Evaluation of Human Gingival Fibroblasts (HGFs) Behavior on Innovative Laser Colored Titanium Surfaces.

The use of ytterbium laser to obtain colored titanium surfaces is a suitable strategy to improve the aesthetic soft tissue results and reduce implant failures in oral rehabilitation. To investigate the relationship between novel laser-colored surfaces and peri-implant soft tissues, Human Gingival Fibroblasts (HGFs) were cultured onto 12 colored titanium grade 1 light fuchsia, dark fuchsia, light gold, and dark gold disks and their viability (MTT Assay), cytotoxicity (lactate dehydrogenase release), and collagen I secretion were compared to the machined surface used as control. Optical and electronic microscopies showed a HGF growth directly correlated to the roughness and wettability of the colored surfaces. A higher viability percentage on dark fuchsia (125%) light gold (122%), and dark gold (119%) samples with respect to the machined surface (100%) was recorded. All specimens showed a statistically significant reduction of LDH release compared to the machined surface. Additionally, a higher collagen type I secretion, responsible for an improved adhesion process, in light fuchsia (3.95 µg/mL) and dark gold (3.61 µg/mL) compared to the machined surface (3.59 µg) was recorded. The in vitro results confirmed the innovative physical titanium improvements due to laser treatment and represent interesting perspectives of innovation in order to ameliorate aesthetic dental implant performance and to obtain more predictable osteo and perio-osteointegration long term implant prognosis.

Biologic Bone Behavior During the Osseointegration Process: Histologic, Histomorphometric, and SEM-EDX Evaluations.

This study evaluated bone behavior during dynamic osseointegration. A total of 12 implants were placed in sheep tibia and analyzed at 15, 30, 60, and 90 days. Quantitative and qualitative bone behaviors were evaluated with histologic, histomorphometric, Alizarin Red S, and SEM-EDX (scanning electron microscopy with energy-dispersive x-ray spectroscopy) analysis. Twenty microanalyses were performed in chambers 1, 3, and 5 (a chamber is the distinctive space/bone volume between two coils of the implant screw) in distinctive zones: the titanium-bone interface (zone A), the middle chamber-bone front (zone B), the bone-surgical threading interface (zone C), and native bone (zone D; used as a control). The dynamic osseointegration index (DOI) and bone quality index (BQI) with calcium/phosphorus (Ca/P) content were detected to evaluate the osseointegration quality, bone-to-implant contact (BIC), and bone density around implants. At 15 days, initial bone formation with osteoid matrix deposition and different color intensities were observed (means: BIC = 23.3% ± 3.9%; DOI = 1.55). SEM-EDX analysis showed low mineralized bone/bone marrow with a very low Ca/P mean value. At 30 days, high new bone deposition with higher color intensity in the crestal portion was recorded (BIC = 77.3% ± 0.4%; DOI = 2.58). At 90 days, tight BIC to the middle and apical implant portions were detected, as well as several osteon structures in the crestal portion (BIC = 86.4% ± 0.6%; DOI = 0.96). During all observed time periods, the BQI showed 25% more Ca/P in zone A. Greater maturation degree and lower BQI were seen at zone A compared to the other zones. After 15 and 30 days, zones B and C (except for P in zone B) showed BQIs slightly over 50% and around 75%, respectively, confirming a progressively higher degree of bone maturation that proceeds with the osseointegration process. After 90 days, the BQI values of zones B and C (greater than 70% in zone B and around 90% in zone C) confirmed the bone mineralization and maturation process and an acceleration of implant osseointegration, while a lower BQI value (25%) was recorded in zone A. This study shows osseointegration as a variable dynamic process with a higher bone deposition in contact with the implant surface during the early phase, while in the active and later osseointegration times, the bone quality maturation showed higher values only "at distance" (growth of native bone to the implant surface, observed later in the osseointegration process). After 3 months (before loading), the BQI evaluation was lower (25%) in zone A, confirming that the healing and maturation process of the bone cannot be considered complete.

Multidisciplinary investigation of an ancient renal stone in a mummy from Popoli, central Italy.

The renal stone found in the natural mummy of an anonymous nobleman dating to 19th century was investigated using advanced imaging modalities and analytic investigations. By this multidisciplinary approach we were able to identify the chemical components and their distribution throughout the sample. These results allowed to understand the lifestyle habits of the subject, as well as the exact pathogenesis of his disease.

Three-Dimensional (3D) Stereolithographic Tooth Replicas Accuracy Evaluation: In Vitro Pilot Study for Dental Auto-Transplant Surgical Procedures.

After immediate tooth extraction or after alveolar socket healing, tooth transplants are increasingly used for functional restoration of edentulous maxillary areas. Recent studies have shown the periodontal ligament (PDL) viability and the tooth housing time in the adapted neo-alveolus as key factors for transplantation success. During surgical time, 3D stereolithographic replicas are used for fitting test procedures. In this paper, the accuracy of 3D dental replicas, compared with the corresponding natural teeth, is assessed in surgical transplantation. Lamb skulls were selected and submitted to Cone Beam Computer Tomography (CBCT). Scanning information, converted into Standard Digital Imaging and Communications in Medicine (DICOM) and Standard Triangulation Language (STL), was sent to the Volux X-ray Centre for 3D replica printing. After the tooth extractions, all lambs' incisors were measured with a digital caliber and compared with the 3D replicas. Volume and dimensional error values were evaluated. All replicas showed macroscopically smaller volume (45.54%). Root replicas showed higher variations compared with the crown areas, with several unreplicated apical root areas. The cement-enamel junction tooth area was replicated quite faithfully, and the base area relative error showed 9.8% mean value. Even further studies with a larger number of replicas are needed. Data obtained confirmed high volumes of macroscopic discrepancies with several unreproduced apical root sites. The achieved accuracy (90.2%) confirmed that the 3D replicas cannot be used to reduce the surgical time during transplantation predictable procedures.

Effects of Novel Laser Dental Implant Microtopography on Human Osteoblast Proliferation and Bone Deposition.

PURPOSE: The aim of this study was to compare how two innovative laser titanium surfaces and sandblasted and acid-etched surfaces influence human osteoblast behavior during osteogenesis and the initial phases of bone deposition. MATERIALS AND METHODS: Human osteoblasts from human adipose stem cells were sorted by flow cytometric analysis and induced to differentiate. After 40 days, the osteogenic differentiation was detected by alizarin red staining, and the alkaline phosphatase (ALP) was evaluated with western blot (WB) and real-time reverse transcriptase-polymerase chain reaction (RT-PcR) analysis. After confluence, human osteoblasts were cultured onto two different innovative laser-obtained titanium surfaces (L1 and L2) and compared with one sandblasted and acid-etched (SBAE) surface as the control. At different times, human osteoblast behavior was evaluated with cell proliferation viability assay (MTT), scanning electron microscopy (SEM), energy-dispersive x-rays (EDAX), osteogenic markers with RT-PcR, and WB analysis of matrix extracellular phosphoglycoprotein (MEPE), ALP, and osteocalcin (OCN). RESULTS: After cell sorting, the human osteoblasts from human adipose stem cells showed increasing values of ALP mRNA and protein expression. The osteogenic differentiation was confirmed by quantitative alizarin red staining assay. Profilometric and SEM analysis showed relevant differences between SBAE, L1, and L2 specimens. After 20 days of culture onto titanium samples, SEM evaluation showed a small number of human osteoblasts and isolated sites of bone matrix deposition in SBAE specimens. At the same time, on L1 surfaces, only an osteoblast mono-layer with initial bone deposition was found, while on L2 specimens, there was a thick network with flattened large stellate cells, many cellular interconnections with strong titanium adhesion, and a large complex mineralized structure of crystal bone. After 20 days, for all titanium samples, human osteoblasts culturing EDAX analysis showed the absence of impurities and a higher bone matrix deposition in L2 specimens compared with L1 and SBAE samples. CONCLUSION: The innovative microtopography and nanotopography laser-induced surface showed high biocompatibility with primary human osteoblast cultures and the absence of impurities. The innovative laser texture was capable of influencing the osteogenic process, confirming the critical role of titanium surface characteristics in the cell adhesion and bone deposition during the early phases of osseointegration. The association of human adipose stem cells and titanium surfaces laser-induced with an innovative procedure could generate promising improvements and developments in orthopedics, maxillofacial, and dental implant surgery.

Esthetic and Physical Changes of Innovative Titanium Surface Properties Obtained with Laser Technology.

AIM: The purpose of the study was the evaluation of the esthetic and physical changes produced on colored titanium Grade 5 (Ti6Al4V) laser treated surfaces to be used in implant dentistry for esthetic success. MATERIALS AND METHODS: Colored titanium surfaces were obtained with laser treatment. The physical and topographic properties were evaluated by stereo, light, and electron microscopy and profilometric analyses. L*a*b* colorimetric coordinates were measured by spectrometry, and the superficial chemical characteristics were evaluated by energy dispersive X-ray analysis. RESULTS: Within the complete palette of titanium colors, pinks (P1-P2), incarnadine (I), and white (W) obtained by laser were selected. The topography, texture, hues, saturation, roughness, and porosity of the samples were compared with those of machined (M) and sand-blasted and etched (SBAE) control surfaces. P1, P2, and I, similar in hue and roughness (Ra @ 0.5 µm), had a microgroove spacing of 56 µm and a decreasing porosity. The W sample with a "checkerboard" texture and a light color (L* 96.31) was similar to the M samples (Ra = 0.32 µm), but different from SBAE (Ra = 1.41 µm, L* 65.47). DISCUSSION: The aspects of hard and soft tissue could result in an esthetic failure of the dental implant by showing the dark color of the fixture or abutment. The two different pinks and incarnadine surfaces showed favorable esthetic and physical features to promote dental implant success even in the maxillary anterior area with gingival recession, asymmetry, and deficiency. CONCLUSION: Titanium colored laser surfaces represent a valid alternative to those currently traditionally obtained and interesting and potential perspectives in the management of dental implants' esthetic failure.

Poly D,L-Lactide-Co-Glycolic Acid Grafting Material in Sinus Lift.

The poly D,L-lactide-co-glycolic acid (PLGA) is a copolymer used in many therapeutic devices for its high rates of biodegradability and biocompatibility. The principal aim of the research was to evaluate the new bone formation, after 16 (T1) and 28 weeks (T2), in sheep maxillary sinus lift in vivo model using PLGA.Computerized tomography analysis, X-ray microanalysis, and scanning electron microscope analysis of secondary electrons (SE) and the backscattered electrons (BSE) of the samples were detected.After 28 weeks, the computed tomography analysis showed a 22% increase of UH density in the grafting areas. The X-ray microanalysis of the samples showed calcium and phosphorus increase at T1 and T2 follow-up period and the carbon and oxygen concentration decrease. The SE evaluation showed a rapid superficial resorption of the biomaterials at T1 and a completely bone reorganization of biomaterial at T2. The BSE analysis confirmed the SE data and showed the direct and intimate contact between bone and PLGA with a higher calcification in T2 compared to T1.Certainly, still other experiments and a larger number of samples will be necessary to be analyzed to determine the behavior of the PLGA in the bone regeneration; however, the PLGA used in maxillary sinus lift animal model, seem to promote new bone formation that continues increase at 28 weeks after grafting.

Osteogenic Differentiation of Mesenchymal Stromal Cells: A Comparative Analysis Between Human Subcutaneous Adipose Tissue and Dental Pulp.

White adipose tissue is a source of mesenchymal stromal/stem cells (MSCs) that are actively studied for their possible therapeutic use in bone tissue repair/remodeling. To better appreciate the osteogenic potential of these cells, we compared some properties of MSCs from human subcutaneous adipose tissue [subcutaneous-adipose stromal cells (S-ASCs)] and dental pulp stem cell (DPSCs) of third-impacted molars, the latter representing a well-established MSC source. Both undifferentiated cell types showed similar fibroblast-like morphology and mesenchymal marker expression. However, undifferentiated S-ASCs displayed a faster doubling time coupled to greater proliferation and colony-forming ability than DPSCs. Also, the osteogenic differentiation of S-ASCs was greater than that of DPSCs, as evaluated by the higher levels of expression of early osteogenic markers Runt-related transcription factor-2 (RUNX2) and alkaline phosphatase at days 3-14 and of extracellular matrix mineralization at days 14-21. Moreover, S-ASCs showed a better colonization of the titanium scaffold. In addition, we investigated whether S-ASC osteogenic commitment was enhanced by adenosine A1 receptor (A1R) stimulation, as previously shown for DPSCs. Although A1R expression was constant during DPSC differentiation, it increased in S-ASC at day 21 from osteogenesis induction. Accordingly, A1R stimulation by the agonist 2-chloro-N6-cyclopentyl-adenosine, added to the cultures at each medium change, stimulated proliferation only in differentiating DPSC and enhanced the osteogenic differentiation earlier in DPSCs than in S-ASCs. These effects were counteracted by cell pretreatment with a selective A1R antagonist. Thus, our findings suggest that S-ASCs could be advantageously used in regenerative orthopedics/dentistry, and locally released or exogenously added purines may play a role in bone repair/remodeling, even though this aspect should be more thoroughly evaluated.

A comparison of bovine bone and hydroxyapatite scaffolds during initial bone regeneration: an in vitro evaluation.

OBJECTIVES: To evaluate the different behavior of 3-dimensional biomaterial scaffolds-Bovine Bone (BB; Bio-Oss) and Hydroxyapatite (HA; ENGIpore)-during initial bone healing and development. MATERIALS AND METHODS: Human dental papilla stem cells (hDPaSCs) were selected with FACsorter cytofluorimetric analysis, cultured with osteogenic medium, and analyzed with Alizarin red stained after differentiation. The obtained osteoblast-like cells (OCs) were cultured with BB and HA. alkaline phosphatase (ALP), OC, MEPE, and runt-related transcription factor 2 (RUNX2) expression markers were investigated performing Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analysis. After 40 days, samples were analyzed by light and electron microscopy. RESULTS: All the samples showed high in vitro biocompatibility and qualitative differences of OCs adhesion. RT-PCR and Western blot data exhibited similar marker rate, but ALP, OC, MEPE, and RUNX2expression, during initial healing and bone regeneration phase, was higher and faster in human dental papilla onto BB than in HA scaffolds. In biomaterials growth, RUNX2 seems to play an important role as a key regulator in human OCs from dental papilla bone development. CONCLUSION: Different surface BB scaffold characteristics seem to play a critical role in OCs differentiation showing different time of bone regeneration morphological characteristics as well as higher and faster levels of all observed markers.

Influence of novel nano-titanium implant surface on human osteoblast behavior and growth.

PURPOSE: The aim of the study is to investigate human osteoblast-like cell behavior and growth in the presence of 3 different titanium implant surfaces. MATERIALS: Human stem cells were first obtained and then sorted by fluorescence-activated cell sorter from mesenchymal stem cell clusters of human dental papilla. The obtained human dental papilla stem cells were induced to differentiate into osteoblast-like cells and were then analyzed by reverse transcriptase polymerase chain reaction and Western blot analyses. The cells proliferated and were cultured onto 3 different titanium discs (sandblasted, sandblasted and large-grit acid-etched, and full contact coverage [FCC]) and analyzed by scanning electron microscope. RESULTS: In all analyses samples, a high cell activity was observed, with typical osteoblast mature morphostructural response on rough surface. The high number of osteoblast-like cells was found on titanium FCC discs. At the same time, scanning electron microscope analysis confirmed the high biocompatibility of this surface. CONCLUSION: The rapid maturation of the osteoblast-like cells on FCC titanium surface suggests that this structure could play a central role during initial phases of bone healing processes.

Investigation by solid-phase microextraction and gas chromatography/mass spectrometry of secondary metabolites in lichens deposited on stone monuments.

Lichens are ubiquitous organisms formed by symbiotic associations of fungal hyphas and algae that also grow under often extreme environmental conditions. They produce secondary metabolites, the so-called lichen substances, whose structural characterization can give an important contribution to lichen taxonomy. Lichens are also widely employed as biomonitors of atmospheric pollution; being epiphyte organisms they tend, in fact, to accumulate exogenous compounds. Moreover, it could be questioned if the environmental stress alters their secondary metabolites production. Therefore, a new strategy for the analysis of the organic substances absorbed or metabolized by lichens has been developed. This method exploits the dry solid-phase microextraction (SPME) headspace technique coupled with gas chromatography/mass spectrometry (GC/MS). Lichens coating the stone surfaces of monuments, located in small towns between high mountains and far away from urban environments, have been investigated. In the field of cultural heritage, this study can contribute to the knowledge of the state of conservation of outdoor exposed historical monuments.