# Isolation, Characterization and Biological Evaluation of Collagen from Rhizostoma pulmo Jellyfish from the Sea of Azov for Biomedical Applications

## TL;DR

This paper explores collagen from Azov Sea jellyfish as a safe and sustainable material for biomedical uses like tissue engineering and wound healing.

## Contribution

The study introduces Rhizostoma pulmo jellyfish collagen as a novel marine source with potential for biomedical applications.

## Key findings

- Extracted collagen showed high yield (~26.2%) and a type I-like profile with α-, β-, and γ-components.
- Collagen sponges supported 3D cell growth and tumor-cell dissemination in an in vitro breast carcinoma model.
- Low endotoxin levels (0.461 EU/µL) and no cytotoxicity were observed in tested conditions.

## Abstract

Collagen is a major extracellular-matrix protein widely used in regenerative medicine, yet conventional terrestrial sources raise biosafety and acceptability concerns, motivating the search for marine alternatives. This study evaluates the jellyfish Rhizostoma pulmo (R. pulmo) from the Azov Sea as a sustainable collagen source and assesses its suitability for biomedical materials. Acid-soluble collagen was extracted using 0.5 M acetic acid and purified by salt precipitation and dialysis, followed by physicochemical/structural characterization (sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE), Limulus amebocyte lysate (LAL) endotoxin testing, transmission electron microscopy (TEM), and immunofluorescence with type I collagen antibodies) and biological evaluation in vitro (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cytotoxicity on MRC5 fibroblasts; adhesion and proliferation assays on HeLa cells). The extracted collagen showed a high yield (~26.2%), a type I-like electrophoretic profile with α-, β-, and γ-components, fibrillar ultrastructure by TEM, and positive type I collagen immunoreactivity; endotoxin levels were low (0.461 EU/µL), and no cytotoxicity was detected under the tested conditions. Porous collagen sponges/scaffolds were fabricated by lyophilization, displaying interconnected pores with an average size of ~80 µm and pH-dependent swelling, and they supported 3D cell growth and tumor-cell dissemination in an in vitro breast carcinoma scaffold model. Overall, Azov Sea R. pulmo collagen demonstrates promising structural quality, low endotoxin burden, and cytocompatibility, supporting its potential as a marine biomaterial for sponge/scaffold-based tissue engineering and wound-related applications.