Periodontal ligament (PDL) has become an elective source of mesenchymal stem cells (PDLSCs) in dentistry. This research aimed to compare healthy PDLSCs (hPDLSCs) and periodontitis PDLSCs (pPDLSCs) to ascertain any possible functional differences owing to their milieux of origin. Cells were tested in terms of colony-forming unit efficiency; multi differentiating capacity; immunophenotype, stemness, and senescent state were studied by flow cytometry, immunofluorescence, and β-galactosidase staining; gene expression using RT-PCR. Both hPDLSCs and pPDLSCs were comparable in terms of their immunophenotype and multilineage differentiation capabilities, but pPDLSCs showed a senescent phenotype more frequently. Thus, a selective small molecule inhibitor of DNA methyltransferase (DNMT), RG108, known for its effect on senescence, was used to possibly reverse this phenotype. RG108 did not affect the proliferation and apoptosis of PDLSCs, and it showed little effect on hPDLSCs, while a significant reduction of both p16 and p21 was detected along with an increase of SOX2 and OCT4 in pPDLSCs after treatment at 100 μM RG108. Moreover, the subset of PDLSCs co-expressing OCT4 and p21 decreased, and adipogenic potential increased in pPDLSCs after treatment. pPDLSCs displayed a senescent phenotype that could be reversed, opening new perspectives for the treatment of periodontitis.

The mechanical and biological behaviors of PMMA/Al₂O₃ composites incorporating 30 wt.%, 40 wt.%, and 50 wt.% of Al₂O₃ were thoroughly characterized as regards to their possible application in implant-supported prostheses. The Al₂O₃ particles accounted for an increase in the flexural modulus of PMMA. The highest value was recorded for the composite containing 40 wt.% Al₂O₃ (4.50 GPa), which was about 18% higher than that of its unfilled counterpart (3.86 GPa). The Al₂O₃ particles caused a decrease in the flexural strength of the composites, due to the presence of filler aggregates and voids, though it was still satisfactory for the intended application. The roughness (Ra) and water contact angle had the same trend, ranging from 1.94 µm and 77.2° for unfilled PMMA to 2.45 µm and 105.8° for the composite containing the highest alumina loading, respectively, hence influencing both the protein adsorption and cell adhesion. No cytotoxic effects were found, confirming that all the specimens are biocompatible and capable of sustaining cell growth and proliferation, without remarkable differences at 24 and 48 h. Finally, Al₂O₃ was able to cause strong cell responses (cell orientation), thus guiding the tissue formation in contact with the composite itself and not enhancing its osteoconductive properties, supporting the PMMA composite’s usage in the envisaged application.

The role of periodontal ligament stem cells (PDLSCs) and dental pulp stem cells (DPSCs) in stimulating angiogenesis has been reported, but their angiogenetic potential has not been directly compared. In this work, paired PDLSCs and DPSCs, i.e., derived from the same donor, were tested for their immunophenotype and multi-differentiation capabilities, with particular emphasis on their pro-angiogenic activity. Flow cytometry was utilized to study the expression of mesenchymal stem cell, pericyte, and endothelial markers, while gene expression was evaluated through real-time PCR. The angiogenic potential was assessed recurring to tubulogenesis assay, co-cultures with Human Microvascular Endothelial Cell (HMEC-1), and VEGF-A quantification. The immunophenotype of DPSCs and PDLSCs was different in CD146+ and CD31+ cell subsets, but both cell types promoted HMEC-1 tubulogenesis in vitro. Consistently, VEGF-A gene expression level and its quantification in cell-conditioned media of PDLSCs and DPSCs was comparable between them, and both promoted the formation of vessel-like structures, when co-cultured with HMEC-1 cells. All together, these results showed the heterogeneity of PDLSCs and DPSCs, which are constituted of different cellular subsets, likely modulated by the microenvironmental cues. PDLSCs and DPSCs showed comparable pro-angiogenic activity, enhanced by the contemporary expression of angiogenic and chemotactic factors.

This work focuses on the study and comparison of two mixing methods for the dispersion of Alumina-Toughened Zirconia (ATZ) within the polymer matrix of Poly(ε-caprolactone) (PCL). The dry-mixing method using solvent-free impact milling (M) and the solvent casting method with chloroform (SC) were investigated. Samples were produced by 3D printing, and specimens were printed at increasing ATZ loadings (namely, 10, 20, and 40 wt.%). The chemico-physical, mechanical, and cell interaction characteristics of the materials prepared with both mixing methods were studied. Solvent mixing allowed better dispersion of the ATZ in the polymer matrix with respect to dry mixing. In addition, dry mixing affected the molecular weight of the PCL/ATZ composites much more than the solvent casting method. For these reasons, materials obtained by solid mixing exhibited the worst mechanical performance with respect to those obtained by solvent casting, which showed increased Young’s moduli with increasing ATZ amounts. The in vitro biological response elicited in a mesenchymal stem cell model seemed to be influenced by the mixing method, with a preference for the composites obtained through solvent mixing and containing 20 or 40 wt.% of ATZ.

To evaluate the fracture strength and linear elongation at break of three-units fixed partial dentures (FPDs) fabricated with traditional and new materials for fixed prosthodontics before and after ageing. Sixty models of three-units FPDs were fabricated and cemented onto a Co-Cr model simulating the replacement of a maxillary second premolar. The samples were randomly divided into 3 groups: metal-ceramic (MCR), graphene-doped polymethylmethacrylate (PMMA-GR) and polymethylmethacrylate (PMMA). Half of the samples were directly subjected to fracture test, while the remaining half underwent an ageing process and then a fracture loading test using an electrodynamic testing machine. Fracture load and elongation at break values were taken and statistically analysed. Significant differences were detected between the different materials (p<0.05). All groups showed a reduction of the fracture load and elongation at break values after ageing, but not statistically significant, except for PMMA group (p = 2.012e-19) (p = 3.8e-11). MCR and PMMA-GR three-units FPDs showed higher fracture strength and lower elongation at break compared to PMMA. MCR and PMMA-GR had higher resistance to ageing processes compared to PMMA. PMMA-GR could be considered a material for long-term provisional restorations as its mechanical behaviour and ageing resistance are more like MCR than PMMA.

The increasing demand for smart bone substitutes has boosted the implementation of biomaterials possibly endowed with both pro-osteogenic and pro-angiogenic capabilities, among which bioactive glasses hold great potential. Hence, two Poly(ε-caprolactone) (PCL)-based composites were loaded at 10 wt.%, with either pristine (SBA3) or copper-doped (SBA3_Cu) silica-based bioactive glasses, through a solvent casting method with chloroform. Neat PCL was used as a control. Samples produced by 3D printing underwent SEM and EDX analyses, and the following were measured: tensile strength and hardness, surface roughness, ion release through ICP-OES, surface free energy, and optical contact angle. Adipose-derived mesenchymal stem cells (ASCs) and human microvascular endothelial cells (HMEC-1) were used to test the biocompatibility of the materials through cell adhesion, spreading, and viability assays. A significant improvement in tensile strength and hardness was observed especially with Cu-doped composites. Both SBA3 and SBA3_Cu added to the PCL favored the early adhesion and the proliferation of HMEC-1 after 3 and 7 days, while ASCs proliferated significantly the most on the SBA-containing composite, at all the time points. Cellular morphology analysis highlighted interesting adaptation patterns to the samples. Further biological characterizations are needed to understand thoroughly how specific bioactive glasses may interact with different cellular types.