Stage Of Cutaneous Wound Healing Research Articles

The Role of TNF-Alpha in the Wound Healing Process: Molecular and Clinical Perspectives - A Systematic Literature Review

Wounds are conditions of damage and partial loss of body tissue caused by physical trauma, sharp objects, blunt objects, temperature fluctuations, chemical exposure. Wound healing is a complex interaction between cells, cytokines, mediators, and blood vessels as a natural physiological response to tissue damage. TNF-α has a crucial role in the wound healing process, facilitating body immunity, immune cell mobilization, fibroblast cell proliferation, keratinocytes, and growth factor expression. Although TNF-alpha has benefits in the early stages of wound healing, excessively high levels or a sustained inflammatory response can lead to problems, such as excessive scar tissue formation or blockages, which can impede the wound healing process. This study utilized the PRISMA-ScR (Scoping Review) protocol. A review was conducted of articles discussing the Role of TNF-Alpha in the Wound Healing Process: Molecular and Clinical Perspectives within the last 10 years. Literature search in this study using PubMed, ScienceDirect, Elsivier and Google Scolar databases Search using English, abstract and full text. After searching with the keywords: The Role of TNF-Alpha, 19,022 articles were found. The search continued using the keyword The Role of TNF-Alpha in the Wound Healing found 346 articles. The last search using the keyword The Role of TNF-Alpha in the Wound Healing: Molecular and Clinical found 26 articles. TNF-α plays an important role in the early stages of cutaneous wound healing, significantly delayed after day 3, but not by day 7. TNF-α has a crucial role in the early stages of skin wound healing, and its administration is recognized to have a positive impact on the healing response, which may pave the way for innovative strategies to address chronic skin wound healing problems.

Enhancing cutaneous wound healing: A study on the beneficial effects of nano-gelatin scaffold in rat models.

The challenges in achieving optimal outcomes for wound healing have persisted for decades, prompting ongoing exploration of interventions and management strategies. This study focuses on assessing the potential benefits of implementing a nano-gelatin scaffold for wound healing. Using a rat skin defect model, full-thickness incisional wounds were created on each side of the thoracic-lumbar regions after anesthesia. The wounds were left un-sutured, with one side covered by a gelatin nano-fibrous membrane and the other left uncovered. Wound size changes were measured on days 1, 4, 7, and 14, and on day 14, rats were sacrificed for tissue sample excision, examined with hematoxylin and eosin, and Masson's trichrome stain. Statistical comparisons were performed. The gelatin nanofibers exhibited a smooth surface with a fiber diameter of 260 ± 40 nm and porous structures with proper interconnectivity. Throughout the 14-day experimental period, significant differences in the percentage of wound closure were observed between the groups. Histological scores were higher in the experiment group, indicating less inflammation but dense and well-aligned collagen fiber formation. A preliminary clinical trial on diabetic ulcers also demonstrated promising results. This study highlights the potential of the nano-collagen fibrous membrane to reduce inflammatory infiltration and enhance fibroblast differentiation into myofibroblasts during the early stages of cutaneous wound healing. The nano-fibrous collagen membrane emerges as a promising candidate for promoting wound healing, with considerable potential for future therapeutic applications.

KY19382 Accelerates Cutaneous Wound Healing via Activation of the Wnt/β-Catenin Signaling Pathway.

The Wnt/β-catenin signaling pathway plays important roles in the multi-phases of wound healing: homeostasis, inflammation, proliferative, and remodeling phases. However, there are no clinically available therapeutic agents targeting the Wnt/β-catenin pathway. In this study, we tested the effect of 5, 6-dichloroindirubin-3'-methoxime (KY19382), a small molecule that activates the Wnt/β-catenin pathway via interference with the function of the negative feedback regulator CXXC5, on cutaneous wound healing. KY19382 significantly enhanced cell migration of human keratinocytes and dermal fibroblasts with increased levels of β-catenin, phalloidin, Keratin 14, proliferating cell nuclear antigen (PCNA), Collagen I, and alpha-smooth muscle actin (α-SMA) by activating the Wnt/β-catenin signaling pathway without causing significant cytotoxicity. In addition, levels of Collagen I, Keratin 14, PCNA, and stem cell markers were significantly increased by KY19382 in a cutaneous murine wound healing model. Moreover, KY19382 treatment accelerated re-epithelialization and neo-epidermis formation with collagen deposition and stem cell activation at an early stage of cutaneous wound healing. Overall, KY19382 accelerates wound healing via activating the Wnt/β-catenin pathway, and may have the potential to be used for the development of a new wound healing agent.

Fibrillar biopolymer-based scaffolds to study macrophage-fibroblast crosstalk in wound repair.

Controlled wound healing requires a temporal and spatial coordination of cellular activities within the surrounding extracellular matrix (ECM). Disruption of cell-cell and cell-matrix communication results in defective repair, like chronic or fibrotic wounds. Activities of macrophages and fibroblasts crucially contribute to the fate of closing wounds. To investigate the influence of the ECM as an active part controlling cellular behavior, coculture models based on fibrillar 3D biopolymers such as collagen have already been successfully used. With well-defined biochemical and biophysical properties such 3D scaffolds enable in vitro studies on cellular processes including infiltration and differentiation in an in vivo like microenvironment. Further, paracrine and autocrine signaling as well as modulation of soluble mediator transport inside the ECM can be modeled using fibrillar 3D scaffolds. Herein, we review the usage of these scaffolds in in vitro coculture models allowing in-depth studies on the crosstalk between macrophages and fibroblasts during different stages of cutaneous wound healing. A more accurate mimicry of the various processes of cellular crosstalk at the different stages of wound healing will contribute to a better understanding of the impact of biochemical and biophysical environmental parameters and help to develop further strategies against diseases such as fibrosis.

Topical formulations containing Copaifera duckei Dwyer oleoresin improve cutaneous wound healing.

Evaluation of the healing and toxicological effects of Copaifera duckei Dwyer oleoresin (CDO). Rodents with skin lesions were divided into nine groups, including daily treatments with 1, 3 and 10% CDO, collagenase, antibiotic ointment and control groups, for 14 days. Treatment with 10% CDO reduced skin edema and hyperplasia, demonstrating anti-inflammatory effect of the oil. Reduction in the wound area was observed, indicating the healing effect of CDO. Histopathological analysis showed increases in angiogenesis and re-epithelialization in animals treated with the highest concentration. On the other hand, no alterations in ulcerations, inflammatory infiltrate, hemorrhage, congestion, degeneration, percentage of collagen fibers, number of cells stained with anti-macrophage migration inhibitory factor, or density of area stained with anti-collagen I and III were found. Toxicogenetic analysis revealed no differences in micronucleus frequencies or in the ratio of polychromatic erythrocytes to total erythrocytes between treated and negative control, demonstrating the absence of genotoxicity and cytotoxicity, respectively. There was no difference in levels of liver enzymes among groups, indicating the absence of hepatotoxicity. Formulations of CDO exerted beneficial effects on the stages of cutaneous wound healing and are promising options for the treatment of wounds.

Research Advances in the Application of Adipose-Derived Stem Cells Derived Exosomes in Cutaneous Wound Healing.

Cutaneous wound healing has always been an intractable medical problem for both clinicians and researchers, with an urgent need for more efficacious methods to achieve optimal outcomes morphologically and functionally. Stem cells, the body's rapid response ‘road repair crew,’ being on standby to combat tissue injuries, are an essential part of regenerative medicine. Currently, the use of adipose-derived stem cells (ADSCs), a kind of mesenchymal stem cells with multipotent differentiation and self-renewal capacity, is surging in the field of cutaneous wound healing. ADSCs may exert influences either by releasing paracrine signalling factors or differentiating into mature adipose cells to provide the ‘building blocks’ for engineered tissue. As an important paracrine substance released from ADSCs, exosomes are a kind of extracellular vesicles and carrying various bioactive molecules mediating adjacent or distant intercellular communication. Previous studies have indicated that ADSCs derived exosomes (ADSCs-Exos) promoted skin wound healing by affecting all stages of wound healing, including regulating inflammatory response, promoting proliferation and migration of fibroblasts or keratinocytes, facilitating angiogenesis, and regulating remodeling of extracellular matrix, which have provided new opportunities for understanding how ADSCs-Exos mediate intercellular communication in pathological processes of the skin and therapeutic strategies for cutaneous wound repair. In this review, we focus on elucidating the role of ADSCs-Exos at various stages of cutaneous wound healing, detailing the latest developments, and presenting some challenges necessary to be addressed in this field, with the expectation of providing a new perspective on how to best utilize this powerful cell-free therapy in the future.

Phospholipase Cε deficiency delays the early stage of cutaneous wound healing and attenuates scar formation in mice

This study aimed to investigate the role of phospholipase Cε (PLCε) in the skin wound healing process. PLCε, an effect factor of Ras/Rap small G protein, plays a crucial role in skin inflammation by regulating inflammatory cytokines. Inflammatory responses are closely associated with wound healing. Full-thickness skin wounds were made in the PLCε knockout (KO) and wild-type (WT) mice, and the healing process was analyzed. The macroscopic wound closure rate declined in the PLCε KO mice on days 3, 4, and 5 after wounding, following the decreased expression of interleukin (IL)-6, chemokine (C-X-C motif) ligand (Cxcl)-1, Cxcl-2, and chemokine (C-C motif) ligand (Ccl) 20. The proliferation rate of epidermal keratinocytes was not affected by PLCε, but silencing of PLCε resulted in the delayed migration of keratinocytes. Moreover, the scars were found to be much smaller in the PLCε KO mice than in the WT mice. The mRNA expression of Ccl20, collagen (Col) 6a1, and Col17a1 decreased in the PLCε KO mice. These results were in agreement with a previous hypothesis that PLCε might delay the early stage of cutaneous wound healing by inhibiting the migration of keratinocytes, and decrease the expression of Col6a1, Col17a1, and Ccl20 by inhibiting the inflammatory response to reduce scar formation. This study shed light on a novel role of PLCε in wound healing and provided new therapeutic approaches to target PLCε for diminishing scar formation after injury.

Activities of MSCs Derived from Transgenic Mice Seeded on ADM Scaffolds in Wound Healing and Assessment by Advanced Optical Techniques

Background/Aims: Bone marrow Mesenchymal stem cells (MSCs) are promising for promoting cutaneous wound healing through reinforcing cellular processes. We evaluated the effect of GFP-tagged MSCs transplantation on skin regeneration in excisional wounds in mice. Methods: MSCs from GFP-labeled transgenic mice were co-cultured with acellular dermal matrix (ADM) scaffolds, and MSC-ADM scaffolds were transplanted into surgical skin wounds of BALB/c mice. After implantation, the survival and behavior of MSCs were examined by second harmonic generation and two-photon excitation fluorescence imaging, western blotting and DNA amplification and sequencing. Results: GFP-tagged MSCs were retained inside the regenerating skin until day 14 post-transplantation. Alpha-smooth muscle actin (α-SMA) and vimentin (VIM) were detected at 3, 5, 7, and 14 days post-transplantation by immunofluorescence double labeling. Although the GFP⁺/α-SMA⁺- and GFP⁺/VIM⁺-cell numbers decreased gradually with healing time, α-SMA⁺- and VIM⁺-cell numbers significantly increased, most of them were endogenous functional cells which were related to angiogenesis and collagen fiber structural remodeling. Conclusion: Therefore, in the initial stage of wound healing, transplanted MSCs differentiated into functional cells and played paracrine roles to recruit more endogenous cells for tissue remodeling. With the disappearance of exogenous cells, endogenous cells were responsible for the latter stage of cutaneous wound healing.

Laser phototherapy improves early stage of cutaneous wound healing of rats under hyperlipidic diet.

The aim of this study was to evaluate the influence of laser photobiomodulation in cutaneous healing of rats under a hyperlipidic diet. Forty-eight Wistar Albinus rats, weaned, received standard diet (SD) or hyperlipidic diet (HD) for 20weeks. The groups were divided into SD rats and HD rats, SD-irradiated rats (LSD), and HD-irradiated rats (LHD). Standard cutaneous wound (1cm(2)) was created on the dorsum of each rat. The irradiation started immediately after surgery and every 48h for 7 or 14days (λ660nm, 40mW, 6J/cm(2), ϕ 0,04cm(2), CW), when they were killed under deep anesthesia. The specimens were removed, routinely processed, stained with hematoxylin/eosin (H/E), and evaluated by light microscopy. Rats fed with hyperlipidic diet had greater intensity in the inflammatory process and prolonged hyperemia. At day 7, the intensity of inflammation was reduced in LSD and LHD groups when compared to their control groups, SD (p = 0.002) and HD (p = 0.02). There was an increase in fibroblast proliferation and collagen deposition, especially in the LHD group. At day 14, the HD group presented more intensive hyperemia than the SD group. It can be concluded that the hyperlipidic diet modified the inflammation pattern in wound healing and that laser light has a positive biomodulative effect on the healing process only in early stages.

The Role of Neuromediators and Innervation in Cutaneous Wound Healing.

The skin is densely innervated with an intricate network of cutaneous nerves, neuromediators and specific receptors which influence a variety of physiological and disease processes. There is emerging evidence that cutaneous innervation may play an important role in mediating wound healing. This review aims to comprehensively examine the evidence that signifies the role of innervation during the overlapping stages of cutaneous wound healing. Numerous neuropeptides that are secreted by the sensory and autonomic nerve fibres play an essential part during the distinct phases of wound healing. Delayed wound healing in diabetes and fetal cutaneous regeneration following wounding further highlights the pivotal role skin innervation and its associated neuromediators play in wound healing. Understanding the mechanisms via which cutaneous innervation modulates wound healing in both the adult and fetus will provide opportunities to develop therapeutic devices which could manipulate skin innervation to aid wound healing.

Combined Mulberry Leaf and Fruit Extract Improved Early Stage of Cutaneous Wound Healing in High-Fat Diet-Induced Obese Mice.

Obesity is a pandemic that causes many health challenges, including difficulties in achieving proper wound healing without complications. The current study investigated the role of NLRP3 inflammasome in the early stages of cutaneous wound healing and the effect of combined mulberry leaf and fruit extract (MLFE) on cutaneous NLRP inflammasome involvement in delayed wound healing mice with high-fat diet (HFD)-induced obesity. After obesity was induced by HFD for 10 weeks, the mice were supplemented with MLFE (at a dose of 500 mg/kg containing 333.3 mg/kg of mulberry leaf extract and 166.7 mg/kg of mulberry fruit extract) by gavage, 5 days/week for 12 weeks. MLFE supplementation ameliorated delayed wound closure in obese mice. While wound size was positively correlated with fasting blood glucose level during the early stage of wound healing, it was strongly correlated with body weight gain and body fat mass during the later stage of wound healing. Under obese conditions, the levels of NLRP3 inflammasome and its related markers (pro-caspase-1 and precursor/mature interleukin 1 beta) were increased at a basal level, but the NLRP3 inflammasome was suppressed during the inflammatory stage of cutaneous wound healing. However, MLFE supplementation stimulated cutaneous NLRP3 inflammasome in HFD-induced obese mice (day 3). Taken together, stimulating the NLRP3 inflammasome might be beneficial in the early inflammatory stage of cutaneous wound healing and MLFE could be a potential therapeutic intervention in delayed wound healing through activation of the NLRP3 inflammasome in obesity.

Complement deficiency promotes cutaneous wound healing in mice.

Wound healing is a complex homeostatic response to injury that engages numerous cellular activities, processes, and cell-to-cell interactions. The complement system, an intricate network of proteins with important roles in immune surveillance and homeostasis, has been implicated in many physiological processes; however, its role in wound healing remains largely unexplored. In this study, we employ a murine model of excisional cutaneous wound healing and show that C3(-/-) mice exhibit accelerated early stages of wound healing. Reconstitution of C3(-/-) mice with serum from C3(+/+) mice or purified human C3 abrogated the accelerated wound-healing phenotype. Wound histology of C3(-/-) mice revealed a reduction in inflammatory infiltrate compared with C3(+/+) mice. C3 deficiency also resulted in increased accumulation of mast cells and advanced angiogenesis. We further show that mice deficient in the downstream complement effector C5 exhibit a similar wound-healing phenotype, which is recapitulated in C5aR1(-/-) mice, but not C3aR(-/-) or C5aR2(-/-) mice. Taken together, these data suggest that C5a signaling through C5aR may in part play a pivotal role in recruitment and activation of inflammatory cells to the wound environment, which in turn could delay the early stages of cutaneous wound healing. These findings also suggest a previously underappreciated role for complement in wound healing, and may have therapeutic implications for conditions of delayed wound healing.

Dietary Supplementation of Genistein Attenuates Inflammatory Responses and Oxidative Stress during Cutaneous Wound Healing in Diabetic Mice

Delayed wound healing is one of the most serious diabetic complications. This is mainly due to increased impairment of inflammatory response and oxidative stress in diabetic tissues. In this study, we investigated whether genistein supplementation attenuated inflammatory response and oxidative stress during the early stage of cutaneous wound healing in alloxan-induced diabetic mice. Diabetes was induced by administration of alloxan monohydrate. Mice with diabetes [fasting blood glucose (FBG) levels >= 250 mg/dl] were fed AIN-93G rodent diet contained 0%, 0.025% or 0.1% genistein. Diabetic mice showed increased hepatic MDA concentration, delayed wound closure rate, and dysregulated hepatic antioxidant enzyme expression compared to nondiabetic mice. However, genistein supplementation decreased hepatic MDA levels, accelerated wound closure rate, and rectified hepatic antioxidant enzymes expression under diabetic condition without regulation of FBG levels. Moreover, diabetic mice had excessive inflammatory responses such as increases in NFkB, pIkB, TNFa, COX-2, and iNOS and dysregulation of the antioxidant defense system demonstrated by Nrf2, HO-1 and MnSOD during the inflammatory phase of cutaneous wound healing, contrary to nondiabetic mice. However, genistein supplementation accelerated wound closure rate by modulation of both inflammatory response and oxidative stress-related markers in diabetic mice. Taken together, we suggest that genistein would be a potential therapeutic in prevention and treatment of delayed wound healing in diabetic patients by regulation of inflammatory response and oxidative stress.

Interfacial growth during closure of a cutaneous wound: stress generation and wrinkle formation.

A biomechanical growth model for the proliferation stage of cutaneous wound healing is developed emphasizing the emergence of stress and wrinkled skin during the healing process. The healing is assumed to be primarily driven by growth at the wound edge (i.e. the interface between the wound and the skin) leading to incompatible growth strains. A closed form solution of the boundary value problem is obtained using a Varga hyperelastic membrane model for both the skin and the wound. The nature of the solution is explored for various parametric values of the skin tension, healing rate, edge incompatibility, wrinkled region radius, and wound stiffness. The obtained results for the stress field, wrinkling, and rate of healing are qualitatively in good agreement with the existing experimental observations.

Growth promoting substrates for human dermal fibroblasts provided by artificial extracellular matrices composed of collagen I and sulfated glycosaminoglycans

The application of native extracellular matrix (ECM) components is a promising approach for biomaterial design. Here, we investigated artificial ECM (aECM) consisting of collagen I (coll) and the glycosaminoglycans (GAGs) hyaluronan (HA) or chondroitin sulfate (CS). Additionally, GAGs were chemically modified by the introduction of sulfate groups to obtain low-sulfated and high-sulfated GAG derivatives. Sulfate groups are expected to bind and concentrate growth factors and improve their bioactivity. In this study we analyzed the effect of aECM on initial adhesion, proliferation, ECM synthesis and differentiation of human dermal fibroblasts (dFb) within 8–48 h. We show that initial adhesion and cell proliferation of dFb progressively increased in a sulfate dependent manner. In contrast, synthesis of ECM components coll and HA was decreased on high-sulfated aECM coll/HA3.0 and coll/CS3.1. Furthermore, the matrix metallo-proteinase-1 (MMP-1) was down-regulated on coll/HA3.0 and coll/CS3.1 on mRNA and protein level. The fibroblast differentiation marker α-smooth muscle actin (αSMA) is not affected by aECM on mRNA level. Artificial ECM consisting of coll and high-sulfated GAGs proves to be a suitable biomaterial for dFb adhesion and proliferation that induces a “proliferative phenotype” of dFb found in the early stages of cutaneous wound healing.

Design of Smart Nanofilms for Regenerative Medicine

Wound healing can be classified into several categories based their severity. There are many modalities to achieve wound healing but, the significant variety in wound types fosters a need to develop more advanced modalities that accelerate the wound healing process. This project developed a smart nanofilm using biocompatible polyelectrolyte nanofilm multilayers (PNMs) with embedded growth factors. PNMs were achieved using nebulization, an expedient tool for nanoassembly. Nebulization significantly reduces deposition time and does note compromise nanofilm characteristics. Oppositely charged PNMs, Chondroitin Sulfate (CS), Protamine Sulfate (PS), Poly‐L‐lysine (PLL) and Poly‐Glutamic acid (PGA), served as effective nanoarchitectures. PNMs (with/without platelet derived growth factor (PDGF) were formed on organtypic keratinocyte cultures (with/without wound creation). PNMs were assembled with PDGF as an active component in deep, double layered, or superficial ten‐layer architectures. AFM analysis demonstrated that heightened surface roughness and thickness is achieved with the nebulization technique when compared to the nanoassembly. Nanofilms further enhanced cell adhesion, proliferation and protein synthesis. The study suggests that bioactive, PNMs, acting as a local drug delivery system and dressing can enhance the initial stages of cutaneous wound healing.Grant Funding Source: National Science Foundation

Effect of Dietary Conjugated Linoleic Acid Supplementation on Early Inflammatory Responses during Cutaneous Wound Healing

Inflammatory response is considered the most important period that regulates the entire healing process. Conjugated linoleic acid (CLA), a class of linoleic acid positional and geometric isomers, is well known for its antioxidant and anti-inflammatory properties. We hypothesized that dietary CLA supplementation accelerates cutaneous wound healing by regulating antioxidant and anti-inflammatory functions. To investigate wound closure rates and inflammatory responses, we used a full-thickness excisional wound model after 2-week treatments with control, 0.5%, or 1% CLA-supplemented diet. Mice fed dietary CLA supplementation had reduced levels of oxidative stress and inflammatory markers. Moreover, the wound closure rate was improved significantly in mice fed a 1% CLA-supplemented diet during early stage of wound healing (inflammatory stage). We conclude that dietary CLA supplementation enhances the early stage of cutaneous wound healing as a result of modulating oxidative stress and inflammatory responses.

Lipid Mediators in Skin Inflammation: Updates and Current Views

Lipid Mediators in Skin Inflammation: Updates and Current Views

Dietary Zinc Alters Early Inflammatory Responses during Cutaneous Wound Healing in Weanling CD-1 Mice

Zinc deficiency is a well-known health problem associated with delayed wound healing, yet the precise mechanisms that underlie the delay remain unknown. We hypothesized that zinc deficiency delays wound healing as a result of decreased nuclear factor (NF)kappaB activation, reduced expression of proinflammatory cytokines [interleukin (IL)-1beta and tumor necrosis factor (TNF)-alpha], and a decrease in neutrophil infiltration during the early stage of cutaneous wound healing. We used a cutaneous, full-thickness excisional wound model in CD-1 mice to examine the rate of wound closure as well as mRNA levels of inhibitory (I)kappaBalpha, IL-1beta, and TNF-alpha and infiltration of neutrophils at the wound site of mice fed a diet containing <1 (deficient), 50 (control), 500, or 1000 microg zinc/g diet. Zinc deficiency reduced the rate of wound closure and mRNA levels of IL-1beta and TNF-alpha and attenuated infiltration of neutrophils at the wound site compared with controls. Interestingly, zinc supplementation at 1000 microg/g delayed the rate of wound closure and decreased mRNA levels of TNF-alpha and infiltration of neutrophils compared with mice fed the control diet. These findings demonstrate that zinc deficiency and high-dose zinc supplementation delay wound healing as a result of altered inflammatory responses and suggest that adequate zinc supplementation may have beneficial effects on the inflammatory responses to enhance cutaneous wound healing.

Stable Expression of Angiopoietin-1 and Other Markers by Cultured Pericytes: Phenotypic Similarities to a Subpopulation of Cells in Maturing Vessels During Later Stages of Angiogenesis In Vivo

Pericytes have been difficult cells to study because they do not maintain their characteristic phenotype in vitro, and they begin to express fibroblast markers after only a few days in culture. We now report methods for the isolation, purification, culture, and repurification of human dermal pericytes from mixed cell populations using an immunoaffinity-magnetic bead approach coupled with the 3G5 IgM monoclonal antibody that is specific for a pericyte surface ganglioside. These purified cells could be expanded in culture, and they maintained their pericyte phenotype for up to 8 days. In addition, they strongly expressed angiopoietin-1 (Ang-1) but not angiopoietin-2, Tie-1, or Tie-2; in contrast, dermal microvascular endothelial cells exhibited a reciprocal expression pattern. These findings are important because the close proximity of endothelial cells and pericytes has often made it difficult to determine with certainty the specific cell type(s) that expressed each of these proteins in situ. Extending our in vitro findings to two models of angiogenesis in vivo, we demonstrated a subpopulation of Ang-1–expressing cells that appeared in maturing microvessels during later stages of cutaneous wound healing and vascular permeability factor/vascular endothelial growth factor–induced angiogenesis. Our results provide strong evidence that Ang-1 is expressed by pericytes in vitro and in vivo and that the role proposed for Ang-1 in vessel maturation in development can be extended to vessel maturation after angiogenesis in adult tissues.