Artículos científicos año 2013

Abstract

Stem cells from human exfoliated deciduous teeth are a population of highly proliferative postnatal stem cells and have been characterized as multipotent stem cells. In this study we developed a fast and sensitive method for stem cells derived from human exfoliated deciduous teeth count, using a luminescent viability assay. We isolated stem cells from normal exfoliated deciduous teeth using collagenase/dispase digestions solutions. Separated stem cells were placed in opaque-walled multiwall plates in culture alpha Modified Eagle's Medium. For dental pulp stem cells quantitation we used a simple method for determining the number of viable cells based on ATP concentration. Cells attached to the bottom of the multiwall plates were counted with the luminescent assay and were cultured for mesenchymal markers expression. Moreover cells attached to the bottom of the multiwall plates were directed toward the osteogenic, adipogenic, lineages at the respective passages. Flow cytometry was used for immunophenotyping of cultured dental stem cells from exfoliated deciduous teeth. Cells that were counted with the luminescent assay, after culture formed fibroblastic morphology and were expressed the mesenchymal stem cell markers CD29, CD105, CD146, CD44. There was a correlation between the number of cells plated for culture and the number of mesenchymal stem cells after culture. Osteogenic and adipogenic differentiation of the cells counted with the luminescent assay was performed. The luminescent signal of viable mesenchymal dental stem cells isolated from dental pulp of exfoliated teeth represents an ideal method for mesenchymal stem cells count before culturing.

http://www.ncbi.nlm.nih.gov/pubmed/24379108. 

Abstract

Stem cells from human exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSCs) obtained from the dental pulp of human extracted tooth were cultured and characterized to confirm that these were mesenchymal stem cells. The proliferation rate was assessed using AlamarBlue® cell assay. The differentially expressed genes in SHED and DPSCs were identified using GeneFishing^TM technique. The proliferation rate of SHED (P < 0.05) was significantly higher than DPSCs while SHED had lesser multiplication rate and shorter population doubling time (0.01429, 60.57 hours) than DPSCs (0.00286, 472.43 hours). Two bands were highly expressed in SHED and three bands in DPSCs. Sequencing analysis revealed these to be TIMP Metallopeptidase Inhibitor 1 (TIMP1), and ribosomal protein s8, (RPS8) in SHED and collagen, type I, alpha 1, (COL1A1), follistatin-like 1 (FSTL1), lectin, galactoside-binding, soluble, 1, (LGALS1) in DPSCs.TIMP1is involved in degradation of the extracellular matrix, cell proliferation and anti-apoptotic function and RPS8 is involved as a rate-limiting factor in translational regulation; COL1A1 is involved in the resistance and elasticity of the tissues; FSTL1 is an autoantigen associated with rheumatoid arthritis; LGALS1 is involved in cell growth, differentiation, adhesion, RNA processing, apoptosis and malignant transformation. This along with further protein expression analysis holds promise in tissue engineering and regenerative medicine.

http://www.ncbi.nlm.nih.gov/pubmed/24375868. 

Abstract

Growing evidence show that human dental pulp stem cells (DPSCs) could provide a source of adult stem cells for the treatment of neurodegenerative pathologies. In this study, DPSCs were expanded and cultured with a protocol generally used for the culture of neural stem/progenitor cells.

Methodology:

DPSC cultures were established from third molars. The pulp tissue was enzymatically digested and cultured in serum-supplemented basal medium for 12 h. Adherent (ADH) and non-adherent (non-ADH) cell populations were separated according to their differential adhesion to plastic and then cultured in serum-free defined N₂ medium with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). Both ADH and non-ADH populations were analyzed by FACS and/or PCR.

Results:

FACS analysis of ADH-DPSCs revealed the expression of the mesenchymal cell marker CD90, the neuronal marker CD56, the transferrin receptor CD71, and the chemokine receptor CXCR3, whereas hematopoietic stem cellsmarkers CD45, CD133, and CD34 were not expressed. ADH-DPSCs expressed transcripts coding for the Nestin gene, whereas expression levels of genes coding for the neuronal markers β-III tubulin and NF-M, and the oligodendrocyte marker PLP-1 were donor dependent. ADH-DPSCs did not express the transcripts for GFAP, an astrocyte marker. Cells of the non-ADH population that grew as spheroids expressed Nestin, β-III tubulin, NF-M and PLP-1 transcripts. DPSCs that migrated out of the spheroids exhibited an odontoblast-like morphology and expressed a higher level of DSPP and osteocalcin transcripts than ADH-DPSCs.

Conclusion:

Collectively, these data indicate that human DPSCs can be expanded and cultured in serum-free supplemented medium with EGF and bFGF. ADH-DPSCs and non-ADH populations contained neuronal and/or oligodendrocyte progenitors at different stages of commitment and, interestingly, cells from spheroid structures seem to be more engaged into the odontoblastic lineage than the ADH-DPSCs.

http://www.ncbi.nlm.nih.gov/pubmed/24376422. 

Abstract

During recent years, attention has been given to the potential of therapeutic approaches using stem cells obtained from dental pulp tissue. The aim of this study, therefore, was to give an overview of the papers produced during the last 10 years that have described the use of stem cells obtained from human deciduous teeth in cell therapy or bioengineering. The PubMed database was investigated from January 2002 until July 2011 and the papers published during this period were analyzed according to criteria previously established, using the methodology of systematic review. The measurements were done using "stem cell" as the primary keyword, and "human deciduous teeth dental pulp cell" and "human exfoliated deciduous teeth" as the secondary keywords. Four hundred and seventy-five papers were found. The first screening resulted in 276 papers, from which 84 papers were selected. However, only 11 of them attained the aim proposed in our approach. There were few scientific studies related to direct therapeutic application using stem cells of human deciduous teeth and none of them had been applied to humans. However, the results indicated important and promising applications of the pulp stem-cells in cell therapy and bioengineering as demonstrated by studies in animal models of muscular dystrophy, Parkison's disease, and lupus erythematosus.

http://www.ncbi.nlm.nih.gov/pubmed/24130093. 

Abstract

Two types of dentition are generated in a human's lifetime: the primary dentition, followed by the permanent dentition. Undoubtedly, teeth are essential for speech and mastication in both dentitions, but it is becoming apparent that dental pulp also plays a role in harboring mesenchymal stem cells (MSCs). To date, three kinds of MSCs derived from dental pulp have been established: permanent tooth, primary tooth, and immature apical papilla. The dental pulp from primary teeth is considered a particularly good source of MSCs; it can be obtained from extracted primary teeth, of which humans have 20. The past decade has seen many reports of dental pulp-derived MSCs, and the field is becoming increasingly popular. The present article describes the characterization of dental pulp-derived MSCs from primary teeth. It also discusses future banking activity of primary teeth, because it is known that dental pulp-derived MSCs have similar potential to those derived from bone marrow. Methods with which to optimize the cryopreservation process should therefore be investigated, because banked dental pulp may provide a great resource in future regenerative medicine.

http://www.ncbi.nlm.nih.gov/pubmed/24278944. 

Abstract

AIMS:

Our aims were to characterize human mesenchymal stem cells isolated from various tissues by pluripotent stem cells gene expression profile.

METHODS:

Four strains of dental pulp stem cells (DP-MSCs) were isolated from dental pulp tissue fragments adhered to plastic tissue culture dishes. Mesenchymal stem cells derived from umbilical cord tissue (UBC-MSCs) were isolated with the same technique. Bone marrow derived mesenchymal stem cells (BM-MSCs) were isolated from nucleated cells of bone marrow obtained by density gradient centrifugation. Human mesenchymal stem cells from adipose tissue (AT-MSCs) were isolated by collagenase digestion. All kinds of MSCs used in this study were cultivated in low glucose DMEM containing 5% or human platelet extract. All stem cell manipulation was performed in GMP conditions. Expression of 15 pluripotent stem cells genes on the level of proteins was measured by Proteome Profiler Human Pluripotent Stem Cell Array. Induction of MSCs to in vitro differentiation to adipocytes, osteoblasts, chondroblasts was achieved by cultivation of cells in appropriate differentiation medium.

RESULTS:

All MSCs tested were phenotypically similar and of fibroblastoid morphology. DP-MSCs and UBC-MSCs were more proliferative than bone marrow BM-MSCs and AT-MSCs. Protein expression of 15 genes typical for pluripotent stem cells distinguished them into two groups. While the gene expression profiles of BM-MSC, AT-MSCs and UBC-MSCs were similar, DP-MSCS differed in relative gene expression on the level of their products in several genes.

CONCLUSIONS:

Dental pulp mesenchymal stem cells cultivated in vitro under the same conditions as MSCs from bone marrow, adipose tissue and umbilical cord tissue can be distinguished by pluripotent stem cell gene expression profile.

http://www.ncbi.nlm.nih.gov/pubmed/24145770.

Abstract

Mesenchymal stem cells can be obtained with ease from dental/oral tissue, making them an attractive source of autologous stem cells. They offer a biological solution for restoring damaged dental tissues such as vital pulp engineering, regeneration of periodontal ligament lost in periodontal disease, and for generation of complete or partial tooth structures to form biological implants. Dental mesenchymal stem cells share properties with mesenchymal stem cells from bone marrow and there is a considerable potential for these cells to be used in different stem-cell-based therapies, such as bone and muscle regeneration. In addition, their immunosuppressive-immunomodulatory properties make these cells a suitable source for treating immunodisorders like systematic lupus erythematosus. In addition, gingival tissue might also be a very good source of epithelial cells used in the treatment of severe ocular surface disorders. Being such an accessible source for different stem cells, the tooth and the attached gingival tissue (usually discarded in the clinics) represent an ideal source of autologous or allogeneic stem cells that can be used in the treatment of many clinical conditions in dentistry and medicine.

http://www.ncbi.nlm.nih.gov/pubmed/24113958.

Abstract

AIMS AND OBJECTIVES:

Isolation, characterization and differentiation of dental pulp stem cells (DPSCs) and stem cells from exfoliated human deciduous teeth (SHED).

METHODS:

The pulp tissue was digested in collagenase and cultured in DMEM Dulbecco's Modified Eagle's Media). The stem cells were identified and isolated. Surface characterization of cells was done with flow cytometer using surface markers. An immuno cytochemistry analysis was done. Differentiation potential was analyzed using various differentiation markers.

RESULTS:

Flow cytometry analyses for various CD markers showed similar results for both DPSCs and SHED. The cells showed positive expression for pluripotent, ectodermal and mesodermal markers. Cells differentiated into osteoblasts and adipocytes.

CONCLUSION:

The study demonstrated that stem cells existed in deciduous and permanent pulp tissue. The stem cells present in pulp tissue can be isolated, cultivated and expanded in vitro. Both DPSCs and SHED show almost a similar expression pattern profile for variety of antigens tested.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3778624/.

Abstract

Currently, much information is provided regarding the presence and the roles in tissue regeneration of stem cell niches residing in post-natal dental pulp. So far, three types of adult stem cells have been isolated from dental pulp. Correct evaluation of these cells is important in order to determine their potential use in clinical fields. The present study aims to review the origins and immunophenotype of these cells. The particularities of interstitial cells of the stem cell niches are also debated.

http://www.rjme.ro/RJME/resources/files/540313473478.pdf.

Abstract

In today's scenario, medical and dental professionals face a mammoth task while treating perplexing medical situations like organ failure or tissue loss. Though, different strategies exist to replace them, but ideal one is the same natural tissue or organ. In this aspect, stem cells have emerged in a promising way to provide an ideal replacement. There are different types of stem cells starting from the embryonic stage referred to as human embryonic stem cells to adult stem cells. Though in dentistry stem cell research is lagging as compared to the medical field but still a lot progress has been achieved in recent years. The stem cells have been isolated from dental pulp, human exfoliated deciduous teeth, and apical papilla and so on. These stem cells have provided exciting results like dentin-pulp regeneration, periodontal regeneration but ambiguity still prevails. As a result, much has to be further researched before its clinical application becomes a reality. Hence, these stem cells opened a new avenue in the field of regenerative dentistry.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3793402/.