CN111407755A - Use of retinoic acid in tooth development - Google Patents
Use of retinoic acid in tooth development Download PDFInfo
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- CN111407755A CN111407755A CN202010366054.5A CN202010366054A CN111407755A CN 111407755 A CN111407755 A CN 111407755A CN 202010366054 A CN202010366054 A CN 202010366054A CN 111407755 A CN111407755 A CN 111407755A
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- retinoic acid
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- teeth
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Images
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
- A61K31/203—Retinoic acids ; Salts thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/13—Amines
- A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
- A61K31/137—Arylalkylamines, e.g. amphetamine, epinephrine, salbutamol, ephedrine or methadone
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
Abstract
The invention relates to the field of biomedicine, and discloses application of retinoic acid and a retinoic acid inhibitor in preparation of a tooth treatment drug, wherein the retinoic acid can promote pharyngeal tooth mineralization, and the retinoic acid inhibitor can inhibit proliferation and migration of cranial nerve spine cells, so that regulation and control of early development of teeth are realized. The retinoic acid and the inhibitor thereof can be used as a novel tooth regeneration drug to be applied to clinical practice.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to application of retinoic acid in preparation of a tooth regeneration medicine.
Background
Teeth are the hardest organs of the human body and are also organs that cannot be regenerated after injury. Because the teeth are positioned at the most front end of the digestive tract, the chewing and grinding of food are beneficial to further digestion and absorption of food, and once the teeth are damaged, the nutrient intake is greatly influenced, so that the normal operation of the organs of the whole body is influenced. Because the loss of teeth caused by decayed teeth, periodontitis, trauma and the like can seriously reduce the life quality of people and even influence the health of the whole body, the tooth damage repair is a topic which is always researched by researchers. At present, the damage restoration of teeth mainly depends on filling and covering of biological materials such as resin and the like and planting and restoration after tooth falling, the restoration methods can quickly restore the tooth functions, but have some defects, such as high requirements on the oral environment, large technical sensitivity difference of clinical workers and other factors, which can cause uneven restoration effect, poor marginal sealing performance of restoration, loosening and falling and the like, and further aggravate the adverse consequences of tooth damage, oral chronic inflammation and the like. Therefore, the search for a safer and more reliable method of tooth loss restoration-tooth regeneration is the direction of all oral workers.
Disclosure of Invention
The invention aims to provide application of retinoic acid in preparation of a tooth regeneration medicine to realize tooth regeneration and solve the problems that the tooth injury and oral chronic inflammation are further aggravated by poor sealing performance of a restoration edge, loosening and falling and the like in a conventional restoration method in the prior art.
In order to achieve the purpose, the invention adopts the following technical scheme: use of retinoic acid in the preparation of a medicament for tooth regeneration.
The principle and the advantages of the scheme are as follows: retinoic acid is the main active metabolite of vitamin a, and its synthesis is mainly characterized by that the alcohol mode retinol of vitamin a is oxidized into retinaldehyde by retinol dehydrogenase, and then the retinaldehyde is oxidized into retinoic acid by retinaldehyde dehydrogenase; degraded by cytochrome P450 isoenzymes; retinoic acid functions by binding to two nuclear receptors RAR, RXR, which form heterodimers that bind to DNA at the Retinoic Acid Reaction Element (RARE). When the organ development does not need retinoic acid signals to participate, RAR and RXR dimers recruit co-inhibitors to inhibit the expression of target genes, and when the retinoic acid signals are sufficient again, receptor dimers are depolymerized with inhibitors to combine with retinoic acid to promote the transcription of downstream target genes so as to regulate the development of various organs.
The inventor finds that the retinoic acid signal can influence the gene expression related to tooth regeneration, mainly influences the mineralization of teeth and is realized by regulating the proliferation and migration of cranial nerve spine cells by accident in the process of researching the influence of retinoic acid on tooth development. The inventor utilizes in-situ hybridization, alizarin red staining, confocal microscope detection to construct over-expression fish of retinoic acid degrading enzyme and synthetase, utilizes a specific injury and embryo heat shock mode to verify and evaluate the activity of retinoic acid on tooth mineralization and proliferation and migration of cranial nerve spine cells, discovers a new effect of retinoic acid, widens the application range of retinoic acid, and provides a theoretical basis for research and development of tooth regeneration and repair medicines.
Preferably, as an improvement, the tooth-regenerating drug further comprises a retinoic acid inhibitor, and the retinoic acid inhibitor acts after the retinoic acid.
By adopting the technical scheme, the applicant accidentally discovers in the research process that the expression detection result of the specific marker gene of the pharyngeal teeth shows that the expression of the gene in the retinoic acid treatment group is slightly stronger than that in the control group, and no related gene expression exists in the inhibitor treatment group, so that the influence of the function weakening on the development of the pharyngeal teeth is larger compared with the function strengthening, and the retinoic acid inhibitor influences the subsequent development of the teeth by weakening the expression of the specific marker gene of the pharyngeal teeth after the teeth are mineralized. The aim of promoting the development and regeneration of teeth is achieved by sequentially exerting the effects of retinoic acid and a retinoic acid inhibitor.
By adopting the technical scheme, the method for exploring the regulation and control effect of retinoic acid on the tooth development process by using exogenous all-trans retinoic acid is convenient, rapid and easy to operate. Compared with other endogenous exploration means such as constructing transgenic animals and the like, the method has short required time, and the exogenous retinoic acid is safe and nontoxic and can be directly purchased from biological companies.
By adopting the technical scheme, the diethylaminobenzaldehyde is used for inhibiting retinoic acid signals, so that the regulation and control effect of retinoic acid on the tooth development process is explored, and the method is convenient, rapid and easy to operate. Compared with other endogenous exploration means such as constructing transgenic animals and gene knockout, the method has the advantages that the required time is short, the treatment solution with proper concentration can be prepared according to actual needs so as to achieve the inhibition effect on retinoic acid signals in different degrees, and the method can be directly purchased from biological companies, and is convenient and easy to operate.
Preferably, as an improvement, the teeth are pharyngeal teeth, and the retinoic acid and the inhibitor thereof are prepared into pharmaceutically acceptable dosage forms according to the conventional pharmaceutical preparation method.
By adopting the technical scheme, the pharyngeal teeth are also called gill teeth, hypopharynx teeth and throat teeth. Teleost fish grow on the teeth of the gill arch. In the developed species of pharyngeal teeth, the jaw teeth are not developed, whereas in the developed species of jaw teeth, the pharyngeal teeth are poorly developed or not developed. The tested objects are the pharyngeal teeth of the zebra fish, are representative, and can provide reference for the influence of follow-up research on the full teeth. The retinoic acid and the inhibitor thereof are prepared into different dosage forms, so that a proper pharmaceutical dosage form can be reasonably selected according to different administration modes, the application range is wide, and the selectable space is large.
Preferably, as an improvement, retinoic acid is used as a pharyngeal tooth mineralization promoter.
By adopting the technical scheme, the influence of retinoic acid on tooth regeneration is to improve tooth regeneration capacity by promoting tooth mineralization, and the inventor shows that the mineralization of the dental pharyngeal teeth of the zebra fish treated by the retinoic acid is enhanced through an alizarin red staining experiment result.
Preferably, as an improvement, the retinoic acid inhibitor is used as a cell migration inhibitor for cranial nerve spine.
By adopting the technical scheme, the migration of the cranial nerve spine cells is in important connection with the development of teeth, after the treatment of the retinoic acid inhibitor, the expression of a gene snail1a related to the epithelial-mesenchymal transition process of the cranial nerve spine cells has no obvious change, and the expression of a gene cad related to the migration of the cranial nerve spine cells is weakened, namely, the retinoic acid inhibitor has the function of inhibiting the migration of the cranial nerve spine cells and can be used as the cranial nerve spine cell migration inhibitor to realize the regulation and control of the early development of teeth.
Preferably, as an improvement, the retinoic acid inhibitor is used as a cell proliferation inhibitor of cranial nerve spine.
By adopting the technical scheme, the migration of the cranial nerve spine cells is in important connection with the development of teeth, after the treatment of the retinoic acid inhibitor, the expression of a gene snail1a related to the epithelial-mesenchymal transition process of the cranial nerve spine cells has no obvious change, and the expression of a gene cad related to the proliferation of the cranial nerve spine cells is weakened, namely, the retinoic acid inhibitor has the function of inhibiting the proliferation of the cranial nerve spine cells and can be used as the cranial nerve spine cell proliferation inhibitor to realize the regulation and control of the early development of teeth.
Preferably, the concentration of retinoic acid promoting pharyngeal tooth mineralization is more than or equal to 3 × 10-7M。
With the above technical solution, 3 × 10 is used-7And (3) after retinoic acid treatment of M, collecting 5dpf embryos, fixing, and then carrying out alizarin red staining, so that pharyngeal tooth mineralization is obviously enhanced.
Preferably, as an improvement, the concentration of retinoic acid inhibitor for inhibiting migration and proliferation of cranial nerve spine cells is 0.5 × 10-7M-1×10-7M。
By adopting the technical scheme, when the concentration of the retinoic acid inhibitor is 0.5 × 10-7M-1×10-7In M, in-situ hybridization detection results of embryos after the zebra fish is treated by the retinoic acid inhibitor show that 48hpf and 56hpf have no obvious difference in expression of posterior and lateral genes snail1a, and only a slight reduction in expression is observed on the 72hpf side; whereas cad expression was significantly attenuated after 48 hpf.
Drawings
FIG. 1 shows the results of in situ hybridization of gene pitx2 after treatment with RA and DEAB according to example 1 of the present invention;
FIG. 2 shows the results of in situ hybridization of gene fth1b after treatment with RA and DEAB in example 1 of the present invention;
FIG. 3 shows the results of alizarin red staining at 5dpf after RA and DEAB treatment in example 1 of the present invention;
FIG. 4 is a diagram showing an expression pattern of snail1a after DEAB treatment in example 2 of the present invention;
FIG. 5 is a graph showing the expression pattern of cad after DEAB treatment in example 2 of the present invention;
FIG. 6 shows the results of staining with alizarin red for 48h after regeneration of zebrafish Tg (dlx2b: Dendra 2-NTR; hsp: aldh1a2-GFP) (a) and Tg (dlx2b: Dendra 2-NTR; hsp: cyp26b1-GFP) (b) after pharyngeal tooth injury in example 3 of the present invention;
FIG. 7 shows the results of confocal microscope observations of regeneration of 24h and 48h after pharyngeal tooth injury in zebrafish Tg (dlx2b: Dendra 2-NTR; hsp: aldh1a2-GFP) and Tg (dlx2b: Dendra 2-NTR; hsp: cyp26b1-GFP) according to example 3 of the invention.
Detailed Description
Description of the reagents:
example 1: growth condition of pharyngolaryngeal teeth of zebra fish treated by exogenous retinoic acid and inhibitor thereof
Experimental animals: all zebra fish were bred and bred strictly following standard experimental conditions as specified by the regulations of the animal care committee for laboratory use, using zebra fish as model animal. The zebra fish embryos used in the experiment were soaked in 0.003% PTU to inhibit melanin production for easy observation.
Experimental reagent: the reagents used in the experiment include all-trans Retinoic Acid (RA), diethylamino benzaldehyde (DEAB), in-situ hybridization related reagent (Roche), developing solution (Roche) and 0.5% alizarin red staining solution.
The experimental method comprises dissolving RA and DEAB powder in 100% DMSO respectively to obtain final concentration of 1 × 10-3M stock was stored at-20 ℃ for embryo handling 3. mu.l DEAB and 9ul RA stock were added to 30ml egg water to a final concentration of 1 × 10-7M and 3 × 10-7M, soaking the embryo in the treating solution, wherein the treatment time of the medicine is 24-36 hpf. Control embryos were treated with 0.2% DMSO.
Whole embryo in situ hybridization:
(1) drug-treated 48hpf, 72hpf embryos and DMSO control 48hpf, 72hpf embryos were collected, 20 embryos per tube, fixed with 4% PFA overnight at 4 ℃.
(2) The collected embryos were dehydrated and rinsed 5 times for 30min each with 100% absolute methanol, and the dehydrated embryos were left overnight at-20 ℃.
(3) Taking out the embryo, rehydrating, sequentially rinsing with 75% anhydrous methanol/25% 1 × PBT, 50% anhydrous methanol/50% 1 × PBT, 25% anhydrous methanol/75% 1 × PBT, and 100% 1 × PBT, and rinsing 1ml solution each time at room temperature for 5 min.
(4) Embryo digestion, 1ml of 100% 1 × PBT is rinsed for 5min, proteinase K is prepared into a digestion solution with a final concentration of 10ug/ml by using a 1 × PBT solution, 1ml of the digestion solution is added into each tube, and the corresponding period is slowly treated on a flat shaking bed according to the embryo period (48hpf embryo digestion is 20min, 56hpf embryo digestion is 25min, and 72hpf embryo digestion is 30 min).
(5)1ml of 1 × PBT solution was rinsed twice for 5min each, the embryos were washed free of residual proteinase K, 1ml of 4% PFA fixed embryos for 30min, and 1 × PBT was rinsed 5 times for 5min each with 1 ml.
(6) The Hyb buffer was preheated in advance, 1ml of preheated Hyb buffer was added after 1ml of 50% 1 × PBT/50% Hyb buffer was used for embryo rinsing, and prehybridization was carried out in a water bath at 68.5 ℃ for 3-5 h.
(7) The Hyb buffer was removed and the pre-warmed probe hybridization solution (3. mu.l probe was added to 200. mu.l Hyb buffer) was added, 200. mu.l per tube, overnight in a water bath at 68.5 ℃.
(8) The next day the probes were recovered and stored at-20 ℃.
(9) Embryos were rinsed in a 68.5 ℃ water bath with pre-warmed rinsing solution of 100% Hybbuffer, 25% 2 × SSCT/75% Hyb buffer, 50% 2 × SSCT/50% Hyb buffer, 75% 2 × SSCT/25% Hyb buffer, 100% 2 × SSCT in sequence, each buffer was rinsed for 10min, followed by 4 rinses with pre-warmed 0.2 × SSCT, 15min each.
(10) The embryos are rinsed at room temperature, the rinsing solutions are 75% 0.2 × SSCT/25% 1 × MABT, 50% 0.2 × SSCT/50% 1 × MABT, 25% 0.2 × SSCT/75% 1 × MABT and 100% 1 × MABT in sequence, and the embryos are rinsed once and 1ml each time for 5 min.
(11) Adding 1ml of 1 × blocking solution into each tube of the rinsed embryos, sealing the tubes on a shaking table at normal temperature for 2h, (12) removing the 1 × blocking solution, preparing the 1 × blocking solution and the anti-Dig-AP antibody according to the ratio of 1: 1000, adding 200 mu l into each tube, and incubating overnight on the shaking table at 4 ℃.
(13) The second antibody is recovered on the third day and stored at 4 ℃ for two times.
(14)1ml of 1 × MABT solution rinse embryos 8 times, 15min each time (15) freshly prepared NTMT solution rinse embryos 3 times, 5min each time, after which the embryos are transferred to 24-well plates and each embryo marker is done.
(16) And (3) adding 20 mul of NBT/BCIP into 1ml of NTMT solution to prepare a developing solution, adding 500 mul of developing solution into each hole, developing in a dark place at 37 ℃, and observing the developing condition under a microscope every 20min according to the working condition of the probe.
(17) When no great difference is found in the embryo development state through two successive observations, the development can be stopped, the development solution is removed, 1ml of stop solution is rinsed for 3 times, each time for 5min, and then the images can be collected at 4 ℃ at a selected period.
Alizarin red staining:
(1) one tube of each of the drug-treated and control embryos at 5dpf was collected, 20 per tube, and 1ml of 4% PFA was fixed overnight at 4 ℃.
(2) The 1 × PBST solution was rinsed 2 times for 5min each, after which embryonic heart and liver tissue were dissected under a microscope to expose pharyngeal teeth thoroughly.
(3) Embryos were dehydrated for 30min with 50% ethanol at room temperature.
(4) 1ml of staining solution 1ml of 1 × PBST/20ul of 0.05% alizarin red was added for overnight staining at room temperature in the dark.
After 24hpf-36hpf zebrafish embryos are treated by RA and DEAB, 48hpf, 56hpf and 72hpf embryos are collected after the treatment is stopped, fixed and dehydrated and used for detecting whole embryo in situ hybridization, and probes selected by people herein are zebrafish pharyngeal tooth specific marker genes pitx2 and fth1b, the probes are provided by Zhengxuedan, and the probes are proved to work normally.
As shown by the in situ hybridization results in FIGS. 1 and 2, the hybridization was performed at 3 × 10-7In M RA treatment group, at 48hpf and 72hpf, the expression of the pharyngeal tooth marker gene pitx2 was not significantly different from the control group, but at 1 × 10-7No expression of pitx2 was observed for 48hpf and 72hpf in M DEAB-treated group, 3 × 10-7In M RA-treated group, expression of fth1b was not significantly different from that in control group at 56hpf 72hpf, 1 × 10-7In the M DEAB-treated group, 56hp and 72hpf were not expressed in fth1 b.
As shown in FIG. 3, 5dpf embryos were collected after drug treatment of embryos at 24-36 hpf and fixed for alizarin red staining. Alizarin red staining mainly stains mineralized tissues including teeth, bones and the like. From the alizarin red staining result, it was found that in the control group, stronger 4V was seen1Mineralisation and weaker 5V1Dental apices mineralized at 3 × 10-7The mineralization of the pharyngeal teeth of the zebra fish is stronger in the M RA treatment group than in the control group, and the RA group is 4V1Has strong mineralization and can be seen in 3V1And 5V1Stronger dental tip mineralization than control group, 1 × 10-7The MDEAB treated group did not find the presence of mineralized pharyngeal teeth.
The above results illustrate that: retinoic acid can affect pharyngeal tooth development by affecting the expression of pitx2 and fth1b genes, and retinoic acid signaling can promote pharyngeal tooth mineralization.
Example 2: gene expression change related to neural spine cells after retinoic acid signal inhibition
TRIZO L (L if Technologies), chloroform (Sangon), isopropanol (Sangon), reverse transcription Kit (Roche), rTaq DNA polymerase (TaKaRa), pGEM @ T Easy Vector Kit (Promega), DH5 α allelochemicals (Invitrogen), agrose (Invitrogen), Gel extraction Kit (Qiagen), DIG RNA labeling Kit (Roche), T7RNA polymerase (Roche), DNase I enzyme (New England Biolabs), and the rest of the reagents were the same as in example 1.
Extracting Total RNA of zebra fish embryos, synthesizing cDNA and a probe, purifying the probe, performing an in situ hybridization experiment, treating the zebra fish embryos with 24hpf-36hpf by using a retinoic acid signal inhibitor, and collecting embryos of 48hpf, 56hpf and 72hpf to detect the expression changes of snail1a and cad.
As shown in FIGS. 4 and 5, in situ hybridization of 48hpf, 56hpf and 72hpf embryos after drug treatment revealed no significant difference in expression of the dorsal and lateral genes, snail1a, from the control group, and only a slight decrease in expression was observed at the 72hpf side; the expression of the gene cad is obviously different between the drug treatment group and the control group, the cad expression of the experimental group is obviously weaker than that of the control group at 48hpf, the cad expression difference in the two groups is smaller and smaller along with the proliferation and migration of cranial nerve spine cells to corresponding parts, but the cad expression of the experimental group is weaker than that of the control group as a whole.
The above results illustrate that: the expression of a gene snail1a related to the epithelial-mesenchymal transition process of cranial nerve spine cells has no obvious change, while the expression of a gene cad related to the proliferation and migration of cranial nerve spine cells is obviously weakened in a drug treatment group, namely, the retinoic acid signal controls the early development of the pharyngeal teeth of the zebra fish to play a role mainly by influencing the proliferation and migration of the cranial nerve spine cells.
Example 3: effect of retinoic acid signal inhibition on regeneration of pharyngolaryngitis of zebrafish
Constructing retinoic acid signal synthetase and degrading enzyme overexpression fish: cloning CDS fragments of cyp26b1 and aldh1a2 from a zebra fish genome, and selecting a non-mutation plasmid and hsp70l- (-) -gfp by plasmid screening and sequencing after TA cloning; the cryaa-cerulean subclones successfully constructed recombinant plasmids hsp: cyp26b1-GFP and hsp: aldh1a 2-GFP. After being injected into ABGO background zebra fish single-cell embryos in a microscopic mode, the zebra fish embryos are subjected to heat shock at 3dpf, and green fluorescent protein is expressed in the whole body, namely, overexpression fish hsp: cyp26b1-GFP and hsp: aldh1a2-GFP are constructed successfully. The gene can be stably inherited after three successive generations of forward genetic screening to F3 generation.
MTZ treated zebrafish embryos: hybridizing transgenic zebrafish Tg (dlx2b: Dendra2-NTR) with hsp: cyp26b1-GFP and hsp: aldh1a2-GFP respectively, treating the hybridized embryos with 12mM MTZ solution, soaking 3dpf zebrafish embryos in the MTZ solution for 48hpf, stopping MTZ treatment at 5dpf, thermally shocking the embryos for 30min at 38.5 ℃, recovering for 24h, and observing the regeneration difference of pharyngeal teeth of the experimental group and the control group after recovering for 48 h.
The rest of the process is the same as in example 1.
Pharyngeal tooth48h after injury regeneration, collecting embryos, and observing pharyngeal tooth regeneration conditions by alizarin red staining, wherein the result is shown in figure 6, in Tg (dlx2b: Dendra 2-NTR; hsp: aldh1a2-GFP), the mineralization of regenerated pharyngeal teeth is obviously stronger than that of a control group after the injured pharyngeal teeth of zebra fish, and the regenerated pharyngeal teeth can be observed in the alizarin red staining result, and the 4V strong in the control group can be seen1Mineralisation and weaker 3V1And 5V1Dental tip mineralization, 3V in the experimental group1、4V1And 5V1The mineralization is stronger than that of the control group; whereas in Tg (dlx2b: Dendra 2-NTR; hsp: cyp26b1-GFP) the mineralization of the regenerated pharyngeal teeth is weaker than that of the control group, and alizarin red staining results can be observed, in which the stronger 4V is seen in the control group1Mineralisation and weaker 3V1And 5V1Mineralization whereas in the experimental group, 3V1And 5V1Mineralization invisible and 4V1Mineralization was also weaker than control.
After regeneration of pharyngeal teeth, embryos are collected for 24h (R24h) and 48h (R48h) respectively, the pharyngeal teeth are observed under a confocal microscope after the pharyngeal teeth are peeled off and completely exposed by a body type microscope, the result is shown in figure 7, the observation result of the confocal microscope is consistent with the alizarin red staining result, and stronger mineralized 4V can be seen in Tg (dlx2b: Dendra 2-NTR; hsp: aldh1a2-GFP)1，3V1And 5V1Is also stronger than the control, whereas in Tg (dlx2b: Dendra 2-NTR; hsp: cyp26b1-GFP) 4V1Weakly mineralized, 3V1And 5V1It can be seen that the tooth germ has not been mineralized, and the secondary result is consistent with the alizarin red staining result.
The above results illustrate that: the retinoic acid signal influences the mineralization of regenerated pharyngeal teeth in the regeneration process of the zebra fish pharyngeal teeth, and does not influence the number and the shape of the regenerated pharyngeal teeth.
The foregoing is merely an example of the present invention and common general knowledge in the art of designing and/or characterizing particular aspects and/or features is not described in any greater detail herein. It should be noted that, for those skilled in the art, without departing from the technical solution of the present invention, several variations and modifications can be made, which should also be regarded as the protection scope of the present invention, and these will not affect the effect of the implementation of the present invention and the practicability of the patent. The scope of the claims of the present application shall be determined by the contents of the claims, and the description of the embodiments and the like in the specification shall be used to explain the contents of the claims.
Claims (10)
1. Use of retinoic acid in the preparation of a medicament for tooth regeneration.
2. Use of retinoic acid in the preparation of a tooth-regenerating medicament according to claim 1, wherein: the tooth regeneration medicine also comprises retinoic acid inhibitor, and the retinoic acid inhibitor acts after retinoic acid.
3. Use of retinoic acid in the preparation of a tooth-regenerating medicament according to claim 1, wherein: the retinoic acid is exogenous all-trans retinoic acid, and the molecular formula of the all-trans retinoic acid is as follows: c20H28O2The structural formula is as follows:
4. use of retinoic acid in the preparation of a tooth-regenerating medicament according to claim 2, wherein: the retinoic acid inhibitor is diethylaminobenzaldehyde, and the molecular formula of the diethylaminobenzaldehyde is C11H15NO, structural formula:
5. use of retinoic acid in the preparation of a tooth-regenerating medicament according to claim 2, wherein: the teeth are pharyngeal teeth, and the retinoic acid and the inhibitor thereof are prepared into pharmaceutically acceptable dosage forms according to the conventional pharmaceutical preparation method.
6. Application of retinoic acid as pharynx and dental mineralization promoter is provided.
7. Application of retinoic acid inhibitor as cranial nerve spine cell migration inhibitor is provided.
8. Application of retinoic acid inhibitor as cranial nerve spine cell proliferation inhibitor is provided.
9. The use of retinoic acid in the preparation of a medicine for regenerating teeth as claimed in claim 6, wherein the concentration of retinoic acid promoting pharyngeal tooth mineralization is ≥ 3 × 10-7M。
10. The use of retinoic acid as claimed in claim 7 or 8 for preparing a medicine for regenerating teeth, wherein the concentration of the retinoic acid inhibitor for inhibiting migration and proliferation of cranial nerve spine cells is 0.5 × 10-7M-1×10- 7M。
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