Of Preparation Of Earthworm Protein Gels And For The Treatment Of Wound

In this study, one earthworm protein was extracted and purified from Eisenia Foelida. As gel raw material, Earthworm protein was a protein substances, its stability was affected by outside influence easily. This project developed a gel including solid portion and liquid portion separately. The earthworm protein and gel matrix were saved as solid form. When using, the liquid portion was poured into the solid portion, then the gel swelled in no more than2minutes.The research about this gel focused on the effect of promoting wound healing and anti-inflammation. Moreover, correlative mechanism of earthworm protein gel was researched. Main content and conclusions are as follows:1. Studying on Earthworm protein characteristics and stability. Main characteristics of three batches of Earthworm protein, including the yield, pH of different density Earthworm protein solution, specific rotations, protein content, polysaccharides content, biological activity, were almost the same. The results demonstrated procedures of extracting and purifying earthworm was robust, and the main characteristics were reproducible.Stressing testing indicated:three batches of earthworm protein appearance, weight difference, difference in protein content, biological activity had no significant difference, to comply with the requirements. The stability of earthworm protein characters provided a good basis for preparation of earthworm protein gel.2. Preparation and properties study of the earthworm protein gel. The gel matrix screened in the preformulation study was carbopol940, which was disposed as follows:the concentration of carbopol0.13%-0.2%was diluted with water, swelled after overnight, saved24hours at-20℃, freezed with freeze dryer. With addition of a certain amount of triethanolamine, the new Carbopol powder swelled within2minutes. Electron microscope scanning tested the spatial structure of carpobol changed, which may be the main reason that swelling speed of carbopol be faster. By screening of response surface design, the formulation of earthworm protein gel was:earthworm protein powder (drug)0.1g, carbomer940disposed (gel matrix)0.3g, triethanolamine (pH regulator)3.72g, glycerin (moisturizer)12.5g, ethylparaben (preservative)0.01g, mannitol (protein freeze-dried protecting agent) 0.0001g, distilled water (solvent)84mL. Earthworm protein gel properties were performed, such as rheology and preliminary stability test.3. The earthworm protein gel was performed to circular excision model and linear incision model. Morphological, biochemical and histopathologic indices showed as follows:earthworm protein gel with high dose group can improve the wound hydroxyproline, myeloperoxidase, malondialdehyde content to increase collagen, fibroblasts, capillary numbers and reduce free radical attack; meanwhile, it can reduce the wound inflammatory factor TNF-α levels, increase inflammation reaction factor IL-10levels to improve the inflammation, then promote wound healing, shorten wound healing time.In this study, one protein, which can promote wound healing, was extracted and separated from the earthworm. Compared with fresh earthworm, this protein has advantages in storage, transportation and so on. The earthworm protein gel was prepared successfully and applied to rat wound model. The method preparation of this gel provided the basis for gel of pharmaceutical production for its high reproducibility and stability. Moreover, this study provided some evidences for further study of main ingredients developed into burn preparation for external use and extends the earthworm medicinal field.