Bioinspired Non-Immunogenic Multifunctional Sealant for Efficient Blood Clotting and Suture-Free Wound Closure

Abstract

Engineering bioinspired peptide-based molecular medicine is an emerging paradigm for the management of traumatic coagulopathies and inherent bleeding disorder. A hemostat-based strategy in managing uncontrolled bleeding is limited due to the lack of adequate efficacy and clinical noncompliance. In this study, we report an engineered adhesive peptide-based hybrid regenerative medicine, sealant 5, which is designed integrating the structural and functional features of fibrin and mussel foot-pad protein. AFM studies have revealed that sealant 5 (55.8 ± 6.8 nN adhesive force) has higher adhesive force than fibrin (46.4 ± 7.3 nN adhesive force). SEM data confirms that sealant 5 retains its network-like morphology both at 37 and 60 °C, inferring its thermal stability. Both sealant 5 and fibrin exhibit biodegradability in the presence of trypsin, and sealant 5 also showed biocompatibility in the presence of fibroblast cells. Engineered sealant 5 efficiently promotes hemostasis with enhanced adhesiveness and less blood-loss than fibrin. In vivo data suggests that in heparinized conditions, sealant 5 ceases bleeding at 212.3 ± 15.1 s, whereas fibrin halts bleeding at 294.3 ± 21.4 s and blood-loss is ∼4-fold less in sealant 5 than in fibrin. In a heparinized system, sealant 5 facilitates faster blood-clotting than fibrin (∼82 s faster) and RADA-16, a reported peptide-based sealant (∼113 s faster). Additionally, in the case of sealant 5, the process of clotting mimicry-like fibrin is independent of the body's own coagulation system. Sealant 5 efficiently halts bleeding for both external and internal wounds, even for a heparinized system overcoming the bacterial infection. ELISA data and PMBC cell proliferation data support the nonimmunogenic feature of sealant 5. Though fibrin and sealant 5 have exhibited comparable efficacy in suture-free wound closure, in vivo H&E staining images have revealed infiltration of very few immune cells as well as the presence of abundant collagen formation in the case of sealant 5-treated wound. Such nature-inspired non-immunogenic sealants offer exciting possibilities for the treatment of uncontrolled bleeding vis-a-vis wound closure.

Cohesion Design-Led Tough Sealants with Controllably Dissolvable Properties

“…Altogether, these data proved that SEAgel can not only efficiently stopped the bleeding but also may prevent the liver adhesion after surgery. To further prove the safety of the SEAgel, immunogenic response experiments were carried out following the previous report . The ELISA results showed that a very low IgG response was observed for SEAgel, which was not significantly different when compared to that of the commercially available Fibrin glue (Figure g).…”

“…To further prove the safety of the SEAgel, immunogenic response experiments were carried out following the previous report. 38 The ELISA results showed that a very low IgG response was observed for SEAgel, which was not significantly different when compared to that of the commercially available Fibrin glue (Figure 5g). The pro-inflammatory cytokine IFN-γ was also detected showing that SEAgel did not induce the over-expression of the cytokine, further demonstrating the good biocompatibility of SEAgel (Figure 5f).…”

“…This experiment was carried out following previous reports . Interferon-gamma (IFN-γ) mRNA expression level was quantified by real-time polymerase chain reaction (PCR).…”

“…The enzyme-linked immunosorbent assay (ELISA) experiments were carried out following previous reports with minor modifications. 38 First, SD rats (100−140 g) were immunized with SEAgel, SEgel, and Fibrin glue by applying them into the liver incisions. 7 days post-surgery, 2 mL of blood was collected from each rat.…”

“…This experiment was carried out following previous reports. 38 Interferon-gamma (IFN-γ) mRNA expression level was quantified by real-time polymerase chain reaction (PCR). SD rats with incisions were treated with PBS, SEgel, SEAgel, and Fibrin glue.…”