Therapeutic Potential of Dental Pulp Stem Cells and Leukocyte- and Platelet-Rich Fibrin for Osteoarthritis

Osteoarthritis (OA) is a degenerative and inflammatory joint disorder with cartilage loss. Dental pulp stem cells (DPSCs) can undergo chondrogenic differentiation and secrete growth factors associated with tissue repair and immunomodulation. Leukocyte- and platelet-rich fibrin (L-PRF) emerges in regenerative medicine because of its growth factor content and fibrin matrix. This study evaluates the therapeutic application of DPSCs and L-PRF in OA via immunomodulation and cartilage regeneration. Chondrogenic differentiation of DPSCs, with or without L-PRF exudate (ex) and conditioned medium (CM), and of bone marrow-mesenchymal stem cells was compared. These cells showed differential chondrogenesis. L-PRF was unable to increase cartilage-associated components. Immature murine articular chondrocytes (iMACs) were cultured with L-PRF ex, L-PRF CM, or DPSC CM. L-PRF CM had pro-survival and proliferative effects on unstimulated and cytokine-stimulated iMACs. L-PRF CM stimulated the release of IL-6 and PGE2, and increased MMP-13, TIMP-1 and IL-6 mRNA levels in cytokine-stimulated iMACs. DPSC CM increased the survival and proliferation of unstimulated iMACs. In cytokine-stimulated iMACs, DPSC CM increased TIMP-1 gene expression, whereas it inhibited nitrite release in 3D culture. We showed promising effects of DPSCs in an in vitro OA model, as they undergo chondrogenesis in vitro, stimulate the survival of chondrocytes and have immunomodulatory effects.

Download Full-text

Novel nanocomposite scaffold based on Gelatin/PLGA-PEG-PLGA hydrogels embedded with TGF-β1 for chondrogenic differentiation of human dental pulp stem cells in vitro

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2021.12.097 ◽

2022 ◽

Author(s):

Parisa Ghandforoushan ◽

Jalal Hanaee ◽

Zahra Aghazadeh ◽

Mohammad Samiei ◽

Amir Mohammad Navali ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Chondrogenic Differentiation ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp ◽

Nanocomposite Scaffold ◽

Tgf Β1

Download Full-text

In Vitro Characterization of Dental Pulp Stem Cells Cultured in Two Microsphere-Forming Culture Plates

Journal of Clinical Medicine ◽

10.3390/jcm9010242 ◽

2020 ◽

Vol 9 (1) ◽

pp. 242 ◽

Cited By ~ 2

Author(s):

Nam-Ung Bu ◽

Hyo-Seol Lee ◽

Bin-Na Lee ◽

Yun-Chan Hwang ◽

Sun-Young Kim ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Differentially Expressed Gene ◽

3D Culture ◽

Dental Pulp Stem Cells ◽

Rna Seq ◽

Multilineage Differentiation ◽

Culture Methods ◽

Differentiation Potential

Various three-dimensional (3D) culture methods have been introduced to overcome the limitations of in vitro culture and mimic in vivo conditions. This study aimed to evaluate two microsphere-forming culture methods and a monolayer culture method. We evaluated cell morphology, viability, osteo-, adipo-, and chondrogenic differentiation potential of dental pulp stem cells (DPSCs) cultured in 3D culture plates: ultra-low attachment (ULA) and U-bottomed StemFit 3D (SF) plates, and a two-dimensional (2D) monolayer plate. RNA sequencing (RNA-seq) revealed differentially expressed gene (DEG) profiles of the DPSCs. In contrast to an increasing pattern in the 2D group, cell viability in 3D groups (ULA and SF) showed a decreasing pattern; however, high multilineage differentiation was observed in both the 3D groups. RNA-seq showed significantly overexpressed gene ontology categories including angiogenesis, cell migration, differentiation, and proliferation in the 3D groups. Hierarchical clustering analysis revealed a similar DEG regulation pattern between the 3D groups; however, a comparatively different DEG was observed between the 2D and 3D groups. Taken together, this study shows that DPSCs cultured in microsphere-forming plates present superior multilineage differentiation capacities and demonstrate higher DEG expression in regeneration-related gene categories compared to that in DPSCs cultured in a conventional monolayer plate.

Download Full-text

Increased VEGF-A Expression of Human Dental Pulp Stem Cells (hDPSCs) Cultured with Advanced Platelet Rich Fibrin (A-PRF)

The Open Dentistry Journal ◽

10.2174/1874210602115010569 ◽

2021 ◽

Vol 15 (1) ◽

pp. 569-574

Author(s):

Dini Asrianti Bagio ◽

Indah Julianto ◽

Anggraini Margono ◽

Endang Suprastiwi

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Control Group ◽

Platelet Rich Fibrin ◽

Significant Difference ◽

Angiogenesis Process ◽

Human Dental Pulp ◽

Post Hoc

Background: VEGF-A expression of human dental pulp stem cells (hDPSCs) can induce the angiogenesis process of dental pulp regeneration. This in vitro study aimed to analyze the effect of various concentrations of Advanced Platelet Rich Fibrin (A-PRF) conditioned media (CM) on the increased expression of vascular endothelial growth factor-A (VEGF-A) of hDPSCs. Methods: hDPSCs were collected from ten third molars extracted from nine healthy donors, cultured, and then harvested at the end of the third passage. The hDPSCs were seeded in four different CM (control group: hDPSCs + DMEM; 1% A-PRF CM group: hDPSCs + 1% A-PRF CM; 5% A-PRF CM group: hDPSCs + 5% A-PRF CM; 10% A-PRF CM group: hDPSCs + 10% A-PRF CM). All of the groups were cultured in biological triplicates (Triplo) and observed for 5, 12, and 24 hours. The VEGF-A protein expression of hDPSCs was measured using human VEGF-A ELISA at a wavelength of 405 nm. Data was analyzed with Kruskal Wallis and post hoc Mann Whitney test with p<0.05. Results: The VEGF-A expression rate of hDPSCs among all groups was statistically significantly different at 5, 12 and 24 hours of observations (p<0.05). Post hoc analysis test showed a statistically significant difference of hDPSCs’s VEGF-A expression between 5% A-PRF groups compared to other groups at 5 and 12 hours of observation (p<0.05). However, there were no statistically significant differences observed of hDPSCs’ VEGF-A expression at 24 hours of observation between 1%, 5% and 10% A-PRF groups (p>0.05). Conclusion: 5% A-PRF CM was superior in increasing VEGF-A expression of hDPSCs at 5, 12 and 24 hours of observations.

Download Full-text

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.

Download Full-text

Modulation of Cell Death and Promotion of Chondrogenic Differentiation by Fas/FasL in Human Dental Pulp Stem Cells (hDPSCs)

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2020.00279 ◽

2020 ◽

Vol 8 ◽

Cited By ~ 1

Author(s):

Alessandra Pisciotta ◽

Giulia Bertani ◽

Laura Bertoni ◽

Rosanna Di Tinco ◽

Sara De Biasi ◽

...

Keyword(s):

Stem Cells ◽

Cell Death ◽

Dental Pulp ◽

Chondrogenic Differentiation ◽

Dental Pulp Stem Cells ◽

Human Dental Pulp

Download Full-text

Similar Features, Different Behaviors: A Comparative In Vitro Study of the Adipogenic Potential of Stem Cells from Human Follicle, Dental Pulp, and Periodontal Ligament

Journal of Personalized Medicine ◽

10.3390/jpm11080738 ◽

2021 ◽

Vol 11 (8) ◽

pp. 738

Author(s):

Melissa D. Mercado-Rubio ◽

Erick Pérez-Argueta ◽

Alejandro Zepeda-Pedreguera ◽

Fernando J. Aguilar-Ayala ◽

Ricardo Peñaloza-Cuevas ◽

...

Keyword(s):

Stem Cells ◽

Dental Pulp ◽

Periodontal Ligament ◽

Adipogenic Differentiation ◽

In Vitro Study ◽

Mrna Levels ◽

Dental Tissue ◽

Periodontal Ligament Stem Cells ◽

Differentiation Potential

Dental tissue-derived mesenchymal stem cells (DT-MSCs) are a promising resource for tissue regeneration due to their multilineage potential. Despite accumulating data regarding the biology and differentiation potential of DT-MSCs, few studies have investigated their adipogenic capacity. In this study, we have investigated the mesenchymal features of dental pulp stem cells (DPSCs), as well as the in vitro effects of different adipogenic media on these cells, and compared them to those of periodontal ligament stem cells (PLSCs) and dental follicle stem cells (DFSCs). DFSC, PLSCs, and DPSCs exhibit similar morphology and proliferation capacity, but they differ in their self-renewal ability and expression of stemness markers (e.g OCT4 and c-MYC). Interestingly, DFSCs and PLSCs exhibited more lipid accumulation than DPSCs when induced to adipogenic differentiation. In addition, the mRNA levels of adipogenic markers (PPAR, LPL, and ADIPOQ) were significantly higher in DFSCs and PLSCs than in DPSCs, which could be related to the differences in the adipogenic commitment in those cells. These findings reveal that the adipogenic capacity differ among DT-MSCs, features that might be advantageous to increasing our understanding about the developmental origins and regulation of adipogenic commitment.

Download Full-text

Enhanced Osteogenesis of Dental Pulp Stem Cells In Vitro Induced by Chitosan–PEG-Incorporated Calcium Phosphate Cement

Polymers ◽

10.3390/polym13142252 ◽

2021 ◽

Vol 13 (14) ◽

pp. 2252

Author(s):

Jae Eun Kim ◽

Sangbae Park ◽

Woong-Sup Lee ◽

Jinsub Han ◽

Jae Woon Lim ◽

...

Keyword(s):

Stem Cells ◽

Calcium Phosphate ◽

Zeta Potential ◽

Mechanical Strength ◽

Calcium Phosphate Cement ◽

Dental Pulp ◽

Dental Pulp Stem Cells ◽

Ion Trapping ◽

Alizarin Red

The use of bone graft materials is required for the treatment of bone defects damaged beyond the critical defect; therefore, injectable calcium phosphate cement (CPC) is actively used after surgery. The application of various polymers to improve injectability, mechanical strength, and biological function of injection-type CPC is encouraged. We previously developed a chitosan–PEG conjugate (CS/PEG) by a sulfur (VI) fluoride exchange reaction, and the resulting chitosan derivative showed high solubility at a neutral pH. We have demonstrated the CPC incorporated with a poly (ethylene glycol) (PEG)-grafted chitosan (CS/PEG) and developed CS/PEG CPC. The characterization of CS/PEG CPC was conducted using Fourier transform infrared spectroscopy (FT-IR) and X-ray diffraction (XRD). The initial properties of CS/PEG CPCs, such as the pH, porosity, mechanical strength, zeta potential, and in vitro biocompatibility using the WST-1 assay, were also investigated. Moreover, osteocompatibility of CS/PEG CPCs was carried out via Alizarin Red S staining, immunocytochemistry, and Western blot analysis. CS/PEG CPC has enhanced mechanical strength compared to CPC, and the cohesion test also demonstrated in vivo stability. Furthermore, we determined whether CS/PEG CPC is a suitable candidate for promoting the osteogenic ability of Dental Pulp Stem Cells (DPSC). The elution of CS/PEG CPC entraps more calcium ion than CPC, as confirmed through the zeta potential test. Accordingly, the ion trapping effect of CS/PEG is considered to have played a role in promoting osteogenic differentiation of DPSCs. The results strongly suggested that CS/PEG could be used as suitable additives for improving osteogenic induction of bone substitute materials.

Download Full-text