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Modeling the roles of protein kinase Cβ and η in single-cell wound repair.
Holmes WR, Liao L, Bement W, Edelstein-Keshet L
(2015) Mol Biol Cell 26: 4100-8
MeSH Terms: Actin Cytoskeleton, Actins, Animals, Models, Biological, Oocytes, Protein Kinase C, Protein Kinase C beta, Single-Cell Analysis, Wound Healing, Xenopus laevis, rho GTP-Binding Proteins
Show Abstract · Added February 26, 2016
Wounded cells such as Xenopus oocytes respond to damage by assembly and closure of an array of actin filaments and myosin-2 controlled by Rho GTPases, including Rho and Cdc42. Rho and Cdc42 are patterned around wounds in a characteristic manner, with active Rho concentrating in a ring-like zone inside a larger, ring-like zone of active Cdc42. How this patterning is achieved is unknown, but Rho and Cdc42 at wounds are subject to regulation by other proteins, including the protein kinases C. Specifically, Cdc42 and Rho activity are enhanced by PKCβ and inhibited by PKCη. We adapt a mathematical model of Simon and coworkers to probe the possible roles of these kinases. We show that PKCβ likely affects the magnitude of positive Rho-Abr feedback, whereas PKCη acts on Cdc42 inactivation. The model explains both qualitative and some overall quantitative features of PKC-Rho GTPase regulation. It also accounts for the previous, peculiar observation that ∼ 20% of cells overexpressing PKCη display zone inversions--that is, displacement of active Rho to the outside of the active Cdc42.
© 2015 Holmes, Liao, et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).
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11 MeSH Terms
Inhibition of Wnt/β-catenin pathway promotes regenerative repair of cutaneous and cartilage injury.
Bastakoty D, Saraswati S, Cates J, Lee E, Nanney LB, Young PP
(2015) FASEB J 29: 4881-92
MeSH Terms: Animals, Cartilage, Hair Follicle, Mice, Mice, Inbred C57BL, Regeneration, Skin, Wnt Proteins, Wound Healing, beta Catenin
Show Abstract · Added November 2, 2015
Wound healing in mammals is a fibrotic process. The mechanisms driving fibrotic (as opposed to regenerative) repair are poorly understood. Herein we report that therapeutic Wnt inhibition with topical application of small-molecule Wnt inhibitors can reduce fibrosis and promote regenerative cutaneous wound repair. In the naturally stented model of ear punch injury, we found that Wnt/β-catenin pathway is activated most notably in the dermis of the wound bed early (d 2) after injury and subsides to baseline levels by d10. Topical application of either of 2 mechanistically distinct small-molecule Wnt pathway inhibitors (a tankyrase inhibitor, XAV-939, and the U.S. Food and Drug Administration-approved casein kinase activator, pyrvinium) in C57Bl/6J mice resulted in significantly increased rates of wound closure (72.3 ± 14.7% with XAV-939; and 52.1 ± 20.9% with pyrvinium) compared with contralateral controls (38.1 ± 23.0 and 40.4.± 16.7%, respectively). Histologically, Wnt inhibition reduced fibrosis as measured by α-smooth muscle actin positive myofibroblasts and collagen type I α1 synthesis. Wnt inhibition also restored skin architecture including adnexal structures in ear wounds and dermal-epidermal junction with rete pegs in excisional wounds. Additionally, in ear punch injury Wnt inhibitor treatment enabled regeneration of auricular cartilage. Our study shows that pharmacologic Wnt inhibition holds therapeutic utility for regenerative repair of cutaneous wounds.
© FASEB.
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10 MeSH Terms
Myocardial Infarction Activates CCR2(+) Hematopoietic Stem and Progenitor Cells.
Dutta P, Sager HB, Stengel KR, Naxerova K, Courties G, Saez B, Silberstein L, Heidt T, Sebas M, Sun Y, Wojtkiewicz G, Feruglio PF, King K, Baker JN, van der Laan AM, Borodovsky A, Fitzgerald K, Hulsmans M, Hoyer F, Iwamoto Y, Vinegoni C, Brown D, Di Carli M, Libby P, Hiebert SW, Scadden DT, Swirski FK, Weissleder R, Nahrendorf M
(2015) Cell Stem Cell 16: 477-87
MeSH Terms: Animals, Cell Movement, Cells, Cultured, Hematopoietic Stem Cells, Macrophages, Mice, Inbred C57BL, Mice, Knockout, Mice, Mutant Strains, Models, Animal, Monocytes, Myeloid Cells, Myelopoiesis, Myocardial Infarction, Nuclear Proteins, RNA, Small Interfering, Receptors, CCR2, Transcription Factors, Wound Healing
Show Abstract · Added September 28, 2015
Following myocardial infarction (MI), myeloid cells derived from the hematopoietic system drive a sharp increase in systemic leukocyte levels that correlates closely with mortality. The origin of these myeloid cells, and the response of hematopoietic stem and progenitor cells (HSPCs) to MI, however, is unclear. Here, we identify a CCR2(+)CD150(+)CD48(-) LSK hematopoietic subset as the most upstream contributor to emergency myelopoiesis after ischemic organ injury. This subset has 4-fold higher proliferation rates than CCR2(-)CD150(+)CD48(-) LSK cells, displays a myeloid differentiation bias, and dominates the migratory HSPC population. We further demonstrate that the myeloid translocation gene 16 (Mtg16) regulates CCR2(+) HSPC emergence. Mtg16(-/-) mice have decreased levels of systemic monocytes and infarct-associated macrophages and display compromised tissue healing and post-MI heart failure. Together, these data provide insights into regulation of emergency hematopoiesis after ischemic injury and identify potential therapeutic targets to modulate leukocyte output after MI.
Copyright © 2015 Elsevier Inc. All rights reserved.
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18 MeSH Terms
MiRNA inhibition in tissue engineering and regenerative medicine.
Beavers KR, Nelson CE, Duvall CL
(2015) Adv Drug Deliv Rev 88: 123-37
MeSH Terms: Bone and Bones, Cicatrix, Genetic Vectors, Humans, Inflammation, Kidney, Liver, MicroRNAs, Muscle, Skeletal, Myocardium, Neovascularization, Pathologic, RNA, Messenger, Regeneration, Regenerative Medicine, Tissue Engineering, Tissue Scaffolds, Wound Healing
Show Abstract · Added March 14, 2018
MicroRNAs (miRNAs) are noncoding RNAs that provide an endogenous negative feedback mechanism for translation of messenger RNA (mRNA) into protein. Single miRNAs can regulate hundreds of mRNAs, enabling miRNAs to orchestrate robust biological responses by simultaneously impacting multiple gene networks. MiRNAs can act as master regulators of normal and pathological tissue development, homeostasis, and repair, which has motivated expanding efforts toward the development of technologies for therapeutically modulating miRNA activity for regenerative medicine and tissue engineering applications. This review highlights the tools currently available for miRNA inhibition and their recent therapeutic applications for improving tissue repair.
Copyright © 2014 Elsevier B.V. All rights reserved.
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17 MeSH Terms
Imaging mass spectrometry for assessing cutaneous wound healing: analysis of pressure ulcers.
Taverna D, Pollins AC, Sindona G, Caprioli RM, Nanney LB
(2015) J Proteome Res 14: 986-96
MeSH Terms: Adolescent, Adult, Aged, Female, Humans, Male, Mass Spectrometry, Middle Aged, Molecular Imaging, Pressure Ulcer, Proteome, Proteomics, S100 Proteins, Wound Healing, Young Adult
Show Abstract · Added February 12, 2015
Imaging mass spectrometry (IMS) was employed for the analysis of frozen skin biopsies to investigate the differences between stage IV pressure ulcers that remain stalled, stagnant, and unhealed versus those exhibiting clinical and histological signs of improvement. Our data reveal a rich diversity of proteins that are dynamically modulated, and we selectively highlight a family of calcium binding proteins (S-100 molecules) including calcyclin (S100-A6), calgranulins A (S100-A8) and B (S100-A9), and calgizzarin (S100-A11). IMS allowed us to target three discrete regions of interest: the wound bed, adjacent dermis, and hypertrophic epidermis. Plots derived using unsupervised principal component analysis of the global protein signatures within these three spatial niches indicate that these data from wound signatures have potential as a prognostic tool since they appear to delineate wounds that are favorably responding to therapeutic interventions versus those that remain stagnant or intractable in their healing status. Our discovery-based approach with IMS augments current knowledge of the molecular signatures within pressure ulcers while providing a rationale for a focused examination of the role of calcium modulators within the context of impaired wound healing.
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15 MeSH Terms
Global deletion of Ankrd1 results in a wound-healing phenotype associated with dermal fibroblast dysfunction.
Samaras SE, Almodóvar-García K, Wu N, Yu F, Davidson JM
(2015) Am J Pathol 185: 96-109
MeSH Terms: Adenoviridae, Animals, Cell Movement, Collagen, Crosses, Genetic, Exons, Female, Fibroblasts, Gels, Gene Deletion, Gene Expression Profiling, Genotype, Green Fluorescent Proteins, MAP Kinase Signaling System, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Mice, Transgenic, Muscle Proteins, Necrosis, Nuclear Proteins, Organ Size, Phenotype, Promoter Regions, Genetic, RNA, Recombination, Genetic, Repressor Proteins, Skin, Wound Healing
Show Abstract · Added January 20, 2015
The expression of ankyrin repeat domain protein 1 (Ankrd1), a transcriptional cofactor and sarcomeric component, is strongly elevated by wounding and tissue injury. We developed a conditional Ankrd1(fl/fl) mouse, performed global deletion with Sox2-cre, and assessed the role of this protein in cutaneous wound healing. Although global deletion of Ankrd1 did not affect mouse viability or development, Ankrd1(-/-) mice had at least two significant wound-healing phenotypes: extensive necrosis of ischemic skin flaps, which was reversed by adenoviral expression of ANKRD1, and delayed excisional wound closure, which was characterized by decreased contraction and reduced granulation tissue thickness. Skin fibroblasts isolated from Ankrd1(-/-) mice did not spread or migrate on collagen- or fibronectin-coated surfaces as efficiently as fibroblasts isolated from Ankrd1(fl/fl) mice. More important, Ankrd1(-/-) fibroblasts failed to contract three-dimensional floating collagen gels. Reconstitution of ANKRD1 by adenoviral infection stimulated both collagen gel contraction and actin fiber organization. These in vitro data were consistent with in vivo wound closure studies, and suggest that ANKRD1 is important for the proper interaction of fibroblasts with a compliant collagenous matrix both in vitro and in vivo.
Copyright © 2015 American Society for Investigative Pathology. Published by Elsevier Inc. All rights reserved.
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30 MeSH Terms
Examination of the Early Diagnostic Applicability of Active Dynamic Thermography for Burn Wound Depth Assessment and Concept Analysis.
Prindeze NJ, Fathi P, Mino MJ, Mauskar NA, Travis TE, Paul DW, Moffatt LT, Shupp JW
(2015) J Burn Care Res 36: 626-35
MeSH Terms: Animals, Biopsy, Needle, Burns, Disease Models, Animal, Early Diagnosis, Immunohistochemistry, Injury Severity Score, Laser-Doppler Flowmetry, Male, Random Allocation, Sensitivity and Specificity, Skin, Swine, Thermography, Wound Healing
Show Abstract · Added March 20, 2018
Despite advances in perfusion imaging, burn wound imaging technology continues to lag behind that of other fields. Quantification of blood flow is able to predict time for healing, but clear assessment of burn depth is still questionable. Active dynamic thermography (ADT) is a noncontact imaging modality capable of distinguishing tissue of different thermal conductivities. Utilizing the abnormal heat transfer properties of the burn zones, we examined whether ADT was useful in the determination of burn depth in a model of early burn wound evaluation. Duroc pigs (castrated male; n = 3) were anesthetized, and two burns were created with an aluminum billet at 3 and 12 seconds. These contact times resulted in superficial partial and deep partial thickness burn wounds, respectively. ADT and laser Doppler imaging (LDI) imaging were performed every 30 minutes postburn for a total of five imaging sessions ending 150 minutes postburn. For ADT, imaging excitation was performed for 42-120 seconds with dual quartz-infrared lamps, and subsequent infrared image capture was performed for 300 seconds. MATLAB-assisted image analysis was performed to determine burn zone region of interest thermal relaxation and characteristic patterns. LDI was performed with a moorLDI system, and biopsies were captured for histology following the 150-minute imaging session. Both ADT and LDI imaging modalities are able to detect different physical properties at 30, 60, 90 120, and 150 minutes postburn with statistical significance (P < 0.05). Resultant ADT cooling curves characterize greater differences with greater stimulation and a potentially more identifiable differential cooling characteristic. Histological analysis confirmed burn depth. This preliminary work confirms that ADT can measure burn depth and is deserving of further research either as a stand-alone imaging technology or in combination with a device to assess perfusion.
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15 MeSH Terms
Biodegradable lysine-derived polyurethane scaffolds promote healing in a porcine full-thickness excisional wound model.
Adolph EJ, Pollins AC, Cardwell NL, Davidson JM, Guelcher SA, Nanney LB
(2014) J Biomater Sci Polym Ed 25: 1973-85
MeSH Terms: Animals, Apoptosis, Biocompatible Materials, Cell Proliferation, Humans, Lysine, Macrophages, Mechanical Phenomena, Neovascularization, Physiologic, Polyurethanes, Skin, Structure-Activity Relationship, Swine, Tissue Scaffolds, Wound Healing
Show Abstract · Added February 23, 2016
Lysine-derived polyurethane scaffolds (LTI-PUR) support cutaneous wound healing in loose-skinned small animal models. Due to the physiological and anatomical similarities of human and pig skin, we investigated the capacity of LTI-PUR scaffolds to support wound healing in a porcine excisional wound model. Modifications to scaffold design included the addition of carboxymethylcellulose (CMC) as a porogen to increase interconnectivity and an additional plasma treatment (Plasma) to decrease surface hydrophobicity. All LTI-PUR scaffold and formulations supported cellular infiltration and were biodegradable. At 15 days, CMC and plasma scaffolds simulated increased macrophages more so than LTI PUR or no treatment. This response was consistent with macrophage-mediated oxidative degradation of the lysine component of the scaffolds. Cell proliferation was similar in control and scaffold-treated wounds at 8 and 15 days. Neither apoptosis nor blood vessel area density showed significant differences in the presence of any of the scaffold variations compared with untreated wounds, providing further evidence that these synthetic biomaterials had no adverse effects on those pivotal wound healing processes. During the critical phase of granulation tissue formation in full thickness porcine excisional wounds, LTI-PUR scaffolds supported tissue infiltration, while undergoing biodegradation. Modifications to scaffold fabrication modify the reparative process. This study emphasizes the biocompatibility and favorable cellular responses of PUR scaffolding formulations in a clinically relevant animal model.
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15 MeSH Terms
α2β1 Integrin.
Madamanchi A, Santoro SA, Zutter MM
(2014) Adv Exp Med Biol 819: 41-60
MeSH Terms: Animals, Hemostasis, Humans, Immunity, Innate, Integrin alpha2beta1, Neovascularization, Physiologic, Protein Structure, Tertiary, Signal Transduction, Wound Healing
Show Abstract · Added February 12, 2015
The α2β1 integrin, also known as VLA-2, GPIa-IIa, CD49b, was first identified as an extracellular matrix receptor for collagens and/or laminins [55, 56]. It is now recognized that the α2β1 integrin serves as a receptor for many matrix and nonmatrix molecules [35, 79, 128]. Extensive analyses have clearly elucidated the α2 I domain structural motifs required for ligand binding, and also defined distinct conformations that lead to inactive, partially active or highly active ligand binding [3, 37, 66, 123, 136, 137, 140]. The mechanisms by which the α2β1 integrin plays a critical role in platelet function and homeostasis have been carefully defined via in vitro and in vivo experiments [76, 104, 117, 125]. Genetic and epidemiologic studies have confirmed human physiology and disease states mediated by this receptor in immunity, cancer, and development [6, 20, 21, 32, 43, 90]. The role of the α2β1 integrin in these multiple complex biologic processes will be discussed in the chapter.
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9 MeSH Terms
The ophthalmology microscalpel versus standard scalpels and wound healing in a rat model.
Pearce EC, Hall JE, Boyd KL, Rousseau B, Ries WR
(2014) Otolaryngol Head Neck Surg 151: 424-30
MeSH Terms: Animals, Dermatologic Surgical Procedures, Disease Models, Animal, Electrosurgery, Equipment Safety, Immunohistochemistry, Ophthalmology, Random Allocation, Rats, Rats, Sprague-Dawley, Surgical Instruments, Tensile Strength, Wound Healing, Wounds and Injuries
Show Abstract · Added February 12, 2015
OBJECTIVE - We tested the hypothesis that the ophthalmology microscalpel, compared to standard incisional instruments, causes less trauma during incisions resulting in decreased inflammation and greater tensile strength of wounds.
STUDY DESIGN - Prospective animal study.
SETTING - Animal laboratory.
SUBJECTS AND METHODS - Thirty-four Sprague-Dawley rats received dorsum skin incisions with the microscalpel, electrosurgical device, 11 blade scalpel, and 15 blade scalpel. Wounds were harvested at 1 week, 2 weeks, 3 weeks, and 6 weeks, then analyzed histologically in a blinded manner for inflammation markers and tested for tensile strength.
RESULTS - The microscalpel wounds had significantly higher tensile strength compared to the 15 blade (P = .045) and electrocautery device (P = .000) but equivocal strength to the 11 blade (P = .457). The electrocautery wounds were weaker than all 3 steel blades. No significant difference was found between the microscalpel, 11 blade, and 15 blade incisions for the 5 markers of inflammation. Electrocautery wounds had significantly worse inflammatory scores, specifically, higher angiogenesis and larger wound gap compared to the microscalpel (P = .004, P = .002), 11 blade (P = .007, P = .023), and 15 blade (P = .010, P = .003), respectively.
CONCLUSION - Microscalpel incisions result in less inflammation and increased tensile strength compared with electrocautery and higher tensile strength compared to the 15 blade in the rat model. Inflammation scores were equivocal between the microscalpel, 11 blade, and 15 blade. Our findings support the use of the microscalpel blade for facial plastic and reconstructive procedures. Prospective, randomized human studies are warranted.
© American Academy of Otolaryngology—Head and Neck Surgery Foundation 2014.
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14 MeSH Terms