Pseudomonas aeruginosa virulence proteins pseudolysin and protease IV impede cutaneous wound healing

AbstractThe intricate biological process of cutaneous wound healing is achieved through precise and highly programmed events. Dermal fibroblasts and keratinocytes play a significant role in the process of reepithelialization during wound healing. Pathogenic bacteria such as Pseudomonas aeruginosa (P. aeruginosa) may delay the proliferative phase of wound repair by secreting their proteins leading to delayed or impaired wound healing. We have analyzed three virulent strains of P. aeruginosa isolated from the wound environment which also differed in their ability to produce biofilms. Mass spectrometric analysis of differentially expressed secreted proteins by three virulent strains of P. aeruginosa revealed peptides from pseudolysin and protease IV expressed from lasB and prpL genes. Pseudolysin and protease IV recombinant proteins were tested for their ability to modulate wound healing in several cell types of wound microenvironment in in vitro and in vivo models. Both pseudolysin and protease IV inhibited migration and survival of fibroblasts, keratinocytes, and endothelial cells. In three dimensional spheroid endothelial models and matrigel assays these proteins impeded sprouting and tube formation. In a mouse model of excision wound, pseudolysin and protease IV treatment showed reduced collagen content, inhibited neovascularization and epithelialization, and delayed wound contraction. Furthermore, pseudolysin and protease IV treatment resulted in a significant increase in plasma IL-6 levels when compared to vehicle control and control, suggesting the induction of a state of prolonged inflammation. Taken together, our data indicate pseudolysin and protease IV secreted from biofilm producing and antibiotic resistant P. aeruginosa in wound microenvironment produce both local and systemic effects that is detrimental to the maintenance of tissue homeostasis. Hence, these proteins may serve as potential therapeutic targets toward better clinical management of wounds.

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Wound Healing and the Use of Medicinal Plants

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/2684108 ◽

2019 ◽

Vol 2019 ◽

pp. 1-30 ◽

Cited By ~ 17

Author(s):

Aleksandra Shedoeva ◽

David Leavesley ◽

Zee Upton ◽

Chen Fan

Keyword(s):

Wound Healing ◽

Natural Products ◽

Curcuma Longa ◽

Extended Period ◽

Centella Asiatica ◽

Dermal Fibroblasts ◽

Cutaneous Wound Healing ◽

Clinical Value ◽

Cutaneous Wound ◽

Traditional Medicines

Cutaneous wound healing is the process by which skin repairs itself. It is generally accepted that cutaneous wound healing can be divided into 4 phases: haemostasis, inflammation, proliferation, and remodelling. In humans, keratinocytes re-form a functional epidermis (reepithelialization) as rapidly as possible, closing the wound and reestablishing tissue homeostasis. Dermal fibroblasts migrate into the wound bed and proliferate, creating “granulation tissue” rich in extracellular matrix proteins and supporting the growth of new blood vessels. Ultimately, this is remodelled over an extended period, returning the injured tissue to a state similar to that before injury. Dysregulation in any phase of the wound healing cascade delays healing and may result in various skin pathologies, including nonhealing, or chronic ulceration. Indigenous and traditional medicines make extensive use of natural products and derivatives of natural products and provide more than half of all medicines consumed today throughout the world. Recognising the important role traditional medicine continues to play, we have undertaken an extensive survey of literature reporting the use of medical plants and plant-based products for cutaneous wounds. We describe the active ingredients, bioactivities, clinical uses, formulations, methods of preparation, and clinical value of 36 medical plant species. Several species stand out, including Centella asiatica, Curcuma longa, and Paeonia suffruticosa, which are popular wound healing products used by several cultures and ethnic groups. The popularity and evidence of continued use clearly indicates that there are still lessons to be learned from traditional practices. Hidden in the myriad of natural products and derivatives from natural products are undescribed reagents, unexplored combinations, and adjunct compounds that could have a place in the contemporary therapeutic inventory.

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Biomimetic RGD-engineered elastin-like extracellular matrix facilitates cutaneous wound healing in C57BL/6 mice by way of promoting the migration of epidermal keratinocytes and dermal fibroblasts

New Biotechnology ◽

10.1016/j.nbt.2012.08.424 ◽

2012 ◽

Vol 29 ◽

pp. S153 ◽

Cited By ~ 1

Author(s):

Jin-Kyu Park ◽

Seong-Kyoon Choi ◽

Kyu-Shik Jeong ◽

Kyeong-Min Lee ◽

Gwon-Soo Jung ◽

...

Keyword(s):

Extracellular Matrix ◽

Wound Healing ◽

Dermal Fibroblasts ◽

Epidermal Keratinocytes ◽

Cutaneous Wound Healing ◽

Cutaneous Wound

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Exosomes derived from autologous dermal fibroblasts promote diabetic cutaneous wound healing through the Akt/β-catenin pathway

Cell Cycle ◽

10.1080/15384101.2021.1894813 ◽

2021 ◽

Vol 20 (5-6) ◽

pp. 616-629

Author(s):

Xinye Han ◽

Peipei Wu ◽

Linli Li ◽

Hassan Mohamud Sahal ◽

Cheng Ji ◽

...

Keyword(s):

Wound Healing ◽

Dermal Fibroblasts ◽

Cutaneous Wound Healing ◽

Cutaneous Wound

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CCN4/WISP1 controls cutaneous wound healing by modulating proliferation, migration and ECM expression in dermal fibroblasts via α5β1 and TNFα

Matrix Biology ◽

10.1016/j.matbio.2018.01.004 ◽

2018 ◽

Vol 68-69 ◽

pp. 533-546 ◽

Cited By ~ 19

Author(s):

Mitsuaki Ono ◽

Asuka Masaki ◽

Azusa Maeda ◽

Tina M. Kilts ◽

Emilio S. Hara ◽

...

Keyword(s):

Wound Healing ◽

Dermal Fibroblasts ◽

Cutaneous Wound Healing ◽

Cutaneous Wound

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Antler stem cell-conditioned medium stimulates regenerative wound healing in rats

Stem Cell Research & Therapy ◽

10.1186/s13287-019-1457-9 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 6

Author(s):

Xiaoli Rong ◽

Wenhui Chu ◽

Haiying Zhang ◽

Yusu Wang ◽

Xiaoyan Qi ◽

...

Keyword(s):

Wound Healing ◽

Stem Cell ◽

Conditioned Medium ◽

Dermal Fibroblasts ◽

Cutaneous Wound Healing ◽

Cutaneous Wound ◽

Nih 3T3 ◽

Cell Conditioned Medium

Abstract Background When the deer antler is cast, it leaves a cutaneous wound that can achieve scarless healing due to the presence of antler stem cells (ASCs). This provides an opportunity to study regenerative wound healing. Methods In this study, we investigated the therapeutic effects and mechanism of antler stem cell-conditioned medium (ASC-CM) on cutaneous wound healing in rats. In vitro, we investigated the effects of the ASC-CM on proliferation of HUVEC and NIH-3T3 cell lines. In vivo, we evaluated the effects of ASC-CM on cutaneous wound healing using full-thickness skin punch-cut wounds in rats. Results The results showed that ASC-CM significantly stimulated proliferation of the HUVEC and NIH-3T3 cells in vitro. In vivo, completion of healing of the rat wounds treated with ASC-CM was on day 16 (± 3 days), 9 days (± 2 days) earlier than the control group (DMEM); the area of the wounds treated with ASC-CM was significantly smaller (p < 0.05) than the two control groups. Further molecular characterization showed that the ratios of Col3A1/Col1A2, TGF-β3/TGF-β1, MMP1/TIMP1, and MMP3/TIMP1 significantly increased (p < 0.01) in the healed tissue in the ASC-CM group. Conclusions In conclusion, ASC-CM effectively accelerated the wound closure rate and enhanced the quality of healing, which might be through transforming wound dermal fibroblasts into the fetal counterparts. Therefore, the ASC-CM may have potential to be developed as a novel cell-free therapeutic for scarless wound healing.

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Transient Senescence Induces Wound Healing Through SASP Factors: Therapeutic Potentials of Chick Early Amniotic Fluid (ceAF)

10.21203/rs.3.rs-80655/v1 ◽

2020 ◽

Author(s):

Ahmad Mashaal ◽

Yandi Sun ◽

Xueyun Zhang ◽

Zara Ahmad Khan ◽

Jin Qian ◽

...

Keyword(s):

Wound Healing ◽

Healing Process ◽

Medical Intervention ◽

Cutaneous Wound Healing ◽

Related Factors ◽

In Vivo Models ◽

Cutaneous Wound ◽

Higher Doses

Abstract Background Inflammatory, proliferative and remodelling phases constitute a cutaneous wound healing program. Therapeutic applications/medication are available; however, they commonly comprise fortified preservatives that might prolong the healing process. Chick early amniotic fluids (ceAF) contain native therapeutic factors with balanced chemokines, cytokines and growth-related factors; their origins in principle dictate no existence of harmful agents that would otherwise hamper embryo development. Instead, they possess a spectrum of molecules driving expeditious mitotic divisions and possibly exerting other functions. Methods Employing both in vitro and in vivo models, we examined ceAF’s therapeutic potentials in wound healing and found intriguing involvement of transient senescence, known to be intimately intermingled with Senescence Associated Secretory Phenotypes (SASP) that function in addition to or in conjunction with ceAF to facilitate wound healing. Results In our in vitro and in vivo cutaneous wound healing models, a low dose of ceAF exhibited the best efficacies; however, higher doses attenuated the wound healing, presumably by inducing p16 expression. Conclusion Our studies link an INK4/ARF locus-mediated signalling to cutaneous wound healing, implicate the therapeutic potentials of ceAF exerting functions likely by driving transient senescence and expediting cell proliferation, and conceptualize a homeostatic and/or balanced dosage strategy in medical intervention.

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Impaired Aquaporin 3 Expression in Reepithelialization of Cutaneous Wound Healing in the Diabetic Rat

Biological Research For Nursing ◽

10.1177/1099800412437032 ◽

2012 ◽

Vol 15 (3) ◽

pp. 347-355 ◽

Cited By ~ 10

Author(s):

Takashi Sugimoto ◽

Lijuan Huang ◽

Takeo Minematsu ◽

Yuko Yamamoto ◽

Mayumi Asada ◽

...

Keyword(s):

Wound Healing ◽

Messenger Rna ◽

Transmembrane Protein ◽

Diabetic Rat ◽

Cutaneous Wound Healing ◽

Necrosis Factor Alpha ◽

Impaired Wound Healing ◽

Aquaporin 3 ◽

Cutaneous Wound ◽

Tnf Α

Impaired cutaneous wound healing is a serious complication of diabetes mellitus (DM). Currently, little is known about reepithelialization in DM. However, recent studies identified aquaporin 3 (AQP3), a transmembrane protein that functions as a pore-like passive transporter, to be a key molecule in cutaneous epidermal wound healing. AQP3 expression is downregulated in response to tumor necrosis factor-alpha (TNF- α). Given that systemic TNF-α levels are functionally connected to impaired healing in diabetic mice and that both diabetic and Aqp3-deficient animals exhibit impaired reepithelialization, the authors hypothesized that impaired AQP3 expression might contribute to diabetes-impaired wound healing. In the present study, the authors examined AQP3 expression in the regenerating epidermis during cutaneous full thickness wound healing and in intact skin of a streptozotocin-induced diabetic rat model. Aqp3 messenger RNA expression levels were decreased in wounds of DM rats compared to controls. Immunohistochemical analysis showed an absence of AQP3 in the stratum spinosum of the regenerating epidermis in the DM group, whereas the stratum basale was positive for AQP3 in both groups. In summary, these findings suggest that there may be a relationship between impaired AQP3 expression and diabetes-delayed reepithelialization. Thus, future nursing studies should focus on this mechanism in diabetic wound healing.

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The Dishevelled-binding protein CXXC5 negatively regulates cutaneous wound healing

Journal of Experimental Medicine ◽

10.1084/jem.20141601 ◽

2015 ◽

Vol 212 (7) ◽

pp. 1061-1080 ◽

Cited By ~ 25

Author(s):

Soung-Hoon Lee ◽

Mi-Yeon Kim ◽

Hyun-Yi Kim ◽

Young-Mi Lee ◽

Heesu Kim ◽

...

Keyword(s):

Wound Healing ◽

Negative Feedback ◽

Critical Role ◽

Dermal Fibroblasts ◽

Epidermal Keratinocytes ◽

Binding Motif ◽

Cutaneous Wound Healing ◽

Collagen Production ◽

Cutaneous Wound

Wnt/β-catenin signaling plays important roles in cutaneous wound healing and dermal fibrosis. However, its regulatory mechanism has not been fully elucidated, and a commercially available wound-healing agent targeting this pathway is desirable but currently unavailable. We found that CXXC-type zinc finger protein 5 (CXXC5) serves as a negative feedback regulator of the Wnt/β-catenin pathway by interacting with the Dishevelled (Dvl) protein. In humans, CXXC5 protein levels were reduced in epidermal keratinocytes and dermal fibroblasts of acute wounds. A differential regulation of β-catenin, α-smooth muscle actin (α-SMA), and collagen I by overexpression and silencing of CXXC5 in vitro indicated a critical role for this factor in myofibroblast differentiation and collagen production. In addition, CXXC5−/− mice exhibited accelerated cutaneous wound healing, as well as enhanced keratin 14 and collagen synthesis. Protein transduction domain (PTD)–Dvl-binding motif (DBM), a competitor peptide blocking CXXC5-Dvl interactions, disrupted this negative feedback loop and activated β-catenin and collagen production in vitro. Co-treatment of skin wounds with PTD-DBM and valproic acid (VPA), a glycogen synthase kinase 3β (GSK3β) inhibitor which activates the Wnt/β-catenin pathway, synergistically accelerated cutaneous wound healing in mice. Together, these data suggest that CXXC5 would represent a potential target for future therapies aimed at improving wound healing.

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SPARC-thrombospondin-2-double-null Mice Exhibit Enhanced Cutaneous Wound Healing and Increased Fibrovascular Invasion of Subcutaneous Polyvinyl Alcohol Sponges

Journal of Histochemistry & Cytochemistry ◽

10.1369/jhc.4a6425.2005 ◽

2005 ◽

Vol 53 (5) ◽

pp. 571-581 ◽

Cited By ~ 24

Author(s):

Pauli A. Puolakkainen ◽

Amy D. Bradshaw ◽

Rolf A. Brekken ◽

May J. Reed ◽

Themis Kyriakides ◽

...

Keyword(s):

Wound Healing ◽

Wound Repair ◽

Dermal Fibroblasts ◽

Fine Tuning ◽

Cutaneous Wound Healing ◽

Collagen Gels ◽

Matricellular Proteins ◽

Cutaneous Wound ◽

Cutaneous Wounds

Secreted protein acidic and rich in cysteine (SPARC) and thrombospondin-2 (TSP-2) are structurally unrelated matricellular proteins that have important roles in cell- extracellular matrix (ECM) interactions and tissue repair. SPARC-null mice exhibit accelerated wound closure, and TSP-2-null mice show an overall enhancement in wound healing. To assess potential compensation of one protein for the other, we examined cutaneous wound healing and fibrovascular invasion of subcutaneous sponges in SPARC-TSP-2 (ST) double-null and wild-type (WT) mice. Epidermal closure of cutaneous wounds was found to occur significantly faster in ST-double-null mice, compared with WT animals: histological analysis of dermal wound repair revealed significantly more mature phases of healing at 1, 4, 7, 10, and 14 days after wounding, and electron microscopy showed disrupted ECM at 14 days in these mice. ST-double-null dermal fibroblasts displayed accelerated migration, relative to WT fibroblasts, in a wounding assay in vitro, as well as enhanced contraction of native collagen gels. Zymography indicated that fibroblasts from ST-double-null mice also produced higher levels of matrix metalloproteinase (MMP)-2. These data are consistent with the increased fibrovascular invasion of subcutaneous sponge implants seen in the double-null mice. The generally accelerated wound healing of ST-double-null mice reflects that described for the single-null animals. Importantly, the absence of both proteins results in elevated MMP-2 levels. SPARC and TSP-2 therefore perform similar functions in the regulation of cutaneous wound healing, but fine-tuning with respect to ECM production and remodeling could account for the enhanced response seen in ST-double-null mice.

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