Search

Rat dental pulp stem cells (DPSCs) can be used to elucidate mesenchymal stem cell (MSC) applications in regenerative medicine. However, information on rat DPSCs during long-term passage, which could lead to replicative senescence, is limited. In this study, we investigated the phenotypic changes in DPSCs after 3–26 passages (3P–26P). The results show that cell morphology and nuclear size increase proportionally with passage number. The phosphorylated histone H2A.X (γ-H2A.X) positive cells (indicating DNA damage) increased significantly earlier than the 4-Hydroxynonenal (4-HNE) stained cells (indicating an abundance of intracellular reactive oxygen species). Compared to the cells subjected to 3P and 5P, the cells subjected to 15P showed reduced proliferation despite being positive for Ki67. Furthermore, cell growth was completely arrested after 26P. The senescence markers, senescence-associated β-galactosidase (SA-β-gal) and p16, exhibited similar expression patterns that were not correlated with those of p21 and urokinase-type plasminogen activator receptor (uPAR). Nearly all cells expressed SA-β-gal and p16 after 26P, whereas only half expressed p21 and uPAR. These results will contribute to understanding the characteristics of DPSCs toward replicative senescence, which are applicable to elucidate mechanisms related to regenerative medicine and stem cell aging. [ABSTRACT FROM AUTHOR]

BACKGROUND: Dental pulp stem cells are one of the stem cells with great potential in oral and maxillofacial tissue engineering. Compared with mesenchymal stem cells, dental pulp stem cells have the advantages of convenient collection, less ethical problems and higher potential of proliferation and differentiation. Currently, except for biochemical factors, physical stimulation also plays a critical role in the osteogenic/odontogenic differentiation of dental pulp stem cells. OBJECTIVE: To review the relevant physical factors and the possible signaling pathway affecting the osteogenic/odontogenic differentiation of dental pulp stem cells to find the optimal induction conditions affecting their differentiation. METHODS: PubMed and CNKI databases were searched for relevant articles using “dental pulp stem cells (DPSCs), osteogenesis differentiation, odontoblastic differentiation, hypoxia, mechanical force, laser therapy, magnetic fields, microgravity” as English and Chinese search terms. Seventy-nine articles regarding physical factors affecting osteogenic/odontogenic differentiation of dental pulp stem cells were selected for the review. RESULTS AND CONCLUSION: Direct or indirect physical signals in the microenvironment have shown broad application prospects in regulating the directed differentiation of stem cells. Many related physical factors, for example, hypoxia, mechanical stimulation (dynamic hydrostatic pressure, mechanical tension, shear force, etc.), laser, microgravity, and magnetic field, have positive influences on the osteogenic/odontogenic differentiation of dental pulp stem cells. Owing to the complex mechanical environment of stomatognathic system, mechanical stimulation is a key physical factor in changing cellular environment and is also a frontier in tissue engineering. It will provide new ideas for investigating the response of dental pulp stem cells to the mechanical environment in the diagnosis and treatment of oral diseases. Because this field is relatively “young”, the parameters of equipment have not been unified and the relevant results are not consistent. The optimal induction parameters and conditions of related physical factors should be further explored and optimized. Scaffold material, one of the three elements of tissue engineering, plays a role in promoting the osteogenic/odontogenic differentiation of dental pulp stem cells, and promotes the development of materials science and clinical technology. The signaling pathways involve Notch, Wnt, MAPK, etc. The biological basis of regulating the behavior of dental pulp stem cells is not clear. The specific mechanism will be further explored in the future to provide new ideas for dental pulp regeneration and bone tissue engineering under the influence of physical factors. [ABSTRACT FROM AUTHOR]

Abstract Background Differentiating dental pulp stem cells (DPSCs) into odontoblasts is a critical process for tooth self-repair and dentine‒pulp engineering strategies in the clinic. However, the mechanism underlying the regulation of DPSC odontoblastic differentiation remains largely unknown. Here, we demonstrated that BCL-2 interacting protein 3 (BNIP3)-dependent mitophagy is associated with importin subunit beta-1 (KPNB1)-activating transcription factor 4 (ATF4), which promotes DPSC odontoblastic differentiation. Methods The key genes involved in DPSC odontogenic differentiation were identified via bioinformatics. Stable silencing or overexpression of BNIP3 was performed to investigate its impact on DPSC differentiation in vitro (n ≥ 3). To explore the role of BNIP3 in vivo, tooth root fragments loaded with the hydrogel-transfected DPSC complex were implanted into nude mice (n ≥ 6). Dual-luciferase reporter assays and chromatin immunoprecipitation (ChIP) polymerase chain reaction (PCR) were conducted to explore the binding site of ATF4 to the BNIP3 promoter (n ≥ 3). Mitochondrial function experiments were performed to investigate the impact of ATF4-BNIP3 on mitochondria (n ≥ 3). Immunoprecipitation (IP) mass spectrometry (MS) was used to investigate the interaction between ATF4 and its binding protein, KPNB1. Plasmids containing wild-type (WT)/mutant (MUT)-nuclear localization signal (NLS) forms of ATF4 were constructed to determine the specific amino acid residues recognized by KPNB1 and their effects on DPSC odontoblastic differentiation (n ≥ 3). Results Compared with those in the control group, the levels of autophagy and mitophagy, especially BNIP3-dependent mitophagy, were greater in the DPSC odontoblastic differentiation group (P

Parkinson′s disease (PD) is the second most common neurodegenerative disease, mainly due to the loss of dopaminergic neurons in the substantia nigra (SN) of the midbrain, resulting in motor dysfunction. Dental stem cells (DSCs) are derived from the cranial neural crest which can be easily harvested. DSCs coming from the cranial neural crest have excellent proliferation and differentiation ability, and can promote nerve repair and regeneration. DSCs can secrete new dopamine neurons and secrete a large number of neurotrophic factors(NTFs) to improve the neural function, and also inhibit the neuroinflammatory response through immune modulation. DSCs can effectively alleviate the motor dysfunction in rats with PD and play an important role in R&D of regenerative medicine.

Background: Differentiating dental pulp stem cells (DPSCs) into odontoblasts is a critical process for tooth self-repair and dentine‒pulp engineering strategies in the clinic. However, the mechanism underlying the regulation of DPSC odontoblastic differentiation remains largely unknown. Here, we demonstrated that BCL-2 interacting protein 3 (BNIP3)-dependent mitophagy is associated with importin subunit beta-1 (KPNB1)-activating transcription factor 4 (ATF4), which promotes DPSC odontoblastic differentiation. Methods: The key genes involved in DPSC odontogenic differentiation were identified via bioinformatics. Stable silencing or overexpression of BNIP3 was performed to investigate its impact on DPSC differentiation in vitro (n ≥ 3). To explore the role of BNIP3 in vivo, tooth root fragments loaded with the hydrogel-transfected DPSC complex were implanted into nude mice (n ≥ 6). Dual-luciferase reporter assays and chromatin immunoprecipitation (ChIP) polymerase chain reaction (PCR) were conducted to explore the binding site of ATF4 to the BNIP3 promoter (n ≥ 3). Mitochondrial function experiments were performed to investigate the impact of ATF4-BNIP3 on mitochondria (n ≥ 3). Immunoprecipitation (IP) mass spectrometry (MS) was used to investigate the interaction between ATF4 and its binding protein, KPNB1. Plasmids containing wild-type (WT)/mutant (MUT)-nuclear localization signal (NLS) forms of ATF4 were constructed to determine the specific amino acid residues recognized by KPNB1 and their effects on DPSC odontoblastic differentiation (n ≥ 3). Results: Compared with those in the control group, the levels of autophagy and mitophagy, especially BNIP3-dependent mitophagy, were greater in the DPSC odontoblastic differentiation group (P < 0.05). Genetic silencing or overexpression of BNIP3 demonstrated that BNIP3 expression was positively correlated with the transition of DPSCs into odontoblasts both in vitro and in vivo (P < 0.05). ATF4 regulates the expression of BNIP3 by directly binding to approximately −1292 to −1279 bp and approximately −1185 to −1172 bp within the BNIP3 promoter region, which is associated with mitophagy and mitochondrial reactive oxygen species (mtROS) levels (P < 0.05). Moreover, ATF4 increased mitophagy, mitochondrial function, and cell differentiation potential via BNIP3 (P < 0.05). Mechanistically, KPNB1 is a novel interacting protein of ATF4 that specifically recognizes amino acids (aa) 280–299 within ATF4 to control its translocation into the nucleus and subsequent transcription and differentiation processes (P < 0.05). Conclusions: We reported that the critical role of KPNB1/ATF4/BNIP3 axis-dependent mitophagy could provide new cues for the regeneration of the dental pulp‒dentin complex in DPSCs. [ABSTRACT FROM AUTHOR]

The transplantation of collagen hydrogels encapsulating human dental pulp stem cell (DPSC)‐derived chondrogenic cells is potentially a novel approach for the regeneration of degenerated nucleus pulposus (NP) and cartilage. Grafted cell migration allows cells to disperse in the hydrogels and the treated tissue from the grafted location. We previously reported the cell migration in type I and type II hydrogels. It is important to explore further how cell culture time affect the cell motility. In this study, we observed the decreased motility of DPSC‐derived chondrogenic cells after culturing for 2 weeks compared with cells cultured for 2 days in these gels. The Alamarblue assay showed the cell proliferation during the two‐week cell culture period. The findings suggest that the transitions of cell motility and proliferation during the longer culture time. The result indicates that the early culture stage is an optimal time for cell transplantation. In a degenerated disc, the expression of IL‐1β and TNFα increased significantly compared with healthy tissue and therefore may affect grafted cell migration. The incorporation of IL‐1β and TNFα into the collagen hydrogels decreased cell motility. The study indicates that the control of IL‐1β and TNFα production may help to maintain cell motility after transplantation. [ABSTRACT FROM AUTHOR]

Secretome Dental Pulp Stem Cells have been proposed as a new alternative for wound burn. The aim of this study is to compare Secretome Dental Pulp Stem Cells and MEBO ointment against several fibroblasts and collagen in the healing process of superficial dermal burns. This study used a post-test only controlled group design by using 15 rats that were given superficial dermal burned wounds. Rats were divided into a negative control group of NaCl 0.9% (K1), a treatment group of MEBO (K2), and a treatment group of DPSCs-secretome (K3). Histopathology was done on the 5th, 14th, and 21st days post-induction wound burn. Network-prepared histopathology by colouring Messon Trichomes. Then, it was observed under a microscope with 200x magnification. Scoring in a way semi-qualitative for assessing fibroblasts and collagen. The total score of fibroblasts in the MEBO group was higher than the DPSCs-secretome group on day 5th. In comparison, the score for increasing the amount of collagen in the DPSCs-secretome group on days 5th and 14th was higher than the MEBO group. DPSCs-secretome has better effectiveness in increasing the amount of collagen but not better in increasing fibroblasts compared to MEBO. [ABSTRACT FROM AUTHOR]

Copyright of Basic & Clinical Medicine is the property of Editorial Office of Basic & Clinical Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Abstract Background Subarachnoid hemorrhage (SAH) is a severe stroke subtype that lacks effective treatment. Exosomes derived from human dental pulp stem cells (DPSCs) are a promising acellular therapeutic strategy for neurological diseases. However, the therapeutic effects of DPSC-derived exosomes (DPSC-Exos) on SAH remain unknown. In this study, we investigated the therapeutic effects and mechanisms of action of DPSC-Exos in SAH. Materials and methods SAH was established using 120 male Sprague-Dawley rats. One hour after SAH induction, DPSC-Exos were administered via tail vein injection. To investigate the effect of DPSC-Exos, SAH grading, short-term and long-term neurobehavioral assessments, brain water content, western blot (WB), immunofluorescence staining, Nissl staining, and HE staining were performed. The role of miR-197-3p/FOXO3 in regulating pyroptosis was demonstrated through miRNA sequencing, bioinformatics analysis, and rescue experiments. The SAH model in vitro was established by stimulating BV2 cells with hemoglobin (Hb) and the underlying mechanism of DPSC-Exos was investigated through WB and Hoechst/PI staining. Results The expressions of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) were increased after SAH. DPSC-Exos alleviated brain edema and neuroinflammation by inhibiting the expression of FOXO3 and reducing NLRP3 inflammasome activation, leading to improved neurobehavioral functions at 24 h after SAH. In vitro, the expression of the NLRP3 inflammasome components (NLRP3 and caspase1-p20), GSDMD-N, and IL-18 was inhibited in BV2 cells pretreated with DPSC-Exos. Importantly, DPSC-Exos overexpressing miR-197-3p had a more obvious protective effect than those from NC-transfected DPSCs, while those from DPSCs transfected with the miR-197-3p inhibitor had a weaker protective effect. Functional studies indicated that miR-197-3p bound to the 3ʹ-untranslated region of FOXO3, inhibiting its transcription. Furthermore, the overexpression of FOXO3 reversed the protective effects of miR-197-3p. Conclusions DPSC-Exos inhibited activation of the NLRP3 inflammasome and related cytokine release via the miR-197-3p/FOXO3 pathway, alleviated neuroinflammation, and inhibited microglial pyroptosis. These findings suggest that using DPSC-Exos is a promising therapeutic strategy for SAH. Graphical Abstract

Simple Summary: When joints are harmed due to injury, inheritance, or aging, it is difficult for the body to heal damaged cartilage. This is a major cause of pain and disability worldwide. With increasing life expectancy, millions of adults are affected, and enormous costs are imposed on health care systems. Scientists are attempting to use stem cells to create new cartilage-like tissue as a potential solution. However, the most suitable stem cell type for cartilage repair has not been defined. This study compared the ability to generate cartilage-like tissue by using two stem cell types that were derived from human bone marrow and dental pulp. To produce cartilage, the tissue's environment should be simulated for stem cells in a laboratory. Therefore, the low-oxygen tension of joint cartilage was simulated using a simple and inexpensive chemical, cobalt chloride. The results showed that dental pulp stem cells could produce higher quality tissue that closely resembled human joint cartilage in terms of its structure and organization that could form the primary base of normal function. While further research is necessary, these initial findings represent a promising step toward using stem cells more beneficially to enhance a patient's quality of life. The limited self-repair capacity of articular cartilage is a challenge for healing injuries. While mesenchymal stem/stromal cells (MSCs) are a promising approach for tissue regeneration, the criteria for selecting a suitable cell source remain undefined. To propose a molecular criterion, dental pulp stem cells (DPSCs) with a Hox-negative expression pattern and bone marrow mesenchymal stromal cells (BMSCs), which actively express Hox genes, were differentiated towards chondrocytes in 3D pellets, employing a two-step protocol. The MSCs' response to preconditioning by cobalt chloride (CoCl2), a hypoxia-mimicking agent, was explored in an assessment of the chondrogenic differentiation's efficiency using morphological, histochemical, immunohistochemical, and biochemical experiments. The preconditioned DPSC pellets exhibited significantly elevated levels of collagen II and glycosaminoglycans (GAGs) and reduced levels of the hypertrophic marker collagen X. No significant effect on GAGs production was observed in the preconditioned BMSC pellets, but collagen II and collagen X levels were elevated. While preconditioning did not modify the ALP specific activity in either cell type, it was notably lower in the DPSCs differentiated pellets compared to their BMSCs counterparts. These results could be interpreted as demonstrating the higher plasticity of DPSCs compared to BMSCs, suggesting the contribution of their unique molecular characteristics, including their negative Hox expression pattern, to promote a chondrogenic differentiation potential. Consequently, DPSCs could be considered compelling candidates for future cartilage cell therapy. [ABSTRACT FROM AUTHOR]

Background: Subarachnoid hemorrhage (SAH) is a severe stroke subtype that lacks effective treatment. Exosomes derived from human dental pulp stem cells (DPSCs) are a promising acellular therapeutic strategy for neurological diseases. However, the therapeutic effects of DPSC-derived exosomes (DPSC-Exos) on SAH remain unknown. In this study, we investigated the therapeutic effects and mechanisms of action of DPSC-Exos in SAH. Materials and methods: SAH was established using 120 male Sprague-Dawley rats. One hour after SAH induction, DPSC-Exos were administered via tail vein injection. To investigate the effect of DPSC-Exos, SAH grading, short-term and long-term neurobehavioral assessments, brain water content, western blot (WB), immunofluorescence staining, Nissl staining, and HE staining were performed. The role of miR-197-3p/FOXO3 in regulating pyroptosis was demonstrated through miRNA sequencing, bioinformatics analysis, and rescue experiments. The SAH model in vitro was established by stimulating BV2 cells with hemoglobin (Hb) and the underlying mechanism of DPSC-Exos was investigated through WB and Hoechst/PI staining. Results: The expressions of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) were increased after SAH. DPSC-Exos alleviated brain edema and neuroinflammation by inhibiting the expression of FOXO3 and reducing NLRP3 inflammasome activation, leading to improved neurobehavioral functions at 24 h after SAH. In vitro, the expression of the NLRP3 inflammasome components (NLRP3 and caspase1-p20), GSDMD-N, and IL-18 was inhibited in BV2 cells pretreated with DPSC-Exos. Importantly, DPSC-Exos overexpressing miR-197-3p had a more obvious protective effect than those from NC-transfected DPSCs, while those from DPSCs transfected with the miR-197-3p inhibitor had a weaker protective effect. Functional studies indicated that miR-197-3p bound to the 3ʹ-untranslated region of FOXO3, inhibiting its transcription. Furthermore, the overexpression of FOXO3 reversed the protective effects of miR-197-3p. Conclusions: DPSC-Exos inhibited activation of the NLRP3 inflammasome and related cytokine release via the miR-197-3p/FOXO3 pathway, alleviated neuroinflammation, and inhibited microglial pyroptosis. These findings suggest that using DPSC-Exos is a promising therapeutic strategy for SAH. [ABSTRACT FROM AUTHOR]

Rat dental pulp stem cells (DPSCs) can be used to elucidate mesenchymal stem cell (MSC) applications in regenerative medicine. However, information on rat DPSCs during long-term passage, which could lead to replicative senescence, is limited. In this study, we investigated the phenotypic changes in DPSCs after 3–26 passages (3P–26P). The results show that cell morphology and nuclear size increase proportionally with passage number. The phosphorylated histone H2A.X (γ-H2A.X) positive cells (indicating DNA damage) increased significantly earlier than the 4-Hydroxynonenal (4-HNE) stained cells (indicating an abundance of intracellular reactive oxygen species). Compared to the cells subjected to 3P and 5P, the cells subjected to 15P showed reduced proliferation despite being positive for Ki67. Furthermore, cell growth was completely arrested after 26P. The senescence markers, senescence-associated β-galactosidase (SA-β-gal) and p16, exhibited similar expression patterns that were not correlated with those of p21 and urokinase-type plasminogen activator receptor (uPAR). Nearly all cells expressed SA-β-gal and p16 after 26P, whereas only half expressed p21 and uPAR. These results will contribute to understanding the characteristics of DPSCs toward replicative senescence, which are applicable to elucidate mechanisms related to regenerative medicine and stem cell aging.

Objective To investigate the effect of human pulp stem cells （hDPSC) -derived exosomes （Exo） modified by circRNA （circRNA） signal-induced proliferation-associated protein-like 1（SIPA1L1） on the angiogenesis of human umbilical vein endothelial cells （HUVEC). Methods hDPSC was isolated and cultured from pulp tissue, the circSIPA1L1 overexpression plasmid was transfected into hDPSC, and Exo was isolated and identified. HUVEC was divided into control group, hDPSC Exo group and circSIPA1L1-hDPSC Exo group. After culture for 48 h, the vasculoformation ability was detected by Matrigel matrix gel vasculoformation test, the expression levels of vascular endothelial growth factor （VEGF), vascular endothelial growth factor receptor 2（VEGFR2） and placental growth factor （PGF） were determined by qRT-PCR and Western blot. Results The Exo was successfully isolated from the untransfected hDPSC and the hDPSC transfected with circSIPA1L1, and compared with the HDPSC-derived Exo, the relative expression of circSIPA1L1 in the hDPSC Exo transfected with circSIPA1L1 was significantly up-regulated （P < 0.05). Compared with hDPSC Exo group, the number of HUVEC tubular structures in circSIPA1L1-hDPSC Exo group was significantly increased （P < 0.05), and the mRNA and protein relative expressions of VEGF, VEGFR2 and PGF were also significantly up-regulated （P < 0.05）. Conclusions Modification of hDPSC-derived Exo by circRNA SIPA1L1 can promote angiogenesis, and the mechanism may be related to up-regulation of VEGF, VEGFR2 and PGF expression levels. [ABSTRACT FROM AUTHOR]

Objective: We aimed to identify the crucial genes involved in dental pulp stem cell (DPSC) senescence and evaluate the impact of melatonin on DPSC senescence. Methods: Western blotting, SA‐β‐Gal staining and ALP staining were used to evaluate the senescence and differentiation potential of DPSCs. The optimal concentration of melatonin was determined using the CCK‐8 assay. Differentially expressed genes (DEGs) involved in DPSC senescence were obtained via bioinformatics analysis, followed by RT–qPCR. Gain‐ and loss‐of‐function studies were conducted to explore the role of MMP3 in DPSC in vitro expansion and in response to melatonin. GSEA was employed to analyse MMP3‐related pathways in cellular senescence. Results: Treatment with 0.1 μM melatonin attenuated cellular senescence and differentiation potential suppression in DPSCs due to long‐term in vitro expansion. MMP3 was a crucial gene in senescence, as confirmed by bioinformatics analysis, RT–qPCR and Western blotting. Furthermore, gain‐ and loss‐of‐function studies revealed that MMP3 played a regulatory role in cellular senescence. Rescue assays showed that overexpression of MMP3 reversed the effect of melatonin on senescence. GSEA revealed that the MMP3‐dependent anti‐senescence effect of melatonin was associated with the IL6‐JAK‐STAT3, TNF‐α‐Signalling‐VIA‐NF‐κB, COMPLEMENT, NOTCH Signalling and PI3K‐AKT‐mTOR pathways. Conclusion: Melatonin attenuated DPSC senescence caused by long‐term expansion by inhibiting MMP3. [ABSTRACT FROM AUTHOR]

Background: Ageing is a key risk factor for cardiovascular disease and is linked to several alterations in cardiac structure and function, including left ventricular hypertrophy and increased cardiomyocyte volume, as well as a decline in the number of cardiomyocytes and ventricular dysfunction, emphasizing the pathological impacts of cardiomyocyte ageing. Dental pulp stem cells (DPSCs) are promising as a cellular therapeutic source due to their minimally invasive surgical approach and remarkable proliferative ability. Aim: This study is the first to investigate the outcomes of the systemic transplantation of DPSCs in a D-galactose (D-gal)-induced rat model of cardiac ageing. Methods. Thirty 9-week-old Sprague-Dawley male rats were randomly assigned into three groups: control, ageing (D-gal), and transplanted groups (D-gal + DPSCs). D-gal (300 mg/kg/day) was administered intraperitoneally daily for 8 weeks. The rats in the transplantation group were intravenously injected with DPSCs at a dose of 1 × 106 once every 2 weeks. Results: The transplanted cells migrated to the heart, differentiated into cardiomyocytes, improved cardiac function, upregulated Sirt1 expression, exerted antioxidative effects, modulated connexin-43 expression, attenuated cardiac histopathological alterations, and had anti-senescent and anti-apoptotic effects. Conclusion: Our results reveal the beneficial effects of DPSC transplantation in a cardiac ageing rat model, suggesting their potential as a viable cell therapy for ageing hearts. [ABSTRACT FROM AUTHOR]

Research on tooth regeneration using human-induced pluripotent stem cells (hiPSCs) is valuable for autologous dental regeneration. Acquiring mesenchymal and epithelial cells as a resource for dental regeneration is necessary because mesenchymal–epithelial interactions play an essential role in dental development. We reported the establishment of hiPSCs-derived dental epithelial-like cell (EPI-iPSCs), but hiPSCs-derived dental mesenchymal stem cells (MSCs) have not yet been reported. This study was conducted to establish hiPSCs-derived MSCs and to differentiate them into dental cells with EPI-iPSCs. Considering that dental MSCs are derived from the neural crest, hiPSCs were induced to differentiate into MSCs through neural crest formation to acquire the properties of dental MSCs. To differentiate hiPSCs into MSCs through neural crest formation, established hiPSCs were cultured and differentiated with PA6 stromal cells and differentiated hiPSCs formed neurospheres on ultralow-attachment plates. Neurospheres were differentiated into MSCs in serum-supplemented medium. Neural crest-mediated MSCs (NC-MSCs) continuously showed typical MSC morphology and expressed MSC markers. After 8 days of odontogenic induction, the expression levels of odontogenic/mineralization-related genes and dentin sialophosphoprotein (DSPP) proteins were increased in the NC-MSCs alone group in the absence of coculturing with dental epithelial cells. The NC-MSCs and EPI-iPSCs coculture groups showed high expression levels of amelogenesis/odontogenic/mineralization-related genes and DSPP proteins. Furthermore, the NC-MSCs and EPI-iPSCs coculture group yielded calcium deposits earlier than the NC-MSCs alone group. These results indicated that established NC-MSCs from hiPSCs have dental differentiation capacity with dental epithelial cells. In addition, it was confirmed that hiPSCs-derived dental stem cells could be a novel cell source for autologous dental regeneration. [ABSTRACT FROM AUTHOR]

BACKGROUND: Sema3A is a power secretory osteoprotective factor. However, studies about Sema3A-modified dental pulp stem cells (Sema3A-DPSCs) are rare. OBJECTIVE: To explore the osteogenic differentiation ability of Sema3A-DPSCs and their regulatory effect on the osteogenic differentiation of the pre-osteoblast cell line MC3T3-E1. METHODS: First, Sema3A-DPSCs were constructed using a lentivirus infection system carrying the Sema3A gene. Control lentivirus-treated DPSCs (Vector-DPSCs) were used as controls. Sema3A-DPSCs or Vector-DPSCs were co-cultured with proosteoblast line MC3T3-E1 at the ratio of 1 ∶ 1 and 1 ∶ 3 for 24 hours. Finally, the Sema3A-DPSCs, Vector-DPSCs and their co-cultured cells with MC3T3-E1 were cultured for osteogenic induction and differentiation. Osteogenic gene expression was detected by alkaline phosphatase staining, alizarin red staining and real-time quantitative RT-PCR to evaluate osteogenic differentiation ability. RESULTS AND CONCLUSION: (1) Sema3A mRNA and protein expression levels in Sema3A-DPSCs were significantly up-regulated. The level of secreted Sema3A in cell supernatant was up-regulated. (2) Compared with the Vector-DPSCs, mRNA expressions of osteogenic genes alkaline phosphatase, Runtrelated transcription factor 2, osteocalcin and Sp7 transcription factors in Sema3A-DPSCs were up-regulated; the activity of alkaline phosphatase was enhanced, and the formation of mineralized nodules increased. (3) There were no obvious differences in proliferation between Sema3A-DPSCs and VectorDPSCs. (4) Compared with MC3T3-E1/Vector-DPSCs co-culture system, the expression of MC3T3-E1 osteogenic genes was up-regulated, and the total alkaline phosphatase activity was enhanced and more mineralized nodules were formed in the MC3T3-E1/Sema3A-DPSCs co-culture system. (5) The results suggest that overexpression of Sema3A can enhance the osteogenic differentiation of DPSCs. Overexpression of Sema3A in DPSCs can promote osteogenic differentiation of MC3T3-E1 in the DPSCs/MC3T3-E1 co-culture system. [ABSTRACT FROM AUTHOR]

Cerebral ischemia-reperfusion injury (I/RI) is one of the principal pathogenic factors in the poor prognosis of ischemic stroke, for which current therapeutic options to enhance neurological recovery are notably insufficient. Dental pulp stem cell-derived extracellular vesicles (DPSC-EVs) have promising prospects in stroke treatment and the specific underlying mechanisms have yet to be fully elucidated. The present study observed that DPSC-EVs ameliorated the degree of cerebral edema and infarct volume by reducing the apoptosis of neurons. Furthermore, the miRNA sequencing and functional enrichment analysis identified that miR-877–3p as a key component in DPSC-EVs, contributing to neuroprotection and anti-apoptotic effects. Following target prediction and dual-luciferase assay indicated that miR-877–3p interacted with Bcl-2-associated transcription factor (Bclaf1) to play a function. The miR-877–3p inhibitor or Bclaf1 overexpression reversed the neuroprotective effects of DPSC-EVs. The findings reveal a novel therapeutic pathway where miR-877–3p, transferred via DPSC-EVs, confers neuroprotection against cerebral I/RI, highlighting its potential in promoting neuronal survival and recovery post-ischemia.

Background/purpose: Cell pyroptosis and gingival inflammation have been implicated in periodontitis progression. Our previous study revealed that AR-A014418, a pharmacological inhibitor of glycogen synthase kinase-3β (GSK-3β), can enhance the migratory and osteogenic differentiation abilities of rat dental pulp stem cells (rDPSCs). The present study aimed to explore the effect of AR on the inflammation of rDPSCs. Materials and methods: The primary rDPSCs were isolated and identified by flow cytometry, as well as Oil red O and Alizarin Red S staining. The rDPSCs were cultured and exposed to lipopolysaccharide (LPS) before treating them with different concentrations of AR-A014418. The cell viability was detected using the CCK-8 assay. The generation and secretion of pro-inflammatory cytokines (IL-18, TNF-α, L-1β, and IL-6) were examined by qPCR and ELISA, respectively. To investigate the activation of the NLRP3 inflammasome, the expression levels of pro-caspase 1, cleaved caspase 1, as well as NLRP3 were analyzed by western blotting and immunofluorescence, respectively. Results: In the rDPSCs, LPS prohibited cell viability and enhanced the generation and secretion of pro-inflammatory cytokines. LPS upregulated NLRP3 and cleaved caspase-1 protein levels and promoted ASC speck formation in the rDPSCs. AR-A014418 administration effectively blocked the LPS-induced inflammation of the rDPSCs in a dose-dependent way. Mechanistically, AR-A014418 significantly restrained the up-regulation of NLRP3 and cleaved caspase-1 in LPS-treated rDPSCs. Conclusion: Collectively, our findings suggest that AR-A014418 significantly mitigates LPS-induced inflammation of rDPSCs by blocking the activation of the NLRP3 inflammasome.

Background Ageing is a key risk factor for cardiovascular disease and is linked to several alterations in cardiac structure and function, including left ventricular hypertrophy and increased cardiomyocyte volume, as well as a decline in the number of cardiomyocytes and ventricular dysfunction, emphasizing the pathological impacts of cardiomyocyte ageing. Dental pulp stem cells (DPSCs) are promising as a cellular therapeutic source due to their minimally invasive surgical approach and remarkable proliferative ability. Aim This study is the first to investigate the outcomes of the systemic transplantation of DPSCs in a D-galactose (D-gal)-induced rat model of cardiac ageing. Methods. Thirty 9-week-old Sprague-Dawley male rats were randomly assigned into three groups: control, ageing (D-gal), and transplanted groups (D-gal + DPSCs). D-gal (300 mg/kg/day) was administered intraperitoneally daily for 8 weeks. The rats in the transplantation group were intravenously injected with DPSCs at a dose of 1 × 106 once every 2 weeks. Results The transplanted cells migrated to the heart, differentiated into cardiomyocytes, improved cardiac function, upregulated Sirt1 expression, exerted antioxidative effects, modulated connexin-43 expression, attenuated cardiac histopathological alterations, and had anti-senescent and anti-apoptotic effects. Conclusion Our results reveal the beneficial effects of DPSC transplantation in a cardiac ageing rat model, suggesting their potential as a viable cell therapy for ageing hearts.

Aim: To investigate the effect of IWP‐2, Wnt inhibitor, on human dental pulp stem cells (hDPSCs) responses. Methodology: hDPSCs were isolated from human dental pulp tissues. Cells were treated with 25 μM IWP‐2 for 24 h, and subsequently, the gene expression profile was examined using high‐throughput RNA sequencing. The mRNA expression was analysed using qPCR. The effect of IWP‐2 was investigated in both normal and LPS‐induced hDPSCs (inflamed hDPSCs). CD4+ T cells and CD14+ monocyte‐derived macrophages were cultured with conditioned media of IWP‐2 treated hDPSCs to observe the immunosuppressive property. Results: RNA sequencing indicated that IWP‐2 significantly downregulated several KEGG pathways, including cytokine‐cytokine receptor interaction, IL‐17 signalling pathway, and TNF signalling pathway. In both normal and inflamed conditions, IWP‐2 markedly upregulated TGFB1 mRNA expression while the mRNA expression of pro‐inflammatory cytokines, TNFA, IL1B, IFNG, and IL6, was inhibited. In the inhibition experiment, the pretreatment with p38, MAPK, or PI3K inhibitors abolished the effects of IWP‐2 in LPS‐induced inflammation. In terms of immune cells, IWP‐2‐treated‐inflamed hDPSCs conditioned media attenuated T cell proliferation and regulated regulatory T cell differentiation. In addition, the migratory property of macrophage was decreased after being exposed to IWP‐2‐treated inflamed hDPSCs conditioned media. Conclusion: IWP‐2 suppressed inflammatory cytokine expression in both normal and inflamed hDPSCs. Moreover, hDPSCs exerted the immunosuppressive property after IWP‐2 treatment. These results suggest the role of Wnt in inflammatory responses and immunomodulation in dental pulp tissues. [ABSTRACT FROM AUTHOR]

Background/purpose: Human dental pulp stem cells (hDPSCs) are an emerging source of mesenchymal stem cells (MSCs) for bone tissue regeneration and engineering. In bone regeneration using transplanted MSCs, the extracellular environment or co-injected drugs can affect their success or failure. In this study, we investigated the effects and signaling mechanisms of lidocaine on osteogenic differentiation of hDPSCs after inducing inflammatory conditions with lipopolysaccharide (LPS) and tumor necrosis factor-alpha (TNF-α). Materials and methods: To investigate the effect of lidocaine on the osteogenic differentiation of LPS/TNF-α-treated hDPSCs, alkaline phosphatase (ALP) and Alizarin red S (ARS) staining were conducted. The expression of osteogenesis-related genes was assessed using quantitative real-time polymerase chain reaction and western blotting. The expression of mitogen-activated protein kinases was analyzed to evaluate the effect of lidocaine on osteogenic differentiation of LPS/TNF-α-treated hDPSCs. Results: Various concentrations of lidocaine (0.05, 0.2, and 1 mM) further decreased ALP and ARS staining of LPS/TNF-α-treated hDPSCs. Similarly, the mRNA and protein expression of osteogenesis-related genes was suppressed via lidocaine treatment in LPS/TNF-α-treated hDPSCs. Lidocaine treatment downregulated the protein expression of p-ERK and p-JNK in LPS/TNF-α-treated hDPSCs. Conclusion: Lidocaine intensified the inhibition of osteogenic differentiation on inflammation-induced hDPSCs by inhibiting the ERK and JNK signaling pathways. This in vitro study suggested that lidocaine may have an inhibitory effect on bone regeneration.

BACKGROUND: Dental pulp stem cells have excellent characteristics of multi-directional differentiation. Among them, osteogenic differentiation potential has a good prospect in bone tissue engineering and regeneration therapy, while inflammatory factors are strongly associated with it. OBJECTIVE: To review the research progress on the bidirectional effects of inflammatory factors and osteogenic differentiation of dental pulp stem cells during bone regeneration. METHODS: In the PubMed, CNKI, and Wanfang databases, the search terms were “dental pulp stem cells, inflammation, osteogenic differentiation, bone regeneration”in Chinese and “dental pulp stem cell*, dpsc*, inflam*, osteogenic differentiation, ossification, osteogenesis, bone formation” in English. Relevant articles published from 2012 to 2022 were retrieved, and 76 articles were finally included for review. RESULTS AND CONCLUSION: (1) As dental mesenchymal stem cells, dental pulp stem cells have well-known multi-directional differentiation characteristics, among which the osteogenic differentiation potential is a research hotspot of bone tissue engineering. (2) Inflammatory factors are strongly associated with bone regeneration mediated by dental pulp stem cells whether they are the microenvironment where cells live, the adjustable immune inflammatory response, or the inflammatory disease with bone tissue defect that needs regeneration. In recent years, there has been no lack of related research. (3) This article selected relevant articles in the past decade, and found that even inflammatory dental pulp stem cells from inflamed pulp tissue still retain certain osteogenic differentiation potential. Further analysis of the bidirectional relationship between inflammatory factors and dental pulp stem cells during bone regeneration found that the effect of inflammatory factors on the osteogenic differentiation of dental pulp stem cells and the immune regulation of dental pulp stem cells on inflammation have both promoting and inhibiting effects. This is mainly related to the type, concentration and time of inflammatory mediators, but the specific mechanisms are not comprehensive. (4) Recently, scholars’ researches focus on exploring the promoting factors that mediate the osteogenic differentiation of dental pulp stem cells in an inflammatory environment, especially the exogenous factors in addition to inflammatory mediators, such as drugs. (5) Some animal experiments and a few clinical trials have verified the therapeutic effect of dental pulp stem cells on inflammatory bone defect diseases especially periodontitis and arthritis by using the anti-inflammatory and osteogenic properties of dental pulp stem cells. (6) At present, there is still a big gap in the research on the mechanism of the bidirectional interaction between inflammatory factors and osteogenic differentiation of dental pulp stem cells during bone regeneration, which needs further investigation and verification in the future. [ABSTRACT FROM AUTHOR]

Dental pulp stem cells (DPSCs) are a type of mesenchymal stem cells that can differentiate into odontoblast-like cells and protect the pulp. The differentiation of DPSCs can be influenced by biomaterials or growth factors that activate different signaling pathways in vitro or in vivo. In this review, we summarized six major pathways involved in the odontogenic differentiation of DPSCs, Wnt signaling pathways, Smad signaling pathways, MAPK signaling pathways, NF-kB signaling pathways, PI3K/AKT/mTOR signaling pathways, and Notch signaling pathways. Various factors can influence the odontogenic differentiation of DPSCs through one or more signaling pathways. By understanding the interactions between these signaling pathways, we can expand our knowledge of the mechanisms underlying the regeneration of the pulp-dentin complex. [ABSTRACT FROM AUTHOR]

Background: Conditioned media (CM) derived from mesenchymal stem cells (MSC) is known to induce hair regrowth in androgenic alopecia. Objectives: The objectives of the study were to assess the efficacy and safety of one type of MSC‐CM, the CM derived from dental pulp stem cells obtained from human exfoliated deciduous teeth (SHED‐CM) and to compare the efficacy of SHED‐CM with and without dihydrotestosterone synthesis inhibitor (DHT‐inhibitor). Methods: Eighty‐eight male androgenic alopecia subjects with Hamilton‐Norwood Classification (H‐N C) I–VII were evaluated by trichoscopy to explore which trichoscopic factors statistically correlated with H‐N C. After being screened, 33 subjects received six SHED‐CM treatments at 1‐month intervals. Clinical severity was assessed through global and trichoscopic images from baseline to 9th month. Results: SHED‐CM was effective for 75% of subjects regardless of disease severity, concomitant DHT‐inhibitor use, and age. Adverse effects including pain and small hemorrhages were transient and mild. We also found that clinical hair status evaluated by absolute values of three quantitative trichoscopic factors (maximum hair diameter, vellus hair rate, and multi‐hair follicular unit rate) showed a good correlation with H‐N C stages, and what is more—a scoring system of these three factors can be a possible predictor of SHED‐CM efficacy. Conclusions: We have shown that SHED‐CM provides global and trichoscopic image improvement for androgenic alopecia, regardless of concomitant DHT‐inhibitor use. [ABSTRACT FROM AUTHOR]

Objectives: In this study, it was aimed to investigate the possible effects of oral chewable probiotic tablets (PTs) produced to directly support the oral flora on the proliferation of human dental pulp stem cells (DPSCs) and human gingival fibroblast cells (HGFCs). Materials and Methods: For analysis in this study, "Motiflor AS," a PT that dissolves in the mouth, containing 13.5mg Lactobacillus helveticus Rosell-52, L. rhamnosus Rosell-11, L. halivarus HA-118, and Bifidobacterium longum Rosell-175 was used. Cell survival and proliferation were analyzed by methyl-thiazole-diphenyl-tetrazolium (MTT) test and real-time cell analysis method (xCELLigence RTCA-DP) after 24-, 48-, and 72-h incubation periods. Results: According to the data obtained with RTCA-DP software, there was a significant increase in the proliferation of human dental pulp stem cells (HDPSCs) and HGFCs in the 72-h incubation after PT application compared to the 24-h and 48-h incubations (P < 0.0001). After the MTT test, for HDPSCs, the cell proliferation rate was 62.8% and 85.6% in 24- and 48-h incubation, respectively, while HDPSCs cell proliferation rate in 72-h incubation was 135.2% (P < 0.0001). For HGFCs, the cell proliferation rate was 73% and 120.4% in 24- and 48-h incubation, respectively, while HDPSCs cell proliferation rate in 72-h incubation was 139.8% (P < 0.0001). When the results of the two tests applied were evaluated together, the results showed compatibility. Conclusions: Based on the results, it has been concluded that PT will be useful for maintaining oral health and for dental and gingival patients who will/have undergone dental treatment. It should be keep in mind that protecting our oral and dental health is very important in terms of protecting our general health. [ABSTRACT FROM AUTHOR]

BACKGROUND: Revascularization plays an important role in tissue engineering. Dental pulp chamber has its unique anatomy structure, which is surrounded by hard dentin. The blood supply of dental pulp only comes from the narrow apical opening, which makes revascularization more difficult in dental pulp regeneration. OBJECTIVE: To conclude the research advances focusing on the designs of signal molecules, scaffolds and cells in promoting vascularization in dental pulp regeneration. METHODS: We searched the articles on PubMed and CNKI databases with the keywords of “dental pulp regeneration, revascularization, growth factor, material, scaffold, dental pulp stem cell, prevascularization, coculture” in Chinese and English, respectively. Finally, 69 articles met the criteria for review. RESULTS AND CONCLUSION: (1) Vascular endothelial growth factor, platelet-derived growth factor and fibroblast growth factor are the most studied proangiogenic factors, which promote angiogenesis by promoting the proliferation and migration of endothelial and perivascular cells, the recruitment of stem cells and their differentiation to endothelial cells. (2) To maintain the appropriate concentration of pro-angiogenic factors in the microenvironment, the addition of heparin, laponite and cellulose nanocrystals to the scaffolds can solubilize, encapsulate, adsorb, and covalently cross-link signal molecules, which mimic in vivo storage and release mechanisms. Natural materials including decellularized dental pulp matrix, platelet-rich plasma, and platelet-rich fibrin are rich in bioactive molecules and their structures can protect and control the release of these signal molecules. (3) Different cell sub-populations of dental pulp stem cells have different angiogenic potentials. Moreover, stem cells can be transfected by biological and chemical vectors to improve the cells’ ability to secrete signal molecules and to promote the transformation of stem cells to endothelial cells. Therefore, the angiogenic capacity of transfected cells is significantly enhanced. Co-implantation of stem cells and endothelial cells can also enhance angiogenic capacity. (4) Prevascularization may be the most powerful means to promote rapid vascularization of dental pulp regeneration. In vitro co-culture of endothelial cells and stem cells with a certain ratio can form microtissues containing vascular structures. Microvascular fragments can be obtained by enzymatic digestion of adipose tissue, in which tubular structures and functional cells can promote the development of new blood vessels and anastomosis with host vessels. (5) Current clinical studies on revascularization of dental pulp are limited to dental pulprevascularization, where signal molecules are added while stimulating the formation of blood clots in the root canal and the effects are observed. While other research directions mainly involve in vivo trials in small and large animals, and further clinical trials are still needed to observe regenerative effects, mitigate side effects, and optimize the preparation process. [ABSTRACT FROM AUTHOR]

Cell pyroptosis and gingival inflammation have been implicated in periodontitis progression. Our previous study revealed that AR-A014418, a pharmacological inhibitor of glycogen synthase kinase-3β (GSK-3β), can enhance the migratory and osteogenic differentiation abilities of rat dental pulp stem cells (rDPSCs). The present study aimed to explore the effect of AR on the inflammation of rDPSCs. The primary rDPSCs were isolated and identified by flow cytometry, as well as Oil red O and Alizarin Red S staining. The rDPSCs were cultured and exposed to lipopolysaccharide (LPS) before treating them with different concentrations of AR-A014418. The cell viability was detected using the CCK-8 assay. The generation and secretion of pro-inflammatory cytokines (IL-18, TNF-α, L-1β, and IL-6) were examined by qPCR and ELISA, respectively. To investigate the activation of the NLRP3 inflammasome, the expression levels of pro-caspase 1, cleaved caspase 1, as well as NLRP3 were analyzed by western blotting and immunofluorescence, respectively. In the rDPSCs, LPS prohibited cell viability and enhanced the generation and secretion of pro-inflammatory cytokines. LPS upregulated NLRP3 and cleaved caspase-1 protein levels and promoted ASC speck formation in the rDPSCs. AR-A014418 administration effectively blocked the LPS-induced inflammation of the rDPSCs in a dose-dependent way. Mechanistically, AR-A014418 significantly restrained the up-regulation of NLRP3 and cleaved caspase-1 in LPS-treated rDPSCs. Collectively, our findings suggest that AR-A014418 significantly mitigates LPS-induced inflammation of rDPSCs by blocking the activation of the NLRP3 inflammasome. [ABSTRACT FROM AUTHOR]

Dental pulp stem cells (DPSCs) are considered a valuable cell source for regenerative medicine because of their high proliferative potential, multipotency, and availability. We established a new cryopreservation method (NCM) for collecting DPSCs, in which the tissue itself is cryopreserved and DPSCs are collected after thawing. We improved the NCM and developed a new method for collecting and preserving DPSCs more efficiently. Dental pulp tissue was collected from an extracted tooth, divided into two pieces, sandwiched from above and below using cell culture inserts, and cultured. As a result, the cells in the pulp tissue migrated vertically over time and localized near the upper and lower membranes over 2–3 days. With regard to the underlying molecular mechanism, SDF1 was predominantly involved in cell migration. This improved method is valuable and enables the more efficient collection and reliable preservation of DPSCs. It has the potential to procure a large number of DPSCs stably. [ABSTRACT FROM AUTHOR]

Copyright of Ege Journal of Medicine is the property of Ege University, Faculty of Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

BACKGROUND: How to effectively repair and reconstruct critical bone defects has become a difficult problem in clinical medicine. Dental pulp stem cells have certain advantages as transgenic target cells to repair damaged tissues or organs. OBJECTIVE: To investigate the effect of hypoxia-inducible factor-1α on osteogenesis and angiogenesis of dental pulp stem cells in vitro. METHODS: The pulp tissues of healthy and intact bicuspid teeth extracted by orthodontic treatment in patients aged 12 to 18 years were isolated to culture dental pulp stem cells. The hypoxia-inducible factor-1α gene was transfected into dental pulp stem cells in the experimental group. The control group did not do any treatment. The expression of the hypoxia-inducible factor 1α gene was detected by qPCR and the expression of osteogenic and angiogenic factors was detected by western blot assay. RESULTS AND CONCLUSION: (1) Dental pulp stem cells were short spindle-shaped fibroblast-like cells, and the cells grew in colonies. (2) Hypoxia-inducible factor-1α gene was stably transfected into dental pulp stem cells. The expression of the hypoxia-inducible factor-1α gene increased gradually with the extension of time, reached a peak at 14 days, and significantly decreased at 21 days. (3) Compared with the control group, the expression level of angiogenesis-related factors in the experimental group was higher. The protein expression levels of vascular endothelial growth factor, stromal cell-derived factor 1, angiopoietin 2, and placental growth factor were significantly increased at 1 day after gene transfection and reached a peak at 4 days. After that, the expression of angiogenesis-associated protein in the experimental group decreased slightly, but was significantly higher than that in the control group. (4) Compared with the control group, the expression level of osteogenic-related factors in the experimental group was higher. The protein expression levels of human bone morphogenetic protein 2, osteocalcin, and Runt-related transcription factor 2 were significantly increased at 1 day after gene transfection, with the highest protein expression at 4 days. (5) It is concluded that hypoxia-inducible factor-1α can promote the osteogenesis and the expression of angiogenic factors in dental pulp stem cells. [ABSTRACT FROM AUTHOR]

Amyotrophic lateral sclerosis (ALS) is a fatal and incurable paralytic disorder caused by the progressive death of upper and lower motoneurons. Although numerous strategies have been developed to slow disease progression and improve life quality, to date only a few therapeutic treatments are available with still unsatisfactory therapeutic benefits. The secretome of dental pulp stem cells (DPSCs) contains numerous neurotrophic factors that could promote motoneuron survival. Accordingly, DPSCs confer neuroprotective benefits to the SOD1G93A mouse model of ALS. However, the mode of action of DPSC secretome on motoneurons remains largely unknown. Here, we used conditioned medium of human DPSCs (DPSCs-CM) and assessed its effect on survival, axonal length, and electrical activity of cultured wildtype and SOD1G93A motoneurons. To further understand the role of individual factors secreted by DPSCs and to circumvent the secretome variability bias, we focused on GDF15 and HB-EGF whose neuroprotective properties remain elusive in the ALS pathogenic context. DPSCs-CM rescues motoneurons from trophic factor deprivation-induced death, promotes axon outgrowth of wildtype but not SOD1G93A mutant motoneurons, and has no impact on the spontaneous electrical activity of wildtype or mutant motoneurons. Both GDF15 and HB-EGF protect SOD1G93A motoneurons against nitric oxide-induced death, but not against death induced by trophic factor deprivation. GDF15 and HB-EGF receptors were found to be expressed in the spinal cord, with a two-fold increase in expression for the GDF15 low-affinity receptor in SOD1G93A mice. Therefore, the secretome of DPSCs appears as a new potential therapeutic candidate for ALS. [ABSTRACT FROM AUTHOR]

Dental pulp stem cells (DPSCs) are a type of mesenchymal stem cells that can differentiate into odontoblast-like cells and protect the pulp. The differentiation of DPSCs can be influenced by biomaterials or growth factors that activate different signaling pathways in vitro or in vivo. In this review, we summarized six major pathways involved in the odontogenic differentiation of DPSCs, Wnt signaling pathways, Smad signaling pathways, MAPK signaling pathways, NF-kB signaling pathways, PI3K/AKT/mTOR signaling pathways, and Notch signaling pathways. Various factors can influence the odontogenic differentiation of DPSCs through one or more signaling pathways. By understanding the interactions between these signaling pathways, we can expand our knowledge of the mechanisms underlying the regeneration of the pulp–dentin complex.

BACKGROUND: Resveratrol is a natural non-flavonoid polyphenol compound. Studies have shown that resveratrol has antioxidant, anti-inflammatory, anti-aging and anti-tumor activities, and promotes the osteogenic differentiation of bone marrow mesenchymal stem cells. However, whether resveratrol can promote the osteogenic differentiation of stem cells from human exfoliated deciduous teeth is even not expounded. OBJECTIVE: To investigate the effects of resveratrol on the proliferation and osteogenic differentiation of stem cells from human exfoliated deciduous teeth. METHODS: Human stem cells from human exfoliated deciduous teeth were isolated and cultured by tissue block method. Osteogenic induced stem cells from human exfoliated deciduous teeth were treated with different concentrations of resveratrol. Cell activity was detected by CCK-8 assay. The number of mineralized nodular cells was detected by alizarin red staining. mRNA expression of osteogenic genes was detected by RT-qPCR and protein expression of Runtrelated transcription factor 2 was detected by western blot assay. RESULTS AND CONCLUSION: (1) CCK-8 assay results showed that after 3 days of treatment, resveratrol at concentrations of (20, 40, 80, and 100 μmol/L) significantly inhibited the proliferation of stem cells from human exfoliated deciduous teeth. Therefore, 1, 5, and 10 μmol/L resveratrol could be used to treat stem cells from human exfoliated deciduous teeth in subsequent experiments to detect their effects on osteogenic differentiation. (2) With 10 μmol/L resveratrol treatment, the number of osteogenic nodules of stem cells from human exfoliated deciduous teeth increased significantly compared with the control group. (3) Alkaline phosphatase, Runt-related transcription factor 2, and osteocalcin mRNA expression levels were increased after 10 μmol/L resveratrol treatment compared with the control group. (4) Runt-related transcription factor 2 protein expression levels of resveratrol treatment groups increased in a concentration-dependent manner compared with the control group. (5) The results indicate that appropriate concentration of resveratrol promotes the proliferation and osteogenic differentiation of stem cells from human exfoliated deciduous teeth. [ABSTRACT FROM AUTHOR]

BACKGROUND: The specific regulatory mechanism underlying osteogenic differentiation of human dental pulp stem cells is still unclear. Therefore, studying the specific mechanism is of great significance for the application of stem cells in tissue engineering. OBJECTIVE: To investigate the role of lentivirus mediated silencing of Smad2 gene in the osteogenic differentiation of human dental pulp stem cells. METHODS: The third generation of human dental pulp stem cells were cultured and divided into blank control group, negative control group, Smad2-shRNA group, and Smad2-shRNA+transforming growth factor-β3 (TGF-β3) group. The mRNA expression of osteogenic related genes Runx2, osteocalcin, Smad2, Smad3, and Smad4 in human dental pulp stem cells was measured by qRT-PCR. The protein expression levels of Runx2, osteocalcin, Smad2/Smad3, p-Smad2/ Smad3, and Smad4 in human dental pulp stem cells were measured by western blot assay. Alizarin red staining was performed in each group on the 7th and 14th days of culture. RESULTS AND CONCLUSION: (1) The mRNA expressions of Runx2, osteocalcin, Smad2, Smad3, and Smad4 in the Smad2-shRNA group were significantly lower than those in the negative control group and blank control group (P < 0.05). The mRNA expressions of osteocalcin, Smad2, Smad3, and Smad4 were significantly increased in the Smad2-shRNA+TGF-β3 group compared with the Smad2-shRNA group (P < 0.05). (2) The protein expressions of Runx2, osteocalcin, and Smad2/Smad3 in the Smad2-shRNA group were significantly lower than those in the negative control group and blank control group (P < 0.05). The protein expressions of RUNX2, osteocalcin, Smad2/Smad3, and Smad4 were significantly increased in the Smad2-shRNA+TGF-β3 group compared with the Smad2- shRNA group (P < 0.05). (3) Culture with conventional medium and transfection with lentiviral vector could not induce the osteogenic differentiation of human dental pulp stem cells, while TGF-β3 could positively regulate the osteogenic differentiation of human dental pulp stem cells. (4) All these findings indicate that theTGF-β/Smad2 signal transduction pathway plays an important role in the osteogenic differentiation of human pulp stem cells. [ABSTRACT FROM AUTHOR]

Objective: Human dental pulp stem cells (hDPSCs) constitute a promising source of stem cells in tissue engineering. However, the molecular mechanism of differentiation in hDPSCs remains largely unclear. MicroRNAs (miRNAs) play crucial roles in lineage‐specific differentiation of stem cells. The present study investigated the function of miRNA‐342‐5p in the odonto/osteogenic differentiation of hDPSCs. Methods: The miRNA array profiling and quantitative real‐time reverse transcriptase‐polymerase chain reaction (qRT‐PCR) revealed the expression of miR‐342‐5p during odonto/osteogenic differentiation of hDPSCs. hDPSCs were treated with miR‐342‐5p mimic and inhibitor to investigate the regulatory roles of miR‐342‐5p in the differentiation of hDPSCs. Moreover, miR‐342‐5p inhibitor and small interference RNA (siRNA) targeting Wnt7b were applied to explore the regulatory mechanism of miR‐342‐5p. Results: Downregulated miR‐342‐5p was observed during odonto/osteogenic differentiation of hDPSCs. The overexpression of miR‐342‐5p inhibited the odonto/osteogenic potential of DPSCs, as indicated by low levels of alkaline phosphatase activity, calcium deposition formation, and odonto/osteogenic differentiation markers, whereas silencing of miR‐342‐5p exhibited the opposite effect. When co‐treated with siRNA targeting Wnt7b and miR‐342‐5p inhibitor in hDPSCs, the odonto/osteogenic potential and activation of Wnt7b/β‐catenin pathway were attenuated. Conclusions: This study showed that miR‐342‐5p inhibits the odonto/osteogenic differentiation of hDPSCs by interfering with Wnt/β‐catenin signaling via targeting Wnt7b. [ABSTRACT FROM AUTHOR]

Human dental pulp stem cells (hDPSCs) are an emerging source of mesenchymal stem cells (MSCs) for bone tissue regeneration and engineering. In bone regeneration using transplanted MSCs, the extracellular environment or co-injected drugs can affect their success or failure. In this study, we investigated the effects and signaling mechanisms of lidocaine on osteogenic differentiation of hDPSCs after inducing inflammatory conditions with lipopolysaccharide (LPS) and tumor necrosis factor-alpha (TNF-α). To investigate the effect of lidocaine on the osteogenic differentiation of LPS/TNF-α-treated hDPSCs, alkaline phosphatase (ALP) and Alizarin red S (ARS) staining were conducted. The expression of osteogenesis-related genes was assessed using quantitative real-time polymerase chain reaction and western blotting. The expression of mitogen-activated protein kinases was analyzed to evaluate the effect of lidocaine on osteogenic differentiation of LPS/TNF-α-treated hDPSCs. Various concentrations of lidocaine (0.05, 0.2, and 1 mM) further decreased ALP and ARS staining of LPS/TNF-α-treated hDPSCs. Similarly, the mRNA and protein expression of osteogenesis-related genes was suppressed via lidocaine treatment in LPS/TNF-α-treated hDPSCs. Lidocaine treatment downregulated the protein expression of p-ERK and p-JNK in LPS/TNF-α-treated hDPSCs. Lidocaine intensified the inhibition of osteogenic differentiation on inflammation-induced hDPSCs by inhibiting the ERK and JNK signaling pathways. This in vitro study suggested that lidocaine may have an inhibitory effect on bone regeneration. [ABSTRACT FROM AUTHOR]

Porphyromonas gingivalis is associated with endodontic pulpitis, causing damage to the dental pulp, leading to severe pain and a decline in quality of life. Regenerative pulp treatments using dental pulp stem cells (DPSCs) can be hindered by interactions between DPSCs and the infecting bacteria. The protein WNT family member 4 (Wnt4) plays a critical role in the differentiation of DPSCs and the regeneration of odontogenic tissue. However, the specific influence of P. gingivalis on Wnt4 remains unclear. In this study, we employed a computational approach to investigate the underlying mechanisms through which P. gingivalis-produced metabolites inhibit the Wnt4 protein, thereby diminishing the regenerative potential and therapeutic efficacy of odontogenic tissue. Among the metabolites examined, C29H46N7O18P3S−4 exhibited the strongest inhibitory effect on the Wnt4 protein, as evidenced by the lowest binding energy score of −6782 kcal/mol. Molecular dynamic simulation trajectories revealed that the binding of C29H46N7O18P3S−4 significantly altered the structural dynamics and stability of the Wnt4 protein. These alterations in protein trajectories may have implications for the molecular function of Wnt4 and its associated pathways. Overall, our findings shed light on the inhibitory impact of P. gingivalis-produced metabolites on the Wnt4 protein. Further in vitro, in vivo, and clinical studies are necessary to validate and expand upon our findings. [ABSTRACT FROM AUTHOR]

Maintaining the viability and avoiding necrosis of dental pulp are crucial to preserving the structural integrity and functioning of teeth. In recent years, cell-based regenerative endodontics has emerged as a promising approach to achieve this goal and has gained increasing attention in scientific research; however, in the confined space of the root canal system, hypoxic conditions can be both beneficial and detrimental, as they may promote angiogenesis in the graft to some extent but also lead to tissue necrosis if prolonged. Dental pulp stem cells (DPSCs) have been verified as multipotent cells that can promote angiogenesis and are therefore ideal candidates for realising real dental pulp regeneration within root canals. Thus, we focus on the underlying mechanisms of DPSCs to promote angiogenesis and summarise some preclinical studies and clinical trials involving transplanting of DPSCs to achieve real dental pulp regeneration, in the hope that this intractable source of perplexity in regenerative endodontics may be resolved sooner. [ABSTRACT FROM AUTHOR]

The odontogenic differentiation of dental pulp stem cells (DPSCs), which is vital for tooth regeneration, was regulated by various functional molecules. In recent years, a growing body of research has shown that miRNAs play a crucial role in the odontogenic differentiation of human dental pulp stem cells (hDPSCs). However, the mechanisms by which miRNAs regulated odontogenic differentiation of hDPSCs remained unclear, and the application of miRNAs in reparative dentin formation in vivo was also rare. In this study, we first discovered that miR-3074-3p had an inhibitory effect on odontogenic differentiation of hDPSCs and antagomiR-3074-3p-conjugated PEI-AuNPs effectively promoted odontogenic differentiation of hDPSCs in vitro. AntagomiR-3074-3p-conjugated PEI-AuNPs was further applied to the rat pulp-capping model and showed the increased formation of restorative dentin. In addition, the results of lentivirus transfection in vitro suggested that FKBP9 acted as the key target of miR-3074-3p in regulating the odontogenic differentiation of hDPSCs. These findings might provide a new strategy and candidate target for dentin restoration and tooth regeneration. [ABSTRACT FROM AUTHOR]

Background/purpose: Various studies have used stem cells in the field of bone tissue engineering to repair bone defects. Dental pulp stem cells (DPSCs) have multipotent properties and can be acquired in a noninvasive manner; therefore, they are frequently used in experiments in regenerative medicine. The objective of this study was to investigate the odontogenic/osteogenic differentiation of human DPSCs (hDPSCs) using propofol, a widely used intravenous anesthetic agent. Materials and methods: Alkaline phosphatase (ALP) staining was used to investigate the effects of various concentrations of propofol (5, 20, 50 and 100 μM) on the osteogenic differentiation of hDPSCs. Real-time qPCR and Western blot analysis were used to detect the effect of propofol on the expression of odontogenic/osteogenic genes, such as DMP1, RUNX2, OCN, and BMP2. Odontogenic/osteogenic differentiation of hDPSCs was estimated at days 7 and 14. Results: ALP staining of hDPSCs was significantly decreased by propofol treatment. The mRNA expression of DMP1, RUNX2, OCN, and BMP2 decreased after propofol treatment for 14 days. The protein expression of DMP1 and BMP2 was decreased by propofol at days 7 and 14, and that of RUNX2 was decreased by propofol at day 14 only. Conclusion: Propofol attenuated odontogenic/osteogenic differentiation of hDPSCs in vitro. This result suggests that propofol, which is widely used for dental sedation, may inhibit the odontogenic/osteogenic differentiation of hDPSCs.

Abstract Background Bone has important functions in the body. Several researchers have reported that the polysaccharides and lipopolysaccharide derived from microbes can promote osteogenic differentiation of stem cells. Enterococcus faecium, a lactic acid bacterium (LAB), produces several bioactive metabolites and has been widely applied in the food and nutraceutical industries. The exopolysaccharide (EPS) from LAB has also been extensively examined for its postbiotic effects and for its in vivo and in vitro functionalities. However, studies on promoting bone differentiation using polysaccharides from LAB are lacking. Therefore, the purpose of this study was to investigate the effect of E. faecium L15 extract and EPS on osteogenic differentiation of human dental pulp stem cells (hDPSCs) and to identify the underlying mechanisms. Methods hDPSCs were obtained from dental pulp tissue, and L15 extract and EPS were isolated from L15. Gene and protein expression of the osteogenic differentiation markers were analyzed with qPCR and western blotting and the possible signaling pathways were also investigated using western blotting. Osteogenic differentiation potential was examined by alkaline phosphatase (ALP) staining and alizarin red s (ARS) staining. In addition, osteogenic differentiation potential of L15 EPS was explored in ex vivo culture of neonate murine calvaria. Results The calcium deposition and ALP activity were enhanced by addition of L15 extract or EPS. The expression levels of RUNX2, ALP, and COL1A1 mRNA and the protein expression levels of RUNX2, ALP, and BMP4 were increased in hDPSCs treated with the L15 extract or EPS. The L15 EPS treatment enhanced phosphorylation of the p38 mitogen-activated protein kinase (MAPK). The L15 EPS-induced increases in RUNX2, ALP, and BMP4 expression were suppressed by the p38 MAPK inhibitor SB203580. The promoting effect of L15 EPS on osteogenic differentiation was not only seen in hDPSCs, but also in osteoblast precursors. ALP activity and the expression of RUNX2, ALP, and COL1A1 increased in the L15 EPS-treated osteoblast precursors. In addition, L15 EPS increased bone thickness of neonate murine calvaria in ex vivo culture. Conclusions The stimulatory effect of L15 extract and EPS on osteogenic differentiation occurred through the p38 MAPK pathway, and L15 EPS enhanced new bone formation in neonate murine calvaria. These data suggest that L15 EPS has therapeutic potential applicable to bone regeneration.

OBJECTIVE: To summarize research progress on forming dental pulp-like tissue from dental pulp stem cells via a co-culture system, as well as provide ideas for stem cell therapy for dental pulp tissue regeneration. METHODS: The first author searched PubMed, CNKI, and Wanfang databases for relevant literature published before May 2022. The English search terms were “dental pulp regeneration, dental pulp stem cell, co-culture, tissue engineering, signaling pathway”, and the Chinese search terms were “dental pulp regeneration, dental pulp stem cells, co-culture, tissue engineering, scaffold, signaling pathway”. Totally 67 articles meeting the criteria were summarized and described. RESULTS AND CONCLUSION: (1) Different combinations of cells are used in co-culture systems to form the main body of the cell and the combined body. (2) Direct co-culture and indirect co-culture are the two types of combination modes at present. Through the intervention of biological scaffold materials and a change in culture conditions, the co-culture outcome can be divided into two-dimensional regenerated tissues and three-dimensional regenerated tissues. (3) Direct co-culture creates an extracellular matrix and microenvironment that facilitates stem cell proliferation and differentiation through intercellular contact. (4) Indirect co-culture addresses the problem of insufficient sources of combined cells and immune rejection between individuals. The biological factors produced by the combined cells can be more easily made into clinical drugs and biological scaffold materials. (5) Though the co-culture system has superior performance in angiogenesis, there are few reports on other structures, and the regeneration of tissue from the co-culture outcome is uncertain. Furthermore, there are limitations to applying newly generated tissue generated by the co-culture system to clinical practice due to tooth anatomical constraints and rejection mechanisms. [ABSTRACT FROM AUTHOR]

Dental pulp stem cells (DPSCs) are a new type of mesenchymal stem cells (MSCs) found in the oral cavity with immunomodulation and tissue regeneration capacities. This study determined the impacts of nano-hydroxyapatite (nHA) prepared through Elaeagnus Angustifolia extract (EAE) to enhance the relative expression of immunomodulatory/dentin–pulp regeneration genes in DPSCs. To produce nHA and modified nHA via EAE (nHAEA), the sol–gel technique was used. The functional groups of nanoparticles (NPs), morphological, and optical features were determined using Fourier transform infrared (FTIR), X-ray diffraction (XRD), Scanning electron microscopy (SEM) together with energy-dispersive X-ray analysis (EDAX), and Transmission electron microscopy (TEM). The cell viability was then determined using the MTT method in the presence of various EAE, nHA, and nHAEA concentrations. Target gene expression was quantified using a real-time PCR procedure after treating DPSCs with an optimally non-toxic dose of EAE and NPs. The presence of the HA phase was reported with the XRD and FTIR results. According to the results of SEM and TEM, the rod-like NPs could be fabricated. nHAEAs were found to be characterized with low crystallite size, reduced diameter, lengthier, needle-like, and less agglomerated particles compared with nHA. The real-time PCR results demonstrated that nHAEA remarkably increased the expression of human leukocyte antigen-G5 (HLA-G5), vascular endothelial growth factor (VEGF), dentin sialophosphoprotein (DSPP), and interleukin6 (IL6) genes compared to the nHA group. These findings suggest that nHAEAs might have the potential application in the stemness capability of DPSCs for the treatment of inflamed/damaged pulp. [ABSTRACT FROM AUTHOR]

Abstract Background External root resorption is one of common complications of orthodontic treatment, while internal root resorption is rarely observed, and the difference between pulp and periodontal tissues during orthodontic treatment is still unknown. The purpose of this study was to evaluate the effects of orthodontic forces on histological and cellular changes of the dental pulp and periodontal tissues. Methods Orthodontic tooth movement model was established in Forty-eight adult male Wistar rats. The distance of orthodontic tooth movement was quantitatively analyzed. The histological changes of pulp and periodontal tissues were performed by hematoxylin–eosin staining, tartrate-resistant acid phosphate staining was used to analyze the changes of osteoclast number, immunohistochemistry analysis and reverse transcription polymerase chain reaction were used to examine the receptor of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) expression. The width of tertiary dentine was quantitatively analyzed. Tartrate-resistant acid phosphate staining and the erosion area of osteo assay surface plate was used to evaluate osteoclast activity. Results The orthodontic tooth movement distance increased in a force dependent manner, and reached the peak value when orthodontic force is 60 g. Heavy orthodontic force increased the RANKL expression of periodontal ligament srem cells (PDLSCs) which further activated osteoclasts and resulted in external root resorption, while the RANKL expression of dental pulp stem cells (DPSCs) was relatively low to activate osteoclasts and result in internal root resorption, and the dental pulp tend to form tertiary dentine under orthodontic force stimulation. Conclusions Heavy orthodontic forces activated osteoclasts and triggered external root resorption by upregulating RANKL expression in rat periodontal tissues, while there was no significant change of RANKL expression in dental pulp tissue under heavy orthodontic forces, which prevented osteoclast activation and internal root resorption.

The therapeutic effectiveness of dental pulp stem cells (DPSCs) is limited. Sirtuin 7 (SIRT7) has been reported to be associated with a variety of age-related diseases. We aimed to identify the regulatory role of SIRT7 in DPSC senescence and investigate the underlying mechanism. DPSCs were isolated from healthy adults, the stem markers were verified by flow cytomerty analysis. Replicative senescence was induced in DPSCs by serial passage and cells were analyzed at PD16 and 54. DPSC senescence was evaluated by observing senescence-associated β-galactosidase (SA-β-gal) and telomerase reverse transcriptase (TERT) activity. Meanwhile, the markers of senescence levels were monitored by western blotting assay. SIRT7 protein was pulled-down, and the binding relationship between SIRT7 and ROCK1 was verified by immunoprecipitation and western blotting methods. Replicative senescence was induced in DPSCs at PD54. The number of SA-β-gal stained DPSCs significantly increased in the PD54 group while the level of TERT activity was decreased. The cyclin-dependent kinase inhibitors p53, p21, and p16, which are markers of senescence, were markedly up-regulated at PD54. SIRT7 was also found to be lowly expressed at PD54. Inhibition of SIRT7 significantly accelerated the senescence of DPSCs. Moreover, SIRT7 can bind with ROCK1, and SIRT7 could lead to ROCK1 desuccinylation at K520. Inhibited ROCK1 significantly reversed the effects of SIRT7 knockdown on regulating DPSCs senescence. Our results demonstrate that the SIRT7/ROCK1 axis plays a key role in the regulation of DPSC senescence and provide a candidate target to improve the functional and therapeutic potential of DPSCs. • Identification of isolated DPSCs. • SIRT7 is lowly expressed during replicative senescence in DPSCs • Inhibition of SIRT7 enhances the senescence in DPSCs [ABSTRACT FROM AUTHOR]

Large bone tissue defects present a significant clinical challenge due to the lack of stem cells and an osteogenic microenvironment, leading to fibrotic healing and impaired bone regeneration. Microsphere-based cell-on three-dimensional (3D) culture systems show great promise for constructing osteogenic microtissues. However, the underlying mechanisms require further investigation. In this study, we propose a simple, scalable framework for highly efficient osteogenic microtissue construction, utilizing gelatin methacryloyl (GelMA) microspheres and dental pulp stem cells (DPSCs). The GelMA microspheres provide an extensive, scalable 3D framework for the autonomous adhesion, migration, and proliferation of DPSCs. Within the enormous 3D space created by the microspheres, DPSCs anchor to the microspheres and neighboring cells, inducing intrinsic tensile stress and simulating a mechanical force akin to "rock climbing training". Transcriptomic sequencing results reveal that the 3D spatial and mechanical microenvironment modulates biological processes involved in cell adhesion, extracellular matrix organization, and the positive regulation of cell migration. Further investigations demonstrate that triggering the FAK/YAP pathway mediate mechanical driven differentiation of DPSCs into the osteoblastic lineage in the excellent osteogenic microtissues. Moreover, this simple scalable 3D framework strategy is expected to enable the efficient and large-scale preparation of stem cell-based microtissues. [Display omitted] [ABSTRACT FROM AUTHOR]

Aims. By measuring the extent of cytokines secreted by human dental pulp stem cells (hDPSCs) from passages 2 through 10, the optimal passage of hDPSCs was determined. This offers a potential theoretical basis for the treatment of neurological disorders. Method. After isolation and culture of hDPSCs from human teeth, the morphological features of the cells were observed under an inverted microscope. hDPSCs were identified by their immunophenotypes and their multiple differentiation capability. Cytokine concentrations secreted in the supernatants at passages 2-10 were detected by ELISA. Results. hDPSCs were viewed as fusiform or polygonal in shape, with a bulging cell body, homogenized cytoplasm, and a clear nucleus. Moreover, they could differentiate into neuroblasts in vitro. hDPSCs at passage 3 were positive for CD29 (91.5%), CD73 (94.8%) and CD90 (96.7%), but negative for the hematopoietic markers CD34 (0.13%). ELISA results showed that hDPSCs at passage 3 had the highest secretion levels of vascular endothelial growth factor (VEGF), brain-derived neurotrophic factor (BDNF), and nerve growth factor (NGF), with the highest secretion level of Neurotrophin-3 (NT-3) being at passage 2. Conclusion. hDPSCs have steady biological features of stem cells and exhibit optimal proliferation potential. hDPSCs at different passages have different capacities in the secretion of VEGF, BDNF, NGF, and NT-3. In conclusion cytokines secreted by hDPSCs may prove to be appropriate in the treatment of neurological diseases.