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Gelatin Type A (GA) and sodium alginate (SA) complexes were explored to encapsulate epidermal growth factor (EGF), and thereby to circumvent its proteolytic degradation upon topical application to chronic wounds. Phase diagrams were constructed based on turbidity as a function of GA to SA ratio and pH. Various GA-SA mixtures were compared for polydispersity index, zeta potential, Z-average, and ATR-FTIR spectra. Trypsin digestion and human dermal fibroblast scratch wound assay were done to evaluate the effects of EGF encapsulation. The onset pH values for coacervation and precipitation were closer together in high molecular weight GA (HWGA)-SA reaction mixtures than in low molecular weight GA (LWGA)-SA, which was attributed to strong H-bonding interactions between HWGA and SA probed by ATR-FTIR. EGF incorporation in both HWGA-SA precipitates and LWGA-SA coacervates below the isoelectric point of EGF, but not above it, suggests the contribution of electrostatic interactions between EGF and SA. EGF encapsulated in LWGA-SA coacervates was effectively protected from trypsin digestion and showed better in vitro scratch wound activity compared to free EGF. LWGA-SA coacervates are suggested as a novel delivery system for topical application of EGF to chronic wounds.

Complex coacervates based on natural biopolymers have been explored as a protein delivery system to provide controlled release of loaded protein and circumvent the proteolytic degradation in chronic wounds. The coacervates composed of gelatin A (GA) and sodium alginate (SA) were optimized with respect to turbidity, size, zeta potential, and polydispersity index. Bovine serum albumin (BSA), a model protein, was effectively encapsulated in the coacervates, resulting in protection from trypsin digestion and controlled release. In contrast, EGF was not encapsulated in the same coacervates. Striking difference in the encapsulation efficiencies of BSA and EGF, despite their similar net charges, was attributed to their different levels of binding to GA based on the surface plasmon resonance (SPR) biosensor analysis. In conclusion, GA and SA coacervates can protect the encapsulated protein from proteolytic degradation, demonstrating its potential as a delivery system in the chronic wounds. SPR biosensor is proposed as an analytical tool to study the interactions between polymers and proteins in association with encapsulation efficiency in complex coacervation. The results of EGF studies suggested that GA was not a suitable polymer for EGF encapsulation and therefore, further investigation would be needed to find suitable polymer systems for improved encapsulation efficiency.

Coagulative necrosis often occurs under hypoxic conditions, causing major limitation in the field of tissue engineering especially those dealing with larger tissues and organs. In this study, a comprehensive work has been performed in developing a tailor made design of a dual layered matrix that can produce oxygen to be utilized in tissue engineering application. Optimizations of protocol, ingredient and condition of the system were carried out specifically based on the responses observed from in vitro studies using L6 rat skeletal muscle cell as a candidate. Oxygen was generated from decomposition of encapsulated hydrogen peroxide. Poly (D,L)-lactide-co-glycolide (PLGA) with molecular weights of 90,000 and 110,000 gmol−1 managed to secure good encapsulation of hydrogen peroxide for this application, while the best stirring time during the encapsulation was found to be 8 hours. The PLGA microspheres were coated with a secondary layer of alginate that was pre-grafted with calatase to form the dual layer system. This dual layered architecture has successfully controlled the release rate of oxygen at an optimum level for the survival of muscle cells under hypoxia condition. It was found that muscle cells have low tolerance limit towards the direct contact with hydrogen peroxide, however the cells maintained high viability within encapsulated hydrogen peroxide in the matrix system. It was observed that 4% of encapsulated hydrogen peroxide in the matrix system can produce efficient amount of oxygen at a controlled release manner to sustain the survival of muscle cells under hypoxic condition.

The development of potential applications of biosensors using the sensory systems of vertebrates and invertebrates has progressed rapidly, especially in clinical diagnosis. The biosensor developed here involves the use of Drosophila cells expressing the gustatory receptor Gr5a and an ion-sensitive field-effect transistor (ISFET) sensor device. Gustatory receptor Gr5a is expressed abundantly in gustatory neurons and acts as a primary marker for tastants, especially sugar, in Drosophila. As a result, it could potentially serve as a good candidate for potential biomarkers of diseases in which the current knowledge of the cause and treatment is limited. The developed ISFET was based on the outstanding electrical characteristics of the metal–oxide–semiconductor field-effect transistor (MOSFET) with a subthreshold swing of 85 mV/dec, low leakage current of -12 and high on/off current ratio of 7.3×106. The SiO2 sensing membrane with a pH sensitivity of 34.9 mV/pH and drift rate 1.17 mV/h was sufficient for biosensing applications. In addition, the sensor device also showed significant compatibility with the Drosophila cells expressing Gr5a and their response to sugar, particularly trehalose. Moreover, the interactions between the transfected Drosophila cells and trehalose were consistent and reliable. This suggests that the developed ISFET sensor device could have potential use in the future as a screening device in diagnosis.