Formulation of nanoparticles from short chain chitosan as gene delivery system and transfection against T47D cell line

Recently numerous prototype DNA-based biopharmaceuticals can be used to  control  disease  progression  by  induction  and  inhibitin  the  overexpression  of genes.  Since  there  are  poor  cellular  uptake  and  rapid  in  vivo  degradation  of DNA-based  therapeutics  therefore  the  use  of  delivery  systems  to  facilitate cellular internalization and preserve their activity is necessary. Cationic polymers commonly used as carriers to delivery gene because of easy to form complexes and  higher  stability  compared  to  that  lipoplexs.  Chitosan,  a  cationic,  are polymer most widely used in gene delivery systems because of the low toxicity, and biocompatible. The aim of this study was to formulate nanoparticles of short chain  chitosan-pEGFP-C1  and  short  chain  chitosan/TPP-pEGFP-C1  by coaservation  complex  method.  Stability  test  of  the  formula  was  performed  by incubating the nanoparticles complex with DNase I and Artificial Intestinal Fluid. Cytotoxicity  and transfection  studies  were  evaluated  against  T47D  cell line.  The diameter  of  Chitosan-pEGFP-C1  and  chitosan/TPP-pEGFP-C1  nanoparticles  were on the range of 56–282.8 nm. The zeta potential wasdetermined to be +14.03 - +16.6  mV.  Stability  studies  showed  that  chitosan-pEGFP-C1  and  chitosan/TPPpEGFP-C1  nanoparticles  were  stable,  undegradable  by  DNase  I  and  artificial intestinal fluid. Cytotoxic Assay of Chitosan-pEGFP-C1 and  chitosan/TPP-pEGFPC1  nanoparticles  (pH  4.0)  showed  that  the  viability  of cell  was  >  90%  for  all formulas.  EGFP-C1  plasmid  gene  delivered  by  chitosan  nanoparticles  can  be expressed  in  T47D  cell  culture.  According  to  these  results  chitosan  and chitosan/TPP  nanoparticles  had  potentially  to  be  used  as  a  non-viral  vector system delivery for gene therapy.Key words:Chitosan, Nanoparticles, Plasmid EGFP-C1, Cell culture T47D

SISTEM PENGHANTARAN OBAT TERTARGET, MACAM, JENIS-JENIS SISTEM PENGHANTARAN, DAN APLIKASINYA

Development of methods to improve delivery of drugs for treating dangerous disease like cancer and viral infections are very important today. Selectivity in therapy is needed, hence capability to deliver drugs at specific site being studied in order to reduce toxicity and unwanted side effect at non target site. Many drugs delivery from passive targeting to active targeting delivery will be improved to reach that purpose. Because this system is very important for cancer therapy, many cytotoxic agent have been approved by clinical examination. Besides that targeting drug delivery is very useful for neurological diseases such as alzhemier, liver diseases, kidney, lung, and colon diseases

Review Artikel: PENGGUNAAN FORMULASI NANOPARTIKEL KITOSAN SEBAGAI SISTEM PENGHANTARAN GEN NON VIRAL UNTUK TERAPI GEN

Gene therapy is a method to repair damaged or defective genes that responsible for the onset of certain diseases. Now, numerous prototypes of DNA can control the progress of the disease through induction or inhibition of genes, but the bad cellular uptake and the rapid in vivo degradation of DNA-based therapy require the use of delivery system that can facilitate the cellular internalization and keep its activity. Cationic polymers are commonly used as gene carrier because it is easy to form complexes and it has higher stability compared with lipoplexes. Cationic polymer chitosan is the most widely used in gene delivery system because of the low toxicity andbiocompatible. Gene delivery system using viruses shows high transfection result but it has many disadvantages, such as oncogenic effects and immunogenicity. However, cationic polymers, such as chitosan have the potential for complexation of DNA that can be used as a non-viral vector for gene therapy. Chitosan provides a strongelectrostatic interaction with negative charge of DNA to form nanoparticles and effectively protect from nuclease degradation. Those properties make chitosan become a good candidate from non viral gene delivery

Examination of Analgetics Effect of Extract Boesenbergia Pandurata(roxb.) Schlechter to Swiss Furrow Male Mice

This research is conducted to evaluate analgetics effect of Boesenbergia pandurata (Roxb.) Schlechter extract using Witkin et al method. This method is conducted to see the elimination effect of pain after the administration of acetate acid by intraperitoneal injection to Swiss furrow male mice. Pain effect by the administration of acetate acid causing stomach wall contraction, till the head and feet pulled rear and abdomen touch the cage room base. This symptom is named by writhing reflex and this symptom can be eliminated with an analgesic. Analysis was done by comparing the amount of writhe that happened after the giving of Boesenbergia pandurata (Roxb.) Schlechter extract with acetosal as positive comparator and aquadest as negative control. Writhing effect calculated during 30 minute after the administration of acetate acid by intraperitoneal injection. Boesenbergia pandurata (Roxb.) Schlechter extract between dose 15; 30 and 60 mg / kgBW have significant difference ( P<0.05). In the dose-effect relation it can be seen that more higher the dose give more higher effect. The amount of writhing in Boesenbergia pandurata (Roxb.) Schlechter extract dose 30 mg / kgBW not significantly differ with Acetosal, while Boesenbergia pandurata (Roxb.) Schlechter extract dose 15 mg / kgBW and 60 mg / kgBW have significant difference with Acetosal. Boesenbergia pandurata (Roxb.) Schlechter extract dose 15 mg / kgBB showing the amount of writhing larger than Acetosal and Boesenbergia pandurata (Roxb.) Schlechter extract dose 60 mg / kgBB has amount of writhing fewer than Acetosal. From the calculation of protection percentage administration significant difference between group. Boesenbergia pandurata (Roxb.) Schlechter extract dose 30 mg / kgBW and 60 mg / kgBW do not show significant difference with Acetosal. While from effectiveness percentage analysis also showing significant difference between group except Boesenbergia pandurata (Roxb.) Schlechter extract dose 30 mg / kgBW with Acetosal 130 mg / kgBW. So from overall of analysis resulting that analgetics activity of Boesenbergia pandurata (Roxb.) Schlechter extract start from dose 30 mg / kgBW have effective effect to reduce the pain induced with acetatic acid which equivalent with Acetosal as positive control. More great the dose give higher activity. From this research the dose of Boesenbergia pandurata (Roxb.) Schlechter extract 60 mg / kgBW have analgetics activity stronger than Acetosa

FORMULA OPTIMIZATION OF ORALLY DISINTEGRATING TABLET CONTAINING MELOXICAM NANOPARTICLES

Meloxicam is one of oxicams anti-inflamatory drugs that are effective to relieve toothaches, arthritis, dysmenorrhea, and fever. Meloxicam in this study was milled with High Energy Milling (HEM) method to obtain its nano size and then direct compression method was used to produce Orally Disintegrating Tablet (ODT). ODT is designed to be rapidly dissolved on the tongue within a minute. It can be administered without water or chewing and may improve the bioavailability and effectiveness of the drug, and increase the patient’s adherence. The present study aimed to understand the effects of Ac-Di-Sol and Kollidon CL as superdisintegrants, that were used separately or in combination, on the characteristics of nanoparticles meloxicam ODT. It was also to obtain the best proportion of combination between Ac-Di-Sol and Kollidon CL that can produce the optimum formula of meloxicam ODT. The effects of single or combined superdisintegrants were evaluated using Simplex Lattice Design (SLD). Ac-Di-Sol (X1) and Kollidon CL (X2) were the independent variables, while the dependent variables were friability (Y1), disintegrating time (Y2), wetting time (Y3), and percent meloxicam release after 60 seconds (Y4). Optimization of five nanoparticle meloxicam ODT formulas was conducted using Design Expert 7.1.5. The combination of Ac-Di-Sol 4.05mg (X1) and Kollidon CL 10.95mg (X2) in 150mg nanoparticles meloxicam ODT can produce optimal ODT characteristics. After verification there was no difference between predicted value and observed value with p value &gt; 0.05

MELOXICAM SELF-NANOEMULSIFYING DRUG DELIVERY SYSTEM: FORMULATION AND RELEASE KINETICS ANALYSIS

Objective: This study aimed to find the best SNEDDS meloxicam formula and analyze the release kinetics of meloxicam SNEDDS and non-SNEDDS using DDSolver.&nbsp; Methods: Meloxicam SNEDDS was prepared using sunflower seed oil, Cremophor RH 40 as a surfactant, and polyethylene glycol (PEG) 400 as a co-surfactant.&nbsp; Results: The best formula obtained subjected to the in vitro dissolution study was analyzed using DDSolver. The study shows one selected formula consists of 10% sunflower seed oil, 70% cremophor RH 40, and 20% PEG 400 with a 20.5 nm±12 nm droplet size. The dissolution study showed that SNEDDS could significantly increase the meloxicam release compared to the non-SNEDDS formulation. The kinetics of meloxicam release from SNEDDS formulations follow the Weibull release model (β = 1.00).&nbsp; Conclusion: This study concludes that SNEDDS best prepared in sunflower seeds oil: Chremophor RH 40: PEG 400 ratio of 1: 7: 2 and has the potency to increase the solubility and dissolution of meloxicam

FORMULATION OF SELF-NANOEMULSIFYING DRUG DELIVERY SYSTEM OF BOVINE SERUM ALBUMIN USING HLB (HYDROPHILIC-LYPOPHILIC BALANCE) APPROACH

Self-Nanoemulsifying Drug Delivery System (SNEDDS) has potential to be developed for oral protein delivery because it is free from water, hence preserving the stability of protein, protecting protein from enzymatic degradation, and enhancing the protein permeability in the gastrointestinal tract (GIT). However, protein-based SNEDDS formulation is challenging due to low solubility property of protein in oil, which is towards zero. This present study aimed to obtain the most compatible SNEDDS system for protein using HLB approach. Bovine serum albumin (BSA) was used as a protein model in this study. A number of 78 formulae with HLB ranging between 11 and 15 were screened to acquire stable SNEDDS composition without the presence of phase separation. Of 13 stable formulae, two were selected (F30, F45) with HLB 15, and then loaded with BSA. Physical characteristics of both formulae were then evaluated and the results suggested that SNEDDS with single hydrophilic surfactant (F45) and HLB 15 was the best formula for protein template as the stability testing showed that phase separation and precipitation did not appear. It was robust to pH and dilution with percentage of transmittance of 96.40±1.05% and the droplet size of 180.9nm. F45 also had uniform distribution of droplets size since the polydispersity index was less than 0.1. The zeta potential of F45 was -0.12mv with loading efficiency 83.57±1.77%. The emulsifying time of F45 was &gt; 2min due to the formation of crystalline gel that was difficult to disperse

Formulation of Insulin Self Nanoemulsifying Drug Delivery System and Its In Vitro-In Vivo Study

Particulate delivery system can be used for improving the efficacy of protein and peptide drug. In addition to a polymer-based particulate delivery system, self-nanoemulsifying drug delivery system (SNEDDS), a lipid-based delivery system, is currently developed for either less water-soluble or soluble drugs. This study aims to design SNEDDS for oral insulin administration and its in vitro-in vivo study. The SNEDDS template was designed using D-optimal mixture design and was analyzed using software Design Expert 7.1.5. The obtained optimum template was loaded with insulin and evaluated for its transmittance percentage, emulsification time, particle size, zeta potential, stability, the amount of insulin in vitro diffused across rat intestine, and insulin serum concentration after oral administration. The study results revealed that the optimum template of SNEDDS formula consisted of 10% (w/w) Miglyol 812N, 65% (w/w) Tween 80, and 25% (w/w) propylene glycol. These optimum template then was loaded with insulin and characterized. SNEDDS insulin has particle size of 12.0±1.7 nm, zeta potential of +0.16mV, transmittance of &gt;90%, and emulsification time of &lt; 60 seconds. The stability study showed that SNEDDS insulin was stable from both precipitation and phase separation. The amount of insulin transported from SNEDDS formula in vitro was 32.45±2.03% and non-SNEDDS formula was 10.44±5.04%. In vivo study of SNEDDS insulin produced a significantly increased Cmax, AUC, and F value than insulin non SNEDDS (p &lt; 0.05). In brief, SNEDDS formulation in this study is a promising approach to increase the effectiveness of oral insulin. Insulin is better given orally in SNEDDS formulation than in non SNEDDS formulation

UJI EFEK ANALGETIKA EKSTRAK RIMPANG TEMU KUNCI (Boesenbergia pandurata (Roxb.) Schlechter PADA MENCIT JANTAN GALUR SWISS

This research is conducted to evaluate analgetics effect of Boesenbergia pandurata (Roxb.) Schlechter extract using Witkin et al method. This method is conducted to see the elimination effect of pain after the administration of acetate acid by intraperitoneal injection to swiss furrow male mice. Pain effect by the administration of acetate acid causing stomach wall contraction, till the head and feet pulled rear and abdomen touch the cage room base. This symptom is named by writhing reflex and this symptom can be eliminated with an analgesic. Analysis was done by comparing the amount of writhe that happened after the giving of Boesenbergia pandurata (Roxb.) Schlechter extract with acetosal as positive comparator and aquadest as negative control. Writhing effect calculated during 30 minute after the administration of acetate acid by intraperitoneal injection. Boesenbergia pandurata (Roxb.) Schlechter extract between dose 15; 30 and 60 mg / kgBW have significant difference ( P<0.05). In the dose-effect relation it can be seen that more higher the dose give more higher effect. The amount of writhing in Boesenbergia pandurata (Roxb.) Schlechter extract dose 30 mg / kgBW not significantly differ with Acetosal, while Boesenbergia pandurata (Roxb.) Schlechter extract dose 15 mg / kgBW and 60 mg / kgBW have significant difference with Acetosal. Boesenbergia pandurata (Roxb.) Schlechter extract dose 15 mg / kgBB showing the amount of writhing larger than Acetosal and Boesenbergia pandurata (Roxb.) Schlechter extract dose 60 mg / kgBB has amount of writhing fewer than Acetosal. From the calculation of protection percentage administration significant difference between group. Boesenbergia pandurata (Roxb.) Schlechter extract dose 30 mg / kgBW and 60 mg / kgBW do not show significant difference with Acetosal. While from effectiveness percentage analysis also showing significant difference between group except Boesenbergia pandurata (Roxb.) Schlechter extract dose 30 mg / kgBW with Acetosal 130 mg / kgBW. So from overall of analysis resulting that analgetics activity of Boesenbergia pandurata (Roxb.) Schlechter extract start from dose 30 mg / kgBW have effective effect to reduce the pain induced with acetatic acid which equivalent with Acetosal as positive control. More great the dose give higher activity. From this research the dose of Boesenbergia pandurata (Roxb.) Schlechter extract 60 mg / kgBW have analgetics activity stronger than AcetosalPenelitian ini dilakukan untuk mengevaluasi efek analgetik ekstrak Boesenbergia pandurata (Roxb.) Schlechter menggunakan metode Witkin et al. Metode ini dilakukan dengan melihat efek penghilangan rasa sakit setelah pemberian asam asetat secara intraperitoneal kepada tikus jantan galur swiss. Efek rasa sakit yang ditimbulkan akibat pemberian asam asetat akan menyebabkan kontraksi dinding sel perut, sampai kepala, kaki yang tertarik ke belakang dan perut yang menyentuh dasar kandang. Simptom ini dinamakan writhing reflex dan dapat dieliminasi dengan analgetik. Analisis dilakukan dengan membandingkan jumlah writhing reflex (geliat) setelah pemberian ekstrak dengan  asetosal sebagai kontrol positif dan aquadest sebagai kontrol negatif. Efek geliat dihitung 30 setelah pemberian asam asetat dengan injeksi intraperioneal. Efek ekstrak Boesenbergia pandurata (Roxb.) Schlechter antara dosis 15, 30 dan 60 mg/kg BB memberikan perbedaan yang signifikan ( P<0.05). Semakin tinggi dosis, semakin tinggi efek yang diberikan. Jumlah geliat pada dosis 30 mg/kg BB tidak berbeda dengan pemberian  asetosal sedangkan dosis 15 dan 60 kg/kgBB berbeda dengan  asetosal. Pemberian dosis 15 mg/kg BB memberikan jumlah geliat yang lebih besar sedangkan pemberian 60 mg/kg BB memberiakn jumlah geliat yang lebih kecil dari  asetosal. Dari perhitungan presentasi proteksi, terdapat perbedaan antar kelompok tetapi dosis 30 dan 60 mg/kgBB tidak berbeda dengan  asetosal. Analisis presentasi efektifitas menujukkan perbedaan antar kelompok kecuali dosis 30 mg/kgBB dengan  asetosal. Secara keseluruhan dapat dikatakan bahwa efek analgesik ekstrak Boesenbergia pandurata (Roxb.) Schlechter bermula pada dosis 30 mg/kgBB yang setara dengan  asetosal. Semakin tinggi dosis, semakin tinggi aktivitasnya dan pemberian ekstrak 60 mg/kgBB memberiak efek lebih besar dari  asetosal

Optimasi Hydroxypropyl Methylcellulose dan Chitosan pada Tablet Floating-Mucoadhesive Diltiazem Hidroklorida Menggunakan Desain Faktorial

Diltiazem HCl is one of the drugs used for hypertension treatment. It requires frequent dosing, which is why diltiazem HCl needs to be formulated into preparations using a controlled release drug delivery system. The combination of floating and the mucoadhesive system is expected to increase the stomach's retention of the dosage form. This study aimed to determine the optimum composition of hydroxypropyl methylcellulose (HPMC K100M) and chitosan for floating mucoadhesive diltiazem HCl tablet. Tablets that have been prepared were evaluated for the tablet's physical characterization, powder flowability test, dissolution test, floating ability, and mucoadhesive test. Tablets were optimized using a factorial design, and the data were analyzed using design expert 11.0.0. The results showed that the optimum formula for polymer combination in diltiazem HCl tablet was 175 mg for HPMC K100M and 50 mg for chitosan. The combination of polymers with this amount can produce a floating lag time of 45,333 seconds, floating duration time &gt;12 hours, and the strength of mucoadhesive is 81,633 grams