Application of zein-based dural sealant in the repair of dural defects

English Keywords:Dural defect  Cerebrospinal fluid leakage  Zein  Dural sealant

English Abstract:

  【Abstract】 Objectives: To observe the biocompatibility of a novel zein-based dural sealant and verify its related biosafety and adhesive effect in aqueous environments, and observe its repair efficacy in rabbit dura mater and rat spinal dura mater defects. Methods: A novel zein-based dural sealant was fabricated, then lyophilized into patch form or partially ground for application. The biocompatibility of zein-based dural sealant was verified at the cellular level via cell counting Kit-8(CCK-8) assay, live/dead staining assay on PC12 cells, and hemolysis assay on rabbit red blood cells. Its absorbable properties were investigated through subcutaneous degradation experiments. The burst pressure of the zein-based dural sealant was validated using a modified porcine spinal dura mater in vitro model and a rabbit cranial dura mater in vivo model following ASTM F2392 standard, with a parallel comparison to fibrin glue. For the rabbit dural defect repair model, successfully modeled Japanese white rabbits were randomly divided into three groups based on treatment: blank group, fibrin glue group, and zein-based sealant group. Postoperatively, hematoxylin-eosin(HE) staining was used to observe the growth and repair of new fibers at the dural defect site; MRI examination was performed to assess CSF leakage sealing; And enzyme-linked immunosorbent assay(ELISA) was employed to evaluate the impact of sealing efficacy on the expression of inflammatory factors(including COX-2 and TNF-α) in CSF. The same grouping strategy was adopted in the rat spinal dura mater defect model to observe CSF leakage sealing effects across groups. Results: The zein-based dural sealant was successfully prepared. CCK-8 assay results showed that cell viability remained above 80% across all concentrations and time points; Hemolysis rate was below 5%; And Calcein/PI staining indicated a viable cell rate exceeding 90%. In vivo degradation experiments demonstrated complete resorption of the sealant within 7-10 days, with no significant inflammatory response observed via HE staining. The burst pressure of the zein-based sealant was approximately 92.2±12.3cmH2O, which was significantly higher than that of fibrin glue(36.0±3.2cmH2O) under the same conditions. In the rabbit dural defect model, HE staining revealed faster and denser ingrowth of new fibrous tissue in the zein-based sealant group compared with the blank and fibrin glue groups. MRI results confirmed more reliable CSF leakage sealing in the zein-based sealant group, accompanied by lower expression levels of inflammatory factors(COX-2 and TNF-α) in CSF. In the rat spinal dura mater defect model, gross observation also verified superior CSF leakage sealing performance of the zein-based sealant relative to the other two groups. Conclusions: The zein-based dural sealant exhibits excellent biocompatibility. Its outstanding adhesive performance in aqueous environments enables more effective sealing of CSF leaks, and it provides a favorable physiological microenvironment for fibroblasts, thereby achieving superior repair of dural defects.