CN115671030A - Melanocyte-containing sodium hyaluronate hydrogel capable of being injected by water light needle, preparation method and application - Google Patents
Melanocyte-containing sodium hyaluronate hydrogel capable of being injected by water light needle, preparation method and application Download PDFInfo
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- CN115671030A CN115671030A CN202110822514.5A CN202110822514A CN115671030A CN 115671030 A CN115671030 A CN 115671030A CN 202110822514 A CN202110822514 A CN 202110822514A CN 115671030 A CN115671030 A CN 115671030A
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- sodium hyaluronate
- sodium
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- melanocytes
- water
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- 229920002385 Sodium hyaluronate Polymers 0.000 title claims abstract description 63
- 229940010747 sodium hyaluronate Drugs 0.000 title claims abstract description 63
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 title claims abstract description 63
- 239000000017 hydrogel Substances 0.000 title claims abstract description 48
- 210000002752 melanocyte Anatomy 0.000 title claims abstract description 46
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 238000000034 method Methods 0.000 claims abstract description 22
- 206010047642 Vitiligo Diseases 0.000 claims abstract description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 26
- 229960004393 lidocaine hydrochloride Drugs 0.000 claims description 15
- YECIFGHRMFEPJK-UHFFFAOYSA-N lidocaine hydrochloride monohydrate Chemical compound O.[Cl-].CC[NH+](CC)CC(=O)NC1=C(C)C=CC=C1C YECIFGHRMFEPJK-UHFFFAOYSA-N 0.000 claims description 15
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 13
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 13
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 13
- 239000011780 sodium chloride Substances 0.000 claims description 13
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 13
- 239000000243 solution Substances 0.000 claims description 10
- 230000003444 anaesthetic effect Effects 0.000 claims description 6
- 239000003431 cross linking reagent Substances 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- AFOSIXZFDONLBT-UHFFFAOYSA-N divinyl sulfone Chemical group C=CS(=O)(=O)C=C AFOSIXZFDONLBT-UHFFFAOYSA-N 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 4
- 238000002018 water-jet injection Methods 0.000 claims 1
- 238000002347 injection Methods 0.000 abstract description 7
- 239000007924 injection Substances 0.000 abstract description 7
- 239000003589 local anesthetic agent Substances 0.000 abstract description 4
- 230000006727 cell loss Effects 0.000 abstract description 2
- 230000000694 effects Effects 0.000 description 5
- 210000003491 skin Anatomy 0.000 description 5
- BLFLLBZGZJTVJG-UHFFFAOYSA-N benzocaine Chemical compound CCOC(=O)C1=CC=C(N)C=C1 BLFLLBZGZJTVJG-UHFFFAOYSA-N 0.000 description 4
- LEBVLXFERQHONN-UHFFFAOYSA-N 1-butyl-N-(2,6-dimethylphenyl)piperidine-2-carboxamide Chemical compound CCCCN1CCCCC1C(=O)NC1=C(C)C=CC=C1C LEBVLXFERQHONN-UHFFFAOYSA-N 0.000 description 2
- 206010024380 Leukoderma Diseases 0.000 description 2
- 102000015636 Oligopeptides Human genes 0.000 description 2
- 108010038807 Oligopeptides Proteins 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 229960005274 benzocaine Drugs 0.000 description 2
- 229960003150 bupivacaine Drugs 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 229960000385 dyclonine Drugs 0.000 description 2
- BZEWSEKUUPWQDQ-UHFFFAOYSA-N dyclonine Chemical compound C1=CC(OCCCC)=CC=C1C(=O)CCN1CCCCC1 BZEWSEKUUPWQDQ-UHFFFAOYSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000008506 pathogenesis Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 229960004919 procaine Drugs 0.000 description 2
- MFDFERRIHVXMIY-UHFFFAOYSA-N procaine Chemical compound CCN(CC)CCOC(=O)C1=CC=C(N)C=C1 MFDFERRIHVXMIY-UHFFFAOYSA-N 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010044278 Trace element deficiency Diseases 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000005784 autoimmunity Effects 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 238000013329 compounding Methods 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 210000003953 foreskin Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000037311 normal skin Effects 0.000 description 1
- 238000001126 phototherapy Methods 0.000 description 1
- 208000017520 skin disease Diseases 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
Landscapes
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention discloses a sodium hyaluronate hydrogel containing melanocytes, which can be injected by a water light needle, a preparation method and application. The sodium hyaluronate hydrogel prepared by the invention can be used as a carrier of melanocytes, so that cell loss is avoided, local anesthetic is added, and the operation is directly implemented in a water light injection mode, so that the pain caused by the operation is eliminated. The method also greatly simplifies the operation process of vitiligo treatment, and reduces the treatment cost.
Description
Technical Field
The invention belongs to the field of biomedicine, and particularly relates to a melanocyte-containing sodium hyaluronate hydrogel capable of being injected by a water light needle, a preparation method and application.
Background
Vitiligo is a relatively common acquired, generalized or localized depigmenting skin disease, mainly caused by the impairment or loss of melanocyte function. At present, the pathogenesis of vitiligo is not clear, but the pathogenesis is generally considered to be related to heredity, autoimmunity, inflammation induction, trauma, neuropsychiatric and human body trace element deficiency and the like. The main current methods for treating vitiligo include drug therapy, phototherapy and surgical therapy, which have large side effects and limited therapeutic effects, and surgical therapy is considered to be an effective method for treating vitiligo at present. The method of surgical treatment depends on the area of vitiligo skin. For patients with small vitiligo skin area, partial cure can be realized by surgical excision or skin grinding, and then melanocytes are transplanted, and for patients with large vitiligo skin area, autologous epidermal transplantation or melanocyte transplantation is required to achieve higher cure rate. Although the mode of surgical treatment achieves good effect, various disadvantages exist, such as causing new wound surface with appearance greatly different from normal skin, complicated process of transplanting melanocytes and easy loss or inactivation of cells, and the patients are painful and high in cost in the whole treatment process. Therefore, the development of a simple and effective vitiligo treatment method to make up for the problems existing in the prior art is a key problem to be solved at present.
Disclosure of Invention
The invention aims to solve the problems, and provides a melanocyte-containing sodium hyaluronate hydrogel capable of being injected by a water light needle, a preparation method and application thereof, so as to solve the problems of complex operation, pain of patients in the treatment process and high cost in the prior art.
In order to achieve the above object, the present invention adopts the following technical solutions:
a melanocyte-containing sodium hyaluronate hydrogel capable of being injected with a water needle, said hydrogel comprising melanocytes, sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and anesthetic.
Furthermore, the anesthetic is lidocaine hydrochloride, and the proportion range of the sodium hyaluronate, the disodium hydrogen phosphate, the sodium dihydrogen phosphate, the sodium chloride and the lidocaine hydrochloride is preferably (1% -5%), (0.5% -6.8%), (0.2% -3.3%), (0.8% -1%), (0.1% -0.5%); melanocyte content of 1 × 10 5 -1×10 8 One per mL.
The melanocyte-containing sodium hyaluronate hydrogel of the present invention may contain polypeptides, oligopeptides, amino acids or biologically active substances having an effect of promoting cell growth, or drugs having a local anesthetic effect such as procaine, benzocaine, bupivacaine, and dyclonine, in addition to the main components of melanocytes, sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride, and lidocaine hydrochloride.
In order to achieve the above object of the second aspect, the present invention adopts the following technical solutions:
the preparation method of the sodium hyaluronate hydrogel containing melanocytes and capable of being injected by a water light needle is characterized by comprising the following steps:
s1, dissolving sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and lidocaine hydrochloride in water to obtain a sodium hyaluronate solution;
s2, adding a cross-linking agent into the sodium hyaluronate solution, and stirring at room temperature to prepare sodium hyaluronate hydrogel;
and S3, inoculating melanocytes into the hydrogel to obtain the sodium hyaluronate hydrogel containing the melanocytes.
Further, the volume of the syringe is 0.1-5.0mL, and the diameter of the needle head of the water light needle ranges from 0.06mm to 0.11mm.
Further, the mass ratio of the sodium hyaluronate, the disodium hydrogen phosphate, the sodium dihydrogen phosphate, the sodium chloride and the lidocaine hydrochloride in the sodium hyaluronate hydrogel prepared in the step S1 is (1-5): (0.5-6.8): (0.2-3.3): (0.8-1): (0.1-0.5).
Further, the molecular weight range of the sodium hyaluronate in the step S1 is 150-400 ten thousand daltons; in the step S2, the cross-linking agent is divinyl sulfone, and the addition amount of the cross-linking agent is 0.05-0.25% of that of the sodium hyaluronate. The direct technical effect brought by the optimized scheme is that the prepared hydrogel is not too hard to inject, and is not easy to leak from an injection site during too soft injection, so that the operation is favorably implemented.
Further, the melanocyte concentration of the sodium hyaluronate hydrogel containing melanocytes in the sodium hyaluronate hydrogel prepared in the step S3 is 1 × 10 5 -1×10 8 one/mL.
The method of the invention prepares the sodium hyaluronate hydrogel containing melanocytes by compounding the melanocytes, sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and lidocaine hydrochloride, thereby avoiding loss of the melanocytes. The method can greatly simplify the operation process, eliminate the pain caused by the operation and reduce the cost generated in the treatment process by loading the sodium hyaluronate hydrogel containing melanocytes into an injector and injecting the hydrogel into the leukoderma of a patient with leucoderma by using a water-light instrument through a water-light needle.
In the preparation method, a polypeptide, an oligopeptide, an amino acid or a bioactive substance having a cell growth promoting effect may be seeded into the hydrogel together with melanocytes in the step S3, and a drug having a local anesthetic effect, such as procaine, benzocaine, bupivacaine, dyclonine, or the like, may be dissolved in water together with lidocaine hydrochloride in the step S1.
To achieve the object of the third aspect described above, namely: the invention relates to application of the melanocyte-containing sodium hyaluronate hydrogel in preparing a medicament for treating leucoderma. The medicine is suitable for injection of the water injection needle, the volume of the injector is 0.1-5.0mL, and the diameter range of the needle head of the water injection needle is 0.06-0.11mm.
The method for treating the leucoderma by injecting the sodium hyaluronate hydrogel containing the melanocyte with water light has the following advantages:
(1) According to the invention, firstly, the sodium hyaluronate hydrogel is prepared, then the melanocyte is inoculated into the hydrogel, and the obtained hydrogel can be used as a carrier of the melanocyte, so that cell loss is avoided.
(2) The invention directly injects the sodium hyaluronate gel containing melanocyte into the white spot skin of a patient with leucoderma by using the injector, the water light needle and the water light instrument, thereby greatly simplifying the operation process and reducing the treatment cost.
(3) According to the invention, a local anesthetic lidocaine hydrochloride is combined into the sodium hyaluronate hydrogel, and the operation is implemented in a water light injection mode, so that the pain caused by the operation is eliminated.
Detailed Description
Embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be readily apparent to those skilled in the art from the description herein. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
The following examples used the following starting materials:
sodium hyaluronate: the molecular weight range is 150-400 kilodaltons, and is available from Shandong Huaxi Biotech Ltd.
Disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride: from the national pharmaceutical group chemical agents limited.
Lidocaine hydrochloride: from Shanghai Aladdin Biotechnology Ltd.
Melanocytes: it is derived from foreskin of male children, and is given by children hospital in Hangzhou city.
Example 1
This example is a method for preparing a sodium hyaluronate hydrogel containing melanocytes, including the following steps:
s1, respectively weighing 2 parts of sodium hyaluronate, 0.9 part of disodium hydrogen phosphate, 0.4 part of sodium dihydrogen phosphate, 0.9 part of sodium chloride and 0.3 part of lidocaine hydrochloride according to parts by weight, and dissolving in 95.5 parts of purified water to obtain a sodium hyaluronate solution.
And S2, adding 0.1 part of divinyl sulfone into the sodium hyaluronate solution prepared in the S1, and stirring at room temperature to prepare the sodium hyaluronate hydrogel.
S3, taking 1mL of hydrogel prepared in S2, and adding 1X 10 5 And (4) obtaining the sodium hyaluronate hydrogel containing the melanocytes.
Example 2
The embodiment is a method for treating leucoderma by injecting sodium hyaluronate hydrogel containing melanocytes through water light, which comprises the following steps:
s1, respectively weighing 2 parts of sodium hyaluronate, 0.9 part of disodium hydrogen phosphate, 0.4 part of sodium dihydrogen phosphate, 0.9 part of sodium chloride and 0.3 part of lidocaine hydrochloride according to parts by mass, and dissolving in 95.5 parts of purified water to obtain a sodium hyaluronate solution.
And S2, adding 0.1 part of divinyl sulfone into the sodium hyaluronate solution prepared in the S1, and stirring at room temperature to prepare the sodium hyaluronate hydrogel.
S3, taking 1mL of hydrogel prepared in S2, and adding 1X 10 6 And (4) obtaining the sodium hyaluronate hydrogel containing the melanocytes.
And S4, filling the sodium hyaluronate hydrogel containing melanocytes obtained in the step S3 into a 2mL syringe, and injecting the hydrogel into leukoderma of a patient with leucoderma by using a water light instrument through a water light needle with the needle head diameter of 0.06 mm.
Wherein, the injector (2 mL Shanghai Bidi medical apparatus Co., ltd., china) and the water light instrument (V)ital Injector2 TM Beijing di ke excellence science and technology Co., ltd., china), shuizhouguang needle (32G, hangzhou cruise medicine Co., ltd., china).
Although the invention has been described in detail above with reference to a general description and specific examples, it will be apparent to one skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, it is intended that all such modifications and variations be included within the scope of the invention as claimed and not departing from the spirit thereof.
Claims (10)
1. A melanocyte-containing sodium hyaluronate hydrogel capable of being injected with a water needle, said hydrogel comprising melanocytes, sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and anesthetic.
2. The sodium hyaluronate hydrogel containing melanocytes capable of being injected with an aqueous light needle according to claim 1, wherein the anesthetic is lidocaine hydrochloride, and the ratio range of sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and lidocaine hydrochloride is preferably (1% -5%) (0.5% -6.8%) (0.2% -3.3%) (0.8% -1%) (0.1% -0.5%); melanocyte content of 1 × 10 5 -1×10 8 one/mL.
3. The preparation method of the sodium hyaluronate hydrogel containing melanocytes and capable of being injected by a water light needle is characterized by comprising the following steps:
s1, dissolving sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and lidocaine hydrochloride in water to obtain a sodium hyaluronate solution;
s2, adding a cross-linking agent into the sodium hyaluronate solution, and stirring at room temperature to prepare sodium hyaluronate hydrogel;
and S3, inoculating melanocytes into the hydrogel to obtain the sodium hyaluronate hydrogel containing the melanocytes.
4. The method according to claim 3, wherein the sodium hyaluronate used has a molecular weight ranging from 150 to 400 ten thousand daltons.
5. The method of claim 3, wherein the syringe has a capacity of 0.1-5.0mL and the water light needle has a needle diameter in the range of 0.06-0.11mm.
6. The method according to claim 3, wherein the sodium hyaluronate hydrogel prepared in step S1 comprises sodium hyaluronate, disodium hydrogen phosphate, sodium dihydrogen phosphate, sodium chloride and lidocaine hydrochloride in a mass ratio of (1-5): 0.5-6.8): 0.2-3.3): 0.8-1): 0.1-0.5.
7. The method according to claim 4, wherein the molecular weight of the sodium hyaluronate in the step S1 is in the range of 150 to 400 ten thousand daltons; in the step S2, the cross-linking agent is divinyl sulfone, and the addition amount of the cross-linking agent is 0.05-0.25% of that of the sodium hyaluronate.
8. The method according to claim 4, wherein the sodium hyaluronate hydrogel containing melanocytes in the sodium hyaluronate hydrogel prepared in step S3 has a melanocyte concentration of 1X 10 5 -1×10 8 one/mL.
9. Use of the sodium hyaluronate hydrogel containing melanocytes injectable with aqueous light needle according to claim 1 or 2 for the preparation of a medicament for the treatment of vitiligo.
10. The use of claim 9, wherein the medicament is suitable for water-jet injection, the syringe has a volume of 0.1-5.0mL and the water-jet has a needle diameter in the range of 0.06-0.11mm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202110822514.5A CN115671030A (en) | 2021-07-21 | 2021-07-21 | Melanocyte-containing sodium hyaluronate hydrogel capable of being injected by water light needle, preparation method and application |
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| CN202110822514.5A CN115671030A (en) | 2021-07-21 | 2021-07-21 | Melanocyte-containing sodium hyaluronate hydrogel capable of being injected by water light needle, preparation method and application |
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