Expression of BSP-GFPtpz Transgene during Osteogenesis and Reparative Dentinogenesis

Vijaykumar, Anushree; Dyrkacz, P; Vidovic-Zdrilic, I; Maye, Peter; Mina, M.

doi:10.1177/0022034519885089

Cited by 24 publications

(26 citation statements)

References 42 publications

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“…ALP is often detected in the early stages of bone retraction and thus is used to measure levels of early osteogenesis. On the other hand, BSP is found in mineralized tissues such as dentin, but the exact mechanism and involvement of BSP in bone regeneration is still being investigated [ 50 ]. OC is secreted by osteogenic-related cells during osteogenesis and is known to act on calcium ions to bring about mineralization.…”

Section: Resultsmentioning

confidence: 99%

Biofabrication of Gingival Fibroblast Cell-Laden Collagen/Strontium-Doped Calcium Silicate 3D-Printed Bi-Layered Scaffold for Osteoporotic Periodontal Regeneration

et al. 2021

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Periodontal disease is a chronic disease that can lead to lose teeth and even tooth loss if left untreated. Osteoporosis and periodontal disease share similar characteristics and associated factors. Current regenerative techniques for periodontal diseases are ineffective in restoring complete function and structural integrity of periodontium due to unwanted migration of cells. In this study, we applied the concept of guided tissue regeneration (GTR) and 3D fabricated gingival fibroblast cell-laden collagen/strontium-doped calcium silicate (SrCS) bi-layer scaffold for periodontal regeneration. The results revealed that the bioactive SrCS had a hydroxyapatite formation on its surface after 14 days of immersion and that SrCS could release Sr and Si ions even after 6 months of immersion. In addition, in vitro results showed that the bi-layer scaffold enhanced secretion of FGF-2, BMP-2, and VEGF from human gingival fibroblasts and increased secretion of osteogenic-related proteins ALP, BSP, and OC from WJMSCs. In vivo studies using animal osteoporotic models showed that the 3D-printed cell-laden collagen/SrCS bi-layer scaffold was able to enhance osteoporotic bone regeneration, as seen from the increased Tb.Th and BV/TV ratio and the histological stains. In conclusion, it can be seen that the bi-layer scaffolds enhanced osteogenesis and further showed that guided periodontal regeneration could be achieved using collagen/SrCS scaffolds, thus making it a potential candidate for future clinical applications.

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Section: Resultsmentioning

confidence: 99%

Biofabrication of Gingival Fibroblast Cell-Laden Collagen/Strontium-Doped Calcium Silicate 3D-Printed Bi-Layered Scaffold for Osteoporotic Periodontal Regeneration

et al. 2021

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“…All experimental protocols involving animal tissues in the present study were approved by the Institutional Animal Care and use Committee of UConn Health Center. DSPP-Cerulean/DMP1-Cherry, BSP-GFPtpz, pOBCol2.3FP (referred to as 2.3-GFP), TCF/Lef:H2B-EGFP, and nontransgenic mice used in this study have been previously described (Balic, Aguila, Caimano, et al 2010; Ferrer-Vaquer et al 2010; Vijaykumar et al 2019, 2020). All mice were maintained in the CD1 background.…”

Section: Methodsmentioning

confidence: 99%

“…Medium was changed every other day. Mineralization in live primary cultures was examined by xylenol orange (XO) staining (Vijaykumar et al 2020). The mean fluorescence intensity of XO staining, DMP1-Cherry, and BSP-GFPtpz was measured using a microplate reader (Safire 2 ; Tecan) (Vijaykumar et al 2019, 2020).…”

Section: Methodsmentioning

confidence: 99%

“…DMP1-Cherry and DSPP-Cerulean transgenes are activated and expressed by functional and fully differentiated odontoblasts, respectively (Vijaykumar et al 2019). More recently, we showed that BSP-GFPtpz identifies cells in the osteogenic lineage and is excluded from cells in the dentinogenic lineage (Vijaykumar et al 2020). The lineage- and stage-specific activation of these transgenes provides a panel of reporters that can help identify and isolate cells in dentinogenic and osteogenic lineages at various stages of differentiation.…”

Section: Introductionmentioning

confidence: 99%

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Wnt/β-Catenin Signaling Promotes the Formation of Preodontoblasts In Vitro

2020

Self Cite

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Odontoblast differentiation is a complex and multistep process regulated by signaling pathways, including the Wnt/β-catenin signaling pathway. Both positive and negative effects of Wnt/β-catenin signaling on dentinogenesis have been reported, but the underlying mechanisms of these conflicting results are still unclear. To gain a better insight into the role of Wnt/β-catenin in dentinogenesis, we used dental pulp cells from a panel of transgenic mice, in which fluorescent protein expression identifies cells at different stages of odontoblast and osteoblast differentiation. Our results showed that exposure of pulp cells to WNT3a at various times and durations did not induce premature differentiation of odontoblasts. These treatments supported the survival of undifferentiated cells in dental pulp and promoted the formation of 2.3GFP+ preodontoblasts and their rapid transition into differentiated odontoblasts expressing DMP1-Cherry and DSPP-Cerulean transgenes. WNT3a also promoted osteogenesis in dental pulp cultures. These findings provide critical information for the development of improved treatments for vital pulp therapy and dentin regeneration.

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“…Although the effects of LiCl on reparative dentin and odontoblast differentiation has been established, much remains to be understood about the underlying cellular events leading to increased reparative dentinogenesis. Therefore, to gain a better understanding of the roles of LiCl on odontoblast and osteoblast differentiation, we used well-characterized dental pulp cultures from a series of previously characterized reporter mice (23)(24)(25) and examined the response of cells at various stages of differentiation to LiCl.…”

Section: Introductionmentioning

confidence: 99%

Lithium Chloride Exerts Differential Effects on Dentinogenesis and Osteogenesis in Primary Pulp Cultures

Vijaykumar¹,

Mina²

2021

Front. Dent. Med

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Wnt/β-catenin signaling is known to play essential roles in odontoblast differentiation and reparative dentin formation. Various Wnt activators including LiCl have been increasingly studied for their effectiveness to induce repair of the dentin-pulp complex. LiCl is a simple salt thought to activate Wnt/β-catenin signaling by inhibiting GSK3β. Previous in vitro and in vivo studies showed that LiCl increased odontoblast differentiation and enhanced reparative dentin formation. However, the underlying molecular and cellular mechanisms by which LiCl regulates odontoblast and osteoblast differentiation during reparative dentinogenesis are not well-understood. Our in vitro studies show that exposure of early dental pulp progenitors to LiCl increased the survival and the pool of αSMA+ progenitors, leading to enhanced odontoblast and osteoblast differentiation. The positive effects of LiCl in the differentiation of osteoblasts and odontoblasts from αSMA+ progenitors are mediated by Wnt/β-catenin signaling. Our results also showed that continuous and late exposure of dental pulp cells to LiCl increased the expression of odontoblast markers through Wnt/β-catenin signaling, and the number of odontoblasts expressing DMP1-Cherry and DSPP-Cerulean transgenes. However, unlike the early treatment, both continuous and late treatments decreased the expression of Bsp and the expression of BSP-GFPtpz transgene. These observations suggest that prolonged treatment with LiCl in more mature cells of the dental pulp has an inhibitory effect on osteoblast differentiation. The inhibitory effects of LiCl on osteogenesis and Bsp were not mediated through Wnt/β-catenin signaling. These observations suggest that the effects of LiCl, and GSK3β antagonists on reparative dentinogenesis involve multiple pathways and are not specific to Wnt/β-catenin signaling.

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Expression of BSP-GFPtpz Transgene during Osteogenesis and Reparative Dentinogenesis

Cited by 24 publications

References 42 publications

Biofabrication of Gingival Fibroblast Cell-Laden Collagen/Strontium-Doped Calcium Silicate 3D-Printed Bi-Layered Scaffold for Osteoporotic Periodontal Regeneration

Biofabrication of Gingival Fibroblast Cell-Laden Collagen/Strontium-Doped Calcium Silicate 3D-Printed Bi-Layered Scaffold for Osteoporotic Periodontal Regeneration

Wnt/β-Catenin Signaling Promotes the Formation of Preodontoblasts In Vitro

Lithium Chloride Exerts Differential Effects on Dentinogenesis and Osteogenesis in Primary Pulp Cultures

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