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Dopamine-hyaluronic acid coating material, preparation method and its application in promoting specific adhesion of dental pulp stem cells to dentin

A technology of dental pulp stem cells and hyaluronic acid, which is applied in the directions of hyaluronic acid coating, chitin polysaccharide coating, coating, etc. The odontoblast layer and dentin are poorly formed, and the regeneration effect of the pulp-dentin complex is not good, so as to avoid toxic side effects, good biocompatibility, and easy operation.

Active Publication Date: 2019-12-24
JILIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, some researchers have used tissue cell homing (Cell homing) or transplanted human stem cells from human exfoliated deciduous (SHED), dental pulp stem cells (Dental pulp stem cells, DPSC), human umbilical vein endothelial cells (Human umbilical vein endothelial cells) endothelial cells, HUVEC) as seed cells, growth factors such as TGF-β1, BMP-2, bFGF, VEGF, PDGF, NGF and BMP7, and platelet-rich plasma (Platelet–rich plasma, PRP), silk protein, PLLA microspheres , hyaluronic acid gel or collagen gel, etc. were used as scaffolds to study the pulp regeneration of full-length dental roots (or fragments), which can form pulp-like tissue and differentiate into odontoblasts. However, the odontoblast layer and The formation of dentin is poor, the regeneration effect of the pulp-dentin complex is not good, and the regenerated pulp-like tissue, especially the odontoblasts differentiated from seed cells such as DPSC, is not firmly combined with the dentin of the root canal wall, which ultimately hinders the regeneration of the dental pulp. regenerative development

Method used

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  • Dopamine-hyaluronic acid coating material, preparation method and its application in promoting specific adhesion of dental pulp stem cells to dentin
  • Dopamine-hyaluronic acid coating material, preparation method and its application in promoting specific adhesion of dental pulp stem cells to dentin
  • Dopamine-hyaluronic acid coating material, preparation method and its application in promoting specific adhesion of dental pulp stem cells to dentin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028]Taking the molar ratio of dopamine hydrochloride and hyaluronic acid as 1:1 as an example, 201.6 mg of hyaluronic acid was dissolved in 50 mL of deionized water, 191.7 mg of EDC and 115.09 mg of NHS were slowly added to the solution, stirred evenly for 20 min, and then added to Add 94.82 mg of dopamine hydrochloride to the mixed solution. The reaction solution was protected under nitrogen and stirred at room temperature, adjusted to pH 4 with hydrochloric acid, kept for 3 hours, and then continued to react overnight at room temperature. After the reaction, the solution was purified by dialysis (molecular weight cut-off = 12000Da) in an acidic environment of pH = 5 deionized water for 4 days, and dried in vacuum freezer (-70°C) for 72 hours to obtain a white flocculent graft product called dopamine-hyalurin Acid coating material, product mass 234.16mg. Change the feed ratio of dopamine hydrochloride and hyaluronic acid, and prepare coating materials under other condition...

Embodiment 2

[0033] Preparation of dentin tissue blocks: freshly extracted premolars for orthodontic treatment were fully sterilized with 2-5% sodium hypochlorite aqueous solution, and rinsed with PBS (phosphate buffered saline); then, the periodontal tissue was removed and applied PBS was cooled down to obtain the tooth root, and the pulp and predentin were completely removed, and the dentine tissue blocks of 3.0mm*3.0mm*1.0mm were cut under water spray cooling; all the dentine tissue blocks were soaked in deionized water for 5 hours in turn, Ultrasonic cleaning for 20 minutes, soaking in 17% EDTA aqueous solution for 101 minutes, soaking in 19% citric acid aqueous solution for 1 minute, and then disinfecting with 5.25% sodium hypochlorite aqueous solution for 30 minutes, then rinsing and soaking in PBS for 2 days, to obtain dentin tissue blocks.

[0034] The dopamine-hyaluronic acid coating material with a molar ratio of 1:1 was dissolved in 10 mL of deionized water at 25 mg, 50 mg and 10...

Embodiment 3

[0038] Select a dopamine-hyaluronic acid solution with a molar ratio of 5:1 and a concentration of 2.5 mg / mL, 5 mg / mL, and 10 mg / mL, and immerse the dentin tissue pieces prepared in Example 2 into the above solution, and take them out after 24 hours. Spin coating with a homogenizer (400r / s; 20s), and then transfer to a vacuum desiccator for freeze-drying for 48h. SEM observation of the micro-morphological characteristics of the dentin surface coating. The experimental data are listed in Table 3.

[0039] Table 3: Dentin coating data prepared with different concentrations of dopamine-hyaluronic acid solution at a molar ratio of 5:1

[0040]

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Abstract

The invention discloses a dopamine-hyaluronic acid coating material as well as a preparation method and application thereof in promoting specific adhesion of dental pulp stem cell to dentin, and belongs to the technical field of dental pulp regeneration. The preparation method comprises the following steps: sequentially adding raw materials including hyaluronic acid, 1-(3-dimethylamino propyl)-3-ethyl carbodiimide, N-hydroxyl succinimide and dopamine hydrochloride into deionized water according to a molar ratio of (1 to 10) to 2 to 2 to 1, and mixing uniformly; stirring at room temperature under nitrogen protection; adjusting the pH to 4-5 with hydrochloric acid or sodium hydroxide; reacting for 2-5 hours while keeping the pH value unchanged; reacting overnight at room temperature; finally, performing dialysis and purification on the obtained reaction solution for 3-5 days in an acid deionized water environment; collecting the product; performing vacuum freeze drying for 48-96h, wherein the obtained white flocculent grafted product is a dopamine-hyaluronic acid composite material. The coating thickness is controlled by changing the molar ratio of the raw materials as well as the product concentration, and the coating material can effectively promote specific adhesion of dental pulp stem cell to the root canal wall.

Description

technical field [0001] The invention belongs to the technical field of dental pulp regeneration, and in particular relates to a dopamine-hyaluronic acid coating material, a preparation method and its application in promoting specific adhesion of dental pulp stem cells to dentin. Background technique [0002] Pulp infection, trauma, and certain maxillofacial surgeries require the removal of infected pulp tissue, and the loss of living pulp leads to paresthesia, tooth discoloration, brittleness, and tooth splitting due to loss of nutrition in the hard tissue of the tooth , or even fracture or tooth extraction, etc., the complications of traditional endodontic treatment seriously endanger the patient's oral function, appearance and physical and mental health. Therefore, pulp regeneration has become a pulp therapy method with bright application prospects. Many scholars are competing to devote themselves to the research on pulp regeneration, and it has also become a research hots...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09D105/08C09D7/63A61L27/28A61L27/34A61L27/50A61L27/58
CPCA61L27/28A61L27/34A61L27/50A61L27/58A61L2400/18A61L2420/02A61L2420/06A61L2430/12C08K5/18C08K5/29C08K5/3415C09D105/08C08L5/08
Inventor 李祥伟刘亚楠宋文龙张雪巍姜钧天杨柏
Owner JILIN UNIV
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