Dynamics of Myosin II Filaments during Wound Repair in Dividing Cells

Wound repair of cell membranes is essential for cell survival. Myosin II contributes to wound pore closure by interacting with actin filaments in larger cells; however, its role in smaller cells is unclear. In this study, we observed wound repair in dividing cells for the first time. The cell membrane in the cleavage furrow, where myosin II localized, was wounded by laserporation. Upon wounding, actin transiently accumulated, and myosin II transiently disappeared from the wound site. Ca2+ influx from the external medium triggered both actin and myosin II dynamics. Inhibition of calmodulin reduced both actin and myosin II dynamics. The wound closure time in myosin II-null cells was the same as that in wild-type cells, suggesting that myosin II is not essential for wound repair. We also found that disassembly of myosin II filaments by phosphorylation did not contribute to their disappearance, indicating a novel mechanism for myosin II delocalization from the cortex. Furthermore, we observed that several furrow-localizing proteins such as GAPA, PakA, myosin heavy chain kinase C, PTEN, and dynamin disappeared upon wounding. Herein, we discuss the possible mechanisms of myosin dynamics during wound repair.

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Microtubule plus-end dynamics link wound repair to the innate immune response

10.1101/512632 ◽

2019 ◽

Author(s):

Clara Taffoni ◽

Shizue Omi ◽

Caroline Huber ◽

Sebastien Mailfert ◽

Matthieu Fallet ◽

...

Keyword(s):

Immune Response ◽

Innate Immune Response ◽

Wound Repair ◽

Wound Closure ◽

Immune Reaction ◽

Innate Immune ◽

First Line ◽

Physical Damage ◽

Wound Site ◽

C Elegans

AbstractAs a first line of defence against the environment, the epidermis protect animals from infection and physical damage. In C. elegans, wounding the epidermal epithelium triggers both an immune reaction and a repair response. Exactly how these are controlled, and the degree to which they are inter-connected remains unclear. To address these questions, we established a simple system for simultaneously inflicting precise laser wounds and imaging at high spatial and temporal resolution. We show that in C. elegans, wounding provokes a rapid sealing of the plasma membrane, involving reorganisation of phosphatidylinositol 4,5- bisphosphate domains. This is followed by a radial recruitment at the wound site of EBP-2/EB1, a protein that binds the plus ends of microtubules. EB1 recruitment is accompanied by a reorganisation of microtubules, required for the subsequent recruitment of actin and wound closure. It is also required for the directed trafficking towards the site of injury of the key signalling protein SNF-12. In the absence of SNF-12 recruitment, there is an abrogation of the immune response. Our results suggest that microtubule dynamics coordinate the cytoskeletal changes required for wound repair and the concomitant activation of the innate immune response.

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WD Repeat Domain of Dictyostelium Myosin Heavy Chain Kinase C Functions in both Substrate Targeting and Cellular Localization

Eukaryotic Cell ◽

10.1128/ec.00242-09 ◽

2009 ◽

Vol 9 (2) ◽

pp. 344-349

Author(s):

Atiya Franklin ◽

Linzi Hyatt ◽

Alyssa Chowdhury ◽

Paul A. Steimle

Keyword(s):

Myosin Heavy Chain ◽

Heavy Chain ◽

Cellular Localization ◽

Myosin Ii ◽

Content Type ◽

Kinase C ◽

Dictyostelium Myosin ◽

Repeat Domain ◽

Myosin Heavy Chain Kinase ◽

Wd Repeat

ABSTRACT Myosin II disassembly in Dictyostelium discoideum is regulated by three structurally related myosin heavy chain kinases (myosin II heavy chain kinase A [MHCK-A], -B, and -C). We show that the WD repeat domain of MHCK-C is unique in that it mediates both substrate targeting and subcellular localization, revealing a target for regulation that is distinct from those of the other MHCKs.

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Is a diluted seawater-based solution safe and effective on human nasal epithelium?

European Archives of Oto-Rhino-Laryngology ◽

10.1007/s00405-020-06527-1 ◽

2021 ◽

Author(s):

Song Huang ◽

Samuel Constant ◽

Barbara De Servi ◽

Marisa Meloni ◽

Amina Saaid ◽

...

Keyword(s):

Wound Repair ◽

Wound Closure ◽

Isotonic Saline ◽

Nasal Epithelium ◽

Mucin Secretion ◽

Nasal Irrigation ◽

Tissue Integrity ◽

Human Nasal Epithelium ◽

Pre Treatment

Abstract Purpose Nasal irrigation is an effective method for alleviating several nasal symptoms and regular seawater-based nasal irrigation is useful for maintaining nasal hygiene which is essential for appropriate functioning of the nose and for preventing airborne particles including some pollutants, pathogens, and allergens from moving further in the respiratory system. However, safety studies on seawater-based nasal irrigation are scarce. In this study, the safety and efficacy of a diluted isotonic seawater solution (Stérimar Nasal Hygiene, SNH) in maintaining nasal homeostasis were evaluated in vitro. Methods Safety was assessed by measuring tissue integrity via transepithelial electrical resistance (TEER). Efficacy was measured by mucociliary clearance (MCC), mucin secretion, and tissue re-epithelization (wound repair) assays. All assays were performed using a 3D reconstituted human nasal epithelium model. Results In SNH-treated tissues, TEER values were statistically significantly lower than the untreated tissues; however, the values were above the tissue integrity limit. SNH treatment significantly increased MCC (88 vs. 36 µm/s, p < 0.001) and mucin secretion (1717 vs. 1280 µg/ml, p < 0.001) as compared to untreated cultures. Faster wound closure profile was noted upon pre-SNH treatment as compared to classical isotonic saline solution pre-treatment (90.5 vs. 50.7% wound closure 22 h after wound generation). Conclusion SNH did not compromise the integrity of the nasal epithelium in vitro. Furthermore, SNH was effective for removal of foreign particles through MCC increase and for enhancing wound repair on nasal mucosa.

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Novel Myosin Heavy Chain Kinase Involved in Disassembly of Myosin II Filaments and Efficient Cleavage in MitoticDictyostelium Cells

Molecular Biology of the Cell ◽

10.1091/mbc.e02-04-0228 ◽

2002 ◽

Vol 13 (12) ◽

pp. 4333-4342 ◽

Cited By ~ 18

Author(s):

Akira Nagasaki ◽

Go Itoh ◽

Shigehiko Yumura ◽

Taro Q.P. Uyeda

Keyword(s):

Myosin Heavy Chain ◽

Heavy Chain ◽

Myosin Ii ◽

Kinase Domain ◽

Cleavage Furrow ◽

Wild Type ◽

Daughter Cells ◽

Cleavage Process ◽

Interphase Cells ◽

Myosin Heavy Chain Kinase

We have cloned a full-length cDNA encoding a novel myosin II heavy chain kinase (mhckC) from Dictyostelium. Like other members of the myosin heavy chain kinase family, themhckC gene product, MHCK C, has a kinase domain in its N-terminal half and six WD repeats in the C-terminal half. GFP-MHCK C fusion protein localized to the cortex of interphase cells, to the cleavage furrow of mitotic cells, and to the posterior of migrating cells. These distributions of GFP-MHCK C always corresponded with that of myosin II filaments and were not observed in myosin II-null cells, where GFP-MHCK C was diffusely distributed in the cytoplasm. Thus, localization of MHCK C seems to be myosin II-dependent. Cells lacking the mhckC gene exhibited excessive aggregation of myosin II filaments in the cleavage furrows and in the posteriors of the daughter cells once cleavage was complete. The cleavage process of these cells took longer than that of wild-type cells. Taken together, these findings suggest MHCK C drives the disassembly of myosin II filaments for efficient cytokinesis and recycling of myosin II that occurs during cytokinesis.

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Transforming growth factor-alpha enhances alveolar epithelial cell repair in a new in vitro model

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1994.267.6.l728 ◽

1994 ◽

Vol 267 (6) ◽

pp. L728-L738 ◽

Cited By ~ 37

Author(s):

F. Kheradmand ◽

H. G. Folkesson ◽

L. Shum ◽

R. Derynk ◽

R. Pytela ◽

...

Keyword(s):

Growth Factor ◽

Epithelial Cell ◽

Wound Repair ◽

Wound Closure ◽

Transforming Growth Factor ◽

Alveolar Epithelial Cell ◽

Alveolar Epithelial ◽

Factor Alpha ◽

Vitro Model

Alveolar epithelial type II cells are essential for regenerating an intact alveolar barrier after destruction of type I cells in vivo. The first objective of these experimental studies was to develop an in vitro model to quantify alveolar epithelial cell wound repair. The second objective was to investigate mechanisms of alveolar epithelial cell wound healing by studying the effects of serum and transforming growth factor-alpha (TGF-alpha) on wound closure. Primary cultures of rat alveolar type II cells were prepared by standard methods and grown to form confluent monolayers in 48 h. Then a wound was made by denuding an area (mean initial area of 2.1 +/- 0.6 mm2) of the monolayer. Re-epithelialization of the denuded area over time in the presence or absence of serum was measured using quantitative measurements from time-lapse video microscopy. The half time of wound healing was significantly enhanced in the presence of serum compared with serum-free conditions (2.4 +/- 0.2 vs. 17.4 +/- 0.8 h, P < 0.001). We then tested the hypothesis that TGF-alpha is an important growth factor for stimulating wound repair of alveolar epithelial cells. Exogenous addition of TGF-alpha in serum-free medium resulted in a significantly more rapid wound closure, and, furthermore, the addition of a monoclonal antibody to TGF-alpha in the presence of serum significantly decreased fourfold the rate of wound closure. Measurement of internuclear cell distance confirmed that both cell motility and cell spreading were responsible for closure of the wound. These data demonstrate that 1) the mechanisms of alveolar cell repair can be studied in vitro and that 2) TGF-alpha is a potent growth factor that enhances in vitro alveolar epithelial cell wound closure.

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Enhancement of wound closure in diabetic mice by ex vivo expanded cord blood CD34+ cells

Cellular & Molecular Biology Letters ◽

10.2478/s11658-013-0089-9 ◽

2013 ◽

Vol 18 (2) ◽

Cited By ~ 8

Author(s):

Kamonnaree Chotinantakul ◽

Chavaboon Dechsukhum ◽

Duangnapa Dejjuy ◽

Wilairat Leeanansaksiri

Keyword(s):

Cord Blood ◽

Wound Repair ◽

Wound Closure ◽

Ex Vivo ◽

Autologous Transplantation ◽

Diabetic Patients ◽

Isolated Cells ◽

Cytokine Cocktail ◽

Cd34 Cells ◽

Cord Blood Cd34

AbstractDiabetes can impair wound closure, which can give rise to major clinical problems. Most treatments for wound repair in diabetes remain ineffective. This study aimed to investigate the influence on wound closure of treatments using expanded human cord blood CD34+ cells (CB-CD34+ cells), freshly isolated CB-CD34+ cells and a cytokine cocktail. The test subjects were mice with streptozotocin-induced diabetes. Wounds treated with fresh CB-CD34+ cells showed more rapid repair than mice given the PBS control. Injection of expanded CB-CD34+ cells improved wound closure significantly, whereas the injection of the cytokine cocktail alone did not improve wound repair. The results also demonstrated a significant decrease in epithelial gaps and advanced re-epithelialization over the wound bed area after treatment with either expanded CB-CD34+ cells or freshly isolated cells compared with the control. In addition, treatments with both CB-CD34+ cells and the cytokine cocktail were shown to promote recruitment of CD31+-endothelial cells in the wounds. Both the CB-CD34+ cell population and the cytokine treatments also enhanced the recruitment of CD68-positive cells in the early stages (day 3) of treatment compared with PBS control, although the degree of this enhancement was found to decline in the later stages (day 9). These results demonstrated that expanded CB-CD34+ cells or freshly isolated CB-CD34+ cells could accelerate wound repair by increasing the recruitment of macrophages and capillaries and the reepithelialization over the wound bed area. Our data suggest an effective role in wound closure for both ex vivo expanded CB-CD34+ cells and freshly isolated cells, and these may serve as therapeutic options for wound treatment for diabetic patients. Wound closure acceleration by expanded CB-CD34+ cells also breaks the insufficient quantity obstacle of stem cells per unit of cord blood and other stem cell sources, which indicates a broader potential for autologous transplantation.

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The Difference in Incision Wound Closure Time Between 10% Povidone-iodine Solution and Chloramphenicol 2% Ointment in Swiss Webster Mice

Journal Of Medicine & Health ◽

10.28932/jmh.v2i6.2036 ◽

2020 ◽

Vol 2 (6) ◽

Author(s):

Selina Wissen ◽

Borman Sumaji ◽

Dian Lesmana

Keyword(s):

Wound Closure ◽

Povidone Iodine ◽

Iodine Solution ◽

Closure Time ◽

The Difference ◽

Incision Wound ◽

Povidone Iodine Solution

Luka adalah cedera fisik yang mengakibatkan rusaknya kulit. Penanganan luka diperlukan untuk mencegah terjadinya infeksi. Agen topikal untuk luka insisi ekstraoral yang umumnya tersedia di puskesmas dan klinik-klinik kesehatan umum maupun gigi yaitu solutiopovidone iodine 10% dan unguentum kloramfenikol 2%. Tujuan penelitian ini adalah untuk mengetahui apakah terdapat perbedaan waktu penutupan luka insisi yang diaplikasikan solutiopovidone iodine 10% dengan unguentum kloramfenikol 2% pada mencit Swiss Webster. Penelitian ini bersifat eksperimental laboratorik, menggunakan 30 ekor mencit Swiss Webster yang dibagi menjadi 2 kelompok. Kelompok I adalah luka insisi pada paha kanan mencit dan diaplikasikan solutiopovidone iodine 10%. Kelompok II adalah luka insisi pada paha kiri mencit dan diaplikasikan unguentum kloramfenikol 2%. Data yang diukur adalah rerata waktu penutupan luka insisi untuk kedua kelompok, kemudian dianalisis menggunakan uji non parametrik Mann-Whitney. Rerata waktu penutupan luka insisi yang diaplikasikan solutiopovidone iodine 10% adalah 5,07±0,691 hari, dan yang diaplikasikan unguentum kloramfenikol 2% adalah 5,03±0,765 hari. Simpulan penelitian adalah tidak terdapat perbedaan waktu penutupan luka insisi yang diaplikasikan solutio povidone iodine 10% dengan unguentum kloramfenikol 2% pada mencit Swiss Webster. Kata kunci: waktu penutupan luka, luka insisi, solutio povidone iodine 10%, unguentum kloramfenikol 2%

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Development of Haemoglobin by De-embryonated Chick Blastoderms Cultured in vitro and the Effect of Abnormal RNA upon its Synthesis

Development ◽

10.1242/dev.9.1.202 ◽

1961 ◽

Vol 9 (1) ◽

pp. 202-221

Author(s):

B. R. A. O'Brien

Keyword(s):

Protein Synthesis ◽

Developmental Stages ◽

Specific Protein ◽

Cytoplasmic Protein ◽

Whole Cells ◽

Restricted Group ◽

Dividing Cells ◽

First Time ◽

Structural Code

The embryo provides a sequence of developmental stages in which proteins both structural and enzymatic appear or become detectable for the first time in a restricted group of dividing cells. The cells or tissues can be maintained in vitro for a period that may precede and include the synthesis of a specific ‘cytoplasmic’ protein. In this way systems of protein synthesis within the cells of higher organisms can be studied during those stages in which current hypotheses suggest that some structural code is passed on from the DNA of the nucleus to the cytoplasm where the synthesis of the protein becomes maximal. Acellular preparations have contributed much to the elucidation of protein synthesis, but it is doubtful whether actual net synthesis has been obtained in systems less complex than the ‘protoplast’ developed by Spiegelman (1957). In order to study the synthesis of a specific protein it seems necessary at this stage to use whole cells.

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Management of Acute Wounds

10.2310/anes.2022 ◽

2019 ◽

Author(s):

Lee D. Faucher ◽

Angela L. Gibson

Keyword(s):

Wound Healing ◽

Wound Repair ◽

Wound Closure ◽

Wound Care ◽

Connective Tissue Diseases ◽

Skin Grafting ◽

Adjunctive Treatment ◽

Local Trauma ◽

Life Threatening ◽

Acute Wound

Acute wounds are the result of local trauma and may be associated with severe life-threatening injuries. All patients with acute wounds should be assessed for comorbidities such as malnutrition, diabetes, peripheral vascular disease, neuropathy, obesity, immune deficiency, autoimmune disorders, connective tissue diseases, coagulopathy, hepatic dysfunction, malignancy, smoking practices, medication use that could interfere with healing, and allergies. The authors address the key considerations in management of the acute wound, including anesthesia, location of wound repair (e.g. operating room or emergency department), hemostasis, irrigation, débridement, closure materials, timing and methods of closure, adjunctive treatment (e.g. tetanus and rabies prophylaxis, antibiotics, and nutritional supplementation), appropriate closure methods for specific wound types, dressings, postoperative wound care, and potential disturbances of wound healing. This review contains 11 figures, 31 tables, and 92 references. Keywords: wound, wound infection, burns, suture, staple, wound closure, wound healing, dehiscence, skin grafting

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A symmetric anchor designed barbed suture versus conventional interrupted sutures in total knee arthroplasty: A multicenter, randomized controlled trial

Journal of Orthopaedic Surgery ◽

10.1177/2309499020965681 ◽

2020 ◽

Vol 28 (3) ◽

pp. 230949902096568

Author(s):

Wei Wang ◽

Shigui Yan ◽

Feng Liu ◽

Wei Chai ◽

Jianlin Zuo ◽

...

Keyword(s):

Total Knee Arthroplasty ◽

Randomized Controlled Trial ◽

Knee Arthroplasty ◽

Wound Closure ◽

Controlled Trial ◽

Barbed Suture ◽

Surgical Incision ◽

Randomized Controlled ◽

Closure Time ◽

Total Knee

Objective: This randomized controlled study was designed to compare the wound closure efficacy and safety of barbed suture in comparison to the conventional interrupted suture for total knee arthroplasty (TKA). Methods: This multicenter, single-blind, randomized controlled trial enrolled 184 patients who underwent elective TKA between June 2017 and April 2018. The subjects were randomized between two groups. Surgical incision closure time was considered as the primary end point. Results: A total of 184 patients participated in this randomized controlled trial; 91 patients had wound closure that involved barbed suture and 93 patients underwent conventional treatment—that is interrupted suturing with nonbarbed sutures. The surgical incision closure time was shorter ( p < 0.0001) in the barbed suture group compared with the control group (15.5 ± 4.88 vs. 20.9 ± 6.30 min). However, both groups were found to be equal in terms of the rate of postoperative complications. Conclusion: Usage of the symmetric anchor designed barbed suture is safe, efficacious, and demonstrates a decrease in surgical incision closure time in patients undergoing TKA compared to interrupted closure using conventional sutures. Future studies are warranted to demonstrate clinical and economic benefits of barbed sutures.

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