RESUMEN
Osteoporosis and other degenerative bone diseases pose significant challenges to global healthcare systems due to their prevalence and impact on quality of life. Current treatments often alleviate symptoms without fully restoring damaged bone tissue, highlighting the need for innovative approaches like stem cell therapy. Adipose-derived mesenchymal stem cells (ADMSCs) are particularly promising due to their accessibility, abundant supply, and strong differentiation potential. However, ADMSCs tend to favor adipogenic pathways, necessitating the use of differentiation inducers (DIs), three-dimensional (3D) hydrogel environments, and photobiomodulation (PBM) to achieve targeted osteogenic differentiation. This study investigated the combined effects of osteogenic DIs, a fast-dextran hydrogel matrix, and PBM at specific wavelengths and fluences on the proliferation and differentiation of immortalized ADMSCs into osteoblasts. Near-infrared (NIR) and green (G) light, as well as their combination, were used with fluences of 3 J/cm2, 5 J/cm2, and 7 J/cm2. The results showed statistically significant increases in alkaline phosphatase levels, a marker of osteogenic differentiation, with G light at 7 J/cm2 demonstrating the most substantial impact on ADMSC differentiation. Calcium deposits, visualized by Alizarin red S staining, appeared as early as 24 h post-treatment in PBM groups, suggesting accelerated osteogenic differentiation. ATP luminescence assays indicated increased proliferation in all experimental groups, particularly with NIR and NIR-G light at 3 J/cm2 and 5 J/cm2. MTT viability and LDH membrane permeability assays confirmed enhanced cell viability and stable cell health, respectively. In conclusion, PBM significantly influences the differentiation and proliferation of hydrogel-embedded immortalized ADMSCs into osteoblast-like cells, with G light at 7 J/cm2 being particularly effective. These findings support the combined use of 3D hydrogel matrices and PBM as a promising approach in regenerative medicine, potentially leading to innovative treatments for degenerative bone diseases.
Asunto(s)
Diferenciación Celular , Terapia por Luz de Baja Intensidad , Células Madre Mesenquimatosas , Osteogénesis , Osteogénesis/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Células Madre Mesenquimatosas/efectos de la radiación , Humanos , Terapia por Luz de Baja Intensidad/métodos , Técnicas de Cultivo Tridimensional de Células/métodos , Proliferación Celular/efectos de la radiación , Tejido Adiposo/citología , Hidrogeles/química , Osteoblastos/citología , Osteoblastos/metabolismo , Osteoblastos/efectos de la radiación , Fosfatasa Alcalina/metabolismo , Células CultivadasRESUMEN
This study aimed to compare the effects of photobiomodulation therapy (PBMT) with 660 and 980 nm diode lasers on differentiation of periodontal ligament mesenchymal stem cells (PDLMSCs). In this in vitro, experimental study, PDLMSCs were obtained from the Iranian Genetic Bank and cultured in osteogenic medium. They were then subjected to irradiation of 660 and 980 nm diode lasers, and their viability was assessed after one, two, and three irradiation cycles using the methyl thiazolyl tetrazolium (MTT) assay. The cells also underwent DAPI staining, cell apoptosis assay by using the Annexin V/PI, Alizarin Red staining, and real-time polymerase chain reaction (PCR) for assessment of the expression of osteogenic genes. Data were analyzed by two-way ANOVA. The two laser groups had no significant difference in cell apoptosis according to the results of DAPI staining. Both laser groups showed higher cell viability in the MTT assay at 4 and 6 days compared with the control group. Annexin V/PI results showed higher cell viability in both laser groups at 4 days compared with the control group. Rate of early and late apoptosis was lower in both laser groups than the control group at 4 days. Necrosis had a lower frequency in 980 nm laser group than the control group on day 6. Alizarin Red staining showed higher cell differentiation in both laser groups after 3 irradiation cycles than the control group. The highest expression of osteopontin (OPN), osteocalcin (OCN), and Runt-related transcription factor 2 (RUNX2) was noted in 660 nm laser group with 3 irradiation cycles at 14 days, compared with the control group. PBMT with 660 and 980 nm diode lasers decreased apoptosis and significantly increased PDLMSC differentiation after 3 irradiation cycles.
Asunto(s)
Apoptosis , Diferenciación Celular , Supervivencia Celular , Láseres de Semiconductores , Terapia por Luz de Baja Intensidad , Células Madre Mesenquimatosas , Osteogénesis , Ligamento Periodontal , Ligamento Periodontal/efectos de la radiación , Ligamento Periodontal/citología , Células Madre Mesenquimatosas/efectos de la radiación , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Diferenciación Celular/efectos de la radiación , Humanos , Terapia por Luz de Baja Intensidad/métodos , Láseres de Semiconductores/uso terapéutico , Supervivencia Celular/efectos de la radiación , Apoptosis/efectos de la radiación , Osteogénesis/efectos de la radiación , Células Cultivadas , Osteocalcina/metabolismo , Osteocalcina/genética , Osteopontina/metabolismo , Osteopontina/genética , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genéticaRESUMEN
Introduction: In the process of bone regeneration, a prominent role is played by macrophages involved in both the initial inflammation and the regeneration/vascularization phases, due to their M2 anti-inflammatory phenotype. Together with osteoclasts, they participate in the degradation of the bone matrix if the inflammatory process does not end. In this complex scenario, recently, much attention has been paid to extracellular communication mediated by nanometer-sized vesicles, with high information content, called exosomes (EVs). Considering these considerations, the purpose of the present work is to demonstrate how the presence of a pulsed electromagnetic field (PEMF) can positively affect communication through EVs. Methods: To this aim, macrophages and osteoclasts were treated in vitro with PEMF and analyzed through molecular biology analysis and by electron microscopy. Moreover, EVs produced by macrophages were characterized and used to verify their activity onto osteoclasts. Results: The results confirmed that PEMF not only reduces the inflammatory activity of macrophages and the degradative activity of osteoclasts but that the EVS produced by macrophages, obtained from PEMF treatment, positively affect osteoclasts by reducing their activity. Discussion: The co-treatment of PEMF with M2 macrophage-derived EVs (M2-EVs) decreased osteoclastogenesis to a greater degree than separate treatments.
Asunto(s)
Regeneración Ósea , Campos Electromagnéticos , Exosomas , Macrófagos , Osteoclastos , Osteogénesis , Exosomas/química , Exosomas/metabolismo , Regeneración Ósea/efectos de la radiación , Regeneración Ósea/fisiología , Animales , Ratones , Osteogénesis/fisiología , Osteogénesis/efectos de la radiación , Células RAW 264.7RESUMEN
BACKGROUND/AIM: Bone marrow cells contain nonhematopoietic cells with the ability to differentiate into osteogenic, chondrogenic, and adipogenic lineages. Mechanical stress influences osteoblast differentiation of bone marrow cells into osteogenic, chondrogenic, and adipogenic lineages, measurable as the abundance of alkaline phosphatase-positive (ALP+) colony-forming unit-fibroblasts (CFU-F); however, the effect of diode laser irradiation on osteoblast differentiation is unknown. The aim of this study was to analyze the effects of photobiomodulation on the osteogenic differentiation of mesenchymal stem cells in the bone marrow, using the CFU-F assay. MATERIALS AND METHODS: Bone marrow cells isolated from rat tibiae were cultured and irradiated with a diode laser (wavelength 808 nm) at a total energy of 0 J (control), 50 J, and 150 J. RESULTS: On day 7 after irradiation, ALP+ CFU-F were most abundant in the 50 J group and the least abundant in the 150 J group. Mineralized nodule formation was observed after long-term culture (21 days). Compared with the control group, there were significantly more nodules in the 50 J group and significantly fewer nodules in the 150 J group. Osteocalcin mRNA expression was highest in the 50 J group, and there was no difference between the control and 150 J groups. CONCLUSION: Irradiation with 50 J was effective in stimulating osteogenesis in bone marrow stem cells. These findings suggest that diode laser irradiation can induce osteogenesis in rat bone marrow cells in an energy-dependent manner, and appears suitable for application in bone regeneration therapy.
Asunto(s)
Células de la Médula Ósea , Diferenciación Celular , Láseres de Semiconductores , Células Madre Mesenquimatosas , Osteoblastos , Osteogénesis , Animales , Diferenciación Celular/efectos de la radiación , Ratas , Osteogénesis/efectos de la radiación , Células de la Médula Ósea/efectos de la radiación , Células de la Médula Ósea/citología , Células Madre Mesenquimatosas/efectos de la radiación , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Osteoblastos/efectos de la radiación , Osteoblastos/citología , Osteoblastos/metabolismo , Células Cultivadas , Fosfatasa Alcalina/metabolismo , Fosfatasa Alcalina/genética , Masculino , Terapia por Luz de Baja Intensidad/métodos , Osteocalcina/metabolismo , Osteocalcina/genéticaRESUMEN
The present study aimed to evaluate the effect of photobiomodulation therapy (PBM) on different stages of osteogenesis in vitro. For this, osteoblastic-like cells (Saos-2 cell lineage) were irradiated in two different periods: during the Proliferation phase (PP; from the second to the fourth day) and during the Differentiation phase (DP; from the seventh to the ninth day). The energy density used in the study was 1.5 J/ cm2. The following parameters were evaluated: 1) quantification of collagen type 1 (COL 1), osteopontin (OPN), and bone morphogenetic protein 2 (BMP-2); 2) quantification of alkaline phosphatase (ALP) activity; and 3) quantification of extracellular matrix (ECM) mineralization. Non-irradiated cultures were used as controls. The data were analyzed using the Student's t-test or one-way ANOVA, considering a significance level of 5%. The results indicated that COL 1 and BMP-2 quantification was higher in Saos-2 irradiated during the DP in relation to the control group at day 10 (p < 0.05). No differences were observed for other comparisons at this time point (p > 0.05). OPN expression was greater in PP compared with the other experimental groups at day 10 (p < 0.05). Irradiation did not affect ALP activity in Saos-2 regardless of the exposure phase and the time point evaluated (p > 0.05). At day 14, ECM mineralization was higher in Saos-2 cultures irradiated during the DP in relation to the PP (p < 0.05). In conclusion, the results suggested that the effects of PBM on osteoblastic cells may be influenced by the stage of cell differentiation.
Asunto(s)
Fosfatasa Alcalina , Proteína Morfogenética Ósea 2 , Diferenciación Celular , Proliferación Celular , Colágeno Tipo I , Terapia por Luz de Baja Intensidad , Osteoblastos , Osteogénesis , Osteopontina , Osteogénesis/efectos de la radiación , Humanos , Proteína Morfogenética Ósea 2/metabolismo , Fosfatasa Alcalina/metabolismo , Osteopontina/metabolismo , Diferenciación Celular/efectos de la radiación , Colágeno Tipo I/metabolismo , Osteoblastos/efectos de la radiación , Osteoblastos/citología , Osteoblastos/metabolismo , Proliferación Celular/efectos de la radiación , Matriz Extracelular/metabolismo , Matriz Extracelular/efectos de la radiaciónRESUMEN
Various biological effects of ionizing radiation, especially continuous exposure to low-dose radiation (LDR), have attracted considerable attention. Impaired bone structure caused by LDR has been reported, but little is known about the mechanism involved in the disruption of bone metabolism. In this study, given that LDR was found to (at a cumulative dose of 0.10â¯Gy) disturb the serum Mg2+ level and Notch1 signal in the mouse femur tissues, the effects of LDR on osteogenesis and the underlying molecular mechanisms were investigated based on an in vitro culture system for bone marrow stromal cells (BMSCs). Our data showed that cumulative LDR suppressed the osteogenic potential in BMSCs as a result of upregulation of Notch1 signaling. Further analyses indicated that the upregulation of NICD1 (Notch1 intracellular domain), the key intracellular domain for Notch1 signaling, under LDR was a consequence of enhanced protein stabilization caused by SUMOylation (small ubiquitin-like modification). Specifically, the downregulation of SENP1 (sentrin/SUMO-specific protease 1) expression induced by LDR enhanced the SUMOylation of NICD1, causing the accumulation of Notch1 signaling, which eventually inhibited the osteogenic potential of BMSCs. In conclusion, this work expounded on the mechanisms underlying the impacts of LDR on bone metabolism and shed light on the research on bone regeneration under radiation.
Asunto(s)
Diferenciación Celular , Células Madre Mesenquimatosas , Osteogénesis , Receptor Notch1 , Sumoilación , Animales , Osteogénesis/efectos de la radiación , Ratones , Sumoilación/efectos de la radiación , Receptor Notch1/metabolismo , Receptor Notch1/genética , Células Madre Mesenquimatosas/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Transducción de Señal/efectos de la radiación , Masculino , Fémur/efectos de la radiación , Relación Dosis-Respuesta en la RadiaciónRESUMEN
PURPOSE: Laser irradiation activates a range of cellular processes in the periodontal components and promotes tissue repair. However, its effect on osteogenic differentiation of human cementoblast lineage cells remains unclear. This study aimed to examine the effects of high-frequency semiconductor laser irradiation on the osteogenic differentiation of human cementoblast lineage (HCEM) cells. METHODS: HCEM cells were cultured to reach 80% confluence and irradiated with a gallium-aluminum-arsenide (Ga-Al-As) semiconductor laser with a pulse width of 200 ns and wavelength of 910 at a dose of 0-2.0 J/cm2. The outcomes were assessed by analyzing the mRNA levels of alkaline phosphatase (ALP), runt-related transcription factor 2 (RUNX2), and type I collagen (COLL1) using real-time polymerase chain reaction (PCR) analysis 24 h after laser irradiation. Cell mineralization was evaluated using ALP activity, calcium deposition, and Alizarin Red staining. RESULTS: The laser-irradiated HCEM cells showed significantly enhanced gene expression levels of ALP, RUNX2, and COLL1 as well as ALP activity and calcium concentration in the culture medium compared with the non-irradiated cells. In addition, enhanced calcification deposits were confirmed in the laser-irradiated group compared with the non-irradiated group at 21 and 28 days after the induction of osteogenic differentiation. CONCLUSION: High-frequency semiconductor laser irradiation enhances the osteogenic differentiation potential of cultured HCEM cells, underscoring its potential utility for periodontal tissue regeneration.
Asunto(s)
Diferenciación Celular , Cemento Dental , Láseres de Semiconductores , Osteogénesis , Humanos , Láseres de Semiconductores/uso terapéutico , Diferenciación Celular/efectos de la radiación , Osteogénesis/efectos de la radiación , Cemento Dental/efectos de la radiación , Cemento Dental/citología , Fosfatasa Alcalina/metabolismo , Células Cultivadas , Terapia por Luz de Baja Intensidad/métodos , Subunidad alfa 1 del Factor de Unión al Sitio Principal/metabolismo , Subunidad alfa 1 del Factor de Unión al Sitio Principal/genética , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismoRESUMEN
The Ti6Al4V (TC4) alloy, a prevalent biomedical material in orthopedics, still faces limitation of the insufficient osseointegration. To improve the bioactivity of TC4, introducing the electric environment onto the TC4 surface may be an effective way in the view of the necessity of endogenous electric microenvironment in bone regeneration. Herein, a Volta potential pattern was engendered on the TC4 surface via parallel laser patterning, so as to promote the osteogenic differentiation of cells. A 15 W laser successfully transformed the original α + ß dual phase towards radially distributed lath-like martensite phase in the laser treated region. The atomic lattice distortion between the heterogeneous microstructures of the laser treated and untreated regions leads to a significant Volta potential fluctuation on the TC4 surface. The Volta potential pattern as well as the laser-engraved microgrooves respectively induced mutually orthogonal cell alignments. The hBMSCs osteogenic differentiation was significantly enhanced on the laser treated TC4 surfaces in comparison to the surface without the laser treatment. Moreover, a drastic Volta potential gradient on the TC4 surface (treated with 15 W power and 400 µm interval) resulted in the most pronounced osteogenic differentiation tendency compared to other groups. Modulating the electric environment on the TC4 surface by manipulating the phase transformation may provide an effective way in evoking favorable cell response of bone regeneration, thereby improving the bioactivity of TC4 implant.
Asunto(s)
Aleaciones , Diferenciación Celular , Rayos Láser , Células Madre Mesenquimatosas , Osteogénesis , Propiedades de Superficie , Titanio , Osteogénesis/efectos de la radiación , Osteogénesis/fisiología , Aleaciones/química , Titanio/química , Humanos , Células Madre Mesenquimatosas/citología , Células CultivadasRESUMEN
Orthopedic surgeons face a significant challenge in treating critical-size femoral defects (CSFD) caused by osteoporosis (OP), trauma, infection, or bone tumor resections. In this study for the first time, the application of photobiomodulation (PBM) and bone marrow mesenchymal stem cell-conditioned medium (BM-MSC-CM) to improve the osteogenic characteristics of mineralized bone scaffold (MBS) in ovariectomy-induced osteoporotic (OVX) rats with a CSFD was tested. Five groups of OVX rats with CSFD were created: (1) Control (C); (2) MBS; (3) MBS + CM; (4) MBS + PBM; (5) MBS + CM + PBM. Computed tomography scans (CT scans), compression indentation tests, and histological and stereological analyses were carried out after euthanasia at 12 weeks following implantation surgery. The CT scan results showed that CSFD in the MBS + CM, MBS + PBM, and MBS + CM + PBM groups was significantly smaller compared to the control group (p = 0.01, p = 0.04, and p = 0.000, respectively). Moreover, the CSFD size was substantially smaller in the MBS + CM + PBM treatment group than in the MBS, MBS + CM, and MBS + PBM treatment groups (p = 0.004, p = 0.04, and p = 0.01, respectively). The MBS + PBM and MBS + CM + PBM treatments had significantly increased maximum force relative to the control group (p = 0.01 and p = 0.03, respectively). Bending stiffness significantly increased in MBS (p = 0.006), MBS + CM, MBS + PBM, and MBS + CM + PBM treatments (all p = 0.004) relative to the control group. All treatment groups had considerably higher new trabecular bone volume (NTBV) than the control group (all, p = 0.004). Combined therapies with MBS + PBM and MBS + CM + PBM substantially increased the NTBV relative to the MBS group (all, p = 0.004). The MBS + CM + PBM treatment had a markedly higher NTBV than the MBS + PBM (p = 0.006) and MBS + CM (p = 0.004) treatments. MBS + CM + PBM, MBS + PBM, and MBS + CM treatments significantly accelerated bone regeneration of CSFD in OVX rats. PBM + CM enhanced the osteogenesis of the MBS compared to other treatment groups.
Asunto(s)
Terapia por Luz de Baja Intensidad , Células Madre Mesenquimatosas , Animales , Ratas , Terapia por Luz de Baja Intensidad/métodos , Medios de Cultivo Condicionados , Femenino , Ratas Sprague-Dawley , Fémur/efectos de la radiación , Fémur/diagnóstico por imagen , Tomografía Computarizada por Rayos X , Osteoporosis/radioterapia , Osteoporosis/terapia , Ovariectomía , Andamios del Tejido , Osteogénesis/efectos de la radiación , Regeneración Ósea/efectos de la radiaciónRESUMEN
AIM: Radiotherapy is associated with cell depletion and loss of blood supply, which are linked to compromised bone healing. However, the molecular events underlying these effects at the tissue-implant interface have not been fully elucidated. This study aimed to determine the major molecular mediators associated with compromised osseointegration due to previous exposure to radiation. MATERIALS AND METHODS: Titanium implants were placed in rat tibiae with or without pre-exposure to 20 Gy irradiation. Histomorphometric, biomechanical, quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay analyses were performed at 1 and 4 weeks after implantation. RESULTS: The detrimental effects of irradiation were characterized by reduced bone-implant contact and removal torque. Furthermore, pre-exposure to radiation induced different molecular dysfunctions such as (i) increased expression of pro-inflammatory (Tnf) and osteoclastic (Ctsk) genes and decreased expression of the bone formation (Alpl) gene in implant-adherent cells; (ii) increased expression of bone formation (Alpl and Bglap) genes in peri-implant bone; and (iii) increased expression of pro-inflammatory (Tnf) and pro-fibrotic (Tgfb1) genes in peri-implant soft tissue. The serum levels of pro-inflammatory, bone formation and bone resorption proteins were greater in the irradiated rats. CONCLUSIONS: Irradiation causes the dysregulation of multiple biological activities, among which perturbed inflammation seems to play a common role in hindering osseointegration.
Asunto(s)
Oseointegración , Tibia , Animales , Oseointegración/efectos de la radiación , Ratas , Tibia/efectos de la radiación , Masculino , Implantes Dentales , Titanio , Interfase Hueso-Implante , Ratas Wistar , Implantación Dental Endoósea , Osteogénesis/efectos de la radiaciónRESUMEN
Background: This investigation set out to compare the impacts of low-level diode laser (LLDL) and red light-emitting diode (LED) on the survival of human dental pulp stem cells (hDPSCs) and osteogenic/odontogenic differentiation. Methods and materials: In this ex vivo experimental study, the experimental groups underwent the irradiation of LLDL (4 J/cm2 energy density) and red LED in the osteogenic medium. Survival of hDPSCs was assessed after 24 and 48 h (n = 9) using the methyl thiazolyl tetrazolium (MTT) assay. The assessment of osteogenic/odontogenic differentiation was conducted using alizarin red staining (ARS; three repetitions). The investigation of osteogenic and odontogenic gene expression was performed at two time points, specifically 24 and 48 h (n = 12). This analysis was performed utilizing real-time reverse-transcription polymerase chain reaction (RT-PCR). The groups were compared at each time point using SPSS version 24. To analyze the data, the Mann-Whitney U test, analysis of variance, Tukey's test, and t-test were utilized. Results: The MTT assay showed that LLDL significantly decreased the survival of hDPSCs after 48 h, compared with other groups (p < 0.05). The qualitative results of ARS revealed that LLDL and red LED increased the osteogenic differentiation of hDPSCs. LLDL and red LED both upregulated the expression of osteogenic/odontogenic genes, including bone sialoprotein (BSP), alkaline phosphatase (ALP), dentin matrix protein 1 (DMP1), and dentin sialophosphoprotein (DSPP), in hDPSCs. The LLDL group exhibited a higher level of gene upregulation (p < 0.0001). Conclusions: The cell survival of hDPSCs was reduced, despite an increase in osteogenic/odontogenic activity. Clinical relevance: Introduction of noninvasive methods in regenerative endodontic treatments.
Asunto(s)
Diferenciación Celular , Supervivencia Celular , Pulpa Dental , Láseres de Semiconductores , Terapia por Luz de Baja Intensidad , Odontogénesis , Osteogénesis , Células Madre , Humanos , Pulpa Dental/citología , Pulpa Dental/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Osteogénesis/efectos de la radiación , Células Madre/efectos de la radiación , Células Madre/citología , Supervivencia Celular/efectos de la radiación , Odontogénesis/efectos de la radiación , Células Cultivadas , Luz RojaRESUMEN
Pulsed electromagnetic field (PEMF) stimulation has been widely applied clinically to promote bone healing; however, its detailed mechanism of action, particularly in endochondral ossification, remains elusive, and long-term stimulation is required for its satisfactory effect. The aim of this study was to investigate the involvement of the mammalian target of rapamycin (mTOR) pathway in chondrocyte differentiation and proliferation using a mouse prechondroblast cell line (ATDC5), and establish an efficient PEMF stimulation strategy for endochondral ossification. The changes in cell differentiation (gene expression levels of aggrecan, type II collagen, and type X collagen) and proliferation (cellular uptake of bromodeoxyuridine [BrdU]) in ATDC5 cells in the presence or absence of rapamycin, an mTOR inhibitor, was measured. The effects of continuous and intermittent PEMF stimulation on changes in cell differentiation and proliferation were compared. Rapamycin significantly suppressed the induction of cell differentiation markers and the cell proliferation activity. Furthermore, only intermittent PEMF stimulation continuously activated the mTOR pathway in ATDC5 cells, significantly promoting cell proliferation. These results demonstrate the involvement of the mTOR pathway in chondrocyte differentiation and proliferation and suggest that intermittent PEMF stimulation could be effective as a stimulus for endochondral ossification during fracture healing process, thereby reducing stimulation time.
Asunto(s)
Diferenciación Celular , Proliferación Celular , Condrocitos , Campos Electromagnéticos , Osteogénesis , Serina-Treonina Quinasas TOR , Animales , Ratones , Osteogénesis/efectos de la radiación , Condrocitos/citología , Condrocitos/metabolismo , Condrocitos/fisiología , Línea Celular , Serina-Treonina Quinasas TOR/metabolismo , Sirolimus/farmacología , Cartílago/metabolismo , Cartílago/citología , Cartílago/fisiología , Transducción de Señal , Regulación de la Expresión Génica/efectos de la radiaciónRESUMEN
The effect of near infrared (NIR) laser irradiation on proliferation and osteogenic differentiation of buccal fat pad-derived stem cells and the role of transient receptor potential (TRP) channels was investigated in the current research. After stem cell isolation, a 940 nm laser with 0.1 W, 3 J/cm2 was used in pulsed and continuous mode for irradiation in 3 sessions once every 48 h. The cells were cultured in the following groups: non-osteogenic differentiation medium/primary medium (PM) and osteogenic medium (OM) groups with laser-irradiated (L +), without irradiation (L -), laser treated + Capsazepine inhibitor (L + Cap), and laser treated + Skf96365 inhibitor (L + Skf). Alizarin Red staining and RT-PCR were used to assess osteogenic differentiation and evaluate RUNX2, Osterix, and ALP gene expression levels. The pulsed setting showed the best viability results (P < 0.05) and was used for osteogenic differentiation evaluations. The results of Alizarin red staining were not statistically different between the four groups. Osterix and ALP expression increased in the (L +) group. This upregulation abrogated in the presence of Capsazepine, TRPV1 inhibitor (L + Cap); however, no significant effect was observed with Skf96365 (L + Skf).
Asunto(s)
Tejido Adiposo , Células Madre , Canales de Potencial de Receptor Transitorio , Humanos , Tejido Adiposo/efectos de la radiación , Diferenciación Celular/genética , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Células Cultivadas , Osteogénesis/genética , Osteogénesis/efectos de la radiación , Células Madre/efectos de la radiación , Canales de Potencial de Receptor Transitorio/metabolismo , Rayos InfrarrojosRESUMEN
Low-level laser therapy (LLLT) also known as photobiomodulation is a treatment to change cellular biological activity. The exact effects of LLLT remain unclear due to the different irradiation protocols. The purpose of this study was to investigate the effects of LLLT by three different irradiation methods on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro. BMSCs were inoculated in 24-well plates and then irradiated or not (control) with a laser using three different irradiation methods. The irradiation methods were spot irradiation, covering irradiation, and scanning irradiation according to different spot areas (0.07 cm2 or 1.96 cm2) and irradiation areas (0.35 cm2 or 1.96 cm2), respectively. The laser was applied three times at energy densities of 4 J/cm2. The cell proliferation by CCK-8. ALP activity assay, alizarin red, and quantitative real-time polymerase chain reaction (RT-PCR) were performed to assess osteogenic differentiation and mineralization. Increases in cell proliferation was obvious following irradiation, especially for covering irradiation. The ALP activity was significantly increased in irradiated groups compared with non-irradiated control. The level of mineralization was obviously improved following irradiation, particularly for covering irradiation. RT-PCR detected significantly higher expression of ALP, OPN, OCN, and RUNX-2 in the group covering than in the others, and control is the lowest. The presented results indicate that the biostimulative effects of LLLT on BMSCs was influenced by t he irradiation method, and the covering irradiation is more favorable method to promote the proliferation and osteogenic differentiation of BMSCs.
Asunto(s)
Terapia por Luz de Baja Intensidad , Células Madre Mesenquimatosas , Osteogénesis/genética , Osteogénesis/efectos de la radiación , Células de la Médula Ósea , Células Madre Mesenquimatosas/efectos de la radiación , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Células CultivadasRESUMEN
Human dental pulp stem cells (hDPSCs) have attracted tremendous attention in tissue regeneration engineering due to their excellent multidirectional differentiation potential. Photobiomodulation (PBM) using low-level light-emitting diodes (LEDs) or lasers has been proved to promote the osteogenesis of mesenchymal stem cells. However, the effect of LEDs on osteogenic differentiation of hDPSCs has little published data. In this work, the effect of blue LEDs with different energy densities of 2, 4, 6, 8, 10 J/cm2 on osteogenic differentiation of hDPSCs was examined by using in vitro ALP staining, ALP activity, mineralization, and real-time PCR. The results showed that compared with the control group, osteogenic differentiation was significantly enhanced in blue LEDs treated groups. As the energy density increased, the level of osteogenesis initially increased and then decreased reaching the highest level at 6 J/cm2. Transient receptor potential vanilloid 1 (TRPV1), a Ca2+ ion channel, was believed to be a potential player in osteogenesis by photobiomodulation. By immunofluorescence assay, calcium influx assay, PCR, and ALP staining, it was shown that blue LEDs irradiation can increase the activity of TRPV1 and intracellular calcium levels similarly to the agonist of TRPV1 capsaicin. Additionally, pretreatment with capsazepine, a selective TRPV1 inhibitor, was able to abrogate the osteogenic effect of blue LEDs. In conclusion, these findings proposed that blue LEDs can promote the osteogenesis of hDPSCs within the appropriate range (4-8 J/cm2) during culture of osteogenic medium, and TRPV1/Ca2+ may be an essential signaling pathway involved in blue LEDs-induced osteogenesis, providing new insights for the use of hDPSCs in tissue regeneration engineering.
Asunto(s)
Células Madre Mesenquimatosas , Osteogénesis , Calcio/metabolismo , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Células Cultivadas , Pulpa Dental , Humanos , Osteogénesis/efectos de la radiaciónRESUMEN
The biological aging of titanium implants affects the service lifetime negatively in clinical applications, and Ultraviolet (UV) irradiation is an applicable method to overcome the biological aging. This study investigated the changes in surface characteristics and biological properties of bioactive titanium surfaces with different structure and topography after Ultraviolet C (UVC) irradiation. The bioactive titanium surfaces were prepared by anodizing (AO), sandblasting and acid-etching (SLA), acid-alkali etching (AA), alkali-heat etching (AH) methods. Samples were stored at dark for 7 weeks to simulate biological aging process and then irradiated by UVC for 2 h. The results showed that the hydroxyl groups (Ti-OH) on surfaces, which are crucial to enhance the biological properties, were easier to be generated on AO surfaces by UVC-irradiation, owing to a mixture of anatase and rutile on surfaces. UVC-irradiation had the strongest effect on AO surfaces to enhance the bioactivity in bone-like apatite deposition and better biocompatibility in mesenchymal stem cells (MSCs) attachment and proliferation. Therefore, titanium surfaces with a mixture phase of anatase and rutile have the potential to effectively utilize the benefits of UVC-irradiation to overcome the negative effects of the biological aging and have a promising clinical application prospect.
Asunto(s)
Envejecimiento , Titanio , Rayos Ultravioleta , Envejecimiento/efectos de los fármacos , Envejecimiento/efectos de la radiación , Animales , Células Cultivadas , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Osteogénesis/efectos de los fármacos , Osteogénesis/efectos de la radiación , Conejos , Propiedades de SuperficieRESUMEN
BACKGROUND: Electromagnetic waves can cause biological effects on repair process. Due to the proximity of the jawbone and the soft tissue around it in a part of the face with which it has the closest contact during the cell phone use, this study aims to investigate the effect of mobile waves on socket healing after tooth extraction in rats. METHODS: This experimental study was conducted on 32 rats. The case group was exposed to a 900 MHz frequency electromagnetic field for 30 min/d. Sacrificed eight rats from the case group and 7 rats from the control group on day 14, and 8 rats from the case group and 7 rats from the control group were sacrificed on day 28 at the end of the radiation period and CBCT and microscopic examinations on the maxillary bones and soft tissue were performed. RESULTS: According to the findings, the healing process was significantly different in two groups in terms of the percentage of new bone formation on day 14 after the end of radiation (P=0.014). The other measured parameters including the degree of inflammation, thickness of the formed bone, number of osteoblasts and Gray Scale had no significant difference between the two groups in any of the 14-day and 28-day intervals. CONCLUSIONS: The results of this study showed that intermittent exposure to high frequency electromagnetic fields over a period of 20 hours has no significant effect on the healing process of alveolar socket after tooth extraction in rats.
Asunto(s)
Teléfono Celular , Alveolo Dental , Ratas , Animales , Extracción Dental , Ligamento Periodontal , Osteogénesis/efectos de la radiaciónRESUMEN
Photo-functionalization of titanium orthopedic/prosthetic implants using ultraviolet illumination is known to improve osteogenesis. Therefore, in this study, we aimed to examine the influence of vacuum ultraviolet (VUV)-treated titanium surfaces on osteoblast cell adhesion, activity, and differentiation. Osteoblastic cells were cultured on titanium substrates treated with various VUV treatment conditions (0, 6.2, 18.7, and 37.4 J/cm2) and their behavior was evaluated. The results revealed that cell adhesion was increased whereas cell activity and differentiation ability were decreased upon cell culture on VUV-treated substrates. In particular, cell activity and differentiation ability were dramatically suppressed with 18.7 J/cm2 VUV irradiation. Within the limitations of this cell-based experiment, we clarified the VUV treatment conditions in which cell adhesion was improved but cell activity and differentiation ability were suppressed. These results indicate that VUV-treatment can be used to influence cell growth properties and can be used to accelerate or suppress cell differentiation on implant substrates.
Asunto(s)
Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Osteogénesis/efectos de los fármacos , Titanio/farmacología , Adhesión Celular/efectos de los fármacos , Adhesión Celular/efectos de la radiación , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Humanos , Osteoblastos/efectos de los fármacos , Osteoblastos/efectos de la radiación , Osteogénesis/genética , Osteogénesis/efectos de la radiación , Especificidad por Sustrato , Propiedades de Superficie/efectos de los fármacos , Propiedades de Superficie/efectos de la radiación , Rayos Ultravioleta/efectos adversos , VacioRESUMEN
This study aims to evaluate the impact of red LED irradiation on the viability, proliferation, colonogenic potential, markers expression along with osteogenic and chondrogenic differentiation of dental pulp stem cells. DPSCs were isolated from sound human permanent teeth using enzymatic digestion method and seeded with regular culture media. Cells at P4 were irradiated using red LED Light (627 nm, 2 J/cm2) and examined for growth kinetics, and multilineage differentiation using the appropriate differentiation media. The irradiated groups showed an increase in cellular growth rates, cell viability, clonogenic potential, and decrease in population doubling time compared to the control group. Cells of the irradiated groups showed enhanced differentiation towards osteogenic and chondrogenic lineages as revealed by histochemical staining using alizarin red and alcian blue stains. Photobiomodulation is an emerging promising element of tissue engineering triad besides stem cells, scaffolds, and growth factors.
Asunto(s)
Terapia por Luz de Baja Intensidad , Diferenciación Celular/efectos de la radiación , Proliferación Celular/efectos de la radiación , Células Cultivadas , Pulpa Dental , Humanos , Cinética , Osteogénesis/efectos de la radiación , Células MadreRESUMEN
OBJECTIVE: To evaluate the effect of near infra-red gallium-aluminium-arsenide (GaAlAs) diode laser (805 nm) irradiation on proliferation and differentiation of rat femoral bone marrow-derived mesenchymal stem cells (BMSCs) cultured in osteogenic medium. MATERIALS AND METHODS: BMSCs were obtained from femurs of 60 Sprague Dawley rats (200 gm). The control group comprised isolated BMSCs supplemented with an osteogenic differentiation medium. On the other hand, in the experimental group, the BMSCs were irradiated with a near-infrared laser in addition to an osteogenic differentiation medium. The experimental group was irradiated with a soft tissue laser comprising of allium-aluminium-arsenic (Ga-Al-Ar) Diode at a near-infrared wavelength of 805 nm in continuous mode. The different output powers applied were 0.5 W, 1.0 W, 1.5 W and 2.0 W respectively. Various energy levels of 1, 4, 7 and 10 J were used for irradiation. Alkaline phosphatase (ALP) assay and Alizarin staining were performed to confirm osteogenic differentiation. Statistical analysis was done using a one-way ANOVA and a p-value of <0.05 was considered significant. RESULTS: According to our findings, 1.27 J/cm2 was the optimal energy density value that significantly increased the BMSC proliferation at the output of 1.5 W with the power density of 1.27 W/cm2. On 1.27 J/cm2, there was a significant difference compared to the control group on the first day, and the osteogenic differentiation increased significantly on the 4th day compared to the 1st day. CONCLUSIONS: According to our findings, 1.27 J/cm2 was the optimal energy density value that significantly increased the BMSC proliferation at the output of 1.5 W with the power density of 1.27 W/cm2.