IN VITRO STUDY OF THE EFFECT OF PHYLLANTHUS NIRURI EXTRACT-LOADED CHITOSAN NANOPARTICLES ON CLDN3 GENE EXPRESSION IN TM4 CELLS

• Main Author: Mutiara, Adinda
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/78407
• Institution: Institut Teknologi Bandung

The application of nano-systems in the pharmaceutical is using Phyllanthus niruri (P. niruri) extract as a co-adjuvant loaded in a chitosan nanoparticle carrier system for use as an oral immunostimulant. Despite the promising innovations in the pharmaceutical field, there are emerging toxicities that have been studied before. Previously it was known that chitosan nanoparticles containing P. niruri extract had been tested in vivo and in vitro that the formulation was toxic to the process of spermatogenesis which has the potential for infertility in men. Nanotoxicology studies are rarely studied. This study aims to examine the effect of P. niruri extractloaded chitosan nanoparticles (NP PN), blank chitosan nanoparticles (K NP), and P. niruri extract (E PN) on the expression of Cldn3 and Stra8 genes. The study began with NP PN and NP K formulations, characterized, and tested for analysis of Cldn3 and Stra8 gene expression from TM4 cells by qPCR. The results of the characterization of PN NPs, as particle size, polydispersity index, and zeta potential were respectively 155.87 nm; 0.30; +32.54 mV. Meanwhile, the NP K is 163.97 nm; 0.26; +30.54 mV with entrapment effiency is 87,83% meets the optimal criteria to be delivered into the body. Primary quality analysis including preliminary test, primer specificity, and primary efficiency test showed optimal results for Cldn3 as target gene and Gapdh(2) as housekeeping gene for gene expression analysis at 60°C annealing temperature. Stra8 could not be analyzed for gene expression because the primers were non-specific and did not have optimal primer efficiency. Based on the results of analysis of Cldn3 gene expression, it was found that administration of NP K did not significantly inhibit gene expression, but PN NP concentrations of 62.5 to 2000 ppm and E PN concentrations of 31,25; 125; 250 ppm significantly inhibited gene expression (P<0.05). It can be concluded that NP PN and E PN without being encapsulated in chitosan nanoparticles have the risk of interfering with the process of spermatogenesis and potentially infertility in men.