Papers by Author: Praneet Opanasopit

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Authors: Natthan Charernsriwilaiwat, Theerasak Rojanarata, Tanasait Ngawhirunpat, Praneet Opanasopit
Abstract: The aim of this study was to investigate the antioxidant activities of chitosan acetate (CS-acetate), chitosan hydroxybenzotriazole (CS-HOBt), chitosan thiamine pyrophosphate (CS-TPP) and chitosan ethylenediaminetetraacetic acid (CS-EDTA) nanofiber mats. Chitosan was dissolved with hydroxybenzotriazole (HOBt), thiamine pyrophosphate (TPP) and ethylenediaminetetraacetic acid (EDTA) in distilled water. These chitosan aqueous salts were blended with polyvinyl alcohol (PVA) at volume ratio 30/70 chitosan salts/PVA and prepared to nanofibers via electrospinning process. The morphology of electrospun chitosan aqueous salts based nanofiber mats were observed under scanning electron microscope (SEM). The antioxidant activities were determined employing various established in vitro system such as superoxide, hydroxyl radicals scavenging and metal ion chelating compared with pure PVA nanofiber mats. The results exhibited that the electrospun chitosan aqueous salts based nanofiber mats showed the different antioxidant activity depended on salt forms. Among the salt forms, CS-acetate nanofiber mats showed the highest superoxide radical scavenging effect while CS-HOBt nanofiber mats (IC50 = 7.53 mg/mL) showed the highest hydroxyl radical scavenging effect. For the metal ion chelating activity, CS-EDTA nanofiber mats showed the highest chelating activity (IC50 = 1.07 mg/mL). In summary, the antioxidant chitosan aqueous salt based nanofiber mats have potential for use in pharmaceutical applications.
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Authors: Nanthida Wonglertnirant, Tanasait Ngawhirunpat, Praneet Opanasopit, Hiroaki Todo, Kenji Sugibayashi
Abstract: Delivery of drugs through skin is obstructed by the excellent barrier properties of the outermost skin layer, the stratum corneum (SC). A strategy employing microneedles have recently emerged as a minimally invasive device for disrupting the SC structure and creating holes for molecules to pass through. Hollow-typed microneedles permit drug delivery which can be modulated over time via active delivery controlled by hand or pump. In this study, the potential of hollow microneedle for overcoming the outermost skin barrier and facilitating drug delivery into skin was investigated. Fluorescein isothiocyanate (FITC)-dextrans (4.3 kDa), FD-4, was used as a model large molecular compound. The effects of injection volume and formulation on drug release behavior from skin were determined. FD-4 was favorably loaded into the lower epidermis as well as the superficial dermis of the skin by a hollow microneedle. The release profiles of FD-4 were analyzed by Higuchi model based on Fick’s law of diffusion. The higher the volume of FD-4 solution injected, the faster the FD-4 release rate from skin. Liposome formulation exhibited no difference on drug release profiles compared with the solution. The results provide information for designing an effective hollow microneedles system.
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Authors: Maleenart Petchsangsai, Nanthida Wonglertnirant, Theerasak Rojanarata, Praneet Opanasopit, Tanasait Ngawhirunpat
Abstract: The objective of the present study was to use hollow microneedles for enhancing the transdermal delivery of Bovine Serum Albumin (MW~66,000 Da)-Fluorescein Isothiocyanate (BSA-FITC) conjugate, a hydrophilic large molecular compound. BSA-FITC was successfully loaded into the lower epidermis as well as the superficial dermis of the skin of neonatal pig by a hollow microneedle. The higher the volume of BSA-FITC solution injected, the higher the amount and the faster the BSA-FITC release rate from skin. These release profiles of BSA-FITC solution were expressed by Fick’s law of diffusion. These results suggest the utilization of hollow microneedle to enhance transdermal delivery of protein and provide useful information for designing an effective hollow microneedle system.
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Authors: J. Kowapradit, Theerasak Rojanarata, Tanasait Ngawhirunpat, A. Apirakaramwong, Warayuth Sajomsang, Praneet Opanasopit
Abstract: In the present study, methylated N-(4-N,N-dimethylaminocinnamyl) chitosan (TM65CM50CS) was synthesized and investigated for oral protein drug delivery by combining with liposomes entrapped bovine serum albumin (FITC-BSA), a model protein. FITC-BSA liposomes composed of egg yolk phosphatidylcholine and sodium oleate in molar ratio of 10:2 were prepared by thin film hydration method. The TM65CM50CS coated liposomal FITC-BSA was evaluated for transport of protein and its cytotoxicity in Caco-2 cells. Moreover, the in vitro stability of BSA in TM65CM50CS coated liposomes was also examined by the degradation of protein from pancreatin. The mean particle size and zeta-potential of liposomes were 101+0.02 nm and -27.44+2.02 mV, respectively. Initial FITC-BSA (2.5% w/w) to lipid showed the highest percentage entrapment efficiency (50.13%) and FITC-BSA content (8.08 mg/g of lipid). The results of FITC-BSA transport showed that TM65CM50CS coated FITC-BSA liposomes enhanced protein permeability across Caco-2 cell monolayers with low cytotoxicity. In addition, these liposomes could protect the degradation of protein from pancreatin. Our studies demonstrated that TM65CM50CS coated liposomes have the potential to be used as an oral protein drug delivery.
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Authors: Tittaya Suksamran, Theerasak Rojanarata, Tanasait Ngawhirunpat, Suwannee Panomsuk, Praneet Opanasopit
Abstract: The aim of this study was to prepare Caalginate and chitosan (CS)Caalginate microparticles for peroral delivery of ovalbumin (OVA). Microparticles containing different loading of OVA (10, 20 and 40 % w/w) were prepared by cross-linking alginate with calcium chloride using an electrohydrodynamic spraying technique, and then coated with CS. The particle sizes of OVA-loaded microparticles were in the range of 1-5 µm. The negative charge was obtained for Caalginate microparticles (-14±1.9 mV) whereas CSCaalginate microparticles were positive charge (+6.06±3.4 mV). Caalginate microparticles with initial 20% w/w OVA showed the highest entrapment efficiency and amount of OVA content (24.91±0.4% and 33.22±0.1 mg/g, respectively) as similar to CSCaalginate microparticles with initial 20% w/w OVA that showed the highest entrapment efficiency and amount of OVA content (35.74±0.1% and 10.35±0.5 mg/g, respectively). It was found that the release rate of OVA from Caalginate microparticles was higher than CSCaalginate microparticles, and the lowest release rate, sustained release for 24 h, was found in the initial 40% w/w OVA. This study revealed that CSCaalginate microparticles have a considerable potential as controlled release antigen delivery systems.
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Authors: Wanlop Weecharangsan, Orapan Paecharoenchai, Nattisa Niyomtham, Praneet Opanasopit, Boon-ek Yingyongnarongkul, Robert J. Lee
Abstract: Polyethylenimine (PEI) was modified by cholic acid at a molar ratio of 1:1. Cholic acid (CA)-modified PEI (PEI-CA) were evaluated for formation of DNA complexes. PEI-CA/pEGFP plasmid DNA complexes were characterized for their size and zeta potential. Gel electrophoresis showed total retardation for PEI-CA/pEGFP complexes formed at weight ratios above 0.25. The particle size and zeta potential of the complexes at a polymer-to-DNA ratio of 0.5 were 295.3 nm and 30.5 mV, respectively. The transfection efficiency of PEI-CA/pEGFP complexes was comparable to unmodified PEI. Cytotoxicity result showed that PEI-CA had lower cytoxicity than PEI. This study suggests that PEI-CA has potential utility as a gene delivery carrier.
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Authors: Nanthida Wonglertnirant, S. Tipwichai, Praneet Opanasopit, Theerasak Rojanarata, Suwannee Panomsuk, Tanasait Ngawhirunpat
Abstract: Ketoprofen transdermal patches (KTPs) were fabricated using an acrylic pressure sensitive adhesive (PSA) polymer. The influence of different factors (amount of PSA, drug content, and pressure applying on the backing membrane during preparation) on the characteristics of ketoprofen patch (thickness, W/A ratio, and adhesiveness of matrix film) and in vitro drug release behavior were investigated. The results revealed the successful fabrication and a good physical appearance of KTPs using acrylic PSA. Microscopic observations, FTIR spectra, and DSC thermograms were permitted to demonstrate that the drug was dispersed molecularly in the polymer. As the amount of PSA in the adhesive matrix was increased, the release rate of ketoprofen was decreased. Contrarily, the drug release rate was increased corresponding to the increase of ketoprofen content in the adhesive matrix. There was no significant difference in the release rate when the pressure applying on the backing membrane was varied. The kinetic of ketoprofen release from acrylic matrix type transdermal patches followed the Higuchis diffusion model.
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Authors: Orapan Paecharoenchai, Tittaya Suksamran, Tanasait Ngawhirunpat, Theerasak Rojanarata, Praneet Opanasopit
Abstract: Chitosan nanoparticles were prepared by crosslinking chitosan (CS) with tripolyphosphate (TPP) solution using electrohydrodynamic spraying technique. The effects of CS and TPP concentration as well as electrical potential on particle size and shape were investigated. Appropriated formulations for preparing nanoparticles were chosen to encapsulate DNA. In vitro evaluation of the obtained nanoparticles as gene carrier such as entrapment efficiency and DNA release was performed. The results showed that 2 mg/ml TPP was dropped at 10 kV into 1 mg/ml CS (MW 20 kDa (F1) and 200 kDa (F2)) yielded the spherical shape and small particles of 227.67 and 240.33 nm, respectively. In DNA entrapment study, all formulations were tested by altering DNA loading to 10, 25 and 50 mg/g of CS. The results revealed that F1 with initial DNA 10 mg/g of CS showed the highest entrapment efficiency of 95.31%. While F2 with initial DNA of 25 mg/g of CS showed the highest entrapment efficiency of 89.16%. The DNA release study from CS nanoparticles indicated that the increasing of DNA amount slowed down the release rate. F1 and F2 with the initial DNA of 10 mg/g of CS had faster release rate than those with 25 and 50 mg/g of CS. It can be concluded that F1 yielded the nanoparticles with the smallest size, high DNA entrapment efficiency and enabled DNA sustained release.
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Authors: Theerasak Rojanarata, Kwanrutai Waewsa-Nga, Parin Buacheen, Praneet Opanasopit, Tanasait Ngawhirunpat
Abstract: An environmentally friendlier, safer and saver method is described for the assay of drugs as hydrochloride salts namely metformin hydrochloride and phenylpropanolamine hydrochloride via the determination of their chloride contents. In this method, an aqueous solution of drug was treated with measured excess of silver nitrate in the presence of nitric acid, followed by the determination of unreacted silver nitrate by Volhard’s method using ammonium thiocyanate titrant and iron(III) alum indicator. To minimize the reagents consumed and wastes generated, the reactions were scaled down to less than 2 mL carried out in microcentrifuge tubes and using micropipettes for the transfer of reagents. The equivalence point was determined by spectrophotometry to diminish visual errors by reading the absorbance of red iron(III) thiocyante complex at 450 nm on microplates, which quickened the measurements of multi-samples. After validation, the method showed satisfactory accuracy and precision and was successfully applied for the assay of both drugs in raw materials, giving the results comparable to the pharmacopeial methods. In addition, the proposed assay was free from the use of harsh, toxic and environmentally harmful chemicals i.e. glacial acetic acid, acetic anhydride, mercuric acetate which are employed in the USP non-aqueous titration methods. Thus, the method is considerably safer for analysts and is a cost-effective and green analytical method suitable for a sustainable environment.
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Authors: Narumon Worachun, Praneet Opanasopit, Theerasak Rojanarata, Tanasait Ngawhirunpat
Abstract: The aim of this study was to prepare microemulsion for transdermal drug delivery of ketoprofen (KP). The physicochemical and chemical properties of microemulsion were evaluated. The microemulsion were composed of isopropyl myristate (IPM) as oil phase, water, PEG40-hydrogenated castor oil (Cremophor® RH40) as surfactant and PEG400 as co-surfactant, and the surfactant: co-surfactant ratio used was 1:1. The viscosity, droplet size, pH, conductivity of microemulsion and skin permeation of KP through shed snake skin were evaluated. The particle size, viscosity and conductivity of microemulsions were in the range of 172-468 nm, 234.82-1067.35 cP and 6.80-20.87µS/cm, respectively. The ratio of IPM and surfactant mixture played an important role on KP loading capacity of microemulsions formulation and skin permeation of KP. While amount of surfactant increased, the loading capacity of KP increased, but the skin permeation of KP decreased. The results suggested that the novel microemulsion system composed of IPM, water, Cremophor® RH40:PEG400 (ratio 1:1) can be applied for using as a transdermal drug delivery carrier.
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