Search Results (60)

Autologous tooth-derived grafts are increasingly being investigated for bone regeneration, as dentin shares with bone a mineral phase and an organic matrix rich in type I collagen and non-collagenous proteins. Deciduous teeth are particularly attractive as biomaterials because they are easily obtained after physiological exfoliation, without additional surgical harvesting or donor-site morbidity and may expose a protein-rich matrix after processing. Whether deciduous teeth retain a biologic advantage after prolonged dry storage remains poorly documented. This proof-of-concept ex vivo and in vitro study compared pooled deciduous teeth from six different donors (exfoliated at least 10 years before the experiment and stored dry at room temperature conditions) with six freshly extracted third molars. The teeth were ground using a dedicated device, and conditioned supernatants were collected at 72 h (T1) and 28 days (T2). Osteocalcin, osteonectin, and BMP-2 were quantified by ELISA, and T1 supernatants were applied to human primary osteoblasts to assess the osteogenic response using qRT-PCR and immunofluorescence. Deciduous teeth-conditioned supernatants showed higher osteocalcin and osteonectin release than permanent teeth at both time points, whereas BMP-2 levels were comparable, though with higher values in deciduous samples. In osteoblasts, deciduous teeth-conditioned supernatants induced enhanced osteogenic responses, including greater activation of Collagen I, Osterix, RUNX-2, Osteocalcin, BMP-2 genes, and higher expression of bone-related proteins. Within the limits of this exploratory study, dry-stored deciduous teeth preserved a biologically active dentin matrix and showed a more favorable osteogenic profile than freshly extracted permanent teeth, supporting further investigation into standardized storage protocols and their potential use in regenerative applications. Full article

Stem cell heterogeneity represents a critical yet underexplored variable in laser-assisted regenerative strategies. While photobiomodulation has been shown to influence mesenchymal stem cell (MSC) behavior, it remains unclear whether stem cell maturation status modulates responsiveness to Er,Cr:YSGG irradiation. This study investigated the differential response of magnetically separated STRO-1⁺ and STRO-1⁻ SHED subpopulations to low-power Er,Cr:YSGG laser stimulation (0.10 W and 0.25 W), focusing on viability, extracellular matrix production, and lineage commitment. STRO-1⁺ cells comprised 13.4% ± 1.2% of the total Stem Cells from Human Exfoliated Deciduous teeth (SHED) population. Laser exposure did not impair metabolic activity in either subpopulation. Collagen synthesis demonstrated a power- and time-dependent increase, with maximal enhancement observed in STRO-1⁺ cells at 0.25 W after 7 days. Laser irradiation selectively promoted osteogenic differentiation, as evidenced by increased alkaline phosphatase (ALP) expression at 0.10 W and enhanced mineral deposition, while chondrogenic potential remained unaffected and adipogenesis was reduced following 0.10 W exposure. These findings suggest that ALP expression is temporally and power-dependently modulated during osteogenic progression. Overall, Er,Cr:YSGG photobiomodulation does not uniformly affect heterogeneous SHED populations but modulates lineage allocation and extracellular matrix deposition in a maturation- and power-dependent manner. Integrating stem cell subpopulation selection with laser-based bioactivation may represent a strategy to refine regenerative endodontic and biomaterial-guided therapies. Full article

Urethral stricture is a disease of fibrotic narrowing that compromises the urethral mucosa and spongiosum. Oral mucosal graft urethroplasty delivers excellent outcomes in complex cases, yet its procedural demands restrict availability beyond specialized centers. Endoscopic transplantation of oral mucosa has been proposed; while feasibility is shown, clinical efficacy remains suboptimal. We asked whether extracellular vesicles from stem cells of human exfoliated deciduous teeth (SHED-EVs) promote oral mucosa fibroblast (OMF) growth under urethra-mimetic paracrine conditions and whether culture growth phase tunes EV function. SHED-EVs were collected during logarithmic (SHED-EV-L) or stationary (SHED-EV-S) phases under xeno-free conditions, isolated by a standardized workflow, and characterized by nanoparticle tracking analysis. miRNA cargo was profiled with a human miRNA microarray platform and normalized for comparative analyses. OMF proliferation was quantified in a horizontal indirect co-culture with urethral epithelial cells using incubator-based time-lapse imaging. SHED-EV-L produced a sustained pro-proliferative effect across 24–96 h, whereas SHED-EV-S showed a weaker early effect with a late catch-up; both exceeded vehicle at 96 h. Fibrosis-related miRNA heat maps showed culture growth phase-dependent patterns: SHED-EV-L displayed relatively higher signals for miR-31-3p, miR-146b-3p, several let-7 members, and selected miR-181 isoforms, whereas SHED-EV-S showed a marked relative increase of miR-486-3p; miR-21, miR-99/100, and miR-205 were broadly comparable between phases. These findings indicate that culture growth phase is a practical design lever that orients SHED-EV cargo and function, supporting phase-matched formulations for adjunctive transurethral applications and motivating in vivo validation and manufacturing-oriented quality controls. Full article

Mesenchymal stem cells derived from human exfoliated deciduous teeth (SHED) are a promising source for regenerative therapies due to their multipotency, proliferative capacity, and immunomodulatory properties. The present study aimed to isolate and characterize SHED subpopulations based on CD71 and CD146 expression and evaluate their multilineage differentiation potential. SHED were obtained from pediatric donors and separated into CD71⁺, CD71⁻, CD146⁺, and CD146⁻ fractions using magnetic-activated cell sorting (MACS). CD71⁺/CD71⁻ and CD146⁺/CD146⁻ populations were isolated independently; no simultaneous double sorting for both markers was performed. Immunocytochemistry was employed to confirm the expression of surface and intracellular markers, including STRO-1, CD44, nestin, and vimentin. Multilineage differentiation assays toward osteogenic, adipogenic, and chondrogenic lineages revealed that CD71⁺ cells exhibited reduced osteogenic capacity compared to CD71⁻ cells, whereas CD146⁺ cells showed enhanced osteogenic and adipogenic differentiation. Chondrogenic differentiation seemed unaffected by marker expression under the 2D conditions employed. These results highlight functional heterogeneity within SHED populations and indicate that CD71 and CD146 independently influence differentiation outcomes. The selective enrichment of CD146⁺ SHED may enhance osteogenic and adipogenic regenerative applications, while CD71⁺ subsets may serve as a valuable model for studying proliferation and paracrine effects. Limitations include the use of in vitro differentiation assays and the absence of in vivo validation; additionally, combined CD71/CD146 analysis may further clarify the relationship between metabolic activity and stem/progenitor niche characteristics. Overall, marker-based characterization of SHED subpopulations provides insight into their biological properties and potential utility in targeted cell-based therapeutic strategies. Full article

Urethral stricture involves fibrotic narrowing of the urethral mucosa and spongiosum. Although urethroplasty using oral mucosal grafts is the gold standard for complex cases due to its high success rate, technical complexity limits its broader adoption. To address this, endoscopic transplantation of oral mucosal tissue has been proposed. While feasibility has been demonstrated, clinical efficacy remains suboptimal. Developing adjunctive factors that facilitate mucosal engraftment may improve outcomes of endoscopic transplantation. Extracellular vesicles (EVs)—membrane-bound nanoparticles secreted by cells that deliver miRNAs and other bioactive molecules—have recently emerged as promising candidates. We investigated EVs derived from four mesenchymal stromal cell (MSC) sources—stem cells from human exfoliated deciduous teeth (SHED), adipose tissue, umbilical cord, and bone marrow (BM)—isolated during both logarithmic (log) and stationary culture phases. miRNA profiling revealed distinct phase- and origin-specific signatures. SHED-derived EVs from the log phase and bone marrow-derived EVs from the stationary phase expressed miR-31, the let-7 family, and miR-205, suggesting early wound healing potential. In contrast, stationary-phase SHED-EVs and log-phase BM-MSC-EVs were enriched in the miR-99 family and miR-31, indicating potential roles in epithelial stabilization and fibrosis modulation. These findings support phase-specific application of MSC-EVs to optimize mucosal engraftment in transurethral reconstruction. Full article

Background: Acute kidney injury (AKI) is a severe condition with high morbidity and mortality. Mesenchymal stem cells (MSCs) and their conditioned medium (MSCs-CM), containing extracellular vesicles (EVs), have shown therapeutic potential. This study evaluates the effects of stem cells from human exfoliated deciduous teeth (SHED) and their conditioned medium (SHED-CM) on cisplatin-induced AKI in an in vivo model. Methods: Male Wistar rats (n = 21) were randomly divided into three groups: a control group (cisplatin-induced AKI without intervention), a SHED group (receiving intraperitoneal SHED injections), and a SHED-CM group (receiving intraperitoneal SHED-CM injections). Kidney weight, urine parameters, histopathology, and injury scores were assessed. Results: SHED-CM treatment significantly improved renal function, with reduced kidney injury scores (1.42 ± 0.18 vs. 2.57 ± 0.23 in the control, p < 0.05). Kidney weight was lower in the SHED-CM group compared to the control (1.12 ± 0.05 g vs. 1.36 ± 0.07 g, p < 0.05). SHED-CM also significantly increased urine volume (2.41 ± 0.19 mL vs. 1.23 ± 0.14 mL in the control, p < 0.01) and reduced urinary protein concentration (27.3 ± 2.1 mg/dL vs. 48.5 ± 3.4 mg/dL in the control, p < 0.05). Histopathological analysis revealed reduced tubular necrosis and inflammation, with SHED-CM showing superior improvement over SHED. Conclusions: SHED-CM demonstrated superior therapeutic efficacy over SHED injection in mitigating cisplatin-induced AKI, likely due to extracellular vesicle-mediated paracrine effects. These findings support SHED-CM as a promising, non-invasive alternative for MSC-EV-based AKI therapy. Full article

Background: This study aimed to evaluate the cytotoxicity of bulk-fill composite materials compared to conventional compomers on stem cells from human exfoliated deciduous teeth. Methods: 90 standardized resin composite discs (4 mm thick, 4 mm diameter) were fabricated using a 3D-printed plate, comprising four bulk-fill composites (SDR, Tetric EvoCeram Bulk-Fill, VisCalor Bulk, Cention-N) and one compomer (Dyract XP). Samples were polymerized per the manufacturer’s instructions and sterilized. Stem cells were isolated from the pulp of exfoliated primary teeth. Cells were cultured and exposed to extracts of the composite materials soaked in culture medium for 24 h. Cytotoxicity was assessed using the MTT colorimetric assay, measuring cell viability via mitochondrial activity, and the Annexin V assay, quantifying apoptosis and necrosis via flow cytometry. Statistical analysis was performed using ANOVA and Tukey post hoc tests. Results: All materials significantly reduced cell viability compared to the control (p < 0.05), with optical density values indicating high cytotoxicity. Tetric EvoCeram exhibited the lowest necrosis and apoptosis levels, while Dyract XP showed the highest necrosis. Statistical analysis revealed no significant cytotoxicity differences among most bulk-fill composites (p < 0.05). Conclusion: Bulk-fill composites and conventional compomer tested exhibit comparable and significant cytotoxic effects on stem cells from human exfoliated primary teeth pulp. While these materials offer clinical advantages in pediatric dentistry due to ease and speed of application, their use underscores the dilemma of balancing operative efficiency with biological safety, and their cytotoxic profiles should be taken into consideration prior to application. Full article

Background/Objectives: Alzheimer’s disease (AD), a neurodegenerative condition, produces dementia and cognitive debility. Lately, preclinical models of AD have shown neuroregenerative potential of mesenchymal stem cells of dental origin (DMSC). This scoping review aims to map and synthesize the evidence on the therapeutic applications of DMSCs in AD management. Methods: This review followed the Arksey and O’Malley framework for scoping reviews and PRISMA-ScR guidelines. Scopus, Medline (Pubmed), Web of Science, and Embase databases were searched for published literature until February 2025. Data was mapped according to the type of DMSC and their therapeutic properties useful in AD management, like neuro differentiation, neuroprotection through increased neuron number and vitality, anti-neuroinflammation, mitochondrial repair, and improved cognition. Results: A total of 22 articles were included. A research gap existed, as most studies were preclinical (in vitro and animal models) with no clinical trials in humans. Furthermore, they mostly evaluated neuroregenerative properties of dental pulp stem cells (DPSC) and stem cells from human-exfoliated deciduous teeth (SHED), while Periodontal ligament stem cells (PDLSC) were least studied. All the DMSCs were neuroprotective and increased neuron number and vitality. Neurodifferentiation was reported in DPSCs and PDLSCs, while DPSCs and SHEDs showed anti-neuroinflammation, mitochondrial repair, and improved cognition in AD animal models. Conclusions: Although the DPSCs and SHEDs showed promising outcomes in preclinical models of AD, a gap exists as results have not been translated into human clinical trials. Future advances may identify plausible ways of applying the DMSCs to regain the lost neurons and re-establish a healthy brain microenvironment. Full article

The tissue regenerative potential of the liquid component of mesenchymal stem cells has gained significant attention. Stem cells from human exfoliated deciduous teeth-conditioned medium (SHED-CM), which is often extracted during orthodontic treatment, promotes bone regeneration. However, further investigation is warranted to determine which component of SHED-CM affects bone regeneration. Therefore, we focused on the extracellular vesicles contained in SHED-CM (SHED-EVs) and aimed to study their effects on osteoblasts. SHED-EVs were isolated using a pellet-down EV extraction kit and identified using transmission electron microscopy and NanoSight. SHED-EVs were added to human calvarial osteoblasts (HCOs), and cell proliferation and migration ability were examined with Incucyte^® and BrdU. Alkaline phosphatase (ALP) expression was confirmed using real-time PCR and ALP quantification. The bone differentiation potential was examined using Alizarin Red S (ARS) staining. SHED-EVs promoted proliferation and migration of HCOs. Real-time PCR and ALP quantification results demonstrated that HCOs cultured with SHED-EVs exhibited increased ALP expression. ARS staining revealed that SHED-EVs promoted bone differentiation of HCOs. These results suggest that SHED-EVs promote cell proliferation and migration and bone regeneration of osteoblasts, highlighting their potential in the development of bone regeneration therapies. Full article

Background/Objectives: Metabolic dysfunction-associated steatohepatitis (MASH), characterized by liver inflammation, fibrosis, and fat accumulation, can develop into cirrhosis and liver cancer. Despite its increasing prevalence worldwide, there are few established therapies for advanced MASH. We previously demonstrated that stem cells from human exfoliated deciduous teeth-conditioned media (SHED-CM) exerted therapeutic effects in a MASH mouse model. The gut–liver axis is thought to be associated with liver disease progression, and soluble Siglec-9 (sSiglec-9), an immunoinhibitory receptor, is a key protein in SHED-CM that induces anti-inflammatory macrophages and has intestinal epithelial protective effects. Therefore, we evaluated sSiglec-9’s role in intestinal barrier protection in MASH mice. Methods: We evaluated sSiglec-9 effects on intestinal barrier function using in vitro Caco-2 cell monolayers injured by TNF-α and IFN-γ. For the MASH mouse model, male C57BL/6J mice were given a Western diet and high-sugar solution orally; to induce liver injury, CCl4 was intraperitoneally administered for 12 weeks. Mice were treated weekly with 10 ng/g sSiglec-9 or vehicle. Intestinal permeability was assessed by blood 4 kDa FITC-dextran concentration, and intestinal transcriptomes and liver histology were analyzed. Results: sSiglec-9 decreased intestinal permeability and liver inflammation in MASH mice. sSiglec-9 and SHED-CM reduced 4 kDa FITC-dextran permeability in injured Caco-2 cells, and sSiglec-9 significantly reduced intestinal permeability and modulated expression of 34 intestinal genes. The NAFLD Activity Score indicated significantly reduced inflammation following sSiglec-9 treatment. Conclusions: sSiglec-9 may protect intestinal barrier function by mitigating mucosal inflammation. sSiglec-9 treatment may represent a novel therapeutic approach for MASH via gut–liver axis modulation. Full article

This study aimed to explore how amelogenin can improve stem cells from human exfoliated deciduous teeth (SHED)–based bone regeneration and promote tissue healing as a treatment for critical-sized bone defects. SHED was induced into bone differentiation by using osteogenic differentiation medium. Real-time polymerase chain reaction, alkaline phosphatase (ALP) staining and quantification, and Alizarin Red S staining, as well as calcium and osteocalcin quantification were performed to assess differentiation. On day 18, a significant increase was observed in the expression of RUNX2, CBFB, BGLAP, COL1, BMP2, BMP4, NOTCH1, NOTCH2, and NES. Osteocalcin gene expression continued to increase significantly. ALP activity was significantly higher in the amelogenin-treated group than in the control group on days 7, 10, and 14. On day 14, enhanced ALP staining was observed in the amelogenin-treated group. Calcium and osteocalcin levels were significantly higher in the amelogenin-treated group than in the control group on day 21. This study suggests that combining SHED and amelogenin may be effective for bone regeneration, offering a potential new approach in regenerative medicine. Full article

Supernumerary tooth-derived pulp stem cells (SNTSCs) hold promise for endodontic regeneration, yet little is known about the similarities and diversities of SNTSCs relative to other dental-derived mesenchymal stem cells. Herein, we compare the biological characteristics of SNTSCs with dental pulp stem cells (DPSCs) and stem cells from human exfoliated deciduous teeth (SHED). Cell proliferation, migration, and odontogenic differentiation potential, as well as viability and aging-related phenotype after long-term storage, were evaluated. Additionally, gene expressions during induced odontogenic differentiation were profiled by transcriptome sequencing. Our findings indicated that the SNTSCs outperformed the DPSCs but were inferior to the SHED in cell proliferation. The SNTSCs exhibited comparable migratory capacity to the SHED and surpassed the DPSCs. Of particular interest, the odontogenic differentiation potential followed the pattern of SHED > SNTSCs > DPSCs. After two years of storage, the SNTSCs showed weakness in resistance to apoptosis induced by lipopolysaccharide, whereas difference between the SNTSCs and SHED in stemness and senescence was not obvious. Transcriptome analysis revealed that upregulated genes in the SNTSCs were particularly enriched in inflammatory signaling pathways compared to both the DPSCs and SHED. Collectively, SNTSCs share many satisfactory features in proliferation and differentiation with SHED, which may serve as a promising alternative cell source for endodontic regeneration. Full article

SHEDs have demonstrated significant potential in cell therapy due to their superior proliferation rate, self-renewal and differentiation capacity (particularly neurogenesis attributed to their neural crest origin), and the less invasive procedure required for tissue collection compared to other stem cells. However, there is no established criterion to verify the minimum qualification to select one from numerous candidates, especially for SHEDs’ cultured FBS-free medium for clinic application. For that, we performed a characteristic analysis containing the growth rate, colony-forming unit (CFU) number, average colony size, and migration capacity with hPL-cultured SHEDs from 21 different donors, and we suggest the result as a minimum standard to filter out unqualified candidates. In addition, in the secretome analysis to predict the paracrine effect, it was found that upregulated proteins compared to the control were related to angiogenesis, immune response, and BMP signaling, and this was found to have a strong correlation only with protein concentration. This study presents a minimum standard for selecting cell therapy candidates and suggests the protein concentration of a conditioned medium as a cost-effective tool to expect the paracrine effect of SHEDs. Full article

This study aimed to evaluate the effects of CD146, a surface antigen of mesenchymal stem cells from human exfoliated deciduous teeth (SHEDs), on angiogenic potential. SHEDs were isolated from patients’ deciduous teeth and sorted into CD146-positive (CD146 + SHED) and CD146-negative (CD146 − SHED) populations. Three groups—non-sorted SHED, CD146 + SHED, and CD146 − SHED—were compared. Angiogenic potential was assessed by co-culturing each group with human umbilical vein endothelial cells (HUVECs) and evaluating lumen formation using an endothelial tube formation assay. The gene and protein expression levels of angiogenic markers, including vascular endothelial growth factor (VEGF), VEGF receptor 2 (VEGFR2), cluster of differentiation 31 (CD31), and basic fibroblast growth factor (bFGF), were analyzed using a real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The tube formation assay revealed significantly enhanced angiogenic potential in CD146 + SHED and non-sorted SHED compared to CD146 − SHED. The gene and protein expression levels of VEGF, VEGFR2, CD31, and bFGF were significantly upregulated in CD146 + SHED and non-sorted SHED, highlighting superior angiogenic capabilities in CD146 + SHED. CD146 + SHED demonstrated enhanced angiogenic potential compared to CD146 − SHED, supporting their use in regenerative therapies targeting angiogenesis. Full article

Three dimensional (3D) printing materials are widely used in dental applications, but their biocompatibility and interactions with human cells require evaluation. This study aimed to identify materials meeting biocompatibility, mechanical strength, and tissue-forming requirements for safe dental applications. We assessed the cytotoxicity of resins and thermoplastic filaments in human HaCaT keratinocytes, gingival fibroblasts (hGFs), and stem cells from human exfoliated deciduous teeth (SHED) using PrestoBlue assays. Three resins, including two types of surgical guide resins, exhibited strong cytotoxicity after 4–72 h, while 2 h exposure to an FDA-approved surgical guide resin did not affect SHED cell viability. In contrast, six thermoplastic filaments showed no significant cytotoxicity even after 72 h. Among these, polycarbonate–acrylonitrile butadiene styrene (PC-ABS) demonstrated excellent toughness, heat resistance, and surface quality at a low cost. SHED cells cultured on PC-ABS dishes and micro bone structures showed strong proliferation and osteogenic potential. Culture inserts made of PC-ABS also supported the growth of HaCaT keratinocytes and the hGFs formed gingival tissue, which was superior to that formed on commercially available PET inserts. In conclusion, PC-ABS is a promising 3D printing material for dental applications due to its biocompatibility, ability to promote osteogenesis, and support for gingival tissue formation, with no observed cytotoxicity. Full article