Polishing methods for composites restoration: the influence on human gingival fibroblasts behaviour.

BACKGROUND: Carious/Non-carious cervical lesions with gingival recessions may require both dental and periodontal reconstructive therapy, where flaps/grafts may be placed in contact with a dental filling material. Human Gingival Fibroblasts (HGF-1) response during the early phase of healing could vary according to the procedures employed to cure the dental composite. Moreover, oxygen diffusion into dental composite inhibits the polymerization reaction, creating an oxygen-inhibited layer (OIL) that presents residual unreacted monomers. The aim of this study was to assess the effect of different polishing techniques and OIL on HGF-1. METHODS: Composite discs polished with different techniques (diamond rubber, abrasive discs and tungsten carbide burr) were used. An additional not polished smooth group obtained with and without OIL was used as control. Samples were physically characterized through the analysis of their hydrophilicity and surface topography through contact angle measurement and SEM, respectively; afterwards the biologic response of HGF-1 when cultured on the different substrates was analyzed in terms of cytotoxicity and gene expression. RESULTS: The finishing systems caused alterations to the wettability, even if without a proportional relation towards the results of the proliferation essay, from which emerges a greater proliferation on surfaces polished with one-step diamond rubber and with abrasive discs as well as a direct effect of the glycerin layer, confirming that surface roughness can heavily influence the biological response of HGF-1. CONCLUSIONS: Surfaces wettability as well as cellular behavior seem to be affected by the selection of the finishing system used to lastly shape the restoration. Especially, the presence of OIL act as a negative factor in the regards of human gingival fibroblasts. The present study may provide the first clinical instruction regarding the best polishing system of composite material when the restoration is placed directly in contact with soft tissue cells. Understanding HGF-1 behavior can help identifying the polishing treatment for direct restoration of carious/non-carious cervical lesions associated with gingival recessions.

Predictability of intraoral scanner error for full-arch implant-supported rehabilitation.

OBJECTIVES: The present study aimed to analyze the behaviors of three intraoral scanners (IOSs): evaluating the interdistance and axial inclination discrepancies in full-arch scans, predictable errors were searched. MATERIALS AND METHODS: Six edentulous sample models with variable numbers of dental implants were used; reference data were obtained with a coordinate-measuring machine (CMM). Each IOS (i.e., Primescan, CS3600, and Trios3) performed 10 scans per model (180 total scans). The origin of each scan body was used as a reference point to measure interdistance lengths and axial inclinations. Precision and trueness of interdistance measurements and axial inclinations were evaluated to address error predictability. Bland-Altman analysis, followed by linear regression analysis and Friedman's test (plus Dunn's post hoc correction), was performed to evaluate the precision and trueness. RESULTS: Regarding interdistance, Primescan showed the best precision (mean ± SD: 0.047 ± 0.020 mm), while Trios3 underestimated the reference value more than the others (p < 0.001) and had the worst performance (mean ± SD: -0.079 ± 0.048 mm). Concerning the inclination angle, Primescan and Trios3 tended to overestimate angle values, while CS3600 underestimated them. Primescan had fewer inclination angle outliers, but it tended to add 0.4-0.6° to the measurements. CONCLUSIONS: IOSs showed predictable errors: they tended to overestimate or underestimate linear measurements and axial inclinations of scan bodies, one added 0.4-0.6° to the angle inclination values. In particular, they showed heteroscedasticity, a behavior probably related to the software or the device itself. CLINICAL SIGNIFICANCE: IOSs showed predictable errors that could affect clinical success. When performing a scan or choosing a scanner, clinicians should clearly know their behaviors.

Photogrammetric method to measure the discrepancy between clinical and software-designed positions of implants.

STATEMENT OF PROBLEM: The position of dental implants placed with software-guided systems should be highly accurate in order to ensure safety and a passive fit of the immediate prosthesis. PURPOSE: The purpose of this study was to measure the discrepancy between the clinical and software-planned position of dental implants by applying a photogrammetric method. MATERIAL AND METHODS: Two casts were obtained, 1 from the surgical template and 1 from the actual position of the implants on the alveolar ridge of a patient. Photogrammetry was then applied to precisely locate the position of each implant on the casts. Because this mathematical technique required the identification of image points and of the relative spatial coordinates, 4 marks were drilled on the implant screw. The position of the implants was then identified as the geometric center of the 4 marks, while the orientation of the implant axis was represented by a vector normal to the plane fitting the points. A series of 16 convergent images all around the object was made using a high-resolution digital camera. A mathematical method called "rototranslation" was used to superimpose the cast images for the comparison. RESULTS: The tests performed on the casts resulted in an average precision level of 4 µm for the locations and less than 1 degree for the axis of the implants. A series of empirical and numerical tests were performed to assess the performance of the procedure and of the measurement protocol. CONCLUSIONS: The photogrammetric method is reproducible and can be used to measure the discrepancy between the software-planned and the real position of dental implants. Considering that the average precision level required for an implant-based prosthesis is approximately 50 µm, the error associated with this method can be considered as negligible.

The top 100 most-cited articles in dentistry by authors with Italian affiliation.

INTRODUCTION: Analyzing highly-cited articles can provide a retrospective assessment of research evolution and predict future developments. The aim of this study was to carry out a bibliometric analysis of the 100 most-cited articles in dentistry by authors with an Italian affiliation. EVIDENCE ACQUISITION: All the articles published in journals indexed under the Scopus category "Dentistry" and with at least one author affiliated to any Italian institution were searched in September 2022. The 100 most-cited articles were selected and relevant data were extracted and summarized. The analysis of co-authorship at country level and co-occurrence of keywords was carried out. EVIDENCE SYNTHESIS: The 100 most-cited articles were published between 1976 and 2020. The number of citations ranged from 235 to 1683, with a mean of 361. The series included 25 citation classics (>400 citations). The majority of articles were classified as expert opinion/narrative reviews (N.=47). Almost half of the articles refer to three predominant disciplines which were implantology, periodontology and restorative dentistry. Only two articles were single-authored. In 30 articles, all the authors had an Italian affiliation and the US was the most frequent country for non-Italian authors. Only one article was published in journals owned by a non-Italian publisher. CONCLUSIONS: The present series of highly-cited articles confirms the important role of Italy in dental research. We found an absence of correlation between the level of evidence and the number of citations and a non-homogeneous distribution of highly-cited papers in the different dental disciplines. The majority of articles included in the series shared international co-authorship and were published in high-impact journals.

Relationship between periodontal disease and obstructive sleep apnea in adults: A systematic review.

The purpose of this systematic review was to evaluate whether there are scientific evidence regarding the association between periodontitis and obstructive sleep apnea (OSA) in adults. An electronic search was performed on MEDLINE/PubMed for prospective and retrospective longitudinal studies, cohort studies, and case-control studies conducted in human adults affected by both OSA and periodontitis. Two reviewers extracted the data using a custom Excel spreadsheet. A methodological assessment of the quality of the studies was performed using the Newcastle-Ottawa Scale. Fourteen studies were included. All studies evaluated the association between periodontitis and OSA. None of the studies evaluated the cause-effect relationship. Eleven studies found a significant positive relationship between periodontitis and OSA, whereas three found no statistically significant association. Several study limitations were observed, such as lack of standardization of study groups, diagnosis of periodontitis and OSA, and differences in study design. Evidence of a plausible association between periodontitis and OSA was found. The possible relationship could be explained by systemic inflammation, oral breathing, and the comorbid relationship attributable to common risk factors. Observational and randomized controlled studies are needed to clarify the mechanism of interaction between the two conditions.

Aptamer-enriched scaffolds for tissue regeneration: a systematic review of the literature.

Introduction: Aptamers are a brand-new class of receptors that can be exploited to improve the bioactivity of tissue engineering grafts. The aim of this work was to revise the current literature on in vitro and in vivo studies in order to i) identify current strategies adopted to improve scaffold bioactivity by aptamers; ii) assess effects of aptamer functionalization on cell behavior and iii) on tissue regeneration. Methods: Using a systematic search approach original research articles published up to 30 April 2022, were considered and screened. Results: In total, 131 records were identified and 18 were included in the final analysis. Included studies showed that aptamers can improve the bioactivity of biomaterials by specific adsorption of adhesive molecules or growth factors from the surrounding environment, or by capturing specific cell types. All the studies showed that aptamers ameliorate scaffold colonization by cells without modifying the physicochemical characteristics of the bare scaffold. Additionally, aptamers seem to promote the early stages of tissue healing and to promote anatomical and functional regeneration. Discussion: Although a metanalysis could not be performed due to the limited number of studies, we believe these findings provide solid evidence supporting the use of aptamers as a suitable modification to improve the bioactivity of tissue engineering constructs.

How Is Endodontics Taught in Italy? A Survey of Italian Dental Schools.

The aim of our study was to investigate how endodontics is taught in Italian universities. An online survey was conducted from August to December 2021. A comparison between courses led by full or associate professors (Group 1) versus courses led by other figures, such as researchers or temporary lecturers (Group 2), was made. A total of 28 out of 36 schools participated (78%). In most schools, endodontics is taught in the fifth year to 15-29 students. All schools planned pre-clinical endodontic training, and in 25/28 schools (89.3%), clinical endodontic training was also provided. The course programs varied among schools, and significantly more hours were allocated to teaching nonsurgical root canal treatment in Group 1 schools than in Group 2 schools. The average numbers of hours of preclinical and clinical training were 34.3 ± 23.6 and 84.1 ± 76.7, respectively. All schools used rotary NiTi files in their clinical training, and the vertical condensation of hot gutta-percha was the most frequently taught obturation technique. As expected, the scenario of endodontic education in Italian universities was variable and needs harmonization. Courses led by full or associate professors seem to be better structured.

Oral status of a noble European couple from the 16th century: A morphologic analysis of the teeth of Alessandro Farnese and his wife Maria D'Aviz.

Objective. To report the results of dental morphological analysis performed on the remains of two European nobles (Duke Alessandro Farnese and his wife Maria D'Aviz) who lived in the 16th century, together with hypotheses on their nutrition and oral hygiene habits. Design and results. The remains of Alessandro Farnese (1545-1592) and Princess Maria D'Aviz (1538-1577) were exhumed in Parma, Italy, in the context of an historical investigation into their possible causes of death. The skulls were examined and analysed through direct inspection, high-detailed photographs and radiographs. Ante mortem tooth loss (AMTL), postmortem tooth loss (PMTL), Scott and Smith dental wear indexes and the Kerr periodontal index were used to assess the dental and periodontal status of the couple. Alessandro Farnese suffered from severe dental wear while it was presumed that Maria D'Aviz was affected by periodontal disease and tooth decay. Conclusions. Based on the findings of the present analysis, we hypothesise that Alessandro Farnese's diet was mainly based on hard and unrefined foods, also suggested by limited historical reports. It is likely that Maria D'Aviz's nutrition was based on sugar-enriched foods.

Preparation of hybrid samples for scanning electron microscopy (SEM) coupled to focused ion beam (FIB) analysis: A new way to study cell adhesion to titanium implant surfaces.

The study of the intimate connection occurring at the interface between cells and titanium implant surfaces is a major challenge for dental materials scientists. Indeed, several imaging techniques have been developed and optimized in the last decades, but an optimal method has not been described yet. The combination of the scanning electron microscopy (SEM) with a focused ion beam (FIB), represents a pioneering and interesting tool to allow the investigation of the relationship occurring at the interface between cells and biomaterials, including titanium. However, major caveats concerning the nature of the biological structures, which are not conductive materials, and the physico-chemical properties of titanium (i.e. color, surface topography), require a fine and accurate preparation of the sample before its imaging. Hence, the aim of the present work is to provide a suitable protocol for cell-titanium sample preparation before imaging by SEM-FIB. The concepts presented in this paper are also transferrable to other fields of biomaterials research.

Testosterone Enanthate: An In Vitro Study of the Effects Triggered in MG-63 Cells.

The aim of this study was to investigate the effects of the androgenic hormone testosterone enanthate (TE) on human MG-63 cells. MG-63 were cultured for 24 h in the presence of TE at increasing concentrations to assess its lethal dose. Therefore, the suitable concentration for a prolonged use of TE in vitro was assessed by viability assay over 9 days. Finally, MG-63 were exposed to TE for 14 days and assayed for differentiation by qPCR and Alizarin Red S staining. TE in the amount of 100 µM resulted as the maximum dose tolerated by MG-63 cells after 24 h. However, a prolonged exposure in culture TE in the amount of 100 µM showed a cytostatic effect on cell proliferation. On the contrary, TE 10 µM was tolerated by the cells and did not boost cell proliferation, but did enhance new bone formation, as revealed by COL1A1, ALPL, BGLAP, and IBSP gene expression after 3, 7, and 14 days, and calcium deposition by Alizarin Red S staining after 14 days. Based on the current study, 10 µM is the critical dose of TE that should be used in vitro to support bone differentiation of MG-63 cells.

The effect of a 2-mm inter-implant distance on esthetic outcomes in immediately non-occlusally loaded platform shifted implants in healed ridges: 12-month results of a randomized clinical trial.

BACKGROUND: Three millimeter is considered as the minimum distance to obtain soft and bone tissue stability in case of adjacent implants. The possibility to preserve peri-implant bone level using a platform switching connection has questioned this concept. PURPOSE: The study evaluates soft tissue maintenance and marginal bone stability around implants, placed at 2 or 3 mm of distance. MATERIALS AND METHODS: Thirty patients received two immediately loaded implants either at 2-mm (test) or at 3-mm (control) of distance in the premolar area. Soft tissue esthetics (papilla height and fill, keratinized tissue, recession) and radiographic peri-implant bone level changes were measured at 3, 6, and 12 months. RESULTS: No significant differences between the two groups were detected neither for all soft tissue esthetic outcomes nor for bone level modifications up to 12 months. CONCLUSION: The results suggested that up to 12 months post-loading, both 2- and 3-mm inter-distance platform-switched implants in healed site, supported adequate esthetic outcomes and peri-implant bone stability.

A glance on the role of fibronectin in controlling cell response at biomaterial interface.

The bioactivity of biomaterials is closely related to cell response in contact with them. However, shortly after their insertion, materials are soon covered with proteins that constitute the biological fluids, and which render the direct surface recognition by cells almost impossible. The control of protein adsorption at the interface is therefore desirable. Extracellular matrix proteins are of particular interest in this sense, due to their well-known ability to modulate cell behavior. Particularly, fibronectin plays a leading role, being present in both healthy and injured tissues undergoing healing and regeneration. The aim of the present work is to give an overview on fibronectin and on its involvement in the control of cell behavior providing evidence of its pivotal role in the control of cell adhesion, spreading, migration, proliferation and differentiation. A deep insight into methods to enrich biomaterials surface with fibronectin will be then discussed, as well as new cues on the possibility to design tailored platforms able to specifically retain fibronectin from the surrounding extracellular milieu.

Thermal treatment to increase titanium wettability induces selective proteins adsorption from blood serum thus affecting osteoblasts adhesion.

OBJECTIVES: To investigate how a thermal treatment to increase titanium wettability influences proteins adsorption from blood serum and osteoblasts responses. METHODS: Titanium discs with machined or micro-rough profiles were thermally treated to obtain hydrophilic surfaces. The adsorption kinetics of two representative serum proteins were determined by Bradford assay, while the stable protein adsorption pattern from blood serum was investigated by SDS-PAGE and Western Blot analysis. Subsequently, MC3T3-E1 cells were cultured on titanium for 24h and assayed for adhesion and morphology. RESULTS: Thermally-induced hydrophilicity dramatically improved the capacity of titanium to selectively adsorb fibronectin and fibrinogen from blood serum, without evident influence on other representative serum proteins. The selective adsorption of fibronectin was linked to the improved capacity of MC3T3-E1 cells to adhere and spread on hydrophilic surfaces. SIGNIFICANCE: We identified a potential method to improve selective protein adsorption on titanium by enhancing implant surface wettability through a thermal treatment. Selective fibronectin adsorption was further indicated as the responsible for improved osteoblasts adhesion. Targeting specific cell response by selective protein adsorption appears to be crucial to conceive even more performant therapies.

Titanium dental implants hydrophilicity promotes preferential serum fibronectin over albumin competitive adsorption modulating early cell response.

In vitro studies have consistently shown that titanium surface wettability affects the response of osteoprogenitors, leading to important advances in the clinical osseointegration of dental implants. However, the underlying molecular mechanisms remain unknown. Since surface conditioning by blood components initiates within milliseconds after insertion, it is reasonable to hypothesize that the amount and the type of blood proteins adsorbed influences the interaction between the implant surface and osteoprogenitors. To test this hypothesis, titanium implant surfaces with different characteristics, in terms of topography and wettability, have been conditioned with selected plasma proteins. Pure fibronectin (HFN) and albumin (HSA) solutions, or their mixture at the relative plasma concentrations were allowed to adsorb on titanium surfaces for 60 min. Protein adsorption was monitored by Bradford assay, while the contribution of HSA and HFN in forming the microfilm layer at the interface was studied by Western Blot. Subsequently, the same protein-conditioned surfaces were used to culture C2C12 cells, thus studying their capacity to adhere and to spread after 3 h. Cell viability was evaluated up to 7 days, while the expression of osteogenic genes was assessed after 3 days. Under competitive adsorption conditions, hydrophilicity promotes the selectivity of titanium for HFN regardless of the surface microtopography. As a consequence of selective HFN adsorption, cells on hydrophilic surfaces displayed enhanced adhesion and spreading, as well as increased proliferation. On the other hand, selective HFN adsorption did not appreciably affect cell differentiation. These data suggest that implant surface hydrophilicity plays a key role in guiding the selective adsorption of specific proteins from blood plasma. Moreover, the selective adsorption of HFN, as a consequence of surface hydrophilicity, was found to account for early cell responses amelioration. Thus, titanium surface hydrophilicity contributes to the clinical success of dental implant by selectively controlling protein adsorption at the interface.

Tetracalcium phosphate and biphasic tetracalcium phosphate/tricalcium phosphate powders' effects evaluation on human osteoblasts.

BACKGROUND: Calcium ions levels in bone niches have been demonstrated to severely influence new bone formation. Osteoinductive scaffolds containing calcium have been largely studied to control the release of calcium in bone regeneration and tissue engineering purpose. The aim of the present study was, firstly, to synthesize two different resorbable calcium phosphate-based powders, thought to be reservoirs of calcium ions, and secondary, to investigate their effects on human osteoblasts, in order to develop a suitable titanium coating material. METHODS: Tetracalcium phosphate (A450) and biphasic tetracalcium phosphatae/tricalcium phosphate (A850) powders were prepared with an innovative method. The presence of calcium phosphate structures was chemically confirmed with XRD. Furthermore, powders macroscopic aspect was observed with a stereomicroscope. For in-vitro experiments, human osteoblastic cells were cultured in the presence of A450 and A850, and assayed for viability and metabolic activity through Crystal Violet and MTT, respectively. RESULTS: Our synthesis led to the formation of calcium phosphates in both samples, even though A850 presented a higher level of crystallinity and a more powdery aspects than A450. Both the samples enhanced the viability of cultured cells, inhibiting cell metabolic activity in the case of A850, which furthermore showed to be internalized by cells. CONCLUSIONS: We developed two different kind of calcium phosphate-based powders and we tested their effect on human osteoblasts, underlying the possibility of use calcium phosphate-based coatings to enhance cell response on implantable materials.

Thermal-induced hydrophilicity enhancement of titanium dental implant surfaces.

Titanium surface characteristics, including microtopography, chemical composition, and wettability, are essential features to achieve osseointegration of dental implants, but the choice of a particular surface topography is still a debated topic among clinicians. An increased level of implant surface hydrophilicity has been demonstrated to ameliorate osseointegration and shorten healing times. The aim of this work is to develop and test a suitable thermal-based method to enhance titanium surface wettability without modifying other characteristics of the implant surface. For this function, titanium discs with different surface topography have been thermally treated by testing different temperatures and excluding those that led to evident chromatic and morphological modifications. The selected surface gain in wettability after the treatment was assessed through contact angle measurement, chemistry modifications through x-ray photoelectron spectroscopy (XPS) analysis, and microtopography through scanning electron microscopy (SEM). Results showed a great enhancement in hydrophilicity on the tested surfaces without any other modification in terms of surface chemical composition and topography. A possible limitation of this method could be the persistent, although relatively slow, biological aging of the surfaces after the treatment. The present findings indicate that the described treatment could be a safe and effective method to enhance dental titanium hydrophilicity and thus its biological performance.

Is selective protein adsorption on biomaterials a viable option to promote periodontal regeneration?

Periodontitis is an inflammatory condition that can induce significant destruction of the periodontium, the set of specialized tissues that provide nourishment and support to the teeth. According to the guided tissue regeneration principles, the periodontium can be regenerated if the spatiotemporal control of wound healing is obtained, namely the tune control of cell response. After material implantation, protein adsorption at the interface is the first occurring biological event, which influences subsequent cell response. With the regard of this, we hypothesize that the control of selective adsorption of biological cues from the surrounding milieu may be a key-point to control selective cell colonization of scaffolds for periodontal tissue regeneration.

Plasma Proteins at the Interface of Dental Implants Modulate Osteoblasts Focal Adhesions Expression and Cytoskeleton Organization.

The host-material interface is a crucial relationship dictating the possibility of successful osseointegration in implant dentistry. The aim of the present study was to characterize the effects of plasma proteins pre-adsorption on the adhesion capacity of osteoblasts, which occurs immediately after implant insertion in vivo. After having pre-adsorbed human plasma proteins on a machined and microrough titanium surface, MC3T3-E1 osteoblasts adhesion was evaluated through crystal violet cell adhesion assay, immunofluorescence staining for cytoskeleton, focal adhesions and cell nuclei, and scanning electron microscopy. The pre-adsorbed protein layer markedly affected the adhesion rate of cells, as well as their morphology and the expression of focal contacts. Moreover, protein adsorption to the underlying titanium surface was found to be correlated to surface pre-wetting. Thus, the early adsorption of serum proteins to the interface of dental implants impacts cell adhesion in terms of strength and of focal adhesions expression.

Functional Fibronectin Adsorption on Aptamer-Doped Chitosan Modulates Cell Morphology by Integrin-Mediated Pathway.

A decisive step in cell-biomaterial interaction is represented by the adsorption of proteins at the interface, whose fine control may be useful to trigger proper cell response. To this purpose, we can selectively control protein adsorption on biomaterials by means of aptamers. Aptamers selected to recognize fibronectin dramatically enhance chitosan ability to promote cell proliferation and adhesion, but the underlying biological mechanism remains unknown. We supposed that aptamers contributed to ameliorate the adsorption of fibronectin in an advantageous geometrical conformation for cells, thus regulating their morphology by the proper activation of the integrin-mediated pathway. We investigated this possibility by culturing epithelial cells on chitosan enriched with increasing doses of aptamers in the presence or in the absence of cytoskeleton pharmacological inhibitors. Our results showed that aptamers control cell morphology in a dose dependent manner (p < 0.0001). Simultaneously, when the inhibition of actin polymerization was induced, the control of cell morphology was attenuated (p < 0.0001), while no differences were detected when cells contractility was challenged (p > 0.05). Altogether, our data provide evidence that aptamers contribute to control fibronectin adsorption on biomaterials by preserving its conformation and thus function. Furthermore, our work provides a new insight into a new way to accurately tailor material surface bioactivity.

Tailoring the Interface of Biomaterials to Design Effective Scaffolds.

Tissue engineering (TE) is a multidisciplinary science, which including principles from material science, biology and medicine aims to develop biological substitutes to restore damaged tissues and organs. A major challenge in TE is the choice of suitable biomaterial to fabricate a scaffold that mimics native extracellular matrix guiding resident stem cells to regenerate the functional tissue. Ideally, the biomaterial should be tailored in order that the final scaffold would be (i) biodegradable to be gradually replaced by regenerating new tissue, (ii) mechanically similar to the tissue to regenerate, (iii) porous to allow cell growth as nutrient, oxygen and waste transport and (iv) bioactive to promote cell adhesion and differentiation. With this perspective, this review discusses the options and challenges facing biomaterial selection when a scaffold has to be designed. We highlight the possibilities in the final mold the materials should assume and the most effective techniques for its fabrication depending on the target tissue, including the alternatives to ameliorate its bioactivity. Furthermore, particular attention has been given to the influence that all these aspects have on resident cells considering the frontiers of materiobiology. In addition, a focus on chitosan as a versatile biomaterial for TE scaffold fabrication has been done, highlighting its latest advances in the literature on bone, skin, cartilage and cornea TE.