Preparation And Properties Of Hemostatic Polysacch-based Hydrogel Loaded With Antitumor Active Ingredients

Background: As a kind of disease which is easy to relapse and difficult to cure,malignant tumor is a serious threat to human health.Nowadays,the treatment methods are diversified,but surgery still plays an important role in the comprehensive treatment.Due to the fragile general condition of patients after surgery,systemic therapy should be used after an interval of convalescence,resulting in a blank period of treatment,but making the remaining tumor cells get a chance to breathe.At this time,an appropriate strategy is needed to fill the gap in tumor treatment and achieve the purpose of continuous treatment.The hemostatic polysaccharide based hydrogel system prepared with sodium hyaluronate and chitosan derivatives as the matrix can be loaded with anti-tumor active ingredients for the use in radical tumor surgery,which is expected to become a reliable way to achieve the function of slow release anti-tumor drugs in situ and instant wound hemostasis,and to solve the clinical dilemma.Purpose: In order to deal with the adverse reactions caused by the application of non-degradable materials in vivo and the defects of the simple function of hemostatic materials,this paper constructed a hydrogel loaded with anti-tumor active ingredients of hemostatic hyaluronic acid and chitosan derivatives as the matrix for the local anti-tumor treatment and filling irregular wound bleeding in the operation of breast cancer and other malignant tumors.In order to develop multiple integrated biodegradable materials to fill the vacancy of clinical application.Methods: Hydrogels,a 3D cross-linked hydrophilic polymer with a physical structure similar to that of natural extracellular matrix(ECM),have attracted worldwide attention in the field of pharmaceutical materials,such as drug delivery.N-carboxyethyl chitosan(CEC)was synthesized by Michael addition reaction from sodium hyaluronate and chitosan with good biocompatibility,and then Na IO4 was used to modify the HA molecular chain to have active aldehyde group(A-HA).N-carboxyethyl chitosan compound hyaluronic acid(CEC/A-HA)hydrogels were prepared by Schiff base reaction in CEC and A-HA solution under physiological conditions.The physicochemical properties,cytotoxicity,hematotoxicity,biosafety,in vitro and in vivo inhibition of tumor cell growth and proliferation,and in vivo hemostatic performance of the polysaccharide hydrogel were evaluated.It is proposed that this system can not only be used to stop irregular wound immediately during the operation,but also retain the slow-release drugs implanted in the body to kill the possible residual tumor cells and reduce the risk of recurrence and metastasis.Results:1.CEC/A-HA hydrogel was successfully prepared by Michael addition,sodium periodate oxidation and other modification methods,and evaluated by various methods,the in vitro and in vivo degradation time,swelling ratio,water content,pore morphology,pore size and other aspects were in line with the conditions of hemostasis and drug loading and slow release in vivo degradable carrier;2.CEC/A-HA hydrogel material and its components were co-cultured with Rat rat fibroblast cells and observed by CCK8 method and dead/alive fluorescence staining for72 h respectively to evaluate the cytotoxicity of the material.The results showed that the cells showed good proliferation rate and vigorous growth characteristics.The CEC/A-HA hydrogel material was implanted subcutively into the back of SD rats by injection,and no acute,subacute or chronic toxicity was observed in the experimental animals during the6-month observation period.The hydrogel materials prepared in this study showed excellent biosafety characteristics through the evaluation of in vivo and in vitro experiments,which is an important prerequisite for clinical use in vivo.3.Both in vivo and in vitro experiments confirmed that,compared with free 5-FU,CEC/A-HA&5-FU achieved A stronger inhibitory effect on the growth and proliferation of tumor cells.In vitro,CEC/A-HA hydrogel loaded with 5-fluorouracil was co-cultured with MDA-MB-231 breast cancer cells.The results showed that the hydrogel could slow release 5-FU into the co-culture system and significantly inhibit the proliferation of MDA-MB-231 breast cancer cells.In vivo,a mouse 4T1 in situ breast cancer model was constructed.Peritumoral injection of CEC/A-HA&5-FU hydrogel was performed to observe the changes in tumor volume.The growth rate of tumor in the experimental group slowed down,which enhanced the anti-tumor effect of the drug in vivo.4.We constructed liver surface bleeding model and tail severed bleeding model of SD rats,and used CEC/A-HA hydrogel prepared in this study for hemostasis experiment.The results showed that the experimental group showed the characteristics of adaptive filling of liver tear wounds,and the hemostasis performance of this material was defined by the evaluation of hemostasis time and bleeding amount.