In vitro histomorphometric comparison of dental pulp tissue in different teeth

Background Dental pulp (DP) represents an accessible and valuable source promising of stem cells for clinical application. However, there are some disadvantages associated with the isolation of dental pulp stem cells (DPSCs), which include the size and weight of the pulp tissue needed to yield sufficient cells for culturing in vitro. Therefore, the objective of this study was to compare in vitro histomorphometry of DP from permanent (premolars, third molar), supernumerary and deciduous teeth of patients between 5 and 25 years old with regards to weight, length, width and the cell density in the four regions of the DP in order to obtain quantitative parameters in a tissue that represents a valuable source of stem cells. Methods DPs were obtained from 10 central incisors deciduous, 20 permanent teeth (10 premolars, 10 third molars) and 10 supernumeraries (six mesiodents and four inferior premolar shapes). The pulps were carefully removed, and the entire tissue was weighed. The pulp length and the width were measured with a digital Vernier caliper. The cellular density analysis was performed according to the four regions of the DP (coronal, cervical, medial and apical) in histological slides using photography and the ImageJ® program for quantification. Results The Pearson correlation test revealed that DP weight among different types of teeth is correlated with age in male patients. A significant positive correlation was noted between length and width of the DP with age in both genders. The mean DP weight for supernumerary and third molar teeth was greater than deciduous and premolar teeth. Finally, the histological analysis showed that the coronal and apical portions of DP in supernumerary and premolar teeth have the highest cell density. Conclusions The DP of supernumerary teeth has quantitatively the best morphometric parameters and cell density comparable with the quality of DP obtained from deciduous teeth.

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Stem Cells Derived from Human Exfoliated Deciduous teeth [shed] in Neuronal Disorders: A Review

Current Stem Cell Research & Therapy ◽

10.2174/1574888x16666201221151512 ◽

2020 ◽

Vol 16 ◽

Author(s):

Minu Anoop ◽

Indrani Datta

Keyword(s):

Stem Cells ◽

Neurodegenerative Diseases ◽

Dental Pulp ◽

Therapeutic Potential ◽

Permanent Teeth ◽

Deciduous Teeth ◽

Proliferative Potential ◽

Pulp Tissue ◽

Human Exfoliated Deciduous Teeth

: Most conventional treatments for neurodegenerative diseases fail due to their focus on neuroprotection rather than neurorestoration. Stem cell‐based therapies are becoming a potential treatment option for neurodegenerative diseases as they can home in, engraft, differentiate and produce factors for CNS recovery. Stem cells derived from human dental pulp tissue differ from other sources of mesenchymal stem cells due to their embryonic neural crest origin and neurotrophic property. These include both dental pulp stem cells [DPSCs] from dental pulp tissues of human permanent teeth and stem cells from human exfoliated deciduous teeth [SHED]. SHED offer many advantages over other types of MSCs such as good proliferative potential, minimal invasive procurement, neuronal differentiation and neurotrophic capacity, and negligible ethical concerns. The therapeutic potential of SHED is attributed to the paracrine action of extracellularly released secreted factors, specifically the secretome, of which exosomes is a key component. SHED and its conditioned media can be effective in neurodegeneration through multiple mechanisms, including cell replacement, paracrine effects, angiogenesis, synaptogenesis, immunomodulation, and apoptosis inhibition, and SHED exosomes offer an ideal refined bed-to-bench formulation in neurodegenerative disorders. However, in spite of these advantages, there are still some limitations of SHED exosome therapy, such as the effectiveness of long-term storage of SHED and their exosomes, the development of a robust GMP-grade manufacturing protocol, optimization of the route of administration, and evaluation of the efficacy and safety in humans. In this review, we have addressed the isolation, collection and properties of SHED along with its therapeutic potential on in vitro and in vivo neuronal disorder models as evident from the published literature.

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Isolation and culture of dental pulp stem cells from permanent and deciduous teeth

Pakistan Journal of Medical Sciences ◽

10.12669/pjms.35.4.540 ◽

2019 ◽

Vol 35 (4) ◽

Cited By ~ 1

Author(s):

Shagufta Naz ◽

Farhan Raza Khan ◽

Raheela Rahmat Zohra ◽

Sahreena Salim Lakhundi ◽

Mehwish Sagheer Khan ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Permanent Teeth ◽

Deciduous Teeth ◽

Tooth Pulp ◽

Human Teeth ◽

Creative Commons ◽

Calcified Tissue

Objective: To isolate dental pulp mesenchymal stem cells (MSCs) from non-infected human permanent and deciduous teeth. Methods: It was an in-vitro experimental study. Human teeth were collected from 13 apparently healthy subjects including nine adults and four children. After decoronation dental pulps were extirpated from teeth and cultured via explant method in a stem cell defined media. Data was analyzed by descriptive statistics. Results: As above MSCs emerged exhibiting fibroblast-like morphology. In vitro culture was positive for 100% (9/9) and 75% (3/4) of the permanent and deciduous teeth respectively. First cell appeared from deciduous teeth pulp in 10±6.2 days while permanent teeth pulp took 12.4±3.7 days. Together, 26.6±3.6 and 24.5±3.5 days were required for permanent and deciduous tooth pulp stem cells to be ready for further assays. Conclusions: The protocol we developed is easy and consistent and can be used to generate reliable source of MScs for engineering of calcified and non-calcified tissue for regenerative medicine approaches. doi: https://doi.org/10.12669/pjms.35.4.540 How to cite this:Naz S, Khan FR, Zohra RR, Lakhundi SS, Khan MS, Mohammed N, et al. Isolation and culture of dental pulp stem cells from permanent and deciduous teeth. Pak J Med Sci. 2019;35(4):---------. doi: https://doi.org/10.12669/pjms.35.4.540 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Insight into the Role of Dental Pulp Stem Cells in Regenerative Therapy

Biology ◽

10.3390/biology9070160 ◽

2020 ◽

Vol 9 (7) ◽

pp. 160 ◽

Cited By ~ 5

Author(s):

Shinichiro Yoshida ◽

Atsushi Tomokiyo ◽

Daigaku Hasegawa ◽

Sayuri Hamano ◽

Hideki Sugii ◽

...

Keyword(s):

Stem Cells ◽

Tissue Regeneration ◽

Dental Pulp ◽

High Growth ◽

Dental Pulp Stem Cells ◽

Permanent Teeth ◽

Deciduous Teeth ◽

Pulp Tissue ◽

Differentiation Potential ◽

Cell Based Therapy

Mesenchymal stem cells (MSCs) have the capacity for self-renewal and multilineage differentiation potential, and are considered a promising cell population for cell-based therapy and tissue regeneration. MSCs are isolated from various organs including dental pulp, which originates from cranial neural crest-derived ectomesenchyme. Recently, dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHEDs) have been isolated from dental pulp tissue of adult permanent teeth and deciduous teeth, respectively. Because of their MSC-like characteristics such as high growth capacity, multipotency, expression of MSC-related markers, and immunomodulatory effects, they are suggested to be an important cell source for tissue regeneration. Here, we review the features of these cells, their potential to regenerate damaged tissues, and the recently acquired understanding of their potential for clinical application in regenerative medicine.

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VEGFR1 primes a unique cohort of dental pulp stem cells for vasculogenic differentiation

10.22203/ecm.v041a21 ◽

2021 ◽

Vol 41 ◽

pp. 332-344

Author(s):

MT Bergamo ◽

◽

Z Zhang ◽

TM Oliveira ◽

JE Nör

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Vascular Endothelial Cells ◽

Dental Pulp Stem Cells ◽

Deciduous Teeth ◽

Microvascular Endothelial Cell ◽

Vascular Endothelial ◽

Pulp Tissue ◽

Immunodeficient Mice

Dental pulp stem cells (DPSCs) constitute a unique group of cells endowed with multipotency, self-renewal, and capacity to regenerate the dental pulp tissue. While much has been learned about these cells in recent years, it is still unclear if each DPSC is multipotent or if unique sub-populations of DPSCs are “primed” to undergo specific differentiation paths. The purpose of the present study was to define whether a sub-population of DPSCs was uniquely primed to undergo vasculogenic differentiation. Permanent-tooth DPSCs or stem cells from human exfoliated deciduous teeth (SHED) were flow-sorted for vascular endothelial growth factor receptor 1 (VEGFR1) and exposed to vasculogenic differentiation medium, i.e., Microvascular-Endothelial-Cell-Growth-Medium-2-BulletKit™ supplemented with 50 ng/mL rhVEGF165 in the presence of 0 or 25 μg/mL anti-human VEGF antibody (bevacizumab; Genentech). In addition, sorted SHED (i.e., VEGFR1high or VEGFR1low) were seeded in biodegradable scaffolds and transplanted into the subcutaneous space of immunodeficient mice. Despite proliferating at a similar rate, VEGFR1high generated more in vitro sprouts than VEGFR1low cells (p < 0.05). Blockade of VEGF signaling with bevacizumab inhibited VEGFR1high-derived sprouts, demonstrating specificity of responses. Similarly, VEGFR1high SHED generated more blood vessels when transplanted into murine hosts than VEGFR1low cells (p < 0.05). Collectively, these data demonstrated that DPSCs contain a unique sub-population of cells defined by high VEGFR1 expression that are primed to differentiate into vascular endothelial cells. These data raise the possibility of purifying stem cells with high vasculogenic potential for regeneration of vascularized tissues or for vascular engineering in the treatment of ischemic conditions.

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Regenerative potential of stem cells derived from human exfoliated deciduous (SHED) teeth during engineering of human body tissues

Current Stem Cell Research & Therapy ◽

10.2174/1574888x16999201231213206 ◽

2020 ◽

Vol 16 ◽

Author(s):

Shivkanya Fuloria ◽

Ajay Jain ◽

Sameep Singh ◽

Iswar Hazarika ◽

Samson Salile ◽

...

Keyword(s):

Stem Cells ◽

Human Body ◽

In Vivo Studies ◽

Permanent Teeth ◽

Deciduous Teeth ◽

Pulp Tissue ◽

Human Teeth ◽

Body Tissues

Abstract:: Current decade witnesses the regenerative potential of stem cells (SCs) based life-saving therapies for the treatment of various disease and conditions. Human teeth act as reservoir for SCs that exist with high abundance in baby, wisdom, and permanent teeth. The collection of stem cells from human exfoliated deciduous teeth (SHED) is considered as a simple process as it offers the convenience of little or no pain. In comparison to the SCs from dental or bone marrow or other tissues, the SHED offers the benefit of higher cellular differentiation and proliferation. Massive in vitro and in vivo studies reveal the regenerative potential of SHED in the engineering of dental pulp tissue, neuronal tissue, root, bio root, cardiovascular tissues, lymphatic tissues, renal tissues, dermal tissues, hepatic tissues, and bone tissues. Current review describes the methods of collection/isolation/storage, various biomarkers, and types of SHED. This review highlights the regenerative potential of SHED in the engineering of different tissues of human body. As per the available research evidences present study supports that SHED may differentiate into the endothelial cells, neurons, odontoblasts, pancreatic

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Isolation and characterization of human dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED) – an in vitro study

RGUHS Journal of Dental Sciences ◽

10.26715/rjds.10_2_3 ◽

2018 ◽

Vol 10 (2) ◽

pp. 9-18

Author(s):

Dr. Vinay Rao ◽

Dr. Roopa R Nadig ◽

Dr.Ramanand Nadig ◽

Dr.Karthik .J ◽

Dr. Veena Pai S

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Surface Characterization ◽

Dental Pulp Stem Cells ◽

In Vivo Studies ◽

Deciduous Teeth ◽

Pulp Tissue ◽

Isolation And Characterization

AIMS AND OBJECTIVES: 1. Isolation and growth of dental pulp stem cells (DPSCs) and stem cells from exfoliated human deciduous teeth (SHED). 2. Characterization 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 the help of flow cytometer using a panel of various surface markers. An immuno cytochemistry analysis was done to see the expression of proteins in the cells. RESULTS: Identification of cells was done with the help of a phase contrast microscope. Flow cytometry analyses for various CD markers showed similar results for both DPSCs and SHED. The cells showed positive expression for pluripotent markers, ectodermal markers and mesodermal markers. CONCLUSION: The study demonstrated that stem cells existed in human deciduous and permanent pulp tissue. The stem cells present in deciduous permanent 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. Further studies should include analysis of diverse cell populations to elucidate their potential to differentiate into various cell types followed by in vivo studies in animals and humans.

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Comparative evaluation of osteogenic differentiation potential of stem cells derived from dental pulp and exfoliated deciduous teeth cultured over granular hydroxyapatite based scaffold

BMC Oral Health ◽

10.1186/s12903-021-01621-0 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Manal Nabil Hagar ◽

Farinawati Yazid ◽

Nur Atmaliya Luchman ◽

Shahrul Hisham Zainal Ariffin ◽

Rohaya Megat Abdul Wahab

Keyword(s):

Stem Cells ◽

Osteogenic Differentiation ◽

Dental Pulp ◽

3D Culture ◽

Permanent Teeth ◽

Deciduous Teeth ◽

Osteogenic Potential ◽

Control Group ◽

Rt Pcr ◽

Differentiation Potential

Abstract Background Mesenchymal stem cells isolated from the dental pulp of primary and permanent teeth can be differentiated into different cell types including osteoblasts. This study was conducted to compare the morphology and osteogenic potential of stem cells from exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSC) in granular hydroxyapatite scaffold (gHA). Preosteoblast cells (MC3T3-E1) were used as a control group. Methodology The expression of stemness markers for DPSC and SHED was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR). Alkaline phosphatase assay was used to compare the osteoblastic differentiation of these cells (2D culture). Then, cells were seeded on the scaffold and incubated for 21 days. Morphology assessment using field emission scanning electron microscopy (FESEM) was done while osteogenic differentiation was detected using ALP assay (3D culture). Results The morphology of cells was mononucleated, fibroblast-like shaped cells with extended cytoplasmic projection. In RT-PCR study, DPSC and SHED expressed GAPDH, CD73, CD105, and CD146 while negatively expressed CD11b, CD34 and CD45. FESEM results showed that by day 21, dental stem cells have a round like morphology which is the morphology of osteoblast as compared to day 7. The osteogenic potential using ALP assay was significantly increased (p < 0.01) in SHED as compared to DPSC and MC3T3-E1 in 2D and 3D cultures. Conclusion gHA scaffold is an optimal scaffold as it induced osteogenesis in vitro. Besides, SHED had the highest osteogenic potential making them a preferred candidate for tissue engineering in comparison with DPSC.

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Hypoxia-inducible factor 1α induces osteo/odontoblast differentiation of human dental pulp stem cells via Wnt/β-catenin transcriptional cofactor BCL9

Scientific Reports ◽

10.1038/s41598-021-04453-8 ◽

2022 ◽

Vol 12 (1) ◽

Author(s):

Shion Orikasa ◽

Nobuyuki Kawashima ◽

Kento Tazawa ◽

Kentaro Hashimoto ◽

Keisuke Sunada-Nara ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Target Genes ◽

Hypoxia Inducible Factor ◽

Dental Pulp Stem Cells ◽

Pulp Tissue ◽

Odontoblast Differentiation ◽

Hypoxia Inducible Factor 1Α ◽

Hypoxic Conditions

AbstractAccelerated dental pulp mineralization is a common complication in avulsed/luxated teeth, although the mechanisms underlying this remain unclear. We hypothesized that hypoxia due to vascular severance may induce osteo/odontoblast differentiation of dental pulp stem cells (DPSCs). This study examined the role of B-cell CLL/lymphoma 9 (BCL9), which is downstream of hypoxia-inducible factor 1α (HIF1α) and a Wnt/β-catenin transcriptional cofactor, in the osteo/odontoblastic differentiation of human DPSCs (hDPSCs) under hypoxic conditions. hDPSCs were isolated from extracted healthy wisdom teeth. Hypoxic conditions and HIF1α overexpression induced significant upregulation of mRNAs for osteo/odontoblast markers (RUNX2, ALP, OC), BCL9, and Wnt/β-catenin signaling target genes (AXIN2, TCF1) in hDPSCs. Overexpression and suppression of BCL9 in hDPSCs up- and downregulated, respectively, the mRNAs for AXIN2, TCF1, and the osteo/odontoblast markers. Hypoxic-cultured mouse pulp tissue explants showed the promotion of HIF1α, BCL9, and β-catenin expression and BCL9-β-catenin co-localization. In addition, BCL9 formed a complex with β-catenin in hDPSCs in vitro. This study demonstrated that hypoxia/HIF1α-induced osteo/odontoblast differentiation of hDPSCs was partially dependent on Wnt/β-catenin signaling, where BCL9 acted as a key mediator between HIF1α and Wnt/β-catenin signaling. These findings may reveal part of the mechanisms of dental pulp mineralization after traumatic dental injury.

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Serine Metabolism Controls Dental Pulp Stem Cell Aging by Regulating the DNA Methylation of p16

Journal of Dental Research ◽

10.1177/0022034520958374 ◽

2020 ◽

Vol 100 (1) ◽

pp. 90-97

Author(s):

R.L. Yang ◽

H.M. Huang ◽

C.S. Han ◽

S.J. Cui ◽

Y.K. Zhou ◽

...

Keyword(s):

Dna Methylation ◽

Stem Cells ◽

Stem Cell ◽

Dental Pulp ◽

Permanent Teeth ◽

Deciduous Teeth ◽

Cell Aging ◽

Differentiation Capacity ◽

Stem Cell Aging ◽

Serine Metabolism

To investigate the characteristics and molecular events of dental pulp stem cells (DPSCs) for tissue regeneration with aging, we isolated and analyzed the stem cells from human exfoliated deciduous teeth (SHED) and permanent teeth of young (Y-DPSCs) and old (A-DPSCs) adults. Results showed that the stemness and osteogenic differentiation capacity of DPSCs decreased with aging. The RNA sequencing results showed that glycine, serine, and threonine metabolism was one of the most enriched gene clusters among SHED, Y-DPSCs, and A-DPSCs, according to analysis based on the Kyoto Encyclopedia of Genes and Genomes. The expression of serine metabolism–related enzymes phosphoserine aminotransferase 1 (PSAT1) and phosphoglycerate (PHGDH) decreased in A-DPSCs and provided less methyl donor S-adenosylmethionine (SAM) for DNA methylation, leading to the hypomethylation of the senescence marker p16 (CDNK2A). Furthermore, the proliferation and differentiation capacity of Y-DPSCs and SHED decreased after PHGDH siRNA treatment, which reduced the level of SAM. Convincingly, the ratios of PSAT1-, PHGDH-, or proliferating cell nuclear antigen–positive cells in the dental pulp of old permanent teeth were less than those in the dental pulp of deciduous teeth and young permanent teeth. In summary, the stemness and differentiation capacity of DPSCs decreased with aging. The decreased serine metabolism in A-DPSCs upregulated the expression of p16 via attenuating its DNA methylation, resulting in DPSC aging. Our finding indicated that serine metabolism and 1 carbon unit participated in stem cell aging, which provided new direction for stem cell aging study and intervention.

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Therapeutic applications of stem cells from human exfoliated deciduous teeth: a review

International Journal of Research in Medical Sciences ◽

10.18203/2320-6012.ijrms20203705 ◽

2020 ◽

Vol 8 (9) ◽

pp. 3413

Author(s):

Mani Baweja

Keyword(s):

Stem Cells ◽

Animal Studies ◽

Permanent Teeth ◽

Deciduous Teeth ◽

Oral Diseases ◽

Dental Stem Cells ◽

Medline Search ◽

Therapeutic Applications ◽

Human Exfoliated Deciduous Teeth

Dental stem cells have been found to have the ability to differentiate into nerve cells, adipose cells, chondrocytes, osteoblasts, myocytes, hepatocytes, and odontoblasts. They can be derived from permanent teeth or deciduous teeth. Stem cells from human exfoliated deciduous teeth (SHED) have a higher proliferation rate and higher osteogenic and neurogenic potential than dental pulp stem cells (DPSC). Therefore, SHEDs are an attractive cell source for tissue regeneration. A large plethora of in vitro and animal studies have been conducted in the last few decades that has demonstrated the potential uses of these cells for the treatment of oral and non-oral diseases. The aim of this article was to review the potential therapeutic applications of stem cells derived from human exfoliated deciduous teeth. A Medline search was done, including international literature, published in English between 2003 and 2020. In this area, further research is needed to ensure the applicability of SHED in the treatment of diseases in humans.

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