Using Human Primary Foreskin Fibroblasts to Study Cellular Damage and Mitochondrial Dysfunction

Nadalutti, Cristina A.; Wilson, S.H.

doi:10.1002/cptx.99

Cited by 4 publications

(3 citation statements)

References 29 publications

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“…HFF is a type of fibroblasts derived from foreskin tissues discarded after surgery. They have been used or proposed to use in several areas of research and medicine, including uses in cellular damage and mitochondrial dysfunction tests of toxins, feeder cells for stem cells, and wound healing [31][32][33][34]. Our data indicated that HUMSCs-MVs effectively reduced formation of hypertrophic scar tissues probably through inhibiting proliferation and invasion of fibroblasts, resolving inflammation and oxidative stress, suppressing Smad2/3 activation, and reducing angiogenesis.…”

Section: Introductionmentioning

confidence: 72%

Human Umbilical Mesenchymal Stem Cells-Derived Microvesicles Attenuate Formation of Hypertrophic Scar through Multiple Mechanisms

Qian

Zhu

Yang

et al. 2023

Stem Cells International

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Mesenchymal stem cells and the derived extracellular microvesicles are potential promising therapy for many disease conditions, including wound healing. Since current therapeutic approaches do not satisfactorily attenuate or ameliorate formation of hypertrophic scars, it is necessary to develop novel drugs to achieve better outcomes. In this study, we investigated the effects and the underlying mechanisms of human umbilical mesenchymal stem cells (HUMSCs)-derived microvesicles (HUMSCs-MVs) on hypertrophic scar formation using a rabbit ear model and a human foreskin fibroblasts (HFF) culture model. The results showed that HUMSCs-MVs reduced formation of hypertrophic scar tissues in the rabbit model based on appearance observation, and hematoxylin and eosin (H&E), Masson, and immunohistochemical stainings. HUMSCs-MVs inhibited invasion of HFF cells and decreased the levels of the α-SMA, N-WASP, and cortacin proteins. HUMSCs-MVs also inhibited cell proliferation of HFF cells. The MMP-1, MMP-3, and TIMP-3 mRNA levels were significantly increased, and the TIMP-4 mRNA level and the NF-kB p65/β-catenin protein levels were significantly decreased in HFF cells after HUMSCs-MVs treatment. The p-SMAD2/3 levels and the ratios of p-SMAD2/3/SMAD2/3 were significantly decreased in both the wound healing tissues and HFF cells after HUMSCs-MVs treatment. CD34 levels were significantly decreased in both wound healing scar tissues and HFF cells after HUMSCs-MVs treatment. The VEGF-A level was also significantly decreased in HFF cells after HUMSCs-MVs treatment. The magnitudes of changes in these markers by HUMSCs-MVs were mostly higher than those by dexamethasone. These results suggested that HUMSCs-MVs attenuated formation of hypertrophic scar during wound healing through inhibiting proliferation and invasion of fibrotic cells, inflammation and oxidative stress, Smad2/3 activation, and angiogenesis. HUMSCs-MVs is a potential promising drug to attenuate formation of hypertrophic scar during wound healing.

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Section: Introductionmentioning

confidence: 72%

Human Umbilical Mesenchymal Stem Cells-Derived Microvesicles Attenuate Formation of Hypertrophic Scar through Multiple Mechanisms

Qian

Zhu

Yang

et al. 2023

Stem Cells International

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“…Primary human dermal fibroblasts (isolated from healthy male neonatal foreskin tissues; Protocol number IRB 120/63 approved by the Institutional Review Board of the Faculty of Medicine, Chulalongkorn University) were grown in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, Life Technologies, Ltd., Paisley, Scotland, UK) supplemented with 10% fetal bovine serum (FBS) (Gibco, Life Technologies, Ltd.) in a humidified atmosphere at 37°C, 5% CO 2 . The primary human dermal fibroblasts (1 × 10 5 cells/well) in 12-well culture plates were incubated with either the Pseudomonas planktonic bacteria (1 × 10 4 CFU) or the sessile form, prepared by 1 × 10 4 CFU of bacteria on the polystyrene plates, at 37°C in the CO 2 incubator for 6 h. The protocol of primary fibroblasts isolation from a publication was followed [ 62 ]. Briefly, the foreskin tissues from healthy volunteers (IRB 120/63) were collected in sterile containers supplemented with DMEM and 10% FBS with 5 µg/mL of Plasmocin (the mycoplasma elimination reagent) (InvivoGen, San Diego, CA, USA).…”

Section: Methodsmentioning

confidence: 99%

Chlorhexidine Promotes Psl Expression in Pseudomonas aeruginosa That Enhances Cell Aggregation with Preserved Pathogenicity Demonstrates an Adaptation against Antiseptic

Singkham-in

Phuengmaung

Makjaroen

et al. 2022

IJMS

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Because Pseudomonas aeruginosa is frequently in contact with Chlorhexidine (a regular antiseptic), bacterial adaptations are possible. In comparison with the parent strain, the Chlorhexidine-adapted strain formed smaller colonies with metabolic downregulation (proteomic analysis) with the cross-resistance against colistin (an antibiotic for several antibiotic-resistant bacteria), partly through the modification of L-Ara4N in the lipopolysaccharide at the outer membrane. Chlorhexidine-adapted strain formed dense liquid–solid interface biofilms with enhanced cell aggregation partly due to the Chlorhexidine-induced overexpression of psl (exopolysaccharide-encoded gene) through the LadS/GacSA pathway (c-di-GMP-independence) in 12 h biofilms and maintained the aggregation with SiaD-mediated c-di-GMP dependence in 24 h biofilms as evaluated by polymerase chain reaction (PCR). The addition of Ca2+ in the Chlorhexidine-adapted strain facilitated several Psl-associated genes, indicating an impact of Ca2+ in Psl production. The activation by Chlorhexidine-treated sessile bacteria demonstrated a lower expression of IL-6 and IL-8 on fibroblasts and macrophages than the activation by the parent strain, indicating the less inflammatory reactions from Chlorhexidine-exposed bacteria. However, the 14-day severity of the wounds in mouse caused by Chlorhexidine-treated bacteria versus the parent strain was similar, as indicated by wound diameters and bacterial burdens. In conclusion, Chlorhexidine induced psl over-expression and colistin cross-resistance that might be clinically important.

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“…Foreskin cells have shown the ability to develop into a stable cell line of any of the three embryonic germ layers under in vitro conditions [3,4]. In addition, foreskin cells have been shown to be useful as a model for studying cellular damage and mitochondrial disfunction [5,6]. Moreover, the foreskin has been shown to be a good source for mesenchymal stromal cells that have several immunotherapeutic applications [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Attitude towards donation of the excised foreskin after circumcision surgery for research: A study from Madinah, Saudi Arabia

Khabour,

Mahallawi,

Ali

et al. 2023

PLoS ONE

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Studies have shown the possibility of using the part of the foreskin removed after circumcision in the field of scientific and therapeutic research. Donations of tissues and organs are always associated with ethical challenges posed by bioethicists and societies to ensure the appropriate use of these tissues/organs. The purpose of this study was to understand the attitudes and awareness of parents/guardians regarding donation of excised foreskin to research and medical use. The study was based on a questionnaire and included 133 parents/guardians who visited Uhud Children’s Hospital in Madinah, Saudi Arabia for newborn male circumcision. The results showed a high willingness (61.7%) to donate the extracted foreskin to research. The willingness to donate the extracted foreskin to research associated with undergraduate degree (P = 0.018), male sex (P = 0.011), high income (P = 0.029), and participation in previous research studies (P = 0.002). About 41.8% were convinced that written informed consent should be obtained before circumcision surgery, 38.1% (n = 51) were convinced that written informed consent should be taken after surgery, while the remaining 19.4% reported that the timing of written informed consent is unimportant. Finally, fear of excision of excess tissue (74.5%), lack of confidence in the research (68.6%), and potential for commercial use (64.7%) were the main barriers to unwillingness to donate the excised foreskin for research. In conclusion, a reasonable portion of Saudis agreed to donate their foreskin for research purposes. There is an urgent need to enhance awareness and attitudes towards tissue donation for research and therapeutic use.

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Using Human Primary Foreskin Fibroblasts to Study Cellular Damage and Mitochondrial Dysfunction

Abstract: Isolation and maintenance of human primary foreskin fibroblasts (FSK) Basic Protocol 2: Determination of cell viability by crystal violet staining Basic Protocol 3: Transmission electron microscopy to study cellular damage and mitochondrial dysfunction

Cited by 4 publications

References 29 publications

Human Umbilical Mesenchymal Stem Cells-Derived Microvesicles Attenuate Formation of Hypertrophic Scar through Multiple Mechanisms

Human Umbilical Mesenchymal Stem Cells-Derived Microvesicles Attenuate Formation of Hypertrophic Scar through Multiple Mechanisms

Chlorhexidine Promotes Psl Expression in Pseudomonas aeruginosa That Enhances Cell Aggregation with Preserved Pathogenicity Demonstrates an Adaptation against Antiseptic

Attitude towards donation of the excised foreskin after circumcision surgery for research: A study from Madinah, Saudi Arabia

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