TWI712415B - Topical composition, medicinal composition containing inactivated culture and use thereof for facilitating wound healing and scar reduction - Google Patents

Abstract

The present invention relates to a topical composition of facilitating wound healing and reducing scars, which includes an inactivated culture of Lactobacillus species as an effective ingredient, thereby producing a medical composition of facilitating wound healing and reducing scars.

Description

A topical composition containing a dead bacteria culture of Lactobacillus, a medical composition and its use in promoting wound healing and reducing scars

The present invention relates to a topical composition, in particular to a topical composition and a medical composition containing a dead Lactobacillus culture and its use in promoting wound healing and reducing scars.

The wound healing process is generally divided into four stages. The first stage hemostasis (hemostasis), that is, the wound occurs until hemostasis. The second stage is the inflammatory phase. The body's immune system is designed to resist the invasion of foreign bacteria and microorganisms, and the wounds appear to be slightly red, swollen, hot, and painful.

The third stage is the proliferative phase. New capillaries will gradually grow at the wound site, bringing more collagen tissue to fill the wound. This stage also includes the proliferation of granulation tissue. In the process, fibroblasts are activated to express α-smooth muscle actin (α-SMA) and transform into myofibroblasts. Cell (myofibroblast). Myofibroblasts have the ability to restructure the extracellular matrix (ECM) material and shrink the wound, so that the wound will gradually close.

The fourth stage is the maturity or remodeling phase. After the wound is closed and healed, the excess capillaries degenerate and shrink. The collagen tissue is elongated and arranged neatly, and the scar becomes flat and light.

In the process of wound healing, the conversion of fibroblasts into myofibroblasts is an important step in wound healing; however, during the remodeling phase, if myofibroblasts are over-activated, improper scars and fibrosis will occur ( fibrosis), leading to scars. Therefore, to avoid incomplete wound healing or scarring, the key is to control myofibroblasts so that they are not over-activated. It is currently known that TGF-β is an important factor for activating myofibroblasts. How to properly regulate the performance of TGF-β is one of the key points for promoting wound healing and reducing scars.

Probiotics have been developed for a long time, and probiotics are safe and have no side effects. Past studies have found that using L.plantarum live bacteria locally to wipe burned mouse wounds can increase the phagocytic ability of local immune cells and reduce the number of pathogenic bacteria. Increase tissue repair capabilities. Further research has also found that topical administration of live Lactobacillus plantarum can reduce the number of wound bacteria in clinical burn patients and accelerate wound healing. Screening produce high amounts of exopolysaccharide (EPS) of the viable Lactobacillus (L. plantarum) or Lactobacillus brevis (L. brevis) viable culture medium can promote wound healing in rats. However, without empirical research, it is impossible to predict whether different types of probiotics have the ability to promote wound healing.

In view of this, it is necessary to provide a topical composition containing probiotics to enhance the application of probiotics in wound dressings.

Therefore, one aspect of the present invention is to provide a topical composition for promoting wound healing and reducing scars, which contains a dead Lactobacillus culture as an effective ingredient.

Another aspect of the present invention is to provide a medical composition for promoting wound healing and reducing scars, which contains a dead Lactobacillus culture as an active ingredient.

Another aspect of the present invention is to provide a dead bacteria culture of Lactobacillus for the manufacture of a pharmaceutical composition that promotes wound healing and reduces scars, wherein the dead Lactobacillus culture is derived from Lactobacillus paracasei ( L. paracasei ) GMNL-653, and the dead Lactobacillus culture contains pure dead bacteria and/or dried bacteria powder to promote wound healing and reduce scars.

According to the above aspect of the present invention, a medical composition for promoting wound healing and reducing scars is proposed, which contains a dead Lactobacillus culture as an active ingredient. In this embodiment, the dead Lactobacillus culture may be derived from Lactobacillus paracasei ( Lactobacillus paracasei ) GMNL-653, and Lactobacillus paracasei GMNL-653 was deposited at Hsinchu Foods, Taiwan on February 26, 2016. Biological Resources Conservation and Research Center (BCRC), Food Industry Development Research Institute, No. 331 Road, the deposit number is BCRC 910721.

In an embodiment of the present invention, the aforementioned dead bacteria culture of Lactobacillus comprises a pure dead bacteria culture solution and/or dry bacteria powder.

According to another aspect of the present invention, a medical composition for promoting wound healing and reducing scars is provided, which contains a dead Lactobacillus culture as an active ingredient.

In an embodiment of the present invention, the effective dose of the above-mentioned dead Lactobacillus culture may be, for example, 1×10 ⁹ bacterial cells/mL to 1×10 ¹⁰ bacterial cells/mL.

In an embodiment of the present invention, the dosage form of the above-mentioned pharmaceutical composition may be, for example, a gel, a gel-like dressing, a sponge-like dressing, a film-like dressing, or any combination of the foregoing.

According to another aspect of the present invention, a dead bacteria culture of Lactobacillus is proposed for the manufacture of a pharmaceutical composition that promotes wound healing and reduces scars. The dead Lactobacillus culture can be derived from, for example, side cheese milk. L. paracasei GMNL-653 (Accession Number: BCRC 910721), and the dead Lactobacillus culture contains pure dead bacteria and/or dry bacteria powder.

Application of the local composition and medical composition for promoting wound healing and reducing scars of the present invention, which contains dead Lactobacillus culture as an effective ingredient, can significantly promote wound healing and reduce scars, and then be used for manufacturing to promote wound healing and reduce scars The purpose of the medical composition.

In order to make the above and other objects, features, advantages and embodiments of the present invention more comprehensible, the detailed description of the attached drawings is as follows: [Figure 1A] shows the human skin HS68 fibroblasts according to an embodiment of the present invention A bar graph showing the relative expression of type 1 collagen after co-cultivation of different types of dead lactic acid bacteria strains.

[Figure 1B] is a bar graph showing the relative expression levels of type 1 matrix metalloproteinases after co-cultivation of a human skin HS68 fibroblast cell line with different concentrations of dead lactic acid bacteria strains according to an embodiment of the present invention.

[Fig. 2A] to [Fig. 2B] show the wound appearance (Fig. 2A) and wound size curve diagram (Fig. 2B) of a mouse wound after applying a gel containing dead bacteria culture according to an embodiment of the present invention.

[Fig. 3A] and [Fig. 3B] are bar graphs showing the staining results of wound tissue sections (Fig. 3A) and changes in wound distance of a mouse wound after applying a gel containing dead bacteria culture according to an embodiment of the present invention (Figure 3B).

[Fig. 4A] and [Fig. 4B] show the results of immunotissue staining of a mouse wound after applying a gel containing dead bacteria culture according to an embodiment of the present invention.

[Figure 5] shows the results of Western blot analysis showing that a dead Lactobacillus paracasei culture according to an embodiment of the present invention inhibits TGF-β-induced α-SMA expression pathway in vitro.

The singular forms "a", "an" and "the" mentioned in the present invention include plural references unless the context clearly dictates otherwise. The numerical range (such as 10%~11% of A) includes the upper and lower limits (ie 10% ≦A≦11%) unless otherwise specified; if the numerical range does not define the lower limit (such as less than 0.2% of B , Or B below 0.2%), it means that its lower limit may be 0 (ie 0%≦B≦0.2%). The above terms are used to describe and understand the present invention, but not to limit the present invention.

The present invention provides a local composition for promoting wound healing and reducing scars, which contains dead Lactobacillus culture as an effective ingredient, which can significantly promote wound healing and reduce scars.

In one embodiment, the dead Lactobacillus culture is derived from Lactobacillus paracasei GMNL-653. Specifically, the aforementioned Lactobacillus paracasei GMNL-653 was deposited at the Bioresource Conservation and Research Center (BCRC) of the Food Industry Development Institute, No. 331 Food Road, Hsinchu, Taiwan on February 26, 2016, and the deposit number is BCRC The strain of 910721.

In the above embodiment, the above dead bacteria culture of Lactobacillus comprises a pure dead bacteria culture solution and/or dry bacteria powder. In some examples, the above-mentioned pure culture solution of dead bacteria can be prepared, for example, by a conventional aseptic processing method, wherein the aseptic processing method can be, for example, chemical sterilization (such as ozone sterilization, ethylene oxide gas sterilization) or physical sterilization ( For example, ultraviolet irradiation, radiation irradiation, high temperature and high pressure sterilization, etc.). In other examples, the above-mentioned dried bacteria powder can be prepared by a conventional dehydration process, for example, the pure culture solution is processed by high temperature drying method, or the aseptically processed pure culture solution of dead bacteria is subjected to freeze vacuum drying method or spray drying method Manufactured by processing.

In application, the above-mentioned dead Lactobacillus culture can be used to manufacture local compositions and medical compositions that promote wound healing and reduce scars. In an embodiment applied to a topical composition, the topical composition may be, for example, a skin external composition. In an embodiment applied to a medical composition, the above-mentioned medical composition may include a dead lactobacillus culture with an effective dose of 1×10 ⁹ bacterial cells/mL to 1×10 ¹⁰ bacterial cells/mL. In one example, the above-mentioned pharmaceutical composition can be locally applied to the test site by means such as smearing, coating, etc. Therefore, the applicable dosage form can be, for example, a gel, a gel-like dressing, a sponge-like dressing, a film-like dressing, or any of the above. combination.

In the above embodiments, the carrier components used in the above dosage forms are not particularly limited, as long as they do not interfere with the efficacy of the dead Lactobacillus culture. In some examples, specific examples of the aforementioned carrier may include, but are not limited to, water, polymers, humectants, and the like. Specific examples of the aforementioned polymer may include, but are not limited to, xanthan gum. Specific examples of the aforementioned moisturizer may include, but are not limited to, xylitol, trehalose, glycerol and the like.

The above-mentioned topical composition containing dead Lactobacillus culture has been confirmed by animal experiments to promote wound healing and reduce scars. In one example, the aforementioned effect of wound healing may include, but is not limited to, promoting the expression level of type 1 matrix metalloproteinase-1 (MMP-1) in the early stage of wound healing, and increasing the level of wound healing in the middle and late stages of wound healing. The expression amount of type 1 collagen to accelerate wound healing.

In other examples, the aforementioned scar reduction effect may include, but is not limited to, inhibiting the overexpression of phosphorylation of Smad 2 and α-SMA induced by TGF-β, thereby regulating or avoiding scarring.

The above-mentioned topical composition containing dead bacteria culture of Lactobacillus of the present invention is a dead bacteria dosage form, which is relatively free from the problems of bacterial count and deterioration during application, and has better control in stability, and can be applied to manufacture and promote wounds Use of medical composition for healing and reducing scars.

Several embodiments are used below to illustrate the application of the present invention, but they are not used to limit the present invention. Those with ordinary knowledge in the technical field of the present invention can make various changes and modifications without departing from the spirit and scope of the present invention. Retouch.

Example 1. Screening of lactic acid bacteria that promote the secretion of collagen and MMP-1 expression in human skin fiber cells 1. Preparation of dead lactic acid bacteria culture

In this embodiment, Lactobacillus acidophilus , Lactobacillus casei , Lactobacillus fermentum , Lactobacillus paracasei , Lactobacillus plantarum , and rhamnosus are selected. Lactobacillus rhamnosus and many other strains were purchased from the Bioresource Conservation and Research Center (BCRC) of the Food Industry Development Institute, a consortium. Several strains were selected for each, and a total of 33 strains and target strains were tested. After culturing the cells of the tested strains overnight, wash them twice with sterile water, adjust the cell density to 1×10 ¹⁰ cells/mL, and then treat them at high temperature (121℃) and autoclave for 15 minutes to make them dead. Bacteria liquid is reserved.

The aforementioned target strain is Lactobacillus paracasei ( Lactobacillus paracasei ) GMNL-653, which is deposited at the Biological Resources Conservation and Research Center of the Food Industry Development Institute of the Republic of China. The deposit date is February 26, 2016, and the deposit number is BCRC 910721. Lactobacillus paracasei GMNL-653 is also deposited at the China Type Culture Collection (CCTCC), address: Wuhan, China, Wuhan University, zip code 430072, the deposit date is April 25, 2016, and the deposit number is CCTCC M2016226.

2. Establish cell test mode

The human skin HS68 fibroblast cell line (accession number: BCRC 60038; ATCC accession number: CRL-1635; CCTCC accession number: GDC403) was inoculated into a 6-well cell dish (2×10 ⁵ cells/well). After culturing overnight, wash twice with PBS, change to a serum-free cell culture medium and continue culturing. Cells of the experimental group are added with dead bacteria liquid of different strains (concentration of 1×10 ⁹ cells/mL, equivalent to 2× 10 ⁹ bacterial cells/well), while the control group cells were treated with sterile water equal in volume to the bacterial solution for 24 hours. After that, the supernatant is collected, and the supernatant is diluted 10-fold with dilution buffer, and then the commercially available enzyme immunoassay (EIA) kit for type 1 collagen pro-C-peptide (e.g., Procollagen type I C-peptide EIA kit; TaKaRa, #MK101) was analyzed, and the results are shown in Figure 1A.

Please refer to FIG. 1A, which is a bar graph of the relative expression level of type I collagen after co-cultivation of human skin HS68 fibroblast cell strain and different types of dead lactic acid bacteria strains according to an embodiment of the present invention . In this example, 34 strains were used for the in vitro model of human skin fibroblasts. Among them, 13 strains have the ability to promote collagen secretion. They are 1 strain of Lactobacillus acidophilus, 1 strain of Lactobacillus casei, 5 strains of Lactobacillus fermentum, and 3 strains. Strains of Lactobacillus paracasei (including the target strain GMNL-653 and 2 other strains), 2 strains of Lactobacillus plantarum, and 1 strain of Lactobacillus rhamnosus are shown in Figure 1A.

The results in Figure 1A show that the ability of Lactobacillus paracasei strain GMNL-653 to stimulate intracellular collagen secretion is better than that of the other 12 Lactobacillus strains. Therefore, the strain GMNL-653 was subsequently used for evaluation.

3. Steps to analyze the MMP-1 (Matrix metalloproteinase-1) gene content in human skin fiber cells

In this example, HS68 cell line was used for co-cultivation with different concentrations of Lactobacillus paracasei strain GMNL-653. First, wash the cells of Lactobacillus paracasei GMNL-653 cultured overnight with sterile water, adjust the cell density to 1×10 ¹⁰ cells/mL, and then sterilize them at high temperature (121°C) and autoclave for 15 Minutes later, make a dead bacteria culture for use.

The cell culture method of the HS68 cell line is the same as the second point of Example 1. The difference is that the HS68 cell line was treated with the dead bacteria liquid of strain GMNL-653 for 24 hours, the cell extract was collected, and the total RNA was extracted, and the reverse transcription reaction was performed to synthesize cDNA. The methods for extracting total RNA and reverse transcription reaction are well-known to those with ordinary knowledge in the technical field to which the present invention belongs, and will not be repeated here.

Then, quantitative PCR analysis of matrix metalloproteinase-1 (MMP-1) gene was performed using the primer pair shown in SEQ ID NO: 1 and 2, and β-actin (β-actin) The expression level of (using the primer pairs shown in SEQ ID NO: 3 and 4) was used as the endogenous control group, and the results are shown in Figure 1B.

Please refer to Figure 1B, which is a straight bar of the relative expression level of type 1 matrix metalloproteinase (MMP-1) after co-cultivation of a human skin HS68 fibroblast cell line with different concentrations of dead lactic acid bacteria strains according to an embodiment of the present invention In the figure, the figure number * represents the statistically significant difference between the experimental group and the control group after the student's t-test statistical analysis ( P <0.05).

The results of Fig. 1B show that the dead culture of Lactobacillus paracasei strain GMNL-653 can promote the expression of MMP-1 gene, and there is a statistically significant difference.

Example 2: Evaluation of the effect of Lactobacillus paracasei strain GMNL-653 in promoting wound healing 1. Preparation of gel containing dead bacteria culture

The Lactobacillus paracasei strain GMNL-653 was cultured overnight, washed twice with sterile water, adjusted to 2×10 ¹⁰ cells/mL, and sterilized at high temperature (121°C) and autoclaved for 15 minutes. Bacterial culture for use. Next, the dead bacteria culture of Lactobacillus paracasei strain GMNL-653 was transferred into the gel (containing 2% xanthan gum, 6% xylitol, 4% trehalose, 10% glycerol, and the rest was sterile Ultrapure water), and adjust the dead bacteria content to 1×10 ¹⁰ cells/g gel. The control group gel is the same formula gel without dead bacteria culture.

2. Establish an animal test model

Eight-week-old BALB/c mice (purchased from Lesco Biotech) were subjected to anesthesia and reflex tests. After confirming that they reached deep anesthesia, the following experiments were started. First, make a wound of about 0.2*1cm at a distance of about 1cm from the root of the tail of the mouse, and apply 0.01g of gel containing dead bacteria culture (the content of dead bacteria is 1×10 ¹⁰ cells/g of gel) or The control group gel, so the dosage of each mouse wound was equivalent to about 1×10 ⁸ bacterial cells. After application, let it stand for 15-20 minutes (at this time the mouse is in anesthetized state), so that the gel is completely absorbed. After applying it for 5 days, observe the wound recovery every 2-7 days, take pictures and use Image J imaging software to calculate the wound area for statistical analysis. The results are shown in Figure 2A and Figure 2B.

Please refer to FIGS. 2A and 2B, which show the wound appearance (FIG. 2A) and the wound size curve diagram (FIG. 2B) of a mouse wound after applying a gel containing dead bacteria culture according to an embodiment of the present invention. Figure 2B uses the wound size on the first day as 100% to evaluate the wound size of each group. The figure number * represents the statistical analysis of the student's t-test. Compared with the control group, the experimental group has a statistically significant difference ( P <0.05) (n=4).

The results of Fig. 2A and Fig. 2B show that the tail wounds of mice coated with Lactobacillus paracasei GMNL-653 gel healed faster than the control group. The difference can be seen from the 3rd day, and the observation point is reached on the 9th day It was more obvious on the 20th.

In addition, comparing the tail wounds on the 20th day, compared with the wounds of the control group, which left obvious scars after healing, the tail wounds of the mice coated with Lactobacillus paracasei GMNL-653 gel were smoother and had no obvious scars. , As shown in Figure 2A, shows that the dead culture of Lactobacillus paracasei GMNL-653 can indeed reduce scars.

3. Tissue staining analysis

The tissue samples from the wound area of the mouse tail at 5 and 9 days were collected, fixed, embedded, and sliced, and then stained with hematoxylin & eosin (HE). The results are shown in Figure 3A. The methods of fixing, embedding, slicing and HE staining are well known to those with ordinary knowledge in the technical field to which the present invention belongs, and will not be repeated here.

In addition, Image J software was used to measure the changes in the wound distance of the tail of the mouse, where the wound distance was defined as the distance between the intact hair follicles closest to both sides of the wound. The result is shown in Figure 3B.

Please refer to Figures 3A and 3B, which show the staining results of wound tissue sections (Figure 3A) and wound distance (unit: pixel) of a mouse wound after applying a gel containing dead bacteria cultures according to an embodiment of the present invention Histogram of changes (Figure 3B). The figure number * in Figure 3B represents the statistically significant difference between the experimental group and the control group after student's t-test statistical analysis ( P <0.05).

The results of Fig. 3A and Fig. 3B show that the mouse tail wounds coated with gel containing dead bacteria culture have significantly reduced the wound distance no matter from day 5 or to day 9, indicating that Lactobacillus paracasei GMNL- 653 dead bacteria culture can indeed accelerate wound healing.

Example 3: Evaluation of the mechanism of Lactobacillus paracasei strain GMNL-653 in promoting wound healing 1. Immunohistochemical staining (I)

In this example, tissue specimens from the wound area of the mouse tail on the 5th and 9th days of Example 2 were collected, fixed, embedded, and sectioned, and then subjected to immunotissue staining of wound healing related factors. Wound healing related factors include MMP-1, collagen (collagen) Masson's trichrome tissue staining, type 1 collagen (COL1A1), α-SMA and HE staining. The results are shown in Figure 4A and Figure 4B Shown. Relevant sections and all histological staining were commissioned by Bio-Life Technology Co., Ltd. (Taichung, Taiwan), and the experimental data were determined by professionals. The data of antibodies used to detect MMP-1, COL1A1 and α-SMA are shown in Table 1.

Please refer to FIG. 4A and FIG. 4B, which show the results of immunotissue staining of a mouse wound after smearing a gel containing dead bacteria culture according to an embodiment of the present invention.

The results of Fig. 4A and Fig. 4B show that the tail wounds of mice coated with gel containing dead bacteria culture can be observed at an earlier stage (the 5th day) in the tissue of matrix metalloproteinase-1 (MMP-1). There are many control groups, which means that the dead culture of Lactobacillus paracasei GMNL-653 can promote the expression of MMP-1 and accelerate the progress of the whole wound healing. In the later stage of wound healing (day 9), the dead bacteria culture of Lactobacillus paracasei GMNL-653 can promote the expression of collagen fibers and type 1 collagen, and accelerate the completion of wound healing.

α-SMA is an indicator of myofibroblast activation. Past studies have pointed out that the conversion of fibroblasts and myofibroblasts reaches a balance, which can prevent excessive scar formation. However, excessive activation of myofibroblasts can cause scarring to occur in large numbers. The results in Fig. 4B show that in the later stage of wound healing (the 9th day), the experimental group coated with a gel containing Lactobacillus paracasei GMNL-653 dead bacteria culture has less expression of α-SMA in the wound, which means that the paracase Lactobacillus GMNL-653 dead bacteria culture can effectively prevent the excessive activation of myofibroblasts, thereby avoiding the production of large amounts of scars.

2. Immunohistochemical staining (II)

This example uses the cell model to analyze the mechanism of the dead bacteria culture of Lactobacillus paracasei strain GMNL-653 in promoting wound healing. TGF-β is an important factor for activating myofibroblasts. However, in the wound remodeling phase, excessive TGF-β will cause excessive activation of myofibroblasts, leading to scarring.

Lactobacillus paracasei strain GMNL-653 was cultured overnight, washed twice with sterile water, adjusted the cell density to 1×10 ¹⁰ cells/mL, and then sterilized at high temperature (121℃) and autoclaved for 15 minutes. The dead bacteria culture is reserved. The cell culture method of HS68 cell line is the same as the second point of Example 1. The difference is that when changing to a serum-free cell culture medium to continue culturing, the cells of the experimental group are added with TGF-β (20ng/mL) and dead bacteria liquid (1×10 ⁹ cells/well or 2×10 ⁹ bacteria Somatic cells/well) for 24 hours. After that, the cell extract was collected, the protein was extracted and analyzed by Western blotting method for α-SMA, prototype Smad 2/3, phosphorylated-Smad2 (phospho-Smad 2; p-Smad 2) and housekeeping genes ( GAPDH; as the endogenous control group) protein content, the results are shown in Figure 5.

The extraction of proteins and western ink spotting methods are well known to those with ordinary knowledge in the technical field of the present invention, and will not be repeated here. The data of antibodies used to detect α-SMA, prototype Smad 2/3, p-Smad 2/3 and GAPDH are shown in Table 2.

Please refer to FIG. 5, which shows the results of Western blot analysis of the inhibition of TGF-β-induced α-SMA expression in vitro by a dead Lactobacillus paracasei culture according to an embodiment of the present invention.

The results in Fig. 5 show that the dead culture of Lactobacillus paracasei GMNL-653 can mainly inhibit the activated Smad 2 (ie p-Smad 2) induced by TGF-β, thereby affecting the expression of its downstream α-SMA. In other words, the dead culture of Lactobacillus paracasei GMNL-653 can inhibit the formation of scars by inhibiting the expression pathway of α-SMA induced by TGF-β.

It is supplemented that the present invention uses the dead bacteria culture of Lactobacillus paracasei GMNL-653 as the active ingredient, which can indeed accelerate wound healing, reduce the chance of pathogenic bacteria infecting the human body, and reduce scars. The mechanism may be promoted in the early stage of wound healing. The production of MMP-1 promotes the secretion of collagen in the middle and late stages of wound healing to accelerate wound healing. Furthermore, in vitro experiments have also confirmed that the dead culture of Lactobacillus paracasei GMNL-653 can inhibit the overexpression of p-Smad 2 and α-SMA induced by TGF-β, thereby reducing the production of scars. In addition, Lactobacillus paracasei GMNL-653 is a dead bacteria formulation, which is derived from the problem of sterile bacteria count and deterioration when added to subsequent related products. It can be better controlled in terms of stability and has a wide range of applications. It has the potential to be used in the manufacture of medical compositions that promote wound healing and reduce scars.

In summary, although the present invention takes a specific strain, a specific dosage form, a specific object, a specific administration method, or a specific evaluation method as an example, it illustrates the local composition and the pharmaceutical composition of the dead lactobacillus culture of the present invention. And its use in promoting wound healing and reducing scars, but anyone with ordinary knowledge in the technical field of the present invention knows that the present invention is not limited to this, and the present invention can also be used without departing from the spirit and scope of the present invention Other strains, other dosage forms, other objects, other administration methods or other evaluation methods.

It can be seen from the above examples that the advantages of the topical composition and the pharmaceutical composition of the present invention are that the active ingredient Lactobacillus paracasei GMNL-653 dead bacteria culture has excellent stability, safety, and does not have an abiotic count and The doubt of deterioration can significantly promote the ability of wound healing and reduce scars, and can be used in the manufacture of medical compositions that promote wound healing and reduce scars.

Although the present invention has been disclosed in several embodiments as above, it is not intended to limit the present invention. Anyone with ordinary knowledge in the technical field to which the present invention pertains can make various modifications without departing from the spirit and scope of the present invention. Modifications and modifications, therefore, the scope of protection of the present invention shall be subject to the scope of the attached patent application.

【Biological Material Deposit】

Lactobacillus paracasei GMNL-653 was deposited at the Biological Resources Conservation and Research Center of the Food Industry Development Institute of the Republic of China. The deposit date is February 26, 2016, and the deposit number is BCRC 910721.

<110> Jingyue Biotechnology Co., Ltd.

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<130> None

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<223> Downstream primer of MMP-1 gene

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Claims (3)

The use of a lactobacillus strain for the manufacture of a medicinal composition for promoting wound healing and reducing scars, wherein the medicinal composition contains a thermally lethal Lactobacillus body culture of Lactobacillus paracasei ( L. paracasei ) GMNL-653 as an effective Ingredients, the Lactobacillus paracasei GMNL-653 was deposited at the Bioresource Conservation and Research Center (BCRC), Food Industry Development Research Institute, No. 331 Food Road, Hsinchu, Taiwan on February 26, 2016, and the deposit number is BCRC 910721. The heat lethal Lactobacillus body culture is obtained after the Lactobacillus paracasei GMNL-653 is sterilized at 121°C for 15 minutes. The use of the lactobacillus strain described in item 1 of the scope of patent application for the manufacture of a pharmaceutical composition for promoting wound healing and reducing scars, wherein one of the effective doses of the heat-killed lactobacillus body culture is 1×10 ⁹ bacterial cells /mL to 1×10 ¹⁰ bacterial cells/mL. The use of the lactobacillus strain described in item 1 of the scope of patent application for the manufacture of a medical composition that promotes wound healing and reduces scars, wherein one of the dosage forms of the medical composition is a gel, a gel-like dressing, and a sponge Dressing, a film dressing or any combination of the above.

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