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Bone marrow mesenchymal stem cells combine with Treated dentin matrix to build biological root.

ABSTRACT: Treated dentin matrix (TDM) as a kind of scaffolding material has been proved odontogenic induction ability on dental-derived stem cells. Given the limited resources of dental stem cells, it is necessary to seek new seed cell which easily obtained. Jaw bone marrow mesenchymal stem cell (JBMMSC) as non-dental-derived stem cell relates to the development of teeth and jaws which suggest us JBMMSCs could act as a new seed cell for tooth tissue engineering. To assess the odontogenic and osteogenic potential of JBMMSCs, cells were induced by TDM extraction in vitro and combined with TDM in vivo. Results were analyzed by PCR, Western Blotting and histology. PCR and Western Blotting showed odontogenic and osteogenic makers were significantly enhanced in varying degrees after induced by TDM extraction in vitro. In vivo, JBMMSCs expressed both odontogenic and osteogenic-related protein, and the latter showed stronger positive expression. Furthermore, histological examination of the harvested grafts was observed the formation of bone-like tissue. Therefore, osteogenic differentiation ability of JBMMSCs were enhanced significantly after being inducted by TDM which illustrates that non-odontogenic derived stem cells are still promising seed cells in tooth root tissue engineering.

SUBMITTER: Luo S

PROVIDER: S-EPMC5388852 | biostudies-other | 2017 Apr

REPOSITORIES: biostudies-other

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Bone marrow mesenchymal stem cells combine with Treated dentin matrix to build biological root.

Luo Shiwei S Pei Fei F Zhang Wen W Guo Weihua W Li Rui R He Wei W Tian Weidong W

Scientific reports 20170412

Treated dentin matrix (TDM) as a kind of scaffolding material has been proved odontogenic induction ability on dental-derived stem cells. Given the limited resources of dental stem cells, it is necessary to seek new seed cell which easily obtained. Jaw bone marrow mesenchymal stem cell (JBMMSC) as non-dental-derived stem cell relates to the development of teeth and jaws which suggest us JBMMSCs could act as a new seed cell for tooth tissue engineering. To assess the odontogenic and osteogenic po ...[more]

PMID: 28401887

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Project description:UnlabelledMesenchymal stem cells (MSCs) can differentiate into multiple lineages through guidance from the biophysical and biochemical properties of the extracellular matrix. In this work we conduct a combinatorial study of matrix properties that influence adipogenesis and neurogenesis including: adhesion proteins, stiffness, and cell geometry, for mesenchymal stem cells derived from adipose tissue (AT-MSCs) and bone marrow (BM-MSCs). We uncover distinct differences in integrin expression, the magnitude of traction stress, and lineage specification to adipocytes and neuron-like cells between cell sources. In the absence of media supplements, adipogenesis in AT-MSCs is not significantly influenced by matrix properties, while the converse is true in BM-MSCs. Both cell types show changes in the expression of neurogenesis markers as matrix cues are varied. When cultured on laminin conjugated microislands of the same adhesive area, BM-MSCs display elevated adipogenesis markers, while AT-MSCs display elevated neurogenesis markers; integrin analysis suggests neurogenesis in AT-MSCs is guided by adhesion through integrin αvβ3. Overall, the properties of the extracellular matrix guides MSC adhesion and lineage specification to different degrees and outcomes, in spite of their similarities in general characteristics. This work will help guide the selection of MSCs and matrix components for applications where high fidelity of differentiation outcome is desired.Statement of significanceMesenchymal stem cells (MSCs) are an attractive cell type for stem cell therapies; however, in order for these cells to be useful in medicine, we need to understand how they respond to the physical and chemical environments of tissue. Here, we explore how two promising sources of MSCs-those derived from bone marrow and from adipose tissue-respond to the compliance and composition of tissue using model extracellular matrices. Our results demonstrate a source-specific propensity to undergo adipogenesis and neurogenesis, and uncover a role for adhesion, and the degree of traction force exerted on the substrate in guiding these lineage outcomes.

Project description:ObjectivesDentin, the bulk material of the tooth, resemble the bone's chemical composition and is considered a valuable bone substitute. In the current study, we assessed the cytotoxicity and osteogenic potential of demineralized dentin matrix (DDM) in comparison to HA nanoparticles (n-HA) on bone marrow mesenchymal stem cells (BMMSCs) using a hydrogel formulation.Materials and methodsHuman extracted teeth were minced into particles and treated via chemical demineralization using ethylene diamine tetra-acetic acid solution (EDTA) to produce DDM particles. DDM and n-HA particles were added to the sodium alginate then, the combination was dripped into a 5% (w/v) calcium chloride solution to obtain DDM hydrogel (DDMH) or nano-hydroxyapatite hydrogel (NHH). The particles were evaluated by dynamic light scattering (DLS) and the hydrogels were evaluated via scanning electron microscope (SEM). BMMSCs were treated with different hydrogel concentrations (25%, 50%, 75% and neat/100%) and cell viability was evaluated using MTT assay after 72 h of culture. Collagen-I (COL-I) gene expression was studied with real-time quantitative polymerase chain reaction (RT-qPCR) after 3 weeks of culture and alkaline phosphatase (ALP) activity was assessed using enzyme-linked immune sorbent assay (ELISA) over 7th, 10th, 14th and 21st days of culture. BMMSCs seeded in a complete culture medium were used as controls. One-way ANOVA was utilized to measure the significant differences in the tested groups.ResultsDLS measurements revealed that DDM and n-HA particles had negative values of zeta potential. SEM micrographs showed a porous microstructure of the tested hydrogels. The viability results revealed that 100% concentrations of either DDMH or NHH were cytotoxic to BMMSCs after 72 h of culture. However, the cytotoxicity of 25% and 50% concentrations of DDMH were not statistically significant compared to the control group. RT-qPCR showed that COL-I gene expression was significantly upregulated in BMMSCs cultured with 50% DDMH compared to all other treated or control groups (P < 0.01). ELISA analysis revealed that ALP level was significantly increased in the groups treated with 50% DDMH compared to 50% NHH after 21 days in culture (P < 0.001).ConclusionThe injectable hydrogel containing demineralized dentin matrix was successfully formulated. DDMH has a porous structure and has been shown to provide a supporting matrix for the viability and differentiation of BMMSCs. A 50% concentration of DDMH was revealed to be not cytotoxic to BMMSCs and may have a great potential to promote bone formation ability.

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Bone marrow mesenchymal stem cells combine with Treated dentin matrix to build biological root.

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Bone marrow mesenchymal stem cells combine with Treated dentin matrix to build biological root.

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