Chronic venous disease – Part II: Proteolytic biomarkers in wound healing

Abstract

Venous leg ulcers (VLU) are characterized by sustained proteolytic microenvironment impairing the healing process. Wound fluid (WF) reflect the biomolecular activities occurring within the wound area; however, it is unclear if WF from different healing phases have different proteolytic profiles and how VLU microenvironment affects the wound healing mechanisms. We investigated the proteolytic network of WF from distinct VLU phases, and in WF- and LPS-stimulated THP-1 monocytes treated with glycosaminoglycan sulodexide, a well known therapeutic approach for VLU healing. WF were collected from patients with VLU during inflammatory (Infl) and granulating (Gran) phases. WF and THP-1 supernatants were analyzed for nine matrix metalloproteinases (MMP) and four tissue inhibitors of metalloproteinases (TIMP) by multiplex immunoassays. Our results demonstrated that: 1) WF from Infl VLU contained significantly increased concentrations of MMP-2, MMP-9, MMP-12, TIMP-1, and TIMP-2 compared to Gran WF; 2) WF from Gran VLU showed significantly increased levels of MMP-1, MMP-7, MMP-13, and TIMP-4 compared to Infl WF; 3) LPS- and WF-stimulation of THP-1 cells significantly increased the expression of several MMP compared to untreated cells; 4) Sulodexide treatment of both LPS- and WF-stimulated THP-1 significantly down-regulated the release of several MMPs. Our study provides evidence-based medicine during treatment of patients with VLU. WF from Infl and Gran VLU have different MMP and TIMP signatures, consistent with their clinical state. The modulation of proteolytic pathways in wound microenvironment by glycosaminoglycan sulodexide, provide insights for translating research into clinical practice during VLU therapy.

Sulodexide for the Symptoms and Signs of Chronic Venous Disease: A Systematic Review and Meta-analysis

“…On the basis of sulodexide's antiplatelet [38], anti-inflammatory [11,39,40] and anti-proteolytic [41] effects, as well as the protective effect on the glycocalyx layer [42], it may interfere with the pathogenesis of both primary and secondary disease in patients with CVD [43,44]. In fact, sulodexide has been shown to have favourable effects on the most distressing clinical sign of CVD, venous ulcers, of different origins [45,46].…”

“…We excluded 41 studies that were originally considered as potentially eligible. The reasons for exclusion were: no clinical data reported (n = 13) [12,13,29,[39][40][41][73][74][75][76][77][78][79], indication other than CVD with data on CVD symptoms not available/not extractable (n = 13) [80][81][82][83][84][85][86][87][88][89][90][91][92], no extractable data on symptoms (n = 9) [36,[93][94][95][96][97][98][99][100][101], preliminary partial publication (n = 3) [102][103][104], data reported but not stratified by treatment (n = 1) [105], review (n = 1) [47] or duplicate (n = 1) [98].…”

Sulodexide inhibits angiogenesis via decreasing Dll4 and Notch1 expression in mouse proepicardial explant cultures

“…There is evidence that SDX protects endothelial lining and vessel wall in the microcirculation mostly due to glycocalyx restoration and inhibition of leukocyte adhesion to ECs. Additionally, SDX suppresses inflammatory process in vitro and in vivo, downregulating expression, release, and activity of proinflammatory cytokines such as IL-1b, IL-6, IL-8, TGF-b1, and TNF-a, as well as metalloproteinases, both from ECs and macrophages in vivo and in vitro, as was observed in LPS activated human monocytic cell line THP1 [12,[15][16][17].…”

Efeitos in vitro da radiação infravermelha longa em fibroblastos humanos oriundos do leito de feridas crônicas de etiologia venosa

“…MMP-21, MMP-23)72,73,74 . Em comum, todas essas metaloproteinases possuem um conjunto de características que as tornam únicas entre as enzimas; são elas: presença de um íon de zinco no sítio catalítico, secreção pela célula forma inativa (zimógeno), sequência de aminoácidos grosseiramente semelhantes, especificidade para degradar pelo menos um componente da matriz e inibição pelas TIMPs70 .…”

“…Apesar dessa degradação proteolítica ser parte essencial do reparo e remodelação durante a cicatrização, a função das MMPs não está restrita ao "turn over" da matriz71,77 . Elas também estão envolvidas direta ou indiretamente, em outras tarefas: 1) remoção do tecido desvitalizado, 2) migração e proliferação celular, por degradar proteínas da matriz responsáveis pelas ligações célula-célula, célula-MEC, 3) interações epidérmica-mesenquimal durante a migração de queratinócitos, 4) angiogênse, 5) regulação da atividade de certos fatores de crescimento/citocina (libertação, ativação, silenciamento), 6) alteração de receptores da superfície celular71,72,78 .Em tecidos normais, a expressão de MMPs, quando ocorre, acontece em níveis baixos, mas sua expressão e ativação são rapidamente induzidas quando a remodelação ativa do tecido é necessária. Nessa circunstância, a expressão de MMPs pode ser induzida em muitos tipos celulares(queratinócitos, fibroblastos, células endoteliais e células inflamatórias, como linfócitos, monócitos e macrófagos) em resposta a citocinas, hormônios, oncogenes e contato das células com a MEC ou outros tipos celulares.…”