Human Oral Fibroblasts Research Articles

Comparative evaluation of human oral fibroblast proliferation on 3D-printed zirconia and silicon nitride as new ceramic materials for implant abutment.

Cell adhesion and subsequent proliferation on material surfaces depend on the physical and chemical characteristics of the material. There is a lack of literature on human gingival fibroblast proliferation on comparatively newer additively manufactured materials like silicon nitride. This study focused on the physical characteristics of the materials with the aim to compare the adhesion and proliferation of human gingival fibroblasts on additively manufactured silicon nitride (AMSN) with additively manufactured zirconia, conventional milled titanium (MTi), and milled zirconia. Surface roughness and water contact angle were measured by profilometer and goniometer, respectively. CCK-8 assay was done to assess the cell growth at 24h (day 1), 48h (day 2), and 72h (day 3) in the same well. The morphologies of fibroblasts after cell attachment and proliferation were evaluated using scanning electron microscopy (SEM) after 72h. At the end of 24h (day 1) additively manufactured zirconia showed the best proliferation among the experimental groups, which was around 50% of the positive control group proliferation. There was no statistically significant difference among the experimental groups. At 48h (day 2) and 72h (day 3), a loss of cell growth was seen in almost all the experimental group wells. A positive cell proliferation on the AMSN was observed on day 3. Comparable cell proliferation was observed in the experimental groups. No conclusive correlation could be drawn between cell proliferation and surface roughness and water contact angle values in the experimental groups.

Arecoline promotes fibroblast activation and M2-macrophage polarization by up-regulating the expression of IL-4

ObjectiveTo determine the biological effects of arecoline on oral submucous fibrosis (OSF). DesignThe differential genes between OSF tissue and normal oral tissue were collected form GSE64216 dataset, analyzed by Gene Expression Omnibus (GEO) database. Real-time PCR and immunohistochemistry were used to analyze the expression of IL-4 gene and protein in oral tissue. Enzyme-linked immunosorbent assay (ELISA) was used to analyze the expression of exocrine IL-4 protein in human oral fibroblasts (HOF) pre-treated by arecoline. Cell Counting Kit-8 (CCK-8) and transwell assays were used to analyze the proliferation and migration of HOF cells, respectively. After IL-4 was knocked down by short hairpin (sh) plasmid, the proliferation and migration of HOF cells were detected. Flow cytometry was used to analyze the proportion of M2-macrophages. Real-time PCR and immunohistochemistry were used to verify the expression of biomarker proteins of macrophages in OSF tissues. ResultsThe expression of IL-4 gene and protein were both up-regulated in OSF tissue. Arecoline could enhance the expression of IL-4 gene and exocrine protein in HOF cells, and promote the proliferation and migration of HOF cells. While knockdown of IL-4 could inhibit arecoline-induced proliferation and migration in HOF cells. The results of flow cytometry showed that recombinant human IL-4 (rhIL-4) protein could increase the proportion of M2-macrophages. Similarly, the results of real-time PCR and immunohistochemistry showed the expression of ARG1 (Biomarker proteins of M2-macrophage) was up-regulated in OSF tissues. ConclusionArecoline promotes activation of fibroblasts and polarization of M2-macrophages by up-regulating the expression of IL-4.

The association between the expression level of nuclear paraspeckle assembly transcript 1 and the survival rate of head and neck cancer patients after treatment

Background/purposeThe long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) exhibits diverse and complicated functions in cancer progression. Despite reports suggesting both tumor-suppressive and oncogenic effects in various cancers, its specific role in head and neck squamous cell carcinoma (HNSCC) remains unclear. This study aimed to investigate the association between NEAT1 expression levels and survival outcomes in HNSCC patients. Materials and methodsPaired tissue samples of tumor and non-cancerous matching tissues (NCMT) from 92 HNSCC patients were collected. NEAT1 expression was analyzed using RT-qPCR. Clinical characteristics, treatment received, and survival rates of the patients were assessed to determine the correlation with NEAT1 expression and explore its association with alcohol, betel quid, and cigarette use. Additionally, we examined the effect of arecoline on NEAT1 expression in normal human oral keratinocytes (NHOK) and fibroblasts (NHOF). ResultsThe study revealed a significant downregulation of NEAT1 expression in oral cancer tissues compared to NCMT. Meanwhile, arecoline increased NEAT1 expression in NHOK and NHOF cells. However, patients with downregulated NEAT1 expression exhibited higher overall survival rates, particularly in those who did not receive chemotherapy or radiotherapy. ConclusionNEAT1 expression levels are associated with survival outcomes in HNSCC patients, with upregulated expression indicating a worse prognosis, suggesting this lncRNA might contribute to cancer aggressiveness, especially in the absence of active treatment. These findings indicate NEAT1 may serve as a potential prognostic biomarker in HNSCC, but further research is required to elucidate its role in cancer progression and its potential as a therapeutic target.

Gelselegine-gelsedine type bisindoles as well as the units from Gelsemium elegans with promoting proliferation of oral mucosa fibroblast cells

Two undescribed bisindole alkaloids, gelseginedine A (1) and its rearranged gelseginedine B (2), and seven unreported gelselegine-type oxindole alkaloids (3–9) were isolated from the stems and leaves of Gelsemium elegans, together with five known alkaloids (10–14). Compounds 1 and 2 represented the first examples of gelselegine-gelsedine type alkaloids which bridged two units by a double bond. Their structures with absolute configurations were elucidated by means of HRESIMS, NMR and calculational chemistry. The performed bioassay revealed that 14 could promote the proliferation of human oral mucosa fibroblast cells.

Antifibrotic Effect of Baicalin on Arecoline Induced Human Oral Fibroblast: An In-Vitro Study.

Baicalin is a flavonoid obtained from the Chinese herb Scutellaria baicalensis, which has a wide varieties of health benefits and scope to be studied for its therapeutic potential in oral fibrosis. The aim of the study was to investigate the antifibrotic effect of a Baicalin in arecoline induced human oral fibroblast in vitro setting. Arecoline and ethanolic extracts of Baicalin were commercially purchased from Sigma-Aldrich. Human oral fibroblasts were cultured and characterized with specific fibroblast markers, and cells were stimulated with arecoline. An MTT assay (3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide) was executed to determine the half-maximal inhibitory concentration of arecoline and Baicalin. Arecoline-induced cells (25µg/ml) were treated with a non-toxic dose of Baicalin (proliferative dose of 25µg/ml). Cytokine (CCL2, CXCL-8, IL17, IL-beta, and IL-6) and fibrotic marker genes were studied by reverse transcription-polymerase chain reaction (RT-PCR). The inhibitory effect of Baicalin was studied to prove its antifibrotic properties. Arecoline significantly upregulated all inflammatory and fibrotic markers. On treatment with 25µg/ml of Baicalin, all inflammatory and fibrotic markers were inhibited. Arecoline affects fibroblast morphology, supporting the fact that arecoline is cytotoxic to cells. Baicalin can be used as an antifibrotic herb to treat OSMF.

Adiponectin inhibits ROS/NLRP3 inflammatory pathway through FOXO3A to ameliorate oral submucosal fibrosis.

Oral submucous fibrosis (OSF) is an oral condition characterized by chronic progression, which may lead to the development of malignancy. Currently, available treatments for OSF only provide temporary relief of symptoms, and there is a limited availability of effective interventions that can effectively cure this condition. In this study, we aimed to investigate whether adiponectin (APN) could ameliorate OSF and the mechanisms involved in it. First, human oral mucosal fibroblasts (HOMFs) were cultured, an OSF model was established using arecoline, and APN and Imiquimod treatment were administered. Then we overexpressed NLRP3 and knocked down FOXO3A. FOXO3A, fibrosis-related factors (ɑ-SMA, COL1A, CTGF), TGF-β1/Smad3 signaling-related factors (TGF-β1, p-Smad3, Smad3), NLRP3 inflammasome-related factors (NLRP3, Caspase-1, IL-1β), and ROS levels were evaluated. Finally, we explored the effect of APN on OSF in mice by in vivo experiments. We found that arecoline significantly increased ɑ-SMA, COL1A, CTGF, and TGF-β1 expressions and promoted Smad3 phosphorylation, while APN significantly inhibited the elevation of these fibrosis-related factors. ROS production was significantly elevated in HOMFs after arecoline treatment, while APN treatment inhibited ROS production. However, the addition of Imiquimod and overexpression of NLRP3 exhibited a trend of elevated ROS, resisting the inhibitory effect of APN. Furthermore, adding Imiquimod and overexpression of NLRP3 elevated ɑ-SMA, COL1A and CTGF and activated TGF-β1/Smad3 signaling pathway. Additionally, knockdown of FOXO3A enhanced APN-inhibited ɑ-SMA and COL1A. In vivo experiments further confirmed that APN ameliorated OSF in mice by inhibiting ROS/NLRP3 inflammatory pathway. In conclusion, APN ameliorated arecoline-induced OSF by promoting FOXO3A expression and downregulating the ROS/NLRP3 pathway.

Periplaneta americana extract promotes hard palate mucosal wound healing via the PI3K/AKT signaling pathway in male mice

ObjectivesThis study aimed to investigate the effect of Periplaneta americana extract, a traditional Chinese medicine, on hard palate mucosal wound healing and explore the underlying mechanisms. DesignHard palate mucosal wound model was established and the effects of Periplaneta americana extract on hard palate mucosal wound healing were investigated by stereomicroscopy observation and histological evaluation in vivo. Human oral keratinocytes and human gingival fibroblasts, which play key roles in hard palate mucosal wound healing, were selected as the main research cells in vitro. The effects of Periplaneta americana extract on cell proliferation, migration, and collagen formation were determined by cell counting kit-8 (CCK-8) assay, Transwell assay, and Van Gieson staining. The underlying mechanism was revealed by RNA sequencing, and results were verified by western blot assay. ResultsStereomicroscopy observation and H&E staining confirmed that Periplaneta americana extract accelerated the healing rate of hard palate mucosal wound (p < 0.001) in vivo. Transwell assay and Van Gieson staining assay showed that Periplaneta americana extract promoted the migration and collagen formation of human oral keratinocytes (p < 0.001) and human gingival fibroblasts (p < 0.001) in vitro. Mechanistically, RNA sequencing and western blot assay demonstrated that Periplaneta americana extract promoted hard palate mucosal wound healing via PI3K/AKT signaling, and the beneficial effects of Periplaneta americana extract were abrogated by the PI3K inhibitor LY294002. ConclusionsPeriplaneta americana extract shows promising effects for the promotion of hard palate mucosal wound healing and may be a novel candidate for clinical translation.

Vacuum Ultraviolet (VUV) Light Photofunctionalization to Induce Human Oral Fibroblast Transmigration on Zirconia.

Soft tissue adhesion and sealing around dental and maxillofacial implants, related prosthetic components, and crowns are a clinical imperative to prevent adverse outcomes of periodontitis and periimplantitis. Zirconia is often used to fabricate implant components and crowns. Here, we hypothesized that UV treatment of zirconia would induce unique behaviors in fibroblasts that favor the establishment of a soft tissue seal. Human oral fibroblasts were cultured on zirconia specimens to confluency before placing a second zirconia specimen (either untreated or treated with one minute of 172 nm vacuum UV (VUV) light) next to the first specimen separated by a gap of 150 µm. After seven days of culture, fibroblasts only transmigrated onto VUV-treated zirconia, forming a 2.36 mm volume zone and 5.30 mm leading edge. Cells migrating on VUV-treated zirconia were enlarged, with robust formation of multidirectional cytoplastic projections, even on day seven. Fibroblasts were also cultured on horizontally placed and 45° and 60° tilted zirconia specimens, with the latter configurations compromising initial attachment and proliferation. However, VUV treatment of zirconia mitigated the negative impact of tilting, with higher tilt angles increasing the difference in cellular behavior between control and VUV-treated specimens. Fibroblast size, perimeter, and diameter on day seven were greater than on day one exclusively on VUV-treated zirconia. VUV treatment reduced surface elemental carbon and induced superhydrophilicity, confirming the removal of the hydrocarbon pellicle. Similar effects of VUV treatment were observed on glazed zirconia specimens with silica surfaces. One-minute VUV photofunctionalization of zirconia and silica therefore promotes human oral fibroblast attachment and proliferation, especially under challenging culture conditions, and induces specimen-to-specimen transmigration and sustainable photofunctionalization for at least seven days.

Isolation, characterization, and fibroblast uptake of bacterial extracellular vesicles from Porphyromonas gingivalis strains.

Periodontitis is an inflammatory condition caused by bacteria and represents a serious health problem worldwide as the inflammation damages the supporting tissues of the teeth and may predispose to systemic diseases. Porphyromonas gingivalis is considered a keystone periodontal pathogen that releases bacterial extracellular vesicles (bEVs) containing virulence factors, such as gingipains, that may contribute to the pathogenesis of periodontitis. This study aimed to isolate and characterize bEVs from three strains of P. gingivalis, investigate putative bEV uptake into human oral fibroblasts, and determine the gingipain activity of the bEVs. bEVs from three bacterial strains, ATCC 33277, A7A1-28, and W83, were isolated through ultrafiltration and size-exclusion chromatography. Vesicle size distribution was measured by nano-tracking analysis (NTA). Transmission electron microscopy was used for bEV visualization. Flow cytometry was used to detect bEVs and gingipain activity was measured with an enzyme assay using a substrate specific for arg-gingipain. The uptake of bEVs into oral fibroblasts was visualized using confocal microscopy. NTA showed bEV concentrations from 108 to 1011 particles/mL and bEV diameters from 42 to 356 nm. TEM pictures demonstrated vesicle-like structures. bEV-gingipains were detected both by flow cytometry and enzyme assay. Fibroblasts incubated with bEVs labeled with fluorescent dye displayed intracellular localization consistent with bEV internalization. In conclusion, bEVs from P. gingivalis were successfully isolated and characterized, and their uptake into human oral fibroblasts was documented. The bEVs displayed active gingipains demonstrating their origin from P. gingivalis and the potential role of bEVs in periodontitis.

MiR-3529-3p from PDGF-BB-induced cancer-associated fibroblast-derived exosomes promotes the malignancy of oral squamous cell carcinoma

AimsThis study aims to explore the role of exosomes from cancer-associated fibroblasts (CAFs) induced by PDGF-BB in promoting the malignancy of oral squamous cell carcinoma (OSCC) and provide new insight into the mechanism of OSCC progression and its treatment.Main methodsExosomes were extracted from human oral mucosa fibroblasts (hOMFs) and CAFs. Differentially expressed miRNAs of exosomes between hOMFs and CAFs were analysed using high-throughput sequencing and self-programmed R software. Cal-27, a human tongue squamous carcinoma cell line, was treated with exosomes. Differentially expressed miRNAs between clinical cancer tissues and adjacent tissues and between hOMF and CAF exosomes were verified by qRT‒PCR. The effect of miR-3529-3p on Cal-27 cells was clarified by overexpressing or knocking down miR-3529-3p in Cal-27 cells. Sample expression and differentially expressed miRNA expression were compared between cancer and paracarcinoma tissues.Key findingsWe found that exosomes from CAFs (CAF-Exos) were internalized by tongue squamous carcinoma cells and promoted their proliferation, migration, invasion, and antiapoptotic effects. MiR-3529-3p was a significant differentially expressed miRNA between CAF-Exos and exosomes from hOMFs (hOMF-Exos). The overexpression of miR-3529-3p promoted proliferation, migration, and invasion and inhibited apoptosis of Cal-27 cells.SignificanceThis study explores the role of PDGF-BB-induced CAFs in promoting malignancy in OSCC. This study will provide new insight into the mechanism of OSCC progression and its treatment.

Electronic Cigarettes, Heated Tobacco Products, and Oral Health: A Systematic Review and Meta-Analysis

Smokers employing electronic nicotine delivery systems (ENDS) and heated tobacco products (HTP) are currently the most common types of smoking patients seen in the dental practice. Both types of smoking are currently viewed as less harmful than cigarette smoking. However, many studies already indicate that they could harm oral health. This systematic review and meta-analysis aimed to collect a comprehensive overview of the actual knowledge regarding ENDS and HTP from a clinical and a laboratory perspective. Publications available through PubMed, Embase, the Web of Science, Scopus, and Google Scholar were used to summarize the effects of ENDS and HTP on oral health. Six surveys on self-perceived gum disease (T2 = 9.47 I2 = 99.32%), three cross-sectional studies reporting the BOP score (T2 = 8.68 I2 = 99.13%), and four in vitro studies on apoptosis after vaping exposure in human oral fibroblasts (T 2 = 8.10 I2 = 91.50%) were separately analyzed. The risk of bias ranged from critical to low. Both ENDS and HTP seem to have detrimental effects on periodontal and peri-implant parameters, and laboratory tests confirmed the presence of carcinogenic and inflammatory biomarkers. flavored e-liquids may also be a caries risk factor. Comprehensive smoking counseling should be carried out with all types of smoking patients, investigating the type of habit in terms of duration, nicotine percentage, and additional flavorings. Additional research is necessary to assess the long-term effects of alternative tobacco products on oral health.

Green-Synthesized Silver Nanoparticles: Antifungal and Cytotoxic Potential for Further Dental Applications.

Fungal infections caused by Candida albicans (C. albicans) are one of the most prevalent types of oral disorders in the elderly. It has been reported that drug resistance to fungal pathogens poses a severe risk to global healthcare systems and public health. Therefore, the goal of this work is to investigate the cytotoxic and antifungal properties of silver nanoparticles (AgNPs) produced using three different natural extracts: Berzelia lanuginose, Helichrysum cymosum, and Searsia crenata. According to the UV-Vis results, the synthesized AgNPs via B. lanuginose, H. cymosum, and S. crenata show surface plasmonic resonance (SPR) peaks at 430, 440, and 428 nm, respectively. HR-TEM revealed different shapes for the nanoparticles within the size ranges of 16-20, 31-60, and 57-72 nm for B. lanuginose, H. cymosum, and S. crenata, respectively. Using a human oral fibroblast cell line, the cytotoxicity of both AgNPs and plant extracts was tested at concentrations of 0.007, 0.012, 0.025, and 0.062 mg/mL (buccal mucosa fibroblasts). The antifungal activity showed growth inhibition zones of approximately 18 mm, 18.67 mm, and 18.33 mm for the AgNPs conjugated with B. lanuginose, H. cymosum, and S. crenata, respectively. For the studied samples, the minimum inhibitory concentration (MIC50) was less than 0.015 mg/mL. The AgNPs exhibited antifungal activity that was concentration- and size-dependent. The results of this study offer new insights into the cytotoxicity and antifungal activity of the green-synthesized AgNPs.

Fibroblasts are the most suitable cell source for regenerative medicine due to their high intracellular fibroblast growth factor 2 content

In our previous study, we found that dry-preserved multilayered fibroblast cell sheets promoted angiogenesis and wound healing in a mouse ulcer model by releasing high levels of intracellular fibroblast growth factor 2 (FGF2), hepatocyte growth factor (HGF), and vascular endothelial growth factor (VEGF), from dried cells. In the present study, to identify which cell types are suitable for human dry-preserved cell sheets (dry sheets), we compared the intracellular FGF2 levels in seven types of cells reported as cell sheets for clinical use or preclinical studies. FGF2 levels were high in mesenchymal cells, including human oral fibroblasts (HOFs) and human dermal fibroblasts (HDFs), human dental pulp stem cells (DPSCs), and human mesenchymal stem cells (MSCs); in contrast, FGF2 levels in human umbilical vascular endothelial cells (HUVECs), human skeletal muscle myoblasts (SkMMs), and human epidermal keratinocytes (HEKs) were remarkably low, approximately 25% those in fibroblasts. In addition, we prepared dry sheets from HOFs, DPSCs, and MSCs, and analyzed the growth factors released from each dry sheet upon rehydration. High levels of FGF2, HGF, and VEGF were detected in the eluate prepared by immersing each dry sheet. In particular, FGF2 and HGF were the most abundant in HOFs. An in vitro cell proliferation assay showed that these eluates significantly enhanced HUVEC proliferation compared to control cells. Furthermore, cells incubated with HOF eluate showed significantly higher cell proliferation than cells incubated with DPSC and MSC eluates. However, this proliferative response was significantly blocked by FGF2-neutralizing antibodies. These results demonstrate that growth factors released from human dry sheets have physiological activity and that this activity is mainly mediated by the effect of FGF2. Fibroblasts are ideal for the clinical application of dry-preserved cell sheets in humans owing to their high intracellular FGF2 content, fast cell proliferation, ease of handling, availability, and low culture costs, making them the most suitable cell source for regenerative medicine, with FGF2 release as the mechanism of action.

Analysis of mRNA-miRNA interaction network reveals the role of CAFs-derived exosomes in the immune regulation of oral squamous cell carcinoma

BackgroundCancer-associated fibroblasts (CAFs) have significant tumor regulatory functions, and CAFs-derived exosomes (CAFs-Exo) released from CAFs play an important role in the progression of oral squamous cell carcinoma (OSCC). However, a lack of comprehensive molecular biological analysis leaves the regulatory mechanisms of CAFs-Exo in OSCC unclear.MethodsWe used platelet derived growth factor-BB (PDGF-BB) to induce the transformation of human oral mucosa fibroblast (hOMF) into CAFs, and extracted exosomes from the supernatant of CAFs and hOMF. We validated the effect of CAFs-Exo on tumor progression by exosomes co-culture with Cal-27 and tumor-forming in nude mice. The cellular and exosomal transcriptomes were sequenced, and immune regulatory genes were screened and validated using mRNA-miRNA interaction network analysis in combination with publicly available databases.ResultsThe results showed that CAFs-Exo had a stronger ability to promote OSCC proliferation and was associated with immunosuppression. We discovered that the presence of immune-related genes in CAFs-Exo may regulate the expression of PIGR, CD81, UACA, and PTTG1IP in Cal-27 by analyzing CAFs-Exo sequencing data and publicly available TCGA data. This may account for the ability of CAFs-Exo to exert immunomodulation and promote OSCC proliferation.ConclusionsCAFs-Exo was found to be involved in tumor immune regulation through hsa-miR-139-5p, ACTR2 and EIF6, while PIGR, CD81, UACA and PTTG1IP may be potentially effective targets for the treatment of OSCC in the future.

Development of the Follow-Up Human 3D Oral Cancer Model in Cancer Treatment.

As function preservation cancer therapy, targeted radiation therapies have been developed for the quality of life of cancer patients. However, preclinical animal studies evaluating the safety and efficacy of targeted radiation therapy is challenging from the viewpoints of animal welfare and animal protection, as well as the management of animal in radiation-controlled areas under the regulations. We fabricated the human 3D oral cancer model that considers the time axis of the follow up in cancer treatment. Therefore, in this study, the 3D model with human oral cancer cells and normal oral fibroblasts was treated based on clinical protocol. After cancer treatment, the histological findings of the 3D oral cancer model indicated the clinical correlation between tumor response and surrounding normal tissue. This 3D model has potential as a tool for preclinical studies alternative to animal studies.

Cytotoxicity of Impregnated Dental Gingival Retraction Cord Extracts in Immortalized Human Oral Fibroblasts and Keratinocytes in vitro

Cytotoxicity of Impregnated Dental Gingival Retraction Cord Extracts in Immortalized Human Oral Fibroblasts and Keratinocytes in vitro

In Vitro Cell Proliferation and Migration Properties of Oral Mucosal Fibroblasts: A Comparative Study on the Effects of Cord Blood- and Peripheral Blood-Platelet Lysate.

Cord blood-platelet lysate (CB-PL), containing growth factors such as a platelet-derived growth factor, has a similar efficacy to peripheral blood-platelet lysate (PB-PL) in initiating cell growth and differentiation, which makes it a unique alternative to be implemented into oral ulceration healing. This research study aimed to compare the effectiveness of CB-PL and PB-PL in promoting oral wound closure in vitro. Alamar blue assay was used to determine the optimal concentration of CB-PL and PB-PL in enhancing the proliferation of human oral mucosal fibroblasts (HOMF). The percentage of wound closure was measured using the wound-healing assay for CB-PL and PB-PL at the optimal concentration of 1.25% and 0.3125%, respectively. The gene expressions of cell phenotypic makers (Col. I, Col. III, elastin and fibronectin) were determined via qRT-PCR. The concentrations of PDGF-BB were quantified using ELISA. We found that CB-PL was as effective as PB-PL in promoting wound-healing and both PL were more effective compared to the control (CTRL) group in accelerating the cell migration in the wound-healing assay. The gene expressions of Col. III and fibronectin were significantly higher in PB-PL compared to CB-PL. The PDGF-BB concentration of PB-PL was the highest and it decreased after the wound closed on day 3. Therefore, we concluded that PL from both sources can be a beneficial treatment for wound-healing, but PB-PL showed the most promising wound-healing properties in this study.

Antifungal efficiency and cytocompatibility of polymethyl methacrylate modified with zinc dimethacrylate.

Considering the high incidence rates of denture stomatitis, research that providing dental biomaterials with antifungal property are essential for clinical dentistry. The objectives of the present study were to investigate the effect of zinc dimethacrylate (ZDMA) modification on the antifungal and cytotoxic properties, as well as the variance in surface characteristics and other physicochemical properties of polymethyl methacrylate (PMMA) denture base resin. PMMA with various mass fraction of ZDMA (1 wt%, 2.5 wt% and 5 wt%) were prepared for experimental groups, and unmodified PMMA for the control. Fourier-transform infrared spectroscopy (FTIR) was applied for characterization. Thermogravimetric analysis, atomic force microscopy and water contact angle were performed to investigate the thermal stability and surface characteristics (n=5). Antifungal capacities and cytocompatibility were evaluated with Candida albicans (C. albicans) and human oral fibroblasts (HGFs), respectively. Colony-forming unit counting, crystal violet assay, live/dead biofilm staining and scanning electron microscopy observation were performed to assess antifungal effects, and the detection of intracellular reactive oxygen species production was applied to explore the possible antimicrobial mechanism. Finally, the cytotoxicity of ZDMA modified PMMA resin was evaluated by the 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and live/dead double staining. The FTIR analyses confirmed some variation in chemical bonding and physical blend of the composites. Incorporation of ZDMA significantly enhanced the thermal stability and hydrophilicity compared with unmodified PMMA (p < 0.05). The surface roughness increased with the addition of ZDMA while remained below the suggested threshold (≤ 0.2 µm). The antifungal activity significantly improved with ZDMA incorporation, and cytocompatibility assays indicated no obvious cytotoxicity on HGFs. In the present study, the ZDMA mass fraction up to 5 wt% in PMMA performed better thermal stability, and an increase in surface roughness and hydrophilicity without enhancing microbial adhesion. Moreover, the ZDMA modified PMMA showed effective antifungal activity without inducing any cellular side effects.

Reduced Fibroblast Activation on Electrospun Polycaprolactone Scaffolds.

In vivo, quiescent fibroblasts reside in three-dimensional connective tissues and are activated in response to tissue injury before proliferating rapidly and becoming migratory and contractile myofibroblasts. When deregulated, chronic activation drives fibrotic disease. Fibroblasts cultured on stiff 2D surfaces display a partially activated phenotype, whilst many 3D environments limit fibroblast activation. Cell mechanotransduction, spreading, polarity, and integrin expression are controlled by material mechanical properties and micro-architecture. Between 3D culture systems, these features are highly variable, and the challenge of controlling individual properties without altering others has led to an inconsistent picture of fibroblast behaviour. Electrospinning offers greater control of mechanical properties and microarchitecture making it a valuable model to study fibroblast activation behaviour in vitro. Here, we present a comprehensive characterisation of the activation traits of human oral fibroblasts grown on a microfibrous scaffold composed of electrospun polycaprolactone. After over 7 days in the culture, we observed a reduction in proliferation rates compared to cells cultured in 2D, with low KI67 expression and no evidence of cellular senescence. A-SMA mRNA levels fell, and the expression of ECM protein-coding genes also decreased. Electrospun fibrous scaffolds, therefore, represent a tuneable platform to investigate the mechanisms of fibroblast activation and their roles in fibrotic disease.

Histocompatibility of Light-Curing Composites Used in Pediatric Dentistry in Human Oral Fibroblast Cells.

Tooth-colored composites are used to repair caries lesions and other dental defects, particularly in anterior regions in children. Although a wide range of composites is using, little attention has been paid to the important indicators such as biological profiles or products released from these materials. The current study aimed to compare the histocompatibility and cytotoxicity of light-curing resin used to repair children's teeth with different brands (3M, DenFil, and Opallis) in curing times of 20 and 40 seconds in human oral fibroblast cells (HGF1). In this in vitro study, Three types of flow composites (3M, Opallis, and DenFil), all at A2 shade, were used. The composites were at 4×2mm with separate exposure times of 20 and 40 seconds. MTT test was used to determine the cytotoxicity of composites on oral fibroblast cells. This test is based on the conversion of tetrazolium bromide to a purple compound known as formazan that its color intensity can be evaluated using the ELISA. The higher intensity of the color reveals the higher survival rate of the mitochondria, which indicates less toxicity. One-way variance analysis and unpaired t-test were used to compare the cytotoxicity of each brand in two conditions of 20 and 40 seconds of curing. Statistical significance was considered when p< 0.05. 3M and Opallis composites were significantly reduced vitality of cells compared to control group in both 20s and 40s curing status. While DenFil brand did not show marked cytotoxicity. In each brand, there are no significant deference between 20s and 40s curing times. Histocompatibility depends on the type of composite resin. In the current study, DenFil brand showed the highest histocompatibility, followed by 3M and Opallis.