Extraction And Evaluation The Hemostatic Performance Of Spongin From Spongia Officinalis

Sponge is the most primitive multicellular animal.During the evolution of 600 million years,sponges evolved a unique collagen-like spongin.Spongin is a protein complex with stable structure,high cross-linking and multi-fibrous monomer polymerization.It has strong acid resistance,high temperature resistance and enzymatic hydrolysis resistance.Studies have shown that spongin has excellent biocompatibility,rich pore structure and acid corrosion resistance.It has potential application in the field of biomaterials such as tissue repair scaffolds and bioelectrodes.Based on the structural characteristics of spongin,this paper attempts to explore the feasibility of this material as a hemostatic dressing.This study talks about the research of spongin extraction and hemostratic performance evaluation,and it is expected that more applications of this material can be found.In this study,Bath sponge was used as raw material to extract spongin.Through the Sirius red staining and the conteng tracking of hydroxyproline,the following extraction process was determined:firstly,the impurities were separated by hydrochloric acid and sodium hydroxide to obtain the water-insoluble spongin skeleton(A).The composition has a yield of 60.41%and a hydroxyproline content of 3.36%.The component A was trypsinized to obtain a spongin material with a yield of 36%and a hydroxyproline content of 5.67%.Subsequently,the skeleton was subjected to freeze-grinding treatment(70 Hz/s,15 min,-10 ?)to obtain two materials with significant difference in particle size,one of which was flocculent,accounting for 30%,and its saturated water absorption was 4050%.The rate is 75.40%.The fibers are connected under the electron microscope and the mesh is uniform.The material is numbered as SX;the second is powder granules,accounting for 70%,the saturated water absorlption rate is 994%,the porosity is 90.46%,and each electron microscope The individual monomers are dendritic and the material is numbered SFM.The component A is dissolved with hydrogen peroxide,dialyzed and freeze-dried to obtain a water-soluble material.The saturated water absorption rate of the material is 362%,the porosity is 86.67%,and it is in the form of a sheet under an electron microscope,and is numbered SR.Fourier infrared absorption spectroscopy analysis showed that the triple helix structure of SX and SFM materials remained intact,while the triple helix structure of SR material might unwind.From the physical parameters such as water absorption,porosity and micro-bifurcation,the hemostatic effects of these three spongin materials may be significantly different.Next,we experimented with in vitro coagulation experiments and animal experiments of three spongin materials.,and Yunnanbaiyao drug or type I collagen was used as a positive control.The results of in vitro coagulation experiments showed that the mean clotting time of Yunnanbaiyao drug,type I collagen,SX,SFM and SR were 151 s?167 s?235 s?118 s?194 s,and the blank group was 222 s,indicating that the hemostatic performance of SFM was significantly better than other groups;The results of the blood loss of the rat tail tail showed that the blood loss of rats treated with type I collagen,SX,SFM and SR hemostasis were 22.8 mg,22.3 mg,3.6 mg and 28.1 mg,respectively,compared with the blank group(93.1 mg).There was a significant decrease,especially in the SFM group;the hemostasis time showed that the hemostasis time of type I collagen,SX,SFM,and SR hemostasis was 714 s,371 s,129 s,and 697 s,respectively,while the blank group was after 900 s.Still no coagulation,these results indicate that all three spongins contribute to wound hemostasis,with SFM being particularly significant.This paper then described a preliminary study on the mechanism of hemostasis.In vitro red blood cell adhesion experiments showed that spongin materials have the function of inducing cell pseudopod migration and enrichment of cells;platelet aggregation experiments show that spongin materials have platelet aggregation and activation,providing thrombus support sites;platelet blood coagulation experiments show SX and SFM The material has the ability to cause blood clotting of the defective blood;the defibrinated blood coagulation experiment shows that the SFM and SR materials have the ability to cause blood clotting of the defective blood.The above results indicate that the coagulation mechanism of the spongin material itself can adsorb red blood cells and platelets,and its rich dendritic bifurcation promotes fibrin cross-linking and thrombus formation.The mechanism by which SFM and SR materials can cause blood clotting in blood cells with clotting factors remains to be explored.About the reason of SFM materials have better hemostatic effect than the other two spongin materials,we speculate that the material particles can fully contact the wound,and its rich dendritic bifurcation can promote thrombus cross-linking,so it can quickly penetrate the entire wound.Scope to build a cross-linked thrombus and quickly stop bleeding.In order to be applied to medical hemostasis,this study conducted biocompatibility analysis of SFM and SR materials.The results showed that these two materials did not have antibacterial activity,but they did not promote the growth of bacteria;they met the national standard in hemolysis rate,and there was no potential stimuli on the skin.The acute systemic toxicity test of rats confirmed that they had no acute body,toxicity.Based on the experimental results above,spongin materials,especially SFM materials,have effective procoagulant properties,special hemostasis ability to overcome clotting factor defects,and good biocompatibility,and have wide application value and development prospects in the field of hemostasis.Subsequent research will focus on SFM materials to further optimize the hemostatic function and preparation process of SFM materials.