Nghiên cứu độc tính và hiệu quả điều trị vết thương phần mềm của cao lỏng LT tt tiếng anh

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MINISTRY OF EDUCATION AND TRAINING MINISTRY OF DEFENSE MILITARY INSTITUTE OF TRADITIONAL MEDICINE TRUONG MINH TUAN STUDY THE TOXICITY AND EFFICIENCY OF LT EXTRACT ON WOUND Speciality: Traditional medicine Code: 62720201 DOCTOR THESIS ABSTRACT HANOI – 2019 THIS STUDY WAS COMPLETED AT THE MILITARY INSTITUTE OF TRADITIONAL MEDICINE Science instructors: Professor, Medicine Doctor NGUYEN MINH HA Associate Professor, Medicine Doctor DINH VAN HAN Reviewer 1: Assoc Prof, PhD Le Luong Dong Reviewer 2: Assoc Prof, PhD Pham Dang Ninh Reviewer 3: Assoc Prof, PhD Nguyen Tran Thi Giang Huong The thesis is protected by The National Council at The Military Institute of Traditional Medicine At (time) date month 2019 YOU CAN READ THIS THESIS IN: - National Library Military Institute of Traditional Medicine Library INTRODUCTION Reason for choosing this subject In daily life, wound always account for a significant proportion due to traffic accidents, labor accidents and domestic accidents The good management of wounds will be the basis for treating other wounds such as bone, nerve and blood vessel wounds In addition, along with the development of the socio-economy, the average life expectancy of people has improved markedly, thereby increasing the chronic diseases, leading to more and more chronic wounds In addition, the situation of antibiotic resistance is becoming more and more serious and concerned by many scientists, the study of a new antibacterial drug is very necessary, in which antibacterial drugs of herbal origin are be concerned LT extract is produced from melaleuca trees for the purpose of treating acute and chronic wounds, with a simple production process, a rich supply of medicinal herbs can be an ideal solution to meet the above requirements Objectives of thesis 1- Research on acute toxicity and subchronic toxicity of LT extract on experimental animals 2- Evaluate the effectiveness of LT extract in the treatment of wounds on experimental and clinical Scientific and practical significances Demonstrate the safety of LT extract Given the hypothesis of the mechanism of action of LT extract according to Traditional Medicine and Modern Medicine in the treatment of wounds Provide a simple, easy to produce form of medicine, can be widely applied in isolated conditions due to war or natural disasters The topic has successfully applied experimental wound research model on experimental animals as a basis for similar studies 4 The structure of the thesis Our thesis consists of 133 pages: Introduction (2 pages); Background (33 pages); Method (24 pages); Results (39 pages); Discussion (32 pages); Conclusion (2 page); Request (1 page) There are 158 references (Vietnamese: 40, English: 99, Chinese: 18, spanish: 1); 51 tables, diagram, chart, 59 figures, 15 appendixes in the thesis Chapter 1: BACKGROUND 1.1 Modern medical concept about wounds 1.1.1 Wound Wounds are defined as the disruption of the continuity of bodycovered tissues, often related to the loss of organization Wounds are classified into main categories: Mechanical wounds, thermal and chemical wounds, ulcers 1.1.2 Physiological of wound healing The normal course of the wound healing process takes place in a sequence of successive separate stages, but there are interwoven phases between stages: hemostasis / inflammation, proliferation and remodeling Diagram 1.1: The process of wound healing 1.1.3 Methods for treating wounds In the treatment of wounds, we must pay attention to two problems, systemic treatment and topical treatment In which the systemic treatment is to ensure the conditions that promote the healing process, eliminate the risk factors of slow wound healing Topical treatment goal is to clean the wound, remove dead tissue and necrosis, excessive secretions, which can cause infection as well as hinder the growth of granulation tissue and epithelium The main methods of wound treatment include: Surgical, Hyperbaric Oxygen Therapy, VAC, growth factors, laser 1.2 Traditional medicine treating wounds Traditional medicine considers that factors affecting the healing process mainly include “qi zhi xue yu”(blood stasis), “ying qi bu cong”(stagnation of ying-qi), “jing luo zu zhi”( blockage of channel), function of viscera are not completed, these factors cause “du re”( toxic heat), necrotic tissues, stasis, deficiency, they interact with each other interfering with the healing process Thereby the treatment can be reduced to main methods: clearing heat, resolve stasis, removing necrotic tissues, growth of new skin These methods mainly use external drugs, so clinically often add strengthen spleen of drugs to increase the effectiveness of treatment 1.3 Some experimental wound models There are many models proposed to study wounds healing on experiment, such as the model of causing the whole skin loss, the model causing burn injury, the model causing chronic wound, model on regenerated skin from tissue culture 1.4 Application of melaleuca in treating wounds Melaleuca cajuputi as well as other melaleuca species, contain many ingredients that have been shown to be effective in treating wounds The process of extracting Melaleuca leaves will yield main products: * Essential oil: The main ingredient is 1,8-Cineol and 4Terpinenol, has antibacterial and anti-inflammatory effects * Melaleuca extract: There are Flavonoid, Ellagic acid, Anthraquinone, all have antibacterial and anti-inflammatory effect LT extract consists of two main parts, essential oils and extracts, from which we assess that LT extract also has antibacterial and antiinflammatory effects Chapter 2: MATERIALS, SUBJECTS AND METHODS 2.1 Material and research equipment - LT extract: LT extracted from Melaleuca cajuputi leaves collected in Phu Quoc, bottled 240ml, the ratio of 1: (1g medicinal / 1ml LT extract) Standard facility - 1% Sulphadiazine silver cream (Silver sulfadiazine) produced by Hue Central Pharmaceutical Company for use as a control drug 2.2 Research subjects Study on acute, subchronic dermal toxicity and experimental wound models in rabbits: 138 white rabbits of Newzealand strain both healthy breeds, weighing 1.8-2.2kg, by the Center Breeding Testing Institute provides Study on antimicrobial susceptibility testing: Using strains of bacteria Pseudomonas aeruginosa, Staphylococus aureus and Escherichia coli Clinical study: 83 inpatients with acute and chronic wounds treated at Military Institute of Traditional Medicine and the National Burns Hospital 2.3 Research Methods 2.3.1 Study acute and subchronic dermal toxicity of LT extract Study acute dermal toxicity according to OECD guidelines and Ministry of Health Acute experimental wounds toxicity under the guidance of the Chinese Ministry of Health Study subchronic dermal toxicity according to OECD guidelines and Lori O Lim 2.3.2 Evaluate the effectiveness of LT extract on experimental and clinical 2.3.2.1 The effectiveness of LT extract in experimental wound treatment * Antimicrobial susceptibility testing According to the method of Vanden Bergher and Vliet link * Effect of treating experimental wounds: Causing experimental wounds: Refer to the guidance of methods to model experimental wounds of Duong Thuy (2003) and of the Department of Labo to study and apply the treatment of burn wounds - National Burn Hospital treatment for 14 days Each rabbit has experimental wounds, one wound is treated with LT extract (zone A1 and A2: research wound), the other wound will be used as a control group (control wounds) Control wounds (30 rabbits) divide the group as follows: Group (Zone B1, n = 10): treated with NaCl 0.9% Group (Zone B2, n = 20): Apply 1% Sulphadiazine silver Evaluate the effectiveness of treatment: Monitor general condition, weight, serum biochemistry test and hematology tests Evaluate the histopathological and microbiology of wound 2.3.2.2 Evaluate the effect of wound treatment of LT extract in clinical clinical trial, longitudinal and cross-sectional study The data were analyzed at pre-treatment times, after days and after 14 days * Treatment according to grouping 17 inpatients with acute wounds randomly divided into groups, research group patients, control group patients Similarly, 66 inpatients with chronic wounds were also randomized to groups, research group 33 inpatients, control group 33 inpatients - Research group: treatment with LT extract ratio 1: - Control group: treatment with silver sulphadiazine 1% Wounds are dressing once a day for 14 days * Evaluate treatment results - Monitor systemic conditions, serum biochemistry test and hematology tests - Compare treatment results between groups based on the criteria of changing wound area, wound healing speed, RESVECH 2.0 scale Microbiological testing, histopathological and ultrastructure testing 2.3.3 Data processing Research results are synthesized and processed by Microsoft Excel 2010 and SPSS 20.0 Compare parameter variables by t-test, non-parameter variables by Fisher test, χ2 test 2.3.4 Medical ethics in research Providing sufficient information related to the study, patients are free of drugs and tests To be considered by the Scientific Council of the Military Institute of Traditional Medicine and the National Burns Hospital Le Huu Trac in terms of ethics and acceptance Chapter 3: RESULTS 3.1 acute and subchronic dermal toxicity in experimental rabbits - Acute dermal toxicity and skin irritation: Both in dermatological and experimental wound studies, rabbits developed normally Because there are no dead rabbits, LD 50 of LT extract has not been determined with the highest dose of 1500mg / kg of weight on the skin pathway - Subchronic dermal toxicity: After weeks and weeks, the rabbit developed normally, there was no special expression The function of hematopoietic function and liver and kidney function were not abnormal The resulting anapathology of the liver, kidneys and skin does not detect damage 3.2 The effect of LT extract in wound treatment on experimental and clinical 3.2.1 The effect of LT extract in wound treatment on experimental 3.2.1.1 Antimicrobial susceptibility testing At 1/4 ratio, LT extract has bactericidal effect on S aureus and P aeruginosa, has bacteriostatic effect on E coli Table 3.10 Antibacterial effect of LT extract S aureus P aeruginosa LT1 1/8 1/4 MBC LT2 1/16 1/4 LT3 1/8 1/4 LT1 1/16 1/4 MIC50 LT2 1/32 1/4 LT3 1/16 1/8 E coli 1/2 1/2 1/2 1/4 1/4 1/4 Bacterial strain 3.2.1.2 Effect of experimental wound treatment * Experimental wound development Table 3.15 Speed of wound healing in rabbits Speed of wound healing (cm2/day) Day zone A1 (LT extract) (n=10) D0-D7 2,31±0,14 2,17±0,13 2,25±0,34 2,31±0,32 D7-D14 0,57±0,045 0,52±0,052 0,55±0,11 0,57±0,10 D0-D14 1,44±0,078 1,34±0,075 1,4±0,17 1,44±0,16 p zone B1 zone A2 zone B2 (NaCl 0,9%) (LT extract) (Sulphadiazin (n=10) (n=20) silver) (n=20) pA1-B10,05 pA1-A2>0,05 pA1-B10,05 pA1-A2>0,05 pA1-B10,05 pA1-A2>0,05 The Speed of wound healing in zone A1 is faster than B1 (p 0.05) * Experimental wound bacteria Table 3.17 Rate of Microbiological testing positive of wounds zone A1 (LT extract) (n=10) zone B1 (NaCl 0,9%) (n=10) zone A2 (LT extract) (n=20) zone B2 (Sulphadiazin silver) (n=20) amount % amoun t % amount % amount 70 60, 17 85 18 D7 20 70, 30 D14 10 60, 10 p0-14 0,05 0,05 90,0 pA2-B2>0,05 pA1-A2>0,05 pA1-B10,05 pA1-A2>0,05 pA1-B10,05 pA1-A2>0,05 0,05 pA1-A2>0,05 D7 402,5±233,04 541,25±181,77 435,0±272,79 D14 90,0±54,08 381,79±128,36 92,5±53,46 Day p zone B2 (Sulphadiazin silver) (n=20) p pA1-B10,05 102,5±60,48 pA1-A2>0,05 pA1-B10,05 pA1-A2>0,05 p0-7
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