RESUMEN

Cellular therapy using adipose tissue-derived mesenchymal stromal cells (at-MSCs) has garnered attention for the treatment of bone defects. Therefore, preconditioning strategies to enhance the osteogenic potential of at-MSCs could optimize cell therapy outcomes, and photobiomodulation (PBM) therapy has emerged as an effective, noninvasive, and low-cost alternative. This study explored the impacts of PBM on at-MSCs differentiation and the subsequent repair of bone defects treated with cell injection. Rat at-MSCs were cultured and irradiated (at-MSCsPBM) following the PBM protocol (660 nm; 20 mW; 0.714 W/cm2; 0.14 J; 5 J/cm2). Cellular differentiation was assessed based on the expression of gene and protein markers. Reactive oxygen species (ROS) were detected using fluorescence. At-MSCsPBM were injected into 5-mm calvarial lesions, and bone formation was analyzed using micro-CT and histological evaluations. At-MSCs were used as control. Data were analyzed using the ANOVA or t-test. At-MSCsPBM exhibited high levels of gene and protein runt-related transcription factor-2 (Runx2) and alkaline phosphatase (Alp) expression. PBM increased ALP activity and significantly reduced ROS levels. In addition, PBM increased the expression of Wnt pathway-associated genes. In vivo, there was an increase in the morphometric parameters, including bone volume, percentage of bone volume, bone surface area, and trabecular number, in at-MSCsPBM-treated defects compared with those in the control. These findings suggest that PBM enhances the osteogenic potential of at-MSCs, thereby supporting the advancement of improved cellular therapies for bone regeneration.

RESUMEN

Osteomyelitis is an inflammation of bone tissue usually caused by pyogenic bacteria. The most recurrent clinical approach consists of bone debridement followed by parenteral administration of antibiotics. However, systemic antibiotic treatment has limitations regarding absorption rate and bioavailability over time. The main challenge of osteomyelitis treatment consists of coupling the persistent infection treatment with the regeneration of the bone debrided. In this work, we developed an injectable drug delivery system based on poloxamer 407 hydrogel containing undoped Mg, Zn-doped tricalcium phosphate (ß-TCP), and teicoplanin, a broad-spectrum antibiotic. We evaluated how the addition of teicoplanin and ß-TCP affected the micellization, gelation, particle size, and surface charge of the hydrogel. Later, we studied the hydrogel degradation and drug delivery kinetics. Finally, the bactericidal, biocompatibility, and osteogenic properties were evaluated through in vitro studies and confirmed by in vivo Wistar rat models. Teicoplanin was found to be encapsulated in the corona portions of the hydrogel micelles, yielding a bigger hydrodynamics radius. The encapsulated teicoplanin showed a sustained release over the evaluated period, enough to trigger antibacterial properties against Gram-positive bacteria. Besides, the formulations were biocompatible and showed bone healing ability and osteogenic properties. Finally, in vivo studies confirmed that the proposed locally injected formulations yielded osteomyelitis treatment with superior outcomes than parenteral administration while promoting bone regeneration. In conclusion, the presented formulations are promising drug delivery systems for osteomyelitis treatment and deserve further technological improvements.

Asunto(s)

Antibacterianos , Fosfatos de Calcio , Hidrogeles , Osteogénesis , Osteomielitis , Ratas Wistar , Teicoplanina , Osteomielitis/tratamiento farmacológico , Osteomielitis/microbiología , Animales , Fosfatos de Calcio/química , Teicoplanina/administración & dosificación , Teicoplanina/farmacología , Teicoplanina/química , Antibacterianos/administración & dosificación , Antibacterianos/farmacología , Antibacterianos/química , Ratas , Hidrogeles/química , Hidrogeles/administración & dosificación , Osteogénesis/efectos de los fármacos , Sistemas de Liberación de Medicamentos/métodos , Humanos , Staphylococcus aureus/efectos de los fármacos , Poloxámero/química

RESUMEN

Background: Osteoradionecrosis (ORN) of the jaws is a late complication after radiotherapy to head and neck cancer. Objective: To describe a rare case of ORN of the torus mandibularis that was successfully managed exclusively with antimicrobial photodynamic therapy (aPDT). Case report: A 72-year-old man presented an exposed necrotic bone observed in the torus mandibularis, extending to the lingual alveolar ridge with no edema nor suppuration. The treatment provided a noninvasive treatment leading to spontaneous sequestrectomy of the torus in 2 weeks with complete mucosal repair in 5 weeks and absence of lesion signs and/or symptoms even after 6 months of follow-up. Conclusions: The aPDT indicated to be a satisfactory treatment for ORN affecting torus mandibularis, a region with surgical limitations, avoiding surgery.

Asunto(s)

Osteorradionecrosis , Fotoquimioterapia , Humanos , Masculino , Anciano , Osteorradionecrosis/etiología , Osteorradionecrosis/terapia , Osteorradionecrosis/tratamiento farmacológico , Fármacos Fotosensibilizantes/uso terapéutico , Enfermedades Mandibulares/etiología , Enfermedades Mandibulares/terapia , Enfermedades Mandibulares/tratamiento farmacológico

RESUMEN

AIM: This study aimed to evaluate the cytotoxicity, biocompatibility and osteoinductive profile of a mineral trioxide aggregate (MTA)-hydrogel-based material (MTA Flow) in comparison with MTA Angelus. METHODOLOGY: Cell viability was evaluated in human periodontal ligament stem cells (hPDLSCs) using the methyl-thiazol-tetrazolium (MTT) colourimetric assay. Polyethylene tubes containing the tested materials and empty polyethylene tubes (control) were implanted in the subcutaneous tissue of Wistar rats. Cellular (lymphocyte infiltration) and extracellular events (ECM; collagen fibres) were analysed in histological sections. Immunohistochemical (collagen I, osteopontin, bone sialoprotein, bone morphogenetic protein4) analyses were also performed. RESULTS: At 24, 48 and 72 h, all tested groups showed cell viability similar to control (p > .05). Regarding biocompatibility, all groups showed similar cellular events represented by a slight inflammatory reaction characterized by hyperaemia and a mild lymphocytic inflammatory infiltrate. The analysis of lymphocytes during the time showed a decrease in these cells in the control group and a significant interaction between MTA Angelus and control (p < .001), with MTA Angelus showing a more extensive inflammatory infiltrate. Regarding fibres, an increase in content was observed in all groups during the experimental time (7, 30 and 60 days), however, no difference was detected among the experimental groups (p = .063). After 60 days, the immunoexpression of bone matrix proteins in the MTA Flow group was similar to or higher than that observed in the MTA Angelus and in the control group. CONCLUSIONS: MTA Flow showed a non-cytotoxic behaviour, biocompatibility and ability to stimulate tissue mineralization.

Asunto(s)

Materiales Biocompatibles , Materiales de Obturación del Conducto Radicular , Ratas , Animales , Humanos , Ratas Wistar , Materiales Biocompatibles/farmacología , Compuestos de Calcio/farmacología , Hidrogeles , Óxidos/farmacología , Silicatos/toxicidad , Cementos Dentales , Cementos de Ionómero Vítreo , Colágeno , Polietilenos , Combinación de Medicamentos , Compuestos de Aluminio/toxicidad , Materiales de Obturación del Conducto Radicular/toxicidad , Ensayo de Materiales

RESUMEN

This study aimed to evaluate the antibacterial effect, cytotoxicity, and microtensile bond strength of an adhesive system containing silver nanoparticles (NAg). NAg was synthesized and incorporated (500 and 1000 ppm) into Scotchbond Multi-Purpose (SBMP) primer and bond. A microtensile bond test (µTBS) was performed after 24 h and 1 year. The adhesive interface was characterized using a confocal Raman microscope. The antibacterial activity was assessed using agar diffusion and biofilm inhibition assays (S. mutans). MTT assay was used to assess the cytotoxicity of NAg-conditioned culture media on human dental pulp stem cells (hDPSCs). The results were statistically analyzed using analysis of variance and Tukey's tests (α = .01). Incorporating 500 and 1000 ppm of NAg in the SBMP did not affect the µTBS after 24 h (p > 0.05). However, in the 1 year evaluation, 500 ppm presented the highest µTBS values (p < 0.05). The addition of NAg at 500 and 1000 ppm in the primer and bond led to larger inhibition halos and colony-forming units than the control (p < 0.05). For the unpolymerized and polymerized groups, the combination of primer and bond presented the highest cytotoxic effects on hDPSCs (p < 0.05). In conclusion, incorporating 500 or 1000 ppm of NAg into an etch-and-rinse adhesive system led to an antibacterial effect without altering the cytotoxicity. SBMP at 500 ppm presented a higher µTBS at 1 year.

Asunto(s)

Recubrimiento Dental Adhesivo , Nanopartículas del Metal , Humanos , Nanopartículas del Metal/química , Plata/farmacología , Plata/química , Cementos de Resina/farmacología , Cementos de Resina/química , Antibacterianos/farmacología , Resistencia a la Tracción , Cementos Dentales/farmacología , Cementos Dentales/química , Ensayo de Materiales , Recubrimientos Dentinarios/farmacología , Recubrimientos Dentinarios/química , Dentina

RESUMEN

This systematic review aimed to answer the research focused question: What are the effects of photobiomodulation (PBM) therapy on bone healing after ionizing irradiation in animal models? The EMBASE, LILACS, LIVIVO, PubMed, Scopus, and Web of Science databases, including gray literature, were searched using the following keywords: "Head and Neck Neoplasms"; "Ionizing Radiation"; "Low-Level Light Therapy"; and "Bone regeneration", focusing on the primary studies that assessed the effects of PBM therapy on animal models of irradiated bone. Six studies have met the eligibility criteria and presented an overall regular quality according to the risk of bias assessment tools. All the studies utilized rat animal model and near-infrared laser PBM at low power output setting. Most of the studies showed increased new bone formation, osteocytes, osteoblasts, and vascularization networking, as a result of PBM therapy. However, only one out of the six studies has not shown any differences in bone healing in both lased and non-lased animal groups. Nevertheless, PBM therapy is a potential tool to improve bone healing induced by ionizing radiation. However, due to the scarce number of studies and the great variability of laser parameters and treatment protocols, a clear conclusion cannot be drawn. Hence, extensive preclinical in vivo studies are warranted to ensure these beneficial effects have been addressed prior to translational clinical trials.

Asunto(s)

Neoplasias de Cabeza y Cuello , Terapia por Luz de Baja Intensidad , Ratas , Animales , Terapia por Luz de Baja Intensidad/métodos , Regeneración Ósea , Cicatrización de Heridas , Rayos Láser

RESUMEN

OBJECTIVES: To assess the effect of an experimental 58S bioactive glass on dentin permeability (dP) and erosive tooth wear (dentin surface loss - dSL). METHODS: 58S bioactive glass was synthetized using a sol-gel methodology, following by lyophilization and calcination, then mixed with phosphoric acid to obtain a paste (BGP). Forty-eight dentin disks (1 mm-thick) were used for dP, and 48 dentin slabs (3 mm × 3 mm) for dSL, which were assessed at three time intervals: post-EDTA (5 min in 17% EDTA solution); post-treatment (C: distilled water; BGP: experimental bioactive glass paste; NP: Nupro prophylaxis paste; CXT: Clinpro XT varnish); and post-erosive/abrasive cycling. Data were statistically analyzed (α=0.05). RESULTS: For dP and dSL, Groups did not differ significantly post-EDTA (p>0.05). Post-treatment, all groups showed lower dP than C (p<0.05), without differing significantly among them. For the dSL analysis, Groups C, BGP and NP did not differ significantly, showing lower values than CXT (p<0.05). Post-cycling, C continued to show the highest dP (p<0.05). Specimens from Group CXT had the lowest dP and did not differ from NP (p=0.86) which did not differ from BGP (p=0.193). For C and BGP, dP value was higher post-cycling than post-treatment (p<0.05). For NP and CXT, these experimental times did not differ (p>0.05). Post-cycling, dSL for C, BGP and NP did not differ significantly; values were higher than those for CXT (p<0.05). CONCLUSIONS: BGP reduced dP after application, with a reduced effect after cycling. Nonetheless, it was not able to protect dentin against erosive tooth wear. CLINICAL SIGNIFICANCE: Minimizing dentin hypersensitivity is a challenge in the field of dentistry. The development of alternative products with potential to obliterate dentinal tubules and provide resistance to chemical/mechanical stimuli is, thus, highly desirable. We have proposed a material able to reduce dentin permeability, which has emerged as a promising alternative for this purpose.

Asunto(s)

Sensibilidad de la Dentina , Atrición Dental , Erosión de los Dientes , Desgaste de los Dientes , Humanos , Sensibilidad de la Dentina/tratamiento farmacológico , Ácido Edético/farmacología , Ácido Edético/uso terapéutico , Dentina , Microscopía Electrónica de Rastreo , Erosión de los Dientes/prevención & control

RESUMEN

Melanoma is a highly aggressive skin cancer that requires new approaches for its management. Low-level laser therapy, currently named photobiomodulation therapy (PBM), has been used to improve different conditions but its effects and safe use on melanoma remain unexplored. Herein, we investigated the PBM impact on melanoma cells differing by pigmentation using near-infrared (NIR) and red lasers in vitro. In vivo, we evaluated the effects of the red laser on melanoma-bearing mice. Amelanotic (SK-MEL-37) and melanotic (B16F10) cells were exposed in vitro to a NIR (780 nm, 40 mW) or a red laser (660 nm, 40 mW) in 3 different light doses: 30, 90, and 150 J/cm2 and responses were assessed regarding mitochondrial activity, invasiveness, migration, and VEGF production. In vivo, melanoma-bearing mice received the red laser delivering 150 J/cm2 directly to the tumor on 3 consecutive days. Mice were monitored for 15 days regarding tumor progression and mouse survival. We noticed that amelanotic cells were unresponsive to NIR light. In contrast, NIR irradiation at 30 J/cm2 promoted an increase in the invasiveness of pigmented cells, even though all light doses have inhibited cell migration. Regarding the red laser on pigmented cells, the highest light dose (150 J/cm2) decreased the VEGF production and migration. In vivo, melanoma-bearing mice treated with red laser showed smaller tumor volume and longer survival than controls. We conclude that PBM appears to be safe for amelanotic non-pigmented melanoma but triggers different responses in melanotic pigmented cells depending on light parameters. Additionally, a high dose of red laser impairs the invasive behavior of melanoma cells, probably due to the decrease in VEGF synthesis, which may have contributed to tumor arrest and increased mouse survival. These findings suggest that red laser therapy could be a new ally in the supportive care of melanoma patients.

Asunto(s)

Terapia por Luz de Baja Intensidad , Melanoma , Animales , Luz , Melanoma/radioterapia , Ratones , Pigmentación , Factor A de Crecimiento Endotelial Vascular

RESUMEN

OBJECTIVE: This study aimed to assess the genotoxicity and cytotoxicity of Sealer Plus BC (SBC), AH Plus (AHP) and MTA Fillapex (MTF). METHODS: Human periodontal ligament dental stem cells (hPDLSCs) from third molars were isolated and cultured in a clonogenic medium. Cells were maintained in an incubator, and cell growth was monitored daily. hPDLSCs were characterised under flow cytometry and stem cell surface markers. The tested groups were a control group, SBC, AHP and MTF. Each sealer was prepared according to the manufacturer's instructions and placed in a clonogenic medium to produce a conditioned media. Conditioned media were then diluted to 10% to be placed in contact with culture cells in cell viability assay afterwards. The cells were harvested and plated into 96 wells culture plates. Genotoxicity was assessed by evaluation of micronucleus formation and cytotoxicity by MTT-based assay. All experiments were performed in triplicate. Data normality was verified by the Kolmogorov-Smirnov test. Statistical analysis for genotoxicity was performed with Kruskal-Wallis and Dunn's tests and two-way ANOVA for cytotoxicity, both with a significance level of 5%. RESULTS: Cells expressed typical levels of mesenchymal stem cell surface markers. No differences in the number of micronuclei were observed among all groups (P>0.05). In all periods analysed (24, 48, and 72 h), the sealers presented statistically different results for cell viability (P<0.05), with SBC presenting the lowest cytotoxicity, followed by the control group, MTF, and AHP. CONCLUSION: All sealers presented low genotoxicity, and Sealer Plus BC presented the lowest cytotoxicity.

Asunto(s)

Ligamento Periodontal , Materiales de Obturación del Conducto Radicular , Compuestos de Calcio , Medios de Cultivo Condicionados , Humanos , Materiales de Obturación del Conducto Radicular/toxicidad , Silicatos , Células Madre

RESUMEN

INTRODUCTION: The light-emitting diode (Led) in the violet spectrum associated or not with hydrogen peroxide (HP) has been suggested as a promising technique for dental bleaching. Violet led has a wavelength of 405-410 nm, which is very close to that of ultraviolet (UV) radiation, and this has raised biological safety concerns. AIM: To investigate the effectiveness of the violet led dental bleaching technique by evaluating color parameters, enamel surface microhardness, and biological safety analysis. METHODS: One hundred bovine dental blocks were divided into groups according to the bleaching technique (G1 - only HP; G2 - HP associated with blue led; G3 - only blue led; G4 - HP associated with a violet led; and G5 - only violet led). The color analysis (ΔE, ΔL, and WID) and enamel surface microhardness were assessed before and after bleaching (immediately, 5, 14, and 30 days). The biological safety of the violet led irradiation was assessed by measuring the number of micronuclei formed in human cells in culture in response to irradiation. Data analysis included Kruskal-Wallis test, Friedman test, and Mann-Whitney test. RESULTS: In groups G4 and G5 there was the formation of precipitates on the enamel surface. At the time of 14 days, it was observed that the G2 group had lower values of microhardness than G5. ΔL and ΔE showed differences between groups in experimental times. Mean percentages of micronuclei occurrence were similar in the control group and the violet led group. CONCLUSION: The violet led irradiation can be applied for dental bleaching because this approach produces significant color changes preserving tooth enamel integrity and causes no genotoxic effects on vital cells.

Asunto(s)

Fotoquimioterapia , Blanqueadores Dentales , Blanqueamiento de Dientes , Animales , Bovinos , Humanos , Peróxido de Hidrógeno , Fotoquimioterapia/métodos , Blanqueamiento de Dientes/métodos , Rayos Ultravioleta

RESUMEN

Diabetes mellitus (DM) is a chronic metabolic disease that affects bone metabolism, which can be related to a reduced osteogenic potential of bone marrow mesenchymal stem cells (BM-MSCs). MSCs from diabetic rats (dBM-MSC) have shown a tendency to differentiate towards adipocytes (AD) instead of osteoblasts (OB). Since photobiomodulation (PBM) therapy is a non-invasive treatment capable of recovering the osteogenic potential of dBM-MSCs, we aimed to evaluate whether PBM can modulate MSC's differentiation under hyperglycemic conditions. BM-MSCs of healthy and diabetic rats were isolated and differentiated into osteoblasts (OB and dOB) and adipocytes (AD and dAD). dOB and dAD were treated with PBM every 3 days (660 nm; 5 J/cm2; 0.14 J; 20 mW; 0.714 W/cm2) for 17 days. Cell morphology and viability were evaluated, and cell differentiation was confirmed by gene expression (RT-PCR) of bone (Runx2, Alp, and Opn) and adipocyte markers (Pparγ, C/Ebpα, and C/Ebpß), production of extracellular mineralized matrix (Alizarin Red), and lipid accumulation (Oil Red). Despite no differences on cell morphology, the effect of DM on cells was confirmed by a decreased gene expression of bone markers and matrix production of dOB, and an increased expression of adipocyte and lipid accumulation of dAD, compared to heatlhy cells. On the other hand, PBM reversed the effects of dOB and dAD. The negative effect of DM on cells was confirmed, and PBM improved OB differentiation while decreasing AD differentiation, driving the fate of dBM-MSCs. These results may contribute to optimizing bone regeneration in diabetic patients.

Asunto(s)

Diabetes Mellitus Experimental , Hiperglucemia , Células Madre Mesenquimatosas , Adipocitos , Animales , Células de la Médula Ósea , Diferenciación Celular , Células Cultivadas , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Experimental/radioterapia , Hiperglucemia/metabolismo , Hiperglucemia/radioterapia , Lípidos , Osteoblastos , Osteogénesis/genética , Ratas

RESUMEN

This systematic review aimed to assess whether the use of autologous platelet concentrates immediately after tooth extraction would prevent ORN in patients treated with radiotherapy (RT) for head and neck cancer (HNC). MEDLINE, Embase, Ovid, Scopus, Web of Science, CENTRAL Cochrane, and OpenGrey databases were searched (up to, and including, June 2021) by two independent reviewers to identify studies, followed by further manual search. As inclusion and exclusion criteria for the studies, only controlled clinical trials (randomized or not) were considered. The risk of bias of each included study was assessed using the Cochrane Risk of Bias Tool. Of the 129 potentially eligible studies, only 2 were included, both randomized controlled clinical trials; however, one used platelet-rich plasma and another leukocyte- and platelet-rich fibrin. Overall, both autologous platelet concentrates had no effect on the outcome assessed. The use of autologous platelet concentrates seems not to be beneficial for ORN prevention following tooth extractions in HNC patients treated with RT; however, according to the available evidence, a reliable statement cannot be made.

Asunto(s)

Osteorradionecrosis , Fibrina Rica en Plaquetas , Plasma Rico en Plaquetas , Humanos , Osteorradionecrosis/prevención & control , Extracción Dental

RESUMEN

OBJECTIVE: This study aimed to conduct a systematic review of the use of a cell sheet formed by mesenchymal stem cells derived from dental tissues (ddMSCs) for periodontal tissue regeneration in animal models in comparison with any other type of regenerative treatment. DESIGN: PubMed and Scopus databases were searched for relevant studies up to December 2020. The review was conducted based on the Preferred Reporting Items for Systematic Reviews and Meta-analysis guidelines. RESULTS: Of the 1542 potentially relevant articles initially identified, 33 fulfilled the eligibility criteria and were considered for this review. Even with a wide variety of selected study methods, the periodontal tissue was always regenerated; this indicates the potential for the use of these cell sheets in the future of periodontics. However, this regeneration process is not always complete. CONCLUSION: Despite the implantation, ddMSCs sheets have a great potential to be used in the regeneration of periodontal tissue. More in vivo studies should be conducted using standardized techniques for cell sheet implantation to obtain more robust evidence of the relevance of using this modality of cell therapy for periodontal tissue regeneration.

Asunto(s)

Células Madre Mesenquimatosas , Ligamento Periodontal , Animales , Biotecnología , Periodoncio , Ingeniería de Tejidos , Cicatrización de Heridas

RESUMEN

BACKGROUND: Methylmercury (MeHg) is a potent toxicant able to harm human health, and its main route of contamination is associated with the consumption of contaminated fish and other seafood. Moreover, dental amalgams are also associated with mercury release on human saliva and may contribute to the accumulation of systemic mercury. In this way, the oral cavity seems to be the primary location of exposure during MeHg contaminated food ingestion and dental procedures but there is a lack of literature about its effects on dental tissues and the impact of this toxicity on human health. In this way, this study aimed to analyze the effects of different doses of MeHg on human dental pulp stem cells after short-term exposure. METHODS: Dental pulp stem cells from human exfoliated deciduous teeth (SHED) were treated with 0.1, 2.5 and 5 µM of MeHg during 24 h. The MeHg effects were assessed by evaluating cell viability with Trypan blue exclusion assay. The metabolic viability was indirectly assessed by MTT reduction assay. In order to evaluate an indicative of antioxidant defense impairment, cells exposed to 0.1 and 5 µM MeHg were tested by measuring glutathione (GSH) level. RESULTS: It was observed that cell viability decreased significantly after exposure to 2.5 and 5 µM of MeHg, but the metabolic viability only decreased significantly at 5 µM MeHg exposure, accompanied by a significant decrease in GSH levels. These results suggest that an acute exposure of MeHg in concentrations higher than 2.5 µM has cytotoxic effects and reduction of antioxidant capacity on dental pulp stem cells.

RESUMEN

BACKGROUND: Dental trauma, restorative operative procedures and/or caries lesions can expose the dental pulp. Facing this clinical condition, where the maintenance of the dentin-pulp complex vitality is imperative, is challenging in Dentistry. Dental pulp stem cells conditioned medium contains trophic factors that could help in this task. This in vivo pilot study aimed to evaluate the effects of the human dental pulp stem cells conditioned medium on the dental pulp tissue response to vital pulp therapy. MATERIAL AND METHODS: Concentrated conditioned medium was obtained by incubating characterized human dental pulp stem cells with fresh culture medium. Pulp exposures performed at the first upper molars (n = 20) of Wistar rats were directly capped with: MTA or MTA + Conditioned Medium. Four and 8 weeks later, the samples were qualitatively analyzed in histological sections (H&E). RESULTS: When the conditioned medium was associated with MTA, there were a high percentage of samples presenting formation of dentin bridges and small percentage of pulp tissue with inflammatory signs in both experimental times. The conditioned medium improved the organization of the newly formed hard tissue. CONCLUSIONS: The association of dental pulp stem cell conditioned medium with MTA showed beneficial effects on dentin-pulp complex regeneration and has promising potential for studies in regenerative dentistry.

Asunto(s)

Medios de Cultivo Condicionados/farmacología , Pulpa Dental/citología , Dentina/metabolismo , Regeneración , Células Madre/citología , Animales , Humanos , Inmunofenotipificación , Masculino , Proyectos Piloto , Ratas Wistar , Regeneración/efectos de los fármacos , Células Madre/efectos de los fármacos

RESUMEN

BACKGROUND: The regeneration of dental pulp, especially in cases of pulp death of immature teeth, is the goal of the regenerative endodontic procedures (REPs) that are based on tissue engineering principles, consisting of stem cells, growth factors, and scaffolds. Photobiomodulation therapy (PBMT) showed to improve dental pulp regeneration through cell homing approaches in preclinical studies and has been proposed as the fourth element of tissue engineering. However, when a blood clot was used as a scaffold in one of these previous studies, only 30% of success was achieved. The authors pointed out the instability of the blood clot as the regeneration shortcoming. Then, to circumvent this problem, a new scaffold was developed to be applied with the blood clot. The hypothesis of the present study was that an experimental injectable chitosan hydrogel would facilitate the three-dimensional spatial organization of endogenous stem cells in dental pulp regeneration with no interference on the positive influence of PBMT. METHODS: For the in vitro analysis, stem cells from the apical papilla (SCAPs) were characterized by flow cytometry and applied in the chitosan scaffold for evaluating adhesion, migration, and proliferation. For the in vivo analysis, the chitosan scaffold was applied in a rodent orthotopic dental pulp regeneration model under the influence of PBMT (660 nm; power output of 20 mW, beam area of 0.028 cm2, and energy density of 5 J/cm2). RESULTS: The scaffold tested in this study allowed significantly higher viability, proliferation, and migration of SCAPs in vitro when PBMT was applied, especially with the energy density of 5 J/cm2. These results were in consonance to those of the in vivo data, where pulp-like tissue formation was observed inside the root canal. CONCLUSION: Chitosan hydrogel when applied with a blood clot and PBMT could in the future improve previous results of dental pulp regeneration through cell homing approaches.

Asunto(s)

Quitosano , Pulpa Dental , Terapia por Luz de Baja Intensidad , Regeneración , Andamios del Tejido/química , Animales , Células Cultivadas , Quitosano/química , Quitosano/farmacología , Pulpa Dental/citología , Pulpa Dental/efectos de los fármacos , Pulpa Dental/efectos de la radiación , Humanos , Hidrogeles/química , Masculino , Ratas , Ratas Wistar , Regeneración/efectos de los fármacos , Regeneración/efectos de la radiación , Células Madre/citología , Células Madre/efectos de los fármacos , Células Madre/efectos de la radiación , Ingeniería de Tejidos

RESUMEN

Autologous cell-based therapy for bone regeneration might be impaired by diabetes mellitus (DM) due to the negative effects on mesenchymal stem cells (MSCs) differentiation. Strategies to recover their osteogenic potential could optimize the results. We aimed to evaluate the effect of photobiomodulation (PBM) therapy on osteoblast differentiation of rats with induced DM. Bone marrow MSCs of healthy and diabetic rats were isolated and differentiated into osteoblasts (OB and dOB, respectively). dOB were treated with PBM therapy every 72 hour (660 nm; 0.14 J; 20 mW; 0.714 W/cm2 , and 5 J/cm2 ). Cell morphology, viability, gene and protein expression of osteoblastic markers, alkaline phosphatase (ALP) activity, and the mineralized matrix production of dOB-PBM were compared to dOB. PBM therapy improved viability of dOB, increased the gene and protein expression of bone markers, the ALP activity and the mineralized matrix production. PBM therapy represents an innovative therapeutic approach to optimize the treatment of bone defects in diabetic patients.

Asunto(s)

Diabetes Mellitus Experimental , Terapia por Luz de Baja Intensidad , Células Madre Mesenquimatosas , Animales , Diferenciación Celular , Células Cultivadas , Diabetes Mellitus Experimental/terapia , Humanos , Osteoblastos , Osteogénesis , Ratas

RESUMEN

Photobiomodulation is being widely applied for improving dermal or mucosal wound healing. However, the underlying cellular and molecular processes that directly contribute to its effects remain poorly understood. Pericytes are relevant cells involved in the wound microenvironment and could be one of the main targets of photobiomodulation due to their plasticity and perivascular localization. Herein, we investigate tissue repair under the photobiomodulation stimulus using a pericyte labeled (or reporter) transgenic mice. Using a model of two contralateral back wounds, one the control and the other photoactivated daily (660 nm, 20 mW, 0.71 W/cm2, 5 J/cm2, 7 s, 0.14 J), we showed an overall influx of immune and undifferentiated cells and higher mobilization of a potent pericyte subpopulation (Type-2 pericytes) in the photoactivated wounds in comparison to the controls. Doppler analysis showed a significant increase in the blood flow in the photoactivated wounds, while marked vascular supply was observed histologically. Histochemical analysis has indicated more advanced stages of tissue repair after photoactivation. These data suggest that photobiomodulation significantly accelerates tissue repair through its vascular effects with direct recruitment of pericytes to the injury site.

Asunto(s)

Terapia por Luz de Baja Intensidad , Pericitos/metabolismo , Piel/lesiones , Piel/metabolismo , Cicatrización de Heridas , Animales , Ratones , Ratones Transgénicos , Pericitos/patología , Piel/patología

RESUMEN

Regeneration of diseased bone is challenging. Guided bone regeneration (GBR) has been applied to favor the bone repair. Photobiomodulation (PBM) is also a recognized therapy able to improve bone repair in healthy and diseased individuals. Thus, with the hypothesis that PBM therapy could improve the GBR of diseased bone, this study evaluated the effect of PBM as adjunctive therapy to GBR in osteoporotic rats. Osteoporosis was induced in rats using the oophorectomy model. Then, 5-mm calvaria bone defects were created and treated according to the experimental groups, as follows: with no further treatment (Control); conventional GBR (Membrane), GBR and PBM applied with 3 s, 4 J/cm2 and 0.12 J per point (PBM-1) and GBR and PBM applied with 10s, 14 J/cm2, 0.4 J per point (PBM-2). PBM therapy (808 nm, 40 mW, 1.42 W/cm2) was applied immediately, 48 and 96 h postoperatively. Four and eight weeks later, the samples were harvested and processed for micro-computerized tomography (Micro CT). Data were statistically compared (p < 0.05). From 4 to 8 weeks mostly significant changes were observed in the PBM groups. The bone volume fraction and number of trabeculae of the PBM groups, especially the PBM-1, were significantly higher than those of Control (p < 0.0001). The values of thickness and separation of the trabeculae and structural model index of the PBM groups were significantly smaller than Control (p < 0.0001). The connectivity density was significantly higher on Membrane and PBM groups than Control (p < 0.0004). The application of PBM as adjunctive therapy to GBR results in enhanced bone formation and maturation in comparison to the conventional GBR in the regeneration of lesions of osteoporotic bone in rats. Overviewing the challenges that face bone regeneration in patients with osteoporosis, our findings open new perspectives on the treatment of bone defects under osteoporotic conditions.

Asunto(s)

Regeneración Ósea/efectos de la radiación , Terapia por Luz de Baja Intensidad/métodos , Osteogénesis/efectos de la radiación , Osteoporosis/metabolismo , Cráneo/metabolismo , Animales , Femenino , Rayos Láser , Modelos Animales , Ovariectomía , Ratas , Ratas Wistar , Cráneo/cirugía , Factores de Tiempo , Resultado del Tratamiento , Microtomografía por Rayos X

RESUMEN

OBJECTIVE: The aims of this study were: 1) to evaluate the effect of sintering temperature on microstructure, density and flexural strength of a 3Y-TZP/TiO2 composite containing 12.5 wt% of TiO2 compared to 3Y-TZP specimens (control); 2) to compare 3Y-TZP with the experimental 3Y-TZP/TiO2 composite, both sintered at 1400 °C, with respect to the following parameters: optical properties, characteristic strength, Weibull modulus, fatigue behavior, induction of osteoblasts proliferation and differentiation (mineralization nodules formation). METHODS: The 3Y-TZP and 3Y-TZP/TiO2 powders were uniaxially pressed and sintered at 1200 °C, 1300 °C, 1400 °C or 1500 °C for one hour in a furnace. The microstructural analysis consisted of X-ray diffraction and scanning electron microscopy. The density was measured by the Archimedes' principle and the flexural strength was obtained by the biaxial flexure test. The optical properties were measured using a spectrophotometer operating in the visible light wavelength range. The step-stress accelerated life testing was performed by the pneumatic mechanical cycler and the biological behavior achieved by using osteoblast-like cells (Osteo-1 cell line). RESULTS: Tetragonal zirconia was identified in all groups and cubic zirconia was identified only at 3Y-TZP group. The addition of TiO2 decreased the values of density and flexural strength of the composite 3Y-TZP/TiO2 in relation to 3Y-TZP regardless of the sintering temperature. The color difference between the two materials was not significant regarding L*a*b* parameters. The composite showed higher probability of failure, and induced higher proliferation and differentiation than control. SIGNIFICANCE: The composite developed have good aesthetic and biologics properties. However, its microstructure and mechanical properties need to be improved for future dental implant applications.

Asunto(s)

Materiales Dentales , Itrio , Cerámica , Ensayo de Materiales , Propiedades de Superficie , Temperatura , Titanio , Circonio