A skin cell dispersion for autologous treatment of skin wounds

Abstract

This invention relates to a skin cell dispersion for autologous treatment of skin wounds. Specifically, this invention provides a skin cell dispersion containing skin cells, epidermal growth factor, and transforming growth factor-β1. The skin cell dispersion of this invention exhibits excellent therapeutic effects on skin wounds.

Description

Technical Field

[0001] This invention belongs to the field of biotechnology, specifically relating to a skin cell dispersion for autologous treatment of skin wounds. Background Technology

[0002] For patients with skin burns, especially those with large-area burns, it is often necessary to take skin from other normal skin sites (i.e., donor sites) and then apply the harvested normal skin to the burned skin site to treat the burn.

[0003] The skin serves as a barrier to maintain internal environmental stability and prevent the invasion of microorganisms and chemicals. For burn patients, harvesting skin from other healthy tissues as donor sites results in donor wounds, especially for patients with large-area burns. These donor wounds disrupt the integrity of the skin, making them prone to infection and scarring. If the exposed wound is too large, bacteria can easily invade and multiply, causing electrolyte imbalances, anemia, high fever, and sepsis. Therefore, repairing donor wounds is crucial. However, current clinical practice lacks effective drugs for treating donor wounds, and most treatments rely on allowing the wound to heal naturally. However, this natural healing process has many drawbacks, such as slow healing rates, intense and prolonged pain for the patient, and a high risk of scarring. Furthermore, the slow healing rate often leads to microbial infections, causing further suffering for the patient.

[0004] Therefore, there is a need in this field to develop an effective drug for treating skin wounds. Summary of the Invention

[0005] The purpose of this invention is to provide a skin cell dispersion that can effectively treat skin wounds.

[0006] A first aspect of the present invention provides a skin cell dispersion containing skin cells, epidermal growth factor, and transforming growth factor-β1.

[0007] Preferably, the skin cell dispersion includes a skin cell dispersion for treating skin wounds.

[0008] Preferably, the epidermal growth factor includes human epidermal growth factor.

[0009] Preferably, the epidermal growth factor includes recombinant human epidermal growth factor.

[0010] Preferably, the transforming growth factor-β1 includes human transforming growth factor-β1.

[0011] Preferably, the transforming growth factor-β1 includes recombinant human transforming growth factor-β1.

[0012] Preferably, in the skin cell dispersion, the mass-volume concentration of the epidermal growth factor is 20-40 ng / ml, more preferably 25-35 ng / ml, more preferably 28-32 ng / ml, and more preferably 30 ng / ml.

[0013] Preferably, in the skin cell dispersion, the mass-volume concentration of transforming growth factor-β1 is 10-30 ng / ml, more preferably 15-25 ng / ml, more preferably 18-22 ng / ml, and more preferably 20 ng / ml.

[0014] Preferably, the weight ratio of the epidermal growth factor to the transforming growth factor-β1 is 0.5-5:1, more preferably 0.5-5:1, more preferably 1-3:1, more preferably 1-2:1, more preferably 1.3-1.7:1, and more preferably 1.5:1.

[0015] Preferably, the skin cell dispersion further includes a dispersion medium.

[0016] Preferably, the dispersion medium includes a cell dispersion medium.

[0017] Preferably, the dispersion medium comprises physiological saline or PBS buffered aqueous solution.

[0018] Preferably, the pH of the PBS buffer solution is 7.2-7.6, and more preferably 7.4.

[0019] Preferably, the skin cells include isolated or ex vivo skin cells.

[0020] Preferably, the skin comprises scaly skin.

[0021] Preferably, the skin includes human skin.

[0022] Preferably, the skin cells include scabra cells.

[0023] Preferably, the skin cells comprise cells obtained from scalpel skin dissection.

[0024] Preferably, the blade thickness skin includes normal blade thickness skin.

[0025] Preferably, the skin cells comprise normal sclerodermal cells.

[0026] Preferably, the blade-thick skin includes human blade-thick skin.

[0027] Preferably, the skin cells comprise skin cells obtained by dissociation from normal lateral thigh skin.

[0028] Preferably, the skin cells comprise normal skin cells from a patient with a skin wound.

[0029] Preferably, the skin cells include human skin cells or non-human mammalian (such as pig) skin cells.

[0030] Preferably, the skin cells include skin cells obtained after digesting the skin.

[0031] In a preferred example, the area (cm²) of the skin 2 The volume ratio (ml) of the skin cell dispersion to the volume ratio (cm) 2 The ratio ( / ml) is 1:0.5-1.5, preferably 1:0.8-1.2, and more preferably 1:1.

[0032] Preferably, the digestion is carried out in a digestive fluid.

[0033] Preferably, the skin is isolated or excised skin.

[0034] Preferably, the skin cells are prepared by the following method, which includes the steps of:

[0035] (1) After the skin is taken, it is digested with digestive juice to obtain digested skin pieces;

[0036] (2) Separate the cells from the digested skin to obtain skin cells.

[0037] Preferably, in step (1), the skin is separated or excised skin.

[0038] Preferably, in step (1), the skin is obtained using a dermabrasion knife.

[0039] Preferably, in step (1), the skin includes scaly skin.

[0040] Preferably, in step (1), the skin includes normal scaly skin.

[0041] Preferably, in step (1), the skin includes the skin of the normal outer thigh.

[0042] Preferably, in step (1), the skin includes human skin or non-human mammal (such as pig) skin.

[0043] Preferably, in step (1), the skin includes the normal skin of the patient with the skin wound.

[0044] Preferably, the digestive fluid includes trypsin and ethylenediaminetetraacetic acid.

[0045] Preferably, the trypsin has a weight percentage of 0.2-1.0 wt%, more preferably 0.2-0.8 wt%, more preferably 0.3-0.7 wt%, and more preferably 0.5 wt%, based on the weight of the digestive fluid.

[0046] Preferably, the ethylenediaminetetraacetic acid is 0.02-0.15 wt% by weight, more preferably 0.02-0.10 wt%, more preferably 0.04-0.06 wt%, more preferably 0.05 wt%, based on the weight of the digestive fluid.

[0047] Preferably, in step (1), the digestion temperature is 35-37°C, more preferably 37°C.

[0048] Preferably, in step (1), the digestion time is 15-25 min, more preferably 18-22 min, and more preferably 20 min.

[0049] Preferably, in step (1), the area (cm²) of the skin... 2 The ratio of the volume (ml) of the digestive fluid to the volume (cm) of the digestive fluid 2 The ratio of ( / ml) is 1:1-5, preferably 1:1.5-3.5, preferably 1:2-3, preferably 1:2.2-2.8, and preferably 1:2.5.

[0050] Preferably, in step (1), the skin is cleaned before digestion.

[0051] Preferably, in step (1), before digestion, the skin is washed with physiological saline or PBS 7.4 buffer.

[0052] Preferably, before step (2), the disassembled leather pieces are cleaned with a cleaning solution.

[0053] Preferably, the cleaning solution is used to wash away any remaining digestive fluid on the digested skin.

[0054] Preferably, the cleaning solution comprises a PBS buffered aqueous solution containing a soybean trypsin inhibitor.

[0055] Preferably, the pH of the PBS buffer solution is 7.2-7.6, and more preferably 7.4.

[0056] Preferably, the content of the soybean trypsin inhibitor is 0.05-0.15 wt%, more preferably 0.08-0.12 wt%, and more preferably 0.1 wt%, based on the weight of the cleaning solution.

[0057] Preferably, the cleaning temperature is 30-39℃, more preferably 35-37℃, and even more preferably 37℃.

[0058] Preferably, the area (cm²) of the skin in step (1) 2 The volume ratio (cm²) of the cleaning solution to the volume (ml) of the cleaning solution. 2 The ratio of ( / ml) is 1:1-5, preferably 1:1.5-3.5, preferably 1:2-3, preferably 1:2.2-2.8, and preferably 1:2.5.

[0059] Preferably, in step (2), the separation includes scraping.

[0060] Preferably, in step (2), the separation is performed by scraping with a scalpel.

[0061] Preferably, the skin cells obtained in step (2) are in the form of a cell suspension.

[0062] Preferably, the cell suspension includes a cell dispersion medium.

[0063] Preferably, the cell dispersion medium comprises physiological saline or PBS buffered aqueous solution.

[0064] Preferably, the pH of the PBS buffer solution is 7.2-7.6, and more preferably 7.4.

[0065] Preferably, step (2) includes the following steps:

[0066] After separating the cells from the digested skin flap, the separated cells are mixed with a cell dispersion medium, filtered, and a skin cell suspension is obtained. The obtained skin cells exist in the form of a cell suspension.

[0067] Preferably, the average viable cell yield in the cell suspension is 1.5%. 10 6 pcs / cm 2 -2.0 10 6 pcs / cm 2 The preferred value is 1.7. 10 6 pcs / cm 2 -2.0 10 6 pcs / cm 2 .

[0068] Preferably, the epidermal cell content in the skin cell dispersion is 1.5%. 10 6 pcs / cm 2 -2.0 10 6 / cm 2 The preferred value is 1.7. 10 6 pcs / cm 2 -2.0 10 6 pcs / cm 2 .

[0069] Preferably, the filter screen is a 90-110 mesh sieve, and more preferably a 100 mesh sieve.

[0070] In a preferred example, the area (cm²) of the skin in step (1) 2 The volume ratio (ml) of the skin cell dispersion to the volume ratio (cm) 2 The ratio ( / ml) is 1:0.5-1.5, preferably 1:0.8-1.2, and more preferably 1:1.

[0071] In a preferred example, the area (cm²) of the skin in step (1) 2 The mass ratio (ng) of the epidermal growth factor to the mass (cm) of the epidermal growth factor 2 The ratio of / ng) is 1:10-50, preferably 1:20-40, preferably 1:25-35, preferably 1:28-32, and preferably 1:30.

[0072] In a preferred example, the area (cm²) of the skin in step (1) 2 The mass ratio (ng) of the transforming growth factor-β1 to the mass ratio (cm) of the transforming growth factor-β1 2 The ratio of ( / ng) is 1:15-35, preferably 1:10-30, preferably 1:15-25, preferably 1:18-22, and preferably 1:20.

[0073] A second aspect of the present invention provides a method for preparing a skin cell dispersion as described in the first aspect of the present invention, the method comprising the steps of:

[0074] The skin cells were mixed with the epidermal growth factor and transforming growth factor-β1 to obtain the skin cell dispersion.

[0075] Preferably, the method includes the following steps:

[0076] The skin cells, epidermal growth factor, transforming growth factor-β1 and dispersion medium are mixed to obtain the skin cell dispersion.

[0077] Preferably, the method includes the following steps:

[0078] (1) After the skin is taken, it is digested with digestive juice to obtain digested skin pieces;

[0079] (2) Separate the cells from the digested skin flap to obtain skin cells;

[0080] (3) The skin cells, epidermal growth factor and transforming growth factor-β1 are mixed to obtain the cell dispersion.

[0081] Preferably, step (1) is as described in the first aspect of the present invention.

[0082] Preferably, step (2) is as described in the first aspect of the present invention.

[0083] Preferably, in step (3), the skin cells, epidermal growth factor, transforming growth factor-β1 and cell dispersion medium are mixed to obtain the skin cell dispersion.

[0084] Preferably, step (2) includes the following steps:

[0085] After separating the cells from the digested skin flap, the separated cells are mixed with a cell dispersion medium, filtered, and a skin cell suspension is obtained. The obtained skin cells exist in the form of a cell suspension.

[0086] Preferably, the filter screen is a 90-110 mesh sieve, and more preferably a 100 mesh sieve.

[0087] Preferably, the cell dispersion medium comprises physiological saline or PBS buffered aqueous solution.

[0088] Preferably, the pH of the PBS buffer solution is 7.2-7.6, and more preferably 7.4.

[0089] Preferably, step (3) includes:

[0090] (3) Skin cells in the form of cell suspension are mixed with epidermal growth factor and transforming growth factor-β1 to obtain the skin cell dispersion.

[0091] Preferably, the method includes the following steps:

[0092] (1) Take a thick piece of leather and place it in a digestive solution to digest it, thus obtaining a digested piece of leather;

[0093] (2) Separate the cells from the digested skin slices, mix the separated cells with a cell dispersion medium, filter, and obtain a skin cell suspension. The obtained skin cells exist in the form of a cell suspension.

[0094] (3) The skin cell suspension is mixed with the epidermal growth factor and transforming growth factor-β1 to obtain the skin cell dispersion.

[0095] Preferably, the method includes the following steps:

[0096] (1) Take (1.8-2.2) cm (1.8-2.2) cm thick leather pieces are placed in a digestive solution to digest them, resulting in digested leather pieces.

[0097] (2) After separating the cells from the digested skin slices, the separated cells are mixed with 3.5-4.5 ml of cell dispersion medium, filtered, and a skin cell suspension is obtained. The obtained skin cells exist in the form of a cell suspension.

[0098] (3) The skin cell suspension is mixed with the epidermal growth factor and transforming growth factor-β1 to obtain the skin cell dispersion, wherein the mass-volume concentration of epidermal growth factor in the skin cell dispersion is 28-32 ng / ml and the mass-volume concentration of transforming growth factor-β1 is 18-22 ng / ml.

[0099] Preferably, the method includes the following steps:

[0100] (1) Take (1.8-2.2) cm (1.8-2.2) cm thick leather pieces are placed in 9-11 ml of digestion solution and digested at 35-37℃ for 18-22 min to obtain digested leather pieces;

[0101] (2) After separating the cells from the digested skin slices, the separated cells are mixed with 3.5-4.5 ml of cell dispersion medium, filtered, and a skin cell suspension is obtained. The obtained skin cells exist in the form of a cell suspension.

[0102] (3) The skin cell suspension is mixed with the epidermal growth factor and transforming growth factor-β1 to obtain the skin cell dispersion, wherein the mass-volume concentration of epidermal growth factor in the skin cell dispersion is 28-32 ng / ml and the mass-volume concentration of transforming growth factor-β1 is 18-22 ng / ml.

[0103] A third aspect of the present invention provides a pharmaceutical composition comprising the skin cell dispersion as described in the first aspect of the present invention.

[0104] Preferably, the pharmaceutical composition further includes a pharmaceutically acceptable carrier.

[0105] Preferably, the dosage form of the pharmaceutical composition is a topical skin preparation.

[0106] Preferably, the dosage form of the pharmaceutical composition is a liquid, solid, or semi-solid formulation.

[0107] Preferably, the dosage form of the pharmaceutical composition is a spray, a film, an ointment, a cream, or a patch.

[0108] The present invention provides, in four aspects, the use of the skin cell dispersion as described in the first aspect of the present invention for preparing a composition for treating skin wounds.

[0109] Preferably, the skin comprises scaly skin.

[0110] Preferably, the skin wound includes a donor site wound.

[0111] Preferably, the skin wound includes a wound resulting from blade-thick skin removal.

[0112] Preferably, the skin wound includes a wound resulting from skin removal.

[0113] Preferably, the skin wound includes a wound resulting from normal blade-thickness skin removal.

[0114] Preferably, the skin wound includes a wound caused by the removal of normal skin.

[0115] Preferably, the skin removal includes blade-thickness skin removal.

[0116] Preferably, the skin wound includes a wound resulting from normal skin grafting in patients with deep second-degree burns.

[0117] Preferably, the skin wound includes a wound resulting from a normal blade-thickness skin graft in a patient with deep second-degree burns.

[0118] Preferably, the composition is a pharmaceutical composition.

[0119] Preferably, the composition further includes a pharmaceutically acceptable carrier.

[0120] Preferably, the dosage form of the composition is a topical skin preparation.

[0121] Preferably, the dosage form of the composition is a liquid, solid, or semi-solid formulation.

[0122] Preferably, the dosage form of the composition is a spray, a film-forming agent, an ointment, a cream, or a patch.

[0123] Preferably, the treatment includes autologous treatment.

[0124] Preferably, the composition is used to treat autologous skin wounds.

[0125] Preferably, the treatment of the skin wound includes promoting the healing of the skin wound and / or reducing the pain of the skin wound.

[0126] Preferably, the skin wound and the skin cells in the skin cell dispersion originate from the same object (or patient).

[0127] Preferably, the skin cells in the skin cell dispersion are derived from normal skin cells of the same skin wound object.

[0128] Preferably, the object includes a person.

[0129] The fifth aspect of the present invention provides a method for treating skin wounds by applying a skin cell dispersion as described in the first aspect of the present invention or a pharmaceutical composition as described in the third aspect of the present invention to a burn subject, thereby treating the skin wound.

[0130] Preferably, the application is topical.

[0131] Preferably, the application is by spraying or smearing.

[0132] It should be understood that the above-mentioned technical features of the present invention can be combined with each other to form new or preferred technical solutions. Attached Figure Description

[0133] Figure 1 Cell morphology of a cell suspension observed under an inverted phase-contrast microscope ( 100).

[0134] Figure 2 To observe the cell morphology of the cell suspension after 24 hours of adherence and growth under an inverted phase-contrast microscope ( 100).

[0135] Figure 3 The image shows a photograph of a skin burn before treatment of the donor site wound with the skin cell suspension prepared in Example 1.

[0136] Figure 4 Photographs showing the skin healing effect of the skin cell suspension prepared in Example 1 five days after autologous treatment of the donor site wound.

[0137] The present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention. Detailed Implementation

[0138] This invention discloses a skin cell dispersion containing skin cells, epidermal growth factor, and transforming growth factor-β1. The skin cell dispersion of this invention exhibits excellent therapeutic effects on skin wounds.

[0139] Experiments in this invention demonstrate that, for donor site wounds treated with autologous normal skin (such as split-thickness skin) transplantation from burn patients, the skin cell dispersion of this invention can significantly promote rapid wound healing and repair, shorten skin healing time, increase healing rate, reduce pain, etc., thereby achieving excellent wound treatment effects.

[0140] As used herein, the term "comprising" includes not only closed definitions but also semi-closed and open definitions. In other words, the term includes "consisting of" and "substantially consisting of".

[0141] As used in this article, the terms "ethylenediaminetetraacetic acid" and "EDTA" are used interchangeably.

[0142] As used in this article, the term “transforming growth factor-β1” is abbreviated as TGF-β1.

[0143] As used in this article, the term “recombinant human transforming growth factor-β1” is abbreviated as rhTGF-β1.

[0144] As used in this article, the term “epidermal growth factor” is abbreviated as EGF.

[0145] As used in this article, the term "recombinant human epidermal growth factor" is abbreviated as rh-EGF.

[0146] As used in this article, the English term "soybean trypsin inhibitor" is Trypsin inhibitor, soyabea.

[0147] As used in this article, the English term for "razor-thin graft" is razor-thin graft.

[0148] As used in this article, the English term "PBS" stands for phosphate buffered saline.

[0149] This invention provides a skin cell dispersion for the effective treatment of skin wounds.

[0150] This invention provides a skin cell dispersion containing skin cells, epidermal growth factor, and transforming growth factor-β1.

[0151] The skin cell dispersion of the present invention includes a skin cell dispersion for treating skin wounds.

[0152] In a preferred embodiment of the present invention, the epidermal growth factor in the skin cell dispersion has a mass-volume concentration of 20-40 ng / ml, preferably 25-35 ng / ml, preferably 28-32 ng / ml, and more preferably 30 ng / ml.

[0153] In a preferred embodiment of the present invention, the transforming growth factor-β1 concentration in the skin cell dispersion is 10-30 ng / ml, preferably 15-25 ng / ml, preferably 18-22 ng / ml, and preferably 20 ng / ml.

[0154] In a preferred embodiment of the present invention, the weight ratio of the epidermal growth factor to the transforming growth factor-β1 is 0.5-5:1, preferably 0.5-5:1, preferably 1-3:1, preferably 1-2:1, preferably 1.3-1.7:1, and preferably 1.5:1.

[0155] In a preferred embodiment of the present invention, the skin cell dispersion further includes a dispersion medium.

[0156] Preferably, the dispersion medium comprises physiological saline or PBS buffered aqueous solution.

[0157] In a preferred embodiment of the invention, the skin comprises scaly skin.

[0158] In a preferred embodiment of the invention, the skin cells include scabra cells.

[0159] In a preferred embodiment of the present invention, the skin cells include skin cells obtained after digesting the skin.

[0160] In a preferred example, the area (cm²) of the skin 2 The volume ratio (ml) of the skin cell dispersion to the volume ratio (cm) 2 The ratio ( / ml) is 1:0.5-1.5, preferably 1:0.8-1.2, and more preferably 1:1.

[0161] In a preferred embodiment of the present invention, the skin cells are prepared by the following method, the method comprising the steps of:

[0162] (1) After the skin is taken, it is digested with digestive juice to obtain digested skin pieces;

[0163] (2) Separate the cells from the digested skin flap to obtain skin cells.

[0164] Preferably, the digestive fluid includes trypsin and ethylenediaminetetraacetic acid.

[0165] Preferably, the trypsin has a weight percentage of 0.2-1.0 wt%, more preferably 0.2-0.8 wt%, more preferably 0.3-0.7 wt%, and more preferably 0.5 wt%, based on the weight of the digestive fluid.

[0166] Preferably, the ethylenediaminetetraacetic acid (EDTA) is 0.02-0.15 wt% by weight, more preferably 0.02-0.10 wt%, more preferably 0.04-0.06 wt%, and more preferably 0.05 wt% by weight of the digestive fluid.

[0167] Preferably, in step (1), the digestion temperature is 35-37°C, more preferably 37°C.

[0168] Preferably, in step (1), the digestion time is 15-25 min, more preferably 18-22 min, and more preferably 20 min.

[0169] Preferably, in step (2), the separation includes scraping.

[0170] Preferably, in step (2), the separation is performed by scraping with the blade of a scalpel.

[0171] Preferably, step (2) includes the following steps:

[0172] After separating the cells from the digested skin flap, the separated cells are mixed with a cell dispersion medium, filtered, and a skin cell suspension is obtained. The obtained skin cells exist in the form of a cell suspension.

[0173] Specifically, the skin cell dispersion is as described in the first aspect of the present invention above.

[0174] The present invention also provides a method for preparing the skin cell dispersion described herein, the method comprising the steps of:

[0175] The skin cells were mixed with the epidermal growth factor and transforming growth factor-β1 to obtain the skin cell dispersion.

[0176] In a preferred embodiment of the present invention, the method includes the steps of:

[0177] (1) After the skin is taken, it is digested with digestive juice to obtain digested skin pieces;

[0178] (2) Separate the cells from the digested skin to obtain skin cells.

[0179] (3) The skin cells, epidermal growth factor and transforming growth factor-β1 are mixed to obtain the cell dispersion.

[0180] Typically, the method includes the steps of:

[0181] (1) Take (1.8-2.2) cm (1.8-2.2) cm thick leather pieces are placed in a digestive solution to digest them, resulting in digested leather pieces.

[0182] (2) After separating the cells from the digested skin slices, the separated cells are mixed with 3.5-4.5 ml of cell dispersion medium, filtered, and a skin cell suspension is obtained. The obtained skin cells exist in the form of a cell suspension.

[0183] (3) The skin cell suspension is mixed with the epidermal growth factor and transforming growth factor-β1 to obtain the skin cell dispersion, wherein the mass-volume concentration of epidermal growth factor in the skin cell dispersion is 28-32 ng / ml and the mass-volume concentration of transforming growth factor-β1 is 18-22 ng / ml.

[0184] Specifically, the method for preparing the skin cell dispersion of the present invention is as described in the second aspect of the present invention above.

[0185] This invention provides a use of the aforementioned skin cell dispersion for preparing a composition for treating skin wounds.

[0186] In a preferred embodiment of the present invention, the skin wound includes a wound resulting from skin grafting.

[0187] In a preferred embodiment of the present invention, the composition is a pharmaceutical composition.

[0188] In a preferred embodiment of the present invention, the dosage form of the composition is a topical skin preparation.

[0189] Specifically, the use of the skin cell dispersion of the present invention is as described in the fourth aspect of the present invention above.

[0190] This invention provides a method for treating donor site wounds by applying a skin cell dispersion solution as described in this invention to a burn subject, thereby treating the skin wound.

[0191] Specifically, the method for treating donor site wounds according to the present invention is as described in the fifth aspect of the present invention above.

[0192] The present invention also provides a pharmaceutical composition comprising the skin cell dispersion described herein.

[0193] Preferably, the pharmaceutical composition further includes a pharmaceutically acceptable carrier.

[0194] Preferably, the dosage form of the pharmaceutical composition is a topical skin preparation.

[0195] Preferably, the dosage form of the pharmaceutical composition is a liquid, solid, or semi-solid formulation.

[0196] Preferably, the dosage form of the pharmaceutical composition is a spray, a film, an ointment, a cream, or a patch.

[0197] As used herein, "pharmaceutically acceptable carrier" refers to one or more compatible solid, semi-solid, liquid, or gel fillers that are suitable for human or animal use and must have sufficient purity and sufficiently low toxicity. "Compatibility" refers to the ability of the components in a pharmaceutical composition and the active ingredient of the drug, as well as their interactions, to not significantly reduce the efficacy of the drug.

[0198] It should be understood that, in this invention, there are no particular limitations on the pharmaceutically acceptable carriers used. Materials commonly used in the art can be selected, or they can be prepared using conventional methods or purchased from the market. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as methylcellulose, ethylcellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerin, mannitol, sorbitol, etc.), emulsifiers (such as Tween), wetting agents (such as sodium dodecyl sulfate), buffers, chelating agents, thickeners, pH adjusters, transdermal penetration enhancers, colorants, flavoring agents, stabilizers, antioxidants, preservatives, antibacterial agents, pyrogen-free water, etc.

[0199] In this invention, the dosage form of the composition includes, but is not limited to, a topical skin preparation. The topical skin preparations described in this invention are preferably preparations for use on burned skin.

[0200] Typically, the dosage form of the composition includes, but is not limited to, liquid, solid, or semi-solid formulations. For example, the dosage form of the composition may be a spray, film-forming agent, cream, lotion, or patch.

[0201] Pharmaceutical formulations should be matched to the route of administration, preferably for topical application, whereby a therapeutically effective amount of the drug is administered to the desired subject (e.g., human or non-human mammal). As used herein, the term "therapeutically effective amount" refers to an amount that is functionally or activityily produced in humans and / or animals and is acceptable to humans and / or animals. Those skilled in the art will understand that the "therapeutically effective amount" can vary depending on the form of the pharmaceutical composition, the route of administration, the excipients used, the severity of the disease, and whether it is used in combination with other drugs.

[0202] The safe and effective daily dosage of the active ingredient should take into account factors such as the route of administration and the health status of the patient / animal, all of which are within the skill range of a skilled physician / researcher.

[0203] The main advantages of this invention include:

[0204] 1. The skin cell dispersion of the present invention has excellent therapeutic effects on skin wounds. For burn patients who have undergone autologous normal skin (donor site) transplantation, the skin cell dispersion of the present invention can significantly promote rapid healing and repair of the donor site wound, shorten skin healing time, improve healing rate, and reduce pain, thus exhibiting excellent therapeutic effects on skin wounds.

[0205] 2. The skin cells in the skin cell dispersion of the present invention can be directly derived from cells dissociated and collected from the skin (such as scalp dermal filler). There is no need for cell culture in vitro. It can be directly sprayed on the donor site wound, which significantly promotes the rapid regeneration and healing of the donor site wound, thereby improving the treatment efficiency of the skin cell dispersion on the donor site wound, reducing operation time, and making the operation simple and convenient, reducing costs.

[0206] 3. The skin cell dispersion of the present invention can significantly increase the area of ​​the cell dispersion prepared by skin harvesting per unit area that can be applied to the donor site wound, enabling the treatment of a larger donor site wound with a smaller donor site, thereby reducing the amount of skin harvested by the patient, reducing patient suffering, and significantly improving treatment efficiency.

[0207] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention.

[0208] Example 1: Skin cell dispersion

[0209] 1. Preparation of skin cell dispersion:

[0210] (1) Trypsin and ethylenediaminetetraacetic acid were mixed in physiological saline to obtain cell digestion solution, wherein the weight percentage of trypsin was 0.50 wt% and the weight percentage of ethylenediaminetetraacetic acid was 0.05 wt%, based on the weight of the cell digestion solution.

[0211] (2) Using a dermatome, a 2cm thick, normal, non-wound skin graft is taken from the lateral thigh of a patient with deep second-degree burns. After washing with physiological saline, the cut-thickness skin piece was placed in 10 ml of the cell digestion solution prepared in step (1) and digested at 37°C for 20 min to obtain the digested skin piece.

[0212] (3) Use sterile forceps to remove the digested skin pieces and immerse them in 10 ml of PBS 7.4 buffer solution containing 0.1 wt% soybean trypsin inhibitor at 37°C to wash away the residual cell digestion solution and obtain the washed skin pieces.

[0213] (4) Place the cleaned skin patch in a sterile tray, fix the skin patch with medical tweezers, scrape off the cells with a scalpel, and then wash the scalpel and tray with 4ml PBS 7.4 buffer solution to obtain a cell suspension. Collect the cell suspension in one corner of the tray, collect the cell suspension with a syringe, and filter it through a 100-mesh sieve to obtain the filtered cell suspension.

[0214] (5) The filtered cell suspension obtained in step (4) is mixed with recombinant human epidermal growth factor (rh-EGF) and recombinant human transforming growth factor-β1 (rhTGF-β1) to obtain a skin cell dispersion, wherein the mass-volume concentration of recombinant human epidermal growth factor (rh-EGF) in the skin cell dispersion is 30 ng / ml and the mass-volume concentration of recombinant human transforming growth factor-β1 (rhTGF-β1) is 20 ng / ml.

[0215] Growth characteristics of cell suspension

[0216] The cell morphology of the filtered cell suspension obtained in step (4) is as follows: Figure 1 As shown, Figure 1 The cells showed good growth, with an average viable cell yield of 1.85%. 10 6 pcs / cm 2 The cell viability was 83%.

[0217] The cell morphology of the filtered cell suspension obtained in step (4) after 24 hours of adherent growth in serum-free culture medium is as follows: Figure 2 As shown, Figure 2 The cells exhibit good adherent growth and clonate aggregation, indicating excellent adherent growth characteristics and thus superior regenerative and repair capabilities in skin wounds.

[0218] Example 2: Skin cell suspension

[0219] Preparation of skin cell suspension:

[0220] (1) Trypsin and ethylenediaminetetraacetic acid were mixed in physiological saline to obtain cell digestion solution, wherein the weight percentage of trypsin was 0.50 wt% and the weight percentage of ethylenediaminetetraacetic acid was 0.05 wt%, based on the weight of the cell digestion solution.

[0221] (2) Using a dermatome, a 2cm thick, normal, non-wound skin graft is taken from the lateral thigh of a patient with deep second-degree burns. After washing with physiological saline, the cut-thickness skin piece was placed in 10 ml of the cell digestion solution prepared in step (1) and digested at 37°C for 20 min to obtain the digested skin piece.

[0222] (3) Use sterile forceps to remove the digested skin pieces and immerse them in 10 ml of PBS 7.4 buffer solution containing 0.1 wt% soybean trypsin inhibitor at 37°C to wash away the residual cell digestion solution and obtain the washed skin pieces.

[0223] (4) Place the cleaned skin patch in a sterile tray, fix the skin patch with medical tweezers, scrape off the cells with a scalpel, and then clean the scalpel and tray with 4ml PBS 7.4 buffer solution to obtain a cell suspension. Collect the cell suspension in one corner of the tray, collect the cell suspension with a syringe, filter it through a 100-mesh sieve, and obtain the filtered cell suspension, which is the skin cell suspension.

[0224] Example 3: Skin Cell Dispersion

[0225] Preparation of skin cell dispersion:

[0226] (1) Trypsin and ethylenediaminetetraacetic acid were mixed in physiological saline to obtain cell digestion solution, wherein the weight percentage of trypsin was 0.50 wt% and the weight percentage of ethylenediaminetetraacetic acid was 0.05 wt%, based on the weight of the cell digestion solution.

[0227] (2) Using a dermatome, a 2cm thick, normal, non-wound skin graft is taken from the lateral thigh of a patient with deep second-degree burns. After washing with physiological saline, the cut-thickness skin piece was placed in 10 ml of the cell digestion solution prepared in step (1) and digested at 37°C for 20 min to obtain the digested skin piece.

[0228] (3) Use sterile forceps to remove the digested skin pieces and immerse them in 10 ml of PBS 7.4 buffer solution containing 0.1 wt% soybean trypsin inhibitor at 37°C to wash away the residual cell digestion solution and obtain the washed skin pieces.

[0229] (4) Place the cleaned skin patch in a sterile tray, fix the skin patch with medical tweezers, scrape off the cells with a scalpel, and then wash the scalpel and tray with 4ml PBS 7.4 buffer solution to obtain a cell suspension. Collect the cell suspension in one corner of the tray, collect the cell suspension with a syringe, and filter it through a 100-mesh sieve to obtain the filtered cell suspension.

[0230] (5) The filtered cell suspension obtained in step (4) is mixed with recombinant human transforming growth factor-β1 (rhTGF-β1) to obtain a skin cell dispersion, wherein the mass-volume concentration of recombinant human transforming growth factor-β1 (rhTGF-β1) in the skin cell dispersion is 40 ng / ml.

[0231] Example 4: Skin Cell Dispersion

[0232] Preparation of skin cell dispersion:

[0233] (1) Trypsin and ethylenediaminetetraacetic acid were mixed in physiological saline to obtain cell digestion solution, wherein the weight percentage of trypsin was 0.50 wt% and the weight percentage of ethylenediaminetetraacetic acid was 0.05 wt%, based on the weight of the cell digestion solution.

[0234] (2) Using a dermatome, a 2cm thick, normal, non-wound skin graft is taken from the lateral thigh of a patient with deep second-degree burns. After washing with physiological saline, the cut-thickness skin piece was placed in 10 ml of the cell digestion solution prepared in step (1) and digested at 37°C for 20 min to obtain the digested skin piece.

[0235] (3) Use sterile forceps to remove the digested skin pieces and immerse them in 10 ml of PBS 7.4 buffer solution containing 0.1 wt% soybean trypsin inhibitor at 37°C to wash away the residual cell digestion solution and obtain the washed skin pieces.

[0236] (4) Place the cleaned skin patch in a sterile tray, fix the skin patch with medical tweezers, scrape off the cells with a scalpel, and then wash the scalpel and tray with 4ml PBS 7.4 buffer solution to obtain a cell suspension. Collect the cell suspension in one corner of the tray, collect the cell suspension with a syringe, and filter it through a 100-mesh sieve to obtain the filtered cell suspension.

[0237] (5) The filtered cell suspension obtained in step (4) is mixed with recombinant human epidermal growth factor (rh-EGF) to obtain a skin cell dispersion, wherein the mass-volume concentration of recombinant human epidermal growth factor (rh-EGF) in the skin cell dispersion is 60 ng / ml.

[0238] Example 5: Skin Cell Dispersion

[0239] Preparation of skin cell dispersion

[0240] (1) Trypsin and ethylenediaminetetraacetic acid were mixed in physiological saline to obtain cell digestion solution, wherein the weight percentage of trypsin was 0.50 wt% and the weight percentage of ethylenediaminetetraacetic acid was 0.05 wt%, based on the weight of the cell digestion solution.

[0241] (2) Using a dermatome, a 2cm thick, normal, non-wound skin graft is taken from the lateral thigh of a patient with deep second-degree burns. After washing with physiological saline, the cut-thickness skin piece was placed in 10 ml of the cell digestion solution prepared in step (1) and digested at 37°C for 20 min to obtain the digested skin piece.

[0242] (3) Use sterile forceps to remove the digested skin pieces and immerse them in 10 ml of PBS 7.4 buffer solution containing 0.1 wt% soybean trypsin inhibitor at 37°C to wash away the residual cell digestion solution and obtain the washed skin pieces.

[0243] (4) Place the cleaned skin patch in a sterile tray, fix the skin patch with medical tweezers, scrape off the cells with a scalpel, and then wash the scalpel and tray with 4ml PBS 7.4 buffer solution to obtain a cell suspension. Collect the cell suspension in one corner of the tray, collect the cell suspension with a syringe, and filter it through a 100-mesh sieve to obtain the filtered cell suspension.

[0244] (5) The filtered cell suspension obtained in step (4) is mixed with recombinant human vascular endothelial growth factor (rh-VEGF) and recombinant human transforming growth factor-β1 (rhTGF-β1) to obtain a skin cell dispersion, wherein the mass-volume concentration of recombinant human vascular endothelial growth factor (rh-VEGF) in the skin cell dispersion is 30 ng / ml and the mass-volume concentration of recombinant human transforming growth factor-β1 (rhTGF-β1) is 20 ng / ml.

[0245] Example 6: Skin Cell Dispersion

[0246] Preparation of skin cell dispersion

[0247] (1) Trypsin and ethylenediaminetetraacetic acid were mixed in physiological saline to obtain cell digestion solution, wherein the weight percentage of trypsin was 0.50 wt% and the weight percentage of ethylenediaminetetraacetic acid was 0.05 wt%, based on the weight of the cell digestion solution.

[0248] (2) Using a dermatome, a 2cm thick, normal, non-wound skin graft is taken from the lateral thigh of a patient with deep second-degree burns. After washing with physiological saline, the cut-thickness skin piece was placed in 10 ml of the cell digestion solution prepared in step (1) and digested at 37°C for 20 min to obtain the digested skin piece.

[0249] (3) Use sterile forceps to remove the digested skin pieces and immerse them in 10 ml of PBS 7.4 buffer solution containing 0.1 wt% soybean trypsin inhibitor at 37°C to wash away the residual cell digestion solution and obtain the washed skin pieces.

[0250] (4) Place the cleaned skin patch in a sterile tray, fix the skin patch with medical tweezers, scrape off the cells with a scalpel, and then wash the scalpel and tray with 4ml PBS 7.4 buffer solution to obtain a cell suspension. Collect the cell suspension in one corner of the tray, collect the cell suspension with a syringe, and filter it through a 100-mesh sieve to obtain the filtered cell suspension.

[0251] (5) The filtered cell suspension obtained in step (4) is mixed with recombinant human epidermal growth factor (rh-EGF) and recombinant human transforming growth factor-β1 (rhTGF-β1) to obtain a skin cell dispersion, wherein the mass-volume concentration of recombinant human epidermal growth factor (rh-EGF) in the skin cell dispersion is 8 ng / ml and the mass-volume concentration of recombinant human transforming growth factor-β1 (rhTGF-β1) is 40 ng / ml.

[0252] Example 7 Skin Cell Dispersion

[0253] Preparation of skin cell dispersion

[0254] (1) Trypsin and ethylenediaminetetraacetic acid were mixed in physiological saline to obtain cell digestion solution, wherein the weight percentage of trypsin was 0.50 wt% and the weight percentage of ethylenediaminetetraacetic acid was 0.05 wt%, based on the weight of the cell digestion solution.

[0255] (2) Take a 2cm full-thickness skin graft from the lateral thigh of a patient with deep second-degree burns using a dermatome. After washing with physiological saline, the cut-thickness skin piece was placed in 10 ml of the cell digestion solution prepared in step (1) and digested at 37°C for 20 min to obtain the digested skin piece.

[0256] (3) Use sterile forceps to remove the digested skin pieces and immerse them in 10 ml of PBS 7.4 buffer solution containing 0.1 wt% soybean trypsin inhibitor at 37°C to wash away the residual cell digestion solution and obtain the washed skin pieces.

[0257] (4) Place the cleaned skin patch in a sterile tray, fix the skin patch with medical tweezers, scrape off the cells with a scalpel, and then wash the scalpel and tray with 4ml PBS 7.4 buffer solution to obtain a cell suspension. Collect the cell suspension in one corner of the tray, collect the cell suspension with a syringe, and filter it through a 100-mesh sieve to obtain the filtered cell suspension.

[0258] (5) The filtered cell suspension obtained in step (4) is mixed with recombinant human epidermal growth factor (rh-EGF) and recombinant human transforming growth factor-β1 (rhTGF-β1) to obtain a skin cell dispersion, wherein the mass-volume concentration of recombinant human epidermal growth factor (rh-EGF) in the skin cell dispersion is 30 ng / ml and the mass-volume concentration of recombinant human transforming growth factor-β1 (rhTGF-β1) is 20 ng / ml.

[0259] Effect Example

[0260] The skin cell dispersions prepared in Examples 1 and 3-7 and the skin cell suspension prepared in Example 2 were examined for use in autologous treatment of donor site wounds.

[0261] 1. Method

[0262] Following burn treatment using autologous normal-thickness skin grafts (donor sites) transplanted to the burn wounds of deep second-degree burn patients, donor site wounds appeared at the site of autologous normal-thickness skin graft removal. Skin cell dispersions prepared from normal, un-wounded skin grafts from the lateral thigh of the same deep second-degree burn patient, as well as a skin cell suspension from Example 2, were sprayed onto the donor site wounds appearing at the autologous normal-thickness skin graft removal site. The spraying volume was based on a 1cm graft. Skin cell dispersions and suspensions prepared from 1cm thick skin grafts were evenly sprayed onto an 80cm surface. 2 The donor site wound was sprayed and then covered and bandaged with sterile dressing.

[0263] 2. Evaluation of treatment efficacy

[0264] The complete wound healing rate and pain index were used to investigate the autologous therapeutic effect of skin cell dispersions prepared in Examples 1 and 3-7 and skin cell suspensions prepared in Example 2 on the donor site wounds resulting from autologous transplantation of normal skin (donor site) to the burn wounds of patients with the same deep second-degree burns.

[0265] 2.1 Evaluation of wound healing rate

[0266] The wound healing rate of the skin cell dispersions prepared in Examples 1, 3-7 and the skin cell suspension prepared in Example 2 on day 5 after autologous treatment of donor site wounds was evaluated.

[0267] Wound healing evaluation criteria: The wound surface is epithelialized, there is no exudate, and no drainage or dressing is required.

[0268] Wound healing rate = (initial wound contour area - postoperative unhealed wound contour area) / initial wound contour area × 100%.

[0269] In each embodiment, before and 5 days after autologous treatment of the donor site wound, the skin cell dispersion and skin cell suspension were used. The wound was covered with sterile plastic film, and its outline was drawn. The drawn sterile plastic film was then scanned into a computer, and the wound length (cm), width (cm), and outline area (cm²) were calculated and analyzed using image analysis software (Adobe Photoshop 7.0 and ImageJ version 1.46j). 2 The study evaluated the effects of skin cell dispersions and skin cell suspensions of each embodiment on skin wound healing after autologous treatment of donor sites.

[0270] Table 1 shows the wound healing rates on day 5 after autologous treatment of donor site wounds with the skin cell dispersions prepared in Examples 1, 3-7, and the skin cell suspension prepared in Example 2.

[0271] Table 1. Wound healing rates on day 5 after autologous treatment of donor site wounds using skin cell dispersions prepared in Examples 1, 3-7 and skin cell suspension prepared in Example 2 (n=10).

[0272]

[0273] As shown in Table 1, the skin cell dispersion and skin cell suspension prepared in this embodiment can significantly promote the regeneration and healing capacity of the donor site wound. Epidermal growth factor and transforming growth factor-β1 can synergistically enhance the therapeutic effect of cells from split-thickness skin grafts on the donor site wound. The before-and-after photos of the effects of the skin cell dispersion prepared in Example 1 on autologous treatment of the donor site wound are shown in the table. Figure 3 and Figure 4 As shown, from Figure 3-4 As can be seen, the skin cell dispersion of Example 1 can significantly promote the healing of the donor site wound, thus having a better therapeutic effect on the donor site wound.

[0274] 2.2 Pain Assessment

[0275] The subjective pain score (NRS score) of patients was evaluated 5 days after autologous treatment of donor site wounds with skin cell dispersions prepared in Examples 1, 3-7 and skin cell suspensions prepared in Example 2.

[0276] NRS scoring criteria: The Numerical Rating Scale (NRS) uses a 10 cm long pain scale, where 0 represents no pain and 10 represents the most painful. Patients choose one number from these 11 numbers to represent their pain level.

[0277] NRS pain scoring system (0-10 points):

[0278]

[0279] 0 points: Painless;

[0280] 1-3 points: Mild pain, sleep unaffected;

[0281] 4-6 points: Moderate pain, sleep disturbance;

[0282] 7-9 points: Severe pain, seriously affecting sleep;

[0283] 10 / 10: Extreme pain.

[0284] The higher the score, the more severe the pain.

[0285] Table 2 shows the subjective pain scores (NRS scores) of patients 5 days after autologous treatment of donor site wounds with the skin cell dispersions prepared in Examples 1, 3-7, and the skin cell suspension prepared in Example 2.

[0286] Table 2. Subjective pain ratings (NRS scores) of patients 5 days after autologous treatment of donor wounds with skin cell dispersions prepared in Examples 1, 3-7 and skin cell suspension prepared in Example 2 (n=10).

[0287]

[0288] As can be seen from Table 2, the skin cell dispersion prepared in Example 1 can significantly reduce the pain caused by the donor site wound, thus indicating that the skin cell dispersion suspension prepared in Example 1 has excellent therapeutic effect on skin trauma.

[0289] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A skin cell dispersion liquid, characterized by, The skin cell dispersion liquid contains skin cells, epidermal growth factor and transforming growth factor-β1. The skin cell dispersion liquid is prepared by the following method: (1) taking a split-thickness skin graft, and digesting the split-thickness skin graft in a digestive solution to obtain a digested skin graft; (2) separating cells of the digested skin graft, and mixing the separated cells with a cell dispersion medium, and filtering to obtain a skin cell suspension; (3) mixing the skin cell suspension with the epidermal growth factor and the transforming growth factor-β1 to obtain the skin cell dispersion liquid, wherein the mass-volume concentration of the epidermal growth factor in the skin cell dispersion liquid is 28-32 ng / ml, and the mass-volume concentration of the transforming growth factor-β1 in the skin cell dispersion liquid is 18-22 ng / ml.

2. The skin cell dispersion of claim 1, wherein, In the skin cell dispersion liquid, the mass-volume concentration of the epidermal growth factor is 30 ng / ml. In the skin cell dispersion liquid, the mass-volume concentration of the transforming growth factor-β1 is 20 ng / ml.

3. The skin cell dispersion of claim 1, wherein the dispersion is in a medium comprising a serum-free cell culture medium. The weight ratio of the epidermal growth factor to the transforming growth factor-β1 is 1.5:

1.

4. The skin cell dispersion of claim 1, wherein the dispersion is in a medium comprising a serum-free cell culture medium. The cell dispersion medium comprises physiological saline or a PBS aqueous buffer solution.

5. The skin cell dispersion of claim 1, wherein the dispersion is in a medium comprising a serum-free cell culture medium. The area of the split-thickness skin graft to the volume of the skin cell dispersion is 1 cm 2 : (0.8 ml - 1.2 ml).

6. The skin cell dispersion of claim 1, wherein the dispersion is in a medium comprising a serum-free cell culture medium. In the step (1), the digestive solution comprises trypsin and ethylenediaminetetraacetic acid.

7. The skin cell dispersion of claim 6, wherein the dispersion is in a medium comprising a serum-free cell culture medium. The weight percentage of the trypsin is 0.3-0.7 wt% based on the weight of the digestive solution; and The weight percentage of the ethylenediaminetetraacetic acid is 0.04-0.06 wt% based on the weight of the digestive solution.

8. The skin cell dispersion of claim 1, wherein the dispersion is in a medium comprising a serum-free cell culture medium. In the step (1), the area of the split-thickness skin graft is 1 cm 2 : (2.2 ml-2.8 ml).

9. A pharmaceutical composition, characterized by, The pharmaceutical composition comprises the skin cell dispersion liquid according to claim 1.

10. The pharmaceutical composition of claim 9, wherein The pharmaceutical composition further comprises a pharmaceutically acceptable carrier.

11. The pharmaceutical composition of claim 9, wherein The pharmaceutical composition is a skin external preparation.

12. The pharmaceutical composition of claim 9, wherein The pharmaceutical composition is a spray, a film-coating agent, a cream, a paste or a patch.

13. Use of the skin cell dispersion liquid according to claim 1 in the preparation of a composition for treating a skin wound surface.

14. Use according to claim 13, characterized in that, The skin wound surface includes a wound surface caused by skin removal.

15. The use according to claim 13, characterized in that, The skin wound surface includes a wound surface caused by split-thickness skin graft removal.

16. The use according to claim 13, characterized in that, The composition is a pharmaceutical composition.

17. The use according to claim 13, characterized in that, The composition is a skin external preparation.

18. The use of claim 13, wherein, The composition is a spray, a film-coating agent, a cream, a paste or a patch.

19. The use of claim 13, wherein, The treatment of the skin wound surface includes promoting the healing of the skin wound surface and / or reducing the pain of the skin wound surface.

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