Rosa roxburghii tratt callus suspension culture method and application of high-activity rosa roxburghii tratt cell slurry
By optimizing the culture of prickly pear callus using a suspension culture method, the problems of low amplification efficiency, poor uniformity, and easy browning in solid culture were solved. This enabled the production of efficient and uniform prickly pear cell suspension cultures, increased the content of active substances, and expanded the application fields.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- DALIAN POLYTECHNIC UNIVERSITY
- Filing Date
- 2026-02-02
- Publication Date
- 2026-05-15
AI Technical Summary
Existing solid-state culture of prickly pear callus tissue suffers from problems such as low amplification efficiency, poor uniformity, easy browning, and difficulty in scaling up, making it difficult to meet the needs of sustainable high-value utilization.
A stable suspension cell line was established by using the prickly pear callus suspension culture method and by optimizing the induction and subculture medium formulation and culture conditions. Anti-browning agents and inducers were added to the special medium to promote the synthesis of triterpenoids and SOD.
It significantly improved cultivation efficiency and the content of active substances, reduced the browning rate, achieved process uniformity and ease of scale-up, and expanded the application fields.
Smart Images
Figure CN122038264A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of plant tissue culture technology, and more specifically to a method for suspension culture of prickly pear callus and the application of highly active prickly pear cell slurry. Background Technology
[0002] Prickly pear is rich in vitamin C, superoxide dismutase (SOD), triterpenoids, and polyphenols, among other bioactive substances, possessing extremely high nutritional and health benefits. Currently, the acquisition of these bioactive substances mainly relies on fruit harvesting, which is limited by season and geographical location, making it difficult to meet the needs of sustainable production. Among existing technologies, solid-state culture of prickly pear callus tissue is one way to address resource constraints (e.g., CN112352677A). This technology, through optimization of the solid-state culture medium (MS+TDZ+NAA) and culture conditions, successfully induced and cultured prickly pear fruit callus tissue, accumulating certain bioactive substances.
[0003] However, solid-state culture has the following inherent drawbacks: Low amplification efficiency: Callus growth is slow, with a limited fold increase (approximately 1.9 times), making large-scale production difficult. Poor process uniformity: Uneven distribution of nutrients and hormones inside and outside the tissue block leads to inconsistent accumulation of metabolites and poor quality control. Prone to browning and aging: Long-term solid-state culture easily leads to browning of callus tissue and a decline in the ability to synthesize active substances. Difficulty in automation and scaling up: Seamless integration with downstream bioreactor scale-up and automated control is challenging.
[0004] Based on this, a method for the stable and efficient production of highly active prickly pear cell suspension cultures was developed and successfully applied to product development, which is of great significance for realizing the sustainable and high-value utilization of prickly pear resources.
[0005] Therefore, providing a method for suspension culture of prickly pear callus and the application of highly active prickly pear cell plasma is a problem that urgently needs to be solved by those skilled in the art. Summary of the Invention
[0006] In view of this, the present invention provides a method for suspension culture of prickly pear callus and the application of highly active prickly pear cell plasma.
[0007] This invention provides a method for suspension culture of prickly pear callus, which overcomes the shortcomings of existing solid culture technology and obtains a prickly pear cell suspension culture that grows rapidly, is not prone to browning, and has a high content of active substances.
[0008] Another aspect of the present invention provides a highly active prickly pear cell slurry prepared by the above method and its application in functional foods or beverages, cosmetics, and medical dressings.
[0009] To achieve the above objectives, the present invention adopts the following technical solution:
[0010] A method for suspension culture of prickly pear callus includes the following steps: S1. Preparation of starting materials: Select tender leaves of prickly pear seedlings, soak them in 75% ethanol for 30 seconds, disinfect them with 0.1% HgCl2 solution for 8 minutes, rinse them 5 times with sterile water, and cut them into 0.5cm×0.5cm leaf pieces; inoculate the leaf pieces into induction medium, and culture them at 25±1℃, 12h light / 12h dark for 21 days to induce callus tissue, with a light intensity of 2000 lux; screen the pale yellow, loosely structured callus tissue pieces, transfer them to subculture medium, subculture once every 14 days, and purify them for 2 consecutive generations to obtain purified callus tissue.
[0011] The induction medium was based on MS medium, with the addition of 2.0 mg / L 6-BA, 0.5 mg / L NAA (α-naphthaleneacetic acid), 30 g / L sucrose, 7 g / L agar, and pH 5.8.
[0012] The subculture medium was based on MS medium, with the addition of 1.0 mg / L TDZ (Thidiazuron), 0.2 mg / L NAA, 30 g / L sucrose, 7 g / L agar, and pH 5.8.
[0013] S2. Establishment of suspension cell lines: Take 5g of purified callus tissue and inoculate it into a 250mL Erlenmeyer flask containing 100mL of initial liquid culture medium. Incubate at 100rpm, 25±1℃, and in the dark with shaking for 14 days. Filter the culture through a 200-mesh sterile nylon mesh and collect cell clusters of 200-500μm. Transfer the cell clusters to fresh initial liquid culture medium and subculture and screen every 14 days for 3-5 generations to obtain a stable suspension cell line.
[0014] The initial liquid culture medium was based on 1 / 2 MS medium, with the addition of 30 g / L sucrose, 1.0 mg / L TDZ, 0.05 mg / L NAA, and pH 5.8.
[0015] S3. Culture in dedicated culture medium: Inoculate the suspension cell line with dedicated suspension culture medium at a fresh weight of 1-3 g / L, and culture at 110 rpm, 25±1℃, in the dark for 14-20 days.
[0016] The specific suspension medium is based on 1 / 2MS medium, with the addition of 40 g / L sucrose, 2.0 mg / L TDZ, 0.1 mg / L NAA, and 1.5 g / L citric acid (anti-browning agent), pH 5.8 ± 0.1.
[0017] S4. Addition of inducers: On days 10-12 of culture, aseptically add inducers filtered through a 0.22 μm filter membrane, and continue culturing until the end of the cell cycle to obtain the cell system. The inducers are used to specifically promote the synthesis of triterpenoids or SOD.
[0018] The inducer is salicylic acid or methyl jasmonate; the final concentration of salicylic acid is 10-50 μM; the final concentration of methyl jasmonate is 5-20 μM.
[0019] Further, in step S3, the suspension cell line is inoculated at a fresh weight of 1.5 g / L into a special suspension medium and cultured at 110 rpm, 25 ± 1 °C, and in the dark for 16 days. The special suspension medium is based on 1 / 2 MS medium, with the addition of 40 g / L sucrose, 2.0 mg / L TDZ, 0.1 mg / L NAA, 1.5 g / L citric acid, and pH 5.8.
[0020] Furthermore, in step S4, the final concentration of salicylic acid is 30 μM; the final concentration of methyl jasmonic acid is 10 μM.
[0021] Furthermore, a method for preparing a highly active prickly pear cell slurry involves filtering the cell system through a 100-mesh sterile nylon mesh to collect cell clusters; rinsing twice with sterile water, adding an equal volume of sterile water, and homogenizing at 10,000 rpm for 3 minutes to obtain a homogeneous slurry; the slurry has a VC content ≥350 mg / 100 g, a total triterpenoid content ≥15 mg / g, and an SOD activity ≥1000 U / g.
[0022] Furthermore, the method yields a highly active prickly pear cell plasma.
[0023] Furthermore, a highly active prickly pear cell freeze-dried powder is prepared by freeze-drying the aforementioned highly active prickly pear cell slurry at -50°C and 0.1 mbar for 24 hours, with an active substance retention rate of ≥90%.
[0024] Furthermore, a highly active prickly pear cell concentrate is prepared by concentrating the aforementioned highly active prickly pear cell slurry to 1 / 5 of its original volume through rotary evaporation at 50°C and 0.08 MPa, and then adding 0.1% potassium sorbate.
[0025] Furthermore, the application of the highly active prickly pear cell slurry, the highly active prickly pear cell lyophilized powder, or the highly active prickly pear cell concentrate in the preparation of functional foods or beverages involves adding the highly active prickly pear cell slurry, highly active prickly pear cell lyophilized powder, or highly active prickly pear cell concentrate to beverages, yogurt, or capsules at a ratio of 0.5-3%. It is added as a nutritional fortifier to beverages, yogurt, or capsules.
[0026] Furthermore, the application of the highly active prickly pear cell plasma, the highly active prickly pear cell lyophilized powder, or the highly active prickly pear cell concentrate in the preparation of cosmetics involves mixing the highly active prickly pear cell plasma, highly active prickly pear cell lyophilized powder, or highly active prickly pear cell concentrate with glycerin, sodium hyaluronate, or an emulsifier at a ratio of 3-8%. This mixture is then combined with an emulsion system to prepare skincare products with antioxidant and whitening functions.
[0027] Furthermore, the application of the highly active prickly pear cell plasma, the highly active prickly pear cell lyophilized powder, or the highly active prickly pear cell concentrate in the preparation of medical dressings involves mixing the highly active prickly pear cell plasma, the highly active prickly pear cell lyophilized powder, or the highly active prickly pear cell concentrate with bacterial cellulose at a mass ratio of 1:5, lyophilizing it into a 0.5 mm thick film, and sterilizing it with ethylene oxide to prepare an antibacterial dressing. This dressing can then be combined with other biomaterials such as bacterial cellulose to prepare dressings that promote wound healing.
[0028] As can be seen from the above technical solution, compared with the prior art, the present invention discloses a method for suspension culture of prickly pear callus and the application of highly active prickly pear cell plasma, which has the following beneficial effects: (1) The culture efficiency was significantly improved: the biomass of the suspension culture increased by 2.6 times, which was significantly higher than that of the solid culture in CN112352677A by 1.9 times.
[0029] (2) The content of active substances is greatly improved: The vitamin C content in the cell plasma obtained by this invention can reach 398 mg / 100g, and the total triterpenoid content can reach 22.7 mg / g under the action of the inducer, which are significantly better than the solid culture in CN112352677A (vitamin C 45.12 mg / 100g, total triterpenoid 8.45 mg / g).
[0030] (3) Effectively solve the browning problem: By using special culture medium and anti-browning agent, the browning rate can be controlled below 10%.
[0031] (4) The process is uniform and controllable, and easy to scale up: The liquid system has good uniformity, which lays the foundation for subsequent large-scale cultivation in bioreactors.
[0032] (5) Diversified application pathways: It has opened up new applications of prickly pear callus tissue in medical materials and other fields. Attached Figure Description
[0033] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.
[0034] Figure 1 The image shows a prickly pear suspension cell cluster prepared in Example 1, which shows that the cell clusters are uniform in size and have a loose structure.
[0035] Figure 2 This is a comparison chart of the biomass (dry weight) growth curves of Example 1 (suspension culture) and Comparative Example 1 (solid culture) during the culture period.
[0036] Figure 3 The bar chart shows the comparison of vitamin C and total triterpenoid content between the products obtained in Comparative Example 1 and Examples 1-3. Detailed Implementation
[0037] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0038] Example 1: Establishment and culture of prickly pear suspension cell line The method for inducing and subculturing callus from prickly pear leaves includes the following steps: (1) Preparation of starting material: Select tender leaves of prickly pear seedlings, soak them in 75% ethanol for 30s, disinfect them with 0.1% HgCl2 solution for 8min, rinse them with sterile water 5 times, and cut them into 0.5cm×0.5cm leaf pieces; inoculate the leaf pieces into induction medium, and culture them at 25±1℃, 12h light / 12h dark for 21 days to induce callus tissue, with a light intensity of 2000 lux; screen the pale yellow, loosely structured callus tissue pieces, transfer them into subculture medium, subculture once every 14 days, and purify them for 2 consecutive generations to obtain purified callus tissue.
[0039] The induction medium was based on MS medium, with the addition of 2.0 mg / L 6-BA, 0.5 mg / L NAA, 30 g / L sucrose, and 7 g / L agar, pH 5.8.
[0040] The subculture medium was based on MS medium, with the addition of 1.0 mg / L TDZ, 0.2 mg / L NAA, 30 g / L sucrose, 7 g / L agar, and pH 5.8.
[0041] (2) Construction of the prickly pear cell suspension culture system Establishment of suspension cell lines: Take 5g of purified callus tissue and inoculate it into a 250mL Erlenmeyer flask containing 100mL of initial liquid culture medium. Incubate at 100rpm, 25±1℃, and in the dark with shaking for 14 days. Filter the culture through a 200-mesh sterile nylon mesh to collect cell clusters of 200-500μm. Transfer the cell clusters to fresh initial liquid culture medium and subculture and screen every 14 days for 3 consecutive generations to obtain a stable suspension cell line.
[0042] The initial liquid culture medium was based on 1 / 2 MS medium, with the addition of 30 g / L sucrose, 1.0 mg / L TDZ, 0.05 mg / L NAA, and pH 5.8.
[0043] Culture in a special culture medium: Inoculate the suspension cell line with a fresh weight of 1.5 g / L into a special suspension culture medium and culture at 110 rpm, 25°C and in the dark for 16 days.
[0044] The dedicated suspension medium was based on 1 / 2 MS medium, with the addition of 40 g / L sucrose, 2.0 mg / L TDZ, 0.1 mg / L NAA, 1.5 g / L citric acid, and pH 5.8.
[0045] Cell growth status see Figure 1 .
[0046] After cultivation, the biomass dry weight multiplication factor was 2.6, the vitamin C content was 398 mg / 100g, the total triterpenoid content was 16.8 mg / g, the SOD enzyme activity was 1247 U / g, and the browning rate was 8%.
[0047] The formula for calculating biomass is: Biomass = Number of cells harvested after culture - Number of cells at the time of inoculation.
[0048] Biomass dry weight multiplication factor = dry weight of cells harvested after culture / initial dry weight of cells at the time of inoculation.
[0049] Example 2: Promotion of active substance synthesis by inducible salicylic acid Based on Example 1, on day 12 of culture, sterile filtered salicylic acid stock solution was added to the experimental group culture system to bring the final concentration to 30 μM. Cells were harvested after another 4 days of culture. Analysis showed that, compared to Example 1, the total triterpenoid content in the experimental group cells increased to 22.7 mg / g, an increase of 35%, and the SOD enzyme activity was 2250 U / g.
[0050] Example 3: Promotion of active substance synthesis by the inducer methyl jasmonate Based on Example 1, on day 12 of culture, sterile filtered methyl jasmonate stock solution was added to the experimental group culture system to bring the final concentration to 10 μM. Cells were harvested after another 4 days of culture. Analysis showed that, compared to Example 1, the total triterpenoid content in the experimental group cells increased to 20.2 mg / g, an increase of 20%, and the SOD enzyme activity was 1426 U / g.
[0051] Comparative Example 1 (Traditional Solid Culture) Following the method described in Example 2 of patent CN112352677A, homologous prickly pear callus was inoculated onto optimal solid culture medium (MS + 3.0 mg / L TDZ + 0.1 mg / L NAA + 60 g / L sucrose + 5 g / L agar, pH 5.8) and cultured statically for 28 days at 25°C under 12 hours of light per day. After culture, the biomass dry weight proliferation rate was measured to be 1.9, the vitamin C content was 45.12 mg / 100g, and the total triterpenoid content was 8.45 mg / g.
[0052] Experimental Example 1: Preparation and Key Indicator Analysis of Highly Active Cell Plasma (1) Cell harvesting and pretreatment: Take a total of 500 mL of the cell culture system after the culture in Examples 1-3, filter it through a 100-mesh sterile nylon mesh, and collect the cell clusters on the filter. Rinse the cell clusters twice with pre-cooled sterile deionized water (4°C), each time using approximately twice the volume of the cell clusters, to thoroughly remove any residual culture medium components.
[0053] Take a total of 5 g of cells after the culture of Comparative Example 1, and rinse the cell cluster twice with pre-cooled sterile deionized water (4℃) to thoroughly wash away residual culture medium components.
[0054] (2) Homogenization to prepare slurry: Weigh the rinsed wet cell clumps and record the fresh weight. Add an equal volume of pre-cooled sterile deionized water (4°C) to the cell clumps at a ratio of 1:1 (w / v). Transfer the mixture to a pre-cooled glass homogenizer and homogenize mechanically under ice bath conditions. Homogenization conditions: 10,000 rpm, intermittent homogenization (30 s working, 15 s pause), total time 3 minutes, until a homogeneous, milky white slurry without obvious particles is obtained.
[0055] (3) Slurry quality control and analysis: The prepared slurry was centrifuged at 4℃ and 10,000 rpm for 10 minutes, and the supernatant was used for the determination of key active ingredients. Simultaneously, a portion of the homogenized but not centrifuged whole slurry was used for the detection of key indicators. The results are as follows: Appearance and physicochemical properties: The slurries of Examples 1-3 are all milky white homogeneous liquids, while the slurry of Comparative Example 1 is a pale yellow homogeneous liquid; the slurries of Examples 1-3 and Comparative Example 1 all have a slightly characteristic aroma of prickly pear, and the pH value is 5.5±0.2.
[0056] Vitamin C (VC) content: determined by 2,6-dichlorophenolindophenol titration method, the slurry meets the requirement of VC content ≥35 mg / 100g.
[0057] Total triterpenoid content: determined by vanillin-glacial acetic acid colorimetric method (using oleanolic acid as standard), the total triterpenoid content in the slurry meets the threshold of ≥15 mg / g.
[0058] Superoxide dismutase (SOD) activity: determined by nitroblue tetrazolium (NBT) photochemical reduction method, the SOD activity in the slurry meets the requirement of ≥1000 U / g (based on fresh weight).
[0059] (4) Preservation of slurry: If the prepared highly active prickly pear cell plasma is not used immediately, it can be dispensed into sterile brown vials, filled with nitrogen, sealed, and stored at -80℃, which can effectively maintain the active ingredients for more than 6 months.
[0060] The key indicators of Examples 1-3 were compared with those of Comparative Example 1, and the results are shown in Table 1.
[0061] Table 1
[0062] As shown in Table 1, the suspension culture method of the present invention is significantly superior to the traditional solid culture technology in terms of culture efficiency (43% shorter cycle and 37% higher proliferation rate), content of active substances (8.8 times higher VC and 1.99-2.69 times higher total triterpenes) and browning control (75% lower browning rate).
[0063] Figure 2 This is a comparison chart of the biomass (dry weight) growth curves of Example 1 (suspension culture) and Comparative Example 1 (solid culture) during the culture period. Figure 3 The bar chart shows the comparison of vitamin C and total triterpenoid content between the products obtained in Comparative Example 1 and Examples 1-3.
[0064] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A method for suspension culture of prickly pear callus, characterized in that, Includes the following steps: S1. Preparation of starting materials: Select tender leaves of prickly pear seedlings, soak them in 75% ethanol for 30 seconds, disinfect them with 0.1% HgCl2 solution for 8 minutes, rinse them 5 times with sterile water, and cut them into 0.5cm×0.5cm leaf pieces; inoculate the leaf pieces into induction medium, and culture them at 25±1℃, 12h light / 12h dark for 21 days to induce callus tissue, with a light intensity of 2000 lux; screen the pale yellow, loosely structured callus tissue pieces, transfer them to subculture medium, subculture once every 14 days, and purify them for 2 consecutive generations to obtain purified callus tissue; The induction medium was based on MS medium, with the addition of 2.0 mg / L 6-BA, 0.5 mg / L NAA, 30 g / L sucrose, 7 g / L agar, and pH 5.
8. The subculture medium was based on MS medium, with the addition of 1.0 mg / L TDZ, 0.2 mg / L NAA, 30 g / L sucrose, 7 g / L agar, and pH 5.
8. S2. Establishment of suspension cell lines: Take 5g of purified callus tissue and inoculate it into a 250mL Erlenmeyer flask containing 100mL of initial liquid culture medium. Incubate at 100rpm, 25±1℃, and in the dark with shaking for 14 days. Filter the culture through a 200-mesh sterile nylon mesh and collect cell clusters of 200-500μm. Transfer the cell clusters to fresh initial liquid culture medium and subculture and screen every 14 days for 3-5 generations to obtain a stable suspension cell line. The initial liquid culture medium was based on 1 / 2 MS medium, with the addition of 30 g / L sucrose, 1.0 mg / L TDZ, 0.05 mg / L NAA, and pH 5.
8. S3. Culture in dedicated culture medium: Inoculate the suspension cell line with dedicated suspension culture medium at a fresh weight of 1-3 g / L, and culture at 110 rpm, 25±1℃, in the dark for 14-20 days. The specific suspension medium is based on 1 / 2 MS medium, with the addition of 40 g / L sucrose, 2.0 mg / L TDZ, 0.1 mg / L NAA, and 1.5 g / L citric acid, pH 5.8 ± 0.1; S4. and / or addition of inducers: On days 10-12 of culture, aseptically add inducers filtered through a 0.22μm filter membrane, and continue culturing until the end of the cycle to obtain the cell system; The inducer is salicylic acid or methyl jasmonate; the final concentration of salicylic acid is 10-50 μM; the final concentration of methyl jasmonate is 5-20 μM.
2. The method for suspension culture of prickly pear callus according to claim 1, characterized in that, In step S3, the suspension cell line was inoculated at a fresh weight of 1.5 g / L into a special suspension medium and cultured at 110 rpm, 25 ± 1 °C, and in the dark for 16 days. The special suspension medium was based on 1 / 2 MS medium, with the addition of 40 g / L sucrose, 2.0 mg / L TDZ, 0.1 mg / L NAA, 1.5 g / L citric acid, and pH 5.
8.
3. The method for suspension culture of prickly pear callus according to claim 1, characterized in that, In step S4, the final concentration of salicylic acid is 30 μM; the final concentration of methyl jasmonate is 10 μM.
4. A method for preparing highly active prickly pear cell plasma, characterized in that, The cell system described in any one of claims 1-3 is filtered through a 100-mesh sterile nylon mesh to collect cell clusters; after rinsing twice with sterile water, an equal volume of sterile water is added, and the mixture is homogenized at 10,000 rpm for 3 minutes to obtain a homogeneous slurry; the slurry has a VC content ≥350mg / 100g, a total triterpenoid content ≥15mg / g, and an SOD activity ≥1000U / g.
5. The highly active prickly pear cell plasma prepared by the method of claim 4.
6. A highly active lyophilized powder of prickly pear cells, characterized in that, The highly active prickly pear cell plasma as described in claim 5 is prepared by freeze-drying at -50°C and 0.1 mbar for 24 hours, with an active substance retention rate of ≥90%.
7. A highly active prickly pear cell concentrate, characterized in that, It is prepared by concentrating the highly active prickly pear cell plasma as described in claim 5 to 1 / 5 of its original volume through rotary evaporation at 50°C and 0.08 MPa, and then adding 0.1% potassium sorbate.
8. The application of the highly active prickly pear cell slurry of claim 5, the highly active prickly pear cell lyophilized powder of claim 6, or the highly active prickly pear cell concentrate of claim 7 in the preparation of functional foods or beverages, characterized in that, Add highly active prickly pear cell plasma, highly active prickly pear cell lyophilized powder, or highly active prickly pear cell concentrate to beverages, yogurt, or capsules at a ratio of 0.5-3%.
9. The application of the highly active prickly pear cell plasma of claim 5, the highly active prickly pear cell lyophilized powder of claim 6, or the highly active prickly pear cell concentrate of claim 7 in the preparation of cosmetics, characterized in that, Mix highly active prickly pear cell plasma, highly active prickly pear cell lyophilized powder, or highly active prickly pear cell concentrate with glycerol, sodium hyaluronate, or emulsifier at a ratio of 3-8%.
10. The application of the highly active prickly pear cell plasma of claim 5, the highly active prickly pear cell lyophilized powder of claim 6, or the highly active prickly pear cell concentrate of claim 7 in the preparation of medical dressings, characterized in that, Highly active prickly pear cell plasma, highly active prickly pear cell freeze-dried powder, or highly active prickly pear cell concentrate are mixed with bacterial cellulose at a mass ratio of 1:5, freeze-dried into a 0.5 mm thick film, and sterilized with ethylene oxide to produce an antibacterial dressing.