TWI573596B - Ganoderma lucidum polysaccharide bio-wound dressing - Google Patents

Description

Ganoderma lucidum polysaccharide biological dressing

Use of bacterial cellulose with high mechanical properties and texture, chitosan with accelerated wound healing and good antibacterial ability, and sodium alginate with good water absorption capacity, through sol casting and cross-linking technology to form dry recombinant biological dressing The substrate is then adsorbed to extract the Ganoderma lucidum polysaccharide biological dressing, and the microwave sterilization and microwave freeze drying process are used to rapidly prepare the Ganoderma lucidum polysaccharide biological dressing. Through the mouse animal experiment, Ganoderma lucidum polysaccharide biological dressing can promote the wound healing effect of normal and STZ-induced diabetic mice.

The most commonly used raw materials for biological dressing development research are chitosan (Jayakumar et al., 2011) and bacterial cellulose (Czaja et al., 2006). Chitosan has immunostimulatory effects, can attract polymorphic leukocyte aggregation, activate macrophages, promote fibroblast growth, stimulate cytokine production, accelerate wound healing, inhibit Pseudomonas aeruginosa and Proteus (Burkatovskaya et al. , 2006). Bacterial cellulose has good biocompatibility and network structure, and can replace synthetic polymers as biomedical materials (Fernandes et al., 2010; Chiaoprakobkij et al., 2011), and can also improve dressing water retention and reduce wound water. Vapor evaporation rate, developed a composite biological dressing that promotes rapid wound healing (Yang, 2011).

Ganoderma lucidum fruiting body is also a good source of chitin, made by alkali treatment Butan can be used in wound therapy to promote wound healing in rats (Su et al., 1997). However, as disclosed in Patent Publication No. 442496, the Ganoderma lucidum fruit body or its residue, which needs to be cultivated for 5 to 6 months, is immersed in an organic solvent, dried, and then digested with sodium hydroxide, dechlorinated with hypochlorite, and washed to obtain polydip. Amine fiber is used as a material for wound healing, but it needs to be treated with chemicals, which will cause environmental pollution.

In addition, Ganoderma lucidum polysaccharides can resist tumors, increase immunity, resist oxidation, reduce fat oxidation, and improve hyperglycemia (Boh et al., 2003; Gao et al., 2004; Chen et al., 2009; Jia et al., 2009). Functionality of anti-glycation end products (Meng et al., 2011; Rao et al., 2012; Hori et al., 2012). Ganoderma lucidum polysaccharides are applied to wound healing treatment by spraying to accelerate wound healing in rats (Zhan, 2005).

According to the I303557 invention patent of Chen Hezhuang (2008), using cereals or miscellaneous grains as the substrate for solid-state fermentation of Ganoderma lucidum, the fermentation product of Ganoderma lucidum mycelium can be obtained within one month, and the method can be utilized for whole grain fermentation, and then Hot water extraction of Ganoderma lucidum polysaccharide can solve the problems of stability, safety, hygiene, pollution reduction and rapid production of Ganoderma lucidum polysaccharide raw materials. The time required to extract the polysaccharide from the microwave hot water is only 10 minutes shorter than the conventional hot water reflux extraction and the ultrasonic extraction for 20 minutes and the efficiency is good (Chen and Chen, 2013).

The sterilization methods commonly used in wound dressings can be roughly divided into physical and chemical methods, and the purpose is to make microorganisms inactivated and unable to survive. The physical sterilization method can be classified into a heating method, a radiation irradiation method, and a novel microwave sterilization according to classification. In the chemical sterilization method, a bactericide is applied to cause the cell membrane of the cell to cause the protein and DNA to replicate. Among them, the sterilization of the sterilization kettle is caused by the poor heat transfer effect of the solid matter, so the temperature is high for a long time and the quality of the product is poor. The use of radiation exposure is more common with gamma-rays, but this requires special protection against radiation leakage facilities. To be outsourced, it takes time and cost and scheduling. Microwave sterilization is now widely used and relevant The equipment is also quite mature, such as food industry, denture industry and medical waste (Huang & Sites, 2007; Tounci et al., 2008; Dovigo et al., 2009; Kim et al., 2012). application.

Drying is an extremely important part of the dressing process. Because the dry dressing can absorb the secretions, it can also avoid the problem of growth of the biological dressings in the storage microorganisms, thereby reducing the weight and volume, facilitating packaging and storage, and facilitating transportation. And extend the shelf life. The use of microwave as a heat source for the freeze-drying process provides ice crystal sublimation, which overcomes the traditional heat conduction resistance, so it can greatly shorten the freeze-drying time and maintain good quality. Huang (2012) uses microwave freeze-drying to treat the body of A. sinensis, and traditional freezing Compared with drying, the drying time can be shortened by 34 hours, and the energy consumption can be greatly reduced by more than 90%. The appearance of the finished product after drying is not significantly different from the conventional freeze-drying.

The bacterial cellulose, chitosan and sodium alginate are uniformly homogenized and stirred, and then smoothly applied to the glass mold to dry the surface, and then the whole piece of the recombinant dressing is immersed in the calcium chloride solution to crosslink, and the mold is removed. The wet recombinant biofilm is freeze-dried or microwave freeze-dried to form a dried biological dressing, and then adsorbs the polysaccharide extract of the solid extract of Ganoderma lucidum which has been extracted by microwave, and then sterilized by microwave, then freeze-dried or microwave freeze-dried. The Ganoderma lucidum polysaccharide biological dressing is packaged and the Ganoderma lucidum polysaccharide biological dressing can promote wound healing.

Figure 1 is a flow chart of the manufacture of the ganoderma lucidum polysaccharide biological dressing of the present invention.

Figure 2 shows the temperature rise curve of 50 mL of Ganoderma lucidum polysaccharides (200-400W).

Figure 3 shows the extraction rate of Ganoderma lucidum polysaccharides extracted with different microwave power (200-400W) for 10 min.

Figure 4 shows the heating curve of 25g Ganoderma lucidum biological dressing treated with different microwave power.

Figure 5 shows the effect verification of 300W microwave sterilization Ganoderma lucidum biological dressing.

Figure 6 is a 300W microwave sterilization biological dressing for E. coli and Staphylococcus aureus mortality

Figure 7 shows the drying curve and temperature curve of 25g Ganoderma lucidum biological dressing treated with different microwave freeze-drying power.

Figure 8 is the wound healing effect of Ganoderma lucidum polysaccharide biological dressing on mice.

The ganoderma lucidum polysaccharide biological dressing manufacturing method firstly homogenizes the bacterial cellulose, the chitosan and the sodium alginate with water, and the water content is more than 90%, and is smoothly applied to the glass mold at 30 ° C and relative humidity 40. The surface was dried in a constant temperature and humidity incubator for 2 hours, and then the whole piece of the recombinant dressing was immersed in a 3.5% calcium chloride solution for crosslinking for 2 hours. The mold was removed as a wet recombinant biofilm, and then dried by 50W microwave drying. Become a dry biological dressing.

Ganoderma lucidum solid-state fermented grain milling powder was uniformly mixed into water 50mL solution with a solid-liquid ratio of 1:20, heated by 200~400W microwave extraction (Fig. 2), and the Ganoderma lucidum polysaccharide solution was quickly extracted for 10 minutes. c) adsorbing the Ganoderma lucidum polysaccharide extract with the above dried biological dressing, and then sterilizing the 25 g of the wet Ganoderma lucidum polysaccharide biological dressing with microwaves of different powers of 100, 200 and 300 W, and heating the temperature (Fig. 4) 300W microwave heating is the fastest heating, so the Ganoderma lucidum polysaccharide biological dressing is sterilized in 300W microwave heating for 40, 45 and 50 seconds, and it is flattened on PDA. After culturing on the plate of the nutrient, it was completely sterilized by microwave sterilization at 300W for 50 seconds, so there was no microbial growth (Fig. 5), and the bactericidal degree of 6D was reached for the indicator bacteria Escherichia coli and Staphylococcus aureus (Figure 6). ). However, the microwave sterilization treatment of the Ganoderma lucidum polysaccharide biological dressing, and microwave drying, wherein 50W microwave freezing takes about 1 hour to complete the drying, and the Ganoderma lucidum polysaccharide biological dressing product temperature is lower than 30 ° C (Figure 7), Therefore, low temperature drying can maintain good quality. Then, using Ganoderma lucidum polysaccharide biological dressing can avoid excessive infiltration of normal mice and STZ-induced diabetic mice, and at the same time, under the action of Ganoderma lucidum polysaccharide-stimulated cells, the skin wound recovery on the second day of treatment is significantly better than that of the control group treated with gauze. Shorten wound healing time (Figure 8).

Claims (9)

A method for producing a ganoderma lucidum polysaccharide biological dressing, comprising: (a) extracting a ganoderma lucidum polysaccharide extract of a solid fermentation product of Ganoderma lucidum by microwave, adsorbing it in (b) a recombinant biological dressing, and then using (c) microwave sterilization and (d) Microwave freeze-drying to make a Ganoderma lucidum polysaccharide biological dressing. The manufacturing method according to claim 1, wherein (a) the microwave extraction of the ganoderma lucidum polysaccharide extract of the solid fermentation product of the ganoderma lucidum is first inoculated with the ganoderma lucidum in the solid substrate of the grain, wherein the moisture content of the whole grain substrate is controlled at 30 to 60. %, about 1 to 5 weeks at 25~35 °C, so that the ganoderma lucidum mycelium grows over the substrate, then sterilize, dry, and grind; finally add water to the ratio of 1:5~1:20 solid-liquid, microwave The heat source can be extracted for about 5 to 30 minutes to obtain the polysaccharide extract of Ganoderma lucidum. The manufacturing method according to claim 1, wherein (b) the recombinant biological dressing is controlled by homogenizing the bacterial cellulose, chitosan and sodium alginate in a ratio of 2:1:1, and then controlling the moisture content. More than 90%, smooth coating on the glass mold, surface drying in a constant temperature and humidity incubator for 2 hours, and then immersing the entire piece of the recombinant dressing in 3.5% calcium chloride solution for 2 hours, the mold is wet The recombinant biological dressing, after freeze-drying, can adsorb the ganoderma lucidum polysaccharide extract of step (a), which is a semi-finished product of the ganoderma lucidum polysaccharide biological dressing. The manufacturing method according to claim 1, wherein (c) microwave sterilization is a semi-finished product of a ganoderma lucidum polysaccharide biological dressing, wherein the microwave is used as a heat source, and the temperature of the ganoderma lucidum polysaccharide biological dressing is heated to 90 ° C for several seconds to achieve sterilization. effect. The manufacturing method according to claim 1, wherein (d) microwave freeze-drying, the sterilized ganoderma lucidum polysaccharide biological dressing needs to be frozen first, and then sterilized by microwave as a heat source, so that the dried ganoderma lucidum polysaccharide biological dressing has low moisture content. At 5%. A Ganoderma lucidum polysaccharide biological dressing prepared by the method of any one of claims 1 to 5. Ganoderma lucidum polysaccharide biological dressing as claimed in claim 6 for use in wound healing. A use of the ganoderma lucidum polysaccharide biological dressing of claim 6 for preparing a dressing for promoting wound healing. The use according to claim 8, wherein the ganoderma lucidum polysaccharide biological dressing can reduce the wound in two days for normal and diabetic wounds, thereby shortening the wound healing time.