Increased Fibroblast Proliferation Research Articles

Zinc oxide nanoparticles induce human tenon fibroblast apoptosis through reactive oxygen species and caspase signaling pathway

Glaucoma is the leading cause of irreversible blindness in the world and trabeculectomy remains still the most commonly performed filtration surgery. Failure of trabeculectomy is due to the formation of scarring, which is associated with the increased fibroblast proliferation, activation, and collagen deposition at the site of the drainage channel with subconjunctival fibrosis. Our previous study has revealed that zinc oxide (ZnO) nanoparticles could efficiently decrease the expressions of TGF-β1 and inhibit fibroblast-mediated collagen lattice contraction. However, the mechanism underlying ZnO nanoparticle-induced fibroblast apoptosis is still unclear. In the present study, we investigated the effect of ZnO nanoparticles on the reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm) in human Tenon fibroblasts (HTFs). Moreover, we also explored the influence of ZnO nanoparticles on the expression of Caspase-3, Caspase-9, apoptotic protease-activating factor-1 (Apaf-1), fibroblast-specific protein-1 (FSP-1), collagen III, and E-cadherin. The results indicated that ZnO nanoparticles markedly inhibit HTFs viability and decrease the Δψm in a concentration-dependent pattern. Exposure of HTFs to ZnO nanoparticles could also induce the elevated Caspase-3, Caspase-9, and Apaf-1 expression, decrease the levels of FSP-1, collagen III, and E-cadherin expression, leading to HTFs apoptosis. Our results suggested that elevated ROS and activated Caspase signaling play a fundamental role in ZnO nanoparticle-induced HTFs apoptosis.

Benefits of Gotu Kola Leaf in Responding to Prevent The Form of Keloid in The Wound

Keloids are a type of scar, which is the result of overgrowth of dense fibrous tissue that usually develops after healing a skin injury. Keloids occur due to prolonged inflammatory phases in the wound, resulting in increased fibroblast activity and continuous extracellular matrix formation. Gotu kola (Centella asiatica) leaves are effective in the treatment of wounds because they contain Triterpene components such as asiatic acid, madecassic acid, asiaticoside and madecassoside. The purpose of this literature review is to report the latest scientific findings on the role of gotu kola leaf content in preventing keloid formation by increasing fibroblast proliferation, increasing collagen synthesis and mucopolysaccharide acids, increasing intracellular fibronectin which then significantly increases the tensile strength of newly formed skin, inhibiting the phases of the formation of skin, inflammation of the keloid scar tissue. The method used in this article is article search through the NCBI database and Google Scholar. The year of publication of library resources is from 1999 to 2018 with 20 sources of literature. The theme collected was related to the Centella asiatica leaf content towards the wound phase to prevent keloid formation. The results of the synthesis of articles that have been found are gotu kola leaves can inhibit the formation of keloids in wounds.
 Keywords: gotu kola leaf, keloid, wound

Healing of Skin Wounds in Rats: Influence of Low-Level Laser Therapy, Alone or Combined with Natural Products

Background: A great variety of natural products have been evaluated for the skin wound healing due to anti-inflammatory, antioxidant and antibacterial activities, and procollagen synthesis, of which may be mentioned the honey and propolis. In turn, low-level laser therapy (LLLT) is considered an important tool in the treatment of skin wounds, because of ability to raise the ATP production, to stimulate the microcirculation, and formation of new blood vessels. Therefore,this study aimed to assess the influence of LLLT, alone or combined with natural products, in the healing of excisional skin wounds in rats. Materials, Methods & Results: One hundred twenty-six male Wistar rats, aged 14 weeks, were randomly divided into seven groups (n = 18 per group) according to wound treatments: G1 (negative control): 0.9% saline solution; G2 (positive control): allantoin + zinc oxide ointment; G3: LLLT; G4: mixture of honey + 5% propolis hydroalcoholic solution; G5: LLLT + honey + hydroalcoholic solution of 5% propolis; G6: LLLT + honey in natura; G7: LLLT + 5% propolis hydroalcoholic solution. Six rats from each group were euthanized on the 7th, 14th, and 21th postoperative days. Macroscopic and histological evaluations of wound healing were performed. A decrease in wound area occurred in all groups, but in general G4 had the higher reduction and G1 and G3 had the lower reductions. These findings corresponded to the percentage of wound contraction, since the largest contraction was G4 and the lowest contractions were G1 and G3. Histological analysis showed no significant difference among groups on the 7th postoperative day. On postoperative day 14, a significant decrease in hemorrhage occurred between G1xG4, G1xG5 and G1xG7, and a significant decrease in congestion between G1xG7 and G2xG5. In addition, intensity of the inflammatory infiltrate showed a significant reduction between G1xG7, G2xG7 and G3xG7. On postoperative day 21, a significant changed from mixed to mononuclear inflammatory infiltrate was observed between G1xG3, G1xG4, G1xG6, G1xG7, G2xG3, and G2xG6. Regarding organization of fibroblasts, there were differences between G1xG6 and G1xG7. Discussion: Spite the red laser has been associated to in increase fibroblast proliferation, G3 showed inadequate decrease in wound area and percentage of contraction. Histological examination showed no differences among groups in the inflammatory phase. However, in the proliferative and remodeling phases around days 14 and 21 were found some differences among groups, which may be related to the actions of the natural products, or laser. Studies have shown that honey acts as a repair and anti-inflammatory agent in skin woundthat may be associated to lower hemorrhage in G4 and G4 compared to G1 on day 14. Propolis is related to reduction of free radicals and increasing the amount of collagen, which may have contributed for quality improvement of the inflammatory infiltrate in G4 and G7 compared to G1, and fibroblast organization in G7 at postoperative day 21. In conclusion, macroscopically the mixture of honey + 5% propolis hydroalcoholic solution was the most effective in reducing wound area and increasing wound contraction. However, based on the parameters evaluated histologically, may be highlighted the evolution of the treatments with LLLT + honey, as well as LLLT + 5% propolis hydroalcoholic solution. Keywords: honey, propolis, biomodulation, wound, healing.

CD26 upregulates proliferation and invasion in keloid fibroblasts through an IGF-1-induced PI3K/AKT/mTOR pathway.

BackgroundKeloid is a fibrotic dermal disease characterized by an abnormal increase in fibroblast proliferation and invasion. These pathological behaviours may be related to the heterogeneity of keloid fibroblasts (KFs); however, because of a lack of effective biomarkers for KFs it is difficult to study the underlying mechanism. Our previous studies revealed that the expansion of CD26+ KFs was responsible for increased keloid proliferation and invasion capabilities; the intrinsic relationship and mechanism between CD26 and keloid is therefore worthy of further investigation. The aim of this study was to explore molecular mechanisms in the process of CD26 upregulated KFs proliferation and invasion abilities, and provide more evidence for CD26 as an effective biomarker of keloid and a new clinical therapeutic target.MethodsFlow cytometry was performed to isolate CD26+/CD26− fibroblasts from KFs and normal fibroblasts. To generate stably silenced KFs for CD26 and insulin-like growth factor-1 receptor (IGF-1R), lentiviral particles encoding shRNA targeting CD26 and IGF-1R were used for transfection. Cell proliferations were analysed by cell counting kit-8 assay and 5-ethynyl-2′-deoxyuridine (EdU) incorporation assay. Scratching assay and transwell assay were used to assess cell migration and invasion abilities. To further quantify the regulatory role of CD26 expression in the relevant signalling pathway, RT-qPCR, western blot, ELISA, PI3K activity assay and immunofluorescence were used.ResultsAberrant expression of CD26 in KFs was proven to be associated with increased proliferation and invasion of KFs. Furthermore, the role of the IGF-1/IGF-1 receptor axis was also studied in CD26 and was found to upregulate KF proliferation and invasion. The PI3K/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway was shown to affect CD26-regulated KF proliferation and invasion by increasing phosphorylation levels of S6 kinase and 4E-binding protein.ConclusionsCD26 can be the effective biomarker for KFs, and its expression is closely related to proliferation and invasion in keloids through the IGF-1-induced PI3K/AKT/mTOR pathway. This work provides a novel perspective on the pathological mechanisms affecting KFs and therapeutic strategies against keloids.

The effect of purple sweet potato (Ipomea batatas L.) extract gel toward TNF-α expression and skin collagen density on rats (Rattus norvegicus) with open wound models

Open wound is a damage to the skin or tissues under the skin that can be caused by sharp objects or fights between animals. Purple sweet potato (Ipomea batatas L) contains some bioactive substances, such as anthocyanin and tannin, which help to prevent the prolongation of the inflammatory phase and increase fibroblast proliferation. The aim of this research was to determine the effect of purple sweet potato extract gel toward the expression of TNF-α and skin collagen density on rats with open wound model. The open wound was made circular using a modified Morton method. This study used a completely randomized design with 20 rats, Wistar strain aged 12 weeks, weighing 150-200 g, divided into 5 groups with 4 replicates. TNF-α expression observed in immunohistochemistry method and analyzed using one-way ANOVA statistical test, followed by Honestly Significant Difference Test. The density of collagen with Masson Trichome staining was analyzed by the Kruskal-Wallis nonparametric statistic test with Mann-Whitney-U test. The results showed that the purple sweet potato extract as a therapy for open wounds was significantly reduce the TNF-α expression and increase collagen density. The conclusion is that the purple sweet potato extract with 30% concentration is the most effective therapy in increasing collagen density but not very effective in reducing TNF-α expression.

Stromal Features of the Primary Tumor Are Not Prognostic in Genetically Engineered Mice of Pancreatic Cancer.

The KrasG12D/+;LSL-Trp53R172H/+;Pdx-1-Cre (KPC) mouse model is frequently employed for preclinical therapeutic testing, in particular in regard to antistromal therapies. Here, we investigate the prognostic implications of histopathological features that may guide preclinical trial design. Pancreatic tumor tissue from n = 46 KPC mice was quantitatively analyzed using immunohistochemistry and co-immunofluorescence for proliferation (Ki67), mitotic rate (phospho-Histone 3, PHH3), apoptosis (cleaved caspase-3, CC3), collagen content, secreted protein acidic and rich in cysteine (SPARC), hyaluronic acid (HA), and α-smooth muscle actin (α-SMA). Furthermore, mean vessel density (MVD), mean lumen area (MLA), grading, activated stroma index (ASI), and fibroblast-proliferation rate (α-SMA/Ki67) were assessed. Univariate analysis using the Kaplan–Meier estimator and Cox regression model for continuous variables did not show association between survival and any of the analyzed parameters. Spearman correlation demonstrated that desmoplasia was inversely correlated with differentiated tumor grade (ρ = −0.84). Ki67 and PHH3 synergized as proliferation markers (ρ = 0.54), while SPARC expression was positively correlated with HA content (ρ = 0.37). MVD and MLA were correlated with each other (ρ = 0.31), while MLA positively correlated with CC3 (ρ = 0.45). Additionally, increased MVD was correlated with increased fibroblast proliferation rate (α-SMA + Ki67; ρ = 0.36). Our pilot study provides evidence that individual histopathological parameters of the primary tumor of KPC mice are not associated with survival, and may hint at the importance of systemic tumor-related effects such as cachexia.

A novel bilayer zein/MMT nanocomposite incorporated with H. perforatum oil for wound healing.

Recently, layered structures composed of nanofibers have gained attention as a novel material to mimic skin tissue in wound healing applications. The aim of this study is to develop a novel hybrid bilayer material composed of zein based composite film and nanofiber layers as a wound dressing material. The upper layer was composed of H. perforatum oil incorporated zein film including MMT and the bottom layer was comprised of 3D electrospun zein/MMT nanofibers to induce wound healing with the controlled release of H. perforatum oil. The bilayer composites were characterized in terms of mechanical test, WVP, water uptake and surface wettability. Antimicrobial activity of the wound dressings against microorganisms were investigated by disc diffusion method. In vitro cytotoxicity of monolayer film and bilayer structure was performed using WST-1 assay on HS2 keratinocyte and 3T3 cell lines. Results indicated that the prepared monolayer films showed appropriate mechanical and gas barrier properties and surface wettability for wound healing. Controlled release of H. perforatum oil was obtained from fabricated membranes up to 48 h. Bilayer membranes showed antimicrobial activity against E. coli, S. aureus, and C. albicans and did not show any toxic effect on NIH3T3 mouse fibroblast and HS2 keratinocyte cell lines. In vitro scratch assay results indicated that H. perforatum oil had a wound healing effect by inducing fibroblast migration. The proliferation study supported these results by increasing fibroblast proliferation on H. perforatum oil loaded bilayer membranes.

Tissue remodelling in chronic rhinosinusitis - review of literature.

CRS is a process involving a number of adverse changes in the mucosa of the paranasal sinuses and nasal polyps, e.g. increased fibroblast proliferation, angiogenesis, increased formation of fibrous tissue (subepithelial fibrosis) and tissue destruction. There are biomarkers whose levels can be increased in chronic inflammation of the paranasal sinuses: peripheral blood eosinophilia, IgE immunoglobulin, cytokines - IL-4, IL-5, IL-13, IL-25, IL-33, periostin, P-glycoprotein, CXCL-12, CXCL-13, INF-Υ, TNFα, TGFβ1, albumins, eotaxin. These biomarkers are not pathognomonic for CRS. The concentration of biomarkers is also increased in bronchial asthma and atopic dermatitis. The TGFβ, in particular, the β1 subunit, was identified as the main factor involved in the remodeling of tissue stroma. In conjunction with the continuous improvement of tissue testing methods, it is advisable to search for new factors that will more accurately allow the assessment of tissue remodeling in the chronic processes of paranasal sinuses.

Effect of Platelet-rich Plasma on Implant Bone Defects in Rabbits Through the FAK/PI3K/AKT Signaling Pathway.

Platelet-rich plasma (PRP) has been shown to be a beneficial growth factor for bone tissue healing and is used in implantology. The aim of this study was to investigate the effects of PRP on bone defects in rabbits. Twenty rabbits were used to establish the implant bone defect model in this study. An intrabony defect (5mm × 5mm × 3mm) was created in alveolar bone in the lower jar of each rabbit. The wound was treated with PRP. The expression of platelet-derived growth factor BB (PDGFBB) was assessed by enzyme-linked immunosorbent assay (ELISA). Focal adhesion kinase (FAK) and related phosphatidylinositol 3-kinase (PI3K)/AKT (protein kinase B) levels were measured by Western blot. The results show that PRP could significantly improve the bone healing process when compared with control, and 10% PRP could markedly increase fibroblast proliferation 48-h post treatment. PDGFBB was higher in the PRP group than that in the control group. PRP treatment also could elevate the phosphorylation of FAK and PI3K/AKT, although the inhibitor of PDGFR could reverse this trend. These results suggest that PRP treatment improves the bone healing process through the FAK/PI3K/AKT pathway.

The Role of MicroRNA-21 in Venous Neointimal Hyperplasia: Implications for Targeting miR-21 for VNH Treatment.

The molecular mechanism of hemodialysis access arteriovenous fistula (AVF) failure due to venous neointimal hyperplasia (VNH) is not known. The role of microRNA-21 (miR-21) in VNH associated with AVF failure was investigated by performing in vivo and in vitro experiments. In situ hybridization results revealed that miR-21 expression increased and was associated with fibroblasts in failed AVFs from patients. In a murine AVF model, qRT-PCR gene expression results showed a significant increase in miR-21 and a decrease in miR-21 target genes in graft veins (GVs) compared to contralateral veins in mouse AVF. miR-21 knockdown in GVs was performed using a lentivirus-mediated small hairpin RNA (shRNA), and this improved AVF patency with a decrease in neointima compared to control GVs. Moreover, loss of miR-21 in GVs significantly decreased the Tgfβ1, Col-Ia, and Col-Iva genes. Immunohistochemistry demonstrated a significant decrease in myofibroblasts and proliferation with an increase in terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining in miR-21-knockdown vessels, along with a decrease in hypoxia-inducible factor-1 alpha (HIF-1α) and phospho-SMAD2 (pSMAD-2) and phospho-SMAD3 (pSMAD-3) and an increase in phosphatase and tensin homolog (PTEN) staining. Hypoxic fibroblast knockdown for miR-21 showed a significant decrease in Tgfβ-1 expression and pSMAD-2 and -3 levels and a decrease in myofibroblasts. These results indicate that miR-21 upregulation causes VNH formation by fibroblast-to-myofibroblast differentiation.

MicroRNA-1-Mediated Inhibition of Cardiac Fibroblast Proliferation Through Targeting Cyclin D2 and CDK6.

MicroRNA-1 (miRNA-1) has been long viewed as a muscle-specific miRNA and plays a critical role in myocardium and cardiomyocytes by controlling myocyte growth and rhythm. We identified that miRNA-1 is expressed in cardiac fibroblasts, which are one of the major non-muscle cell types in myocardium and are responsible for cardiac fibrosis in pathological conditions. In this study, we aimed to investigate the effect and mechanism of action of miRNA-1 on cardiac fibroblast proliferation. Subcutaneous angiotensin II (Ang II) infusion via osmotic minipumps for 4 weeks was used to induce myocardial interstitial fibrosis in male Sprague-Dawley rats. MiRNA-1 expression was significantly down-regulated by 68% in freshly isolated ventricular fibroblasts from Ang II-infused rats than that from control rats. Similar results were obtained in adult rat ventricular fibroblasts that were stimulated in culture by Ang II or TGFβ for 48 h. Functionally, overexpression of miRNA-1 inhibited fibroblast proliferation, whereas knockdown of endogenous miRNA-1 increased fibroblast proliferation. We then identified and validated cyclin D2 and cyclin-dependent kinase 6 (CDK6) as direct targets of miRNA-1 in cardiac fibroblasts using biochemical assays. Moreover, we showed that the inhibitory effects of miRNA-1 on cardiac fibroblast proliferation can be blunted by overexpression of its target, cyclin D2. In conclusion, our findings demonstrate miRNA-1 expression and regulation in adult ventricular fibroblasts, where it acts as a novel negative regulator of adult cardiac fibroblast proliferation that is at least partially mediated by direct targeting of two cell cycle regulators. Our results expand the understanding of the regulatory roles of miRNA-1 in cardiac cells (i.e., from myocytes to a major non-muscle cells in the heart).

Decreased HSP70 expression on serum exosomes contributes to cardiac fibrosis during senescence.

Aging is now considered as an independent risk factor for cardiac fibrosis. However, the mechanisms underlying aging-related cardiac fibrosis remain unknown. Here, we examine the role of serum exosomes in this process. Experiments were conducted using 6-week-old or 24-week-old male Sprague-Dawley (SD) rats. Cardiac sections were treated with Masson's trichrome stain to evaluate fibrosis. Exosomes were isolated from the serum, characterized and quantified using Western blot, electron microscopy, and qNano analysis, and co-cultured with transforming growth factor-β1 (TGF-β1)-induced primary cardiac fibroblasts (CF). Co-cultures were also carried out in the presence of a hot shock protein 70 (HSP70) inhibitor (gefitinib) or inducer (geranylgeranylacetone) to evaluate the role of HSP70 in cardiac fibrosis. Cardiac fibrosis as well as serum exosomes levels were increased during senescence. We observed an increase in fibroblast proliferation and myofibroblast differentiation when CF were co-cultured with exosomes from old rats, compared to those from young mice. Observing a decrease in surface HSP70 expression on the exosomes derived from old rats, we tested the effects of HSP70 inhibition or overexpression on the CF co-cultures. HSP70 inhibition increased fibroblast proliferation and myofibroblast differentiation in CF co-cultures containing exosomes from the young rats, while HSP70 overexpression attenuated fibroblast proliferation and myofibroblast differentiation in CF co-cultures containing exosomes from the old rats. Using an animal model of cardiac fibrosis, we show a decrease in HSP70 expression on the exosomal surface with aging, which may contribute to cardiac fibrosis.

Impact of Er:YAG laser on wound healing following nonsurgical therapy: A pilot study.

The purpose is to examine early wound healing through histological analysis by characterizing connective tissue distribution and organization in the treated periodontium following nonsurgical therapy. Periodontal disease is a multifactorial pathological process that leads to the loss of the surrounding periodontium. Traditional periodontal therapies have proven beneficial in halting the progression of disease. The aim of this study is to investigate early wound healing in periodontal patients following hand/ultrasonic instrumentation alone, erbium‐doped yttrium aluminum garnet laser instrumentation alone, or a combination of hand/ultrasonic instrumentation and Er:YAG laser instrumentation for the nonsurgical treatment of periodontitis by histologic evaluation. Twenty‐one patients were randomized to receive nonsurgical therapy for the treatment of chronic periodontitis with three modalities prior to surgical therapy. Baseline clinical measurements were obtained prior to treatment. Wound healing was assessed by obtaining an otherwise discarded tissue sample following nonsurgical therapy of the selected study site. Samples were obtained at 2 or 6 weeks following initial therapy with a step‐back incision and fixated for histological and immunohistochemical analysis. There were minimal between‐group differences in the amount of collagen distribution when analyzing the Mallory–Heidenhain Azan trichrome, Picrosirus Red stain, and proliferating cell nuclear antigen at both time points. Descriptive analysis of baseline measurements showed no differences in probing depth change, bleeding on probing, and clinical attachment level following initial therapy between the three treatment groups at 2 or 6 weeks. Each treatment modality was effective in treating moderate to severe chronic periodontitis; however, the results of this study are inconclusive regarding superiority of any one treatment approach from a histologic and immunohistochemical perspective. Based on this assessment, there was increased fibroblast proliferation and collagen maturation between the 2‐ and 6‐week time point after treatment in all treatment groups, with few apparent differences between treatment groups. This pilot study qualitatively evaluated early wound healing in periodontal patients following non surgical therapy with various treatment modalities. When comparing descriptive outcomes of Er:YAG laser therapy and hand/ultrasonic instrumentation there were minimal differences in collagen distribution and density between groups. The evaluated modalities were each effective treating periodontal patients with non surgical therapy.

Secretory Inositol Polyphosphate 4-Phosphatase Protects against Airway Inflammation and Remodeling.

The asthma candidate gene inositol polyphosphate 4-phosphatase type I A (INPP4A) is a lipid phosphatase that negatively regulates the PI3K/Akt pathway. Destabilizing genetic variants of INPP4A increase the risk of asthma, and lung-specific INPP4A knockdown induces asthma-like features. INPP4A is known to localize intracellularly, and its extracellular presence has not been reported yet. Here we show for the first time that INPP4A is secreted by airway epithelial cells and that extracellular INPP4A critically inhibits airway inflammation and remodeling. INPP4A was present in blood and BAL fluid, and this extracellular INPP4A was reduced in patients with asthma and mice with allergic airway inflammation. In both naive mice and mice with allergic airway inflammation, antibody-mediated neutralization of extracellular INPP4A potentiated PI3K/Akt signaling and induced airway hyperresponsiveness, with prominent airway remodeling, subepithelial fibroblast proliferation, and collagen deposition. The link between extracellular INPP4A and fibroblasts was investigated in vitro. Cultured airway epithelial cells secreted enzymatically active INPP4A in extracellular vesicles and in a free form. Extracellular vesicle-mediated transfer of labeled INPP4A, from epithelial cells to fibroblasts, was observed. Inhibition of such transfer by anti-INPP4A antibody increased fibroblast proliferation. We propose that secretory INPP4A is a novel "paracrine" layer of the intricate regulation of lung homeostasis, by which airway epithelium dampens PI3K/Akt signaling in inflammatory cells or local fibroblasts, thereby limiting inflammation and remodeling.

Is leukotriene B4 one of the keloid marker? a fibroblast keloid study

Keloid pathogenesis occurs due to the duration of prolonged inflammatory phase and increased production of various growth factors such as TGF-β1 which may cause increasing fibroblast proliferation and collagen synthesis. Existence of one of the chemical mediators released during inflammation, leukotriene B4 (LTB4), in keloid pathogenesis specifically in the phases of inflammation and proliferation, is still unclear. The purpose of the study is to analyze the levels of LTB4 in keloid. Methods: Fibroblast culture that was done by explanting keloid and normal skin of a keloid patient. Measurement of LTB4 on keloid and normal fibroblast was done by Elisa method. This experiment was run in triplicate. Statistical test was conducted by t test for the unpaired data and Anova test. The experiment was done at cell culture laboratory of Medical Faculty of Padjadjaran University Bandung. Levels of LTB4 in keloid fibroblast was higher than that of normal skin fibroblast (mean 23143.27 vs 18191.85 pg/ml; p<0.05). Conclusion, increased LTB4 levels in keloid fibroblast showed the existence of LTB4 role in the prolonged inflammatory process in keloid pathogenesis.

Study of hybrid alginate/gelatin hydrogel-incorporated niosomal Aloe vera capable of sustained release of Aloe vera as potential skin wound dressing

Nowadays, wound dressings serve as advanced skin products, which mainly aim to accelerate the wound healing process. The wound dressings with a potential of localized and prolonged release of therapeutic agents have attracted tremendous attention. In this study, synthesis of a sustained release of niosomal Aloe vera (AV) loaded in alginate/gelatin (AG) hybrid hydrogel is aimed at improving skin regeneration as wound dressing. For this purpose, AV-loaded niosomes are synthesized and incorporated in the hybrid AG hydrogel. The size and polydispersity index (PdI) of niosomes, AV entrapment efficacy and AV in vitro release are characterized. In addition, the hydrogel characteristic, such as swelling ratio, degradation behavior and mechanical property, are studied. MTT assay is utilized to evaluate the effect of AV incorporation and release on the proliferation of fibroblast cells. Results demonstrate that size, PdI and EE% of AV-loaded niosomes are 270.080 nm, 0.108 and 42.039 ± 4.090%, respectively, and in vitro release of AV is about 20% after 7 days. AG hybrid hydrogel loaded by niosomal AV shows an extended sustained release manner, where its swelling ratio and percentage of degradation are 60 wt% after 72 h and 70 wt% after 6 days, respectively. The average Young’s modulus of the hybrid hydrogel is measured around 12.64 ± 1.3 kPa, which seems suitable as a wound dressing. Finally, MTT assay confesses an increased fibroblast proliferation in the presence of AV, particularly in the niosomal AV-loaded hydrogel. Concludingly, alginate/gelation hybrid hydrogel incorporated with niosomal AV, with sustained release potential can be suggested as a promising candidate for wound dressing applications.

An in vitro study of scarring formation mediated by human Tenon fibroblasts: Effect of Y-27632, a Rho kinase inhibitor.

Scar formation is the most common cause for failure of glaucoma filtration surgery because of increased fibroblast proliferation and activation. We have now examined the effect of Y‐27632, a Rho‐associated protein kinase (ROCK) inhibitor, on postsurgical scarring formation in human Tenon fibroblasts (HTFs). Collagen gel contraction assay was used to compare contractility activity of Y‐27632 with several antiglaucoma drugs. Immunofluorescence and western blotting were used to examine expression of scar formation–related factors. We found that Y‐27632 inhibited collagen gel contraction, as well as α‐smooth muscle actin and vimentin expression; these were promoted by treatment with latanoprost, timolol, or transforming growth factor (TGF)–β. To investigate the effect of Y‐27632 in postsurgical scarring, we mimicked TGF‐β secretion by stimulating HTFs with TGF‐β prior to Y‐27632 treatment. HTFs cultured in the presence of TGF‐β significantly increased gel contraction. In contrast, when HTFs were treated with 10μM Y‐27632, contraction was significantly inhibited. Furthermore, Y‐27632 reduced TGF‐β–induced phosphorylation of mitogen‐activated protein kinase signalling. These results suggest that ROCK inhibitors may inhibit fibrosis by inhibiting transdifferentiation of Tenon fibroblasts into myofibroblasts and by inhibiting TGF‐β signalling after surgery through mitogen‐activated protein kinase pathway suppression. These results implicate that ROCK inhibitors may improve outcomes after filtering surgery with a potential antiscarring effect, while latanoprost and timolol may induce fibrosis.Significance of the studyScar formation is the primary cause of failure after glaucoma filtration surgery. A ROCK inhibitor, Y‐27632, has been introduced as a novel potential antiglaucoma treatment to reduce intraocular pressure. The aim of our study was to elucidate the effect of Y‐27632 on scarring formation after glaucoma filtration surgery, in direct comparison with other antiglaucoma drugs. Our findings thus suggested that Y‐27632 may inhibit fibrosis and improve outcome after glaucoma filtration surgery through inhibition of transdifferentiation of Tenon fibroblasts into myofibroblasts, and the TGF‐β and MAPK signalling after surgery, while latanoprost and timolol may induce fibrosis.

The Effects of human amniotic fluid on periodontal ligament fibroblast cell viability, proliferation, and cytokine/growth factor expression

Background: The importance of the amniotic fluid (AF) to the fetus is clear. However, very few studies have been published to examine the potential uses of this fluid in various areas such as tissue regeneration. AF contains epidermal growth factor, transforming growth factor-alpha, transforming growth factor beta-1, insulin-like growth factor-I, erythropoietin and granulocyte colony-stimulating factor, as well as hyaluronic acid and hyaluronic acid-stimulating factor. Previous studies suggest that AF can increase fibroblast proliferation and chemotaxis, and decrease apoptosis as well as promote wound healing. Furthermore, evidence showed that human AF inhibits hyaluronidase, elastase, and cathepsin. The current study examined the effects of human AF on periodontal ligament fibroblasts (PDLF) in terms of cell toxicity, cell proliferation, and cytokine/growth factor expression. Materials and Methods: Cytotoxicity of AF on PDLF was determined using lactate dehydrogenase assays. PDLF proliferation was determined using water-soluble tetrazolium-1 assays. Cytokine/growth factor expression was determined on AF-treated PDLF, AF alone, and PDLF alone utilizing protein arrays. Results: Human AF at 10% and below did not affect cell growth and was not toxic. AF-treated PDLF cells showed a decrease in cytokine/growth factor levels compared to the sum of cytokine/growth factor levels in AF only and cells only for 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased. Conclusion: Human AF at the examined concentrations was not toxic to PDLF cells and did not influence their proliferation. In addition, AF (10%) caused a decrease in the total protein levels of cytokines/growth factors expressed in 39 of the 80 proteins examined (48.8%). Of the 39 examined cytokines, 20 inflammatory cytokines, 11 cell cycle cytokines, 1 anti-inflammatory cytokine, and 7 other cytokines were decreased.

The Effect of Aloe Vera Extract to Extensive Lesion and Expression of Transforming Growth Factor (TGF-β) on Alkaline Chemical Trauma Cornea Model

Abstract
 Background: Chemical trauma to the cornea is an emergency condition of the eye that requires early diagnosis and good treatment. Alkaline have ability to saponify fatty acids in cells and cell membranes which can make penetration into the stroma and destroy proteoglycans and collagen in cells. Aloe vera (AV) contains several active substances that are reported to have anti-inflammatory, immunomodulatory, and wound healing effects. AV has been reported to accelerate the healing process of corneal epithelial defects by increasing fibroblast proliferation, collagen production and growth factor production. This study aims to determine the difference between the effect of aloe vera extract with a concentration of 10%, 20%, 40% and BSS on the healing of extensive corneal lesions in white wistar rats alkaline trauma models.
 Method: This study was an experimental study with a pre and posttest only with control group design in vivo approach to 30 Wistar white rats which were divided into 5 treatment groups for 3 days. Comparative analysis of effectiveness using the ANNOVA test or the Kruskal Wallis test and continued by the post hoc test.
 Results: Based on the one way ANOVA test there was a statistically significant difference in effectiveness between the five treatment groups on the percentage of corneal wound healing area and TGF-β expression with an assessment of p = 0,000 each. The administration of alloevera (AV) concentration of 20% had a significant difference in percentage of healing of corneal lesions and TGF-β expression compared with other treatment groups with p = 0,000 each. Large differences in the area of corneal lesions in the 40% AV group were -0.45 in the BBS group, 0.146 in the 10% AV group, 0.493 in the 20% AV group. The difference in the AV group 10% was 0.30 in the BBS group, -064 in the AV group 20%, and -0.14 in the AV group 40%. However, TGFβ expression in the normal control group that did not receive treatment was 54.94 (53.21-56-12). TGFβ levels in the BSS group were 10.44, the 10% aloe vera group was 25.43, 47.99 for the 20% aloe vera group and 37.95 for the 40% aloe vera group.
 Conclusion: There is a difference between the effect of aloe vera extract with concentrations of 10%, 20%, 40% and BSS on the extensive healing of corneal lesions in white wistar rats with alkaline chemical trauma models.

MiR-203 regulates keloid fibroblast proliferation, invasion, and extracellular matrix expression by targeting EGR1 and FGF2.

Keloid is a fibrous benign tumor of the skin caused by increased fibroblast proliferation and overproduction of extracellular matrix (ECM) in the dermis. Several miRNAs exhibit critical roles in regulating keloid development. This is study aimed to investigate the effects and mechanisms of miR-203 in keloid fibroblasts. The miR-203 expression was detected by qRT-PCR; The cell viability was measured by MTT assay; The cell proliferation was measured by BrdU assay; The cell invasion was measured by Transwell assay; The protein expression was detected by Western blot; The target relationship between miR-203 and mRNA was measured by dual-luciferase assay. We found that miR-203 was significantly downregulated in both keloid tissues and keloid fibroblasts from keloid patients. MiR-203 overexpression in vitro led to a significant decrease of proliferation, invasion, and ECM production in keloid fibroblasts, whereas miR-203 inhibition induced the opposite results. A dual-luciferase reporter assay identified early growth response 1 (EGR1) and fibroblast growth factor 2 (FGF2) as targets of miR-203. EGR1 and FGF2 were overexpressed in keloid fibroblasts and negatively regulated by miR-203. Furthermore, overexpression of EGR1 and FGF2 partially attenuated the suppressive effect of miR-203 on the proliferation, invasion, and ECM production of keloid fibroblasts. In conclusion, we demonstrated for the first time that miR-203 decreased the proliferation, invasion, and ECM production of keloid fibroblasts by repressing EGR1 and FGF2 expression, suggesting a potential role of miR-203 in preventing and treating keloids.