Semi-Solid Product of Medicinal Leech Enhances Wound Healing in Rats

Background: Wound causes a series of cascading reactions such as cellular damage, inflammatory reactions, and infection. Objectives: The present study aimed to prepare pharmaceutical skin product from lyophilized Hirudo orientalis leech source with active enzyme and proteins, having wound healing and anti-coagulation activities. Methods: The present study focused on the bioactive protein of Hirudo orientalis with anti-trypsin and anti-coagulation activity. First, biological active substance was prepared from lyophilized leech conducted on resultant 5% leech cream. Skin absorption of 5% leech cream was evaluated by Franz cell test, and in vivo wound healing capability of the leech cream was compared with phenytoin cream. In addition, the microbial control of final product was summed-up. Further, the quality control analysis and protein purification in final product were conducted using elution procedure on Hirudin protein-related region. Finally, the anticoagulant activity of the resultant protein was observed. Results: Using the result of the experiments investigating the successful effect containment function of the trypsin, we established the decreased rate of UV absorption in the area of 410 nm in question based on increases in the amount of cream. With 0, 100, 200 and 300 mg of 5% leech cream, absorbance (A) rate was: A (1.9, 1.47, 0.753 and 0.00) (P < 0.05; √2 = 21.6). In anticoagulation activity increased rate of partial thromboplastin time (PTT) (sec) (mean ± SEM), (26 ± 0.00, 34.22 ± 0.61 and 42 ± 0.71) with the 0, 100, and 200 mg final product was established. No bacterial colony growth was observed in the microbial culture analysis. In vivo results demonstrated that leech cream and phenytoin could promote wound regeneration by accelerating the re-epithelialization process and initial angiogenesis among all treatment groups. Conclusions: In the wound healing, we usually seek to reduce pain and inflammation, beside epitelialization and angiogenesis. Leech saliva contains biological substances that we have proven in this study. Our expectations for wound healing will be met with this pharmaceutical product citing the result of this study. The leech product proved to be the best selection for cosmetic with the added capabilities of renovation of the hair glands and regular skin form and typical depth of outer layer of skin. Thus, the leech cream can be considered as a better substitute for the conventional wound healing products.