate amino acid preferences of S1′ site. Second site substitutions have also been designed to produce “revertant” substrates and convert a non-hydrolysable sequence (having glycine in place of N17) to a substrate. The specificity constants obtained for peptides containing non-charged P1′ substitutions correlated well with the residue volume, while the correlation with the calculated interaction energies showed the importance of hydrophobicity: interaction energies with polar residues were related to substantially lower specificity constants. Cleavable “revertants” showed one residue shift of cleavage position due to an alternative productive binding mode, and surprisingly, a double cleavage of a substrate was also observed. The results revealed the importance of alternative binding possibilities of substrates into the HIV-1 PR. The introduction of the “revertant” mutations into infectious virus clones may provide further insights into the potential role of NC processing in the early phase of the viral life-cycle.

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The Role of Pre-Clinical 3-Dimensional Models of Osteosarcoma

International Journal of Molecular Sciences ◽

10.3390/ijms21155499 ◽

2020 ◽

Vol 21 (15) ◽

pp. 5499

Author(s):

Hannah L. Smith ◽

Stephen A. Beers ◽

Juliet C. Gray ◽

Janos M. Kanczler

Keyword(s):

Three Dimensional ◽

Chorioallantoic Membrane ◽

3D Models ◽

In Ovo ◽

Future Directions ◽

3 Dimensional ◽

In Vivo Animal Models

Treatment for osteosarcoma (OS) has been largely unchanged for several decades, with typical therapies being a mixture of chemotherapy and surgery. Although therapeutic targets and products against cancer are being continually developed, only a limited number have proved therapeutically active in OS. Thus, the understanding of the OS microenvironment and its interactions are becoming more important in developing new therapies. Three-dimensional (3D) models are important tools in increasing our understanding of complex mechanisms and interactions, such as in OS. In this review, in vivo animal models, in vitro 3D models and in ovo chorioallantoic membrane (CAM) models, are evaluated and discussed as to their contribution in understanding the progressive nature of OS, and cancer research. We aim to provide insight and prospective future directions into the potential translation of 3D models in OS.

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Advanced in vitro management of three-dimensional cell cultures and explanted tissue

Cytotherapy ◽

10.1016/s1465324921005156 ◽

2021 ◽

Vol 23 (5) ◽

pp. S145

Author(s):

S. Kress ◽

D. Egger ◽

C. Kasper

Keyword(s):

Cell Cultures ◽

Three Dimensional ◽

Dimensional Cell

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The potential role of three-dimensional surface imaging as a tool to evaluate aesthetic outcome after Breast Conserving Therapy (BCT)

Breast Cancer Research and Treatment ◽

10.1007/s10549-017-4256-y ◽

2017 ◽

Vol 164 (2) ◽

pp. 385-393 ◽

Cited By ~ 6

Author(s):

Rachel L. O’Connell ◽

Rosa Di Micco ◽

Komel Khabra ◽

Lisa Wolf ◽

Nandita deSouza ◽

...

Keyword(s):

Potential Role ◽

Three Dimensional ◽

Breast Conserving Therapy ◽

Surface Imaging ◽

Aesthetic Outcome ◽

Dimensional Surface

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A three-dimensional cell biology model of human hepatocellular carcinoma in vitro

Tumor Biology ◽

10.1007/s13277-010-0140-7 ◽

2010 ◽

Vol 32 (3) ◽

pp. 469-479 ◽

Cited By ~ 15

Author(s):

Jianhua Tang ◽

Jiefeng Cui ◽

Rongxin Chen ◽

Kun Guo ◽

Xiaonan Kang ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Biology ◽

Three Dimensional ◽

Human Hepatocellular Carcinoma ◽

Dimensional Cell

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Anterior mesendoderm induces mouse Engrailed genes in explant cultures

Development ◽

10.1242/dev.118.1.139 ◽

1993 ◽

Vol 118 (1) ◽

pp. 139-149 ◽

Cited By ~ 36

Author(s):

S.L. Ang ◽

J. Rossant

Keyword(s):

Direct Evidence ◽

Neural Tissue ◽

Explant Culture ◽

Neural Plate ◽

Explant Cultures ◽

Neural Marker ◽

Engrailed Genes ◽

Anterior Posterior

We have developed germ layer explant culture assays to study the role of mesoderm in anterior-posterior (A-P) patterning of the mouse neural plate. Using isolated explants of ectodermal tissue alone, we have demonstrated that the expression of Engrailed-1 (En-1) and En-2 genes in ectoderm is independent of mesoderm by the mid- to late streak stage, at least 12 hours before their onset of expression in the neural tube in vivo at the early somite stage. In recombination explants, anterior mesendoderm from headfold stage embryos induces the expression of En-1 and En-2 in pre- to early streak ectoderm and in posterior ectoderm from headfold stage embryos. In contrast, posterior mesendoderm from embryos of the same stage does not induce En genes in pre- to early streak ectoderm but is able to induce expression of a general neural marker, neurofilament 160 × 10(3) M(r). These results provide the first direct evidence for a role of mesendoderm in induction and regionalization of neural tissue in mouse.

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High Expression of GSDMC Is Associated with Poor Survival in Kidney Clear Cell Cancer

BioMed Research International ◽

10.1155/2021/5282894 ◽

2021 ◽

Vol 2021 ◽

pp. 1-10

Author(s):

Yun-Qian Cui ◽

Fei Meng ◽

Wen-Li Zhan ◽

Zhou-Tong Dai ◽

Xinghua Liao

Keyword(s):

Potential Role ◽

Clear Cell ◽

Cell Cancer ◽

Progression Free Survival ◽

Renal Clear Cell Carcinoma ◽

High Expression ◽

Poor Survival ◽

Free Survival

This study is aimed at exploring the potential role of GSDMC in kidney renal clear cell carcinoma (KIRC). We analyzed the expression of GSDMC in 33 types of cancers in TCGA database. The results showed that the expression of GSDMC was upregulated in most cancers. We found a significant association between high expression of GSDMC and shortened patient overall survival, progression-free survival, and disease-specific survival. In vitro experiments have shown that the expression of GSDMC was significantly elevated in KIRC cell lines. Moreover, decreased expression of GSDMC was significantly associated with decreased cell proliferation. In summary, we believe that this study provides valuable data supporting future clinical treatment.

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