Grafting biomaterials associated to topical glucocorticoid: effects on pre-osteoblastic cells (MC3T3-E1).

The topical glucocorticoid budesonide has been prescribed before and after sinus lift surgery as adjuvant drug treatment for maxillary sinus membrane inflammation. However, there is no study on the effects of budesonide on the regenerative process of bone grafting biomaterials. We investigated the effect of the association of budesonide with some biomaterials on the growth and differentiation capacity of pre-osteoblastic cells (MC3T3-E1 subclone 4). Xenogeneic (Bio-Oss and Bio-Gen) and synthetic hydroxyapatites (Osteogen, Bonesynth, and HAP-91) were tested in conditioned medium (1% w/v). The conditioned medium was then supplemented with budesonide (0.5% v/v). Cell viability was assessed using the MTT assay (48, 96, and 144 h), and mineralized nodules were quantified after 14 days of culture using the Alizarin Red Staining. Alkaline phosphatase activity was assessed through the release of thymolphthalein at day seven. All biomaterials showed little or no cytotoxicity. The Bio-Gen allowed significantly less growth than the control group regardless of the experimental time. Regarding differentiation potential of MC3T3-E1, the HAP-91-conditioned medium showed remarkable osteoinductive properties. In osteodifferentiation, the addition of budesonide favored the formation of mineral nodules when cells were cultured in medium conditioned with synthetic materials, whereas it weakened the mineralization potential of cells cultured in xenogeneic medium. Regardless of whether budesonide was added or not, Osteogen and Bio-Oss showed higher alkaline phosphatase activity than the other groups. Budesonide may improve bone formation when associated with synthetic biomaterials. Conversely, the presence of this glucocorticoid weakens the mineralization potential of pre-osteoblastic cells cultured with xenogeneic hydroxyapatites.

Grafting biomaterials associated to topical glucocorticoid: effects on pre-osteoblastic cells (MC3T3-E1)

Abstract The topical glucocorticoid budesonide has been prescribed before and after sinus lift surgery as adjuvant drug treatment for maxillary sinus membrane inflammation. However, there is no study on the effects of budesonide on the regenerative process of bone grafting biomaterials. We investigated the effect of the association of budesonide with some biomaterials on the growth and differentiation capacity of pre-osteoblastic cells (MC3T3-E1 subclone 4). Xenogeneic (Bio-Oss and Bio-Gen) and synthetic hydroxyapatites (Osteogen, Bonesynth, and HAP-91) were tested in conditioned medium (1% w/v). The conditioned medium was then supplemented with budesonide (0.5% v/v). Cell viability was assessed using the MTT assay (48, 96, and 144 h), and mineralized nodules were quantified after 14 days of culture using the Alizarin Red Staining. Alkaline phosphatase activity was assessed through the release of thymolphthalein at day seven. All biomaterials showed little or no cytotoxicity. The Bio-Gen allowed significantly less growth than the control group regardless of the experimental time. Regarding differentiation potential of MC3T3-E1, the HAP-91-conditioned medium showed remarkable osteoinductive properties. In osteodifferentiation, the addition of budesonide favored the formation of mineral nodules when cells were cultured in medium conditioned with synthetic materials, whereas it weakened the mineralization potential of cells cultured in xenogeneic medium. Regardless of whether budesonide was added or not, Osteogen and Bio-Oss showed higher alkaline phosphatase activity than the other groups. Budesonide may improve bone formation when associated with synthetic biomaterials. Conversely, the presence of this glucocorticoid weakens the mineralization potential of pre-osteoblastic cells cultured with xenogeneic hydroxyapatites.

Photobiomodulation drives pericyte mobilization towards skin regeneration.

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.

Evaluation of Resin-Based Material Containing Copaiba Oleoresin (Copaifera Reticulata Ducke): Biological Effects on the Human Dental Pulp Stem Cells.

The purpose of this study was to analyze in vitro the biological effects on human dental pulp stem cells triggered in response to substances leached or dissolved from two experimental cements for dental pulp capping. The experimental materials, based on extracts from Copaifera reticulata Ducke (COP), were compared to calcium hydroxide [Ca(OH)2] and mineral trioxide aggregate (MTA), materials commonly used for direct dental pulp capping in restorative dentistry. For this, human dental pulp stem cells were exposed to COP associated or not with Ca(OH)2 or MTA. Cell cytocompatibility, migration, and differentiation (mineralized nodule formation (Alizarin red assay) and gene expression (RT-qPCR) of OCN, DSPP, and HSP-27 (genes regulated in biomineralization events)) were evaluated. The results showed that the association of COP reduced the cytotoxicity of Ca(OH)2. Upregulations of the OCN, DSPP, and HSP-27 genes were observed in response to the association of COP to MTA, and the DSPP and HSP-27 genes were upregulated in the Ca(OH)2 + COP group. In up to 24 h, cell migration was significantly enhanced in the MTA + COP and Ca(OH)2 + COP groups. In conclusion, the combination of COP with the currently used materials for dental pulp capping [Ca(OH)2 and MTA] improved the cell activities related to pulp repair (i.e., cytocompatibility, differentiation, mineralization, and migration) including a protective effect against the cytotoxicity of Ca(OH)2.

Photobiomodulation effects on keratinocytes cultured in vitro: a critical review.

Photobiomodulation therapy (PBMT) has been widely used for the promotion of tissue repair. Despite these therapeutic benefits, in some cases, PBMT appears to be unsuccessful, and the strongest hypothesis is that this failure is due to inadequate light dosimetry and wavelengths. The objective of the present critical review was to evaluate the effects of PBMT on cultured keratinocytes using blue, red, or near-infrared light categorized into arbitrary ranges of energy density (0.1-5.0, 5.1-10.0, 10.1-15.0, and over 15.0 J/cm2). The electronic searches were conducted in PubMed, Web of Science, Scopus and LILACS databases, and included LASER or LED devices. A total of 55 articles evaluating the effects of PBMT on cell viability, proliferation, migration, and cytokine and growth factor production were included. Overall, the studies failed to provide detailed information about light dosimetry or detailed experimental conditions. The vast majority of the energy densities tested produced unmodified results regardless of the wavelength applied. However, it was possible to observe that red and near-infrared light had more stimulatory effects than blue light. In addition, for all parameters analyzed, favorable outcomes were mostly obtained in the range of 0.1-5.0 J/cm2. The less explored energy densities were within the 10.1-15.0 J/cm2 range. Energy densities above 15.0 J/cm2 were ineffective or tended to cause cell death. The heterogeneity of the data does not allow us to define a PBMT range setting protocol that would have beneficial effects on keratinocytes.

Bioactive glass plus laser phototherapy as promise candidates for dentine hypersensitivity treatment.

Treatments for dentine hypersensitivity (DH) may produce positive effects, though do not have lasting results. We investigated the reparative potential of stem cells derived from deciduous teeth (SHEDs) in response to components delivered from substances used in the treatment of the DH, associated or not to laser phototherapy (LPT), to stimulate dentine formation. SHEDs were submitted to substances delivered from a laboratorial P-rich bioactive glass [57SiO2 -26CaO-17P2 O5 (wt %)] or a commercially available desensitizer (Gluma® Desensitizer), associated (or not) to LPT (InGAlP diode laser, 660 nm, 0.028 cm2 , 20 mW, 5 J/cm2 , 7 s, contact mode). Biomaterial characterization was performed by X-ray diffraction, scanning electron microscopy and the particle size was evaluated by dynamic light scattering. SHEDs proliferation and differentiation were analyzed by MTT and Alizarin Red staining, respectively. The conditioned media used in these tests were evaluated regarding their pH and the ionic concentration changes due to ions leached from the bioactive glass (BG). BG majority presented a non-crystalline solid structure and mixed particle sizes characterized by the agglomeration of nanoparticles. Cultures treated with BG alone or in association to LPT showed improved cell growth in relation to Gluma® (p < 0.05). Gluma® was cytotoxic in all tested conditions, regardless irradiated or not. BG associated to LPT induced intense mineral matrix formation. In conclusion, BG releases ionic dissolution products able to promote SHEDs differentiation. BG associated to LPT improves SHEDs proliferation and differentiation in vitro, and may be a promise therapeutic approach for the DH treatment. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 107-116, 2017.

Photobiomodulation of Dental Derived Mesenchymal Stem Cells: A Systematic Review.

OBJECTIVE: This study aimed to conduct a systematic review of the literature published from 2000 to August 2015, to investigate the effect of photobiomodulation (PBM) therapy on dentoalveolar-derived mesenchymal stem cells (ddMSCs), assessing whether a clear conclusion can be reached from the data presented. BACKGROUND DATA: Systematic reviews provide the best evidence on the effectiveness of a procedure and permit investigation of factors that may influence the performance of a method. To the best of our knowledge, no previous systematic review has evaluated the effects of PBM only on ddMSCs. METHODS: The search was conducted in PubMed /MEDLINE®, Scopus and Web of Science databases, and reported according to the Preferred Reporting Items for Systematic Reviews and Metaanalyses (PRISMA Statement). Original research articles investigating the effects of PBM therapy on ddMSCs, published from 2000 to August 2015, were retrieved and used for this review according to the following eligibility criteria: evaluating PBM therapy, assessing stem cells of dentoalveolar origin, published in English, dealing with cells characterized as stem cells, and using light that did not need external chromophores. RESULTS: From the initial 3467 potentially relevant articles identified, 6 were excluded because they were duplicates, and 3453 were considered ineligible based on the inclusion criteria. Therefore, eight articles remained, and these were fully analyzed in order to closely check exclusion criteria items. Only one of them was excluded because the cultured cells studied were not characterized as stem cells. Finally, seven articles served as the basis for this systematic review. CONCLUSIONS: PBM therapy has no deleterious effects on ddMSCs. Although no other clear conclusion was obtained because of the scarce number of publications, the results of these studies are pointing to an important tendency of PBM therapy to improve ddMSCs' viability and proliferation.

Laser phototherapy enhances mesenchymal stem cells survival in response to the dental adhesives.

BACKGROUND: We investigated the influence of laser phototherapy (LPT) on the survival of human mesenchymal stem cells (MSCs) submitted to substances leached from dental adhesives. METHOD: MSCs were isolated and characterized. Oral mucosa fibroblasts and osteoblast-like cells were used as comparative controls. Cultured medium conditioned with two adhesive systems was applied to the cultures. Cell monolayers were exposed or not to LPT. Laser irradiations were performed using a red laser (GaAlAs, 780 nm, 0.04 cm(2), 40 mW, 1 W/cm(2), 0.4 J, 10 seconds, 1 point, 10 J/cm(2)). After 24 h, cell viability was assessed by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide reduction assay. Data were statistically compared by ANOVA followed by Tukey's test (P < 0.05). RESULTS: Different cell types showed different viabilities in response to the same materials. Substances leached from adhesives were less cytotoxic to MSCs than to other cell types. Substances leached from Clearfil SE Bond were highly cytotoxic to all cell types tested, except to the MSCs when applied polymerized and in association with LPT. LPT was unable to significantly increase the cell viability of fibroblasts and osteoblast-like cells submitted to the dental adhesives. CONCLUSION: LPT enhances mesenchymal stem cells survival in response to substances leached from dental adhesives.

Diferenciação odonto/osteogênica de células-tronco mesenquimais fotomoduladas em hidrogel com incorporação de proteína morfogenética óssea 4 / Odonto/osteogenic differentiation of photomodulated mesenchymal stem cells in BMP4-loaded hydrogel

Este estudo avaliou a influência da fototerapia a laser (FTL) na proliferação e diferenciação de células-tronco da polpa dentária humana (DPSCs; do inglês, Dental Pulp Stem Cells ) encapsuladas em carreador injetável e termoresponsivo (PL; Pluronic® F-127, Sigma-Aldrich, MO, EUA) com incorporação de proteína morfogenética óssea 4 recombinante humana (rhBMP4) (sistema PL/rhBMP4). O biomaterial foi caracterizado de acordo com seus perfis de embebição e dissolução, liberação de rhBMP4 e sua estrutura morfológica. DPSCs foram isoladas, caracterizadas e encapsuladas em PL para confirmar sua viabilidade e seu potencial de diferenciação (adipo e osteogênico) em comparação com células-tronco mesenquimais de medula óssea (BMMSCs; do inglês, Bone Marrow Mesenchymal Stem Cells). Quando encapsuladas no sistema PL/rhBMP4, DPSCs foram irradiadas com duas densidades de energia diferentes utilizando laser de diodo de fosfeto de índio-gálio-alumínio (InGaAlP), modos contínuo, pontual e em contato [660 nm, 0,028 cm2, 20 mW, 0,71 W/cm2, 3 J/cm2 (4 s) ou 5 J/cm2 (7 s)]. Os ensaios de PKH26 (do inglês, Red Fluorescent Cell Linker), CFU-F (do inglês, Coloning Forming Units - Fibroblastic), e MTT (do inglês, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide)) foram utilizados para avaliar adesão/proliferação, diferenças na capacidade formadora de colônias e viabilidade das DPSCs (neste último caso sob estresse nutricional), respectivamente. Finalmente, a diferenciação odonto/osteogênica foi analisada por qRT-PCR e confirmada por ensaio de vermelho de alizarina. O biomaterial embebeu e dissolveu rapidamente; densa rede tubular e reticular com poros interconectados foi observada. DPSCs e BMMSCs apresentaram alta viabilidade celular quando encapsuladas em PL. Ambas as linhagens celulares tiveram êxito em se diferenciar em tecidos adiposo e ósseo. De acordo com o PKH26, DPSCs puderam aderir e proliferar no sistema PL/rhBMP4. DPSCs irradiadas encapsuladas tanto em PL como em PL/rhBMP4 formaram mais CFU-F que os controles não irradiados. Sob estresse nutricional, DPSCs semeadas no PL e irradiadas com 5 J/cm2 exibiram maior taxa de viabilidade celular em relação aos grupos não irradiados e irradiados com 3 J/cm2. Na presença de rhBMP4, os grupos irradiados tanto com 3 J/cm2 quanto com 5 J/cm2 apresentaram deposição mineral precoce quando comparados aos grupos não irradiados. Ainda, após 21 dias de diferenciação odonto/osteogênica, DPSCs irradiadas produziram maior quantidade de nódulos mineralizados. A irradiação com 5 J/cm2 levou ao aumento significativo da expressão de genes envolvidos na diferenciação odonto/osteogênica, como colágeno tipo I (COL1A1), osteocalcina (OCN), proteína da matriz dentinária 1 (DMP1), sialofosfoproteina dentinária (DSPP) e proteína heat shock 27 kDa (HSPB1). A associação entre rhBMP4 e FTL promove proliferação e diferenciação odonto/osteogênica de DPSCs acelerando e aumentando notavelmente a formação de tecido mineralizado, em especial quando a densidade de energia de 5 J/cm2 é aplicada.

This study evaluated the influence of laser phototherapy (LPT) on dental pulp stem cells (DPSCs) proliferation and differentiation upon encapsulation in an injectable and thermo-responsive cell carrier (PL; Pluronic® F-127, Sigma-Aldrich, MO, USA) loaded with human recombinant bone morphogenetic protein 4 (rhBMP4)(PL/rhBMP4 system). The biomaterial was characterized according to its swelling and dissolution profiles, release of rhBMP4 and morphological structure. DPSCs were isolated, characterized and encapsulated in PL to confirm their viability and multilineage differentiation potential (adipo and osteogenic) in comparison to bone marrow mesenchymal stem cells (BMMSCs). When encapsulated in the PL/rhBMP4 system, DPSCs were irradiated with two different energy densities using a continuous-wave indium-gallium-aluminum-phosphide (InGaAlP) diode laser [660 nm, 0.028 cm2, 20 mW, 0.71 W/cm2, 3 J/cm2 (4 s) or 5 J/cm2 (7 s)] in punctual and contact modes. The PKH26 (Red Fluorescent Cell Linker), the CFU-F (Coloning Forming Units - Fibroblastic), and the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] assays were used to assess differences in cell adhesion/proliferation, colony forming units formation ability, and cell viability of DPSCs (in this case under nutritional stress), respectively. Then, alizarin red and qRT-PCR analyzes were used to evaluate odonto/osteogenic differentiation. The biomaterial swelled and dissolved rapidly; dense tubular and reticular network morphology with well-interconnected pores was observed. DPSCs and BMMSCs presented high cell viability when encapsulated in PL. Both cell lineages successfully differentiated into bone or adipose tissues. According to PKH26, DPSCs were able to adhere and proliferate in the PL/rhBMP4 system. Irradiated DPSCs encapsulated in either PL or PL/rhBMP4 system formed more CFU-F than non-irradiated controls. Under nutritional stress, DPSCs encapsulated in the hydrogels with no rhBMP4 and irradiated at 5 J/cm2 exhibited higher cell viability than the other groups. In the presence of rhBMP4, the groups irradiated both at 3 and 5 J/cm2 energy densities displayed earlier mineral deposition than the non-irradiated groups. Moreover, after 21 days of odonto/osteogenic differentiation, irradiated DPSCs produced greater nodule formation than the control groups. At the energy density of 5 J/cm2, there were significant upregulation of genes involved in odonto/osteoblast differentiation, such as type I collagen (COL1A1), osteocalcin (OCN), dentin matrix protein 1 (DMP1), dentin sialophosphoprotein (DSPP) and heat shock protein 27 kDa (HSPB1). The association between rhBMP4 and LPT promotes cell proliferation and odonto/osteogenic differentiation of DPSCs accelerating and increasing the formation of mineralized tissue, in particular when the energy density of 5 J/cm2 is applied.

Diferenciação odonto/osteogênica de células-tronco mesenquimais fotomoduladas em hidrogel com incorporação de proteína morfogenética óssea 4 / Odonto/osteogenic differentiation of photomodulated mesenchymal stem cells in BMP4-loaded hydrogel

Este estudo avaliou a influência da fototerapia a laser (FTL) na proliferação e diferenciação de células-tronco da polpa dentária humana (DPSCs; do inglês, Dental Pulp Stem Cells ) encapsuladas em carreador injetável e termoresponsivo (PL; Pluronic® F-127, Sigma-Aldrich, MO, EUA) com incorporação de proteína morfogenética óssea 4 recombinante humana (rhBMP4) (sistema PL/rhBMP4). O biomaterial foi caracterizado de acordo com seus perfis de embebição e dissolução, liberação de rhBMP4 e sua estrutura morfológica. DPSCs foram isoladas, caracterizadas e encapsuladas em PL para confirmar sua viabilidade e seu potencial de diferenciação (adipo e osteogênico) em comparação com células-tronco mesenquimais de medula óssea (BMMSCs; do inglês, Bone Marrow Mesenchymal Stem Cells). Quando encapsuladas no sistema PL/rhBMP4, DPSCs foram irradiadas com duas densidades de energia diferentes utilizando laser de diodo de fosfeto de índio-gálio-alumínio (InGaAlP), modos contínuo, pontual e em contato [660 nm, 0,028 cm2, 20 mW, 0,71 W/cm2, 3 J/cm2 (4 s) ou 5 J/cm2 (7 s)]. Os ensaios de PKH26 (do inglês, Red Fluorescent Cell Linker), CFU-F (do inglês, Coloning Forming Units - Fibroblastic), e MTT (do inglês, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide)) foram utilizados para avaliar adesão/proliferação, diferenças na capacidade formadora de colônias e viabilidade das DPSCs (neste último caso sob estresse nutricional), respectivamente. Finalmente, a diferenciação odonto/osteogênica foi analisada por qRT-PCR e confirmada por ensaio de vermelho de alizarina. O biomaterial embebeu e dissolveu rapidamente; densa rede tubular e reticular com poros interconectados foi observada.

DPSCs e BMMSCs apresentaram alta viabilidade celular quando encapsuladas em PL. Ambas as linhagens celulares tiveram êxito em se diferenciar em tecidos adiposo e ósseo. De acordo com o PKH26, DPSCs puderam aderir e proliferar no sistema PL/rhBMP4. DPSCs irradiadas encapsuladas tanto em PL como em PL/rhBMP4 formaram mais CFU-F que os controles não irradiados. Sob estresse nutricional, DPSCs semeadas no PL e irradiadas com 5 J/cm2 exibiram maior taxa de viabilidade celular em relação aos grupos não irradiados e irradiados com 3 J/cm2. Na presença de rhBMP4, os grupos irradiados tanto com 3 J/cm2 quanto com 5 J/cm2 apresentaram deposição mineral precoce quando comparados aos grupos não irradiados. Ainda, após 21 dias de diferenciação odonto/osteogênica, DPSCs irradiadas produziram maior quantidade de nódulos mineralizados. A irradiação com 5 J/cm2 levou ao aumento significativo da expressão de genes envolvidos na diferenciação odonto/osteogênica, como colágeno tipo I (COL1A1), osteocalcina (OCN), proteína da matriz dentinária 1 (DMP1), sialofosfoproteina dentinária (DSPP) e proteína heat shock 27 kDa (HSPB1). A associação entre rhBMP4 e FTL promove proliferação e diferenciação odonto/osteogênica de DPSCs acelerando e aumentando notavelmente a formação de tecido mineralizado, em especial quando a densidade de energia de 5 J/cm2 é aplicada.

This study evaluated the influence of laser phototherapy (LPT) on dental pulp stem cells (DPSCs) proliferation and differentiation upon encapsulation in an injectable and thermo-responsive cell carrier (PL; Pluronic® F-127, Sigma-Aldrich, MO, USA) loaded with human recombinant bone morphogenetic protein 4 (rhBMP4)(PL/rhBMP4 system). The biomaterial was characterized according to its swelling and dissolution profiles, release of rhBMP4 and morphological structure. DPSCs were isolated, characterized and encapsulated in PL to confirm their viability and multilineage differentiation potential (adipo and osteogenic) in comparison to bone marrow mesenchymal stem cells (BMMSCs). When encapsulated in the PL/rhBMP4 system, DPSCs were irradiated with two different energy densities using a continuous-wave indium-gallium-aluminum-phosphide (InGaAlP) diode laser [660 nm, 0.028 cm2, 20 mW, 0.71 W/cm2, 3 J/cm2 (4 s) or 5 J/cm2 (7 s)] in punctual and contact modes. The PKH26 (Red Fluorescent Cell Linker), the CFU-F (Coloning Forming Units - Fibroblastic), and the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] assays were used to assess differences in cell adhesion/proliferation, colony forming units formation ability, and cell viability of DPSCs (in this case under nutritional stress), respectively. Then, alizarin red and qRT-PCR analyzes were used to evaluate odonto/osteogenic differentiation. The biomaterial swelled and dissolved rapidly; dense tubular and reticular network morphology with well-interconnected pores was observed. DPSCs and BMMSCs presented high cell viability when encapsulated in PL. Both cell lineages successfully differentiated into bone or adipose tissues. According to PKH26,

DPSCs were able to adhere and proliferate in the PL/rhBMP4 system. Irradiated DPSCs encapsulated in either PL or PL/rhBMP4 system formed more CFU-F than non-irradiated controls. Under nutritional stress, DPSCs encapsulated in the hydrogels with no rhBMP4 and irradiated at 5 J/cm2 exhibited higher cell viability than the other groups. In the presence of rhBMP4, the groups irradiated both at 3 and 5 J/cm2 energy densities displayed earlier mineral deposition than the non-irradiated groups. Moreover, after 21 days of odonto/osteogenic differentiation, irradiated DPSCs produced greater nodule formation than the control groups. At the energy density of 5 J/cm2, there were significant upregulation of genes involved in odonto/osteoblast differentiation, such as type I collagen (COL1A1), osteocalcin (OCN), dentin matrix protein 1 (DMP1), dentin sialophosphoprotein (DSPP) and heat shock protein 27 kDa (HSPB1). The association between rhBMP4 and LPT promotes cell proliferation and odonto/osteogenic differentiation of DPSCs accelerating and increasing the formation of mineralized tissue, in particular when the energy density of 5 J/cm2 is applied.