Browsing by Author "Smith, Patricio C."

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    Ageing, dental caries and periodontal diseases
    (2017) López, R.; Smith, Patricio C.; Göstemeyer, G.; Schwendicke, F.
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    c-Jun N terminal kinase modulates NOX-4 derived ROS production and myofibroblasts differentiation in human breast stromal cells
    (2014) Arancibia, Rodrigo.; Smith, Patricio C.; Tobar, Nicolás.; Toyos, Marcela.; Urra, Carla.; Méndez, Nicolás.; Martínez, Jorge.
    Abstract Background Hard consistency, developed under the influence of tumor cell factors, is a characteristic feature of a breast tumor. Activation of resident fibroblasts leading to a myofibroblast phenotype is the principal feature that orchestrates this fibrotic process. The aim of this study was to assess the effects induced by TGF-β1, a growth factor abundantly present in tumor microenvironment, on the molecular mechanisms that mediate myofibroblastic differentiation of normal human mammary fibroblasts. Methods We used an immortalized fibroblastic cell line derived from normal mammary tissue (RMF-EG cells) to study the effect of TGF-β1 in the expression of α-SMA and CTGF as markers of myofibroblastic differentiation. The influence of redox status and JNK activity on TGF-β1-induced transcriptional activity was measured by a luciferase reporter assay. We also used a shRNA approach to evaluate the influence of NOX4 in myofibroblastic differentiation. Results TGF-β1 stimulates the expression of myofibroblast markers α-SMA and CTGF. Using a NOX inhibitor (DPI) and cells expressing a shRNA for NOX4, we demonstrated that TGF-β1 promotes an oxidative environment that favors myofibroblastic differentiation. We also found that activation of c-Jun N-terminal kinase is required for TGF-β1-dependent expression of CTGF, NOX4 and α-SMA. Conclusions Human mammary stromal fibrosis, evaluated by the expression of early and late markers as CTGF and α-SMA, depends on the activation of JNK signaling pathway. Our results show that JNK activation is an early event that precedes the increase in ROS levels leading to myofibroblastic differentiation and tumor fibrosis, suggesting that inhibition of JNK may be used a method to interrupt the development of tumor desmoplasia.
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    Cetylpyridinium chloride blocks herpes simplex virus replication in gingival fibroblasts
    (2020) Alvarez, Diana M.; Duarte, Luisa F.; Corrales, Nicolas; Smith, Patricio C.; Gonzalez, Pablo A.
    Infections with herpes simplex viruses are lifelong and highly prevalent worldwide. Individuals with clinical symptoms elicited by HSVs may suffer from occasional or recurrent herpetic lesions in the orofacial and genital areas. Despite the existence of nucleoside analogues that interfere with HSV replication, such as acyclovir, these drugs are somewhat ineffective in treating skin lesions as topical formulations only reduce in one or few days the duration of the herpetic ulcers. Cetylpyridinium chloride (CPC) is a quaternary ammonium compound present in numerous hygiene products, such as mouthwashes, deodorants, aphtae-treating formulations and oral tablets as an anti-septic to limit bacterial growth. Some reports indicate that CPC can also modulate host signaling pathways, namely NF-kappa B signaling. Because HSV infection is modulated by NF-kappa B, we sought to assess whether CPC has antiviral effects against HSVs. Using wild-type HSV-1 and HSV-2, as well as viruses that are acyclovirresistant or encode GFP reporter genes, we assessed the antiviral capacity of CPC in epithelial cells and human gingival fibroblasts expanded from the oral cavity and its mechanism of action. We found that a short, 10-min exposure to CPC added after HSV entry into the cells, significantly limited viral replication in both cell types by impairing viral gene expression. Interestingly, our results suggest that CPC blocks HSV replication by interfering with the translocation of NF-kappa B into the nucleus of HSV-infected cells. Taken together, these findings suggest that formulations containing CPC may help limit HSV replication in infected tissues and consequently reduce viral shedding.
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    Chitosan and platelet-derived growth factor synergistically stimulate cell proliferation in gingival fibroblasts
    (2013) Silva, Daniel; Arancibia, Rodrigo; Tapia, Cristián; Acuña Rougier, Cristina; Díaz Dosque, Mario; Cáceres, Mónica; Martínez, J.; Smith, Patricio C.
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    Comparative effect of platelet-rich plasma, platelet-poor plasma, and fetal bovine serum on the proliferative response of periodontal ligament cell subpopulations
    (2019) Martinez, Constanza E.; Gomez, Roberto; Kalergis, Alexis M.; Smith, Patricio C.
    ObjectivesCell-based therapies involve the need to expand cell cultures ex vivo for their subsequent implantation in an autologous manner. An important limitation regarding this technology is the use of fetal bovine serum (FBS) that has critical safety limitations. Platelet-derived fractions represent an autologous source of growth factors that may be used for the expansion of these cell cultures. Periodontal ligament (PDL) cells comprise a heterogeneous cell population that may not necessarily respond in a uniform manner to proliferative stimuli. The aim of this study was to evaluate the ability of two platelet-derived fractions, platelet-rich plasma (PRP) and platelet-poor plasma (PPP) and FBS on the proliferative response of different subpopulations of PDL cell cultures.Materials and methodsPDL cells were characterized and then exposed to PRP, PPP, or FBS during 2, 5, or 14days to analyze cell proliferation and clonogenic capability. Cell proliferation was evaluated through immunofluorescence for Ki67 and by tracing carboxyfluorescein diacetate succinimidyl ester (CFSE) dye in combination with mesenchymal stem cell markers using flow cytometry.ResultsBoth PRP and PPP stimulated PDL cell proliferation and their clonogenic ability. We found a significant increase of CD73- and CD90-positive cells after PRP or PPP treatment, compared to FBS. Otherwise, no differences were found regarding the response of CD146-or CD105-positive cells when stimulated with PRP, PPP, or FBS.ConclusionPRP and PPP can stimulate the proliferation and clonogenicity of PDL cell populations including cells positive for CD90 and CD73 markers.Clinical relevanceThese findings may have implications for future therapies aiming to stimulate periodontal regeneration using autologous growth factors.
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    Cytokine profiles and the dynamic of gingivitis development in humans
    (2022) Leite, Fabio R. M.; Nascimento, Gustavo G.; Moller, Holger J.; Belibasakis, Georgios N.; Bostanci, Nagihan; Smith, Patricio C.; Lopez, Rodrigo
    Aim To investigate the relationship between cytokine profiles and "fast" and "slow" patterns of gingival inflammation development. Materials and Methods Forty-two adults participated in an experimental gingivitis study, comprising a 2-week hygiene phase (clinical examination and professional cleaning); a 3-week induction phase (absence of oral hygiene); and a 2-week resolution phase (re-establishment of oral hygiene). Plaque and gingival inflammation scores were assessed. Interferon-gamma (IFN-gamma), interleukin (IL)-1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12p70, IL-13, and tumour necrosis factor-alpha (TNF-alpha) from gingival crevicular fluid were collected and measured by multiplex ELISA. Group-based-trajectory-modelling (GBTM) was used to model cytokine profiles over the induction phase. The effect of gingival inflammation on cytokine levels over time was estimated with mixed-effects modelling. Results GBTM analysis revealed two cytokine profiles, "non-organized response" (IL-4, IL-6, IL-8, IL-12, and IL-13) and "organized response" (IL-2, IL-10, and TNF-alpha). Among the "slow" responders, neither cytokine profile was associated with gingivitis. In contrast, a "fast" response was associated with a higher "non-organized response" factor (coef. 0.14) and a lower "organized response" factor (coef. -0.03). Conclusion A "fast" gingivitis development was associated with a higher "non-organized response" and a lower "organized response", which may elucidate the role of individual variability in gingivitis susceptibility.
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    Defective Wound-healing in Aging Gingival Tissue
    (2014) Cáceres, M.; Oyarzún, A.; Smith, Patricio C.
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    Effects of Chitosan Particles in Periodontal Pathogens and Gingival Fibroblasts
    (2013) Arancibia Reyes, Rodrigo Esteban; Maturana Salgado, Cristián Roberto; Silva Vargas, Daniel Ignacio; Tobar Chler, Nicólas Andrés; Tapia Villanueva, Cristián Adolfo; Salazar, Juan C.; Martínez Winkler, Jorge; Smith, Patricio C.
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    FOXO1 regulates wound-healing responses in human gingival fibroblasts
    (2024) Rojas, Leticia; Tobar, Nicolas; Espinoza, Javier; Rios, Susana; Martinez, Constanza; Martinez, Jorge; Graves, Dana T.; Smith, Patricio C.
    Background and objective: Forkhead box-O 1 (FOXO1) is a transcription factor actively involved in oral wound healing at the epithelial barrier. However, less is known regarding the role of FOXO1 during the tissue repair response in the connective tissue compartment. This study explored the involvement of FOXO1 in the modulation of fibroblast activity related to wound healing. Methods: Primary cultures of human gingival fibroblasts were obtained from four healthy young donors. Myofibroblastic differentiation, collagen gel contraction, cell migration, cell spreading, and integrin activation were evaluated in the presence or absence of a FOXO1 inhibitor (AS1842856). Variations in mRNA and proteins of interest were evaluated through qRT-PCR and western blot, respectively. Distribution of actin, alpha-smooth muscle actin, and beta 1 integrin was evaluated using immunofluorescence. FOXO1 and TGF-beta 1 expression in gingival wound healing was assessed by immunohistochemistry in gingival wounds performed in C57BL/6 mice. Images were analyzed using ImageJ/Fiji. ANOVA or Kruskal-Wallis test followed by Tukey's or Dunn's post-hoc test was performed. All data are expressed as mean +/- SD. p < .05 was considered statistically significant. Results: FOXO1 inhibition caused a decrease in the expression of the myofibroblastic marker alpha-SMA along with a reduction in fibronectin, type I collagen, TGF-beta 1, and beta 1 integrin mRNA level. The FOXO1 inhibitor also caused decreases in cell migration, cell spreading, collagen gel contraction, and beta 1 integrin activation. FOXO1 and TGF-beta 1 were prominently expressed in gingival wounds in fibroblastic cells located at the wound bed. Conclusion: The present study indicates that FOXO1 plays an important role in the modulation of several wound-healing functions in gingival fibroblast. Moreover, our findings reveal an important regulatory role for FOXO1 on the differentiation of gingival myofibroblasts, the regulation of cell migration, and collagen contraction, all these functions being critical during tissue repair and fibrosis.
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    Galectin-8 induces partial epithelial–mesenchymal transition with invasive tumorigenic capabilities involving a FAK/EGFR/proteasome pathway in Madin–Darby canine kidney cells
    (2018) Oyanadel, Claudia; Holmes Videla, Christopher Edward; Pardo Huguet, Evelyn Cristina; Retamal Villarroel, Claudio Enrique; Shaughnessy, Ronan Patrick; Smith, Patricio C.; Cortés Martínez, Priscilla Rocío; Bravo Zehnder, Marcela; Metz Baer, Claudia Andrea; Feuerhake, Teo; Romero, Diego; Roa Strauch, Juan Carlos Enrique; Montecinos, Viviana; Soza Gajardo, Andrea; González, Alfonso
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    Glucose Promotes a Pro‐Oxidant and Pro‐Inflammatory Stromal Microenvironment Which Favors Motile Properties in Breast Tumor Cells
    (2017) Kallens, Violeta; Tobar, Nicolás; Molina, Jessica; Bidegain, Arantzazú; Smith, Patricio C.; Porras, Omar; Martínez, Jorge
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    Glycated Collagen Stimulates Differentiation of Gingival Myofibroblasts.
    (2017) Retamal, Ignacio N.; Hernández, Romina; Melo Ledermann, Francisco Javier; Zapata, Paulina; Martinez, Constanza; Martínez, Jorge; Smith, Patricio C.
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    Modulation of Mammary Stromal Cell Lactate Dynamics by Ambient Glucose and Epithelial Factors
    (2017) Tobar, N; Porras, O.; Smith, Patricio C.; Barros, L.; Martinez, J.
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    Research on growth factors in periodontology
    (2015) Smith, Patricio C.; Martínez, Constanza; Cáceres, Mónica; Martínez Castillo, Jorge
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    Smoking habits do not affect biological responses induced by leucocyte and platelet-rich fibrin in periodontal ligament cells
    (2023) Rios, Susana; Alvarez, Simon; Smith, Patricio C.; Saez, Claudia G.; Andrade, Catherine; Pinto, Nelson; Martinez, Constanza E.
    Background and Objective: Leucocyte- and platelet-rich fibrin has been developed to stimulate wound healing response. However, it is currently unknown whether smoking affects the biological responses elicited by leucocyte- and platelet-rich fibrin on periodontal ligament-derived mesenchymal stromal cells. This study analyzes the kinetics of biomolecule release from leucocyte- and platelet-rich fibrin derived from smokers and nonsmokers and their effect on periodontal ligament cell proliferation and migration as essential biological activities during wound healing. Methods: Biomolecules present in leucocyte- and platelet-rich fibrin exudates and conditioned media collected from smokers and nonsmokers were analyzed by Luminex arrays. Periodontal ligament-derived mesenchymal stromal cell obtained from one nonsmoker were treated with leucocyte- and platelet-rich fibrin exudates or leucocyte- and platelet-rich fibrin conditioned media derived from both smokers and nonsmokers. The parameters evaluated included cell proliferation, determined by Ki67 immunostaining and migration assessed using transwell assays. Also, cells were treated with nicotine in the presence of fetal bovine serum 10% or leucocyte- and platelet-rich fibrin conditioned media. Results: A similar biomolecular profile was detected in leucocyte- and platelet-rich fibrin exudates and leucocyte- and platelet-rich fibrin conditioned media from smokers and nonsmokers, stimulating (periodontal ligament-derived mesenchymal stromal cell) proliferation, and migration to a comparable degree. Nicotine reduced cell proliferation and migration of periodontal cells; however, this effect was recovered in the presence of leucocyte- and platelet-rich fibrin conditioned media. Conclusion: Leucocyte- and platelet-rich fibrin derived from smokers could be an autologous source of biomolecules to stimulate cell biological activities involved in wound healing in smokers who have difficulties in ceasing this habit. Clinical trials are required to evaluate the impact of leucocyte- and platelet-rich fibrin on healing responses in smokers.
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    Soluble MMP-14 produced by bone marrow-derived stromal cells sheds epithelial endoglin modulating the migratory properties of human breast cancer cells
    (2014) Tobar, N.; Avalos M., Celeste; Mendez, N; Smith, Patricio C.; Bernabeu, C.; Quintanilla, M.; Martínez, J.
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