Pathinan Paengnakorn’s research while affiliated with Chiang Mai University and other places

Biomolecule immobilization on nanomaterials is attractive for biosensors since it enables the capture of a higher concentration of bioreceptor units while also serving as a transduction element. The technique could enhance the accuracy, specificity, and sensitivity of the analytical measurements of biomolecules. However, it was found that the limitation in chemically binding biomolecules on nanoparticle surfaces could only cross-link between the C-terminal and N-terminal. Here, we report the facile one-step synthesis of amine-functionalized silica nanoparticles (AFSNPs). (3-Aminopropyl)triethoxysilane was used as a precursor to modify the functional surface of nanoparticles via the Stöber process. The biomolecules were immobilized to the AFSNPs through itaconic acid, a novel cross-linker that binds between the N-terminal and N-terminal and potentially improves proteins and nucleic acid immobilization onto the nanoparticle surface. The newly developed immobilization approach on AFSNPs for biomolecular detection enhanced the efficiency of ELISA, resulting in increased sensitivity. It might also be easily used to identify different pathogens for clinical diagnostics.

We proposed a specially designed sequential injection (SI) amperometric system coupling with a bioreactor for in-line glucose monitoring in cell culture. The system is composed of three main parts which are the bioreactor, SI system, and electrochemical detection unit. The bioreactor accommodates six individual cell culture units which can be operated separately under different conditions. The SI system enables automatic in-line sampling and in-line sample dilution, with a specially designed mixing unit; therefore, it has the benefits of fast analysis time and less contamination risk. The use of 3D-printed microfluidic components, a mixing channel, and a flow cell helped to reduce operational time and sample volume. A disposable screen-printed electrode (SPE), modified with glucose oxidase (GOD), carbon nanotube, and gold nanoparticle, was used for detection. The developed system provided a linear range up to 3.8 mM glucose in cell culture media. In order to work with cell culture in higher glucose media, the in-line sample dilution can be applied. The developed SI system was demonstrated with mouse fibroblast (L929) cell culture. The results show that glucose concentration obtained from the SI system is comparable with that obtained from the conventional colorimetric method. This work can be further developed and applied for in vitro cell-based experiments in biomedical research.

In the field of dental implants, bone regeneration is needed for a patient with insufficient bone for an implant placement. A mesh membrane is used as a barrier during the augmentation to mechanically stabilise and isolate bone grafts to generate and maintain space for the migration of pluripotential and osteogenic cells. However, the existing membranes, collagen-based and titanium-based, have major problems regarding unfavourable mechanical stability and the need for membrane removal surgery, respectively. Furthermore, these commercial membranes need adjustment to fit the curvature and outline of the bone graft. To overcome such problems, this research was conducted to develop customized biodegradable mesh membranes for dental bone grafts from magnetic resonance imaging or computed tomography using three-dimensional (3D) printing. A biodegradable polycaprolactone (PCL) polymer was used. Various designs of mesh membrane and fabrication procedure were studied. First, a mesh membrane was designed and customized by using a 3D model. Then, an inverse compression mould was made of an acrylic-based resin using LCD-based stereolithography printing technique. Next, the PCL sheet was placed and compressed in the designed mould to create the customized membrane. The results show that the customized membrane prototype exhibited good biocompatibility, biodegradability, and mechanical properties. The results suggest that the developed membrane customization and fabrication procedure could be practically useful for dental bone graft application.

Tissue engineering technology has been advanced and applied to various applications in the past few years. The presence of a bioreactor is one key factor to the successful development of advanced tissue engineering products. In this work, we developed a programmable bioreactor with a controlling program that allowed each component to be automatically operated. Moreover, we developed a new pH sensor for non-contact and real-time pH monitoring. We demonstrated that the prototype bioreactor could facilitate automatic cell culture of L929 cells. It showed that the cell viability was greater than 80% and cell proliferation was enhanced compared to that of the control obtained by a conventional cell culture procedure. This result suggests the possibility of a system that could be potentially useful for medical and industrial applications, including cultured meat, drug testing, etc.

The COVID-19 pandemic has forced analytical chemistry educators in Thailand to change methods of teaching and learning to new normal ones. Higher education has faced additional challenges because of a lack of hands-on experiments and an increasing number of students in foundation chemistry courses being hindered from practicing skills. This work aimed to develop a Lab-at-Home (LAH) for new normal, analytical chemistry experimentation. The LAH implemented a hands-on green chemistry experiment, i.e., colorimetric determination of iron using non-hazardous reagents (supporting Sustainable Development Goal (SDG) 12-responsible consumption and production). The LAH was sent to students at their location before the synchronous class, where the instructors were prompt to supervise. Thus, this supports SDG4-quality education and SDG10-reduced inequalities. The learning outcome achievements, i.e., the analytical characteristics and colorimetry principles comprehension, as well as the ability to perform data analysis, were evaluated by a quiz and laboratory report. LAH satisfaction was assessed by questionnaire and focus group discussion. The learning outcomes were successfully achieved, although students who performed the experiment individually received higher scores than those who did in groups. Students were very satisfied with the LAH as a tool for new normal experimentation, yet some students faced a poor Internet connection during the synchronous online class.

To minimize chemical waste and protect the environment, our team has used green analysis with natural reagents. In this work, we designed a natural-reagent assay kit for iron determination and implemented it in chemistry education in Thailand. The iron assay method was adapted from Thai local wisdom of testing water quality using guava leaves. The guava leaf powder served as a natural reagent in the assay. The kit included equipment, standard and buffer solutions and a manual. A smart device with a built-in camera was used as a detector. Educators in six universities in Thailand implemented the kit in laboratories with modifications depending on their learning outcomes. The kit implementation was evaluated using a survey with questions in four aspects: usability, learning achievement, green chemistry and portability. The high average scores for all questions (> 4.00 of 5.00 points), with the average overall score of 4.53 ± 0.60, indicated satisfaction regarding in all aspects. Using a locally available bio-resource as a natural reagent for green analysis in chemistry education supported sustainable education in Thailand, in terms of quality education (SDG 4) and reduced inequalities (SDG 10) and environmental sustainability (SDG 6—Clean water and sanitation, 12—Responsible consumption and production and 14—Life be-low water).

Silicon photonic devices with either silicon or silicon nitride waveguides have increasingly been used in many applications besides communications, especially as sensors in label-free biosensing, where guided light signals are affected by biorecognition molecules immobilized on the surface. The coating of protein (i.e., bioreceptors) by biochemical process on the waveguide surface is a crucial step in creating a functionalized device that can be used for biosensing. As a conventional method that uses 3-aminopropryltriethoxysilane (APTES) and glutaraldehyde (GA), the APTES-GA method has the limitation of using a GA crosslink, of which the two functional groups can bind to nonspecific proteins, causing irregular binding. In this study, we proposed a new coating technique to avoid such problem by applying APTES silanization with 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide (EDC)-N-hydroxysuccinimide (NHS) protein crosslink, denoted by the APTES-(EDC/NHS) method. The EDC/NHS reaction was shown to be able to immobilize protein in ordered orientation due to consistent arrangement between a carboxylic group of protein molecules and an amine group of covalent-linked APTES on surface. By applying APTES silanization, we circumvented the use of hazardous cleaning agent in the conventional EDC/NHS technique. Several surface characterization techniques were carried out to assess and compare the two biocoating techniques , including scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS), spectroscopic ellipsometry (SE), and atomic force microscopy (AFM). On silicon, the results of antihuman TNF-alpha antibody coating showed that the proposed APTES-(EDC/NHS) technique has better repeatability in terms of less roughness of the coated protein at 1.5 nm compared with 6.3 nm, due to the ordered arrangement of coated antibody molecules. On a silicon nitride waveguide device, the proposed APTES-(EDC/NHS) technique exhibits dense antibody immobilization on a waveguide in SEM images due to stable amide bond formation via EDC/NHS crosslink mechanism. The specificity of the immobilized antibodies was confirmed by enzyme-linked immunosorbent assays (ELISA), with an average optical density at 450 nm of 0.175 ± 0.01 compared with 0.064 ± 0.009 of negative control. The proposed technique also reduced the overall process time since proteins are crosslinked to the silanized waveguide surface in a single step. Citation: Udomsom, S.; Mankong, U.; Paengnakorn, P.; Theera-Umpon, N. Novel Rapid Protein Coating Technique for Silicon Photonic

A simple and cost-effective analytical device based on gas pervaporation and diffusion method for determination of ammonia is proposed. The proposed device is consisted of a well microplate as a donor chamber and a natural indicator-immobilized paper (butterfly pea flower extract) as an acceptor sensor. This allows microliter-scale solution handling. The sample preparation step involved conversion of ammonium ion to ammonia using an alkaline solution, following by gas pervaporation and then diffusion of ammonia across a PTFE membrane into a natural indicator-immobilized paper. The colorimetric change on the paper then was detected by either a smart phone camera or naked eye for quantitative or semi-quantitative analysis, respectively. Under the optimal condition, a linear calibration with a range of 10–100 mg/L ammonium-nitrogen was obtained. The method has a detection limit (3σ) of 4 mg/L ammonium-nitrogen, n = 9. This proposed device was successfully applied for determination of ammonia in swine farming wastewater and chemical fertilizer samples. It could simultaneously perform triplicate run of 18 samples in 15 min (72 samples/hr). The proposed device with pervaporation and diffusion method offers on-site assay and green chemical analysis with relatively high throughput.