Migration Of Human Fibroblasts Research Articles

Neutrophil extracellular traps with low concentrations induce proliferation and migration of human fibroblasts via activating CCDC25/ILK/PI3K/AKT pathway

Neutrophil plays an important role in the early stage of wound healing. Excessive neutrophil extracellular traps (NETs) which are produced by neutrophil have been proved as a significant unfavorable factor for wound healing by impairing the function of human skin fibroblasts. However, the effect of NETs with low concentrations on fibroblasts remains unclear. The interaction between CCDC25 and ILK in fibroblast were found to be activated by NET-DNA with low concentrations. Additionally, phosphorylation of PI3K and AKT was increased in NET-stimulated fibroblasts but was inhibited by CCDC25 knockout or ILK knockdown. NETs-induces proliferation and migration ability of fibroblast were also reduced by application of CCDC25 sgRNAs or ILK shRNAs. This study demonstrates that low concentrations of NETs, upon biding with CCDC25, activates the ILK/PI3K/AKT signaling pathway in skin fibroblasts, leading to increased cell proliferation and migration activity.

AT2R Activation Improves Wound Healing in a Preclinical Mouse Model.

Abnormal skin healing resulting in chronic wounds or hypertrophic scarring remains a major healthcare burden. Here, the antifibrotic angiotensin II type 2 receptor (AT2R) signaling pathway was modulated to determine its impact on cutaneous wound healing. Balb/c mice received two splinted full-thickness wounds. Topical treatments with the selective AT2R agonist compound 21 (C21) and/or selective antagonist PD123319 or saline vehicle were administered until sacrifice on post-wounding days 7 or 10. The rate of wound re-epithelialization was accelerated by PD123319 and combination treatments. In vitro, C21 significantly reduced human fibroblast migration. C21 increased both collagen and vascular densities at days 7 and 10 post-wounding and collagen I:III ratio at day 10, while PD123319 and combination treatments decreased them. Genes associated with regeneration and repair were upregulated by C21, while PD123319 treatment increased the expression of genes associated with inflammation and immune cell chemotaxis. C21 treatment reduced wound total leukocyte and neutrophil staining densities, while PD123319 increased these and macrophage densities. Overall, AT2R activation with C21 yields wounds that mature more quickly with structural, cellular, and gene expression profiles more closely approximating unwounded skin. These findings support AT2R signal modulation as a potential therapeutic target to improve skin quality during wound healing.

The concerted action of SEPT9 and EPLIN modulates the adhesion and migration of human fibroblasts.

Septins are cytoskeletal proteins that participate in cell adhesion, migration, and polarity establishment. The septin subunit SEPT9 directly interacts with the single LIM domain of epithelial protein lost in neoplasm (EPLIN), an actin-bundling protein. Using a human SEPT9 KO fibroblast cell line, we show that cell adhesion and migration are regulated by the interplay between both proteins. The low motility of SEPT9-depleted cells could be partly rescued by increased levels of EPLIN. The normal organization of actin-related filopodia and stress fibers was directly dependent on the expression level of SEPT9 and EPLIN. Increased levels of SEPT9 and EPLIN enhanced the size of focal adhesions in cell protrusions, correlating with stabilization of actin bundles. Conversely, decreased levels had the opposite effect. Our work thus establishes the interaction between SEPT9 and EPLIN as an important link between the septin and the actin cytoskeleton, influencing cell adhesion, motility, and migration.

Proanthocyanidins from Vaccinium vitis-idaea L. Leaves: Perspectives in Wound Healing and Designing for Topical Delivery.

The compositions and health-beneficial properties of lingonberry leaves (Vaccinium vitis-idaea L.) are well established; however, their proanthocyanidins are still heavily underutilized. Optimizing their delivery systems is key to enabling their wider applications. The present study investigates the phytochemical and ‘wound-healing’ properties of proanthocyanidin-rich fraction(s) (PRF) from lingonberry leaves as well as the development of optimal dermal film as a proanthocyanidin delivery system. The obtained PRF was subjected to HPLC-PDA and DMAC analyses to confirm the qualitative and quantitative profiles of different polymerization-degree proanthocyanidins. A ‘wound healing’ in vitro assay was performed to assess the ability of PRF to modulate the wound environment for better healing. Low concentrations of lingonberry proanthocyanidins were found to accelerate ‘wound‘ closures, while high levels inhibited human fibroblast migration. Fifteen dermal films containing PRF were prepared and evaluated based on their polymer (MC, HEC, PEG 400) compositions, and physical, mechanical, and biopharmaceutical properties using an experimental design. The composition containing 0.30 g of MC, 0.05 g of HEC, and 3.0 g of PEG 400 was selected as a promising formulation for PRF delivery and a potentially effective functional wound dressing material, supporting the need for further investigations.

Free-cell therapeutics and mechanism of exosomes from adipose-derived stem cells in promoting wound healing: current understanding and future applications.

Free-cell therapeutics and mechanism of exosomes from adipose-derived stem cells in promoting wound healing: current understanding and future applications.

Antibacterial composite hydrogels of graphene quantum dots and bacterial cellulose accelerate wound healing.

The increased antibiotic resistance of pathogenic bacteria requires intense research of new wound healing agents. Novel wound dressings should be designed to provide wound disinfection, good moisture, and fast epithelization. In this study, bacterial cellulose (BC) was impregnated with graphene quantum dots (GQDs) for potential use in wound healing treatment. The BC was successfully loaded with approximately 11.7wt% of GQDs. The actual release of GQDs from new designed composite hydrogels were 13%. Novel GQDs-BC hydrogel composites are biocompatible and showed significant inhibition towards Staphylococcus aureus and Streptococcus agalactiae and bactericidal effect towards Methicillin-resistant Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. The in vitro healing analysis showed significant migration of human fibroblasts after the GQDs-BC hydrogels application. Furthermore, after 72 h exposure to GQDs-BC, endothelial nitric oxide synthase, vascular endothelial growth factor A, matrix metallopeptidase 9, and Vimentin gene expression in fibroblast were significantly upregulated promoting angiogenesis. GQDs-BC hydrogel composites showed very good wound fluid absorption and water retention, which satisfies good dressing properties. All obtained results propose new designed GQDs-BC hydrogels as potential wound dressings.

Tremella fuciformis Inhibits Melanogenesis in B16F10 Cells and Promotes Migration of Human Fibroblasts and Keratinocytes.

Natural skin whiteners have been investigated for centuries. The development of preparations that safely achieve whitening of hyper-pigmented skin lesions is a challenge for the cosmetics industry. Furthermore, promoting rapid wound healing and minimizing inflammation in injured skin are key to prevent from abnormal pigmentation in scar tissue. Natural products, including the fungus Tremella fuciformis (TF), are attracting attention as potential sources of lead compounds for these applications. We investigated the in vitro effects of TF on melanogenesis in murine B16F10 cells. Melanin and tyrosinase levels were measured after treatment with TF. Wound healing in human keratinocytes (HaCaT) and fibroblasts (Detroit 551) was also determined via cell migration assay prior to TF exposure. TF significantly decreased melanin content and tyrosinase expression in a concentration-dependent manner in B16F10 cells. Furthermore, TF promoted wound healing in human HaCaT keratinocytes and Detroit 551 fibroblasts. TF proved effectively on inhibiting melanogenesis and promoting wound healing in vitro, demonstrating its potential as a novel skin-whitening agent. However, further clinical studies of safety and efficacy are required.

Cytotoxicity and Effect of Topical Application of Tranexamic Acid on Human Fibroblast in Spine Surgery

In spinal surgery, considerable blood loss is increasingly treated with the local application of tranexamic acid (TXA). However, little is known about its cytotoxicity and effect on human fibroblasts. This study was to identify the effect of TXA solution on human fibroblast at different concentrations and exposure times invitro. To mimic the actual clinical situation, human fibroblasts were subjected to both limited and chronic exposure to various clinically relevant concentrations of TXA to mimic different ways of topical administration. At time points after treatment, the viability, proliferation, apoptosis, collagen synthesis, adhesion, and migration of fibroblasts were analyzed invitro. Limited exposure (10 minutes) to a high concentration of TXA (100 mg/mL) did not affect the viability, proliferation, and apoptosis of fibroblasts, and chronic exposure to low concentration of TXA (≤12.5 mg/mL) exerted little effect on viability, proliferation, apoptosis, collagen synthesis, adhesion, and migration of human fibroblasts (P > 0.05). However, the chronic exposure to a high concentration of TXA (≥25 mg/mL) can inhibit the viability, proliferation, collagen synthesis, adhesion and migration, and induce apoptosis of fibroblasts. Although limited exposure to high concentration of TXA and chronic exposure to low concentration of TXA exerted little effect on fibroblasts, chronic exposure to high concentration of TXA can lead to fibroblast injury.

Enzymatically triggered graphene oxide released from multifunctional carriers boosts anti-pathogenic properties for promising wound-healing applications

Spurred by recent progress in biomaterials and therapeutics, stimulus-responsive strategies that deliver an active substance in temporal-, spatial-, and dose-controlled fashions have become achievable. Implementation of such strategies necessitates the use of bio-safe materials that are sensitive to a specific pathological incitement or that, in response to a precise stimulus, undergo hydrolytic cleavage or a change in biomolecular conformation. An innovative design of polymeric stimulus-responsive systems should controllably release a drug or degrade the drug carrier in response to specific lesion enzymes. Wound healing is a great challenge due to various hidden factors such as pathogenic infections, neurovascular diseases, excessive exudates, lack of an effective therapeutic delivery system, low cell proliferation, and cell migration. In addition, long-term use of antibiotics in chronic wound management can result in side effects and antimicrobial resistance. Novel treatments with antibacterial pharmaceuticals thus vitally need to be developed. Recently, graphene and graphene family members have emerged as shining stars among biomaterials for wound-healing applications due to their excellent bioactive properties, which can overcome limitations of current wound dressings and fulfill wound-healing requirements. Herein, we developed a feasible approach to impregnate graphene oxide (GO) into genipin-crosslinked gelatin (3GO) hydrogels to enzymatically control GO release. The developed hydrogels were characterized by chemical, physical, morphological, and cellular analyses. The results proved that the 3GO1 hydrogel is biocompatible and significantly enhanced the mechanical strength by encapsulating GO. Moreover, the rate of GO release depended on the crosslinking degree and environmental enzyme levels. Enzymatically released GO displayed uniform dispersity, retained its antibacterial activities against Staphylococcus aureus and Pseudomonas aeruginosa through sharp edges and wrapping mechanisms, and promoted human fibroblast migration. This multifunctional hydrogel we developed with antibacterial efficacy is suitable for future application as wound dressings.

IL-1β Impaired Diabetic Wound Healing by Regulating MMP-2 and MMP-9 through the p38 Pathway.

Diabetes mellitus is one of the most prominent metabolic disorders in the world, and insulin resistance in diabetic patients leads to several complications including increased inflammation and delayed wound healing. Fibroblast migration and reepithelialization play a significant role in wound healing. In this study, we explored the effects of IL-1β signaling on proliferation and migration of human fibroblasts from diabetic wound tissues. We observed elevated levels of IL-1β in samples from diabetic patients when compared to normal wound tissues. At high concentrations, IL-1β inhibited cell proliferation and migration in ex vivo fibroblast cultures. Moreover, expression of matrix metalloproteinases (MMPs) was upregulated, and tissue inhibitor of metalloproteinases (TIMPs) was downregulated in diabetic wound tissues and cells. These effects were regulated by levels of IL-1β. Furthermore, IL-1β induced p38 phosphorylation thereby activating the p38 MAPK pathway that in turn regulated the expression of MMPs and TIMPs. Together, our study identifies a novel mechanism behind delayed wound closure in diabetes mellitus that involves IL-1β-dependent regulation of cell proliferation and migration.

Synthesis by gamma irradiation of hyaluronic acid-polyvinyl alcohol hydrogel for biomedical applications.

Hyaluronic acid (HA) is one of the most attractive natural polymers employed in biomaterials with biological applications. This polysaccharide is found in different tissues of the body because it is a natural component of the extracellular matrix; furthermore, it has crucial functions in cell growth, migration, and differentiation. Since its biological characteristics, HA has been utilized for the new biomaterial's development for tissue engineering, such as hydrogels. These hydrophilic macromolecular networks have gained significant attention due to their unique properties, making them potential candidates to be applied in biomedical fields. Different mechanisms to obtain hydrogels have been described. However, the research of new non-toxic methods has been growing in recent years. In this study, we prepared a new hydrogel of HA and polyvinyl alcohol by the cost-effective technique of cross-linking by gamma irradiation. The hydrogel was elaborated for the first time and was characterized by several methods such as Fourier Transform Infrared Spectroscopy, Differential Scanning Calorimetry, Thermogravimetric Analysis, and Scanning Electron Microscopy. Likewise, we evaluated the cytotoxicity of the biomaterial and its influence on cell migration in human fibroblasts. Furthermore, we provide preliminary evidence of the wound closure effect in a cellular wound model. The novel hydrogel offers an increase of HA stability with the potential to expand the useful life of HA in its different medical applications.

Restraint of Human Skin Fibroblast Motility, Migration, and Cell Surface Actin Dynamics, by Pannexin 1 and P2X7 Receptor Signaling.

Wound healing is a dynamic process required to maintain skin integrity and which relies on the precise migration of different cell types. A key molecule that regulates this process is ATP. However, the mechanisms involved in extracellular ATP management are poorly understood, particularly in the human dermis. Here, we explore the role, in human fibroblast migration during wound healing, of Pannexin 1 channels and their relationship with purinergic signals and in vivo cell surface filamentous actin dynamics. Using siRNA against Panx isoforms and different Panx1 channel inhibitors, we demonstrate in cultured human dermal fibroblasts that the absence or inhibition of Panx1 channels accelerates cell migration, increases single-cell motility, and promotes actin redistribution. These changes occur through a mechanism that involves the release of ATP to the extracellular space through a Panx1-dependent mechanism and the activation of the purinergic receptor P2X7. Together, these findings point to a pivotal role of Panx1 channels in skin fibroblast migration and suggest that these channels could be a useful pharmacological target to promote damaged skin healing.

Butter oil (ghee) enrichment with aromatic plants: Chemical characterization and effects on fibroblast migration in an in-vitro wound healing model

Gheeis a dairy product widely consumed in India, north-Africa, and Middle East countries, havingbeneficialpharmacologicaleffects. Thisstudyaimsto characterize the effects of aromatic plants addition (rosemary and clove) on the nutritional, volatile and oxidative profile of cow ghee and to evaluate the effect of flavored ghee on the fibroblasts migration during wound healing in vitro assay. Two flavored ghee products were obtained byadding clove (CG) and rosemary (RG) as aromatic plants through maceration incattletraditional ghee (BT). It was revealed that enriched ghee samples had significantly lower peroxide values (6.76 and 6.80 meqO2 /kg) compared to control samples (8.20 meqO2 /kg). Moreover, the addition of rosemary and clove change the volatile profile, and increased the retinol levels of ghee (BT: 1.3 mg/kg; CG: 1.9 mg/kg; and RG: 3.05 mg/kg). Liquid-chromatography analyses revealed the presence of targeted phenolic compounds such as carnosic acid, rutin and gallic acid in CG and RG, showing thus, the transfer of polyphenols from aromatic plants into the ghee matrix. On the other hand, the fatty acid composition of ghee remained unchanged. The major components of the prepared ghee samples contributed to rising significantly the human fibroblast migration in wound healing in vitro assay. The results obtained underline that the flavored ghee samples could improve skin regeneration, making them potentials therapeutic ingredients in skincare formulations.

In vitro antibacterial activity of extracts from Samoan medicinal plants and their effect on proliferation and migration of human fibroblasts

Ethnopharmacological relevanceThe prevalence of different types of chronic wounds, due to the ageing population and increase incidence of diseases, is becoming a worldwide problem. Various medicinal plants used in folk medicine have demonstrated wound healing and antimicrobial properties, and some of these species are currently used in commercial preparations. Despite the well-documented and rich tradition of the use of local herbs for the treatment of skin injuries in Samoan folk medicine, their wound healing potential has not yet been systematically studied. Aim of the studyInvestigation into the in vitro antibacterial activity of ethanol extracts from 14 medicinal plants used in Samoan traditional medicine for the healing of wounds, burns and sores, and their effects on the proliferation and migration of human fibroblasts. Materials and methodsThe antibacterial activity of these extracts was tested against pathogens associated with infected skin injuries, using the broth microdilution method. The effect on migration, proliferation and viability of human dermal fibroblasts was evaluated using wound healing scratch assay, cell proliferation assay, and thiazolyl blue tetrazolium bromide cytotoxicity test. ResultsThe extracts from Cerbera manghas, Commelina diffusa, Kleinhovia hospita, Mikania micrantha, Omalanthus nutans, Peperomia pellucida, Phymatosorus scolopendria, Piper graeffei, Psychotria insularum, and Schizostachyum glaucifolium inhibited the growth of Staphylococcus aureus at the minimum inhibitory concentration (MIC) of ≥4 μg/mL, whereas C. manghas and P. pellucida produced the same MIC against both Escherichia coli and Pseudomonas aeruginosa. Among the antibacterially active species, C. diffusa, K. hospita, P. scolopendria, P. insularum, and S. glaucifolium did not produce toxicity towards the standard line of normal adult human dermal fibroblasts (IC80 > 128 μg/mL). In addition, extracts from Barringtonia asiatica, C. manghas, M. micrantha, O. nutans, P. insularum, and Piper graeffei stimulated significant migration of dermal fibroblasts, while M. micrantha, O. nutans, and P. insularum did not affect cell proliferation at a concentration of 32 μg/mL. ConclusionsThe results suggest that the above-mentioned species of Samoan medicinal plants can be used for the development of new wound healing agents. However, further phytochemical and pharmacological research is needed regarding the isolation and identification of their active constituents.

Use of Human Umbilical Cord and Its Byproducts in Tissue Regeneration.

The fresh or cryopreserved human umbilical cord (HUC) and its byproducts, such as cells and extracts, have different uses in tissue regeneration. Defining what HUC byproduct is more effective in a particular application is a challenge. Furthermore, the methods of isolation, culture and preservation, may affect cell viability and regenerative properties. In this article, we review the HUC and its byproducts’ applications in research and clinical practice. We present our results of successful use of HUC as a patch to treat gastroschisis and its potential to be applied in other conditions. Our in vitro results show an increase in proliferation and migration of human fibroblasts by using an acellular HUC extract. Our goal is to promote standardization of procedures and point out that applications of HUC and its byproducts, as well as the resulting advances in regenerative medicine, will depend on rigorous quality control and on more research in this area.

Exosomes derived from human amniotic epithelial cells accelerate diabetic wound healing via PI3K-AKT-mTOR-mediated promotion in angiogenesis and fibroblast function.

BackgroundDiabetic wounds are one of the most common and serious complications of diabetes mellitus, characterized by the dysfunction of wound-healing-related cells in quantity and quality. Our previous studies revealed that human amniotic epithelial cells (hAECs) could promote diabetic wound healing by paracrine action. Interestingly, numerous studies demonstrated that exosomes derived from stem cells are the critical paracrine vehicles for stem cell therapy. However, whether exosomes derived from hAECs (hAECs-Exos) mediate the effects of hAECs on diabetic wound healing remains unclear. This study aimed to investigate the biological effects of hAECs-Exos on diabetic wound healing and preliminarily elucidate the underlying mechanism.MethodshAECs-Exos were isolated by ultracentrifugation and identified by transmission electron microscopy, dynamic light scattering and flow cytometry. A series of in vitro functional analyses were performed to assess the regulatory effects of hAECs-Exos on human fibroblasts (HFBs) and human umbilical vein endothelial cells (HUVECs) in a high-glycemic microenvironment. High-throughput sequencing and bioinformatics analyses were conducted to speculate the related mechanisms of actions of hAECs-Exos on HFBs and HUVECs. Subsequently, the role of the candidate signaling pathway of hAECs-Exos in regulating the function of HUVECs and HFBs, as well as in diabetic wound healing, was assessed.ResultshAECs-Exos presented a cup- or sphere-shaped morphology with a mean diameter of 105.89 ± 10.36 nm, were positive for CD63 and TSG101 and could be internalized by HFBs and HUVECs. After that, hAECs-Exos not only significantly promoted the proliferation and migration of HFBs, but also facilitated the angiogenic activity of HUVECs in vitro. High-throughput sequencing revealed enriched miRNAs of hAECs-Exos involved in wound healing. Kyoto Encyclopedia of Genes and Genomes and Gene Ontology analyses have shown that the target genes of the top 15 miRNAs were highly enriched in the PI3K-AKT pathway. Further functional studies demonstrated that the PI3K-AKT-mTOR pathway was necessary for the induced biological effects of hAECs-Exos on HFBs and HUVECs, as well as on wound healing, in diabetic mice.ConclusionsOur findings demonstrated that hAECs-Exos represent a promising, novel strategy for diabetic wound healing by promoting angiogenesis and fibroblast function via activation of the PI3K-AKT-mTOR pathway.

Nintedanib decreases muscle fibrosis and improves muscle function in a murine model of dystrophinopathy

Duchenne muscle dystrophy (DMD) is a genetic disorder characterized by progressive skeletal muscle weakness. Dystrophin deficiency induces instability of the sarcolemma during muscle contraction that leads to muscle necrosis and replacement of muscle by fibro-adipose tissue. Several therapies have been developed to counteract the fibrotic process. We report the effects of nintedanib, a tyrosine kinase inhibitor, in the mdx murine model of DMD. Nintedanib reduced proliferation and migration of human fibroblasts in vitro and decreased the expression of fibrotic genes such as COL1A1, COL3A1, FN1, TGFB1, and PDGFA. We treated seven mdx mice with 60 mg/kg/day nintedanib for 1 month. Electrophysiological studies showed an increase in the amplitude of the motor action potentials and an improvement of the morphology of motor unit potentials in the animals treated. Histological studies demonstrated a significant reduction of the fibrotic areas present in the skeletal muscles. Analysis of mRNA expression from muscles of treated mice showed a reduction in Col1a1, Col3a1, Tgfb1, and Pdgfa. Western blot showed a reduction in the expression of collagen I in skeletal muscles. In conclusion, nintedanib reduced the fibrotic process in a murine model of dystrophinopathy after 1 month of treatment, suggesting its potential use as a therapeutic drug in DMD patients.

Polyelectrolyte complex of Aloe vera, chitosan, and alginate produced fibroblast and lymphocyte viabilities and migration

Chitosan, sodium alginate and gel of Aloe vera (Aloe barbadensis Miller) were employed for the preparation of polyelectrolyte complexes at pH 4 and 6. FT-IR spectroscopy analysis showed evidence on complexes formation and incorporation of the Aloe vera gel. The ζ potential determination of the polyelectrolyte complexes revealed the presence of surface charges in the range of −20 to −24 mV, which results in stable systems. The dynamic moduli exhibited a high dependence on angular frequency, which is commonly found in solutions of macromolecules. The materials showed human fibroblast and lymphocyte viabilities up to 90% in agreement with null cytotoxicity. The polyelectrolyte complexes at pH 6 with Ca2+ were stable, showed high water absorption, satisfactory morphology, pore size and rigidity, characteristics that allowed significant human fibroblast migration in wound closure in vitro assays.

Podoplanin increases the migration of human fibroblasts and affects the endothelial cell network formation: A possible role for cancer-associated fibroblasts in breast cancer progression.

In our previous studies we showed that in breast cancer podoplanin-positive cancer-associated fibroblasts correlated positively with tumor size, grade of malignancy, lymph node metastasis, lymphovascular invasion and poor patients’ outcome. Therefore, the present study was undertaken to assess if podoplanin expressed by fibroblasts can affect malignancy-associated properties of breast cancer cells. Human fibroblastic cell lines (MSU1.1 and Hs 578Bst) overexpressing podoplanin and control fibroblasts were co-cultured with breast cancer MDA-MB-231 and MCF7 cells and the impact of podoplanin expressed by fibroblasts on migration and invasiveness of breast cancer cells were studied in vitro. Migratory and invasive properties of breast cancer cells were not affected by the presence of podoplanin on the surface of fibroblasts. However, ectopic expression of podoplanin highly increases the migration of MSU1.1 and Hs 578Bst fibroblasts. The present study also revealed for the first time, that podoplanin expression affects the formation of pseudo tubes by endothelial cells. When human HSkMEC cells were co-cultured with podoplanin-rich fibroblasts the endothelial cell capillary-like network was characterized by significantly lower numbers of nodes and meshes than in co-cultures of endothelial cells with podoplanin-negative fibroblasts. The question remains as to how our experimental data can be correlated with previous clinical data showing an association between the presence of podoplanin-positive cancer-associated fibroblasts and progression of breast cancer. Therefore, we propose that expression of podoplanin by fibroblasts facilitates their movement into the tumor stroma, which creates a favorable microenvironment for tumor progression by increasing the number of cancer-associated fibroblasts, which produce numerous factors affecting proliferation, survival and invasion of cancer cells. In accordance with this, the present study revealed for the first time, that such podoplanin-mediated effects can affect tube formation by endothelial cells and participate in their pathological properties in the tumor context. Our experimental data were supported by clinical studies. First, when IDC and DCIS were analyzed by immunohistochemistry according to the presence of podoplanin-expressing cells, the numbers of cancer-associated fibroblasts with high expression of this glycoprotein were significantly higher in IDC than in DCIS cases. Second, using immunofluorescence, the co-localization of PDPN-positive CAFs with blood vessels stained with antibody directed against CD34 was observed in tumor stroma of IDC samples.

Effects of conditioned medium from LL-37 treated adipose stem cells on human fibroblast migration.

Adipose stem cell-conditioned medium may promote human dermal fibroblast (HDF) proliferation and migration by activating paracrine peptides during the re-epithelization phase of wound healing. Human antimicrobial peptide LL-37 is upregulated in the skin epithelium as part of the normal response to injury. The effects of conditioned medium (CM) from LL-37 treated adipose stem cells (ASCs) on cutaneous wound healing, including the mediation of fibroblast migration, remain to be elucidated, therefore the aim of the present study was to determine how ASCs would react to an LL-37-rich microenvironment and if CM from LL-37 treated ASCs may influence the migration of HDFs. The present study conducted migration assays with HDFs treated with CM from LL-37 treated ASCs. Expression of CXC chemokine receptor 4 (CXCR4), which controls the recruitment of HDFs, was analyzed at the mRNA and protein levels. To further characterize the stimulatory effects of LL-37 on ASCs, the expression of stromal cell-derived factor-1α (SDF-1α), a CXC chemokine, was investigated. CM from LL-37-treated ASCs induced migration of HDFs in a time- and dose-dependent manner, with a maximum difference in migration observed 24 h following stimulation with LL-37 at a concentration of 10 µg/ml. The HDF migration and the expression of CXCR4 in fibroblasts was markedly increased upon treatment with CM from LL-37-treated ASCs compared with CM from untreated ASCs. SDF-1α expression was markedly increased in CM from LL-37 treated ASCs. It was additionally observed that SDF-1α blockade significantly reduced HDF migration. These findings suggest the feasibility of CM from LL-37-treated ASCs as a potential therapeutic for human dermal fibroblast migration.