Rapid hemostasis by nanofibrs of polyhydroxyethyl methacrylate / polyglycerol sebacic acid: Fabrication, characterization, cytotoxicity and effectiveness on bleeding animal model

Rapid hemostasis by nanofibrs of polyhydroxyethyl methacrylate / polyglycerol sebacic acid: Fabrication, characterization, cytotoxicity and effectiveness on bleeding animal model
Jale varshosaz,¹ Zahra Choupannejad ,^2,* Mohsen minaiyan,³ Anoushe Zargar Kharazi,⁴
1. Novel Drug Delivery System Research Center, Department of Pharmaceutics, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Science, Isfahan,Iran
2. Novel Drug Delivery System Research Center, Department of Pharmaceutics, School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Science, Isfahan,Iran
3. Department of Pharmacology,School of Pharmacy and Pharmaceutical Sciences, Isfahan University of Medical Science, Isfahan,Iran
4. Department of Biomaterials, Tissue Engineering and Nanotechnology, School of Advanced Technologies in Medicine, Isfahan University of medical Science, Isfahan, Iran
Introduction: Uncontrollable hemorrhage is a primary reason for death in war, car accidents, and surgical procedures. Dehydration of blood causes half of the death in war. It is reported that more than 30% of deaths occur in 5-10 minutes as bleeding originates of hemorrhagic shock in pre-hospital emergency services. The fast bleeding control method does not only reduce death but also help to achieve optimal recovery. Bleeding control includes mechanical barriers or chemical inducing hemostatic pathways to help the natural blood coagulation process. Polyglycerol sebacic acis(PGS) is a biodegradable polymer, which is also widely used in tissue engineering and wound heling. This polymer is recommended for soft tissue engineering because it has elastomeric properties, biocompatibility, linear degradation rate, and nontoxic products. Polyhydroxyethyl methacrylate (PHEMA) is another biodegradable and biocompatible polymer used in tissue engineering. PHEMA has been used to prepare sponges, which can adhere to blood proteins and is also used to prepare spherical, porous particles containing ethamsylate and aminocaproic acid, the hemostatic drugs for embolization. Tranexamic acid (TA) is generally administered as capsule from oral route to control menorrhagia. It has a bitter taste and gastrointestinal side effects. Intravenous route is used in surgeries but it has thromboembolism risk and gets slow to therapeutic level. On the other hand TA has shown positive wound healing effect. Due to the low plasma level in topical use, direct topical use of TA in hemorrhage can reduce bleeding volume and bleeding time without systemic side effects. The purpose of the present study was designing nanofibers of PGS/PHEMA loaded with TA for homeostasis to reduce the severity and bleeding time.
Methods: Glycerol and sebacic acid were mixed with a molar stoichiometric ratio of 1:1 and stirred at 120 ˚C in a nitrogen atmosphere for 24h and pressure less than 50 mTor. Porous nanofibers of polyglycerol sebacate (PGS), polyhydroxyethyl methacrylate (PHEMA) and tranexamic acid (TA) were prepared by electrospinning method. The nanofibers were optimized for their morphology, diameter size, porosity, TA loading, release profile, hydrophilicity, water absorption and mechanical behavior. Their cytotoxicity was studied by MTT assay on L929 cells. The hemostasis control on tail cut model in rats was investigated.
Results: The best formulation contained 35% of the total polymers, 20% of PGS and 10% of TA to total polymers. These nanofibers had 64% porosity, 8.59% water sorption and 1.47% weight loss after 28 days with no cytotoxicity on L929 cells. TA loaded nanofibers showed significantly less bleeding volume than other groups, but no significant difference in bleeding time was seen with the blank nanofibers.
Conclusion: The nanofibers of PGS/PHEMA were electrospun and loaded with TA. The processing parameters were optimized to obtain nanofibers with a smooth, bead-less morphology and least diameter. The results demonstrated that the voltage and speed of injection of the solution significantly changed the nanofibers diameter. Increasing the total polymer content of the nanofibers increased thickness of nanofibers. PGS content and TA content of the nanofibers caused improvement of their mechanical behaviors and increasing the PHEMA content of the nanofibers caused more porous structure. By increasing the PGS content the mean pore area increased. The presence of PHEMA provided good hydrophilicity for absorbing the blood and slow biodegradability. The composite nanofibers were non-toxic on mouse fibroblast cells. Rapid drug release, which was completed within a few minutes provided rapid effectiveness of the nanofibers and decreased the bleeding time compared to the control groups. From the obtained results, it may be concluded that the designed dressing of TA may provide a prominent way for hemostasis compared to the conventional treatments of bleeding control, which reduces the risk of hemorrhagic shock by decreasing the volume and time of bleeding and induces faster recuperation of the injuries. The designed nanofibrous dressing may be useful in buccal, mucosal or battlefield and surgeries.
Keywords: hemostats, tranexamic acid, nanofiber, Polyglycerol sebacate , polyhydroxyethyl methacrylate ,