Fig 1.
SDS polyacrylamide gel electrophoresis pattern of collagens from jellyfish mesoglea.
STD: molecular weight marker; RTC: rat tail type I collagen; PSC: pepsin soluble collagen; ASC: acid soluble collagen.
Table 1.
Amino acid composition of the collagen from jellyfish (PSC) and calf-skin collagen (results are expressed as residues/1000 residues).
Table 2.
Cross-linking degree of jellyfish collagen sponges.
Fig 2.
Fourier transform infrared spectrum of different collagen sponges.
A, uncross-linked collagen sponge; B, collagen sponge cross-linked with 50mM EDC for 12h; C, collagen sponge cross-linked with 100mM EDC for 24h (The unit of the numbers in figure is cm-1).
Fig 3.
SEM images of cross-linked and uncross-linked lyophilized collagen sponges of different concentration.
A, B: cross-section and surface of 3.3 mg/ml collagen sponges cross-linked by 100mM EDC for 24 h, respectively. C, D: cross-section and surface of 10.0 mg/ml collagen sponges cross-linked by 100mM EDC for 24 h, respectively. E, F: cross-section and surface of 10.0 mg/ml collagen sponges uncross-linked, respectively.
Fig 4.
Water absorption capacity (WAC) of different sponges.
(A) shows the WAC of different collagen sponges (3.3 mg/ml and 2.5 mg/ml), and medical gauze group was set as control. The symbol (*) denotes statistical significance from control group (P < 0.05). (B) shows the sponges with different cross-linking degree fabricated by 3.3 mg/ml collagen have different WAC. Degree of cross-linking is expressed as EDC concentration (mM)/cross-linking time (h) (50/12, 50/24, 100/12 and 100/24 in this study). The symbol (*) and (**) denotes statistical significance from uncross-linked groups and 100/12 groups, respectively (P < 0.05).
Fig 5.
Comparison of weight loss of uncross-linked collagen (Control group) and cross-linked collagen groups.
Degree of cross-linking are expressed as EDC concentration (mM)/cross-linking time (h) (50/12, 50/24, 100/12 and 100/24 in this study). The symbol (*) denotes statistical significance from control group (P < 0.05).
Fig 6.
Viability of cells cultured in direct contact with various biomaterials at 1, 3 and 7 days, as determined by MTT assay.
Viability is expressed as a percentage of live cells compared to positive control cells: (1) uncross-linked 2.5 mg/ml collagen sponges; (2) 2.5 mg/ml collagen sponges cross-linked with 100 mM EDC for 24h; (3) uncross-linked 3.3 mg/ml collagen sponges; (4) 3.3 mg/ml collagen sponges cross-linked with 100 mM EDC for 24h.
Fig 7.
SEM images of blood cells and platelets adhered on the surface of collagen sponges.
(A) Aggregation and morphology of RBCs on the collagen sponge. (B) RBCs generated spiny pseudopodia (arrow). (C) Platelet adhered on the collagen sponge exhibited regular resting shape (arrow).
Fig 8.
Collagen sponges effect on platelets adhesion.
The adherent platelets on different sponges were observed by calcein-AM staining, and images were showed in (A) control group, (B) uncross-linked collagen sponges and (C) cross-linked collagen sponges. The numbers of platelets were counted (D). Scale bar, 30 μm.
Fig 9.
Results of whole blood clotting experiment.
The clotting formation capacity is expressed as blood clotting index (BCI), which reflect the free RBCs not trapped in clots. Lower BCI value means better clotting formation capacity. Control: medical gauze; (1) uncross-linked 2.5 mg/ml collagen sponges; (2) 2.5 mg/ml collagen sponges cross-linked with 100 mM EDC for 24h; (3) uncross-linked 3.3 mg/ml collagen sponges; (4) 3.3 mg/ml collagen sponges cross-linked with 100 mM EDC for 24h. The symbol (*) denotes statistical significance from control group (P < 0.05).
Fig 10.
Hemostatic effect of collagen sponges in a murine tail amputation model.
(A) Blood loss after tail amputation was conducted in 5 min. The symbol (*) denotes statistical significance from control group (P < 0.05). (B) Coagulation time of tail amputation experiment. The symbol (*) denotes statistical significance from blank (P < 0.05). The symbol (**) denotes statistical significance from control (P < 0.05). In the picture, (1) uncross-linked 2.5 mg/ml collagen sponges; (2) 2.5 mg/ml collagen sponges cross-linked with 100 mM EDC for 24h; (3) uncross-linked 3.3 mg/ml collagen sponges; (4) 3.3 mg/ml collagen sponges cross-linked with 100 mM EDC for 24h.