The Synthesis, Antibacterial Activities and the Inhibition of Human Malignant Melanoma Cells of Hyaluronic Acid Silver Nanoparticles

• Main Authors: Chieh-Yao Lu, 呂傑堯
• Other Authors: Yang-Chia Shih
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/46132269099924976339

碩士 === 亞洲大學 === 生物科技學系碩士班 === 99 === In recent years, the synthesis of metal nanoparticles is a popular research field. The metals are often used including: gold-nano, silver-nano, iron-nano particles. In particular silver ions, due to they have good electrical conductivity, chemical stability, catalytic and antibacterial activity, so they are often made of silver nanoparticles. They can be used in anti-bacterial, deodorizing, anti-inflammatory and promote wound healing and so on. There are many different methods for the preparation of nanoparticles, in which chemical reduction method is the most commonly used method. The most important three conditions for the preparation of nanoparticles by chemical reduction method include: protective agent, reducing agent and metal salt concentrations. Protective agent used in this study was hyaluronic acid (HA); reducing agent was sodium borohydride (NaBH4) and metal salts of silver nitrate (AgNO3). Hyaluronic acid is common biopolymers in vertebrate. Because the structure of HA is rich in OH group and it carries a large number of negative charged. The moisture effect of HA is very strong and it can reduce the activity of oxide (ROS) of cell damage. The characteristics of HA depends on the different molecular weight, so HA is widely used in biomedical, drug delivery and cosmetic products. The main purpose of this study is to synthesize HA-Ag nanoparticles and to study their biological effects and potential applications. This study is divided into three parts: (1) To synthesize HA-Ag nanoparticles and to investigate the concentration of reducing agent, hyaluronic acid and silver nitrate concentration to find out the best synthesis conditions. The physical properties of HA-Ag nanoparticles included: stability, particle size and optical characteristics were also been analyzed. (2) To evaluate the antimicrobial activity of HA-Ag nanoparticles to E. coli and Staphylococcus epidermidi. (3) To evaluate the inhibition effects of HA-Ag nanoparticles to human malignant melanoma cells. The first part of the study showed that reducing agent (NaBH4) was able to control the size of HA-Ag nanoparticles. Its optimum concentration of NaBH4 was 0.002M, which produced the 15nm size, 398nm of absorption of HA-Ag nanoparticles. If the concentration of reducing agent increased to 0.01M, and the HA-Ag nanoparticles sized up to 21nm and the absorbance value shifted to 404nm. In the second part of experiments, 0.26mg/ml of AgNO3, 0.05 mg/ml of HA and 0.002M of NaBH4 were the best condition for the synthesis of HA-Ag nanoparticles to inhibit E. coli and S. epidermidi, and the largest inhibition diameter was 14mm. The results of the inhibition effects of HA-Ag nanoparticles by the second method indicated that both pathogens were totally inhibited by the nanoparticles which concentration was diluted to 45 μg/ml of AgNO3. The third part of this experiment was to evaluate the inhibition effect of HA-Ag nanoparticles to human malignant melanoma cells. The concentration of AgNO3 and NaBH4 were fixed and prepared with two different HA concentrations to produce HA-Ag nanoparticles as stock. The stock solution was then diluted into 5 different concentrations to treat cells for 24, 48 and 72 hours. The results indicated that the dilution of 8 μg/ml of AgNO3, the inhibition rate of human malignant melanoma cells reached 90%. The inhibition rate increased along with the raise of cultivation time, and the melanoma cells almost died after 72 hours. In summary, according to the above results, they were provide the strong evidences for the inhibition effects of HA-Ag nanoparticles to pathogens and human malignant melanoma cells. In the future, HA-Ag nanoparticles can be used to treat different pathogens or tumor cells for further investigation. Due to HA-Ag nanoparticles have better inhibition ability then Ag+, they can be applied in the development of anit-bacterial biomaterial products. In the future, HA-Ag nanoparticles must have the high potential commercialization and the strong industrial competitiveness.