Polydopamine-Coated Polycaprolactone Electrospun Nanofiber Membrane Loaded with Thrombin for Wound Hemostasis

Abstract

Hemorrhagic shock is the primary cause of death in patients with severe trauma, and the development of rapid and efficient hemostatic methods is of great significance in saving the lives of trauma patients. In this study, a polycaprolactone (PCL) nanofiber membrane was prepared by electrospinning. A PCL-PDA loading system was developed by modifying the surface of polydopamine (PDA), using inspiration from mussel adhesion protein, and the efficient and stable loading of thrombin (TB) was realized to ensure the bioactivity of TB. The new thrombin loading system overcomes the disadvantages of harsh storage conditions, poor strength, and ease of falling off, and it can use thrombin to start a rapid coagulation cascade reaction, which has the characteristics of fast hemostasis, good biocompatibility, high safety, and a wide range of hemostasis. The physicochemical properties and biocompatibility of the PCL-PDA-TB membrane were verified by scanning electron microscopy, the cell proliferation test, the cell adhesion test, and the extract cytotoxicity test. Red blood cell adhesion, platelet adhesion, dynamic coagulation time, and animal models all verified the coagulation effect of the PCL-PDA-TB membrane. Therefore, the PCL-PDA-TB membrane has great potential in wound hemostasis applications, and should be widely used in various traumatic hemostatic scenarios.

Keywords: electrospinning; hemostatic method; polydopamine modification; thrombin; trauma.

Figure 1

(A) SEM and EDS elemental mapping images of PCL and PCL–PDA fibrous membranes. (B) Fiber diameter distribution of PCL and PCL–PDA fibrous scaffolds (n = 200). (C) Water contact angles of PCL and PCL–PDA fibrous membranes (n = 3, ** p < 0.01).

Figure 2

(A) CLSM images of FITC-BSA loaded onto PCL and PCL–PDA fibrous membranes. (B) Fluorescence intensity of FITC-BAS on PCL and PCL–PDA fibrous membranes (n = 6, ** p < 0.01).

Figure 3

(A) Cell viability of PCL and PCL–PDA fibrous membranes incubated with Fibroblasts cells for 1 day. (B) SEM images of Fibroblasts cells grown on PCL and PCL–PDA fibrous membranes. (C) Representative CLSM images of fibroblasts cells after 1 day, 3 days, and 5 days of culture on PCL and PCL–PDA fibrous membranes.

Figure 4

(A) Photographs of the wounds that were embedded in the PCL, PCL–PDA, control group and blank group samples immediately following surgery (day 0), and for 3, 7, and 14 days. Histological analysis of subcutaneous tissue of the wounds after embedding surgery at 14 days. Photographs and histological analysis for rat internal organs (heart, lung, stomach, liver, spleen, and kidney) after 14 days embedding experiment with PCL–PDA (B) and blank group (C), respectively.

Figure 5

(A) Photographs from the in vitro blood-clotting measurement and the blood clotting index of the supernatant absorbance for PCL, PCL–PDA, and PCL–PDA–TB (n = 3, * p < 0.05, versus PCL group; ** p < 0.01, versus PCL group; *** p < 0.001, versus PCL group; # p < 0.05, versus PCL–PDA group; ## p < 0.01, versus PCL–PDA group). The SEM images of blood cells (B) and activated platelets (C) on the PCL, PCL–PDA, and PCL–PDA–TB surface (blue arrow).

Figure 6

(A) Schematic diagram of the rat femoral artery hemostasis model. (B) Blood loss and (C) hemostatic time on the rat femoral artery hemostasis model by using control, PCL, PCL–PDA, PCL–PDA–TB and commercial. (D) Images of the hemostasis (from left to right) by use of control, PCL, PCL–PDA, PCL–PDA–TB, and commercial (n = 6, *** p < 0.001).

Figure 7

(A) Schematic diagram of the rat liver homeostasis model. (B) Blood loss and (C) hemostatic time on the rat liver homeostasis model by using control, PCL, PCL–PDA, PCL–PDA–TB and commercial. (D) Images of the hemostasis (from left to right) by use of control, PCL, PCL–PDA, PCL–PDA–TB and commercial (n = 6, *** p < 0.001).