Medicine for accelerating skin repair and regeneration, preparation method thereof and application thereof

Abstract

The invention discloses a medicine for accelerating the repair of stratum epidermis and the regeneration of skin appendages. The medicine consists of epidermal stem cells, bone marrow mesenchymal stem cells and a medicine carrier, wherein the epidermal stem cells and the bone marrow mesenchymal stem cells are adhered to the medicine carrier, namely each cubic millimeter of the medicine carrier contains (0.3-1)*104 epidermal stem cells and (0.3-1)*104 bone marrow mesenchymal stem cells. The medicine has an obvious effect in aspects of promoting the regeneration of the stratum epidermis, promoting the regeneration of dermis, enhancing the mechanical strength of regenerated skin, promoting the content of collagen of the regenerated skin, promoting revascularization, promoting the regeneration of hair follicles and the like, so that wounds can be repaired within a short time and the regeneration of the skin appendages can be accelerated. The invention also discloses a method for preparing the medicine for accelerating the repair of the stratum epidermis and the regeneration of the skin appendages, and the method is simple, high in controllability, high in operability and high in repeatability.

Description

technical field

[0001] The invention relates to a medicine for accelerating skin layer repair and skin accessory organ regeneration, in particular to a medicine composed of epidermal stem cells, bone marrow mesenchymal stem cells and carriers. Background technique

[0002] The skin is the largest organ of the human body, covering the whole body, and it can protect various tissues and organs in the body from physical damage, mechanical damage, chemical damage and biological damage. Skin regeneration mainly includes: 1) regeneration of various skin cells; 2) regeneration and remodeling of skin extracellular matrix, including various collagen and other components; 3) regeneration and functional recovery of skin appendages. With the continuous in-depth understanding of the mechanism of skin regeneration, the promotion of skin regeneration has become an important guide for skin repair and beauty, breaking the traditional old concept of "scars will inevitably be produced after wou...

Examples

Embodiment 1

[0042] Example 1: Preparation method, optimization of preparation conditions and identification of epidermal stem cells

[0043] 1. Preparation method of epidermal stem cells

[0044] At present, the preparation method of epidermal stem cells mainly includes two steps: 1) Firstly, a single skin epidermis is obtained through the functional separation of the junction between the epidermis and the dermis by neutral protease, which avoids the pollution of dermal cells; and, because trypsin damages the cells Therefore, the neutral protease digestion method is firstly used to separate the epidermal layer of the skin by using its function of hydrolyzing the junction between the epidermis and the dermis, and then the epidermal layer of the skin is digested with trypsin to obtain a single cell. It is beneficial to the protection of the activity of the digested cells and ensures that a sufficient number of active target cells are obtained; 2) the epidermal stem cells are then separated ...

Embodiment 2

[0099] Example 2: Preparation of bone marrow mesenchymal stem cells

[0100] The preparation of bone marrow mesenchymal stem cells can refer to the literature (Cai-Xia He, Ni Li, Yu-Lan Hu, Xiu-Mei Zhu, Hai-Jie Li, Min Han, Pei-Hong Miao, Zhong-Jie Hu, Gang Wang, Wen-Quan Liang, Yasuhiko Tabata, Jian-Qing Gao*, Effective gene delivery to mesenchymal stem cells based on the novel reverse transfection and three-dimensional cell culture system, Pharm Res, 2011, 28(7): 1577-1590)

[0101] 1) Preparation and in vitro culture of primary rat bone marrow mesenchymal stem cells (BMSCs): 3-week-old SD rats were sacrificed by neck pulling, the tibia and femur were separated, and the containing The tibia and femur were repeatedly washed with serum culture solution, and the obtained bone marrow was passed through a 200-mesh cell sieve (Shanghai Ruigu Biotechnology Co., Ltd.), centrifuged at 1000 rpm for 5 minutes, and the supernatant was discarded. After centrifugation, add the DMEM low-s...

Embodiment 3

[0104] Embodiment 3: the preparation of the medicine that accelerates skin layer repair and skin accessory organ regeneration

[0105] 1) The epidermal stem cells (ESCs) prepared by experimental group 7# in Example 1 were first washed twice with PBS, and then washed with trypsin-ethylenediaminetetraacetic acid (Trypsin-EDTA) containing 0.05% by weight of pancreatic Enzyme cell digestion solution (trypsin cell digestion solution obtained by dissolving trypsin-ethylenediaminetetraacetic acid in phosphate buffer at pH = 7.4) was digested for 7 minutes, and then added to the epidermal stem cells digested by trypsin cell digestion solution containing The DMEM low-sugar / F12 culture fluid of fetal bovine serum and penicillin streptomycin (the DMEM low-sugar / F12 culture fluid that contains fetal bovine serum and penicillin streptomycin is made of 10% fetal bovine serum by weight percentage, and the weight percentage is 1% penicillin, streptomycin, DMEM low-sugar culture medium of 59.3...

technical field

[0001] The invention relates to a medicine for accelerating skin layer repair and skin accessory organ regeneration, in particular to a medicine composed of epidermal stem cells, bone marrow mesenchymal stem cells and carriers. Background technique

[0002] The skin is the largest organ of the human body, covering the whole body, and it can protect various tissues and organs in the body from physical damage, mechanical damage, chemical damage and biological damage. Skin regeneration mainly includes: 1) regeneration of various skin cells; 2) regeneration and remodeling of skin extracellular matrix, including various collagen and other components; 3) regeneration and functional recovery of skin appendages. With the continuous in-depth understanding of the mechanism of skin regeneration, the promotion of skin regeneration has become an important guide for skin repair and beauty, breaking the traditional old concept of "scars will inevitably be produced after wou...

Examples

Embodiment 1

[0042] Example 1: Preparation method, optimization of preparation conditions and identification of epidermal stem cells

[0043] 1. Preparation method of epidermal stem cells

[0044] At present, the preparation method of epidermal stem cells mainly includes two steps: 1) Firstly, a single skin epidermis is obtained through the functional separation of the junction between the epidermis and the dermis by neutral protease, which avoids the pollution of dermal cells; and, because trypsin damages the cells Therefore, the neutral protease digestion method is firstly used to separate the epidermal layer of the skin by using its function of hydrolyzing the junction between the epidermis and the dermis, and then the epidermal layer of the skin is digested with trypsin to obtain a single cell. It is beneficial to the protection of the activity of the digested cells and ensures that a sufficient number of active target cells are obtained; 2) the epidermal stem cells are then separated ...

Embodiment 2

[0099] Example 2: Preparation of bone marrow mesenchymal stem cells

[0100] The preparation of bone marrow mesenchymal stem cells can refer to the literature (Cai-Xia He, Ni Li, Yu-Lan Hu, Xiu-Mei Zhu, Hai-Jie Li, Min Han, Pei-Hong Miao, Zhong-Jie Hu, Gang Wang, Wen-Quan Liang, Yasuhiko Tabata, Jian-Qing Gao*, Effective gene delivery to mesenchymal stem cells based on the novel reverse transfection and three-dimensional cell culture system, Pharm Res, 2011, 28(7): 1577-1590)

[0101] 1) Preparation and in vitro culture of primary rat bone marrow mesenchymal stem cells (BMSCs): 3-week-old SD rats were sacrificed by neck pulling, the tibia and femur were separated, and the containing The tibia and femur were repeatedly washed with serum culture solution, and the obtained bone marrow was passed through a 200-mesh cell sieve (Shanghai Ruigu Biotechnology Co., Ltd.), centrifuged at 1000 rpm for 5 minutes, and the supernatant was discarded. After centrifugation, add the DMEM low-s...

Embodiment 3

[0104] Embodiment 3: the preparation of the medicine that accelerates skin layer repair and skin accessory organ regeneration

[0105] 1) The epidermal stem cells (ESCs) prepared by experimental group 7# in Example 1 were first washed twice with PBS, and then washed with trypsin-ethylenediaminetetraacetic acid (Trypsin-EDTA) containing 0.05% by weight of pancreatic Enzyme cell digestion solution (trypsin cell digestion solution obtained by dissolving trypsin-ethylenediaminetetraacetic acid in phosphate buffer at pH = 7.4) was digested for 7 minutes, and then added to the epidermal stem cells digested by trypsin cell digestion solution containing The DMEM low-sugar / F12 culture fluid of fetal bovine serum and penicillin streptomycin (the DMEM low-sugar / F12 culture fluid that contains fetal bovine serum and penicillin streptomycin is made of 10% fetal bovine serum by weight percentage, and the weight percentage is 1% penicillin, streptomycin, DMEM low-sugar culture medium of 59.3...

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