Cytokine Loaded Biopolymers as a Novel Strategy to Study Stem Cells during Wound‐Healing Processes

Grimaldi, Annalisa; Banfi, Serena; Vizioli, Jacopo; Tettamanti, Gianluca; Noonan, Douglas M.; Eguileor, Magda de

doi:10.1002/mabi.201000452

Cited by 13 publications

(9 citation statements)

References 55 publications

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“…2d). As previously reported, following LPS stimulation the vasocentral and vasofibrous cells dissociated from each other [35, 36], and the vasofibrous cells gave rise to type I granulocytes [37]. Indeed, 30 min after LPS injection, several vasofibrous tissue cells crossing the thick muscle layers were readily recognizable by light microscopy due to their dark pigmentation (Fig.…”

Section: Resultssupporting

confidence: 62%

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AIF-1 and RNASET2 Play Complementary Roles in the Innate Immune Response of Medicinal Leech

et al. 2018

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Recent studies demonstrated that allograft inflammatory factor-1 (AIF-1) and RNASET2 act as chemoattractants for macrophages and modulate the inflammatory processes in both vertebrates and invertebrates. The expression of these proteins significantly increases after bacterial infection; however, the mechanisms by which they regulate the innate immune response are still poorly defined. Here, we evaluate the effect of bacterial lipopolysaccharide injection on the expression pattern of these genes and the interrelation between them during innate immune response in the medicinal leech, an invertebrate model with a simple anatomy and a marked similarity with vertebrates in inflammatory processes. Collectively, prokaryotic-eukaryotic co-cultures and in vivo infection assays suggest that RNASET2 and AIF-1 play a crucial role in orchestrating a functional cross-talk between granulocytes and macrophages in leeches, resulting in the activation of an effective response against pathogen infection. RNASET2, firstly released by granulocytes, likely plays an early antibacterial role. Subsequently, AIF-1⁺ RNASET2-recruited macrophages further recruit other macrophages to potentiate the antibacterial inflammatory response. These experimental data are in keeping with the notion of RNASET2 acting as an alarmin-like molecule whose role is to locally transmit a “danger” signal (such as a bacterial infection) to the innate immune system in order to trigger an appropriate host response.

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Section: Resultssupporting

confidence: 62%

“…2e). By transmission electron microscopy (TEM) analysis, vasofibrous cell-derived type I granulocytes [37] were clearly recognizable underneath the epithelium and characterized by small round electron-dense granules (Fig. 2f).…”

Section: Resultsmentioning

confidence: 99%

AIF-1 and RNASET2 Play Complementary Roles in the Innate Immune Response of Medicinal Leech

et al. 2018

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“…100 μl sterilized PBS containing 20 ng rHmAIF-1 functionally to characterize cells migrating under the influence of this factor. The best concentration of rHmAIF-1 required to induce significant cell migration was determined based on our previous work on leeches Grimaldi et al 2011). We tested three different concentrations (5 ng, 10 ng and 20 ng) and obtained the best response with the highest amount.…”

Section: Animals and Treatmentsmentioning

confidence: 99%

Homolog of allograft inflammatory factor-1 induces macrophage migration during innate immune response in leech

et al. 2014

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Allograft inflammatory factor-1 (AIF-1) is a 17-kDa cytokine-inducible calcium-binding protein that, in vertebrates, plays an important role in the allograft immune response. Its expression is mostly limited to the monocyte/macrophage lineage. Until recently, AIF-1 was assumed to be a novel molecule involved in inflammatory responses. To clarify this aspect, we have investigated the expression of AIF-1 after bacterial challenge and its potential role in regulating the innate immune response in an invertebrate model, the medicinal leech (Hirudo medicinalis). Analysis of an expressed sequence tag library from the central nervous system of Hirudo revealed the presence of the gene Hmaif-1/alias Hmiba1, showing high homology with vertebrate aif-1. Immunohistochemistry with an anti-HmAIF-1 polyclonal antibody revealed the constitutive presence of this protein in spread CD68(+) macrophage-like cells. A few hours after pathogen (bacterial) injection into the body wall, the amount of these immunopositive cells co-expressing HmAIF-1 and the common leucocyte marker CD45 increased at the injected site. Moreover, the recombinant protein HmAIF-1 induced massive angiogenesis and was a potent chemoattractant for macrophages. Following rHmAIF-1 stimulation, macrophage-like cells co-expressed the macrophage marker CD68 and the surface glycoprotein CD45, which, in vertebrates, seems to have a role in the integrin-mediated adhesion of macrophages and in the regulation of the functional responsiveness of cells to chemoattractants. CD45 is therefore probably involved in leech macrophage-like cell activation and migration towards an inflammation site. We have also examined its potential effect on HmAIF-1-induced signalling.

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“…The proliferation assay on MG sponges demonstrates that our consolidated experimental approach for macrophage isolation 18 , 33 , 34 does not cause damages to cells, which retain their proliferative ability. On the other hand, after MWCNT in vitro treatment this ability significantly decreases.…”

Section: Discussionmentioning

confidence: 82%

“…However, many of the in vitro studies, using cancer cell lines, do not take into account the typical intrinsic resistance to apoptosis of cancer cells 36 . Our original and innovative assay based on the use of the biomatrix matrigel (MG) to obtain in vitro expansion of leech macrophages 18 , 33 , 34 , 42 , which are also implicated in response against abiotic particles such as MWCNTs, is an important tool that can be used as quick sensitive model for aquatic pollution bio-monitoring. Thus, in living organisms and culture cells, measurement of ROS intracellular levels and cell proliferation and viability might provide a reliable method to quickly identify oxidative stress states induced by the presence of MWCNTs in the environment.…”

Section: Discussionmentioning

confidence: 99%

Cellular responses induced by multi-walled carbon nanotubes: in vivo and in vitro studies on the medicinal leech macrophages

Girardello

Baranzini

Tettamanti

et al. 2017

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The core characteristics of multi-wall carbon nanotubes (MWCNTs) are impressive and attractive for technology however, since their production and use is steadily increasing, their environmental dispersion could be potentially hazardous to animal and human health. For this reason, the identification of new methods and of reliable models to better understand MWCNT effects is essential. Here we propose the medicinal leech as an alternative model to assess the effects of MWCNTs on immune system. Our previous studies have already demonstrated that in vivo MWCNT treatment induces the activation of leech’s macrophages. Here we will focus on the direct effects of MWCNTs on these cells by isolating and culturing leech's macrophages by means of the consolidated Matrigel technique, followed by MWCNT in vitro treatment. Our results indicate that MWCNT administration causes both the decrease of cell proliferation rate and the increase of the apoptotic rate. Furthermore, since oxidative stress is linked with inflammation, reactive oxygen species has been evaluated confirming that their production rate increases after MWCNT treatment. Our experimental approaches demonstrate the ability of MWCNTs inducing a powerful inflammatory response and confirm that the medicinal leech is a good alternative model to study the possible harmful effects of any nanomaterial.

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Cytokine Loaded Biopolymers as a Novel Strategy to Study Stem Cells during Wound‐Healing Processes

Cited by 13 publications

References 55 publications

AIF-1 and RNASET2 Play Complementary Roles in the Innate Immune Response of Medicinal Leech

AIF-1 and RNASET2 Play Complementary Roles in the Innate Immune Response of Medicinal Leech

Homolog of allograft inflammatory factor-1 induces macrophage migration during innate immune response in leech

Cellular responses induced by multi-walled carbon nanotubes: in vivo and in vitro studies on the medicinal leech macrophages

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