Muenduen Phisalaphong

Ultrasound-assisted extraction of capsaicinoids from Capsicum frutescens on a lab-and pilot-plant scale

The influence of operating parameters (solvent type, powder to solvent ratio and temperature) on the ultrasonically assisted extraction of capsaicinoids from dried Capsicum frutescens (fruit) was studied. From the economic perspective, the suitable condition for capsaicinoid extraction by indirect sonication in an ultrasonic bath with a working frequency of 35 kHz was at a ratio of 1g of solid material: 5 ml of 95% (v/v) ethanol, 45 degrees C, where 85% of the capsaicinoids were removed from the raw material in 3h. In an experimental pilot study in 20-l extraction tank at the fixed ultrasonic frequency of 26 kHz and 70 kHz, the recovery of capsaicinoids was 76% and 70%, respectively. It was shown that the ultrasonic extraction produced a significant reduction in extraction time at a lower operational temperature than under a conventional industrial hot maceration process.

Growth of human keratinocytes and fibroblasts on bacterial cellulose film.

Thin films of bacterial cellulose (BC) from a nata de coco culture system were developed, characterized, and investigated for the growth of human keratinocytes and fibroblasts. The average pore diameter and total surface area of the dried BC films estimated by BET were 224 Å and 12.62 m2/g, respectively. With an film thickness of 0.12 mm, the average tensile strength and break strain of the dried films were 5.21 MPa and 3.75%, whereas those of the wet films were 1.56 MPa and 8.00%, respectively. The water absorption capacity of air‐dried film was 5.09 g water/g dried films. For uses in the therapy of skin wounds, the potential biological mechanism of action of BC film was evaluated by using human keratinocytes and fibroblasts. Our results were the first direct demonstration that BC film supported the growth, spreading, and migration of human keratinocytes but not those of human fibroblasts. Expressions of E‐cadherin and the α‐3 chain of laminin confirmed the phenotype of human keratinocytes on BC film.

Biosynthesis and Characterization of Nanocellulose-Gelatin Films

A nanocellulose-gelatin (bacterial cellulose gelatin (BCG)) film was developed by a supplement of gelatin, at a concentration of 1%–10% w/v, in a coconut-water medium under the static cultivation of Acetobacter xylinum. The two polymers exhibited a certain degree of miscibility. The BCG film displayed dense and uniform homogeneous structures. The Fourier transform infrared spectroscopy (FTIR) results demonstrated interactions between the cellulose and gelatin. Incorporation of gelatin into a cellulose nanofiber network resulted in significantly improved optical transparency and water absorption capacity of the films. A significant drop in the mechanical strengths and a decrease in the porosity of the film were observed when the supplement of gelatin was more than 3% (w/v). The BCG films showed no cytotoxicity against Vero cells.

Structural modification and characterization of bacterial cellulose–alginate composite scaffolds for tissue engineering

A novel bacterial cellulose-alginate composite scaffold (N-BCA) was fabricated by freeze drying and subsequent crosslinking with Ca(2+). The N-BCA then underwent a second freeze drying step to remove water without altering the physical structure. A stable structure of N-BCA with open and highly interconnected pores in the range of 90-160 μm was constructed. The N-BCA was stable in both water and PBS. The swelling ability of N-BCA in water was approximately 50 times its weight, which was about 6.5 times that of the freeze dried bacterial cellulose pellicles. N-BCA demonstrated no cytotoxicity against L929 mouse fibroblast cells. For long-term culture, N-BCA supported attachment, spreading, and proliferation of human gingival fibroblast (GF) on the surface. However, under static conditions, the cell migration and growth inside the scaffold were limited. Because of its biocompatibility and open macroporous structure, N-BCA could potentially be used as a scaffold for tissue engineering.

Applicability of Washburn capillary rise for determining contact angles of powders/porous materials.

The Washburn capillary rise (WCR) technique has been widely utilized for determining contact angles of powders or porous materials; however, there are concerns regarding powder size and powder packing, especially for materials that exhibit large contact angle hysteresis. In this paper, some of these concerns were addressed. Due to the large water contact angle hysteresis on flat nylon 6/6 films, these films were ground into powders of different sizes and then used as model packing materials. The powders were packed in glass tubes to result in various packing structures that affected the penetration (i.e. advancing) rate of the test liquids. While all advancing contact angles obtained from WCR were found to be overestimated, more reasonable values were resulted when relatively large powders (e.g. 500-2000 μm) were used to pack the tubes. With larger powders, the packing contained bigger voids and consequently lead to slower penetration rates of the liquids, hence a relatively smaller advancing contact angle. The smaller advancing contact angle obtained from the slower advancing rate was also observed by using the sessile drop method. To verify the applicability of using large powders (500-2000 μm) for contact angle determination by using WCR, the advancing water contact angles of a bacterial cellulose/alginate composite sponge (BCA) with and without UV/ozone treatment were measured. The results showed that by using relatively large powders, WCR could be applied to obtain a reasonable advancing contact angle and assess the wettability change of complex porous materials.

Continuous ethanol production using immobilized yeast cells entrapped in loofa-reinforced alginate carriers

A culture of Saccharomyces cerevisiae M30 entrapped in loofa-reinforced alginate was used for continuous ethanol fermentation in a packed-bed reactor with initial sugar concentrations of 200-248 g/L. Maximum ethanol productivity of 11.5 g/(L·h) was obtained at an ethanol concentration of 57.4 g/L, an initial sugar concentration of 220 g/L and a dilution rate (D) of 0.2 h-1. However, a maximum ethanol concentration of 82.1 g/L (productivity of 9.0 g/(L·h)) was obtained at a D of 0.11 h-1. Ethanol productivity in the continuous culture was 6-8-fold higher than that in the batch culture. Due to the developed carriers high biocompatibility, high porosity, and good mechanical strength, advantages such as cell regeneration, reusability, altered mechanical strength, and high capacity to trap active cells in the reactor were achieved in this study. The immobilized cell reactor was successfully operated for 30 days without any loss in ethanol productivity. The average conversion yield was 0.43-0.45 throughout the entire operation, with an immobilization yield of 47.5%. The final total cell concentration in the reactor was 37.3 g/L (17.7 g/L immobilized cells and 19.6 g/L suspended cells). The concentration of suspended cells in the effluent was 0.8 g/L.

High-temperature ethanol fermentation by immobilized coculture of Kluyveromyces marxianus and Saccharomyces cerevisiae.

Suspended and immobilized cocultures of the thermotolerant yeast, Kluyveromyces marxianus DMKU 3-1042 and the mesophilic flocculent yeast, Saccharomyces cerevisiae M30 were studied for their abilities to improve production and stability of ethanol fermentation. Sugarcane juice and blackstrap molasses, at initial sugar concentrations of 220 g/L, were used as carbon sources. The results indicated that the coculture system could improve ethanol production from both sugarcane juice and blackstrap molasses when the operating temperature ranged between 33 °C and 45 °C. High temperature tolerances were achieved when the coculture was immobilized. The immobilized coculture was more effective in high-temperature ethanol fermentation than the suspended cultures. The coculture immobilized on thin-shell silk cocoon and fermented at 37 °C and 40 °C generated maximal ethanol concentrations of 81.4 and 77.3 g/L, respectively, which were 5.9-8.7% and 16.8-39.0% higher than those of the suspended cultures, respectively.

Effect of molecular weight of chitosan on antimicrobial properties and tissue compatibility of chitosan-impregnated bacterial cellulose films

Bacterial cellulose-chitosan (BC-C) films were developed by immersing purified BC pellicles in 1.5 ~ 2.0% (w/v) acetic acid solutions containing chitosan of varying molecular weights. Effects of different molecular weight of chitosan on physical, biological and antimicrobial properties of the composite films were investigated. The cumulative chitosan absorption capacities with Mw of 141,000, 199,000, and 263,000 were 38.43, 24.65, and 23.89 mg/cm3 of dry BC film, respectively. The cumulative release profiles of chitosan from the films strongly depended on molecular weight of chitosan and pH of solution. The order of release of chitosan from the BC-C films was dependent on molecular weight as follows: Mw 141,000 > Mw 199,000 > Mw 263,000. All BC-C films showed the antimicrobial abilities against Staphylococcus aureus and Aspergillus niger but had no inhibitory effect on the growth of Escherichia coli. The BC-C films supported for adhesion, spreading and proliferation of both human skin keratinocytes and fibroblasts. The antibacterial activity against S. aureus of the BC-C with the highest Mw chitosan (263,000) was higher than those of the others. On the other hand, the BC-C films with the lowest Mw chitosan (141,000) promoted the growth of human skin cells more than those of the others.

Enhanced acetone-butanol production from sugarcane juice by immobilized Clostridium acetobutylicum (ATCC 824) on thin-shell silk cocoons

To promote its performance during acetone-butanol-ethanol (ABE) fermentation, Clostridium acetobutylicum (ATCC 824) was immobilized on a thin-shell silk cocoon (TSC). As a residual from the silk industry, TSC offers a cheap, biocompatible support material. The adsorbed C. acetobutylicum cells digested the TSCs into amino acids as a nitrogen source. It was shown that TSC might promote the phase shift to acetone in the ABE fermentation. At an initial reducing sugar concentration of 90 g/L, the ABE productivity of the immobilized cell culture on TSC (IC-TSC) in batch fermentation was 0.18 g/L/h, and the solvent mixture comprised 6.1 g/L acetone, 15.9 g/L butanol, and 1.9 g/L ethanol. Repeated 4-cycle batch fermentation using IC-TSC significantly improved the ABE productivity. After 48 h of cyclic fermentation, the maximum ABE productivity was 0.43 g/L/h with acetone, butanol and ethanol concentrations of 6.6, 12.9, and 1.1 g/L, respectively.

Biocompatibility and growth of human keratinocytes and fibroblasts on biosynthesized cellulose-chitosan film.

Bacterial cellulose (BC)–chitosan (BCC) films made via bio-co-polymerization by Acetobacter xylinum were developed and characterized for physical and biological properties. With the incorporation of chitosan MW 3 × 104 and 8 × 104 into bacterial cellulose, the modified films (BCC-MW 30 000 and BCC-MW 80 000, respectively) became denser, with a smaller average pore size of 13.1–15.3 nm in dry form. The BCC films have no toxicity against L929 mouse fibroblast cells. Tissue compatibility was then evaluated by growth and spreading of human skin keratinocytes and fibroblasts. The results revealed that the growth of human skin keratinocytes and fibroblasts on the BCC films was comparable to that on the BC film; however, improvement of cell adhesion and spreading on the BCC films was observed in human skin keratinocytes. The results of the biological response experiments showed no significant difference between BCC-MW 30 000 and BCC-MW 80 000.