Carbonized Cellulose Aerogel Derived from Waste Pomelo Peel for Rapid Hemostasis of Trauma-Induced Bleeding

Abstract

Uncontrollable massive bleeding caused by trauma will cause the patient to lose a large amount of blood and drop body temperature quickly, resulting in hemorrhagic shock. This study aims to develop a hemostatic product for hemorrhage management. In this study, waste pomelo peel as raw material is chosen. It underwent processes of carbonization, purification, and freeze-drying. The obtained carbonized pomelo peel (CPP) is hydrophilic and exhibits a porous structure (nearly 80% porosity). The water/blood absorption ratio is significantly faster than the commercial Gelfoam and has a similar water/blood absorption capacity. In addition, the CPP showed a water-triggered shape-recoverable ability. Moreover, the CPP shows ideal cytocompatibility and blood compatibility in vitro and favorable tissue compatibility after long terms of subcutaneous implantation. Furthermore, CPP can absorb red blood cells and fibrin. It also can absorb platelets and activate platelets, and it is capable of achieving rapid hemostasis on the rat tail amputation and hepatectomized hemorrhage model. In addition, the CPP not only can quickly stop bleeding in the rat liver-perforation and rabbit heart uncontrolled hemorrhage models, but also promotes rat liver and rabbit heart tissue regeneration in situ. These results suggest the CPP has shown great potential for managing uncontrolled hemorrhage.

Keywords: biocompatibility; carbonized pomelo peel; hemostasis; uncontrollable massive bleeding.

Figure 1

The physical characterization of carbonized pomelo peel sponge (CPP). A) SEM images show the internal structure of CPP after hydration thermal reaction at 140 °C, 160 °C, and 180 °C for 8, 10, and 12 h, respectively. B) Porosity quantification of different types of CPP, the gelfoam (Gel) as a positive control. C) The compression strain‐stress curve of different CPPs that carbonized at 140 °C for 8, 10, and 12 h, respectively. D) The compression strain‐stress curve of different CPPs that carbonized at 160 °C for 8, 10, and 12 h, respectively. E) The compression strain‐stress curve of different CPPs that carbonized at 180 °C for 8, 10, and 12 h, respectively. F) The compression strain‐stress curve of different CPPs that carbonized at 140 °C, 160 °C, and 180 °C for 12 h, respectively. G) The compression strain‐stress curve of (160 °C, 8 h) CPP, (160 °C, 10 h) CPP, and (180 °C, 10 h) CPP under wet conditions. H) The surface contact angle of (160°C, 8 h) CPP, (160 °C, 10 h) CPP, (180 °C, 10 h) CPP, and gelfoam. I) Quantification of the contact angle of different CPPs and gelfoam. **p <0.01, ***p <0.001, ****p <0.0001.

Figure 2

The swelling and shape recoverable behaviors of CPP. A) Photographs visualize the hydration‐induced shape recovery of various CPPs and gelfoam before and after compression in water or blood. B) The water absorption rate of various CPPs and gelfoam over time. C) The maximum water absorption capacity of various CPPs and gelfoam. D) The blood absorption rate of various CPPs and gelfoam over time. E) The maximum blood absorption capacity of various CPPs and gelfoam. *p <0.05, **p <0.01, ***p <0.001, ****p <0.0001.

Figure 3

In vitro biocompatibility of CPP. A) Live/Dead staining of L929 cells treated with different CPP extracts for 1 and 3 days. B) The proliferation of L929 cells co‐cultured with different CPP extracts for 1, 3, and 7 days. C) In vitro hemolysis test of different CPPs and gelfoam with 625, 1250, and 2500 µg concentrations, respectively. D) Quantitative analysis of hemolysis test of different CPPs and gelfoam.

Figure 4

The tissue compatibility of different CPPs. H&E staining images of different CPPs and gelfoam after subcutaneous implantation for 7, 14, 28, and 56 days. CT: connective tissue, CPP: carbonized pomelo peel.

Figure 5

The in vitro hemostatic ability of CPPs. A) Hemoglobin binding capacity of various CPPs, gauze, and gelfoam at 1, 2, 3, 4, and 5 min. B) Blood clotting index (BCI) (%) of different CPPs, gauze, and gelfoam across detection time points. C) SEM images exhibiting RBC absorption on the surface of different CPPs, gauze, and gelfoam. D) Percentage of adhered RBCs on different CPPs, gauze, and gelfoam surfaces. E) SEM visualization of fibrin adhesion on different CPPs, gauze, and gelfoam surfaces. F) SEM images showing platelet adhesion on different CPPs, gauze, and gelfoam surfaces. G) Zeta potential of the different CPPs and gelfoam sponge. H) Activated partial thromboplastin time (APTT) analysis of the different CPPs and gelfoam. *p <0.05, ***p <0.001, ****p <0.0001.

Figure 6

The performance of the carbonized CPPs on the rat tail amputation and hepatectomized hemorrhage model. A) Photographs showing the hemostatic ability of different CPPs, gauze, and gelfoam on the rat tail amputation hemorrhage model. B,C) Quantitative analysis of the total blood loss and hemostatic time of different CPPs, gauze, and gelfoam on the rat tail amputation hemorrhage model. D) Photographs showing the hemostatic ability of different CPPs, gauze, and gelfoam on the rat hepatectomized hemorrhage model. E,F) Quantitative analysis of the total blood loss and hemostatic time of different CPPs, gauze, and gelfoam on the rat hepatectomized hemorrhage model. *p <0.05, **p <0.01, ***p <0.001, ****p <0.0001.

Figure 7

The hemostasis and tissue repair ability of CPPs on the rat liver‐perforation hemorrhage model. A) Photographs showing the hemostatic ability of different CPPs, gauze, and gelfoam on the rat liver‐perforation hemorrhage model. B,C) Quantitative analysis of the total blood loss and hemostatic time of different CPPs, gauze, and gelfoam on the rat liver‐perforation hemorrhage model. D) H&E staining images exhibit the interface between liver and CPP, gauze, and gelfoam after 7 and 14 days of treatment.

Figure 8

The hemostasis and tissue repair ability of CPPs on the rabbit heart uncontrolled hemorrhage model. A) Photographs showing the hemostatic ability of different CPPs, gauze, and gelfoam on the rabbit heart uncontrolled hemorrhage model. B,C) Quantitative analysis of the total blood loss and hemostatic time of different CPPs, gauze, and gelfoam on the rabbit heart uncontrolled hemorrhage model. D) H&E staining images exhibit the interface between heart and CPP, gauze, and gelfoam after 7 and 14 days of treatment.