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STATEMENT OF PROBLEM: Intraoral repair techniques prevent unnecessary replacement of ceramic restorations, thereby increasing the survival rate. However, adhesion between ceramics and the composite resin is challenging and how different protocols influence adhesion is unclear. PURPOSE: The purpose of this systematic review and meta-analysis was to analyze the influence of different protocols on repairing glass-ceramic surfaces with composite resins. MATERIAL AND METHODS: PubMed, Scopus, ISI Web of Science, and Embase electronic databases were searched to select studies comparing bond strength values or survival rates of glass-ceramic repaired with composite resins using different surface treatment protocols. No publication year or language restriction was applied. Data sets were extracted from all included studies, and the mean differences calculated. A 95% confidence interval was calculated by using the random effect model (Rev Man 5.4). RESULTS: The search identified 5037 studies, and 165 were assessed for eligibility. Finally, 123 in vitro studies were included in the systematic review and 48 in the meta-analysis. Considering different glass-ceramics, bond strength tests, and aged or not aged specimens, 37 meta-analyses found the effect of repair protocols: only adhesive, silane plus adhesive alone or preceded by hydrofluoric (HF) acid, airborne-particle abrasion (APA) with Al2O3 particles, silica-coated APA (SCAPA), diamond rotary instrument (DRI), and laser irradiation (LI). CONCLUSIONS: For feldspathic porcelain, HF acid, APA, SCAPA, or DRI improved the repair micromechanical retention; applying silane is essential to HF-conditioned surfaces but the use of adhesive is optional when silane is applied. Results for leucite and lithium disilicate were inconclusive in terms of suggesting a treatment other than HF acid plus silane and adhesive applications.
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Pericytes and glial cells are known to collaborate in dental pulp tissue repair. Cell-based therapies that stimulate these stromal components may be of therapeutic relevance for partially vital dental pulp conditions. This study aimed to examine the early effect of photobiomodulation (PBM) in pericytes from experimentally injured pulp tissue. To accomplish this, we used the Nestin-GFP/NG2-DsRed mice, which could allow the identification of distinct pericyte phenotypes. We discovered the presence of two pericytes subsets within the dental pulp, the Nestin + NG2+ (type-2) and Nestin- NG2+ (type-1). Upon injury, PBM treatment led to a significant increase in Nestin+ cells and pericytes. This boost was mainly conferred by the more committed pericyte subset (NestinNG2+ ). PBM also stimulated terminal blood vessels sprouting adjacent to the injury site while maintaining signs of pulp vitality. In vitro, PBM induced VEGF upregulation, improved dental pulp cells proliferation and migration, and favored their mineralization potential. Herein, different subsets of perivascular cells were unveiled in the pulp tissue. PBM enhanced not only NG2+ cells but nestin-expressing progenitors in the injured dental pulp.
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Pulpa Dental/citología , Neuroglía , Pericitos , Animales , Ratones , Nestina/genética , TransgenesRESUMEN
Photobiomodulation therapy (PBMT) has been widely used to promote tissue repair. However, PBMT's critical roles in the epithelial and mesenchymal tissues interactions are still barely known. Herein, we investigated light parameters on challenged keratinocytes (KC)-i.e., cultivated under oxidative stress-solely or associated with fibroblasts (FB) in a co-culture system. Cells were treated with PBMT at the wavelength of 660 nm, at 20 mW and 0.71 W/cm2 . Three different energy densities were primarily evaluated on KC: 1 (1.4 s), 5 (7 s), and 50 J/cm2 (70 s). Next, KC and FB were co-cultured and assessed at 5 J/cm2 . This energy density was also tested in ex vivo murine skin samples. Our main data suggest that PBMT can increase cellular proliferation at low doses and cell migration in a biphasic mode (1 and 50 J/cm2 ), both further confirmed by the epidermal growth factor receptor ligand-amphiregulin-upregulation. IL-1RA mRNA-the IL-1ß (interleukin-1ß) receptor antagonist recognized to fasten wound repair-was upregulated in the co-culture system. Upon PBMT, the ex vivo findings showed a progressive increase in the epidermal thickness, although presenting qualitatively less differentiated epithelium than the control group. In conclusion, PBMT effects are dependent on the cellular interactions with the surrounding microenvironment. Ultimately, PBMT is anti-inflammatory and contributes to the expression of critical mediators of wound repair.
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Terapia por Luz de Baja Intensidad , Células Madre Mesenquimatosas , Animales , Fibroblastos/metabolismo , Queratinocitos , Ratones , Cicatrización de HeridasRESUMEN
Photobiomodulation therapy (PBMT) can improve processes relevant to tissue regeneration, such as survival, proliferation, migration, and differentiation of cells, including stem cells. Thus, PBMT could be applied as auxiliary therapy for tissue regeneration. Cell sheets (CSs) induced by vitamin C (VC) can generate large amount of cells, which would also be useful for tissue regeneration. VC and PBMT cause opposite effects on cell metabolism (e.g., VC is antioxidative, and PBMT generates reactive oxygen species); however, hDPSC CSs were formed when VC and PBMT+VC were applied. Thus, this study showed that PBMT does not interfere with the formation of cell sheets induced by VC. Additionally, PBMT improved the functional differentiation of the cells isolated from the CSs. Thus, due to the clinical benefits of PBMT, the association of this therapy with cell sheets seems promising for future applications in tissue regeneration.
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Ácido Ascórbico/farmacología , Diferenciación Celular/efectos de los fármacos , Pulpa Dental/efectos de los fármacos , Células Madre/efectos de los fármacos , Pulpa Dental/citología , Células Epiteliales/efectos de los fármacos , Humanos , Terapia por Luz de Baja Intensidad/métodos , Músculo Esquelético/efectos de los fármacos , Células Madre/citología , Cicatrización de Heridas/efectos de los fármacosRESUMEN
Photobiomodulation (PBM) therapy displays relevant properties for tissue healing and regeneration, which may be of interest for the tissue engineering field. Here, we show that PBM is able to improve cell survival and to interact with recombinant human Bone Morphogenetic Protein 4 (rhBMP4) to direct and accelerate odonto/osteogenic differentiation of dental derived mesenchymal stem cells (MSCs). MSCs were encapsulated in an injectable and thermo-responsive cell carrier (Pluronic® F-127) loaded with rhBMP4 and then photoactivated. PBM improved MSCs self-renewal and survival upon encapsulation in the Pluronic® F-127. In the presence of rhBMP4, cell odonto/osteogenic differentiation was premature and markedly improved in the photoactivated MSCs. An in vivo calvarial critical sized defect model demonstrated significant increase in bone formation after PBM treatment. Finally, a balance in the reactive oxygen species levels may be related to the favorable results of PBM and rhBMP4 association. PBM may act in synergism with rhBMP4 and is a promise candidate to direct and accelerate hard tissue bioengineering.
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Proteína Morfogenética Ósea 4/administración & dosificación , Portadores de Fármacos , Terapia por Luz de Baja Intensidad/métodos , Células Madre Mesenquimatosas/efectos de los fármacos , Células Madre Mesenquimatosas/efectos de la radiación , Poloxámero/química , Ingeniería de Tejidos/métodos , Andamios del Tejido , Adolescente , Adulto , Animales , Proteína Morfogenética Ósea 4/química , Regeneración Ósea , Diferenciación Celular/efectos de los fármacos , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de los fármacos , Proliferación Celular/efectos de la radiación , Autorrenovación de las Células/efectos de los fármacos , Autorrenovación de las Células/efectos de la radiación , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Humanos , Hidrogeles , Inyecciones , Láseres de Semiconductores , Terapia por Luz de Baja Intensidad/instrumentación , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/metabolismo , Ratones Desnudos , FN-kappa B/metabolismo , Osteogénesis/efectos de los fármacos , Osteogénesis/efectos de la radiación , Hueso Parietal/lesiones , Hueso Parietal/patología , Hueso Parietal/cirugía , Especies Reactivas de Oxígeno/metabolismo , Factores de Tiempo , Adulto JovenRESUMEN
Dermal wound healing is the process of repairing and remodeling skin following injury. Delayed or aberrant cutaneous healing poses a challenge for the health care system. The lack of detailed understanding of cellular and molecular mechanisms involved in this process hampers the development of effective targeted treatments. In a recent study, Parfejevs et al.-using state-of-the-art technologies, including in vivo sophisticated Cre/loxP techniques in combination with a mouse model of excisional cutaneous wounding-reveal that Schwann cells induce adult dermal wound healing. Strikingly, genetic ablation of Schwann cells delays wound contraction and closure, decreases myofibroblast formation, and impairs skin re-epithelization after injury. From a drug development perspective, Schwann cells are a new cellular candidate to be activated to accelerate skin healing. Here, we summarize and evaluate recent advances in the understanding of Schwann cells roles in the skin microenvironment.
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Células de Schwann/fisiología , Piel/lesiones , Cicatrización de Heridas/fisiología , Heridas y Lesiones/patología , Animales , Diferenciación Celular/fisiología , Células Cultivadas , Modelos Animales de Enfermedad , Ratones , Receptor Cross-Talk , Piel/patologíaRESUMEN
Cartilage tissue regeneration often presents a challenging clinical situation. Recently, it has been shown that Periodontal Ligament Stem Cells (PDLSCs) possess high chondrogenic differentiation capacity. In this study, we developed a stem cell delivery system based on alginate/hyaluronic acid (HA) loaded with TGF-ß1 ligand, encapsulating PDLSCs; and investigated the chondrogenic differentiation of encapsulated cells in alginate/HA hydrogel microspheres in vitro and in vivo. The results showed that PDLSCs, as well as human bone marrow mesenchymal stem cells (hBMMSCs), as the positive control, were stained positive for both toluidine blue and alcian blue staining, while exhibiting high levels of gene expression related to chondrogenesis (Col II, Aggrecan and Sox-9), as assessed via qPCR. The quantitative PCR analyses exhibited that the chondrogenic differentiation of encapsulated MSCs can be regulated by the modulus of elasticity of hydrogel delivery system, confirming the vital role of the microenvironment, and the presence of inductive signals for viability and differentiation of MSCs. In vivo, histological and immunofluorescence staining for chondrogenic specific protein markers confirmed ectopic cartilage-like tissue regeneration inside transplanted hydrogels. PDLSCs presented significantly greater capability for chondrogenic differentiation than hBMMSCs (P < 0.05). Altogether, our findings confirmed that alginate/HA hydrogels encapsulating PDLSCs are a promising candidate for cartilage regeneration.
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Alginatos/química , Diferenciación Celular/efectos de los fármacos , Condrocitos/efectos de los fármacos , Condrogénesis/efectos de los fármacos , Portadores de Fármacos/farmacología , Sistemas de Liberación de Medicamentos , Ácido Hialurónico/química , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacología , Ligamento Periodontal/efectos de los fármacos , Células Madre/efectos de los fármacos , Adolescente , Adulto , Alginatos/farmacología , Animales , Células Cultivadas , Condrocitos/fisiología , Condrogénesis/fisiología , Portadores de Fármacos/síntesis química , Portadores de Fármacos/química , Ácido Glucurónico/química , Ácido Glucurónico/farmacología , Ácidos Hexurónicos/química , Ácidos Hexurónicos/farmacología , Humanos , Ácido Hialurónico/farmacología , Hidrogel de Polietilenoglicol-Dimetacrilato/síntesis química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Ensayo de Materiales , Ratones , Ratones Desnudos , Microesferas , Ligamento Periodontal/citología , Ligamento Periodontal/fisiología , Células Madre/citología , Células Madre/fisiología , Adulto JovenRESUMEN
PURPOSE: Peri-implantitis is one of the most common inflammatory complications in dental implantology. Similar to periodontitis, in peri-implantitis, destructive inflammatory changes take place in the tissues surrounding a dental implant. Bacterial flora at the failing implant sites resemble the pathogens in periodontal disease and consist of Gram-negative anaerobic bacteria including Aggregatibacter actinomycetemcomitans (Aa). Here we demonstrate the effectiveness of a silver lactate (SL)-containing RGD-coupled alginate hydrogel scaffold as a promising stem cell delivery vehicle with antimicrobial properties. MATERIALS AND METHODS: Gingival mesenchymal stem cells (GMSCs) or human bone marrow mesenchymal stem cells (hBMMSCs) were encapsulated in SL-loaded alginate hydrogel microspheres. Stem cell viability, proliferation, and osteo-differentiation capacity were analyzed. RESULTS: Our results showed that SL exhibited antimicrobial properties against Aa in a dose-dependent manner, with 0.50 mg/ml showing the greatest antimicrobial properties while still maintaining cell viability. At this concentration, SL-containing alginate hydrogel was able to inhibit Aa growth on the surface of Ti discs and significantly reduce the bacterial load in Aa suspensions. Silver ions were effectively released from the SL-loaded alginate microspheres for up to 2 weeks. Osteogenic differentiation of GMSCs and hBMMSCs encapsulated in the SL-loaded alginate microspheres were confirmed by the intense mineral matrix deposition and high expression of osteogenesis-related genes. CONCLUSION: Taken together, our findings confirm that GMSCs encapsulated in RGD-modified alginate hydrogel containing SL show promise for bone tissue engineering with antimicrobial properties against Aa bacteria in vitro.
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Hidrogel de Polietilenoglicol-Dimetacrilato , Células Madre Mesenquimatosas , Periimplantitis/terapia , Alginatos , Antiinfecciosos , Humanos , OsteogénesisRESUMEN
Dental-derived mesenchymal stem cells (MSCs) provide an advantageous therapeutic option for tissue engineering due to their high accessibility and bioavailability. However, delivering MSCs to defect sites while maintaining a high MSC survival rate is still a critical challenge in MSC-mediated tissue regeneration. Here, we tested the osteogenic and adipogenic differentiation capacity of dental pulp stem cells (DPSCs) in a thermoreversible Pluronic F127 hydrogel scaffold encapsulation system in vitro. DPSCs were encapsulated in Pluronic (®) F-127 hydrogel and stem cell viability, proliferation and differentiation into adipogenic and osteogenic tissues were evaluated. The degradation profile and swelling kinetics of the hydrogel were also analyzed. Our results confirmed that Pluronic F-127 is a promising and non-toxic scaffold for encapsulation of DPSCs as well as control human bone marrow MSCs (hBMMSCs), yielding high stem cell viability and proliferation. Moreover, after 2 weeks of differentiation in vitro, DPSCs as well as hBMMSCs exhibited high levels of mRNA expression for osteogenic and adipogenic gene markers via PCR analysis. Our histochemical staining further confirmed the ability of Pluronic F-127 to direct the differentiation of these stem cells into osteogenic and adipogenic tissues. Furthermore, our results revealed that Pluronic F-127 has a dense tubular and reticular network morphology, which contributes to its high permeability and solubility, consistent with its high degradability in the tested conditions. Altogether, our findings demonstrate that Pluronic F-127 is a promising scaffold for encapsulation of DPSCs and can be considered for cell delivery purposes in tissue engineering.
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Hidrogeles , Células Madre Mesenquimatosas/citología , Poloxámero/química , Andamios del Tejido , Diente/citología , Adolescente , Adulto , Humanos , Masculino , Adulto JovenRESUMEN
Cisplatin plays a central role in cancer chemotherapy, but resistance to this drug remains a major obstacle in treatment. Drawbacks related to cisplatin failure may be associated with cell energy metabolism. This study investigated whether photobiomodulation (PBM) can potentiate the effects of cisplatin on keratinocytes (HaCat) and cancer cells (SCC25 and HN12). Cells were treated with laser (GaAlAs; 660 nm; 60 mW; 0.33 J; 2.14 W cm-2 ; 11.7 J cm-2 and 6 s) and cisplatin (7.8 µg mL-1 ) to evaluate cell viability, Ki-67, VEGF, TGF-ß1, EGF expression and ROS production. Observations were validated in the SCC25 cell lineage, where the type of cell death (necrosis/apoptosis) and the amount of ATP were assessed. Cell lineages showed increased sensitivity to cisplatin associated with PBM (Cis-PBM). Ki-67 was augmented in all cell lineages treated with Cis-PBM when compared to cisplatin alone (Cis). Cis or Cis-PBM significantly decreased VEGF expression in cancer cells, while no changes were seen in the expression of TGF-ß1 or EGF compared to control. ROS levels were similar in the Cis and Cis-PBM groups. Cells treated with Cis-PBM died by apoptosis, leading to greater consumption of ATP. These observations suggest that PBM may potentiate the effects of cisplatin, leading to increased drug cytotoxicity and enhanced cell death.
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Antineoplásicos/farmacología , Cisplatino/farmacología , Terapia por Luz de Baja Intensidad , Neoplasias de la Boca/patología , Línea Celular Tumoral , Terapia Combinada , HumanosRESUMEN
OBJECTIVES: To evaluate the antibacterial effects, cytotoxicity and microtensile bond strength of an adhesive containing low concentrations of silver nanoparticles (NAg). METHODS: Various concentrations of NAg (50, 100, 150, 200 and 250 ppm) were incorporated into the primer of the Scotchbond Multi-Purpose adhesive system (SBMP). Antibacterial activity was examined using a broth microdilution assay to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), agar diffusion assay and the MTT assay was used to examine the biofilm metabolic activity (S. mutans). The Microtensile Bond Test (µTBS) was performed after 24 h, followed by 6-months storage in distilled water. Cytotoxicity was assessed with an MTT reduction assay in human dental pulp stem cells viability after exposure to Nag-conditioned culture media during 0, 24, 48, and 72 h. The results were statistically analyzed (α ≤ 0.05). RESULTS: MIC was found between NAg 25 and 50 ppm MBC was determined at 50 ppm of NAg. Bacterial activity inhibition was higher than control in all NAg groups compared to control in agar diffusion assay. Biofilm inhibition was statistically higher in 250 ppm NAg than control. All NAg groups and SBMP presented similar cytotoxicity in each period. Adhesives with NAg 200 and 250 ppm and SBMP (control) presented the highest µTBS values, similar to that of SBMP control, in both instances (24 h and 6 months) (p > 0.05). CONCLUSIONS: The commercial primer containing NAg 250 ppm showed both antibacterial effect and reliable bond strength with no cytotoxicity increase. The addition of NAg to primers seems promising for the improvement of conventional dental adhesives efficacy. CLINICAL SIGNIFICANCE: The addition of low concentrations of NAg (250 ppm) to primers were effective to improve antibacterial effect preserving the bond strength and the biocompatibility of the commercial product. NAg/primer association could protect the tooth-adhesive interface increasing dental restoration longevity.
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Antibacterianos/toxicidad , Recubrimiento Dental Adhesivo , Cementos Dentales , Nanopartículas del Metal/toxicidad , Plata , Biopelículas , Humanos , Ensayo de Materiales , Cementos de Resina , Resistencia a la TracciónRESUMEN
Repair or regeneration of damaged nerves is still a challenging clinical task in reconstructive surgeries and regenerative medicine. Here, it is demonstrated that periodontal ligament stem cells (PDLSCs) and gingival mesenchymal stem cells (GMSCs) isolated from adult human periodontal and gingival tissues assume neuronal phenotype in vitro and in vivo via a subcutaneous transplantation model in nude mice. PDLSCs and GMSCs are encapsulated in a 3D scaffold based on alginate and hyaluronic acid hydrogels capable of sustained release of human nerve growth factor (NGF). The elasticity of the hydrogels affects the proliferation and differentiation of encapsulated MSCs within scaffolds. Moreover, it is observed that PDLSCs and GMSCs are stained positive for ßIII-tubulin, while exhibiting high levels of gene expression related to neurogenic differentiation (ßIII-tubulin and glial fibrillary acidic protein) via quantitative polymerase chain reaction (qPCR). Western blot analysis shows the importance of elasticity of the matrix and the presence of NGF in the neurogenic differentiation of encapsulated MSCs. In vivo, immunofluorescence staining for neurogenic specific protein markers confirms islands of dense positively stained structures inside transplanted hydrogels. As far as it is known, this study is the first demonstration of the application of PDLSCs and GMSCs as promising cell therapy candidates for nerve regeneration.