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Alastair James Sloan - One of the best experts on this subject based on the ideXlab platform.

  • liposomal delivery of demineralized dentin matrix for Dental Tissue Regeneration
    Tissue Engineering Part A, 2018
    Co-Authors: G Melling, John S Colombo, Steven J Avery, Wayne Nishio Ayre, Samuel Lewin Evans, Rachel J Waddington, Alastair James Sloan
    Abstract:

    Current Dental restorations have short longevity, consequently there is a need for novel Tissue engineering strategies that aim to regenerate the dentine-pulp complex. Dentine matrix contains a myriad of bioactive growth factors and extracellular matrix proteins associated with the recruitment, proliferation and differentiation of Dental pulp progenitor cells. Here, we show that demineralised dentine matrix (DDM), from non-carious dentine, can be encapsulated into liposomes for delivery to Dental Tissue to promote Regeneration. Liposomes were formulated to encapsulate 0 - 100 μg/mL DDM, lysed with Triton X and used in VEGF and TGF-β1 ELISAs to quantify release. The encapsulation efficiency was calculated to be 25.9% and 28.8% (VEGF/TGF-β1) for 50 μg/mL DDM liposomes and 39% and 146.7% (VEGF/TGF-β1) for 100 μg/mL DDM liposomes. All liposome formulations had no cytotoxic effects on a Dental pulp stem cell (DPSC) clone, as shown by MTT, Caspase 3/7 assays and cell counts. The ability of the liposomes to stimulate DPSC chemotactic recruitment was tested by Boyden chamber chemotaxis assays. Unloaded liposomes alone stimulated significant progenitor cell recruitment, while DDM loaded liposomes further promoted chemotactic recruitment in a dose dependent manner. DDM liposomes promoted the upregulation of 'osteodentine' markers osteocalcin and RUNX2 in DPSCs after 9 days of treatment, determined by Real Time quantitative PCR. Furthermore, Alizarin Red S staining showed that unloaded liposomes alone induced biomineralisation of DPSCs and DDM liposomes further increased the amount of mineralization observed. DDM liposomes were more effective than free DDM (10 μg/mL) at activating recruitment and osteogenic differentiation of DPSC, which are key events in the endogenous repair of the dentine-pulp complex. The study has highlighted the therapeutic potential of bioactive DDM liposomes in activating Dental Tissue repair in vitro, suggesting that liposomal delivery from biomaterials could be a valuable tool for reparative dentistry and hard Tissue engineering applications.

G Melling - One of the best experts on this subject based on the ideXlab platform.

  • liposomal delivery of demineralized dentin matrix for Dental Tissue Regeneration
    Tissue Engineering Part A, 2018
    Co-Authors: G Melling, John S Colombo, Steven J Avery, Wayne Nishio Ayre, Samuel Lewin Evans, Rachel J Waddington, Alastair James Sloan
    Abstract:

    Current Dental restorations have short longevity, consequently there is a need for novel Tissue engineering strategies that aim to regenerate the dentine-pulp complex. Dentine matrix contains a myriad of bioactive growth factors and extracellular matrix proteins associated with the recruitment, proliferation and differentiation of Dental pulp progenitor cells. Here, we show that demineralised dentine matrix (DDM), from non-carious dentine, can be encapsulated into liposomes for delivery to Dental Tissue to promote Regeneration. Liposomes were formulated to encapsulate 0 - 100 μg/mL DDM, lysed with Triton X and used in VEGF and TGF-β1 ELISAs to quantify release. The encapsulation efficiency was calculated to be 25.9% and 28.8% (VEGF/TGF-β1) for 50 μg/mL DDM liposomes and 39% and 146.7% (VEGF/TGF-β1) for 100 μg/mL DDM liposomes. All liposome formulations had no cytotoxic effects on a Dental pulp stem cell (DPSC) clone, as shown by MTT, Caspase 3/7 assays and cell counts. The ability of the liposomes to stimulate DPSC chemotactic recruitment was tested by Boyden chamber chemotaxis assays. Unloaded liposomes alone stimulated significant progenitor cell recruitment, while DDM loaded liposomes further promoted chemotactic recruitment in a dose dependent manner. DDM liposomes promoted the upregulation of 'osteodentine' markers osteocalcin and RUNX2 in DPSCs after 9 days of treatment, determined by Real Time quantitative PCR. Furthermore, Alizarin Red S staining showed that unloaded liposomes alone induced biomineralisation of DPSCs and DDM liposomes further increased the amount of mineralization observed. DDM liposomes were more effective than free DDM (10 μg/mL) at activating recruitment and osteogenic differentiation of DPSC, which are key events in the endogenous repair of the dentine-pulp complex. The study has highlighted the therapeutic potential of bioactive DDM liposomes in activating Dental Tissue repair in vitro, suggesting that liposomal delivery from biomaterials could be a valuable tool for reparative dentistry and hard Tissue engineering applications.

  • Liposomal Delivery of Demineralized Dentin Matrix for Dental Tissue Regeneration
    'Mary Ann Liebert Inc', 2018
    Co-Authors: G Melling, Wayne Nishio Ayre, Js Colombo, Sj Avery, Sl Evans, Rj Waddington, Aj Sloan
    Abstract:

    Current Dental restorations have short longevity, and consequently, there is a need for novel Tissue engineering strategies that aim to regenerate the dentin-pulp complex. Dentin matrix contains a myriad of bioactive growth factors and extracellular matrix proteins associated with the recruitment, proliferation, and differentiation of Dental pulp progenitor cells. In this study, we show that demineralized dentin matrix (DDM), from noncarious dentine, can be encapsulated into liposomes for delivery to Dental Tissue to promote Regeneration. Liposomes were formulated to encapsulate 0-100 μg/mL DDM, lysed with Triton X, and used in vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) enzyme-linked immunosorbent assays to quantify release. The encapsulation efficiencies were calculated to be 25.9% and 28.8% (VEGF/TGF-β1) for 50 μg/mL DDM liposomes and 39% and 146.7% (VEGF/TGF-β1) for 100 μg/mL DDM liposomes. All liposome formulations had no cytotoxic effects on a Dental pulp stem cell (DPSC) clone, as shown by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertrazolium bromide), Caspase 3/7 assays, and cell counts. The ability of the liposomes to stimulate DPSC chemotactic recruitment was tested by Boyden chamber chemotaxis assays. Unloaded liposomes alone stimulated significant progenitor cell recruitment, while DDM-loaded liposomes further promoted chemotactic recruitment in a dose-dependent manner. DDM liposomes promoted the upregulation of "osteodentin" markers osteocalcin and RUNX2 (Runt-related transcription factor 2) in DPSCs after 9 days of treatment, determined by real-time quantitative PCR. Furthermore, Alizarin Red S staining showed that unloaded liposomes alone induced biomineralization of DPSCs, and DDM liposomes further increased the amount of mineralization observed. DDM liposomes were more effective than free DDM (10 μg/mL) at activating recruitment and osteogenic differentiation of DPSC, which are key events in the endogenous repair of the dentin-pulp complex. The study has highlighted the therapeutic potential of bioactive DDM liposomes in activating Dental Tissue repair in vitro, suggesting that liposomal delivery from biomaterials could be a valuable tool for reparative dentistry and hard-Tissue engineering applications

Wayne Nishio Ayre - One of the best experts on this subject based on the ideXlab platform.

  • liposomal delivery of demineralized dentin matrix for Dental Tissue Regeneration
    Tissue Engineering Part A, 2018
    Co-Authors: G Melling, John S Colombo, Steven J Avery, Wayne Nishio Ayre, Samuel Lewin Evans, Rachel J Waddington, Alastair James Sloan
    Abstract:

    Current Dental restorations have short longevity, consequently there is a need for novel Tissue engineering strategies that aim to regenerate the dentine-pulp complex. Dentine matrix contains a myriad of bioactive growth factors and extracellular matrix proteins associated with the recruitment, proliferation and differentiation of Dental pulp progenitor cells. Here, we show that demineralised dentine matrix (DDM), from non-carious dentine, can be encapsulated into liposomes for delivery to Dental Tissue to promote Regeneration. Liposomes were formulated to encapsulate 0 - 100 μg/mL DDM, lysed with Triton X and used in VEGF and TGF-β1 ELISAs to quantify release. The encapsulation efficiency was calculated to be 25.9% and 28.8% (VEGF/TGF-β1) for 50 μg/mL DDM liposomes and 39% and 146.7% (VEGF/TGF-β1) for 100 μg/mL DDM liposomes. All liposome formulations had no cytotoxic effects on a Dental pulp stem cell (DPSC) clone, as shown by MTT, Caspase 3/7 assays and cell counts. The ability of the liposomes to stimulate DPSC chemotactic recruitment was tested by Boyden chamber chemotaxis assays. Unloaded liposomes alone stimulated significant progenitor cell recruitment, while DDM loaded liposomes further promoted chemotactic recruitment in a dose dependent manner. DDM liposomes promoted the upregulation of 'osteodentine' markers osteocalcin and RUNX2 in DPSCs after 9 days of treatment, determined by Real Time quantitative PCR. Furthermore, Alizarin Red S staining showed that unloaded liposomes alone induced biomineralisation of DPSCs and DDM liposomes further increased the amount of mineralization observed. DDM liposomes were more effective than free DDM (10 μg/mL) at activating recruitment and osteogenic differentiation of DPSC, which are key events in the endogenous repair of the dentine-pulp complex. The study has highlighted the therapeutic potential of bioactive DDM liposomes in activating Dental Tissue repair in vitro, suggesting that liposomal delivery from biomaterials could be a valuable tool for reparative dentistry and hard Tissue engineering applications.

  • Liposomal Delivery of Demineralized Dentin Matrix for Dental Tissue Regeneration
    'Mary Ann Liebert Inc', 2018
    Co-Authors: G Melling, Wayne Nishio Ayre, Js Colombo, Sj Avery, Sl Evans, Rj Waddington, Aj Sloan
    Abstract:

    Current Dental restorations have short longevity, and consequently, there is a need for novel Tissue engineering strategies that aim to regenerate the dentin-pulp complex. Dentin matrix contains a myriad of bioactive growth factors and extracellular matrix proteins associated with the recruitment, proliferation, and differentiation of Dental pulp progenitor cells. In this study, we show that demineralized dentin matrix (DDM), from noncarious dentine, can be encapsulated into liposomes for delivery to Dental Tissue to promote Regeneration. Liposomes were formulated to encapsulate 0-100 μg/mL DDM, lysed with Triton X, and used in vascular endothelial growth factor (VEGF) and transforming growth factor-β1 (TGF-β1) enzyme-linked immunosorbent assays to quantify release. The encapsulation efficiencies were calculated to be 25.9% and 28.8% (VEGF/TGF-β1) for 50 μg/mL DDM liposomes and 39% and 146.7% (VEGF/TGF-β1) for 100 μg/mL DDM liposomes. All liposome formulations had no cytotoxic effects on a Dental pulp stem cell (DPSC) clone, as shown by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltertrazolium bromide), Caspase 3/7 assays, and cell counts. The ability of the liposomes to stimulate DPSC chemotactic recruitment was tested by Boyden chamber chemotaxis assays. Unloaded liposomes alone stimulated significant progenitor cell recruitment, while DDM-loaded liposomes further promoted chemotactic recruitment in a dose-dependent manner. DDM liposomes promoted the upregulation of "osteodentin" markers osteocalcin and RUNX2 (Runt-related transcription factor 2) in DPSCs after 9 days of treatment, determined by real-time quantitative PCR. Furthermore, Alizarin Red S staining showed that unloaded liposomes alone induced biomineralization of DPSCs, and DDM liposomes further increased the amount of mineralization observed. DDM liposomes were more effective than free DDM (10 μg/mL) at activating recruitment and osteogenic differentiation of DPSC, which are key events in the endogenous repair of the dentin-pulp complex. The study has highlighted the therapeutic potential of bioactive DDM liposomes in activating Dental Tissue repair in vitro, suggesting that liposomal delivery from biomaterials could be a valuable tool for reparative dentistry and hard-Tissue engineering applications

Yu Cao - One of the best experts on this subject based on the ideXlab platform.

  • WIF1 enhanced dentinogenic differentiation in stem cells from apical papilla
    BMC Oral Health, 2019
    Co-Authors: Haifeng Wang, Yu Cao
    Abstract:

    Odontogenic mesenchymal stem cells (MSCs) isolated from tooth Tissues are a reliable resource that can be utilized for Dental Tissue Regeneration. Exploration of the mechanisms underlying the regulation of their differentiation may be helpful for investigating potential clinical applications. The stem cell niche plays an important role in maintaining cell functioning. Previous studies found that Wnt inhibitory factor 1 (WIF1) is more highly expressed in apical papilla Tissues than in stem cells from apical papilla (SCAPs) using microarray analysis. However, the function of WIF1 in SCAPs remains unclear. In the present study, we investigated the function of WIF1 during dentinogenic differentiation in SCAPs. A retrovirus containing HA-WIF1 was used to overexpress WIF1 in SCAPs. Using Western blot analysis, we verified the expression of HA-WIF1. Alkaline phosphatase (ALP) activity assays, Alizarin Red staining and quantitative calcium analysis were performed to investigate the in vitro potential for dentinogenic differentiation in SCAPs. The expression of dentinogenesis-associated genes DSPP, DMP1, Runx2 and OSX were assayed using real-time RT-PCR. Transplantation experiments were used to measure dentinogenesis potential in vivo. The real time RT-PCR results showed that WIF1 was more highly expressed in apical papilla Tissues than in SCAPs, and its expression was increased during the process of dentinogenic differentiation. Overexpression of WIF1 enhanced ALP activity and mineralization in vitro, as well as the expression of DSPP, DMP1 and OSX in SCAPs. Moreover, in vivo transplantation experiments revealed that dentinogenesis in SCAPs was enhanced by WIF1 overexpression. These results suggest that WIF1 may enhance dentinogenic differentiation potential in Dental MSCs via its regulation of OSX and identified potential target genes that could be useful for improving Dental Tissue Regeneration.

  • WIF1 enhanced dentinogenic differentiation in stem cells from apical papilla
    'Springer Science and Business Media LLC', 2019
    Co-Authors: Haifeng Wang, Yu Cao
    Abstract:

    Abstract Background Odontogenic mesenchymal stem cells (MSCs) isolated from tooth Tissues are a reliable resource that can be utilized for Dental Tissue Regeneration. Exploration of the mechanisms underlying the regulation of their differentiation may be helpful for investigating potential clinical applications. The stem cell niche plays an important role in maintaining cell functioning. Previous studies found that Wnt inhibitory factor 1 (WIF1) is more highly expressed in apical papilla Tissues than in stem cells from apical papilla (SCAPs) using microarray analysis. However, the function of WIF1 in SCAPs remains unclear. In the present study, we investigated the function of WIF1 during dentinogenic differentiation in SCAPs. Methods A retrovirus containing HA-WIF1 was used to overexpress WIF1 in SCAPs. Using Western blot analysis, we verified the expression of HA-WIF1. Alkaline phosphatase (ALP) activity assays, Alizarin Red staining and quantitative calcium analysis were performed to investigate the in vitro potential for dentinogenic differentiation in SCAPs. The expression of dentinogenesis-associated genes DSPP, DMP1, Runx2 and OSX were assayed using real-time RT-PCR. Transplantation experiments were used to measure dentinogenesis potential in vivo. Results The real time RT-PCR results showed that WIF1 was more highly expressed in apical papilla Tissues than in SCAPs, and its expression was increased during the process of dentinogenic differentiation. Overexpression of WIF1 enhanced ALP activity and mineralization in vitro, as well as the expression of DSPP, DMP1 and OSX in SCAPs. Moreover, in vivo transplantation experiments revealed that dentinogenesis in SCAPs was enhanced by WIF1 overexpression. Conclusion These results suggest that WIF1 may enhance dentinogenic differentiation potential in Dental MSCs via its regulation of OSX and identified potential target genes that could be useful for improving Dental Tissue Regeneration

  • SFRP2 enhances the osteogenic differentiation of apical papilla stem cells by antagonizing the canonical WNT pathway.
    Cellular & Molecular Biology Letters, 2017
    Co-Authors: Luyuan Jin, Shu Diao, Xiao Lin, Liping Wang, Yu Cao, Jinsong Wang, Xiaomeng Lian
    Abstract:

    Exploring the molecular mechanisms underlying directed differentiation is helpful in the development of clinical applications of mesenchymal stem cells (MSCs). Our previous study on Dental Tissue-derived MSCs demonstrated that secreted frizzled-related protein 2 (SFRP2), a Wnt inhibitor, could enhance osteogenic differentiation in stem cells from the apical papilla (SCAPs). However, how SFRP2 promotes osteogenic differentiation of Dental Tissue-derived MSCs remains unclear. In this study, we used SCAPs to investigate the underlying mechanisms. SCAPs were isolated from the apical papilla of immature third molars. Western blot and real-time RT-PCR were applied to detect the expression of β-catenin and Wnt target genes. Alizarin Red staining, quantitative calcium analysis, transwell cultures and in vivo transplantation experiments were used to study the osteogenic differentiation potential of SCAPs. SFRP2 inhibited canonical Wnt signaling by enhancing phosphorylation and decreasing the expression of nuclear β-catenin in vitro and in vivo. In addition, the target genes of the Wnt signaling pathway, AXIN2 (axin-related protein 2) and MMP7 (matrix metalloproteinase-7), were downregulated by SFRP2. WNT1 inhibited the osteogenic differentiation potential of SCAPs. SFRP2 could rescue this WNT1-impaired osteogenic differentiation potential. The results suggest that SFRP2 could bind to locally present Wnt ligands and alter the balance of intracellular Wnt signaling to antagonize the canonical Wnt pathway in SCAPs. This elucidates the molecular mechanism underlying the SFRP2-mediated directed differentiation of SCAPs and indicates potential target genes for improving Dental Tissue Regeneration.

Xiaohua Liu - One of the best experts on this subject based on the ideXlab platform.

  • magnesium containing nanostructured hybrid scaffolds for enhanced dentin Regeneration
    Tissue Engineering Part A, 2014
    Co-Authors: Junjun Jing, Yong Jiang, Robert J Taylor, Jian Q Feng, Benjamin Geiger, Xiaohua Liu
    Abstract:

    Dental caries is one of the most prevalent chronic diseases in the United States, affecting 92% of adults aged 20–64 years. Scaffold-based Tissue engineering represents a promising strategy to replace damaged Dental structures and restore their biological functions. Current single-component scaffolding materials used for Dental Tissue Regeneration, however, cannot provide the proper microenvironment for Dental stem/progenitor cell adhesion, proliferation, and differentiation; new biomimetic hybrid scaffolds are needed to promote better Dental Tissue formation. In this work, we developed a biomimetic approach to prepare three-dimensional (3D) nanofibrous gelatin/magnesium phosphate (NF-gelatin/MgP) hybrid scaffolds. These scaffolds not only mimic the nanostructured architecture and the chemical composition of natural dentin matrices but also constantly present favorable chemical signals (Mg ions) to Dental pulp stem cells (DPSCs), thus providing a desirable microenvironment to facilitate DPSC proliferation, differentiation, and biomineralization. Synthesized hybrid NF-gelatin/MgP possesses natural extracellular matrix (ECM)-like architecture, high porosity, high pore interconnectivity, well-defined pore size, and controlled Mg ion release from the scaffold. Adding MgP into NF-gelatin also increased the mechanical strength of the hybrid scaffold. The sustained release of Mg ions from the NF-gelatin/MgP (MgP=10% wt/wt) scaffold significantly enhanced the proliferation, differentiation, and biomineralization of human DPSCs in vitro. The alkaline phosphatase (ALP) activity and the gene expressions for odontogenic differentiation (collagen I [Col I], ALP, osteocalcin [OCN], dentin sialophosphoprotein [DSPP], and dentin matrix protein 1 [DMP1]) were all significantly higher (p<0.05) in the NF-gelatin/MgP group than in the NF-gelatin group. Those results were further confirmed by hematoxylin and eosin (H&E) and von Kossa staining, as shown by greater ECM secretion and mineral deposition in the hybrid scaffold. Consistent with the in vitro study, the DPSCs/NF-gelatin/MgP constructs produced greater ECM deposition, hard Tissue formation, and expression of marker proteins (DSPP, DMP1, Col I) for odontogenic differentiation than did the DPSCs/NF-gelatin after 5 weeks of ectopic implantation in rude mice. The controlled release of metallic ions from biomimetic nanostructured hybrid scaffolds, therefore, is a promising approach to enhancing the biological capability of the scaffolds for Dental Tissue Regeneration.