Human dental pulp stem cells produce mineralized matrix in 2D and 3D cultures

Riccio, Massimo; Resca, Elisa; Maraldi, Tullia; Pisciotta, Alessandra; Ferrari, Adriano; Bruzzesi, Giacomo; Pol, Anto De

doi:10.4081/ejh.2010.e46

Cited by 62 publications

(48 citation statements)

References 37 publications

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“…1,14,16,18-21 Odontoblasts are post-mitotic cells unable to proliferate, which makes their cultivation by traditional cell culture systems unsuccessful. Some authors have suggested culture methods for human odontoblasts based on the dissected crowns of molars, in which the pulp was removed and the odontoblast layer, attached to the walls of the pulp chamber, was cultured in cell medium 18 .…”

Section: Discussionmentioning

confidence: 99%

“…The TGF-β1 is one of the most abundant cytokines found in bone matrix and is a potent stimulator of tissue regeneration and a fibrogenic mediator, 15 and its ability to promote fibroblastic differentiation has already been investigated in other systems. 16,17 We chose TGF-beta1 treatment in order to significantly increase the differentiative potential of pulp cells and to accelerate the regeneration of functional dentin. Alizarin red staining, ultrastructural morphological analysis by electron microscopy, and biochemical assays of type 1 collagen, and DMP1 and DSP expressions were carried out to demonstrate the odontoblast phenotype.…”

Section: Introductionmentioning

confidence: 99%

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In vitro reparative dentin: a biochemical and morphological study

et al. 2013

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In this study, starting from human dental pulp cells cultured in vitro, we simulated reparative dentinogenesis using a medium supplemented with different odontogenic inductors. The differentiation of dental pulp cells in odontoblast-like cells was evaluated by means of staining, and ultramorphological, biochemical and biomolecular methods. Alizarin red staining showed mineral deposition while transmission electron microscopy revealed a synthesis of extracellular matrix fibers during the differentiation process. Biochemical assays demonstrated that the differentiated phenotype expressed odontoblast markers, such as Dentin Matrix Protein 1 (DMP1) and Dentin Sialoprotein (DSP), as well as type I collagen. Quantitative data regarding the mRNA expression of DMP1, DSP and type I collagen were obtained by Real Time PCR. Immunofluorescence data demonstrated the various localizations of DSP and DMP1 during odontoblast differentiation. Based on our results, we obtained odontoblast-like cells which simulated the reparative dentin processes in order to better investigate the mechanism of odontoblast differentiation, and dentin extracellular matrix deposition and mineralization.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In vitro reparative dentin: a biochemical and morphological study

et al. 2013

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“…All procedures regarding collection of human samples were approved by the Provincial Ethics Committee of Santa Maria Nuova Hospital of Reggio Emilia. hDPSCs were isolated from dental pulp, as described in a previous study [8]. Briefly cells obtained from dental pulp were plated at clonal density (1.6 cells/cm 2 ) and after 6 days of culture, nodules originated by single cells were isolated from the culture plate, re-plated and expanded.…”

Section: Methodsmentioning

confidence: 99%

“…After 8 hours from cell seeding, 2 ml of the two osteogenic media were added to each sample. hDPSCs/collagen constructs were pre-differentiated for 10 days before in vivo implantation according to previous data [8], [10].…”

Section: Methodsmentioning

confidence: 99%

Human Serum Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells In Vitro and In Vivo

et al. 2012

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Human dental pulp is a promising alternative source of stem cells for cell-based tissue engineering in regenerative medicine, for the easily recruitment with low invasivity for the patient and for the self-renewal and differentiation potential of cells. So far, in vitro culture of mesenchymal stem cells is usually based on supplementing culture and differentiation media with foetal calf serum (FCS). FCS is known to contain a great quantity of growth factors, and thus to promote cell attachment on plastic surface as well as expansion and differentiation. Nevertheless, FCS as an animal origin supplement may represent a potential means for disease transmission besides leading to a xenogenic immune response. Therefore, a significant interest is focused on investigating alternative supplements, in order to obtain a sufficient cell number for clinical application, avoiding the inconvenients of FCS use. In our study we have demonstrated that human serum (HS) is a suitable alternative to FCS, indeed its addition to culture medium induces a high hDPSCs proliferation rate and improves the in vitro osteogenic differentiation. Furthermore, hDPSCs-collagen constructs, pre-differentiated with HS-medium in vitro for 10 days, when implanted in immunocompromised rats, are able to restore critical size parietal bone defects. Therefore these data indicate that HS is a valid substitute for FCS to culture and differentiate in vitro hDPSCs in order to obtain a successful bone regeneration in vivo.

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“…The confocal serial sections were processed with ImageJ software to obtain three-dimensional projections, as previously described [41]. The image rendering was performed by Adobe Photoshop software.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of Nuclear Nox4 Activity by Plumbagin: Effect on Proliferative Capacity in Human Amniotic Stem Cells

Guida

Maraldi

Resca

et al. 2013

Oxidative Medicine and Cellular Longevity

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Human amniotic fluid stem cells (AFSC) with multilineage differentiation potential are novel source for cell therapy. However, in vitro expansion leads to senescence affecting differentiation and proliferative capacities. Reactive oxygen species (ROS) have been involved in the regulation of stem cell pluripotency, proliferation, and differentiation. Redox-regulated signal transduction is coordinated by spatially controlled production of ROS within subcellular compartments. NAD(P)H oxidase family, in particular Nox4, has been known to produce ROS in the nucleus; however, the mechanisms and the meaning of this function remain largely unknown. In the present study, we show that Nox4 nuclear expression (nNox4) increases during culture passages up to cell cycle arrest and the serum starvation causes the same effect. With the decrease of Nox4 activity, obtained with plumbagin, a decline of nuclear ROS production and of DNA damage occurs. Moreover, plumbagin exposure reduces the binding between nNox4 and nucleoskeleton components, as Matrin 3. The same effect was observed also for the binding with phospho-ERK, although nuclear ERK and P-ERK are unchanged. Taken together, we suggest that nNox4 regulation may have important pathophysiologic effects in stem cell proliferation through modulation of nuclear signaling and DNA damage.

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Human dental pulp stem cells produce mineralized matrix in 2D and 3D cultures

Cited by 62 publications

References 37 publications

In vitro reparative dentin: a biochemical and morphological study

In vitro reparative dentin: a biochemical and morphological study

Human Serum Promotes Osteogenic Differentiation of Human Dental Pulp Stem Cells In Vitro and In Vivo

Inhibition of Nuclear Nox4 Activity by Plumbagin: Effect on Proliferative Capacity in Human Amniotic Stem Cells

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