Plasminogen Activator Inhibitor-1 Controls Bone Marrow-Derived Cells Therapeutic Effect Through MMP9 Signaling: Role in Physiological and Pathological Wound Healing

Abstract

We assessed the role of plasminogen activator inhibitor-1 (PAI-1) and matrix metalloproteinase 9 (MMP9) in wound healing process and in the bone marrow mononuclear cells (BMMNC)-related effects on physiological and pathological wound healing. A full thickness excision wound was created by removal of the skin on the midback of irradiated and nonirradiated animals. Angiogenesis and re-epithelialization were markedly increased in PAI-12/2 mice compared to wild-type (WT) animals. We revealed high MMP activity in tissue of PAI-12/2 animals. Of interest, the wound healing process was reduced in PAI-12/2:MMP92/2 animals compared to PAI-12/2 mice, suggesting a key role of MMP9 in beneficial effect of PAI-1 deficiency on wound closure. To unravel the role of PAI-1 in BMMNC relative effects, mice were treated with or without local injection of BMMNC isolated from WT, PAI-12/2, and PAI-12/2: MMP92/2 animals for 14 days (10 6 cells, n 5 6 per group). In WT nonirradiated mice, transplantation of BMMNC isolated from PAI-12/2 animals enhanced wound formation when compared with WT BMMNC. BMMNC differentiation into cells with endothelial phenotype was enhanced by PAI-1 deficiency. These effects were abrogated in PAI-12/2:MMP92/2 and MMP92/2 BMMNC. In addition, using chimeric mice, we demonstrated that PAI-1 deficiency environment increased the BMMNC-GFP recruitment to the wound site, whereas this effect was abrogated when using PAI-12/2:MMP92/2 BMMNC. PAI-1 deficiency, at least through MMP9 upregulation, enhanced wound healing and BMMNC therapeutic potential in irradiated and nonirradiated animals.

The mRNA expressions and immunohistochemistry of factors involved in angiogenesis and lymphangiogenesis in the early stage of rat skin incision wounds

“…These factors have been reported to be involved in various types of angiogenesis [10,13,[16][17][18][29][30][31], and may be involved in the skin wound healing in our system.…”

“…MMP9 is considered to play a role in angiogenesis through MMP-mediated release of biologically relevant cytokines from the matrix and cell surfaces [57]. MMP9 upregulation enhanced wound healing in mice under PAI1 deficiency [29]. On the other hand, re-epithelialization was reported to be delayed in the skin wound in MMP9 knockout mice [58].…”

“…(CSF3/G-CSF) [11][12][13][14][15], chemokine (C-X-C motif) ligand2 (CXCL2) [16,17], chemokine (C-X-C motif) ligand 12 (CXCL12/SDF1) [18][19][20][21], endothelin 1 (ET1) [22], fibroblast growth factor 1 (FGF 1) [23], hepatocyte growth factor (HGF) [24,25], hypoxia inducible factor 1 alpha (HIF1a) [26], leptin [27], matrix metallopeptidase 9 (MMP9) [18,28,29], serpine/plasminogen activator inhibitor 1 (PAI1) [29][30][31], platelet-derived growth factor-A (PDGF-A) [18,32], transforming growth factor alpha (TGFa) [33,34], transforming growth factor beta 1 (TGFb1) [35,36], tenomodulin (TNMD) [37], and troponin I type 2 (TNNI2) [38,39]. Factors known to be involved in lymphangiogenesis such as fibroblast growth factor 2 (FGF 2) [40,41], c-fos induced growth factor (FIGF/VEGF-D) [42,43], forkhead box C 2 (FOXC2) [44][45][46], and prospero homeobox 1 (PROX1) [46][47][48][49] are also included in the study.…”

4G/4G PAI-1 gene variant in a patient with non-healing ulcers

“…Single nucleotide insertion or deletions at the PAI-1 promoter sequence create PAI-1 gene polymorphisms with varying phenotypes. [1,5] For example, the insertion/ deletion located 675 bp upstream of the transcription site in the promoter region of the PAI-1 gene: a five guanine nucleotide tract is the 5G allele, whereas a deletion of one G nucleotide results in the 4G variant allele. [6] The homozygous 4G allele mutation results in PAI-1 overexpression and therefore elevated levels of PAI-1 in plasma.…”

“…[6] The homozygous 4G allele mutation results in PAI-1 overexpression and therefore elevated levels of PAI-1 in plasma. [1,3,5] Figure 4. t-PA (or u-PA) catalyzes the conversion of plasminogen to plasmin. Free t-PA can bind to plasminogen on the fibrin clot surface for increased stability.…”