PREPARATION CHITOSAN NANOPARTICLE CONTAIN PHYLLANTUS NIRURI EXTRACT AND IN VITRO TOXICITY ASSAY IN TM4 CEL

• Main Author: Pradana, Reinaldo
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/56675
• Institution: Institut Teknologi Bandung

Application nanoparticles provide promising efficacy in pharmaceutical field. Most of nanoparticles investigation only focus on efficacy nanoparticles on human health. While limited information was available on the toxicity and its toxicity mechanism. Phyllantus niruri is medicinal plant traditionally known as immunomodulatory agent. In previous research, Phyllantus niruri extract was formulated as adjuvant into nanoparticle chitosan contain antigen HBsAg for increased immune response against hepatitis vaccine. It is known that Phyllantus niruri extract was reported has toxic effect on testis specially in sertoli cells and germ cells. There had been no studies related to the toxicity of chitosan nanoparticle containing Phyllantus niruri extract in testis. The research of nanoparticle toxicity research of nanoparticle in testis was limited to inorganic nanoparticle such as ZnO nanoparticle and Ag nanopartikel. Both of them were known to be toxic to testis and disrupt spermatogenesis. This research was undertaken to determine the toxicity of nanoparticle chitosan contain Phyllantus niruri extract (PN NP) and the toxicity mechanism that observed in TM4 cells (sertoli cells). The research concept are formulation and characterization of NP PN and cell viability study using Cell Counting Kit-8 (CCK-8); apoptotic necrotic study using annexin V propidiuim iodide; cell cycle analysis using propidiuim iodide; ROS study using DCFDA. All of characterization of NPs PN met specification, there were particle size around 170 nm, polydispersity index 0,1-0,3, zeta potential +37 mV and entrapment efficiency had good results (79%). Cell viability study used CCK-8 showed that PN inhibit cell growth of Sertoli PN NP contain 7,8 – 2.000 µg/mL Phyllantus niruri extract. Preparation of NP PN also induced apoptotic significantly against control, and cell cycle analysis confirm there was cell cycle arrest at phase of G2/M. ROS analysis also confirm that PN NP caused increasing of ROS in sertoli cells. In conclusion, PN NP was toxic in sertoli cells with the highest cell growth inhibition at dose 125 µg/mL and the toxicity mechanism was induction of apoptotic and also cell cycle arrest at G2/M phase due to increasing ROS.