Establishment of human immortalized mesenchymal stem cells lines for the monitoring and analysis of osteogenic differentiation in living cells

Narai, Takashi; Watase, Ryohei; Nakayama, Yuji; Kodani, Isamu; Inoue, Toshiaki; Kokura, Kenji

doi:10.1016/j.heliyon.2020.e05398

Cited by 5 publications

(5 citation statements)

References 25 publications

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“…B. CRISPR-Cas9 eingesetzt 78 . Dadurch soll ein effizienter klinischer Einsatz von MSCs mit stabilen Behandlungsergebnissen und einer kürzeren Behandlungsdauer bei Knochenregenerationsbehandlungen erreicht werden 78 .…”

Section: Ausblickunclassified

“…Verschiedene Tissue-Engineering-Ansätze zielen auf die Kombination von präkonditionierten MSCs mit Scaffolds zur Geweberegeneration ab. Wegen diverser praktischer Herausforderungen, wie der Isolation, Expansion und Applikation der MSCs, und aufgrund ihrer begrenzten Effizienz an der Transplantationsstelle werden sie noch nicht in großem Umfang für die Knochenregeneration eingesetzt 78 .…”

Section: Ausblickunclassified

“…Um die osteogenen Differenzierungsprozesse reproduzierbarer und kontrollierbarer zu machen, werden neuartige Techniken zur Genom-Editierung wie z. B. CRISPR-Cas9 eingesetzt 78 . Dadurch soll ein effizienter klinischer Einsatz von MSCs mit stabilen Behandlungsergebnissen und einer kürzeren Behandlungsdauer bei Knochenregenerationsbehandlungen erreicht werden 78 .…”

Section: Ausblickunclassified

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Knochenaufbau und Knochenersatzmaterialien

Lang¹,

Klute²,

Rupp³

et al. 2022

Orthopädie und Unfallchirurgie up2date

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Knochendefekte können primär aus traumatologischen Ursachen sowie sekundär aus Knocheninfektionen und Tumorerkrankungen resultieren. Die chirurgische Wiederherstellung der Bewegungsorgane bei Vorliegen eines behandlungsbedürftigen Knochendefekts stellt trotz der Entwicklung diverser Verfahren zum Knochenersatz und Knochenaufbau auch heute noch eine Herausforderung dar. Das autologe Knochentransplantat gilt als Goldstandard in der Behandlung. Wachsende Bedeutung wird den synthetischen Knochenersatzmaterialien beigemessen.

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Section: Ausblickunclassified

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Knochenaufbau und Knochenersatzmaterialien

Lang¹,

Klute²,

Rupp³

et al. 2022

Orthopädie und Unfallchirurgie up2date

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“…Numerous authors have concentrated their attention on the application of PRF in the tissue engineering of cartilage and tendons, which has resulted in an increase in in vitro, preclinical, and clinical research [ 14 , 19 , 20 ]. Chondrogenesis depends on several cartilage-specific markers, including collagen, aggrecan (ACAN), SRY-box transcription factor 9 (SOX9), and osteogenic-related markers, such as alkaline phosphatase (ALPL), bone gamma-carboxyglutamate protein (BGLAP), and RUNX family transcription factor 2 (RUNX2), which participate in tissue regeneration [ 21 , 22 , 23 , 24 , 25 ].…”

Section: Introductionmentioning

confidence: 99%

Osteochondral Regeneration Ability of Uncultured Bone Marrow Mononuclear Cells and Platelet-Rich Fibrin Scaffold

Nguyen-Thanh,

Nguyen-Tran,

Cruciani

et al. 2023

Bioengineering

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Objectives: Platelet-rich fibrin (PRF) and bone marrow mononuclear cells are potential scaffolds and cell sources for osteochondral regeneration. The main aim of this paper is to examine the effects of PRF scaffolds and autologous uncultured bone marrow mononuclear cells on osteochondral regeneration in rabbit knees. Materials and Methods: Three different types of PRF scaffolds were generated from peripheral blood (Ch-PRF and L-PRF) and bone marrow combined with uncultured bone marrow mononuclear cells (BMM-PRF). The histological characteristics of these scaffolds were assessed via hematoxylin–eosin staining, PicroSirius red staining, and immunohistochemical staining. Osteochondral defects with a diameter of 3 mm and depth of 3 mm were created on the trochlear groove of the rabbit’s femur. Different PRF scaffolds were then applied to treat the defects. A group of rabbits with induced osteochondral defects that were not treated with any scaffold was used as a control. Osteochondral tissue regeneration was assessed after 2, 4, and 6 weeks by macroscopy (using the Internal Cartilage Repair Society score, X-ray) and microscopy (hematoxylin—eosin stain, safranin O stain, toluidine stain, and Wakitani histological scale, immunohistochemistry), in addition to gene expression analysis of osteochondral markers. Results: Ch-PRF had a heterogeneous fibrin network structure and cellular population; L-PRF and BMM-PRF had a homogeneous structure with a uniform distribution of the fibrin network. Ch-PRF and L-PRF contained a population of CD45-positive leukocytes embedded in the fibrin network, while mononuclear cells in the BMM-PRF scaffold were positive for the pluripotent stem cell-specific antibody Oct-4. In comparison to the untreated group, the rabbits that were given the autologous graft displayed significantly improved healing of the articular cartilage tissue and of the subchondral bone. Regeneration was gradually observed after 2, 4, and 6 weeks of PRF scaffold treatment, which was particularly evident in the BMM-PRF group. Conclusions: The combination of biomaterials with autologous platelet-rich fibrin and uncultured bone marrow mononuclear cells promoted osteochondral regeneration in a rabbit model more than platelet-rich fibrin material alone. Our results indicate that autologous platelet-rich fibrin scaffolds combined with uncultured bone marrow mononuclear cells applied in healing osteochondral lesions may represent a suitable treatment in addition to stem cell and biomaterial therapy.

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“…Other systems applied the CRISPR/Cas9 system for the live imaging of proteins, guided by the sgRNA to locate specific regions on the genome ( Ma et al, 2018 ) as done by Narai et al in which they used CRISPR technology to localize osteogenic differentiation in MSCs through the monitoring of bone gamma-carboxyglutamate protein (BGLAP) expression in vivo via an enhanced green fluorescent protein (EGFP) reporter ( Narai et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

An Overview of RNA-Based Scaffolds for Osteogenesis

Damiati

El-Messeiry

2021

Front. Mol. Biosci.

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Tissue engineering provides new hope for the combination of cells, scaffolds, and bifactors for bone osteogenesis. This is achieved by mimicking the bone’s natural behavior in recruiting the cell’s molecular machinery for our use. Many researchers have focused on developing an ideal scaffold with specific features, such as good cellular adhesion, cell proliferation, differentiation, host integration, and load bearing. Various types of coating materials (organic and non-organic) have been used to enhance bone osteogenesis. In the last few years, RNA-mediated gene therapy has captured attention as a new tool for bone regeneration. In this review, we discuss the use of RNA molecules in coating and delivery, including messenger RNA (mRNA), RNA interference (RNAi), and long non-coding RNA (lncRNA) on different types of scaffolds (such as polymers, ceramics, and metals) in osteogenesis research. In addition, the effect of using gene-editing tools—particularly CRISPR systems—to guide RNA scaffolds in bone regeneration is also discussed. Given existing knowledge about various RNAs coating/expression may help to understand the process of bone formation on the scaffolds during osseointegration.

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Establishment of human immortalized mesenchymal stem cells lines for the monitoring and analysis of osteogenic differentiation in living cells

Cited by 5 publications

References 25 publications

Knochenaufbau und Knochenersatzmaterialien

Knochenaufbau und Knochenersatzmaterialien

Osteochondral Regeneration Ability of Uncultured Bone Marrow Mononuclear Cells and Platelet-Rich Fibrin Scaffold

An Overview of RNA-Based Scaffolds for Osteogenesis

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