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Dental follicle stem cells in bone regeneration on titanium implants.

Lucaciu O, Soriţău O, Gheban D, Ciuca DR, Virtic O, Vulpoi A, Dirzu N, Câmpian R, Băciuţ G, Popa C, Simon S, Berce P, Băciuţ M, Crisan B - BMC Biotechnol. (2015)

Bottom Line: Differentiation into bone cells was induced in presence or absence of BMP-2 and TGFβ1.Additional BMP-2 in the medium did not allow DF stem cells to develop a more mature phenotype, leaving them in a pre-osteogenic stage.Long term culturing (70 days) on titanium surfaces of DF stem cells in standard medium without soluble osteogenic inducers, indicated that HA coating is more favorable, with the acquisition of a more mature osteoblastic phenotype as shown by immunocytochemical staining.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Rehabilitation, "Iuliu Haţieganu" University of Medicine and Pharmacy Cluj-Napoca, 15 Victor Babeș Street, 400012, Cluj-Napoca, Cluj, Romania. ondineluc@yahoo.com.

ABSTRACT

Background: We aimed to demonstrate that DF stem cells from impacted molars and canines can be used to improve bone regeneration on titanium implants surfaces. This study highlights the presence of stem cells in DF, their potential to adhere and differentiate into osteoblasts on different types of titanium surfaces.

Results: Isolated cells from the harvested DF tissue from impacted canine/molars, expressed stem cells markers. Differentiation into bone cells was induced in presence or absence of BMP-2 and TGFβ1. The presence of growth factors until 28 days in medium maintained the cells in an earlier stage of differentiation with a lower level of specific bone proteins and a higher expression of alkaline phosphatase (ALP). Influence of titanium implants with different bioactive coatings, hydroxyapatite (TiHA) and with silicatitanate (TiSiO2), and porous Ti6Al7Nb implants as control (TiCtrl), was studied in terms of cell adhesion and viability. Ti HA implants proved to be more favorable for adhesion and proliferation of DF stem cells in first days of cultivation. The influence of titanium coatings and osteogenic differentiation mediums with or without growth factors were evaluated. Additional BMP-2 in the medium did not allow DF stem cells to develop a more mature phenotype, leaving them in a pre-osteogenic stage. The best sustained mineralization process evaluated by immuno-cytochemical staining, scanning electron microscopy and Ca(2+) quantification was observed for TiHA implants with a higher expression of ALP, collagen and Ca(2+) deposition. Long term culturing (70 days) on titanium surfaces of DF stem cells in standard medium without soluble osteogenic inducers, indicated that HA coating is more favorable, with the acquisition of a more mature osteoblastic phenotype as shown by immunocytochemical staining. These findings demonstrated that even in absence of exogenous osteogenic factors, TiHA implants and in a lesser extent TiCtrl and TiSiO2 implants can induce and sustain osteogenic differentiation of DF stem cells, by their chemical and topographical properties.

Conclusions: Our research demonstrated that DF stem cells have a spontaneous tendency for osteogenic differentiation and can be used for improving bone regeneration on titanium implants surfaces.

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Related in: MedlinePlus

Immunocytochemical staining for characteristic stem cells markers, nuclei were counterstained with DAPI. (a) strong expression for CD 44 FITC and weak positivity for CD 105 PE; (b) negative staining for CD 34 FITC and CD 117 PE; (c) strong positivity for CD 90 FITC, weak expression of CD 49 PE; (d) strong positivity for early embryonic antigen SSEA-4 FITC; (e) the self-renewal embryonic proteins Oct3/4 FITC strong expression and (f) Nanog FITC positive in some cells (magnification ×400)
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Fig4: Immunocytochemical staining for characteristic stem cells markers, nuclei were counterstained with DAPI. (a) strong expression for CD 44 FITC and weak positivity for CD 105 PE; (b) negative staining for CD 34 FITC and CD 117 PE; (c) strong positivity for CD 90 FITC, weak expression of CD 49 PE; (d) strong positivity for early embryonic antigen SSEA-4 FITC; (e) the self-renewal embryonic proteins Oct3/4 FITC strong expression and (f) Nanog FITC positive in some cells (magnification ×400)

Mentions: Immunocytochemical staining performed for isolated cells at third passage has shown strong positivity for specific markers of mesenchymal stem cells such as: SSEA-4, Oct3/4, Nanog, CD44, D90, and weak positivity for CD105 and CD49e. Cells did not express CD 117 and CD34 (Fig. 4).Fig. 4


Dental follicle stem cells in bone regeneration on titanium implants.

Lucaciu O, Soriţău O, Gheban D, Ciuca DR, Virtic O, Vulpoi A, Dirzu N, Câmpian R, Băciuţ G, Popa C, Simon S, Berce P, Băciuţ M, Crisan B - BMC Biotechnol. (2015)

Immunocytochemical staining for characteristic stem cells markers, nuclei were counterstained with DAPI. (a) strong expression for CD 44 FITC and weak positivity for CD 105 PE; (b) negative staining for CD 34 FITC and CD 117 PE; (c) strong positivity for CD 90 FITC, weak expression of CD 49 PE; (d) strong positivity for early embryonic antigen SSEA-4 FITC; (e) the self-renewal embryonic proteins Oct3/4 FITC strong expression and (f) Nanog FITC positive in some cells (magnification ×400)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4697321&req=5

Fig4: Immunocytochemical staining for characteristic stem cells markers, nuclei were counterstained with DAPI. (a) strong expression for CD 44 FITC and weak positivity for CD 105 PE; (b) negative staining for CD 34 FITC and CD 117 PE; (c) strong positivity for CD 90 FITC, weak expression of CD 49 PE; (d) strong positivity for early embryonic antigen SSEA-4 FITC; (e) the self-renewal embryonic proteins Oct3/4 FITC strong expression and (f) Nanog FITC positive in some cells (magnification ×400)
Mentions: Immunocytochemical staining performed for isolated cells at third passage has shown strong positivity for specific markers of mesenchymal stem cells such as: SSEA-4, Oct3/4, Nanog, CD44, D90, and weak positivity for CD105 and CD49e. Cells did not express CD 117 and CD34 (Fig. 4).Fig. 4

Bottom Line: Differentiation into bone cells was induced in presence or absence of BMP-2 and TGFβ1.Additional BMP-2 in the medium did not allow DF stem cells to develop a more mature phenotype, leaving them in a pre-osteogenic stage.Long term culturing (70 days) on titanium surfaces of DF stem cells in standard medium without soluble osteogenic inducers, indicated that HA coating is more favorable, with the acquisition of a more mature osteoblastic phenotype as shown by immunocytochemical staining.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Rehabilitation, "Iuliu Haţieganu" University of Medicine and Pharmacy Cluj-Napoca, 15 Victor Babeș Street, 400012, Cluj-Napoca, Cluj, Romania. ondineluc@yahoo.com.

ABSTRACT

Background: We aimed to demonstrate that DF stem cells from impacted molars and canines can be used to improve bone regeneration on titanium implants surfaces. This study highlights the presence of stem cells in DF, their potential to adhere and differentiate into osteoblasts on different types of titanium surfaces.

Results: Isolated cells from the harvested DF tissue from impacted canine/molars, expressed stem cells markers. Differentiation into bone cells was induced in presence or absence of BMP-2 and TGFβ1. The presence of growth factors until 28 days in medium maintained the cells in an earlier stage of differentiation with a lower level of specific bone proteins and a higher expression of alkaline phosphatase (ALP). Influence of titanium implants with different bioactive coatings, hydroxyapatite (TiHA) and with silicatitanate (TiSiO2), and porous Ti6Al7Nb implants as control (TiCtrl), was studied in terms of cell adhesion and viability. Ti HA implants proved to be more favorable for adhesion and proliferation of DF stem cells in first days of cultivation. The influence of titanium coatings and osteogenic differentiation mediums with or without growth factors were evaluated. Additional BMP-2 in the medium did not allow DF stem cells to develop a more mature phenotype, leaving them in a pre-osteogenic stage. The best sustained mineralization process evaluated by immuno-cytochemical staining, scanning electron microscopy and Ca(2+) quantification was observed for TiHA implants with a higher expression of ALP, collagen and Ca(2+) deposition. Long term culturing (70 days) on titanium surfaces of DF stem cells in standard medium without soluble osteogenic inducers, indicated that HA coating is more favorable, with the acquisition of a more mature osteoblastic phenotype as shown by immunocytochemical staining. These findings demonstrated that even in absence of exogenous osteogenic factors, TiHA implants and in a lesser extent TiCtrl and TiSiO2 implants can induce and sustain osteogenic differentiation of DF stem cells, by their chemical and topographical properties.

Conclusions: Our research demonstrated that DF stem cells have a spontaneous tendency for osteogenic differentiation and can be used for improving bone regeneration on titanium implants surfaces.

Show MeSH
Related in: MedlinePlus