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Proline Precursors and Collagen Synthesis: Biochemical Challenges of Nutrient Supplementation and Wound Healing.
Albaugh VL, Mukherjee K, Barbul A
(2017) J Nutr 147: 2011-2017
MeSH Terms: Amino Acids, Animals, Collagen, Dietary Supplements, Disease Models, Animal, Humans, Proline, Wound Healing
Show Abstract · Added January 4, 2019
Wound healing is a complex process marked by highly coordinated immune fluxes into an area of tissue injury; these are required for re-establishment of normal tissue integrity. Along with this cascade of cellular players, wound healing also requires coordinated flux through a number of biochemical pathways, leading to synthesis of collagen and recycling or removal of damaged tissues. The availability of nutrients, especially amino acids, is critical for wound healing, and enteral supplementation has been intensely studied as a potential mechanism to augment wound healing-either by increasing tensile strength, decreasing healing time, or both. From a practical standpoint, although enteral nutrient supplementation may seem like a reasonable strategy to augment healing, a number of biochemical and physiologic barriers exist that limit this strategy. In this critical review, the physiology of enteral amino acid metabolism and supplementation and challenges therein are discussed in the context of splanchnic physiology and biochemistry. Additionally, a review of studies examining various methods of amino acid supplementation and the associated effects on wound outcomes are discussed.
© 2017 American Society for Nutrition.
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Multiple Mechanisms Drive Calcium Signal Dynamics around Laser-Induced Epithelial Wounds.
Shannon EK, Stevens A, Edrington W, Zhao Y, Jayasinghe AK, Page-McCaw A, Hutson MS
(2017) Biophys J 113: 1623-1635
MeSH Terms: Animals, Animals, Genetically Modified, Calcium, Calcium Signaling, Cell Membrane, Cytosol, Drosophila, Epithelial Cells, Lasers, Microscopy, Confocal, Voltage-Sensitive Dye Imaging, Wings, Animal, Wound Healing
Show Abstract · Added March 20, 2018
Epithelial wound healing is an evolutionarily conserved process that requires coordination across a field of cells. Studies in many organisms have shown that cytosolic calcium levels rise within a field of cells around the wound and spread to neighboring cells, within seconds of wounding. Although calcium is a known potent second messenger and master regulator of wound-healing programs, it is unknown what initiates the rise of cytosolic calcium across the wound field. Here we use laser ablation, a commonly used technique for the precision removal of cells or subcellular components, as a tool to investigate mechanisms of calcium entry upon wounding. Despite its precise ablation capabilities, we find that this technique damages cells outside the primary wound via a laser-induced cavitation bubble, which forms and collapses within microseconds of ablation. This cavitation bubble damages the plasma membranes of cells it contacts, tens of microns away from the wound, allowing direct calcium entry from extracellular fluid into damaged cells. Approximately 45 s after this rapid influx of calcium, we observe a second influx of calcium that spreads to neighboring cells beyond the footprint of cavitation. The occurrence of this second, delayed calcium expansion event is predicted by wound size, indicating that a separate mechanism of calcium entry exists, corresponding to cell loss at the primary wound. Our research demonstrates that the damage profile of laser ablation is more similar to a crush injury than the precision removal of individual cells. The generation of membrane microtears upon ablation is consistent with studies in the field of optoporation, which investigate ablation-induced cellular permeability. We conclude that multiple types of damage, including microtears and cell loss, result in multiple mechanisms of calcium influx around epithelial wounds.
Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.
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13 MeSH Terms
Lrig1+ gastric isthmal progenitor cells restore normal gastric lineage cells during damage recovery in adult mouse stomach.
Choi E, Lantz TL, Vlacich G, Keeley TM, Samuelson LC, Coffey RJ, Goldenring JR, Powell AE
(2018) Gut 67: 1595-1605
MeSH Terms: Animals, Biomarkers, Cell Lineage, Disease Models, Animal, Gastric Mucosa, Membrane Glycoproteins, Mice, Mice, Knockout, Nerve Tissue Proteins, Predictive Value of Tests, Sensitivity and Specificity, Stem Cells, Stomach Ulcer, Wound Healing
Show Abstract · Added September 27, 2017
OBJECTIVE - Lrig1 is a marker of proliferative and quiescent stem cells in the skin and intestine. We examined whether Lrig1-expressing cells are long-lived gastric progenitors in gastric glands in the mouse stomach. We also investigated how the Lrig1-expressing progenitor cells contribute to the regeneration of normal gastric mucosa by lineage commitment to parietal cells after acute gastric injury in mice.
DESIGN - We performed lineage labelling using (Lrig1/YFP) or (Lrig1/LacZ) mice to examine whether the Lrig1-YFP-marked cells are gastric progenitor cells. We studied whether Lrig1-YFP-marked cells give rise to normal gastric lineage cells in damaged mucosa using Lrig1/YFP mice after treatment with DMP-777 to induce acute injury. We also studied Lrig1- (Lrig1 knockout) mice to examine whether the Lrig1 protein is required for regeneration of gastric corpus mucosa after acute injury.
RESULTS - Lrig1-YFP-marked cells give rise to gastric lineage epithelial cells both in the gastric corpus and antrum, in contrast to published results that Lgr5 only marks progenitor cells within the gastric antrum. Lrig1-YFP-marked cells contribute to replacement of damaged gastric oxyntic glands during the recovery phase after acute oxyntic atrophy in the gastric corpus. Lrig1 null mice recovered normally from acute gastric mucosal injury indicating that Lrig1 protein is not required for lineage differentiation. Lrig1+ isthmal progenitor cells did not contribute to transdifferentiating chief cell lineages after acute oxyntic atrophy.
CONCLUSIONS - Lrig1 marks gastric corpus epithelial progenitor cells capable of repopulating the damaged oxyntic mucosa by differentiating into normal gastric lineage cells in mouse stomach.
© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.
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14 MeSH Terms
Porcine Ischemic Wound-Healing Model for Preclinical Testing of Degradable Biomaterials.
Patil P, Martin JR, Sarett SM, Pollins AC, Cardwell NL, Davidson JM, Guelcher SA, Nanney LB, Duvall CL
(2017) Tissue Eng Part C Methods 23: 754-762
MeSH Terms: Animals, Biocompatible Materials, Blood Vessels, Disease Models, Animal, Ischemia, Macrophages, Materials Testing, Skin, Surgical Flaps, Sus scrofa, Tissue Scaffolds, Wound Healing
Show Abstract · Added March 14, 2018
Impaired wound healing that mimics chronic human skin pathologies is difficult to achieve in current animal models, hindering testing and development of new therapeutic biomaterials that promote wound healing. In this article, we describe a refinement and simplification of the porcine ischemic wound model that increases the size and number of experimental sites per animal. By comparing three flap geometries, we adopted a superior configuration (15 × 10 cm) that enabled testing of twenty 1 cm wounds in each animal: 8 total ischemic wounds within 4 bipedicle flaps and 12 nonischemic wounds. The ischemic wounds exhibited impaired skin perfusion for ∼1 week. To demonstrate the utility of the model for comparative testing of tissue regenerative biomaterials, we evaluated the healing process in wounds implanted with highly porous poly (thioketal) urethane (PTK-UR) scaffolds that were fabricated through reaction of reactive oxygen species (ROS)-cleavable PTK macrodiols with isocyanates. PTK-lysine triisocyanate (LTI) scaffolds degraded significantly in vitro under both oxidative and hydrolytic conditions whereas PTK-hexamethylene diisocyanate trimer (HDIt) scaffolds were resistant to hydrolytic breakdown and degraded exclusively through an ROS-dependent mechanism. Upon placement into porcine wounds, both types of PTK-UR materials fostered new tissue ingrowth over 10 days in both ischemic and nonischemic tissue. However, wound perfusion, tissue infiltration and the abundance of pro-regenerative, M2-polarized macrophages were markedly lower in ischemic wounds independent of scaffold type. The PTK-LTI implants significantly improved tissue infiltration and perfusion compared with analogous PTK-HDIt scaffolds in ischemic wounds. Both LTI and HDIt-based PTK-UR implants enhanced M2 macrophage activity, and these cells were selectively localized at the scaffold/tissue interface. In sum, this modified porcine wound-healing model decreased animal usage, simplified procedures, and permitted a more robust evaluation of tissue engineering materials in preclinical wound healing research. Deployment of the model for a relevant biomaterial comparison yielded results that support the use of the PTK-LTI over the PTK-HDIt scaffold formulation for future advanced therapeutic studies.
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12 MeSH Terms
Local Delivery of PHD2 siRNA from ROS-Degradable Scaffolds to Promote Diabetic Wound Healing.
Martin JR, Nelson CE, Gupta MK, Yu F, Sarett SM, Hocking KM, Pollins AC, Nanney LB, Davidson JM, Guelcher SA, Duvall CL
(2016) Adv Healthc Mater 5: 2751-2757
MeSH Terms: Animals, Cell Proliferation, Diabetes Mellitus, Male, Neovascularization, Physiologic, Procollagen-Proline Dioxygenase, RNA, Small Interfering, Rats, Rats, Sprague-Dawley, Reactive Oxygen Species, Tissue Engineering, Tissue Scaffolds, Wound Healing
Show Abstract · Added March 14, 2018
Small interfering RNA (siRNA) delivered from reactive oxygen species-degradable tissue engineering scaffolds promotes diabetic wound healing in rats. Porous poly(thioketal-urethane) scaffolds implanted in diabetic wounds locally deliver siRNA that inhibits the expression of prolyl hydroxylase domain protein 2, thereby increasing the expression of progrowth genes and increasing vasculature, proliferating cells, and tissue development in diabetic wounds.
© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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13 MeSH Terms
Clinical investigation of biofilm in non-healing wounds by high resolution microscopy techniques.
Hurlow J, Blanz E, Gaddy JA
(2016) J Wound Care 25 Suppl 9: S11-22
MeSH Terms: Adolescent, Anti-Infective Agents, Local, Bandages, Biofilms, Debridement, Humans, Microscopy, Wound Healing, Wound Infection
Show Abstract · Added April 26, 2017
OBJECTIVE - The aim of this study was to analyse wound biofilm from a clinical perspective. Research has shown that biofilm is the preferred microbial phenotype in health and disease and is present in a majority of chronic wounds. Biofilm has been linked to chronic wound inflammation, impairment in granulation tissue and epithelial migration, yet there lacks the ability to confirm the clinical presence of biofilm. This study links the clinical setting with microscopic laboratory confirmation of the presence of biofilm in carefully selected wound debridement samples.
METHOD - Human wound debridement samples were collected from adult patients with chronic non-healing wounds who presented at the wound care centre. Sample choice was guided by an algorithm that was developed based on what is known about the characteristics of wound biofilm. The samples were then evaluated by light microscopy and scanning electron microscopy for the presence of biofilm. Details about subject history and treatment were recorded. Adherence to biofilm-based wound care (BBWC) strategies was inconsistent. Other standard antimicrobial dressings were used and no modern antiseptic wound dressings with the addition of proven antibiofilm agents were available for use.
RESULTS - Of the patients recruited, 75% of the macroscopic samples contained biofilm despite the prior use of modern antiseptic wound dressings and in some cases, systemic antibiotics. Wounds found to contain biofilm were not all acutely infected but biofilm was present when infection was noted. The clinical histories associated with positive samples were consistent with ideas presented in the algorithm used to guide sample selection.
CONCLUSION - Visual cues can be used by the clinician to guide suspicion of the presence of wound biofilm. This suspicion can be further enhanced with the use of a clinical algorithm. Standard antiseptic wound dressings used in this study demonstrated limited antibiofilm efficacy. This study also highlighted a need for the clinical team to focus on expiration of dressing action and consistent practice of BBWC strategies which includes the use of proven antibiofilm agents.
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TLR3 Agonist Poly-IC Induces IL-33 and Promotes Myelin Repair.
Natarajan C, Yao SY, Sriram S
(2016) PLoS One 11: e0152163
MeSH Terms: Animals, Cell Differentiation, Cell Nucleus, Cell Polarity, Cells, Cultured, Corpus Callosum, Enzyme Activation, Intercellular Signaling Peptides and Proteins, Interleukin-33, Lysophosphatidylcholines, Macrophages, Models, Biological, Myelin Basic Protein, Myelin Sheath, Neuroglia, Oligodendroglia, Phenotype, Phosphorylation, Poly I-C, Rats, Sprague-Dawley, Real-Time Polymerase Chain Reaction, Recombinant Proteins, Stem Cells, Toll-Like Receptor 3, Transcription, Genetic, Up-Regulation, Wound Healing, p38 Mitogen-Activated Protein Kinases
Show Abstract · Added April 18, 2017
BACKGROUND - Impaired remyelination of demyelinated axons is a major cause of neurological disability. In inflammatory demyelinating disease of the central nervous system (CNS), although remyelination does happen, it is often incomplete, resulting in poor clinical recovery. Poly-IC a known TLR3 agonist and IL-33, a cytokine which is induced by poly-IC are known to influence recovery and promote repair in experimental models of CNS demyelination.
METHODOLOGY AND PRINCIPAL FINDINGS - We examined the effect of addition of poly-IC and IL-33 on the differentiation and maturation of oligodendrocyte precursor cells (OPC) cultured in vitro. Both Poly-IC and IL-33 induced transcription of myelin genes and the differentiation of OPC to mature myelin forming cells. Poly-IC induced IL-33 in OPC and addition of IL-33 to in vitro cultures, amplified further, IL-33 expression suggesting an autocrine regulation of IL-33. Poly-IC and IL-33 also induced phosphorylation of p38MAPK, a signaling molecule involved in myelination. Following the induction of gliotoxic injury with lysolecithin to the corpus callosum (CC), treatment of animals with poly-IC resulted in greater recruitment of OPC and increased staining for myelin in areas of demyelination. Also, poly-IC treated animals showed greater expression of IL-33 and higher expression of M2 phenotype macrophages in the CC.
CONCLUSION/SIGNIFICANCE - Our studies suggest that poly-IC and IL-33 play a role in myelin repair by enhancing expression of myelin genes and are therefore attractive therapeutic agents for use as remyelinating agents in human demyelinating disease.
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28 MeSH Terms
Augmenting Surgery via Multi-scale Modeling and Translational Systems Biology in the Era of Precision Medicine: A Multidisciplinary Perspective.
Kassab GS, An G, Sander EA, Miga MI, Guccione JM, Ji S, Vodovotz Y
(2016) Ann Biomed Eng 44: 2611-25
MeSH Terms: Cardiac Surgical Procedures, Computer Simulation, Humans, Patient-Specific Modeling, Precision Medicine, Translational Medical Research, Wound Healing, Wounds and Injuries
Show Abstract · Added July 23, 2018
In this era of tremendous technological capabilities and increased focus on improving clinical outcomes, decreasing costs, and increasing precision, there is a need for a more quantitative approach to the field of surgery. Multiscale computational modeling has the potential to bridge the gap to the emerging paradigms of Precision Medicine and Translational Systems Biology, in which quantitative metrics and data guide patient care through improved stratification, diagnosis, and therapy. Achievements by multiple groups have demonstrated the potential for (1) multiscale computational modeling, at a biological level, of diseases treated with surgery and the surgical procedure process at the level of the individual and the population; along with (2) patient-specific, computationally-enabled surgical planning, delivery, and guidance and robotically-augmented manipulation. In this perspective article, we discuss these concepts, and cite emerging examples from the fields of trauma, wound healing, and cardiac surgery.
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Substrate modulus of 3D-printed scaffolds regulates the regenerative response in subcutaneous implants through the macrophage phenotype and Wnt signaling.
Guo R, Merkel AR, Sterling JA, Davidson JM, Guelcher SA
(2015) Biomaterials 73: 85-95
MeSH Terms: Animals, Cells, Cultured, Collagen, Down-Regulation, Fibroblasts, Humans, Intercellular Signaling Peptides and Proteins, Kinetics, Macrophages, Male, Neovascularization, Pathologic, Phenotype, Porosity, Pressure, Printing, Three-Dimensional, Rats, Rats, Sprague-Dawley, Regeneration, Tissue Engineering, Tissue Scaffolds, Wnt Proteins, Wnt Signaling Pathway, Wound Healing, beta Catenin
Show Abstract · Added February 23, 2016
The growing need for therapies to treat large cutaneous defects has driven recent interest in the design of scaffolds that stimulate regenerative wound healing. While many studies have investigated local delivery of biologics as a restorative approach, an increasing body of evidence highlights the contribution of the mechanical properties of implanted scaffolds to wound healing. In the present study, we designed poly(ester urethane) scaffolds using a templated-Fused Deposition Modeling (t-FDM) process to test the hypothesis that scaffolds with substrate modulus comparable to that of collagen fibers enhance a regenerative versus a fibrotic response. We fabricated t-FDM scaffolds with substrate moduli varying from 5 to 266 MPa to investigate the effects of substrate modulus on healing in a rat subcutaneous implant model. Angiogenesis, cellular infiltration, collagen deposition, and directional variance of collagen fibers were maximized for wounds treated with scaffolds having a substrate modulus (Ks = 24 MPa) comparable to that of collagen fibers. The enhanced regenerative response in these scaffolds was correlated with down-regulation of Wnt/β-catenin signaling in fibroblasts, as well as increased polarization of macrophages toward the restorative M2 phenotype. These observations highlight the substrate modulus of the scaffold as a key parameter regulating the regenerative versus scarring phenotype in wound healing. Our findings further point to the potential use of scaffolds with substrate moduli tuned to that of the native matrix as a therapeutic approach to improve cutaneous healing.
Copyright © 2015 Elsevier Ltd. All rights reserved.
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24 MeSH Terms
Injected biodegradable polyurethane scaffolds support tissue infiltration and delay wound contraction in a porcine excisional model.
Adolph EJ, Guo R, Pollins AC, Zienkiewicz K, Cardwell N, Davidson JM, Guelcher SA, Nanney LB
(2016) J Biomed Mater Res B Appl Biomater 104: 1679-1690
MeSH Terms: Absorbable Implants, Animals, Biodegradable Plastics, Disease Models, Animal, Polyurethanes, Swine, Tissue Scaffolds, Wound Healing, Wounds and Injuries
Show Abstract · Added February 23, 2016
The filling of wound cavities with new tissue is a challenge. We previously reported on the physical properties and wound healing kinetics of prefabricated, gas-blown polyurethane (PUR) scaffolds in rat and porcine excisional wounds. To address the capability of this material to fill complex wound cavities, this study examined the in vitro and in vivo reparative characteristics of injected PUR scaffolds employing a sucrose porogen. Using the porcine excisional wound model, we compared reparative outcomes to both preformed and injected scaffolds as well as untreated wounds at 9, 13, and 30 days after scaffold placement. Both injected and preformed scaffolds delayed wound contraction by 19% at 9 days and 12% at 13 days compared to nontreated wounds. This stenting effect proved transient since both formulations degraded by day 30. Both types of scaffolds significantly inhibited the undesirable alignment of collagen and fibroblasts through day 13. Injected scaffolds were highly compatible with sentinel cellular events of normal wound repair cell proliferation, apoptosis, and blood vessel density. The present study provides further evidence that either injected or preformed PUR scaffolds facilitate wound healing, support tissue infiltration and matrix production, delay wound contraction, and reduce scarring in a clinically relevant animal model, which underscores their potential utility as a void-filling platform for large cutaneous defects. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 104B: 1679-1690, 2016.
© 2015 Wiley Periodicals, Inc.
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