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(1) Background: Mucointegration seems to gain interest when talking about success in the maintenance of dental implants. As we well know, collagen fibres cannot be inserted due to the lack of root structure on the implant surface, so the structural integration of peri-implant tissues that provide a firm seal around implants seems to be of interest when it comes to ensuring the survival of dental implants. To achieve a good epithelial barrier, the physicochemical characteristics of the surfaces of the restorative materials are of vital importance; therefore, the objective of this study is to analyse the histological behaviour of the peri-implant soft tissues in three different restorative materials. (2) Methods: Histological analysis of biopsied peri-implant keratinised mucosa, inflammatory epithelium and connective tissue in contact with a reinforced composite (BRILLIANT Crios), a cross-linked polymethylmethacrylate (TELIO CAD), and a hybrid ceramic (Vita Enamic), restored on a customised Atlantis-type abutment (Dentsply Sirona) between 60 and 180 days after restoration. (3) Results: A greater number of cells per mm2 of keratinised epithelium is observed in the reinforced composite, which could indicate greater surface roughness with greater inflammatory response. In this way, the greater number of lymphocytes and the lateral cellular composition of the inflammatory cells confirm the greater inflammatory activity towards that material. The best material to rehabilitate was hybrid ceramic, as it shows a better cellular response. (4) Conclusions: Knowing the limitations of the proposed study, despite the fact that greater inflammation is observed in the reinforced composite relative to the other materials studied, no statistically significant differences were found.

Metallothioneins (MTs) are low molecular weight cysteine-rich proteins that can bind up to seven zinc ions. Among their numerous functions, MTs appear to act as protectors against oxidative and inflammatory injury. In our first published study, we reported downregulation of the isoforms MT1B (fold distance (FD) -2. 95; p = 0.0024), MT1F (FD -1.72; p = 0.0276), MT1X (FD -3.09; p = 0.0021), MT1H (FD -2.39; p = 0.0018), MT1M (FD -2.37; p = 0.0092), MT1L (FD -2. 55; p = 0.0048), MT1E (FD -2.71; p = 0.0014), MT2A (FD -2.35; p = 0.0072), MT1G (FD -2.24; p = 0.0118), and MT1A (FD -2.82; p = 0.0023) by comparing Down's syndrome patients with periodontal disease and implant failure to those without periodontal disease and with a positive progression of their implants. In this gene validation study, we intended to verify the results of our first gene expression analysis. Materials and Methods: In our retrospective case-control study, we performed retrotranscription (RT-qPCR) of 11 RNA-to-cDNA samples using the SuperScript™ VILO™ kit (50; reference 1,176,605) from Thermo Fisher. We conducted the study using the real-time PCR technique on the q-PCR ViiA 7 platform from Thermo Fisher. We chose the format of the Taqman Array Plate 16 Plus (reference 4,413,261) from Thermo Fisher, which accommodates 12 genes plus four controls (GAPDH, 18S, ACTB, and HPRT1). We conducted the analysis of the plates using the Thermo Fisher Cloud Web Software. Results: The results obtained through gene validation analysis show that in PD+RI+ patients, the genes encoding the isoforms MT1F (FD 0.3; p = 0.039), MT1X (FD 338; p = 0.0078), MT1E (FD 307; p = 0.0358), and MT2A (FD 252; p = 0.0428) continue to show downregulation, whereas MT1B (FD 2.75; p = 0.580), MT1H (FD 281; p = 0.152), MT1L (FD 354; p = 0.0965), and MT1G (FD 336; p = 0.0749) no longer show statistically significant results.

(1) Background: The use of physical barriers to prevent the invasion of gingival and connective tissue cells into bone cavities during the healing process is called guided bone regeneration. The objective of this in-vitro study was to compare the growth of human osteoblasts on Poly(Lactic–co–Glycolic) (PLGA) membranes modified with oxygen plasma and Hydroxyapatite (HA), silicon dioxide (SiO2), and titanium dioxide (TiO2) composite nanoparticles, respectively. (2) Methods: All the membranes received a common treatment with oxygen plasma and were subsequently treated with HA nanostructured coatings (n = 10), SiO2 (n = 10) and TiO2 (n = 10), respectively and a PLGA control membrane (n = 10). The assays were performed using the human osteoblast line MG-63 acquired from the Center for Scientific Instrumentation (CIC) from the University of Granada. The cell adhesion and the viability of the osteoblasts were analyzed by means of light-field microphotographs of each condition with the inverted microscope Axio Observer A1 (Carl Zeiss). For the determination of the mitochondrial energy balance, the MitoProbe™ JC-1 Assay Kit was employed. For the determination of cell growth and the morphology of adherent osteoblasts, two techniques were employed: staining with phalloidin-TRITC and staining with DAPI. (3) Results: The modified membranes that show osteoblasts with a morphology more similar to the control osteoblasts follow the order: PLGA/PO2/HA > PLGA/PO2/SiO2 > PLGA/PO2/TiO2 > PLGA (p < 0.05). When analysing the cell viability, a higher percentage of viable cells bound to the membranes was observed as follows: PLGA/PO2/SiO2 > PLGA/PO2/HA > PLGA/PO2/TiO2 > PLGA (p < 0.05), with a better energy balance of the cells adhered to the membranes PLGA/PO2/HA and PLGA/PO2/SiO2. (4) Conclusion: The membrane in which osteoblasts show characteristics more similar to the control osteoblasts is the PLGA/PO2/HA, followed by the PLGA/PO2/SiO2.

The main target of bone tissue engineering is to design biomaterials that support bone regeneration and vascularization. Nanostructured membranes of (MMA)1-co-(HEMA)1/(MA)3-co-(HEA)2 loaded with 5% wt of SiO2-nanoparticles (HOOC-Si-Membrane) were doped with zinc (Zn-HOOC-Si-Membrane) or doxycycline (Dox-HOOC-Si-Membrane). Critical bone defects were effectuated on six New Zealand-bred rabbit skulls and covered with the membranes. After six weeks, the bone architecture was evaluated with micro computed tomography. Three histological analyses were utilized to analyse bone regeneration, including von Kossa silver nitrate, toluidine blue and fluorescence. All membrane-treated defects exhibited higher number of osteocytes and bone perimeter than the control group without the membrane. Zn-HOOC-Si-Membranes induced higher new bone and osteoid area than those treated with HOOC-Si-Membranes, and control group, respectively. Zn-HOOC-Si-Membranes and Dox-HOOC-Si-Membranes attained the lowest ratio M1 macrophages/M2 macrophages. Dox-HOOC-Si-Membranes caused the lowest number of osteoclasts, and bone density. At the trabecular new bone, Zn-HOOC-Si-Membranes produced the highest angiogenesis, bone thickness, connectivity, junctions and branches. Zn-HOOC-Si-Membranes enhanced biological activity, attained a balanced remodeling, and achieved the greatest regenerative efficiency after osteogenesis and angiogenesis assessments. The bone-integrated Zn-HOOC-Si-Membranes can be considered as bioactive modulators provoking a M2 macrophages (pro-healing cells) increase, being a potential biomaterial for promoting bone repair.

Native collagen-based membranes are used to guide bone regeneration; but due to their rapid biodegradation, this treatment is often unpredictable. The purpose of this study was to investigate the biodegradability of natural collagen membranes. Three non-cross-linked resorbable collagen barrier membranes were tested: Derma Fina (porcine dermis), Evolution Standard (equine pericardium) and Duo-Teck (equine lyophilized collagen felt). 10 × 10 mm2 pieces of membranes were submitted to three different degradation procedures: (1) hydrolytic degradation in phosphate buffer solution, (2) enzyme resistance, using a 0.25% porcine trypsin solution, and (3) bacterial (Clostridium histolyticum) collagenase resistance test. Weight measurements were performed with an analytic microbalance. Thickness was measured with a digital caliper. Membranes were analyzed at different time-points, up to 21 d of immersion. A stereomicroscope was used to obtain membranes’ images. ANOVA and Student Newman Keuls were used for mean comparisons (p < 0.05), except when analyzing differences between time-points within the same membrane and solution where pair-wise comparisons were applied (p < 0.001). Derma Fina attained the highest resistance to all degradation challenges. Duo-Teck was the most susceptible membrane to degradation, complete degradation occurred as soon as 8 h. The bacterial collagenase solution performed as the most aggressive test as all membranes presented 100% degradation before 21 d.

Bruxism is an unconscious, involuntary and sustained motor activity that results in excessive teeth grinding or jaw clenching that could affect patients’ implants and rehabilitations. The aetiology for bruxism remains unknown, but it is known to involve multiple factors. The literature lacks studies on the possible effect of implant morphology on the resistance of the bone-implant osseointegrated interface when bruxism is present. Our objective is to assess the mechanical response of the bone-implant interface in bruxist patients whose implant prostheses are subjected to parafunctional cyclic loading over a simulated period of 10 years. A comparison was carried out between two implant types (M-12 and Astra Tech), and a pattern of bone loss was established considering both the stress state and the cortical bone surface loss as the evaluation criteria. Numerical simulation techniques based on the finite element analysis method were applied in a dynamic analysis of the received forces, together with a constitutive model of bone remodelling that alters the physical properties of the bone. The simulated cortical bone surface loss at the implant neck area was 8.6% greater in the Astra implant than in the M-12 implant. Compared to the M-12 implant, the higher sustained stress observed over time in the Astra implant, together with the greater cortical bone surface loss that occurred at its neck area, may be related to the major probability of failure of the prostheses placed over Astra implants in bruxist patients

The aim of our study is to evaluate different implant surface treatments using TiIV and TiV in in vitro and in vivo studies. An in vitro study was established comprising four study groups with treated and untreated TiIV titanium discs (TiIVT and TiIVNT) and treated and untreated TiV titanium discs (TiVT and TiVNT). The surface treatment consisted in a grit blasting treatment with alumina and double acid passivation to modify surface roughness. The surface chemical composition and the surface microstructure of the samples were analyzed. The titanium discs were subjected to cell cultures to determine cell adhesion and proliferation of osteoblasts on them. The in vivo study was carried out on the tibia of three New Zealand rabbits in which 18 implants divided into three experimental groups were placed (TiIVT, TiIVNT, and TiVT). Micro-computed tomography (micro-CT) was performed to determine bone density around the implants. The results showed that cell culture had minor adhesion and cell proliferation in TiIVT and TiVT within the first 6 and 24 h. However, no differences were found after 48 h. No statistically significant differences were found in the in vivo micro-CT and histological study; however, there was a positive trend in bone formation in the groups with a treated surface. Conclusions: All groups showed a similar response to in vitro cell proliferation cultures after 48 h. No statistically significant differences were found in the in vivo micro-CT and histological study.

Background: Sometimes dental implants seem to be the only therapeutic alternative for the oral rehabilitation of patients with Down syndrome, given that they usually lose all their teeth early due to suffering aggressive periodontitis and they do not usually have the skills required to wear removable prostheses. However, the evolution of dental implants in these patients shows very adverse results. It is possible that basal genetic alterations, or at least some characteristics of these, may underlie these clinical results. The metabolic pathway of metallothioneins, molecules with an important influence on bone metabolism, could be one of the said alterations. Aims: To determine whether the expression of metallothioneins (MTs) and their metabolic pathway may be identified and related to the periodontitis and lack of osseointegration of dental implants in Down syndrome patients. Materials and Methods: Retrospective study of cases and controls by comparing patients with Down syndrome, periodontal disease, and implant failure (four patients, test group) with patients with Down syndrome, without periodontal disease, and without implant failure after two years of following (seven patients, control group), by extracting peripheral blood at the time of the dental examination to extract RNA and its subsequent processing in relation to gene expression of the metabolic pathway of metallothioneins. Results: The results identified low expression in the group of patients with periodontal disease and implant failure of genes MT1E, MT1H, MT1X, MT1A, MT1B, MT1C, MT1L, MT2A, MT1M, and MT1G. Conclusions: The low MT1 and MT2 gene expression seems to be related to the onset of periodontal disease and implant rejection in Down syndrome patients, although more data are required to confirm whether this relationship is due to one of the two conditions, to both independently, or to the two jointly—this last option being indicated by our current study.

[Aim] Titanium implants are commonly used as replacement therapy for lost teeth and much current research is focusing on the improvement of the chemical and physical properties of their surfaces in order to improve the osseointegration process. TiO2, when it is deposited in the form of pillar array nanometric structures, has photocatalytic properties and wet surface control, which, together with UV irradiation, provide it with superhydrophilic surfaces, which may be of interest for improving cell adhesion on the peri-implant surface. In this article, we address the influence of this type of surface treatment on type IV and type V titanium discs on their surface energy and cell growth on them., [Materials and methods] Samples from titanium rods used for making dental implants were used. There were two types of samples: grade IV and grade V. In turn, within each grade, two types of samples were differentiated: untreated and treated with sand blasting and subjected to double acid etching. Synthesis of the film consisting of titanium oxide pillar array structures was carried out using plasma-enhanced chemical vapor deposition equipment. The plasma was generated in a quartz vessel by an external SLAN-1 microwave source with a frequency of 2.45 GHz. Five specimens from each group were used (40 discs in total). On the surfaces to be studied, the following determinations were carried out: (a) X-ray photoelectron spectroscopy, (b) scanning electron microscopy, (c) energy dispersive X-ray spectroscopy, (d) profilometry, (e) contact angle measurement or surface wettability, (f) progression of contact angle on applying ultraviolet irradiation, and (g) a biocompatibility test and cytotoxicity with cell cultures., [Results] The application of ultraviolet light decreased the hydrophobicity of all the surfaces studied, although it did so to a greater extent on the surfaces with the studied modification applied, this being more evident in samples manufactured in grade V titanium. In samples made in grade IV titanium, this difference was less evident, and even in the sample manufactured with grade IV and SLA treatment, the application of the nanometric modification of the surface made the surface optically less active. Regarding cell growth, all the surfaces studied, grouped in relation to the presence or not of the nanometric treatment, showed similar growth., [Conclusions] Treatment of titanium oxide surfaces with ultraviolet irradiation made them change temporarily into superhydrophilic ones, which confirms that their biocompatibility could be improved in this way, or at least be maintained.

The novelty of this study is the addition of an ultrathin layer of nanostructured hydroxyapatite (HA) on oxygen plasma modified poly(lactic–co–glycolic) (PLGA) membranes (PO2) in order to evaluate the efficiency of this novel material in bone regeneration. Methods: Two groups of regenerative membranes were prepared: PLGA (control) and PLGA/PO2/HA (experimental). These membranes were subjected to cell cultures and then used to cover bone defects prepared on the skulls of eight experimental rabbits. Results: Cell morphology and adhesion of the osteoblasts to the membranes showed that the osteoblasts bound to PLGA were smaller and with a lower number of adhered cells than the osteoblasts bound to the PLGA/PO2/HA membrane (p < 0.05). The PLGA/PO2/HA membrane had a higher percentage of viable cells bound than the control membrane (p < 0.05). Both micro-CT and histological evaluation confirmed that PLGA/PO2/HA membranes enhance bone regeneration. A statistically significant difference in the percentage of osteoid area in relation to the total area between both groups was found. Conclusions: The incorporation of nanometric layers of nanostructured HA into PLGA membranes modified with PO2 might be considered for the regeneration of bone defects. PLGA/PO2/HA membranes promote higher osteosynthetic activity, new bone formation, and mineralisation than the PLGA control group.