In vitro human periodontal ligament-like tissue formation with porous poly-l-lactide matrix

Liao, Wen; Okada, Masahiro; Sakamoto, Fumito; Okita, Naoya; Inami, Kaoru; Nishiura, Aki; Hashimoto, Yoshiya; Matsumoto, Naoyuki

doi:10.1016/j.msec.2013.04.008

Cited by 12 publications

(11 citation statements)

References 32 publications

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“…Runx-2 is a key gene marker of HPdLF metabolism within bone, and the 356 expression of this gene often serves as an early marker for osteogenic activity. OCN is an 357 osteogenic marker gene that serves as a mid-range marker of bone metabolism (Oortgiesen et al (Liao et al 2013). In this study, a stable increase occurred in the COL-3 368 gene in both PN and PF groups during the 7-day stretching period.…”

supporting

confidence: 51%

Peer Review #1 of "Accelerated construction of an in vitro model of human periodontal ligament tissue: vacuum plasma combined with fibronectin coating and a polydimethylsiloxane matrix (v0.1)"

2019

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Tying shape memory wires to crowded teeth causes the wires to deform according to the dental arch. This deformation results in a resilient force that is delivered to the tooth. The appropriate amount of force can activate the osteogenetic and osteoclastic ability of the periodontal ligament (PDL) and the tooth can be moved. This is the biological basis of orthodontic treatment. To achieve further insight into the mechanisms underlying orthodontic treatment, we examined whether accelerated construction of an in vitro human PDL fibroblast (HPdLF) stretching model can be achieved by combining fibronectin coating and vacuum plasma treatment with polydimethylsiloxane (PDMS) cell-culture chambers. Each chamber was randomly assigned to a no-surface modification (NN), fibronectin coating (FN), vacuum plasma treatment (PN), or vacuum plasma treatment followed by a fibronectin coating (PF) treatment protocol. The physical and chemical features and ability to promote cellular proliferation of the PDMS chamber surfaces were evaluated. Cellular adhesion of four materials were evaluated and two best-proliferated groups were considered as better model-constructing surfaces and used in subsequent experiments and used in subsequent experiments. HPdLFs were cultured on these two kinds of chambers without stretching for 3 days, then with stretching for 7 days. Timecourse gene expression cellular morphology were evaluated. Chambers in the PN group had high wettability and surface component changes. The FN and PF chambers had high cellular proliferation ability. They were selected into subsequent experiments. After 3 days of culturing HPdLFs on the PF and PN chambers, the cells in the PF chambers had PeerJ reviewing PDF | Manuscript to be reviewed significantly higher levels of runt-related transcription factor 2 (Runx-2) and osteocalcin (OCN) gene expression compared with the cells in the PN chambers. After cyclic stretch application to the cells in the PN and PF chambers, expression of the type-3 collagen (COL-3) gene in PF group continued to increase for 7 days and was significantly higher than that in the PN group from day 5 onwards. The HPdLFs in the PF group showed parallel alignment from days 3 to 7 after imposition of cyclic stretch, while those in the PN group aligned in parallel from day 5 on. Our results suggested that applying a fibronectin coating to a PDMS chamber after plasma treatment can accelerate establishment of an in vitro PDL stretching model.PeerJ reviewing PDF |

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confidence: 51%

Peer Review #1 of "Accelerated construction of an in vitro model of human periodontal ligament tissue: vacuum plasma combined with fibronectin coating and a polydimethylsiloxane matrix (v0.1)"

2019

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“…A major limitation of culture in a matrix is the limited cell survival to 3 or 4 days. Only the poly-l-lactide matrix allows a longer follow-up, although limited to 2 weeks (32). This is why we did not carry out a follow-up on a longer term.…”

Section: Discussionmentioning

confidence: 99%

Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics

Brézulier

Pellen‐Mussi²,

Tricot‐Doleux³

et al. 2020

European Journal of Orthodontics

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Summary Objectives Mechanobiology phenomena constitute a major element of the cellular and tissue response during orthodontic treatment and the implantation of a biomaterial. Better understanding these phenomena will improve the effectiveness of our treatments. The objective of this work is to validate a model of three-dimensional (3D) culture of osteoblasts to study mechanobiology. Materials and methods The hFOB 1.19 cell line was cultured either traditionally on a flat surface or in aggregates called spheroids. They were embedded in 0.8% low-melting agarose type VII and placed in a polyethylene terephthalate transwell insert. Compressive forces of 1 and 4 g/cm2 were applied with an adjustable weight. Proliferation was evaluated by measuring diameters, monitoring glucose levels, and conducting Hoechst/propidium iodide staining. Enzyme-linked immunosorbent assays focusing on the pro-inflammatory mediators interleukin (IL)-6 and IL-8 and bone remodelling factor osteoprotegerin were performed to evaluate soluble factor synthesis. quantitative reverse transcription-polymerase chain reaction was performed to evaluate bone marker transcription. Results The 3D model shows good cell viability and permits IL dosing. Additionally, three gene expression profiles are analysable. Limitations The model allows analysis of conventional markers; larger exploration is needed for better understanding osteoblast mechanobiology. However, it only allows an analysis over 3 days. Conclusion The results obtained by applying constant compressive forces to 3D osteoblastic cultures validate this model system for exploring biomolecule release and analysing gene transcription. In particular, it highlights a disturbance in the expression of markers of osteogenesis.

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“…The in vitro HPdLLT was prepared according a protocol published previously (Liao et al 2013). Briefly, PLLA pellets (average MW = 240,000; BMG, Kyoto, Japan) were dissolved in dichloromethane (DCM; 5 % wt/v; Nacalai Tesque, Kyoto, Japan) and cast with 75-150 lm sized sodium chloride granules (NaCl; Nacalai Tesque) as a porogen.…”

Section: Preparation Of In Vitro Three-dimensional Human Periodontal mentioning

confidence: 99%

“…Recently, we developed an in vitro human periodontal ligament-like tissue (HPdLLT) model by 3D culture of human periodontal ligament fibroblasts in a porous poly-L-lactide matrix. This matrix was hydrophilically modified using ammonia solution, enabling threefold greater permeability of culture media (Liao et al 2013). Higher culture media permeability directly influences cell living conditions inside the matrix, allowing greater attachment of cells to the 3D matrix in the early stages of cell growth and more accessibility to nutrients and oxygen inside the matrix.…”

Section: Introductionmentioning

confidence: 99%

Cell survival and gene expression under compressive stress in a three-dimensional in vitro human periodontal ligament-like tissue model

et al. 2014

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This study investigated cell survival and gene expression under various compressive stress conditions mimicking orthodontic force by using a newly developed in vitro model of human periodontal ligament-like tissue (HPdLLT). The HPdLLT was developed by three-dimensional culturing of human periodontal ligament fibroblasts in a porous poly-L-lactide matrix with threefold increased culture media permeability due to hydrophilic modification. In vitro HPdLLTs in experimental groups were subjected to 5, 15, 25 and 35 g/cm(2) compressive stress for 1, 3, 7 or 14 days; controls were cultured over the same periods without compressive stress. Cell morphology and cell apoptosis in the experimental and control groups were investigated using scanning electron microscopy and caspase-3/7 detection. Real-time polymerase chain reaction was performed for seven osteogenic and osteoclastic genes. Similar extracellular matrix and spindle-shaped cells were observed inside or on the surface of in vitro HPdLLTs, with no relation to compressive stress duration or intensity. Similar caspase-3/7 activity indicating comparable apoptosis levels was observed in all samples. Receptor activator of nuclear factor kappa-B ligand and bone morphogenetic protein 2 genes showed characteristic "double-peak" expression at 15 and 35 g/cm(2) on day 14, and alkaline phosphatase and periodontal ligament-associated protein 1 expression peaked at 5 g/cm(2) on day 14; other genes also showed time-dependent and load-dependent expression patterns. The in vitro HPdLLT model system effectively mimicked the reaction and gene expression of the human periodontal ligament in response to orthodontic force. This work provides new information on the effects of compressive stress on human periodontal ligament tissue.

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In vitro human periodontal ligament-like tissue formation with porous poly-l-lactide matrix

Cited by 12 publications

References 32 publications

Peer Review #1 of "Accelerated construction of an in vitro model of human periodontal ligament tissue: vacuum plasma combined with fibronectin coating and a polydimethylsiloxane matrix (v0.1)"

Peer Review #1 of "Accelerated construction of an in vitro model of human periodontal ligament tissue: vacuum plasma combined with fibronectin coating and a polydimethylsiloxane matrix (v0.1)"

Development of a 3D human osteoblast cell culture model for studying mechanobiology in orthodontics

Cell survival and gene expression under compressive stress in a three-dimensional in vitro human periodontal ligament-like tissue model

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