Evaluation of In Vitro Wound-Healing Potential, Antioxidant Capacity, and Antimicrobial Activity of Stellaria media (L.) Vill

The healing of skin wounds remains an important concern in medicine, especially in chronic wounds caused by various diseases such as diabetes. Using herbs or herbal products to heal skin wounds is a therapeutic challenge for traditional medicine. In this context, the main aim of our work was to highlight the in vitro healing potential of Stellaria media (L.) Vill. (SM) extract using the scratch assay on normal human dermal fibroblasts (NHDF). The ability to stimulate cell migration and proliferation under the influence of different concentrations of SM extract (range between 12.5 and 200 µg/mL) was determined compared to the control (untreated in vitro-simulated wound) and positive control (allantoin 50 µg/mL). Our results showed that the concentration of 100 µg/mL SM extract applied on the simulated wound recorded the strongest and fastest (24 h) migration (with wound closure) and proliferation of NHDF compared with the control. In addition, the SM extract was characterized in terms of bioactive compounds (total phenols and flavonoids content), antioxidant capacity (FRAP (The Ferric-Reducing Antioxidant Power) assay and electrochemical method), and antimicrobial activity. The results show that the SM extract contains a considerable amount of polyphenols (17.19 ± 1.32 mg GAE/g dw and 7.28 ± 1.18 mg QE/g dw for total phenol and flavonoid content, respectively) with antioxidant capacity. Antimicrobial activity against Gram-positive bacteria (S. aureus) is higher than E. coli at a dose of 15 µg/mL. This study showed that Stellaria media is a source of polyphenols compounds with antioxidant capacity, and for the first time, its wound healing potential was emphasized.

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Dermal fibroblasts cultured from donors with type 2 diabetes mellitus retain an epigenetic memory associated with poor wound healing responses

Scientific Reports ◽

10.1038/s41598-020-80072-z ◽

2021 ◽

Vol 11 (1) ◽

Cited By ~ 1

Author(s):

Aaiad H. A. Al-Rikabi ◽

Desmond J. Tobin ◽

Kirsten Riches-Suman ◽

M. Julie Thornton

Keyword(s):

Diabetes Mellitus ◽

Type 2 Diabetes ◽

Wound Healing ◽

Type 2 Diabetes Mellitus ◽

Chronic Wounds ◽

Dermal Fibroblasts ◽

Necrosis Factor Alpha ◽

Tnf Α

AbstractThe prevalence of Type 2 diabetes mellitus (T2DM) is escalating globally. Patients suffer from multiple complications including the development of chronic wounds that can lead to amputation. These wounds are characterised by an inflammatory environment including elevated tumour necrosis factor alpha (TNF-α). Dermal fibroblasts (DF) are critical for effective wound healing, so we sought to establish whether there were any differences in DF cultured from T2DM donors or those without diabetes (ND-DF). ND- and T2DM-DF when cultured similarly in vitro secreted comparable concentrations of TNF-α. Functionally, pre-treatment with TNF-α reduced the proliferation of ND-DF and transiently altered ND-DF morphology; however, T2DM-DF were resistant to these TNF-α induced changes. In contrast, TNF-α inhibited ND- and T2DM-DF migration and matrix metalloprotease expression to the same degree, although T2DM-DF expressed significantly higher levels of tissue inhibitor of metalloproteases (TIMP)-2. Finally, TNF-α significantly increased the secretion of pro-inflammatory cytokines (including CCL2, CXCL1 and SERPINE1) in ND-DF, whilst this effect in T2DM-DF was blunted, presumably due to the tendency to higher baseline pro-inflammatory cytokine expression observed in this cell type. Collectively, these data demonstrate that T2DM-DF exhibit a selective loss of responsiveness to TNF-α, particularly regarding proliferative and secretory functions. This highlights important phenotypic changes in T2DM-DF that may explain the susceptibility to chronic wounds in these patients.

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Biomimetic In Vitro Model of Cell Infiltration into Skin Scaffolds for Pre-Screening and Testing of Biomaterial-Based Therapies

Cells ◽

10.3390/cells8080917 ◽

2019 ◽

Vol 8 (8) ◽

pp. 917

Author(s):

Rafael Ballesteros-Cillero ◽

Evan Davison-Kotler ◽

Nupur Kohli ◽

William S. Marshall ◽

Elena García-Gareta

Keyword(s):

Wound Healing ◽

In Vitro Model ◽

Chronic Wounds ◽

Healing Process ◽

Dermal Fibroblasts ◽

Surrounding Tissue ◽

Cell Infiltration ◽

Skin Substitutes ◽

Vitro Model

Due to great clinical need, research where different biomaterials are tested as 3D scaffolds for skin tissue engineering has increased. In vitro studies use a cell suspension that is simply pipetted onto the material and cultured until the cells migrate and proliferate within the 3D scaffold, which does not mimic the in vivo reality. Our aim was to engineer a novel biomimetic in vitro model that mimics the natural cell infiltration process occurring in wound healing, thus offering a realistic approach when pre-screening and testing new skin substitutes. Our model consists of porous membrane cell culture inserts coated with gelatin and seeded with human dermal fibroblasts, inside which two different commercially available dermal substitutes were placed. Several features relevant to the wound healing process (matrix contraction, cell infiltration and proliferation, integration of the biomaterial with the surrounding tissue, and secretion of exogenous cytokines and growth factors) were evaluated. Our results showed that cells spontaneously infiltrate the materials and that our engineered model is able to induce and detect subtle differences between different biomaterials. The model allows for room for improvements or “adds-on” and miniaturization and can contribute to the development of functional and efficient skin substitutes for burns and chronic wounds.

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Effect of Bioactive Phytochemicals from Phlomis viscosa Poiret on Wound Healing

Plants ◽

10.3390/plants8120609 ◽

2019 ◽

Vol 8 (12) ◽

pp. 609 ◽

Cited By ~ 3

Author(s):

Ludmila Yarmolinsky ◽

Arie Budovsky ◽

Leonid Yarmolinsky ◽

Boris Khalfin ◽

Vladimir Glukhman ◽

...

Keyword(s):

Wound Healing ◽

Beneficial Effect ◽

Chronic Wounds ◽

Dermal Fibroblasts ◽

Human Dermal Fibroblasts ◽

In Vivo Models ◽

Bioactive Phytochemicals ◽

Pro Inflammatory Cytokines

Phlomis viscosa Poiret is an evergreen shrub growing in Israel, Turkey, Lebanon, and Syria with acknowledged pro-wound healing (WH) properties. In this study, we evaluated the pro-WH potential of selected compounds found in this plant. Among the pro-WH compounds (identified by us) was a combination of three chemicals—diosmin, 1-octen-3-ol, and himachala-2,4-diene which enhanced WH significantly both in in vitro and in vivo models. The determined phytochemicals combination could be used for the treatment of chronic wounds. The effect of the extracts, diosmin, 1-octen-3-ol on the secretion of pro-inflammatory cytokines, IL-6 (A) and IL-8 (B) by human dermal fibroblasts was significant (p < 0.001). In addition, the beneficial effect of extracts of P. viscosa and its phytochemicals on WH was evidenced by inhibiting the growth of several WH delaying microorganisms.

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Fibrin-Modified Cellulose as a Promising Dressing for Accelerated Wound Healing

Materials ◽

10.3390/ma11112314 ◽

2018 ◽

Vol 11 (11) ◽

pp. 2314 ◽

Cited By ~ 6

Author(s):

Marketa Bacakova ◽

Julia Pajorova ◽

Tomas Sopuch ◽

Lucie Bacakova

Keyword(s):

Cell Proliferation ◽

Wound Healing ◽

Wound Dressing ◽

Chronic Wounds ◽

Cell Attachment ◽

Cellulose Fibers ◽

Dermal Fibroblasts ◽

Hydroxyl Groups ◽

Structural Morphology

Dermal injuries and chronic wounds usually regenerate with scar formation. Successful treatment without scarring might be achieved by pre-seeding a wound dressing with cells. We aimed to prepare a wound dressing fabricated from sodium carboxymethylcellulose (Hcel® NaT), combined with fibrin and seeded with dermal fibroblasts in vitro. We fabricated the Hcel® NaT in a porous and homogeneous form (P form and H form, respectively) differing in structural morphology and in the degree of substitution of hydroxyl groups. Each form of Hcel® NaT was functionalized with two morphologically different fibrin structures to improve cell adhesion and proliferation, estimated by an MTS assay. Fibrin functionalization of the Hcel® NaT strongly enhanced colonization of the material with human dermal fibroblasts. Moreover, the type of fibrin structures influenced the ability of the cells to adhere to the material and proliferate on it. The fibrin mesh filling the void spaces between cellulose fibers better supported cell attachment and subsequent proliferation than the fibrin coating, which only enwrapped individual cellulose fibers. On the fibrin mesh, the cell proliferation activity on day 3 was higher on the H form than on the P form of Hcel® NaT, while on the fibrin coating, the cell proliferation on day 7 was higher on the P form. The Hcel® NaT wound dressing functionalized with fibrin, especially when in the form of a mesh, can accelerate wound healing by supporting fibroblast adhesion and proliferation.

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Evaluation of the In Vitro Wound-Healing Activity of Calabrian Honeys

Antioxidants ◽

10.3390/antiox8020036 ◽

2019 ◽

Vol 8 (2) ◽

pp. 36 ◽

Cited By ~ 10

Author(s):

Paolo Governa ◽

Gabriele Carullo ◽

Marco Biagi ◽

Vittoria Rago ◽

Francesca Aiello

Keyword(s):

Wound Healing ◽

Chronic Wounds ◽

Biological Activities ◽

Folk Medicine ◽

Gas Chromatography Mass Spectrometry ◽

Skin Wounds ◽

Therapeutic Tools ◽

The Many ◽

Wound Healing Activity

The healing of skin wounds and particularly chronic wounds, such as diabetic foot ulcers, is still a clinical emergency. Despite the many therapeutic tools that are available so far, none seems to be really effective and safe. In this context, we highlighted the renewed wound healing activity of honey, a viscous aromatic and sweet food, by way of in vitro wound-healing assays, using the HaCaT cell line. Specifically, we investigated five monofloral or multifloral honeys from different Calabrian provinces using them as such or extracted (by Amberlite® or n-hexane and ethyl acetate). The chemical composition of honeys was ascertained by 1H NMR spectroscopy and by the gas chromatography/mass spectrometry (GC/MS) method for volatile organic compounds (VOCs). Amongst the five tested honeys, BL1 and BL5 honeys showed the most promising healing properties. Pinocembrin, which was revealed in BL1 (multifloral) and BL5 (orange) honey samples, is a flavanol that is already known to possess interesting biological activities, including healing. This study aims to investigate how a traditional food such as honey, which is appreciated for its nutritional value and used in folk medicine, can be enhanced as an effective modern remedial to promote a multifaceted and safe healing activity for all skin wounds.

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Development of Antibacterial Protective Coatings Active against MSSA and MRSA on Biodegradable Polymers

Polymers ◽

10.3390/polym13040659 ◽

2021 ◽

Vol 13 (4) ◽

pp. 659

Author(s):

Iva Rezić ◽

Mislav Majdak ◽

Vanja Ljoljić Bilić ◽

Ivan Pokrovac ◽

Lela Martinaga ◽

...

Keyword(s):

Antimicrobial Activity ◽

Biodegradable Polymers ◽

Chronic Wounds ◽

Protective Coatings ◽

Metal Oxide Nanoparticles ◽

Colloidal Solutions ◽

Fused Deposition ◽

Antimicrobial Nanoparticles ◽

Mrsa Strains

In this work the in vitro antimicrobial activity of colloidal solutions of nine different commercially available nanoparticles were investigated against Staphylococcus aureus strains, both methicillin-sensitive (MSSA) and methicillin-resistant (MRSA). Research covered antimicrobial investigation of different metal and metal-oxide nanoparticles, including Ag 10 nm, Ag 40 nm, Al2O3 100 nm, Au 20 nm, Pt 4 nm, TiO2 100 nm, Y2O3 100 nm, ZnO 100 nm and ZrO2 100 nm nanoparticles. Such materials were foreseen to be applied as coatings on 3D-printed biodegradable polymers: i.e., catheters, disposable materials, hospital bedding items, disposable antimicrobial linings and bandages for chronic wounds. Therefore, the antimicrobial activity of the nanoparticles was determined by agar well diffusion assays and serial microdilution broth assays. In addition, the chromatographic characterization of elements present in trace amounts was performed as a method for tracing the nanoparticles. Moreover, the potential of preparing the rough surface of biodegradable polymers for coating with antimicrobial nanoparticles was tested by 3D-printing fused deposition methodology. The in vitro results have shown that particular nanoparticles provided powerful antimicrobial effects against MSSA and MRSA strains, and can be easily applied on different biopolymers.

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Systematic Review: Adipose-Derived Mesenchymal Stem Cells, Platelet-Rich Plasma and Biomaterials as New Regenerative Strategies in Chronic Skin Wounds and Soft Tissue Defects

International Journal of Molecular Sciences ◽

10.3390/ijms22041538 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1538 ◽

Cited By ~ 2

Author(s):

Pietro Gentile ◽

Simone Garcovich

Keyword(s):

Systematic Review ◽

Stem Cells ◽

Wound Healing ◽

Soft Tissue ◽

Chronic Wounds ◽

Platelet Rich Plasma ◽

Skin Wounds ◽

Soft Tissue Defects ◽

Chronic Skin ◽

Tissue Defects

The number of clinical trials evaluating adipose-derived mesenchymal stem cells (AD-MSCs), platelet-rich plasma (PRP), and biomaterials efficacy in regenerative plastic surgery has exponentially increased during the last ten years. AD-MSCs are easily accessible from various fat depots and show intrinsic plasticity in giving rise to cell types involved in wound healing and angiogenesis. AD-MSCs have been used in the treatment of soft tissue defects and chronic wounds, employed in conjunction with a fat grafting technique or with dermal substitute scaffolds and platelet-rich plasma. In this systematic review, an overview of the current knowledge on this topic has been provided, based on existing studies and the authors’ experience. A multistep search of the PubMed, MEDLINE, Embase, PreMEDLINE, Ebase, CINAHL, PsycINFO, Clinicaltrials.gov, Scopus database, and Cochrane databases has been performed to identify papers on AD-MSCs, PRP, and biomaterials used in soft tissue defects and chronic wounds. Of the 2136 articles initially identified, 422 articles focusing on regenerative strategies in wound healing were selected and, consequently, only 278 articles apparently related to AD-MSC, PRP, and biomaterials were initially assessed for eligibility. Of these, 85 articles were excluded as pre-clinical, experimental, and in vitro studies. For the above-mentioned reasons, 193 articles were selected; of this amount, 121 letters, expert opinions, commentary, and editorials were removed. The remaining 72 articles, strictly regarding the use of AD-MSCs, PRP, and biomaterials in chronic skin wounds and soft tissue defects, were analyzed. The studies included had to match predetermined criteria according to the patients, intervention, comparator, outcomes, and study design (PICOS) approach. The information analyzed highlights the safety and efficacy of AD-MSCs, PRP, and biomaterials on soft tissue defects and chronic wounds, without major side effects.

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Chronic senescent human mesenchymal stem cells as possible contributor to the wound healing disorder after exposure to the alkylating agent sulfur mustard

Archives of Toxicology ◽

10.1007/s00204-020-02946-5 ◽

2021 ◽

Vol 95 (2) ◽

pp. 727-747

Author(s):

Simone Rothmiller ◽

Niklas Jäger ◽

Nicole Meier ◽

Thimo Meyer ◽

Adrian Neu ◽

...

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Mesenchymal Stem Cells ◽

Alkylating Agent ◽

Chronic Wounds ◽

Sulfur Mustard ◽

Morphological Changes ◽

Human Mesenchymal Stem Cells ◽

Age Related

AbstractWound healing is a complex process, and disturbance of even a single mechanism can result in chronic ulcers developing after exposure to the alkylating agent sulfur mustard (SM). A possible contributor may be SM-induced chronic senescent mesenchymal stem cells (MSCs), unable to fulfil their regenerative role, by persisting over long time periods and creating a proinflammatory microenvironment. Here we show that senescence induction in human bone marrow derived MSCs was time- and concentration-dependent, and chronic senescence could be verified 3 weeks after exposure to between 10 and 40 µM SM. Morphological changes, reduced clonogenic and migration potential, longer scratch closure times, differences in senescence, motility and DNA damage response associated genes as well as increased levels of proinflammatory cytokines were revealed. Selective removal of these cells by senolytic drugs, in which ABT-263 showed initial potential in vitro, opens the possibility for an innovative treatment strategy for chronic wounds, but also tumors and age-related diseases.

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The Designer Antimicrobial Peptide A-hBD-2 Facilitates Skin Wound Healing by Stimulating Keratinocyte Migration and Proliferation

Cellular Physiology and Biochemistry ◽

10.1159/000495320 ◽

2018 ◽

Vol 51 (2) ◽

pp. 647-663 ◽

Cited By ~ 2

Author(s):

Bobin Mi ◽

Jing Liu ◽

Yi Liu ◽

Liangcong Hu ◽

Yukun Liu ◽

...

Keyword(s):

Antimicrobial Activity ◽

Wound Healing ◽

Antimicrobial Peptide ◽

Structural Stability ◽

Cell Counting ◽

Sprague Dawley ◽

Skin Wound Healing ◽

High Sodium

Background/Aims: Antimicrobial peptides are effective promoters of wound healing but are susceptible to degradation. In this study, we replaced the GIGDP unit on the N-terminal of the endogenous human antimicrobial peptide hBD-2 with APKAM to produce A-hBD-2 and analyzed the effect on wound healing both in vitro and in vivo. Methods: The effects of A-hBD-2 and hBD-2 on cytotoxicity and proliferation in keratinocytes were assessed by Cell Counting Kit-8 assay. The structural stability and antimicrobial activity of hBD-2 and A-hBD-2 were evaluated against Staphylococcus aureus. RNA and proteins levels were evaluated by real-time PCR and western blotting, respectively. Cell migration was evaluated using a transwell assay. Cell cycle analysis was performed by flow cytometry. Wound healing was assessed in Sprague-Dawley rats. Epidermal thickness was evaluated by hematoxylin and eosin staining. Results: We found that hBD-2 exhibited cytotoxicity at high concentrations and decreased the structural stability in the presence of high sodium chloride concentrations. A-hBD-2 exhibited increased structural stability and antimicrobial activity, and had lower cytotoxicity in keratinocytes. A-hBD-2 increased the migration and proliferation of keratinocytes via phosphorylation of EGFR and STAT3 and suppressed terminal differentiation of keratinocytes. We also found that A-hBD-2 elicited mobilization of intracellular Ca2+ and stimulated keratinocytes to produce pro- and anti-inflammatory cytokines and chemokines via phospholipase C activation. Furthermore, A-hBD-2 promoted wound healing in vivo. Conclusion: Our data suggest that A-hBD-2 may be a promising candidate therapy for wound healing.

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Influence of Antihypertensive and Antidiabetic Drugs on Wound Healing – Essentials of a Systematic In-Vitro Study

Dermatology and Dermatitis ◽

10.31579/2578-8949/050 ◽

2019 ◽

Vol 4 (1) ◽

pp. 01-04

Author(s):

Ewa K. Stuermer

Keyword(s):

Wound Healing ◽

Life Style ◽

Chronic Wounds ◽

In Vitro Study ◽

Antidiabetic Drugs ◽

Vitro Study ◽

Lower Limbs ◽

Nerve Lesions

Due to the increasing prevalence of so called "life style diseases", such as diabetes, obesity or hypertension, the number of associated vascular and nerve lesions increases. In the lower limbs in particular, bagatelle trauma causes wounds that lead to wound healing disorders and chronic wounds

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