Human Dental Pulp Research Articles

Human Dental Pulp Mesenchymal Stem Cell-Derived Soluble Factors Combined with a Nanostructured Scaffold Support the Generation of a Vascular Network In Vivo.

Among all strategies directed at developing new tools to support re-vascularization of damaged tissues, the use of pro-angiogenic soluble factors, derived from mesenchymal stem cells (MSCs), appears a promising approach for regenerative medicine. Here, we compared the feasibility of two devices, generated by coupling soluble factors of human dental pulp mesenchymal stem cells (DPSCs), with a nanostructured scaffold, to support angiogenesis once transplanted in mice. DPSCs were obtained from impacted wisdom tooth removal, usually considered surgical waste material. After 28 days, we verified the presence of active blood vessels inside the scaffold through optical and scansion electron microscopy. The mRNA expression of surface antigens related to macrophage polarization (CD68, CD80, CD86, CD163, CD206), as well as pro-angiogenic markers (CD31, CD34, CD105, Angpt1, Angpt2, CDH5) was evaluated by real-time PCR. Our results demonstrate the capability of DPSC-scaffold and DPSC soluble factors-scaffold to support angiogenesis, similarly to adipose stem cells, whereas the absence of blood vessels was found in the scaffold grafted alone. Our results provide evidence that DPSC-conditioned medium can be proposed as a cell-free preparation able to support angiogenesis, thus, providing a relevant tool to overcome the issues and restrictions associated with the use of cells.

Multifunctional Lithium-Doped Mesoporous Nanoparticles for Effective Dentin Regeneration in vivo.

Effective infection control without irritating the pulp tissue is the key to successful vital pulp therapy. Developing a novel antibacterial biomaterial that promotes dentin regeneration for pulp capping is thus a promising strategy for enhancing vital pulp therapy. Lithium-doped mesoporous nanoparticles (Li-MNPs) were synthesized using an alkali-catalyzed sol-gel method. The particle size, elemental distribution, surface morphology, pore structure, and ion release from Li-MNPs were measured. Human dental pulp stem cells (hDPSCs) and Streptococcus mutans (S. mutans) were used to evaluate the biological effects of Li-MNPs. In addition, a dental pulp exposure mouse model was used to evaluate the regenerative effects of Li-MNPs. Li-MNPs had a larger surface area (221.18 m2/g), a larger pore volume (0.25 cm3/g), and a smaller particle size (520.92 ± 35.21 nm) than MNPs. The in vitro investigation demonstrated that Li-MNPs greatly enhanced the biomineralization and odontogenic differentiation of hDPSCs through the Wnt/β-catenin signaling pathway. Li-MNPs showed a strong antibacterial effect on S. mutans. As expected, Li-MNPs significantly promoted dentin regeneration in situ and in vivo. Li-MNPs promoted dentin regeneration and inhibited S. mutans growth, implying a possible application as a pulp capping agent in vital pulp therapy.

Biological Function and Potential Applications of Garcinol in Human Dental Pulp Stem Cells

IntroductionThe regeneration of pulp tissue is crucial for true regenerative endodontic treatment, which requires a reduction in osteogenic differentiation. Garcinol, a histone acetyltransferase inhibitor, is a natural regulator that is known to suppress the osteogenic differentiation of dental pulp stem cells. In this study, the inhibitory effect of garcinol on the osteogenic differentiation of human dental pulp stem cells (hDPSCs) was evaluated using three-dimensional culture under in vitro and in vivo conditions. MethodshDPSCs were obtained from caries-free third molars and cultured with 10 μM garcinol for 7 days in an ultra-low attachment plate. The cell stemness and expression of osteogenic differentiation-related genes were analyzed using reverse transcription-polymerase chain reaction and single-cell analysis. A transplantation experiment was performed in mice to investigate whether garcinol-treated hDPSCs showed restrained osteogenic differentiation. ResultshDPSCs cultured in the U-shaped ultra-low attachment plate showed the highest expression of stemness-related genes. Garcinol-treated hDPSCs demonstrated downregulation of osteogenic differentiation, with lower expression of bone sialoprotein, which is related to bone formation, and higher expression of dentin sialophosphoprotein, which is related to dentin formation. However, the garcinol-treated hDPSCs did not show any alterations in their stemness. Consistent results were observed in the transplantation experiment in mice. ConclusionsGarcinol reduced the osteogenic differentiation of hDPSCs, which can contribute to true regenerative endodontic treatment.

Osteogenic differentiation enhancement of human dental pulp mesenchymal stem cells: A review

Background: The article aims to review osteogenic differentiation enhancement in human Dental Pulp Mesenchymal Stem Cells (hDPMSCs). Material and Methods: The literature review was carried out in PUBMED with the keywords: human Dental Pulp Mesenchymal Stem Cells (hDPMSCs) and osteogenic differentiation. Conclusions: Many research investigated the impact of different compounds on the osteogenic differentiation of hDPMSCs and the appliance by which these compounds promote osteogenesis in hDPMSCs which are a potential source for bone regeneration.

Development of cobalt-incorporated chitosan scaffold for regenerative potential in human dental pulp stem cells: An in vitro study

The aim of the study was to comparatively evaluate chitosan and Cobalt incorporated chitosan (CoCH) scaffold at varying concentrations in terms of their material characteristics, cytotoxicity and cell adhesion potential. In the present study, cobalt incorporated chitosan scaffolds at varying concentrations were prepared and dried. The synthesised scaffolds were characterised using XRD, FTIR, SEM-EDX and BET which revealed amorphous, porous surface of CoCH scaffolds and FTIR analysis showed the complexation confirming the chelation of cobalt with chitosan. The experimental scaffolds proved to be non-cytotoxic when compared to chitosan scaffolds on XTT analysis. Cell-seeding assay revealed enhanced adherence of hDPSCs to CoCH scaffold at 1:1 ratio in the concentration of 100 mL of 100 μmol/L cobalt chloride solution in 100mL of 2% chitosan solution, when compared to other groups. The results highlighted that 100 μmol/L concentration of cobalt chloride when incorporated in 1:1 ratio into 2 % CH solution yields a promising porous, biocompatible scaffold with enhanced cellular adhesion for dentin-pulp regeneration.

MicroRNA-4691-3p inhibits the inflammatory response by targeting STING in human dental pulp cells: A laboratory investigation.

The regulation of human dental pulp inflammation is not fully understood. This study aims to investigate the effect of miR-4691-3p on the cGAS-STING signalling cascade and its downstream cytokines production in human dental pulp cells (HDPCs). Normal dental pulp tissue and pulp tissue with irreversible pulpitis from third molars were collected. HDPCs were isolated from pulp tissue. The expression of STING mRNA and miR-4691-3p was measured by quantitative real-time PCR. Bioinformatic computation via TargetScanHuman 8.0 and a luciferase reporter assay was used to identify the targets of miR-4691-3p. A miR-4691-3p mimic and inhibitor were used to upregulate or downregulate miR-4691-3p expression in HDPCs. HDPCs were transfected with c-di-AMP, c-di-GMP, cGAMP, interferon stimulatory DNA (ISD) and bacterial genomic DNA. Immunoblot was performed to detect the phosphorylation of TBK1, p65 and IRF3. Enzyme-linked immunoassay was performed to detect the cytokines including IFN-β, TNF or IL-6 downstream of cGAS-STING. MiR-4691-3p expression was increased in human dental pulp tissue with irreversible pulpitis. Treatment of HDPCs using recombinant human IFN-β, TNF or IL-6 also upregulated miR-4691-3p. The bioinformatic prediction and luciferase reporter assay confirmed that STING was a direct target of miR-4691-3p. The miR-4691-3p mimic suppressed STING expression, the phosphorylation of TBK1, p65 and IRF3, and the IFN-β, TNF or IL-6 production. In contrast, the miR-4691-3p inhibitor enhanced the STING expression, the phosphorylation of TBK1, p65 and IRF3 and the IFN-β, TNF or IL-6 production. MiR-4691-3p negatively regulates the cGAS-STING pathway by directly targeting STING. This provides insight to utilize miRNA-dependent regulatory effect to treat endodontic disease as well as STING-dependent systemic inflammatory disease.

Repeat intramuscular transplantation of human dental pulp stromal cells is more effective in sustaining Schwann cell survival and myelination for functional recovery after onset of diabetic neuropathy

Background aimsMesenchymal stromal cell (MSC) therapy for diabetic neuropathy (DN) has been extensively researched in vitro and in pre-clinical studies; however, the clinical scenario thus far has been disappointing. Temporary recovery, a common feature of these studies, indicates that either the retention of transplanted cells deteriorates with time or recovery of supportive endogenous cells, such as bone marrow-derived MSCs (BM-MSCs), does not occur, requiring further replenishment. In DN, BM-MSCs are recognized mediators of Schwann cell regeneration, and we have earlier shown that they suffer impairment in the pre-neuropathy stage. In this study, we attempted to further elucidate the mechanisms of functional recovery by focusing on changes occurring at the cellular level in the sciatic nerve, in conjunction with the biodistribution and movement patterns of the transplanted cells, to define the interval between doses. Method & ResultsWe found that two doses of 1 × 106 dental pulp stromal cells (DPSCs) transplanted intramuscularly at an interval of 4 weeks effectively improved nerve conduction velocity (NCV) and restored motor coordination through improving sciatic nerve architecture, Schwann cell survival and myelination. Despite very minimal recovery of endogenous BM-MSCs, a temporary restoration of NCV and motor function was achieved with the first dose of DPSC transplantation. However, this did not persist, and a repeat dose was needed to consolidate functional improvement and rehabilitate the sciatic nerve architecture. ConclusionThus, repeat intramuscular transplantation of DPSCs is more effective for maintenance of Schwann cell survival and myelination for functional recovery after onset of DN.

LAMA5-inspired adhesive dodecapeptide facilitates efficient dentine regeneration: An in vitro and in vivo study.

The primary goal of this study was to investigate the potential effects of A5G81 in inducing reparative dentine (RD) formation both in vitro and in vivo. Cell adhesion was observed by crystal violet staining and quantified by Sodium Dodecyl Sulphate (SDS) extraction. Cell proliferation was investigated using Cell Counting Kit-8 (CCK-8) assay. Spreading of cytoskeleton was visualized using immunofluorescence staining. Protein expression level of Akt signalling pathway was compared in a human Akt pathway phosphorylation array. Genes that were up or downregulated by A5G81 were identified by RNA sequencing. The mRNA expression of odontoblastic markers was detected by quantitative real-time polymerase chain reaction (qPCR). Moreover, mineralization of human dental pulp cells (hDPCs) was visualized by alizarin red staining and quantified using cetylpyridinium chloride (CPC). A direct pulp-capping model was established in SD rats and the RD formation at 2 weeks after operation was observed using HE staining. A5G81 (optimal coating concentration: 0.5 mg/mL) promoted hDPCs adhesion and proliferation to a level that was similar to Type I collagen (COL-1). Meanwhile, A5G81 activated Akt signalling pathway, albeit to a lesser extent than COL-1. An inhibition test indicated that A5G81 induced hDPCs adhesion by activating PI3K pathway. A5G81 induced the expression of ECM remodelling genes and odontoblastic genes, which were demonstrated by RNA-seq and qPCR, respectively. In addition, A5G81 efficiently accelerated the mineralization of hDPCs in both immobilized and soluble forms, a property that makes it more applicable in dental clinic. Finally, the pulp-capping study in rats suggested that use of A5G81 could successfully induce the formation of RD within 2 weeks. Coating of A5G81 to non-tissue culture-treated polystyrene facilitates spreading, proliferation and differentiation of hDPCs, resulting in rapid RD formation in artificially exposed pulp.

Novel Bioactive Nanocomposites Containing Calcium Fluoride and Calcium Phosphate with Antibacterial and Low-Shrinkage-Stress Capabilities to Inhibit Dental Caries.

Composites are commonly used for tooth restorations, but recurrent caries often lead to restoration failures due to polymerization shrinkage-stress-induced marginal leakage. The aims of this research were to: (1) develop novel low-shrinkage-stress (L.S.S.) nanocomposites containing dimethylaminododecyl methacrylate (DMADDM) with nanoparticles of calcium fluoride (nCaF2) or amorphous calcium phosphate (NACP) for remineralization; (2) investigate antibacterial and cytocompatibility properties. Nanocomposites were made by mixing triethylene glycol divinylbenzyl ether with urethane dimethacrylate containing 3% DMADDM, 20% nCaF2, and 20% NACP. Flexural strength, elastic modulus, antibacterial properties against Streptococcus mutans biofilms, and cytotoxicity against human gingival fibroblasts and dental pulp stem cells were tested. Nanocomposites with DMADDM and nCaF2 or NACP had flexural strengths matching commercial composite control without bioactivity. The new nanocomposite provided potent antibacterial properties, reducing biofilm CFU by 6 logs, and reducing lactic acid synthesis and metabolic function of biofilms by 90%, compared to controls (p < 0.05). The new nanocomposites produced excellent cell viability matching commercial control (p > 0.05). Bioactive L.S.S. antibacterial nanocomposites with nCaF2 and NACP had excellent bioactivity without compromising mechanical and cytocompatible properties. The new nanocomposites are promising for a wide range of dental restorations by improving marginal integrity by reducing shrinkage stress, defending tooth structures, and minimizing cariogenic biofilms.

Bioenergetic profile and redox tone modulate in vitro osteogenesis of human dental pulp stem cells: new perspectives for bone regeneration and repair

BackgroundRedox signaling and energy metabolism are known to be involved in controlling the balance between self-renewal and proliferation/differentiation of stem cells. In this study we investigated metabolic and redox changes occurring during in vitro human dental pulp stem cells (hDPSCs) osteoblastic (OB) differentiation and tested on them the impact of the reactive oxygen species (ROS) signaling.MethodshDPSCs were isolated from dental pulp and subjected to alkaline phosphatase and alizarin red staining, q-RT-PCR, and western blotting analysis of differentiation markers to assess achievement of osteogenic/odontogenic differentiation. Moreover, a combination of metabolic flux analysis and confocal cyto-imaging was used to profile the metabolic phenotype and to evaluate the redox tone of hDPSCs.ResultsIn differentiating hDPSCs we observed the down-regulation of the mitochondrial respiratory chain complexes expression since the early phase of the process, confirmed by metabolic flux analysis, and a reduction of the basal intracellular peroxide level in its later phase. In addition, dampened glycolysis was observed, thereby indicating a lower energy-generating phenotype in differentiating hDPSCs. Treatment with the ROS scavenger Trolox, applied in the early-middle phases of the process, markedly delayed OB differentiation of hDPSCs assessed as ALP activity, Runx2 expression, mineralization capacity, expression of stemness and osteoblast marker genes (Nanog, Lin28, Dspp, Ocn) and activation of ERK1/2. In addition, the antioxidant partly prevented the inhibitory effect on cell metabolism observed following osteogenic induction.ConclusionsAltogether these results provided evidence that redox signaling, likely mediated by peroxide species, influenced the stepwise osteogenic expansion/differentiation of hDPSCs and contributed to shape its accompanying metabolic phenotype changes thus improving their efficiency in bone regeneration and repair.

Ruxolitinib altered IFN-β induced necroptosis of human dental pulp stem cells during osteoblast differentiation

ObjectiveThis study aimed to evaluate the role of ruxolitinib in the interferon beta (IFN-β) mediated osteoblast differentiation using human dental pulp stem cells (hDPSCs). DesignhDPSCs from five deciduous teeth of healthy patients were stimulated by adding human recombinant IFN-β protein (1 or 2 ng/ml) to the osteogenic differentiation induction medium. Substrate formation was determined using Alizarin Red staining, calcium concentration, and osteoblast marker expression levels. Ruxolitinib was used to inhibit the Janus kinase/signal transducers and activators of transcription (JAK-STAT) pathway. Apoptosis was detected using terminal deoxynucleotidyl nick-end labeling (TUNEL) staining, and necroptosis was detected using propidium iodide staining and phosphorylated mixed lineage kinase domain-like protein (pMLKL) expression. ResultsIn the IFN-β-treated group, substrate formation was inhibited by a reduction in alkaline phosphatase (ALP) expression in a concentration-dependent manner. Although the proliferation potency was unchanged between the IFN-β-treated and control groups, the cell number was significantly reduced in the experimental group. TUNEL-positive cell number was not significantly different; however, the protein level of necroptosis markers, interleukin-6 (IL-6) and pMLKL were significantly increased in the substrate formation. Cell number and ALP expression level were improved in the group administered ruxolitinib, a JAK-STAT inhibitor. Additionally, ruxolitinib significantly suppressed IL-6 and pMLKL levels. ConclusionRuxolitinib interfered with the IFN-β-mediated necroptosis and osteogenic differentiation via the JAK-STAT pathway.

The viability of human dental pulp cells and apical papilla cells after treatment with conventional calcium hydroxide and nanoparticulate calcium hydroxide at various concentrations

IntroductionNanoparticulate Ca(OH)2 had greater antibacterial effect than conventional Ca(OH)2. Conversely, a study reported that nanoparticulate Ca(OH)2 had toxicity against murine fibroblast. However, the study of nanoparticulate Ca(OH)2, involving human dental pulp cells (DPCs) and apical papilla cells (APCs) is lacking. The aim of this study is to compare the effects of conventional Ca(OH)2 and nanoparticulate Ca(OH)2 on the viability of DPCs and APCs. MethodsPrimary human DPCs/APCs from the 3rd to 5th passage were divided into control and experimental groups. In the control group, cells were cultured in complete media. In the experimental group, cells were cultured in complete media containing 10, 100, or 1000 μg/mL of either conventional Ca(OH)2 or nanoparticulate Ca(OH)2 for 1, 3, 5, and 7 days. After the treatment period, the cells were tested for viability using MTT assay. ResultsDPCs treated with conventional Ca(OH)2 in all concentrations at day 5 revealed significantly higher proliferation compared to nanoparticulate Ca(OH)2 treated groups. In additions, DPCs treated with 1000 µg/ml nanoparticulate Ca(OH)2 at day7 were significantly lower proliferation compared to DPCs treated with conventional Ca(OH)2. In contrast, APCs treated with 1000 µg/ml nanoparticulated Ca(OH)2 were significantly higher proliferation than APCs treated with 1000 µg/ml conventional Ca(OH)2 at day7. ConclusionsNanoparticulate Ca(OH)2 increased the viability of APCs and can be an alternative choice of intracanal medication for regenerative endodontic procedures. However, Nanoparticulate Ca(OH)2 exerted some effects on DPCs. The use of nanoparticulate Ca(OH)2 has no advantages over the conventional Ca(OH)2 for vital pulp therapy.

In vitro response of dental pulp stem cells to dural substitute grafts: Analysis of cytocompatibility and bioactivity.

The objective of this study was to assess and compare the cytocompatibility of decellularized porcine small intestine submucosal dural graft from Biodesign (BD) and polyester urethane-based Neuro-Patch (NP) dural substitute with the mineral trioxide aggregate (MTA) and the cyanoacrylate-based Histoacryl surgical adhesive. Furthermore, the study evaluated the inflammatory response and osteogenic differentiation of human dental pulp stem cells (hDPSCs) when cultured in direct contact with the dural substitutes in comparison with MTA. The viability of hDPSCs in direct contact with the tested materials was investigated in vitro by a CCK-8 assay. Additionally, the effects of dural substitutes and MTA on the expression of the inflammatory mediator interleukin-6 (IL-6) was investigated via enzyme-linked immunosorbent assay (ELISA), whilst effects on the differentiation were evaluated using alizarin red staining, alkaline phosphatase staining, ELISA and energy-dispersive X-ray elemental mapping. The dural substitutes were cytocompatible and promoted cellular adhesion. The Histoacryl and MTA demonstrated cytotoxicity in fresh preparations but showed a more favourable cellular reaction when set. Investigations of biological activity indicated that dural substitute membranes did not induce an inflammatory response or osteogenic differentiation of hDPSCs. In contrast, MTA induced the expression of IL-6 and alkaline phosphatase activity contributing to enhanced differentiation and mineralization. The dural substitute membranes showed cytocompatibility, did not provoke an inflammatory response and maintained the stemness of hDPSCs better than MTA. Additionally, the set Histoacryl surgical adhesive demonstrated good biocompatibility. Taken together, these results highlight the potential use of dural substitutes in regenerative endodontic procedures as coronal barriers alternative to MTA to reduce the incidence of intracanal calcifications.

Ultrasensitive two-photon probes for rapid detection of β-galactosidase during fruit softening and cellular senescence

Senescence is happening in every corner of the living organisms. β-galactosidase (β-gal) is one of the most important biomarkers during senescence in both plant and mammalian cells. Most β-gal fluorescent probes were focused on bio-imaging, only a few probes were developed for the detection of β-gal in fruit, and the probes that could detect β-gal in both fruits and living cells were even less. Here, two β-gal probes (TNap-βGal and TBNap-βGal) were synthesized, which can not only image the increase of β-gal during both fruits softening and cellular senescence, but also prove that bananas are not suitable for storage in refrigerator and the subsequent accumulation of β-gal still in lysosome of mammalian cells. In addition, TNap-βGal was successfully applied to two-photon imaging of endogenous β-gal in both hDPMSCs and tissues of human dental pulp for the first time.

Micropattern‐Based Stem Cell Gym: Mechanical Memory Enhanced Stemness Maintenance of Human Dental Pulp Stem Cells and Nerve Regeneration

AbstractDental pulp stem cells (DPSCs) which are derived from the neural crest are promising tools in stem cell therapy in the clinic. However, they tend to lose their multi‐lineage potency spontaneously during long‐term expansion in vitro. A micropattern‐based system is developed to train DPSCs with 1024 µm2 and aspect ratios of 1:2 and 1:4, which enhances their stemness and differentiation potential. This mechanical memory induced by micropattern would also rescue the pluripotency of long‐term expansion DPSCs. Micropatterns remodel the cytoskeleton and nuclear morphology, soften the cells, and induce the mitochondria into an immature state. Micropattern also activates extracellular related kinase, signal transducer, and activator of transcription 3, and inhibits Yes‐associated protein nuclear localization to enhance the stemness of DPSCs. In vivo implantation proves that DPSCs mechanically trained by micropattern could promote nerve regeneration in rats with peripheral nerve injury. A micropattern‐based approach is provided as a stem cell gym that absents exogenous growth factors to enhance the stemness and regenerative properties of DPSCs. It may shed light on future stem cell therapy.

Integration of blue light with near-infrared irradiation accelerates the osteogenic differentiation of human dental pulp stem cells.

Blue light is used less in photobiomodulation than red or near-infrared light because of concerns about its high energy. However, some reports have suggested that blue light releases NO from nitrosated proteins, affects cell signal regulation, and promotes stem cell differentiation. Because blue and red lights could have different mechanisms of action, their combination is expected to have new consequences. In this study, human dental pulp stem cells (hDPSCs) were sequentially exposed to blue and near-infrared light to study their effects on proliferation, osteogenic differentiation, and immunomodulation. We found that NIR irradiation applied after blue light can reduce blue light toxicity improving the cell viabiltiy. Delayed luminescence and transmission electron microscopy studies showed that this combination excited hDPSCs and activated mitochondrial biogenesis. Those modulations accelerated hDPSC differentiation, as shown by an increase of about 1.3-fold in alkaline phosphatase activity in vitro and an about 1.5-fold increase in the osteocalcin-positive regions in cells implanted in nude mice compared with mice exposed to near-infrared alone.

Improving Pure Titanium's Biological and Mechanical Characteristics through ECAP and Micro-Arc Oxidation Processes.

Pure titanium is limited to be used in biomedical applications due to its lower mechanical strength compared to its alloy counterpart. To enhance its properties and improve medical implants feasibility, advancements in titanium processing technologies are necessary. One such technique is equal-channel angular pressing (ECAP) for its severe plastic deformation (SPD). This study aims to surface modify commercially pure titanium using micro-arc oxidation (MAO) or plasma electrolytic oxidation (PEO) technologies, and mineral solutions containing Ca and P. The composition, metallography, and shape of the changed surface were characterized using X-ray diffraction (XRD), digital optical microscopy (OM), and scanning electron microscope (SEM), respectively. A microhardness test is conducted to assess each sample's mechanical strength. The weight % of Ca and P in the coating was determined using energy dispersive spectroscopy (EDS), and the corrosion resistance was evaluated through potentiodynamic measurement. The behavior of human dental pulp cell and periodontal cell behavior was also studied through a biomedical experiment over a period of 1-, 3-, and 7-days using culture medium, and the cell death and viability can be inferred with the help of enzyme-linked immunosorbent assay (ELISA) since it can detect proteins or biomarkers secreted by cells undergoing apoptosis or necrosis. This study shows that the mechanical grain refinement method and surface modification might improve the mechanical and biomechanical properties of commercially pure (CP) titanium. According to the results of the corrosion loss measurements, 2PassMAO had the lowest corrosion rate, which is determined to be 0.495 mmpy. The electrode potentials for the 1-pass and 2-pass coated samples are 1.44 V and 1.47 V, respectively. This suggests that the coating is highly effective in reducing the corrosion rate of the metallic CP Ti sample. Changes in the grain size and the presence of a high number of grain boundaries have a significant impact on the corrosion resistance of CP Ti. For ECAPED and surface-modified titanium samples in a 3.6% NaCl electrolyte solution, electrochemical impedance spectroscopy (EIS) properties are similar to Nyquist and Bode plot fitting. In light of ISO 10993-5 guidelines for assessing in vitro cytotoxicity, this study contributes valuable insights into pulp and periodontal cell behavior, focusing specifically on material cytotoxicity, a critical factor determined by a 30% decrease in cell viability.

The Secretome of Human Dental Pulp Stem Cells and Its Components GDF15 and HB-EGF Protect Amyotrophic Lateral Sclerosis Motoneurons against Death.

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.

Dextrin hydrogel loaded with a macroporous Bonelike® scaffold and dental pulp stem cells for critical-sized defect repair

Regeneration of severe bone defects remains a challenge. A formulation of synthetic glass-reinforced hydroxyapatite bone substitute, Bonelike® Poro (BL®P), 250–500 µm-diameter, with a dextrin-based hydrogel (HG), further loaded with human dental pulp stem cells (hDPSCs) with osteogenic differentiation ability, was tested for the management of critical-sized defects in an ovine model.Morphology, calcium release, and mechanical strength of HG + BL®P were analyzed. Then, BL®P, HG + BL®P, and 106 hDPSCs-loaded HG + BL®P were implanted in ovine critical-sized 14 mm-diameter calvaria defects. Bone samples were collected after 3 and 6 weeks for histological and micro-CT analysis.BL®P exhibits a suitable porous size for cell ingrowth, from the nm (>200 nm) to the µm (5 µm) range. The addition of BL®P granules to the HG resulted in increased compressive elastic modulus and ultimate tensile strength. The mildly acidic nature of the HG contributed to a faster dissolution of granules. In vivo results confirmed the HG suitability as a carrier, providing better defect filling, easy handling, and injectability of BL®P without compromising new bone formation nor biocompatibility. The HG + BL®P formulations can successfully regenerate critical-sized defects; however, addition of hDPSCs did not significantly enhance new bone formation under these conditions.Granular BL®P provides an effective alternative to autologous grafts. The HG acts as a biocompatible carrier of granular bone substitutes and cells, conferring injectability and cohesivity.

Conditioned Media from Human Pulp Stem Cell Cultures Improve Bone Regeneration in Rat Calvarial Critical-Size Defects.

The aim of this study was to test whether lyophilized conditioned media from human dental pulp mesenchymal stem cell cultures promote the healing of critical-size defects created in the calvaria of rats. Prior to the surgical procedure, the medium in which dental pulp stem cells were cultured was frozen and lyophilized. After general anesthesia, an 8 mm diameter bone defect was created in the calvaria of twenty-four rats. The defects were filled with the following materials: xenograft alone (G1) or xenograft associated with lyophilized conditioned medium (G2). After 14 or 42 days, the animals were euthanized, and the specimens processed for histologic and immunohistochemical analysis. Bone formation at the center of the defect was observed only in the G2 at 42 days. At both timepoints, increased staining for VEGF, a marker for angiogenesis, was observed in G2. Consistent with this, at 14 days, G2 also had a higher number of blood vessels detected by immunostaining with an anti-CD34 antibody. In conclusion, conditioned media from human dental pulp mesenchymal stem cell cultures had a positive effect on the regenerative process in rat critical-size bone defects. Both the formation of bone and enhancement of vascularization were stimulated by the conditioned media.