Listed below are a few extracts which shed light on recent advances in medical research on dental pulp stem cell
Molecules Modulation of the Differentiation of Dental Pulp Stem Cells by Different Concentrations of β-Glycerophosphate
Department of Prosthodontics, the 2nd Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, China
Dentinogenesis is a necessary prerequisite for dental tissue engineering. One of the steps for dentinogenesis is to obtain large quantities of highly purified odontoblasts. Therefore, we have undertaken an experiment applying different concentrations of β-glycerophosphate (β-GP) to induce the differentiation of dental pulp stem cells (DPSCs) in a long-term 28-day culture. In the meanwhile, we have studied the time- and maturation-dependent expression of matrix extracellular phosphoglycoprotein (MEPE) and that of the odontoblast-like marker-dentin sialoprotein (DSP), in order to investigate an optimized mineralized condition. Western blot results revealed that the expression of DSP became lower when accompanied by the increase of the β-GP concentration, and there was also an influence on MEPE expression when different concentrations of β-GP were applied. Meanwhile, the mineralized groups had an inhibitory function on the expression of MEPE as compared with the control group. Above all, all experimental groups successfully generated mineralized nodules by Alizarin Red S and the 5 mM β-GP group formed more mineralized nodules quantitated using the CPC extraction method. In conclusion, there is a significant modulation of the β-GP during the differentiation of the DPSCs. The degree of odontoblast differentiation is β-glycerophosphate concentration dependent. A low concentration of β-GP (5 mM) has been shown to be the optimal concentration for stimulating the maturation of the DPSCs. Moreover, MEPE accompanied with DSP clearly demonstrates the degree of the differentiation.
Dlx3 and Dlx5 Inhibit Adipogenic Differentiation of Human Dental Pulp Stem Cells.
Dlx3 and Dlx5 are homeobox domain proteins and are well-known regulators of osteoblastic differentiation. Since possible reciprocal relationships between osteogenic and adipogenic differentiation in mesenchymal stem cells exist, we examined the regulatory role of Dlx3 and Dlx5 on adipogenic differentiation using human dental pulp stem cells. Over-expression of Dlx3 and Dlx5 stimulated osteogenic differentiation but inhibited adipogenic differentiation of human dental pulp stem cells. Dlx3 and Dlx5 suppressed the expression of adipogenic marker genes such as C/EBPalpha, PPARgamma, aP2 and lipoprotein lipase. Adipogenic stimuli suppressed the mRNA levels of Dlx3 and Dlx5, whereas osteogenic stimuli enhanced the expression of Dlx3 and Dlx5 in 3T3-L1 preadipocytes. These results suggest that Dlx3 and Dlx5 exert a stimulatory effect on osteogenic differentiation of stem cells through the inhibition of adipogenic differentiation as well as direct stimulation.
Coculture of Dental Pulp Stem Cells with Endothelial Cells Enhances Osteo-/Odontogenic and Angiogenic Potential in Vitro.
INTRODUCTION: Dental pulp stem cells (DPSCs) have received much attention as a promising population of stem cells in regenerative endodontics. Securing a good blood supply during regeneration is a challenging task because of the constricted apical canal opening, which allows only a limited blood supply. The aim of this study was to investigate any potential synergistic effects of dental pulp stem cells and endothelial cells (ECs) on osteo-/odontogenic and angiogenic differentiation in vitro.
METHODS:Different ratios of DPSCs and ECs were cultured in direct contact using optimized medium for coculture. The 70% confluent cocultures were incubated in the osteo-/odontogenic differentiation medium for up to 3 weeks. Alkaline phosphatase (ALP) activity, the expression levels of ALP, bone sialoprotein (BSP), dentin sialophosphoprotein (DSPP) genes, and alizarin red staining for mineralization at different time points were analyzed. The tubular network formation on Matrigel and the gene expression levels of CD117, VEGF, CD34, and Flk-1 were used as assays to analyze angiogenesis.
RESULTS:The quantification of ALP in DPSC:EC cocultures revealed a greater ALP activity compared with DPSC-alone cultures. At all the time points, 1:1 cultures showed a significantly greater ALP activity than that of DPSC-alone cultures. Alizarin red staining and quantification revealed a much greater amount of calcification in the 1:1 and 1:5 cocultures compared with other cultures (P < .01). The expression levels of ALP, BSP, and DSPP genes further confirmed the greater osteo-/odontogenic differentiation in cocultures compared with those of DPSC-alone cultures. Matrigel assay showed that the addition of DPSCs stabilized preexisting vessel-like structures formed by ECs and increased the longevity of them.
Conclusion: Direct coculture of DPSCs and ECs enhances the in vitro differentiation toward osteo-/odontogenic and angiogenic phenotypes
Mesenchymal Dental Stem Cells in Regenerative Dentistry
Cell Therapy Unit. Hospital Universitary Virgen de la Arrixaca, Faculty of Medicine and Odontology, University of Murcia, Spain Institute of Neurosciences. University Miguel Hernández. Alicante. Spain
In the last decade, tissue engineering is a field that has been suffering an enormous expansion in the regenera¬tive medicine and dentistry. The use of cells as mesenchymal dental stem cells of easy access for dentist and oral surgeon, immunosuppressive properties, high proliferation and capacity to differentiate into odontoblasts, cemen¬toblasts, osteoblasts and other cells implicated in the teeth, suppose a good perspective of future in the clinical dentistry. However, is necessary advance in the known of growth factors and signalling molecules implicated in tooth development and regeneration of different structures of teeth. Furthermore, these cells need a fabulous scaffold that facility their integration, differentiation, matrix synthesis and promote multiple specific interactions between cells.
In this review, we give a brief description of tooth development and anatomy, definition and classification of stem cells, with special attention of mesenchymal stem cells, commonly used in the cellular therapy for their trasdif¬ferentiation ability, non ethical problems and acceptable results in preliminary clinical trials.
In terms of tissue engineering, we provide an overview of different types of mesenchymal stem cells that have been isolated from teeth, including dental pulp stem cells (DPSCs), stem cells from human exfoliated deciduous teeth (SHEDs), periodontal ligament stem cells (PDLSCs), dental follicle progenitor stem cells (DFPCs), and stem cells from apical papilla (SCAPs), growth factors implicated in regeneration teeth and types of scaffolds for dental tissue regeneration.
Comparison of Gingiva, Dental Pulp, and Periodontal Ligament Cells From the Standpoint of Mesenchymal Stem Cell Properties
Orthopedic Surgery, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University, Tokyo, Japan
The specific properties of mesenchymal stem cells (MSCs) in oral tissues still remain unknown though their existence has been previously reported. We collected gingiva, dental pulp, and periodontal ligament tissues from removed teeth and isolated MSCs. These MSCs were compared in terms of their yields per tooth, surface epitopes, and differentiation potentials by patient-matched analysis. For in vivo calcification analysis, rat gingival and dental pulp cells mounted on β-tricalcium phospateTCP were transplanted into the perivertebral muscle of rats for 6 weeks. Gingival cells and dental pulp cells showed higher yield per tooth than periodontal ligament cells (n=6, p < 0.05). Yields of periodontal ligament cells were too low for further analysis. Gingival and dental pulp cells expressed MSC markers such as CD44, CD90, and CD166. Gingival and dental pulp cells obtained phenotypes of chondrocytes and adipocytes in vitro. Approximately 60% of the colonies of gingival cells and 40% of the colonies of dental pulp cells were positively stained with alizarin red in vitro, and both gingival and dental pulp cells were calcified in vivo. We clarified properties of MSCs derived from removed teeth. We could obtain a high yield of MSCs with osteogenic potential from gingiva and dental pulp. These results indicate that gingiva and dental pulp are putative cell sources for hard tissue regeneration.
Characterisation of Dental Pulp Stem Cells: A New Horizon for Tissue Regeneration?
Pulp Biology and Endodontics, Oral Restitution Department, Graduate School of Medical and Dental Sciences, Tokyo.
Stem cells possess multipotent properties that allow them to differentiate into various cells, which may be potentially useful in tissue regeneration. Stem cell populations are reported to be present in various tissues of hematopoietic, neural and mesenchymal lineages, with the presence of stem cell populations in dental pulp tissue first described more than 10 years ago. The main components of dental pulp tissue are dental pulp cells, which are mesenchymal cells derived from the neural crest.(1,2) Some of these cells demonstrate high growth potential and possess multiple differentiation properties, and have been designated dental pulp stem cells (DPSCs). These cell populations are present not only in adult pulp tissue, but also in deciduous tooth pulp and apical papilla. DPSCs isolated by different methods, such as high growth potential, using various surface markers, and high efflux of a fluorescent nuclear stain (Hoechst 33342), all show multipotency, however their surface marker expression is somewhat different from each other. In vivo studies have revealed the possibility use of DPSCs in the regeneration of various tissue. DPSCs are of dental pulp origin, and dental pulp tissue is regenerated from DPSCs. Many researchers have focused on the dentine- and bone-forming properties of DPSCs, but their neuronal and muscular differentiation capacity suggests they may have a wider clinical application.
Mineral Trioxide Aggregate Promotes Odontoblastic Differentiation via Mitogen-Activated Protein Kinase Pathway in Human Dental Pulp Stem Cells.
Mitogen-activated protein kinase (MAPK) pathways are involved in stem cell differentiation. However, the odontoblastic differentiation-inducing effects by mineral trioxide aggregate (MTA) via MAPK pathways have not been clarified in human dental pulp stem cells (DPSCs). In this study we investigated the effects of MTA on cell viability and production of differentiation markers, and the involvement of MAPK signaling pathways in cultured human DPSCs. Cells were cultured with MTA, and the viability and differentiation productions of the cells were determined using the MTT assay and real-time PCR analysis, respectively. MAPK activation was measured by western blotting. MTA at concentrations of 20 and 10 mg/ml was toxic for human DPSCs. MTA significantly increased the expression of alkaline phosphatase (ALP), dentin sialophosphoprotein (DSPP), type I collagen (COLI), osteocalcin (OCN) and bone sialoprotein (BSP) mRNAs and induced the phosphorylation of p42 and p44 (p42/44), p38 and c-Jun N-terminal kinases 1 and 2 (JNK1/2) MAPK. Furthermore, the inhibitor of p42/44 MAPK attenuated the MTA-induced odontoblastic differentiation. These data indicated that MTA-induced odontoblastic differentiation of human DPSCs was via MAPK pathways, which may play a key role in the repair responses of dentin-pulp-like complexes.
High-Purity Hepatic Lineage Differentiated from Dental Pulp Stem Cells in Serum-Free Medium.
Department of Oral Health, Nippon Dental University, Tokyo, Japan.
Mesenchymal cells from human deciduous and extracted third molar pulp were isolated and expanded in vitro. We separated CD117-positive cells by using a magnetic-activated cell sorter. The cells were characterized immunofluorescently by using known stem cell markers. For hepatic differentiation, the media were supplemented with hepatic growth factor, insulin-transferrin-selenium-x, dexamethasone, and oncostatin M. Expression of hepatic markers alpha fetoprotein, albumin, hepatic nuclear factor-4 alpha, insulin-like growth factor-1, and carbamoyl phosphate synthetase was examined immunofluorescently after differentiation. The amount of differentiated cells was assessed by using flow cytometry. Glycogen storage and urea concentration in the medium were defined.
RESULTS:Both cell cultures demonstrated a number of cells positive for all tested hepatic markers after differentiation, ie, albumin-positive cells were almost 90% of differentiated deciduous pulp cells. The concentration of urea in the media increased significantly after differentiation. Significant amount of cytoplasmic glycogen storage was found in the cells.
CONCLUSIONS: Without serum both cell types differentiated into high-purity hepatocyte-like cells. These cells offer a source for hepatocyte lineage differentiation for transplantation in the future.
Odontogenic Differentiation of Human Dental Pulp Stem Cells Stimulated by the Calcium Phosphate Porous Granules
Sunyoung Nam, Jong-Eun Won, Cheol-Hwan Kim, and Hae-Won-Kim
Effects of three-dimensional (3D) calcium phosphate (CaP) porous granuales on the growth and odontogenic differentiation of human dental pulp stem cells (hDPSCs) were examined for dental tissue engineering. hDPSCs isolated from adult human dental pulps were cultured for 3-4 passages, and populated on porous granules. Cell growth on the culture dish showed an ongoing increase for up to 21 days, whereas the growth on the 3D granules decreased after 14 days. This reduction in poliferative potential on the 3d granules was more conspicuous under the osteogenic medium conditions, indicating that the 3D granules was confirmed by the increased alkaline phosphatase activity, up-regulation of odontoblast-specific genes, including dentin sialophosphoprotein (DSPP) and dentin matrix protein 1 (DMP1) by quantitative polymerase chain reaction, and greater level if dentin sialoprotein synthesis by western blot. Moreover, the cellular mineralization, as assessed by Alizarin red S and calcium quantification, was significantly higher in the 3D CaP granules than in the culture dish. Taken all, the 3D substrate conditions for cell growth and odontogenic development.
Human Dental Pulp Stem Hook into Biocoral Scaffold Forming an Engineered Biocomplex
Carlo Mangano, Francesca Piano, Riccardo d' Aquino, Alfredo De Rosa, Giovanna lezzi, Adriano Piatelli, Luigi Liano, Thimios Mitsiadis, Vincenzo Desiderio, Francesco Mangano, Gianpaolo Papaccio, Virginia Tirino
Tha aim of this study was to elevate the behavior of human Dental Pulp Stem Cells (DPSCs), as well as human osteoblast, when challenged on a Biocoral Scaffold, which is a porous natural hydroxyapatite. For this purpose human DPSCs were seeded onto a three- dimensional (3D) Biocoral Scaffold or on flask surface(control). Either normal rotative (3D) cultures were performed. Scanning electron microscopic analyses, at 8, 24 and 48 h of the culture showed that cells did not adhere on the external surface, but moved into the cavities inside the Biocoral structure. After 7, 15, and 30 days of culture, morphological and molecular analyses suggested that the Biocoral Scaffold leads DPSCs to hook into the cavities where these cells quickly start to secrete the extra cellular matrix(ECM) and differentiate into osteoblasts. Control human osteoblasts also moved into internal cavities where they secreted the ECM. Histological section revealed a diffused bone formation inside the Biocoral samples seeded with DPSCs or human osteoblasts, where the orginal scaffold and the new secreted biomaterial were completely integrated and cell were found within the remaining cavities. In addition, RT-PCR analyses showed a significant increase of osteoblast- related gene expression and, above all, of those genes highly expressed in mineralized tissues, includinh osteocalcin, OPN and BSP. Furthermore, the effects on the interaction between osteogenesis and angiogenesis were observed and substanciated by ELISA assays. Taken together, our results provide clear evidence that DPSCs differentiated into osteoblasts, forming a biocomplex made of Biocoral, ECm and differentiated cells.
Induced Invitro Differentiation of Neural – like Cells from Human Exfoliated Deciduous Teeth-Derived Stem Cells
Pediatric Department, School of Dentistry, Department of Anatomical Sciences, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran
Stem cells from human exfoliated deciduous teeth (SHED) are highly proliferative, clonogenic and multipotent stem cells with a neural crest cell orgin. Additionally, they can be collected with minimal invasiveness in comparison with other sources of mesenchymal stem cells (MSCs). Therefore ,SHED could be a desirable option for potential therapeutic application. In this study, SHEDs were established from enzyme-disaggregated deciduous dental pulp obtained from 6 to 9 year- old children. The cell has typical fibroblastoid morphology and expressed antigens characteristic of MSCs, STRO1, CD146, CD45, CD90, CD106 and Cd166, but not the hematopoietic and endothelial markers, CD34 and CD31, as assassed by FACS analysis. Differentiation assessment revealed a strong osteogenic and adipogenic potential of SHEDs. In order to further evaluate the in vitro differentiation potential of SHED into neural cells, a simple short time growth factor-mediated induction was used. Immunoflurescence staining and flow cytometric analysis revealed that SHED rapidly expressed nestin and β-III tubulin, and later expressed intermediate neural markers. In addition, the intensity and percentages of nestin and β-III tubulin and mature neural markers(PSA-NCAM,NeuN, Tau,TH or GFAP) increased significantly following the treatment. Moreover, RT-PCR and Western blot analyses showed that the neural markers were strongly unregulated after induction. In conclusion, these results provide evidence that SHED can differentiate into neural cells by the expression of a comprehensive set of genes and proteins that define neural-like cells in vitro. SHED cells might be considered as new candidates for the autologous transplantation of a wide variety of neurological diseases and neurotraumatic injuries.
Isolation of Pluripotent Stem Cells from Human Third Molar Dental Pulp
M. Atari, M. Barajas, F. Hernandez-Alfaro, C. Gil, M. Fabregat, E. Ferres Padro, L. Giner, L. Casals
Regenerative Medicine Laboratory, Universitat Internacional de Catalunya, Barcelona, Spain.
Potent stem/progenitor cells have been isolated from normal human dental pulps, termed dental pulp stem cells (DPSCs). However, no study has described the presence of stem cell populations in human dental pulp fron the third molar with embryonic phenotypes. The dental pulp tissue was cultured in media with the presence of LIF, EGF, and PDGF. In the present study, we describe a new population of pluripotent stem that were isolated from dental pulp (DPPSC). These cells are SSEA-4+ , Oct4+, Nanog+, FLK-1+, HNF3beta+, Nestin+, Sox2+, Lin28+, c-Myc+, CD13+, CD105+, CD34-, CD45-, CD90low, CD29+, CD73low, STRO-1low and CD146-. We have investigated by SEM analysis and q-RT-PCR the capacity of DPPSCs to 3D differentiate in vitro using the CELL Carrier 3D glass scaffold into tissues that have similar characteristics to embryonic mesoderm and endoderm layers. These data would support the use of these cells, which are derived from an easily accessible source and can be used in future regeneration protocols formany tissue types that differentiate from the three embryonic layers.
Stem Cell Research and Theraphy
Immunomodulatory properties of stem cells from human exfoliated deciduous teeth
Takayoshi Yamaza , Akiyama Kentaro , Childer chen , Yi Liu , Yufang Shi , Stan Gronthos , Songlin Wang ,Songato Shi
Introduction : Stem cells from human exfoliated deciduous teeth (SHED) have been identified as a population of postnatal stem cells capable of differentiating into osteogenic and odontogenic cells , adipogenic cells, and neural cells. Herein we have characterized mesenchymal stem cell properties of SHED in comparison to human bonel marrow mesenchymal stem cells(BMMSCs).
Methods: We used in vitro stem cells analysis approaches, including flow cytometry, inductive differentiation, , telomerase activity, and Western Blot analysis multipotent differentiation of SHED and in vivo implantation to assess tissue regeneration of SHED. In addition, we utilized systemic SHED transplantation to treat systemic lupus erythematosus (SLE)-like MRL/lpr mice.
Results: We found that SHED are capable of differentiating into osteogenic and adipogenic cells, expressing mesenchymal surface molecules (STRO-1, CD146, SSEA4, CD73, CD105, and CD166), and activating multiple signaling pathways, including TGFβ, ERK, Akt, Wnt, and PDGF. Recently, BMMSCs were shown to possess an immunomodulatory function that leads to successful therapies for immune diseases. We examined the immunomodulatory properties of SHED in comparison to BMMSCs and found that SHED transplantation is capable of effectively reversing SLE-associated disordes in MRL/lpr mice. At the cellular level, SHED transplantation elevated the ratio of regulatory T cells (Tregs) via Th 17 cells.
Conclusion: These data suggest that SHED are an accessible and feasible mesenchymal stem cell source for treating immune disordes like SLE.
Expression of Multiple Stem Cell Markers in Dental Pulp Cells Cultured in Serum-free Media
Viability tests showed a significant difference between the control and SFMs in the deciduous tooth pulp cells(DTPCs) and wisdom tooth pulp cells (WTPCs). However all SFMs demonstrated 84%-90% viability, whereas the control showed 90%-93% . In both DTPCs and WTPCs, SFM#1 had the highest proliferation rate among the 4 SFMs. Immunocytochemistry stained positive stem cell markers most intensely in cells cultured with SFM#1. Furthermore , all stem cell markers for ectoderm, mesoderm and endoderm were expressed in the cell cultured with SFM#1.
Differentiation Potential of STRO- 1+ Dental Pulp Stem Cell Changes During Cell Passaging
Background: Dental pulp stem cells(DPSCs) can be driven into odontoblast, osteoblast and chondroncyte lineages in different inductive media. However, the differentiation potential of native DTPs after serial passaging in the routine culture system has not been fully elucidated.
Result: DPSCs were isolated from human/ rat dental pulp by the magnetic activated cell sorting based on STRO-! Expression, cultured and passaged in the conventional culture media. The biological features of STRO-1+ DPSCs at the 1st and 9th passages were investigated. During the long-term passage, the proliferation ability of human STRO-1+ DPSCs was down regulated as indicated by the growth kinetics. When compared with STRO-1+ DPSCs at the 1st passage(DPSC-P1), the expression of mature osteoblast-specific genes /proteins(alkaline phosphatase, bone sialoprotein, osterix and ostiopontin), odontoblast-specific gene/protein (dentin sialophosphoprotein and dentin sialoprotein), and chondrocyte specific gene/protein(type II colagen) was significantly upregulated in human STRO-1+ DPSCs at the 9 th passage (DPSC-P9). Further more human DPSC-P9 cells in the mnineralization-inducing media presented higher level of alkaline phosphate at day3 and day 7 respectively, and produced more mineralized matrix than DPSC-P9 cells at day 14. In Volvo transplantation results showed that rat DPSC-P1 cell pellets developed into dentin, bone and cartilage structure respectively, while DPSC-P9 can only generate bone tissues.
Conclusion: These findings suggest that STRO-1+ DPSCs consists of several interrelated subpopulations which can spontaneously differentiate into odondoblast, osteoblast and chondrocytes. The differentiation capacity of these DPSCs changes during cell passaging, and DPSCs at the 9th passage restrict their differentiation potential to the osteoblast lineage in vivo.
Isolation and In Vitro Characterization of Dental Pulp Stem Cell from Natal Teeth
Dental pulp stem cells were primarily derived from the pulp tissue of exfoliated deciduous teeth, primary incisor and permanent third molar teeth. The aim of this study was to isolate and extensively characterize SCs derived from human natal dental pulp (hNDP). For characterization , proliferation capacity, phenotypic properties, ultrastructural and differentiation characteristics and gene expression profiles were utilized. A comparison was done between the properties of NDP-SCs and the properties of mesenchymal stem cells (MSCs) from bone marrow (BM) of the human. Stem cells isolated from hNDP and hBM were analysed by flow cytometry, reverse transcriptase- PCR, Real Time-PCR, and immunocytochemistry. Both cell lines were directionally differentiated towards adipogenic, osteogenic, chondrogenic, myogenic and neurogenic lineages. hNDP-SCs and hBM-MSCs expressed CD13, CD44, CD90, CD146 and CD166, but not CD3, CD8, CD11b, CD14, CD15, CD19, CD34, CD45, CD117, and HLA-DR. Ultrastructural characteristics of hNDP-SCs showed more developed and metabolically active cells. hNDP-SCs and hBM-MNSCs expressed some adipogenic(leptin, adipophilin and PPARγ), myogenic (desmin, myogenin,myosinlla, and α-SMA), neurogenic (γ-enolase,MAP2a,b, c-fos, nestin, NF-H, NF-L, GFAP and betalll tubulin), osteogenic (osteonectin, osteocalcin, osteopontin, Runx-2 and type I collagen) and chondrogenic (typeII collagen, SOX9) markers without any stimulation towards differentiation under basal conditions. Embryonic stem cell markers Oct4, Rex-1, FoxD-3, Sox2, and Nanog were also identified. The differentiation potential of hNDP-SCs and hBM-MSCs to adipogenic, osteogenic, chondrogenic, myogenic and neurogenic was shown. This report described the first successful isolation and characterization of hNDP-Scs.
Dental Pulp Stem Cells and their Characterization
Jakub Suchanek, Thomas Soukub, Benjamin Visek, Romana Ivancakova, Lenka Kucerova, Jaroslav Mokry
Aims: Our aims were to isolate dental pulp stem cells, to cultivate them in various media and to investigate their basic biological properties and phenotype.
Methods: 16 lines of dental pulp stem cells (DPSCs) were isolated form an impacted third molar. After enzymatic dissociation of dental pulp, DPSCs were cultivated in modified cultivation media for mesenchymal adult progenitor cells containing 2 % or 10 % fetal calf serum (FCS), or in modified 2 % FCS cultivation media supplemented with ITS. Cell viability and other biological properties were examined periodically using a Vi-Cell analyzer and Z2-Counter. DNA analysis and phenotyping were done using flow cytometry
Results: We were able to cultivate DPSCs in all tested cultivation media over 40 population doubling. Our results showed that DPSCs cultivated in medium supplemented with ITS had shorter average population doubling time (24.5,15.55-35.12 hours) than DPSCs cultivated in 2 % FCS (55.43,21.57-187.14 hours) or 10 % FCS (42.56, 11.86-101.3 hours). Cell diameter was not affected and varied from 15 to 16 µm. DPSCs viability in the 9th passage was over 90 % Our phenotypical analysis was highly positivity for CD29, CD44, CD90 and HLA I, and negative for CD34, CD45, CD71, HLA ll. DPSC lines cultivated in all media showed no signs of degeneration or spontaneous differentiation during the expansion process.
Conclusion: We showed that ITS supplement in the cultivation media greatly increased the proliferative activity of DPSCs. Other DPSCs biological properties and phenotype were not affected.
Human Immature Dental Pulp Stem Cells Share Key Characteristic Features with Limbal Stem Cells.
Monteiro Bg, Serafim RC, Melo Gb, Silva MC, Lizier NF, Maranduba CM, Smith RL, Kerkis A, Cerruti H, Gomes Ja, Kerkis I.
We have demonstrated, using immunohistochemistry and transcription-polymerase chain reaction LSC, such as ABCG2,integrin beta1, vimentin, p63,connexin 43 and cytokeratins 3/12 were also capable of reconstructing the eye surface after induction of unilateral TLSCD in rabbits, as shown by morphological and corneal epithelium. Our data suggest characteristics with Lsc and might be used as a potential alternative source of cells of corneal reconstruction
Therapeutic Window of Stem Cell Potential for Targeting Pediatric Malignant Brain Tumors: An Opportunity for Stem Cell Therapy
Shengwen Calvin Li . Yuan-Ping Han
Brent A. Dethlefs . William Gunter Loudon
Stem cells, with their natural ability to seek out brain tumors, could be used to accurately deliver therapy directly to the cancer sparing normal tissues for suppression of tumor growth.
Evaluation of Pluripotency in Human Dental Pulp Cells
Purpose: Postnatal stem cells have been isolated from various tissues, including bone marrow, neural tissue, skin, reina, and dental epithelium. Recently, adult stem cells have been isolated from human dental pulp. Postnatal stem cells have been isolated from a variety of tissues. Previously, it was generally accepted that the differentiation potential of postnatal stem cells was Lineage restricted.
Materials and Methods: Normal impacted third molars were collected from adults and normal exfoliated deciduous teeth(SHED; stem cells from human exfoliated deciduous teeth) by single-colony selection and magnetic activated cell sorting.
Results: BMP-2 treatment groups produced alkaline phosphatase in the cells and also produced and secreted osteocalcin in the culture medium, and were capable of including an upregulated expression of Osteocalcin or Sox9, Col 2, and Col X by reverse transcriptase polymerase chain reaction(RT-PCR). For adipogenic differentiation, there is potential for SHED and dental pulp stem cells(DPSC) to express to adipocyte-specific transcripts, PPRAγ2 and LPL, in vitro, as do bone marrow mesenchymal stem cells by RT-PCR.
Conclusion: This study demonstrated that pluripotential cells isolated from the pulp of human teeth expanded in vitro and differentiated into osteoblasts, chondrocytes and adipocytes. DPSC and SHED are not only derivated from a very accessible tissue resource but also capable of providing enough cells for potential clinical applications.
Articles on Therapeutic Applications
Therapeutics of stem cells in periodontal regeneration
Journal of Natural Science , Biology and Medicine , Jan 2011, Vol 2, Issue 1
Rajiv Saini et al
The structure and composition of the periodontium are affected in many acquired and hentable diseases and the most significant among these is periodontal diseases .periodontal regeneration is considered to be organically promising but clinically capricious. The principal requirements for tissue engineering are incorporation of appropriate numbers of responsive progenitor cells and the presence of bioactive levels of regulatory signals within an appropriate extracellular matrix or carrier construct. Stem tissues that are damaged And recent progress in stem cells research and in tissue engineering promises nowel prospects for therapeutic strategies for the replacement of a diseased or damaged tooth.
Intravenous Mesenchymal Stem Cells Prevented Rejection of Allogeneic Corneal Transplants by Aborting the Early Inflammatory Response
Molecular Therapy, (28 August 2012)|doi:10.1038/mt.2012.165
Joo Youn Oh, Ryang Hwa Lee, Ah Young Ko, Gavin W Roddy and Darwin J Prockop
Mesenchymal stem/progenitor cells (MSCs) were reported to enhance the survival of cellular and organ transplants. However, their mode of action was not established. We here used a mouse model of corneal allotransplantation and demonstrated that peri-transplant intravenous (i.v.) infusion of human MSCs (hMSCs) decreased the early surgically induced inflammation and reduced the activation of antigenpresenting cells (APCs) in the cornea and draining lymph nodes (DLNs). Subsequently, immune rejection was decreased , and allograft survival was prolonged. Quantitative assays for human GAPDH revealed that 10 hMSCs out of 1 x 106 injected cells were recovered in the cornea 10 hours to 28 days after i.v. infusion. Most of hMSCs were trapped in lungs where they were activated to increase expression of the gene for a multifunctional anti-inflammatory protein tumor necrosis factor-α stimulated gene/protein 6 (TSG-6). I.V. hMSCs with a knockdown of TSG-6 did not suppress the early inflammation and failed to prolong the allograft survival. Also, i.v. infusion of recombinant TSG-6 reproduced the effects of hMSCs. Results suggest that hMSCs improve the survival of corneal allografts without engraftment and primarily by secreting TSG-6 that acts by aborting early inflammatory responses. The same mechanism may explain previous reports that MSCs decrease rejection of other organ transplants.
High-purity hepatic differentiated from dental pulp stem cells in serum-free medium
J Endod 2012 Apr;38(4):475-80. Epub 2012 Jan28
INTRODUCTION: We have previously differentiated hepatocyte like cells from deciduoud tooth pulp stem and extracted third molar pulp stem cells with a protocol that used fetal bovine serum, but it showed high contaminations of nondifferentiated cells. Both the lower purity of hepatically differentiated cells and usage of serum are obstacles for applications of cell therapy or regenerative medicine. Objective of this study was to investigate the capacity for and purity of hepatocyte-like differentiation of CD 117-positive dental pulp stem cells without serum.
METHODS: Mesenchymal cells from human deciduous and extracted third molar pulp were isolated and expanded in vitro. We separated D117-position cells by using a magnetic-activated cell sorter. The cells were characterized immunofluorescently by using known stem cell markers. For hepatic differentiation, the media were supplemented with hepatic growth factor, insulin transferring-selenium-x, dexamethasone, and oncostatin M. Expression of hepatic markers alpha fetoprotein, albumin, hepatic nuclear factor-4 alpha, insulin-like growth factor-1, and carbamoyl phosphate synthetase was examined immunoflurescently after differentiation. The amount of differentiated cells was assessed by using flow cytometry. Glycogen storage and area concentration in the medium were defined.
RESULTS: Both cell cultures demonstrated a number of cells positive for all tested hepatic markers after differentiation,ie, albumin-positive cells were almost 90% of differentiated deciduous pulp cells. The concentration of urea in the media increased significantly after differentiation. Significant amount of cytoplasmic glycogen storage was found in the cells.
CONCLUSIONS: Without serum both cell types differentiated into high-purity hepatocyte-like cells. These cells offer a source for hepatocypte lineage differentiation for transplantation in the future.
Stem Cells in Regenerative Endodontics: A Review
Regenerative Endodontics is expected to become the next big treatment revolution. It is the creation and delivery of tissues to replace a diseased, missing and traumatized pulp dentin complex, in addition to tooth supporting tissues. It employs dental stem cells for this which is especially important because they are a type of mesenchymal stem cells that produce more neurotrophic factors than other cell types. These cells have demonstrated the ability to protect and repair neural tissue in diseases like spinal cord injury and stroke and hence can also be used for development of treatment solutions for such diseases. Dental stem cells are also easy to procure and preserve for use in future therapies.
Prespectives on Mesenchymal Stem Cell: Tissue Repair, Immune Modulation, and Tumor Homing
Arch Pharm Res Vo 35, No,2 201-211, 2012
DOT 10.1007 / s 12272-012-0201-0
Hyun Sook Hong, Yeong Hoon Kim, Youngsook Son
Mesenchymal stem cells (MSCs) or MSC – Like cells have been identified in a variety of different tissues that share molecular expression profiles and biological functions but also retain a unique differentiation preference depending on their tissue origins. MSCs play beneficial roles in the healing of damaged tissue by directly differentiating to many different resident cell types and / or by secreting several trophic factors that aid tissue repair. Aside from MSCs reparative stem cell function, they drive immune responses toward immunosuppression and anti - inflammation . This novel function of MSCs opens up new avenues for clinical development of MSC immune - therapeutics to treat uncontrollable, life threatening, severe , chronic inflammation and autoimmune disease.
MSCs or MSC – like cells, even though their tissue origins are different, are quite similar in expression of molecular markers and biological functions. MSCs are expected as reparative stem cell for a variety of tissue injuries or disease by differentiating to many different types of resident tissue cells. More frequently, life threatening steroid refractory immune rejection, and autoimmune syndrome, MSCs other novel function, an immune modulating effect, is adopted for the therapeutic rationale. However, in certain tumor setting, MSCs immune modulatory function, in combination with their tumor homing preference, may provide a reason for their causing unwanted tumor progression. This tumor homing capacity of MSCs can be strategically utilized as a novel tumor – targeting anticancer therapeutic. In conclusion, recent progress in MSCs therapeutics promises benefits for numerous uncontrolled diseases that cannot be met by conventional medication. Also their possible risk for the progression of tumors was uncovered. Therefore, MSC therapeutics can be advantageously developed as a specific tumor – targeting delivery vehicle.
ANGIOGENIC POTENTIAL OF HUMAN DENTAL PULP STEM CELLS FOR SKELETAL TISSUE ENGINEERING
Journal of Bone & Joint Surgery, British Volume
J Bone Joint Surg Br 2012 vol, 94-B no. SUPP XXXVI 43
NAL-Hazaimeh, J Beattle, M Duggal and XB Yang
Angiogenesis and the ability to provide appropriate vascular supply are crucial for skeletal tissue engineering. The aim of this study was to investigate the anglogenic potential of human dental pulp stromal cells (HDPSCs) and stro-1 positive populations as well as their role in tissue regeneration ( the clinical reality).
HDPSC were isolated from the pulp tissue of human permanent teeth by collagenase digestion. STRO-1 positive cells were enriched using mono clonal anti STRO-1 and anti CD-45 PE conjugated anti bodies together with and fluorescence activated cell sorting (FACS). Cells isolated by FACS were grown to passage 4 and cultured as mono layers or on 3D matrigel scaffold in endothelial cell growth medium -2 (EGM_2) with/without 50ng/ml of vascular endothelial growth factor (VEGF). Cells cultured in alpha MEM supplemented with 10% FCS were used as controls. After 24, 48 and 72 hours anglogenic marker expression (CD31, CD34, vWF and VEGFR-2) was determined by qRT-PCR and immune-histo chemistry.
Using three different donors 0.5-1.5% of total HDPSCs population was characterized as STRO_1+/CD45- cells at each time point cells cultured as mono layer in EGM-2 with VEGF showed up regulation of CD31 and VEGFR-2 expression compared to the control group while expression of CD34 and vWF remained unaffected . however on matrigel, all four genes were up regulated to different extents. CD31 and VEGFR-2 were up regulated to a greater degree compared to CD34 and vWF. Changes in gene expression in both cell types were time dependent. Immune-histochemical staining confirmed that the HDPSCs cultured in th test group showed positive staining for the four angiogenic markers ( CD31, CD34, vWF and VEGFR-2) when grown in both mono layer and 3D matrigel culture compared to control cultures. When cultured on matrigel ( but not monolayer) for 7 days, HDPSC formed tube like structures in the VEGF treated group.
This indicates the potential of use HDPSCs and their STRO-1 positive population for angiogenesis to enhance skeletal tissue repair and/or regeneration toward translational research for clinical benefit.
Mesenchyamal dental stem cells in regenerative dentistry
Journal section: Biomaterials and Bioengineering in Dentistry
Doi: 10. 4317 medoral 17925
Mesenchymal Dental Stem cells derived from teeth are easily accessible multipotent cells with the capacity to differentiate into distinct cell types. This new source of stem cells could be of benefit in cellular therapy and the eventual development of techniques for use in regenerative dentistry and degenerative diseases.
Role of mesenchymal stem cell therapy in Crohn's disease
Jignesh Dalal, Kimberly Gandy & Jos Domen
Pediatric Research (2012) 71, 445-451 doi: 10.1038/pr.2011.56
08 February 2012
Many trials of mesenchymal stem cells (MSCs) have been published in the past 5-6 y. MSCs inhibit T-cell alloreactivity in vitro by soluble factors and direct cell-to-cell contact. They are safe to infuse in humans with no acute toxicity and no ectopic tissue formation. Promising results of MSC infusion for graft-vs-host disease and fistulizing Crohn's disease (CD) have been published. Treatment of CD requires a comprehensive treatment approach to maintain Symptomatic control, improve health-related quality-of-life measures, and minimize complications from the disease. In this review, we will discuss the results of clinical trials using a novel treatment in the form of MSCs for treatment of CD and related complications. Success of these phase l, ll and lll trials have set the stage for usage of this novel treatment for children with CD.
The current and future Therapies of Bone Regeneration to Bone Repair Defects
International Journal of Dentistry
Volume 2012, Article ID 148261, 7 pages
Eijiro Jimi,1.2 Shizu Hirata , 3Kenji Osawa,1 Masamichi Terashita,4
Chiaki Kitamura,2.3 and Hidefumi Fukushima 1
Bone defects often result from tumour resection, congenital malformation, trauma, fractures, surgery, or periodontitis in dentistry. Although dental implants serve as an effective treatment to recover mouth function from teeth defects, many patients do not have the adequate bone volume to build an implant. The gold standard for the reconstruction of large bone defects in the use of autogeneous bone grafts. While autogeneous bone grafts is the most effective clinical method, surgical stress to the part of the bone being extracted and the quantity of extractable bone limit this method. Recently mesenchymal stem cell-based therapies have the potential to provide an effective treatment of osseous defects.
Although regenerative medicine has been tried in various fields, there is much demand for regenerative medicine in dentistry, particularly in bone regeneration. Depending on the state of periodontitis or jaw resection, it might take more than 6 to 12 months for occlusal reconstitution. Thus, the development of an efficient and high-quality bone derivation method is necessary. Cell-based therapy pave the way to rejection-free regenerative treatment for bone defects.
Stem cell therapy in oral and maxillofacial region: An Overview
Journal of Oral and Maxillofacial Pathology
Vol 16 issue 1 Jan – apr 2012 58
Sunil PM, Manikandan R 1, Muthu MS2, Abraham S3
The advantages of stem cells from oral and maxillofacial region is that
1. Have high plasticity. 2. It can be cryopreserved for longer period (Ideal for stem cell banking). 3. It showed good interaction with scaffold and growth factors. 4. Stem cells transplantations can cause pathogen transmission and also need immunosuppression, so autologous stem cell source is the best option. Dental pulp stem cells will be better fitting tool due to easy surgical access, the very low morbidity of the anatomical site after the collection of the pulp.
Human Dental Pulp-Derived Stem Cells Promote Locomotor Recovery after Complete Transection of the Rat Spinal Cord by Multiple Neuro-Regenerative Mechanisms
Kiyoshi Sakai, Akihito Yamamoto
Department of Oral and Maxillofacial Surgery, Department of Biochemistry, and Department of Orthopedic Surgery, Nagoya University Graduate School of Medicine, Nagoya, Japan.
Spinal cord injury (SCI) often leads to persistent functional deficits due to loss of neurons and glia and to limited axonal regeneration after injury. Here we report that transplantation of human dental pulp stem cells into the completely transected adult rat spinal cord resulted in marked recovery of hind limb locomotor functions. Transplantation of human bone marrow stromal cells or skin-derived fibroblasts led to substantially less recovery of locomotor function. The human dental pulp stem cells exhibited three major neuroregenerative activities. First, they inhibited the SCI-induced apoptosis of neurons, astrocytes, and oligodendrocytes, which improved the preservation of neuronal filaments and myelin sheaths. Second, they promoted the regeneration of transected axons by directly inhibiting multiple axon growth inhibitors, including chondroitin sulfate proteoglycan and myelin-associated glycoprotein, via paracrine mechanisms. Last, they replaced lost cells by differentiating into mature oligodendrocytes under the extreme conditions of SCI. Our data demonstrate that tooth-derived stem cells may provide therapeutic benefits for treating SCI through both cell-autonomous and paracrine neuroregenerative activities.
Clinical Outcomes of xeno-free autologous cultivated limbal epithelial transplantation :a 10-year study
Virender S Sangwan , Sayan Basu, Geeta K Vemuganti , Kunjal Sejpal , Sandhya V subramaniam , Souvik Bandyopadhyay , Sannapaneni Krishnaiah , Subhash Gaddipai, Shubha Tiwari, Dorairajan Balasubramanian
September 2, 2011 as 10.1136/bjophthalmol-2011-300352
LV Prasad Eye
Institute Efficacy of xeno-free autologuous cell- based treatment of limbal stem cell deficiency. A completely epithilised , avascular and clinically stable coreneal surface was seen in 142 of 200(71%) eyes at a mean follow-up of 3 + or – 1.6(range :1-7.6)years. Over the last two decades the surgical techniques has evolved from direct limbal transplantation to transplantation of ex vivo cultivated limbal epithelical cells, limbal cultivation may involve use of xenogenic tissue , associated with the risk of transmission of known or unknown infections to the transplant recipient of murine feeder cells, fetal bovine serum(FBS) or animal-derived growth factors. We report the long term survival, visual benefit and complications of transplantation of autologous limbal epithelial cells.
Stem cells in clinical practice: applications and warnings
Lodi et al. Journal of Experimental & Clinical Cancer Research 2011, 30:9
Daniele Lodi1 , Tommaso lannitti2*, Beniamno Palmieri3
A cell polarizes itself, so that cell-fate determinant molecules are specifically localized on one side. After that, the mitotic spindle aligns itself perpendicular to the axis polarity. End of the process two different cells are obtained [5-7]. Promising results have been described in disorders, such as diabetes  and neurodegenerative diseases [274, 275], where SCs graft can reestablish one or more deficit cellular lineages and, generally, a healthy state. Notably, many clinical studies have underlined the immunomodulatory effect of SCs in autoimmune diseases, such as multiple sclerosis , organ transplants  and in uncontrolled immune-inflammatory reactions [277-279].
Cardiovascular Applications of Stem Cell Therapy
Soheil Sadri1 , Ramesh Mazhari2, Maziar Sadri2, Nahal Konjedi1, and Palak Shah2*
Sadri et al.. J Stem Cell Res Ther 2011. S1
The prospect of cell-based therapies for cardiovascular disease has led to excitement within the scientific community as cardiovascular disease continues to be the leading cause of death worldwide. There is a growing body of evidence from both the basic sciences as well as through translational studies that prove the ability of stem cells to improve left ventricular function.
Mesenchymal stem cell-based therapies in regenerative medicine: application in rheumatology
Marie Maumus 1.2, David Guerit 1.2 , Karine Toupet 1.2, Christan Jorgensen 1.2.3 and Daniele Noel*1.2.4
Maumus et al. Stem Cell Research & Therapy 2011, 2:14 http://stemcellres.com/content/2/2/14
Growing knowledge on the biology of mesenchymal stem cells (MSCs) has provided new insights into their potential clinical applications, particularly for rheumatologic disorders. Historically, their potential to differentiate into cells of the bone and cartilage lineages has led to a variety of experimental strategies to investigate whether MSCs also display immunosuppressive properties, which have prompted research on their capacity to suppress local inflammation and tissue damage in a variety of inflammatory autoimmune diseases and, in particular, in rheumatoid arthritis. Currently, an emerging field of research comes from the possibility that these cells, through their trophic/regenerative potential, may also influence the course of chronic degenerative disorders and prevent cartilage degradation in osteoarthritis.
Human Dental Pulp Stem Cells Hook into Biocoral Scaffold Forming an Engineered Biocomplex
Carlo Mango1', Francesca Paino2', Riccardo d' Aquino2 , Alfredo De Rosa3 , Giovanna lezzi4 , Adriano
Piattello4, Luigi Laino3, Thimios Mitsiadis5 , Vincenzo Desiderio2 , Francesco Mangano1, Gianpalolo
Papaccio2*, Virginia Tirino2
The aim of this study was to evaluate the behavior of human Dental Pulp Stem Cells (DPSCs), as well as human osteoblasts, when challenged on a Biocoral scaffold, which is a porous natural hydroxyapatite. For this purpose, human DPSCs were seeded onto a three-dimensional (3D) Biocoral Scaffold or on flask surface (control). Either normal or rotative (3D) cultures were performed. Scanning electron microscopic analyses, at 8, 24 and 48 h of culture showed that cells did not adhere on the external surface, but moved into the cavities inside the Biocoral structure. After 7, 15 and 30 days of culture, morphological and molecular analyses suggested that the Biocoral Structure. After 7, 15 and 30 days of culture, morphological and molecular analyses suggested that the Biocoral scaffold leads DPSCs to hook into the cavities where there cells quickly start to secrete the extra cellular matrix (ECM) and differentiate into osteoblasts. Controls human osteoblasts also moved into the internal cavities where they secreted the ECM. Histological sections revealed a diffuse bone formation inside the Biocoral samples seeded with DPSCs or human osteoblasts, where the original scaffold and the new secreted biomaterial were completely integrated and cells were found within the remaining cavities. In addition, RT-PCR analyses showed a significant increase of osteoblasts-related gene expression and above all, of those genes highly expressed in mineralized tissues, including osteocalcin, OPN and BSP. Furthermore, the effects on the interaction between osteogenesis and angiogenesis were observed and substantiated by ELISA assays. Taken together, our results provide clear evidence that DPSCs differentiated into osteoblasts, forming a biocomplex made of Biocoral, ECM and differentiated cells.
Dental Pulp Stem Cells and Tissue Engineering Strategies for Clinical Application on Odontoiatric Field
Zavan Barbara et al. University of Padova Italy
The dental pulp plays a major role in tooth regeneration after injury, participating process called reparative dentinogenesis.
Human dental pulp stem cell is a promising autologous seed cell for bone tissue engineering
LI Jing-hui, LIU Da-yong, ZHANG Fang-ming, WANG Fan, ZHANG Wen-kui and ZHANG Zhen-ting
Chin Med J 2011;124(23):4022-4028
In vitro studies revealed that hDPSCs do possess osteogenic differentiation potential .In vivo studies revealed that hDPSCs seeded on gelatin scaffolds can form bone structure in heterotopic sites of nude mice.
These findings suggested that hDPSCs may be valuable as seed cells for bone tissue engineering .As a special stem cell source ,hDPSCs may be blaze a new path for bone tissue engineering.
Odontogenic Differentiation of Human Dental pulp stem cells stimulated by the Calcium Phosphate Porous Granules
Sunyoung Nam, Jong-Eun Won, Cheol-Hwan Kim, and Hae –Won Kim
Effects of three-dimensional (3D) calcium phosphate (Cap) porous granules on the growth and odontogenic diffentiation of human dental pulp stem cells (hDPSCs) were examined for dental tissue engineering .hDPSCs isolated from human dental pulps were cultured for 3-4 passages, and populated on porous granules .Cell growth on the culture dish showed on ongoing increase for up to 21 days, whereas the growth on the 3D granules decreased after 14 days. This reduction in proliferative potential on the 3D granules was more conspicuous under the osteogenic medium conditions, indicating that the 3D granules may alkaline phosphatase activity, up-regulation of odontoblast- specific genes, including dentin sialosphosprotein (DSPP) and dentin matrix protein 1(DMPI) by quantitative polymerase chain reaction , and greater level of dentin sialoprotein synthesis by western blot. Moreover, the cellular mineralization, as assessed by Alizarin red S and calcium quantification, was significantly higher in the 3D CaP granules than in the culture dish. Taken all, the 3D CaP porous granules should be useful for dental tissue engineering in combination with hDPSCs by providing favorable 3D substrate conditions for cell growth and odontogenic development.
Stem cell-based treatments for Type 1 diabetes mellitus:
bone marrow, embryonic, hepatic, pancreatic and induced pluripotent stem cells
K. J. Godfrey1, B. Mathew1, J. C. Bulman1 , O. Shah1 , S. Clement2 and G. l. Gallicano3
Type I diabetes mellitus-Characterized by the permanent destruction of insulin-secreting β-cells-is responsive to cell-based treatment that replace lost β-cell populations. The current gold standard of pancreas transplantation provides only temporary independence from exogenous insulin and is fraught with complications, including increased mortality. Stem cells offer a number of theoretical advantages over current therapies. Our review will focus on the development of treatments involving tissue stem cells from bone marrow, liver and pancreatic cells, as well as the potential use of embryonic and included pluripotent stem cells for Type 1 diabetes therapy. These cells have been demonstrated to increase endogenous insulin production, while partially mitigating the autoimmune destruction newly formed β-cells.
Diabet. Med. 29, 14-23 (2012)
Sinus Lift Augmentation Using Autologous Pulp Stem Cells: Case Report of Bone Density Evaluation
EUROPEAN JOURNAL OF INFLAMMATION
G. BRUNELLI, A. MOTRONI, F. CARINCI, A. GRAZIANO, R. D'AQUINO, I. ZOLLINO, R. MONGUZZI
Don Orione Hospital, Bergamo, Italy
(Applied Medical Imaging Research Group), Milan, Italy
Department of D.M.C.C.,Section of Maxillofacial and Plastic Surgery, University of Ferrara, Ferrara, Italy
Bone augmentation to reconstruct atrophic jaws provides the base for sufficient functional and aesthetic implant-supported oral rehabilitation. Although autografts are the standard procedure for bone grafting, the use of bone regeneration by means of dental pulp stem cell is an alternative that open a new era in this field. In March 2010, at the Department of Oral Surgery, Don Orione Hospital, Bergamo, Italy, one patient undergo to sinus lift elevation with pulp stem cells gentle poured onto collagen sponge. A CT scan control was performed after 4 month and DICOM data were processed with medical imaging software which gives the possibility to use a virtual probe to extract the bone density. Pearson's chi-square test was used to investigate difference in bone density (i.e. BD) between native and newly formed bone. BD in newly formed bone is about the double of native bone. This report demonstrated that stem cells derived from dental pulp poured onto collagen sponge is a useful method for bone regeneration in atrophic maxilla.
Human CD34+ Stem Cells Produce Bone Nodules in Vivo
Cell Prolif.2008, 41
A. Graziano, R. d'Aquino, G. Laino, A. Proto, M. T. Giuliano, G. Pirozzi, A. De Rosa, D. Di Napoli and G. Papaccio
Dipartimento di Medicina Sperimentale, Sezione Istologia ed Embriologia, TESLab, Secondo Ateneo di Napoli, Napoli, Italy, Dipartimento di Discipline Odontostomatologiche, Ortodontiche e Chirurgiche, Secondo Ateneo di Napoli, Napoli, Italy, Dipartimento di Chimica Università degli Studi di Salerno, Salerno, Italy, Dipartimento di Medicina Sperimentale, Sezione di Biotecnologie, Secondo Ateneo di Napoli, Napoli, Italy, Oncologia Sperimentale C-Immunologia, Istituto Nazionale Tumori, Napoli, Italy, and Centro di Biotecnologie, AORN Cardarelli, Napoli, Italy
Objectives: The aim of this study was to select and provide enough stem cells for quick transplantation in bone engineering procedures.
Materials and Methods: germ pulp, collected from 25 healthy subjects aged 13–20 years, were subjected to magnetic-activated cell sorting to select a CD34+ Stem cell population capable of differentiating into pre-osteoblasts. These cells were allowed to adhere to an absorbable polylactic–coglycolic acid scaffold for 30 min, without any prior expansion, and the CD34+ cell-colonized scaffolds were then transplanted into immunocompromised rats, subcutaneously.
Results: After 60 days, analysis of recovered transplants revealed that they were formed of nodules of bone, of the same dimensions as the original scaffold.
This study indicates that CD34+cells obtained from dental pulp can be used for engineering bone, without the need for prior culture expanding procedures. Using autologous stem cells, this schedule could be used to provide the basis for bone regenerative surgery, with limited sacrifice of tissue, low morbidity at the collection site, and significant reduction in time needed for clinical recovery.
Human Dental Pulp Stem Cells Improve Left Ventricular Function, Induce Angiogenesis, and Reduce Infarct Size in Rats with Acute Myocardial Infarction
Unidad de Cardiorregeneracio´n, Centro de Investigacio´n Prı´ncipe Felipe, Valencia, Spain
Human dental pulp contains precursor cells termed dental pulp stem cells (DPSC) that show self-renewal and multilineage differentiation and also secrete multiple proangiogenic and antiapoptotic factors. To examine whether these cells could have therapeutic potential in the repair of myocardial infarction (MI), DPSC were infected with a retrovirus encoding the green fluorescent protein (GFP) and expanded ex vivo. Seven days after induction of myocardial infarction by coronary artery ligation, 1.5 _ 106 GFP-DPSC were injected intramyocardially in nude rats. At 4 weeks, cell-treated animals showed an improvement in cardiac function, observed by percentage changes in anterior wall thickening left ventricular fractional area change, in parallel with a reduction in infarct size. No histologic evidence was seen of GFP+ endothelial cells, smooth muscle cells, or cardiac muscle cells within the infarct. However, angiogenesis was increased relative to control-treated animals. Taken together, these data suggest that DPSC could provide a novel alternative cell population for cardiac repair, at least in the setting of acute MI. STEM CELLS 2008;26:638–645
Human Postnatal Dental Pulp Cells Co-Differentiate into Osteoblasts and Endotheliocytes: A Pivotal Synergy Leading to Adult Bone Tissue Formation
Cell Death and Differentiation (2007) 14
R d'Aquino, M Sampaolesi, G Laino, G Pirozzi, A De Rosa and G Papaccio
Dipartimento di Discipline Odontostomatologiche, Ortodontiche e Chirurgiche, Universita` Secondo Ateneo di Napoli, Napoli, Italy; Dipartimento di Medicina Sperimentale, Sezione di Istologia ed Embriologia, Universita` Secondo Ateneo di Napoli, Napoli, Italy; DIBIT Dipartimentale San Raffaele Scientific Institute, Universita` Vita e Salute, Stem Cell Research Institute, Milan, Italy and UOS Dipartimentale, Diagnostica Molecolare, Analisi d'immagine e citometrı´a a flusso, Istituto Nazionale Tumori 'G. Pascale', Napoli, Italy
Stromal stem cells from human dental pulp (SBP-DPSCs) were used to study osteogenic differentiation in vitro and in vivo. We previously reported that SBP-DPSCs are multipotent stem cells able to differentiate into osteoblasts, which synthesize three dimensional woven bone tissue chips in vitro. In this study, we followed the temporal expression pattern of specific markers in SBP-DPSCs and found that, when differentiating into osteoblasts, they express, besides osteocalcin, also flk-1 (VEGF-R2). In addition, 30% of them expressed specific antigens for endothelial cells, including CD54, von-Willebrand (domain 1 and 2), CD31 (PECAM-1) and angiotensin-converting enzyme. Interestingly, we found endotheliocytes forming vessel walls, observing that stem cells synergically differentiate into osteoblasts and endotheliocytes, and that flk-1 exerts a pivotal role in coupling osteoblast and endotheliocyte differentiation. When either SBP-DPSCs or bone chips obtained in vitro were transplanted into immunocompromised rats, they generated a tissue structure with an integral blood supply similar to that of human adult bone; in fact, a large number of HLA-1 + vessels were observed either within the bone or surrounding it in a periosteal layer. This study provides direct evidence to suggest that osteogenesis and angiogenesis mediated by human SBP-DPSCs may be regulated by distinct mechanisms, leading to the organization of adult bone tissue after stem cell transplantion.
Cell Death and Differentiation (2007) 14,1162-1171.doi:10.1038/sj.cdd.4402121;published online 9 March 2007