Nano Carriers for Drug,gene,and Protein Delivery

The  present  research  was  aimed  to  develop  the  meclizine
hydrochloride-polyethylene  glycol  20000  solid  dispersions  to
enhance the solubility and dissolution rate. They were prepared using
solvent  evaporation  method  and  evaluated  for  solubility  studies,
drug-carrier  compatibility  studies  and  in  vitro  dissolution  studies.
From  the  solubility  studies,  formulation  F4  was  selected  to  prepare
the  fast  dissolving  tablets  and  compared  with  control  tablets
(conventional  tablets  using  pure  drug).  From  the  results  of  in  vitro
dissolution  study,  tablets  containing  polyethylene  glycol  20000
showed almost complete drug release within the 15 min. The percent
drug  release  in  15  min  (Q15
)  and  initial  dissolution  rate  for
formulation  F4  was  98.46±1.24%,  6.56%/min.  These  were  very
much higher compared to control tablets (32.49±1.29 %, 2.17%/min).
The  relative  dissolution  rate  was  found  to  be  3.03  and  dissolution
efficiency was found to be 54.44 and it is increased by 3.4 fold with
F4  formulation  compared  to  control  tablets  (16.55).  Thus  the
formulation  of  polyethylene  glycol  20000  solid  dispersions  is  a
suitable  method  to  enhance  the  solubility  and  dissolution  rate  of
meclizine hydrochloride.

Several scientific hurdles still have to be overcome before gene therapy becomes a reality. One of them is the development of safe and efficient gene delivery system. Here, we have employed factorial design to optimize the production of solid lipid nanoparticles (SLN) for gene delivery. A 2×3 full-factorial experimental design was used for the optimization of SLNs formulations. The variables were defined by the components of the formulation: concentration of stearic acid, DOTAP, and Pluronic F68 at two levels (−1, 1) and 3 central points (0). Different SNL formulations were prepared by varying the amount of components and several properties were tested, including their capacity to accommodate DNA and protection against DNase degradation, colloidal stability, in vitro cytotoxicity, and transfection efficiency in prostate cancer cells. Finally, response Surface Methodology was used to select the most effective formulation for gene delivery to prostate cancer cells in vitro. In conclusion, this study revealed that stearic acid and Pluronic F68 were determinant to SLN size and stability, respectively, while small amounts of DOTAP are essential for a successful transfection.

The present work was aimed to design & characterize an oral dosage form to release Terbutaline sulphate following a programmed time period (pulsed release system). Pulsatile release tablet comprises a drug containing core and pH sensitive polymeric coating capable of delaying drug release and providing gastric resistance .The core tablets of terbutaline sulphate were prepared for the treatment of nocturnal asthma .The core tablets were prepared by direct compression method using different disintegrating agents. The cores were coated with pH sensitive polymers (Eudragit S-100, Eudragit L-100) at different coating levels to develop a suitable dosage form which should show minimum drug release in upper regions of gastrointestinal tract (GIT). Prepared tablets were characterized for various physical parameters such as hardness, thickness, weight variation disintegration test drug content and in-vitro drug release characteristics. All the parameters were found to be in the standard range. The dissolution of best formulation F 4 S 3 , F 7 S 3 and F 11 S 3 have shown the lag phase of 5 hrs at 10% coating level and almost complete drug release was achieved after 11 hrs . The kinetic study data for best formulation followed zero order kinetics. Stability study of the best formulation indicates no significant difference in release profile after a period of 3 months.

The objective of the research was firstly to investigate the behavior of Surelease (release retardant binder) alone and in combination with other hydrophilic polymers on the release of zidovudine from sustained release matrix tablet. Secondly, to compare between the release pattern of polyethylene oxide and Methocel K100M alone and in combination. A randomized surface methodology (RSM) was applied to study the effect of polymers on drug release. The independent variables of the formulation were: the type of polymers (X1), concentration of polymers (X2) in the tablet and dependent variables are the percentage drug release at 2 hours (Y1) and the percentage drug release at 12 hours (Y 2 ). Release kinetics were analyzed using zero-order, first order, higuchi's square root and korsmeyer-peppas empirical equations in terms of r 2 . Results of invitro drug release kinetics study suggests that, all formulations follows zero order kinetics with r 2 value in range of 0.99 and fitting the release data to korsmeyers equation release exponent n ranged from 0.83 to 1.22 and followed non-Fickian diffusion mechanism (anomalous transport) and super case II transport for Surelease formulations. Among all the formulations, F14 matrix tablet containing combination of Surelease and polyoxWSR301 shows 102.37% of drug release at the end of 12 hours.

Finding new cancer treatments based on novel drug delivery approaches is critically important.Gold nanoparticles (AuNPs) for use in drug delivery and cellular imaging were functionalized using a two-step synthesis method using modified carboxylic terminated alkane-thiols to attach fluorescent dyes (DAPI or pyNH 2) to the carboxylic ends of the thiolated AuNPs.The cellular cytotoxicity showed that the fluorescently labelled AuNPs were non-toxic at low concentrations.Confocal laser scanning microscopy (CLSM) showed an efficient internalization and uptake of AuNPs by T47D breast cancer cells within 1 hour of incubation.The fluorescence intensity increases over time for both conjugates.When conjugating DAPI with gold nanoparticles, the behaviour is quite different from free DAPI.TEM results showed the localization of AuNPs in the cytoplasm, the cell periphery and near the perinuclear region.The surface chemistry of AuNPs plays an important role in facilitating their cellular uptake as TEM images for conjugates containing DAPI show more AuNPs were internalized by the cells.These various forms of imaging give a clear insight into how these potential drug carriers interact with cells and are subsequently adsorbed into the cells.These are the core processes that will determine if the drug delivery approaches will ultimately be successful.

The drug binding to serum protein (SP) is an area of intense research in evaluating drug candidates. Thus, composite material based on chitosan (CS), and amoxicillin tri hydrate (AMOT) were designed in order to evaluate the release of AMOT@CS NPs and it's binding with BSA under physiological condition. The inclusion of 2% v/v T80 in the CS NPs enabled in vitro release of AMOT, at 298 K and pH 7.4 up to 68.03%. This study also was undertaken to compare and explore the binding of potential chemotherapeutic antibacterial drug Amoxi-cillin trihydrate (AMOT) and AMOT@Chitosan nanoparticles (AMOT@CS NPs) with a model protein bovine serum albumin (BSA) by fluorescence spectroscopy, synchronous fluorescence spectroscopy, Ultraviolet-visible (UV-vis) absorption, and CD spectroscopic techniques. Experimental results indicated that AMOT and AMOT@CS NPs could bind with BSA and quench the fluorescence of BSA via static mechanism. The interactions among BSA with AMOT, AMOT/T80 and AMOT@CS NPs were evidenced by substantial changes in the BSA secondary structure, as revealed by circular dichroism. The findings of thermodynamic parameters revealed that the binding reaction in both systems was exothermic and spontaneous, enthalpically driven and that the hydrogen bonding and Vander Waals forces played a vital role to achieve optimal interaction between AMOT/ AMOT@CS NPs and BSA.

Objective: The present investigation aimed at phytochemical screening of the whole plant of Anoectochilus formosanus Hayata (Orchidaceae) after successive extraction followed by hepatoprotective activity of its aqueous extract. The research work also focused on the sub acute toxicity study of aqueous extract of the same plant.
Methods: Successive extraction was carried out with petroleum ether, chloroform, methanol and water respectively. Hepatoprotective activity of its aqueous extract was investigated in carbon tetrachloride, ethanol and paracetamol induced hepatotoxic rat models and compared with silymarin (20 mg/kg) as reference standard. Sub acute toxicity study was elicited by studying the effect of Anoectochilus formosanus on the lipid profile, biochemical parameters and hematological parameters in rats and compared with standard silymarin (20 mg/kg).
Results: Phytochemical screening revealed the presence of anthraquinone glycosides, cardiac glycosides, reducing sugars, carbohydrates, phenolic compounds, tannins, flavonoids and saponins in the aqueous extract of the plant under investigation. Two way analysis of variance study of the estimated biochemical parameters for instance, aspartate aminotransferase, alanine amino transferase and alkaline phosphatase were revealed that there is significant difference (p-value < 0.0001) exists between the different treatment groups supported by least significant difference test of various biochemical parameters, which was also evident from the histopathological study of liver sections. Sub acute toxicity study revealed no significant toxic effects.
Conclusion: Aqueous extract of Anoectochilus formosanus (200 mg/kg) had shown significant hepatoprotective activity as compared to standard silymarin. Sub acute toxicity studies had concluded that it might be considered safe for a longer duration of time.

The purpose of the presented study here was to evaluate the prevalence of hepatitis C and B infections in patients diagnosed with or suspected of hepatic carcinoma or any other co-morbid with the objective of correlating diagnostic efficacy of alpha-fetoprotein (AFP). The study covers the period of January 1998 to January 2008. Study requirements such as medical history, reports and related information of patients from both genders (n = 93) with 50 males and 43 females were gathered for analysis and assessment ensuring that diagnostic confirmation of HCC existed with lab test of alpha-feto proteins available. Results shows the distribution analysis of hepatitis infections, the related co-morbid, the high-level AFP results and positive hepatitis profile as; hepatic carcinoma with HCV origin 33.33% (n = 31) and with HBV origin 41.93% (n = 39). For HCC of cirrhotic origin only, 10.75% (n = 10) was noted with no-infection whereas HCC of chronic liver disease [CLD] was around 13.97% (n = 13). As per medical setting (e.g. cirrhosis and chronic liver disease) and hepatic infections (B and C), 50 males patients were segregated as carcinoma patients with HCV and HBV, n = 15 and 24, whereas with cirrhosis and CLD, 6 and 5 respectively. Forty three female patients were also segregated in similar manner, thus resulting in groups of N = 14, 17, 5 and 7, respectively for HCV and HBV related carcinoma and non-malignant co-morbid such as cirrhosis and chronic liver disease. Regarding AFP levels, in males, 10.00% (n = 10) patients have AFP in the highest range of > 300.00 to < 768.00 ng/ml in HCV group; 46.00% (n = 23) in the range of > 150 to < 300 ng/ml in HBV and cirrhosis groups and 8% (n = 4) in the range of >300 to <520 ng/ml in HBV alone group. In females, 20.90% (n = 9) showed AFP ranges between >300 to 612 ng/ml in HCV group; and Address for Correspondence 18.60% (n = 8) in range of >150 to <300 ng/ml in HBV and CLD groups. Nonetheless the present studies neither irrevocably instate direct correlation between presence of hepatitis infections and formation of HCC nor ascertain that hepatitis infection were the 'only' cause of HCC. However efficacy of AFP has been found to be significant as a diagnostic tool where HCC was related to the hepatitis infections. The analytical data which was assessed after combining all factors is demonstrated here according to medical conditions diagnosed, risk factors obtained and the etiological factors related to the carcinoma and alpha-feto protein concentration characteristics.

This work evaluated under accelerated conditions the stability of the
biodegradable gentamicin-glyceryl monooleate water based-gel used
in the treatment of chronic osteomyelitis. Three batches of 600.0 g of
gentamicin -glyceryl monooleate -water based-gel (5/80/15% w/w)
were manufactured. The samples were stored in a climatic test
chamber for stability evaluation according to the standards published
by the International Conference on Harmonisation and the World
Health Oganization (25 ± 2°C / 60 ± 5%RH) for 6 months. Samples
were evaluated at pre-determinate times 0, 3 and 6 months with
regard to their physicochemical properties and In Vitro drug release
characteristics. Gentamicin sulfate present in the freshly prepared
gels was the same after three and six months of storage at 25°C / 60%
RH. However the relative amount of gentamicin sulfate in the freshly
prepared gel that was about 110% decreased by about 11% after six
months of storage. The release profiles of the gels remained similar
despite a partial hydrolysis of glyceryl monooleate and an increase in
the residual amount of water and in the viscosity.

Calcium phosphate nanoparticles present a unique class of non-viral vectors, which can serve as efficient and
alternative DNA carriers for targeted delivery of genes. In this study we report the design and synthesis of ultra-low
size, highly monodispersed DNA doped calcium phosphate nanoparticles of size around 80 nm in diameter. The DNA
encapsulated inside the nanoparticle is protected from the external DNase environment and could be used safely to
transfer the encapsulated DNA under in vitro and in vivo conditions. Moreover, the surface of these nanoparticles
could be suitably modified by adsorbing a highly adhesive polymer like polyacrylic acid followed by conjugating the
carboxylic groups of the polymer with a ligand such as p-amino-1-thio-b-galactopyranoside using 1-ethyl-3-(3-
dimethylaminopropyl)-carbodiimide hydrochloride as a coupling agent. We have demonstrated in our studies that these surface modified calcium phosphate nanoparticles can be used in vivo to target genes specifically to the liver.

a b s t r a c t Nanocarriers (NCs) are colloidal systems having structures below a particle or droplet size of 500 nm. In the previous years, the focus for the application of NCs was primarily placed on the parenteral and oral application. However, NCs applied to the skin are in the center of attention and are expected to be increasingly applied as the skin offers a lot of advantages for the administration of such systems. For the use of NCs to the skin, one has to differentiate between the desired effects: the local effect within the skin (dermal drug delivery) or a systemic effect accompanied by the permeation through the skin (transdermal drug delivery).

Polyamidoamine dendrimers (PAMAM) are being used as efficient vectors for delivery of nucleic acids to the cells. However, these dendrimers cause a significant amount of cytotoxicity. In order to improve its transfection efficiency and cell viability, surface amine groups of PAMAM were converted into guanidinium (Gn) and tetramethylguanidi-nium (TMG) moieties. These modified PAMAM dendrimers interacted with negatively charged plasmid DNA efficiently and formed stable complexes as revealed by dynamic light scattering analysis. PAMAM/pDNA, PAMAM-TMG/pDNA and PAMAM-Gn/pDNA complexes were found to be in the range of 175-250 nm with zeta potential in the range of +21-37 mV. Further, these modified dendrimers did not display toxicity rather it decreased a bit when tested in HEK293, HeLa and MCF-7 cells. Among these modified dendrimers, PAMAM-Gn/pDNA complex displayed the highest transfection efficiency in both the cell lines HEK293 and MCF-7.

Different techniques have been employed to produce chitosan nanoparticles; thus, suitable alternatives and easy-handling production methods are highly desired. We used a tank reactor with baffles and mechanical stirring with Cowles impellers that can feasibly be up-scaled and allows for the production of homogenous chitosan nanoparticles. First, we explored the effects of temperature on the produc- tion of chitosan (CHI)/pentasodium tripolyphosphate (TPP) nanoparticles. We compared the effect of gradual temperature decrease between 40 to 55◦C with the effect of isothermal processes (25, 45 and 55◦C). CHI/TPP nanoparticles produced by the gradual temperature decrease technique had polydis- persity indices (PDIs) that were significantly smaller than the nanoparticles produced by isothermal processes. To further control the production process, we employed an experimental design to determine the effects of CHI and TPP concentrations and CHI/TPP mass ratios in the gradual temperature decrease technique. Under all conditions, the sizes of the CHI/TPP nanoparticles were on the nanoscale. Gene deliv- ery capacities of the nanoparticles were evaluated; the plasmid pEGFP-N1 was incorporated (10 and 40% of the chitosan weight) into the nanoparticles and incubated with 293A cells. The in vitro transfection efficiencies were evaluated over 120 h and indicated sustained gene delivery at 10% pDNA. The poly- plexes demonstrated some cytotoxicity after 120 h. These findings will contribute to the development of processes for generating CHI/TPP nanoparticles for gene delivery applications.

Almond gum (Badam Gum/ Badam Pisin) is a natural polymer composed of mainly carbohydrates (>90%) which makes it a potential candidate in targeted drug delivery. It contains 46% L-arabinose which is known to have fluid entrapment capacity. Various methods of drug targeting against tumor have been developed but due to poor solubility in body fluids, the bioavailability of the drug is reduced and so is its effectiveness. Almond gum is known to be water-soluble so the required bioavailability will be achieved and the drug dosage will also be minimal. Since it is of natural origin side effects will be less compared to that of artificially prepared drugs. Studies show that almond gum is known to have various health benefits including increasing immunity and acting as a coolant. Terminalia catappa (Indian almond), the source of almond gum is abundant in the Indian subcontinent hence making it more economical than chemical surfactants. The objective of this study is to determine whether Almond gum has a higher drug entrapment efficiency than that of Niosomes.