AiP1, a protein from the coelomic fluid of sea star Astropecten indicus promotes wound healing and fibrinogenolysis in vitro

Abstract

Background: Marine animals are suspected to hold keys to counter several life-threatening diseases. Despite its vast biodiversity, the marine ecosystem is relatively unexplored for its therapeutic potential. This study was particularly aimed at finding a potent molecule against thrombosis from the coelomic fluid of a sea star (SCF). SCF is a reservoir of immunocytes, coelomocytes, cytokines, and other substances responsible for the exceptional regenerative potential and other bioactivities. Death due to thrombosis has increased significantly in the recent years. Healing of wounds caused due to accidents or surgical procedures is also of great significance, particularly for diabetic patients. Therefore, search for good thrombolytic and wound healing agents is being pursued by many research groups globally. Methods: SCF was fractionated using ion-exchange chromatography and size exclusion HPLC to yield a potent protein, AiP1. Proteolytic assays were performed using SDS-PAGE and zymography. Cell culture studies were performed as per standard protocols. Results: Purified protein, AiP1, showed fibrinogenolytic, fibrinolytic, and anti-platelet activities in a dose-dependent manner. Proteolytic activities of AiP1 were inhibited post treatment with EDTA, indicating its metalloproteinase nature. It was also devoid of cytotoxic, hemolytic, and phospholipase activities. Scratch wounds created on surface of HEK293 and HaCaT cells showed faster wound closure rates post treatment with AiP1 compared to untreated cells. Peptide mass fingerprinting showed the presence of vWFA-like domain in the central β sheet which may be responsible for its bioactivities. AiP1 was thus isolated and characterized for its thrombolytic and wound healing potential.Conclusion: Coelomic fluid of sea stars contains many bioactive compounds. One such compound is the protein AiP1 containing a vWFA-like domain. This protein showed significant wound healing and thrombolytic activities in vitro.

In vitro Toxicity Evaluation of Sterol Isolate from Echinoderm Stellaster equestris against Human Peripheral Blood

“…The results hence indicated that the crude coelomic fluid from the sea star Astropecten indicus is a safe entrant for drug discovery. 14 However the results obtained from the present study for the in vitro analysis of hemolytic assay correlates with the finding that the compound isolated sea star Stellaster equestris even in a dose dependent manner were non-hemolytic and seems to be safe. The present study revealed that the purified sample did not show any cytotoxic effect on the exposed lymphocytes even in the dose dependant manner on comparison with the positive control.…”

“…Previous study on the cell viability of the novel protein isolated from the coelomic fluid of sea stars Astropecten indicus reported to show increased cell viability in the time dependent manner indicating the protein molecule isolated from the coelomic fluid was not toxic. 14 Hence the present study correlates with the cell viability assay by trypan blue exclusion assay reporting to be non-toxic.…”

Seven-Arms Starfish “Luidia maculata” Characteristics of Coelomocytes and Promising GSK3-β Protein for Wound Healing: Cellular and Chemical Analyses

“…The preparation of the starfish coelomic fluid (CF) was performed by applying the same procedure described in the study of Baveja et al (2018Baveja et al ( , 2019; additional lengthy details were published by Abdel-Ghaffar and Youssef (2022). After centrifugation of the coelomic fluid, a pellet of cells was formed and prepared for microscopical examination.…”

Collective Locomotion of Human Cells, Wound Healing and Their Control by Extracts and Isolated Compounds from Marine Invertebrates

“…It is a benthic echinoderm that is typically found in the sediments of the intertidal and subtidal regions, where it feeds using infaunal molluscs, and represents a prototypic marine organism to study the effects of environmental pollution [169,170]. Recently, Baveja et al [171] isolated the AiP1 metalloproteinase from the coelomic fluid of A. indicus and demonstrated its fibrinolytic potential that is likely mediated by the von Willebrand factor (vWf)-like domain located in its central β sheet. Moreover, the ability of AiP1 to modulate the motile attitude of HaCaT keratinocytes and HEK293 embryonic kidney epithelial cells was checked through scratch wound healing assays demonstrating the additional AiP1-promoting effect on cell migration since vWf is known to sustain cell locomotory behavior.…”