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Biomedical Materials (Bristol, England) Jun 2024Cell-based tissue engineering often requires the use of scaffolds to provide a three-dimensional (3D) framework for cell proliferation and tissue formation....
Cell-based tissue engineering often requires the use of scaffolds to provide a three-dimensional (3D) framework for cell proliferation and tissue formation. Polycaprolactone (PCL), a type of polymer, has good printability, favorable surface modifiability, adaptability, and biodegradability. However, its large-scale applicability is hindered by its hydrophobic nature, which affects biological properties. Composite materials can be created by adding bioactive materials to the polymer to improve the properties of PCL scaffolds. Osteolectin is an odontogenic factor that promotes the maintenance of the adult skeleton by promoting the differentiation of LepR+ cells into osteoblasts. Therefore, the aim of this study was to evaluate whether 3D-printed PCL/osteolectin scaffolds supply a suitable microenvironment for the odontogenic differentiation of human dental pulp cells (hDPCs). The hDPCs were cultured on 3D-printed PCL scaffolds with or without pores. Cell attachment and cell proliferation were evaluated using EZ-Cytox. The odontogenic differentiation of hDPCs was evaluated by alizarin red S staining and alkaline phosphatase assays. Western blot was used to evaluate the expression of the proteins DSPP and DMP-Results: The attachment of hDPCs to PCL scaffolds with pores was significantly higher than to PCL scaffolds without pores. The odontogenic differentiation of hDPCs was induced more in PCL/osteolectin scaffolds than in PCL scaffolds, but there was no statistically significant difference. 3D-printed PCL scaffolds with pores are suitable for the growth of hDPCs, and the PCL/osteolectin scaffolds can provide a more favorable microenvironment for the odontogenic differentiation of hDPCs.
Topics: Humans; Dental Pulp; Polyesters; Printing, Three-Dimensional; Tissue Scaffolds; Cell Differentiation; Odontogenesis; Cell Proliferation; Tissue Engineering; Cells, Cultured; Biocompatible Materials; Cell Adhesion; Osteoblasts
PubMed: 38740059
DOI: 10.1088/1748-605X/ad4ad9 -
Biomedical Materials (Bristol, England) Jun 2024This study aimed to investigate the effects of magnesium-doped bioactive glass (Mg-BG) on the mineralization, odontogenesis, and anti-inflammatory abilities of human...
This study aimed to investigate the effects of magnesium-doped bioactive glass (Mg-BG) on the mineralization, odontogenesis, and anti-inflammatory abilities of human dental pulp stem cells (hDPSCs). Mg-BG powders with different Mg concentrations were successfully synthesized via the sol-gel method and evaluated using x-ray diffraction, Fourier-transform infrared spectroscopy, scanning electron microscopy, and transmission electron microscopy. Apatite formation was observed on the surfaces of the materials after soaking in simulated body fluid. hDPSCs were cultured with Mg-BG powder extracts in vitro, and no evident cytotoxicity was observed. Mg-BG induced alkaline phosphatase (ALP) expression and mineralization of hDPSCs and upregulated the expression of odontogenic genes, including those encoding dentin sialophosphoprotein, dentin matrix protein 1, ALP, osteocalcin, and runt-related transcription factor 2. Moreover, Mg-BG substantially suppressed the secretion of inflammatory cytokines (interleukin [IL]-4, IL-6, IL-8, and tumor necrosis factor-alpha). Collectively, the results of this study suggest that Mg-BG has excellent in vitro bioactivity and is a potential material for vital pulp therapy of inflamed pulps.
Topics: Humans; Dental Pulp; Magnesium; Anti-Inflammatory Agents; Stem Cells; Glass; Odontogenesis; Cytokines; Cells, Cultured; Biocompatible Materials; X-Ray Diffraction; Spectroscopy, Fourier Transform Infrared; Alkaline Phosphatase; Ceramics; Materials Testing; Powders; Microscopy, Electron, Scanning
PubMed: 38740053
DOI: 10.1088/1748-605X/ad4ada -
PloS One 2024Flowable resin composites (FRC) are tooth-colored restorative materials that contain a lower filler particle content, and lower viscosity than their bulk counterparts,... (Comparative Study)
Comparative Study
BACKGROUND
Flowable resin composites (FRC) are tooth-colored restorative materials that contain a lower filler particle content, and lower viscosity than their bulk counterparts, making them useful for specific clinical applications. Yet, their chemical makeup may impact the cellular population of the tooth pulp. This in-vitro study assessed the cytocompatibility and odontogenic differentiation capacity of dental pulp stem cells (DPSCs) in response to two recent FRC material extracts.
METHODS
Extracts of the FRC Aura easyflow (AEF) and Polofil NHT Flow (PNF) were applied to DPSCs isolated from extracted human teeth. Cell viability of DPSCs was assessed using MTT assay on days 1, 3 and 7. Cell migration was assessed using the wound healing assay. DPSCs' capacity for osteo/odontogenic differentiation was assessed by measuring the degree of mineralization by Alizarin Red S staining, alkaline phosphatase enzyme (ALP) activity, and monitoring the expression of osteoprotegerin (OPG), RUNX Family Transcription Factor 2 (RUNX2), and the odontogenic marker dentin sialophosphoprotein (DSPP) by RT-PCR. Monomer release from the FRC was also assessed by High-performance liquid chromatography analysis (HPLC).
RESULTS
DPSCs exposed to PNF extracts showed significantly higher cell viability, faster wound closure, and superior odontogenic differentiation. This was apparent through Alizarin Red staining of calcified nodules, elevated alkaline phosphatase activity, and increased expression of osteo/odontogenic markers. Moreover, HPLC analysis revealed a higher release of TEDGMA, UDMA, and BISGMA from AEF.
CONCLUSIONS
PNF showed better cytocompatibility and enhancement of odontogenic differentiation than AEF.
Topics: Dental Pulp; Humans; Stem Cells; Cell Differentiation; Composite Resins; Cell Survival; Odontogenesis; Cell Movement; Cells, Cultured
PubMed: 38739591
DOI: 10.1371/journal.pone.0303154 -
Orthodontics & Craniofacial Research May 2024To explore the association between third molar agenesis and supernumerary tooth formation in a white-European population.
OBJECTIVES
To explore the association between third molar agenesis and supernumerary tooth formation in a white-European population.
MATERIALS AND METHODS
A record review in various orthodontic clinics identified 380 eligible white-European individuals, half of whom had non-syndromic permanent supernumerary teeth (122 males and 68 females, totalling 244 supernumerary teeth; median age: 13.1, iqr: 1.5 years), and the other half were age- and sex-matched controls with full dentition, excluding the third molars. Tooth sequences were identified in panoramic radiographs.
RESULTS
In the supernumerary group, approximately 80% of the individuals had a single supernumerary tooth, followed by those having two additional teeth. In both groups, there was no sexual dimorphism in third molar agenesis severity. The prevalence of third molar agenesis in the supernumerary group was similar to that of the control group (28/190 = 14.7% in both groups; p = 1.0). In total, 53 third molars were missing in the supernumerary group (n = 190) compared to 67 in the control group (n = 190; p = .862). The ratio of bilateral to unilateral third molar agenesis was significantly lower in the supernumerary group than in the control group (1.0 vs. 3.7, respectively; p = .026).
CONCLUSION
The presence of supernumerary teeth did not significantly alter the likelihood of third molar agenesis or its severity. Bilateral third molar agenesis was considerably less prevalent in individuals with supernumerary teeth compared to controls. The present novel findings have important clinical and developmental implications.
PubMed: 38721988
DOI: 10.1111/ocr.12807 -
Journal of Dentistry Jul 2024Three-dimensional (3D) cell culture systems perform better in resembling tissue or organism structures compared with traditional 2D models. Organs-on-chips (OoCs) are...
AIM
Three-dimensional (3D) cell culture systems perform better in resembling tissue or organism structures compared with traditional 2D models. Organs-on-chips (OoCs) are becoming more efficient 3D models. This study aimed to create a novel simplified dentin-on-a-chip using microfluidic chip technology and tissue engineering for screening dental materials.
METHODOLOGY
A microfluidic device with three channels was designed for creating 3D dental tissue constructs using stem cells from the apical papilla (SCAP) and gelatin methacrylate (GelMA). The study investigated the effect of varying cell densities and GelMA concentrations on the layer features formed within the microfluidic chip. Cell viability and distribution were evaluated through live/dead staining and nuclei/F-actin staining. The osteo/odontogenic potential was assessed through ALP staining and Alizarin red staining. The impact of GelMA concentrations (5 %, 10 %) on the osteo/odontogenic differentiation trajectory of SCAP was also studied.
RESULTS
The 3D tissue constructs maintained high viability and favorable spreading within the microfluidic chip for 3-7 days. A cell seeding density of 2 × 10 cells/μL was found to be the most optimal choice, ensuring favorable cell proliferation and even distribution. GelMA concentrations of 5 % and 10 % proved to be most effective for promoting cell growth and uniform distribution. Within the 5 % GelMA group, SCAP demonstrated higher osteo/odontogenic differentiation than that in the 10 % GelMA group.
CONCLUSION
In 3D culture, GelMA concentration was found to regulate the osteo/odontogenic differentiation of SCAP. The study recommends a seeding density of 2 × 10 cells/μL of SCAP within 5 % GelMA for constructing simplified dentin-on-a-chip.
CLINICAL SIGNIFICANCE
This study built up the 3D culture protocol, and induced odontogenic differentiation of SCAP, thus forming the simplified dentin-on-a-chip and paving the way to be used as a well-defined biological model for regenerative endodontics. It may serve as a potential testing platform for cell differentiation.
Topics: Tissue Engineering; Humans; Cell Differentiation; Cell Survival; Cell Proliferation; Lab-On-A-Chip Devices; Dentin; Gelatin; Dental Papilla; Stem Cells; Odontogenesis; Osteogenesis; Methacrylates; Cell Culture Techniques; Microfluidics; Cell Culture Techniques, Three Dimensional; Cells, Cultured
PubMed: 38719135
DOI: 10.1016/j.jdent.2024.105028 -
Acta Biomaterialia Jun 2024Dental pulp is the only soft tissue in the tooth which plays a crucial role in maintaining intrinsic multi-functional behaviors of the dentin-pulp complex. Nevertheless,...
Dental pulp is the only soft tissue in the tooth which plays a crucial role in maintaining intrinsic multi-functional behaviors of the dentin-pulp complex. Nevertheless, the restoration of fully functional pulps after pulpitis or pulp necrosis, termed endodontic regeneration, remained a major challenge for decades. Therefore, a bioactive and in-situ injectable biomaterial is highly desired for tissue-engineered pulp regeneration. Herein, a decellularized matrix hydrogel derived from porcine dental pulps (pDDPM-G) was prepared and characterized through systematic comparison against the porcine decellularized nerve matrix hydrogel (pDNM-G). The pDDPM-G not only exhibited superior capabilities in facilitating multi-directional differentiation of dental pulp stem cells (DPSCs) during 3D culture, but also promoted regeneration of pulp-like tissues after DPSCs encapsulation and transplantation. Further comparative proteomic and transcriptome analyses revealed the differential compositions and potential mechanisms that endow the pDDPM-G with highly tissue-specific properties. Finally, it was realized that the abundant tenascin C (TNC) in pDDPM served as key factor responsible for the activation of Notch signaling cascades and promoted DPSCs odontoblastic differentiation. Overall, it is believed that pDDPM-G is a sort of multi-functional and tissue-specific hydrogel-based material that holds great promise in endodontic regeneration and clinical translation. STATEMENT OF SIGNIFICANCE: Functional hydrogel-based biomaterials are highly desirable for endodontic regeneration treatments. Decellularized extracellular matrix (dECM) preserves most extracellular matrix components of its native tissue, exhibiting unique advantages in promoting tissue regeneration and functional restoration. In this study, we prepared a porcine dental pulp-derived dECM hydrogel (pDDPM-G), which exhibited superior performance in promoting odontogenesis, angiogenesis, and neurogenesis of the regenerating pulp-like tissue, further showed its tissue-specificity compared to the peripheral nerve-derived dECM hydrogel. In-depth proteomic and transcriptomic analyses revealed that the activation of tenascin C-Notch axis played an important role in facilitating odontogenic regeneration. This biomaterial-based study validated the great potential of the dental pulp-specific pDDPM-G for clinical applications, and provides a springboard for research strategies in ECM-related regenerative medicine.
Topics: Dental Pulp; Animals; Hydrogels; Swine; Regeneration; Stem Cells; Decellularized Extracellular Matrix; Cell Differentiation; Regenerative Endodontics; Humans; Tissue Engineering
PubMed: 38692468
DOI: 10.1016/j.actbio.2024.04.040 -
Dentistry Journal Mar 2024Dental development defects (DDDs) are quantitative and/or qualitative alterations produced during odontogenesis that affect both primary and permanent dentition. The...
Dental development defects (DDDs) are quantitative and/or qualitative alterations produced during odontogenesis that affect both primary and permanent dentition. The etiology remains unknown, being associated with prenatal, perinatal, and postnatal factors. The aims were to identify the possible etiological factors, as well as the prevalence of DDDs in the primary and permanent dentition in a pediatric population. Two hundred twenty-one children between 2 and 15 years of age, patients of the master's degree in Pediatric Dentistry of the Complutense University of Madrid, were reviewed. DDDs were observed in 60 children. Next, a cross-sectional, case-control study was carried out (60 children in the control group and 60 children in the case group). The parents or guardians completed a questionnaire aimed at identifying associated etiological factors. The prevalence of DDDs in patients attending our master's program in both dentitions was 27.15%. Otitis, tonsillitis, high fevers, and medication intake stood out as the most relevant postnatal factors among cases and controls. The permanent maxillary right permanent central incisor and the primary mandibular right second molar were the most affected; there were no differences in relation to gender. One out of three children who presented DDDs in the primary dentition also presented DDDs in the permanent dentition. Prenatal and postnatal etiological factors showed a significant relationship with DDD alterations, considered risk factors for DDDs in both dentitions.
PubMed: 38667996
DOI: 10.3390/dj12040084 -
Journal of Functional Biomaterials Apr 2024This study investigated the incorporation of sources of calcium, phosphate, or both into electrospun scaffolds and evaluated their bioactivity on human dental pulp cells...
This study investigated the incorporation of sources of calcium, phosphate, or both into electrospun scaffolds and evaluated their bioactivity on human dental pulp cells (HDPCs). Additionally, scaffolds incorporated with calcium hydroxide (CH) were characterized for degradation, calcium release, and odontogenic differentiation by HDPCs. Polycaprolactone (PCL) was electrospun with or without 0.5% / of calcium hydroxide (PCL + CH), nano-hydroxyapatite (PCL + nHA), or β-glycerophosphate (PCL + βGP). SEM/EDS analysis confirmed fibrillar morphology and particle incorporation. HDPCs were cultured on the scaffolds to assess cell viability, adhesion, spreading, and mineralized matrix formation. PCL + CH was also evaluated for gene expression of odontogenic markers (RT-qPCR). Data were submitted to ANOVA and Student's -test (α = 5%). Added CH increased fiber diameter and interfibrillar spacing, whereas βGP decreased both. PCL + CH and PCL + nHA improved HDPC viability, adhesion, and proliferation. Mineralization was increased eightfold with PCL + CH. Scaffolds containing CH gradually degraded over six months, with calcium release within the first 140 days. CH incorporation upregulated and expression after 7 and 14 days. In conclusion, CH- and nHA-laden PCL fiber scaffolds were cytocompatible and promoted HDPC adhesion, proliferation, and mineralized matrix deposition. PCL + CH scaffolds exhibit a slow degradation profile, providing sustained calcium release and stimulating HDPCs to upregulate odontogenesis marker genes.
PubMed: 38667554
DOI: 10.3390/jfb15040097 -
Biomedical Materials (Bristol, England) May 2024The decellularized matrix has a great potential for tissue remodeling and regeneration; however, decellularization could induce host immune rejection due to incomplete...
The decellularized matrix has a great potential for tissue remodeling and regeneration; however, decellularization could induce host immune rejection due to incomplete cell removal or detergent residues, thereby posing significant challenges for its clinical application. Therefore, the selection of an appropriate detergent concentration, further optimization of tissue decellularization technique, increased of biosafety in decellularized tissues, and reduction of tissue damage during the decellularization procedures are pivotal issues that need to be investigated. In this study, we tested several conditions and determined that 0.1% Sodium dodecyl sulfate and three decellularization cycles were the optimal conditions for decellularization of pulp tissue. Decellularization efficiency was calculated and the preparation protocol for dental pulp decellularization matrix (DPDM) was further optimized. To characterize the optimized DPDM, the microstructure, odontogenesis-related protein and fiber content were evaluated. Our results showed that the properties of optimized DPDM were superior to those of the non-optimized matrix. We also performed the 4D-Label-free quantitative proteomic analysis of DPDM and demonstrated the preservation of proteins from the natural pulp. This study provides a optimized protocol for the potential application of DPDM in pulp regeneration.
Topics: Dental Pulp; Proteomics; Animals; Tissue Engineering; Tissue Scaffolds; Decellularized Extracellular Matrix; Sodium Dodecyl Sulfate; Humans; Odontogenesis; Extracellular Matrix
PubMed: 38653259
DOI: 10.1088/1748-605X/ad4245 -
Cellular and Molecular Biology... Mar 2024This study aimed to evaluate the physiological role of NAMPT associated with MDPC-23 odontoblast cell proliferation. Cell viability was measured using the (DAPI)...
This study aimed to evaluate the physiological role of NAMPT associated with MDPC-23 odontoblast cell proliferation. Cell viability was measured using the (DAPI) staining, caspase activation analysis and immunoblotting were performed. Visfatin promoted MDPC-23 odontoblast cell growth in a dose-dependent manner. Furthermore, the up-regulation of Visfatin promoted odontogenic differentiation and accelerated mineralization through an increase in representative odontoblastic biomarkers in MDPC-23 cells. However, FK-866 cell growth in a dose-dependent manner induced nuclear condensation and fragmentation. FK-866-treated cells showed H&E staining and increased apoptosis compared to control cells. The expression of anti-apoptotic factors components of the mitochondria-dependent intrinsic apoptotic pathway significantly decreased following FK-866 treatment. The expression of pro-apoptotic increased upon FK-866 treatment. In addition, FK-866 activated caspase-3 and PARP to induce cell death. In addition, after treating FK-866 for 72 h, the 3/7 activity of MDPC-23 cells increased in a concentration-dependent manner, and the IHC results also confirmed that Caspase-3 increased in a concentration-dependent. Therefore, the presence or absence of NAMPT expression in dentin cells was closely related to cell proliferation and formation of extracellular substrates.
Topics: Nicotinamide Phosphoribosyltransferase; Apoptosis; Cell Proliferation; Odontoblasts; Animals; Mice; Cell Line; Cytokines; Caspase 3; Cell Differentiation; Cell Survival; Acrylamides; Odontogenesis
PubMed: 38650160
DOI: 10.14715/cmb/2024.70.3.4