The Effects In Vitro of Photobiomodulation Over Fibroblasts and Extracellular Matrix.

Objective: The objective of this study is to evaluate the potential effects of photobiomodulation (PBM) on cell proliferation and extracellular matrix production of human fibroblasts (FN1) cultured in 2D. Background: Patients with healing difficulties suffer injuries that take time to recover. In addition, aging can be seen in our faces daily when we look in the mirror; in both situations, collagen production is reduced. Fibroblasts act in the beginning and at the end of the inflammation phase, signaling to immune agents, and platelets, and producing collagen, coordinating repair. PBM increases cell viability, proliferation, and mRNA production. Methods: Human fibroblasts were irradiated three times after cell seed (after 24, 48, and 72 h) using a gallium-aluminum arsenideGaAlAs low-level laser (LLL). Cell viability, proliferative response, synthesis of collagen types I and III, and soluble collagen production were analyzed. The statistical significance of differences between groups was determined using unpaired one-way analysis of variance (ANOVA) p < 0.05. Results: PBM increased significantly the number of fibroblasts, and the production of collagen types I (Col I) and III (Col III), after three sessions of LLL with 2.5 J per session, every 24 h, for 3 consecutive days; total energy delivered after 72 h is 7.5 J. Conclusions: This energy density of LLL increases fibroblast proliferation and collagen production in vitro without side effects.

Three-Dimensional Culture Systems for Dissecting Notch Signalling in Health and Disease.

Three-dimensional (3D) culture systems opened up new horizons in studying the biology of tissues and organs, modelling various diseases, and screening drugs. Producing accurate in vitro models increases the possibilities for studying molecular control of cell-cell and cell-microenvironment interactions in detail. The Notch signalling is linked to cell fate determination, tissue definition, and maintenance in both physiological and pathological conditions. Hence, 3D cultures provide new accessible platforms for studying activation and modulation of the Notch pathway. In this review, we provide an overview of the recent advances in different 3D culture systems, including spheroids, organoids, and "organ-on-a-chip" models, and their use in analysing the crucial role of Notch signalling in the maintenance of tissue homeostasis, pathology, and regeneration.

Bovine pericardium membrane, gingival stem cells, and ascorbic acid: a novel team in regenerative medicine.

Recently, the development and the application of 3D scaffold able to promote stem cell differentiation represented an essential field of interest in regenerative medicine. In particular, functionalized scaffolds improve bone tissue formation and promote bone defects repair. This research aims to evaluate the role of ascorbic acid (AS) supplementation in an in vitro model, in which a novel 3D-scaffold, bovine pericardium collagen membrane called BioRipar (BioR) was functionalized with human Gingival Mesenchymal Stem Cells (hGMSCs). As extensively reported in the literature, AS is an essential antioxidant molecule involved in the extracellular matrix secretion and in the osteogenic induction. Specifically, hGMSCs were seeded on BioR and treated with 60 and 90 µg/mL of AS in order to assess their growth behavior, the expression of bone specific markers involved in osteogenesis (runt-related transcription factor 2, RUNX2; collagen1A1, COL1A1; osteopontin, OPN; bone morphogenetic protein2/4, BMP2/4), and de novo deposition of calcium. The expression of COL1A1, RUNX2, BMP2/4 and OPN was evaluated by RT-PCR, Western blotting and immunocytochemistry, and proved to be upregulated. Our results demonstrate that after three weeks of treatment AS at 60 and 90 µg/mL operates as an osteogenic inductor in hGMSCs. These data indicate that the AS supplementation produces an enhancement of osteogenic phenotype commitment in an in vitro environment. For this reason, AS could represent a valid support for basic and translational research in tissue engineering and regenerative medicine.

Laser Photobiomodulation Over Teeth Subjected to Orthodontic Movement.

Background: Orthodontics of the 21st century requires aesthetic, painless, predictable, and quick treatments. This demand for faster results generated orthodontic movement acceleration protocols (OMAPs); among other OMAPs we present low-level laser (LLL) as a candidate. Objective: To evaluate levels of interleukin (IL)-1, IL-10, and type 1 collagen in the periodontal ligament of first molars of rats subjected to orthodontic traction with and without LLL irradiation, compared with untreated controls (CO), and to evaluate whether the dose of LLL used in this work is eligible as an OMAP. Materials and methods: A total of 35 male Wistar rats were distributed into three groups: group 1 NI (nonirradiated) n = 15, group 2 IR (laser irradiated using 5 J, 177 J/cm2, and 100 mW applied in contact to the vestibular mesial, vestibular distal, and palatal faces of gum tissue around molar region for 50 sec each point, for 3 consecutive days, immediately 24 and 48 h after orthodontic device placement.) n = 15, and group 3 CO n = 5; groups 1 and 2 were subjected to orthodontic force and each group was divided into three subgroups that were sacrificed after 3, 5, and 7 days, IL-1/10 and COL-1 levels were analyzed. Results: In the IR group, levels of IL-1/10 and COL-1 showed peak anticipation after LLL irradiation compared with those in the NI and CO groups. Conclusions: These results can also infer that this dose of LLL can be used as an OMAP.

Laser photobiomodulation in pressure ulcer healing of human diabetic patients: gene expression analysis of inflammatory biochemical markers.

Pressure ulcers (PU) are wounds located mainly on bone surfaces where the tissue under pressure suffers ischemia leading to cellular lesion and necrosis , its causes and the healing process depend on several factors. The aim of this study was evaluating the gene expression of inflammatory/reparative factors: IL6, TNF, VEGF, and TGF, which take part in the tissue healing process under effects of low-level laser therapy (LLLT). In order to perform lesion area analysis, PUs were photographed and computer analyzed. Biochemical analysis was performed sa.mpling ulcer border tissue obtained through biopsy before and after laser therapy and quantitative real-time PCR (qRT-PCR) analysis. The study comprised eight individuals, mean age sixty-two years old, and sacroiliac and calcaneous PU, classified as degree III and IV according to the National Pressure Ulcer Advisory Panel (NPUAP). PUs were irradiated with low-level laser (InGaAIP, 100 mW, 660 nm), energy density 2 J/cm2, once a day, with intervals of 24 h, totaling 12 applications. The lesion area analysis revealed averaged improvement of the granulation tissue size up to 50% from pre- to post-treatment. qRT-PCR analysis revealed that IL6 values were not significantly different before and after treatment, TNF gene expression was reduced, and VEFG and TGF-ß gene expression increased after treatment. After LLLT, wounds presented improvement in gross appearance, with increase in factors VEFG and TGF-ß, and reduction of TNF; despite our promising results, they have to be analyzed carefully as this study did not have a control group.

Comparative Study of the Physiotherapeutic and Drug Protocol and Low-Level Laser Irradiation in the Treatment of Pain Associated with Temporomandibular Dysfunction.

BACKGROUND: The temporomandibular joint (TMJ) is a structure of the craniofacial complex affected by neurological diseases. Orthopedic and musculoskeletal changes can also cause temporomandibular disorders (TMD) and pain. Low-level laser (LLL) therapy has been studied in the treatment of temporomandibular jaw (TMJ) dysfunction, and controversial results were obtained. OBJECTIVE: The objective of this work was comparing the physiotherapeutic and drug protocol (PDP) to LLL therapy in the treatment of pain associated with TMD. METHODS: A sample of 60 female patients, 20-50 years of age, TMD triggering agents (stress, parafunctional habits) controlled, was randomly divided into three groups, group 1 (G1)-LLL (780 nm laser, dose of 35.0 J/cm2, for 20 sec, thrice a week, for 4 weeks); group 2 (G2)-PDP (hot packs thrice a day, morning, afternoon, and evening, for 15 min, exercise of opening and closing the mouth, twice a day, myorelaxing and anti-inflammatory drug administration); and group 3 (G3)-Placebo (450 nm halogen lamp, Max LD Gnatus, light curing unit). RESULTS: Patients were evaluated every return appointment for the presence (P) or absence (A) of pain for 4 weeks and results were statistically analyzed. First week: 60% of G1, 100% G2, and 70% of G3-related pain. Second week: 55% of G1, 15% of G2, and 100% of G3-related pain. Third week: 10% of G1, 15% of G2, and 85% of G3-related pain. Last week: 0% of G1, 0% of G2, and 100% of G3-related pain. CONCLUSIONS: Based on obtained data, we concluded that, compared to PDP, LLL treatment is effective to control pain associated with TMD.

Evaluation of the Proliferative Effects Induced by Low-Level Laser Therapy in Bone Marrow Stem Cell Culture.

OBJECTIVE: The objective of this study was to evaluate the effect of laser irradiation on dog bone marrow stem cells. BACKGROUND DATA: Low doses of low-level red laser positively affect the viability of mesenchymal stem cells, and also increase proliferation. METHODS: Low-level laser (wavelength, 660 nm; power output, 50 mW), was applied to dog bone marrow stem cell cultures (DBMSC). The energy densities delivered varied from 1 to 12J/cm(2). The effect of the laser irradiation was evaluated on cell proliferation measured with the MTT colorimetric test, cell cycle phase, and on lipidic peroxidation (free radical production). RESULTS: The results indicate that laser irradiation to DBMSC did not change the morphology of the cells, but significantly increased their viability and the number of cells at the G2/M phase with 6, 10, and 12 J/cm(2). On the other hand, malonaldehyde production was significantly enhanced with 8 J/cm(2). CONCLUSIONS: The parameters used to irradiate DBMSC increased significantly proliferation without producing high levels of reactive oxygen species (ROS).