CN103767881A - Dental pulp capping preparation and preparation method thereof - Google Patents
Dental pulp capping preparation and preparation method thereof Download PDFInfo
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- CN103767881A CN103767881A CN201410004894.1A CN201410004894A CN103767881A CN 103767881 A CN103767881 A CN 103767881A CN 201410004894 A CN201410004894 A CN 201410004894A CN 103767881 A CN103767881 A CN 103767881A
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Abstract
The invention relates to the field of research and development of tissue regeneration materials, and discloses a dental pulp capping preparation and a preparation method thereof. The dental pulp capping preparation is an rhBMP2-PLGA (recombinant human bone morphogenetic protein 2-poly(lactic acid-co-glycolic acid)) sustained-release microsphere. The preparation method comprises the following steps: adding an rhBMP2 solution into a PLGA dichloromethane solution; performing ultrasonic emulsification under an ice bath condition to form uniform primary emulsion, and stirring by using a high-speed homogenizer at 10,000r/min; extracting the stirred primary emulsion and injecting into a PVA (polyvinyl acetate) solution at a uniform speed, and uniformly dispersing to form secondary emulsion; magnetically stirring, centrifuging after the dichloromethane is volatized completely, and collecting precipitate; repeatedly washing with deionized water, lyophilizing to obtain the rhBMP-PLGA sustained-release microsphere, and storing at 4 DEG C. The rhBMP2-PLGA sustained-release microsphere has the sustained-release and long-acting effect, and can be used for protecting the activity of related factors of a genetic recombinant human bone morphogenetic protein 2 to form new dentin, so that the aim of protecting vital pulp can be achieved.
Description
Technical field
The present invention relates to and tissue regeneration material development field, particularly a kind of tooth pulp-cap and preparation method thereof.
Background technology
In disease of pulp of tooth treatment, vital pulp therapy is the method that meets viewpoint biology most, and in its treatment, the effect of pulp-cap is most important.In numerous lid marrow materials at present used, bone morphogenetic protein (Bone Morphogenetic Protein, BMP) is one of material of common concern always.There is report to think that BMP is used as and cover marrow treatment, in short-term, just can form a large amount of reparative dentin sealing dew pith cavities, be conducive to the recovery of dental pulp.But, in the time that BMP applies separately in vivo, will soon be diluted or be destroyed by protease, short to the action time of pulp cells, can not effectively bring into play its due biological property; And can not provide support for dentin forms, it is very restricted in clinical practice.
Summary of the invention
The object of the present invention is to provide a kind of tooth pulp-cap and preparation method thereof, using rhBMP2-PLGA sustained-release micro-spheres material as tooth pulp-cap, can solve the deficiency of BMP in the time of application, can play slow releasing function; Again can protecting group because of recombination human bone shaping protein 2(recombinant human bone morphogenetic protein-2 (rhBMP-2)) the activity of correlation factor, reach the object of protection vital pulp.
In order to achieve the above object, the present invention is achieved by the following technical solutions.
A kind of tooth pulp-cap, is characterized in that, described tooth pulp-cap is rhBMP2-PLGA sustained-release micro-spheres.
The preparation method of above-mentioned tooth pulp-cap, it is characterized in that, comprise the following steps: first rhBMP2 solution is joined in PLGA dichloromethane solution, and under condition of ice bath, ultrasonic emulsification forms uniform colostrum, and under 10000r/min condition, stir with high speed homogenizer; The colostrum extracting after stirring is at the uniform velocity noted in PVA solution, dispersed formation emulsion; Magnetic agitation, makes dichloromethane volatilization completely rear centrifugal, collecting precipitation thing; Deionized water cyclic washing postlyophilization, obtains the rhBMP2-PLGA sustained-release micro-spheres as tooth pulp-cap, 4 ℃ of preservations.
In technique scheme, preferably, quality-the volumetric concentration of rhBMP2 solution is 0.1%(mg/ml), quality-the volumetric concentration of PLGA dichloromethane solution is 2.5%(mg/ml), quality-the volumetric concentration of PVA solution is 1%(mg/ml), wherein the volume ratio of rhBMP2 solution, PLGA dichloromethane solution, PVA solution is 1:10:100.
The present invention is using rhBMP2-PLGA sustained-release micro-spheres as tooth pulp-cap, gene recombinaton Human Bone Morphogenetic Proteins-4 2(recombinant human bone morphogenetic protein-2 (rhBMP-2)) be main body, with Poly(D,L-lactide-co-glycolide (poly (lactic-co-glycolic acid), PLGA) be support, solve the technical problem that rhBMP-2 destroys by rapid dilution with by protease in vivo, there is slow release long-acting effect, protecting group is because of recombination human bone shaping protein 2(recombinant human bone morphogenetic protein-2 (rhBMP-2)) the activity of correlation factor, form new dentin, to reach the object of protection vital pulp.
In addition better, biodegradable, the avirulence of PLGA biocompatibility; And itself and catabolite thereof all to polypeptide class, albumen etc. without obvious untoward reaction, safe and reliable, and the approval that obtains U.S. FDA is for clinical.
Accompanying drawing explanation
Below in conjunction with the drawings and specific embodiments, the present invention is described in further details.
Fig. 1 is rhBMP2-PLGA sustained-release micro-spheres stereoscan photograph figure.
Fig. 2 is rhBMP2-PLGA sustained-release micro-spheres particle size distribution figure.
Fig. 3 is the release in vitro broken line graph of rhBMP2-PLGA sustained-release micro-spheres.
Fig. 4 is blank group experimental result picture, in figure, forms without sclerous tissues.
Fig. 5 is rhBMP2 group experimental result picture, has dentin calcification bridge in figure, but relatively less.
Fig. 6 is rhBMP2-PLGA sustained-release micro-spheres group experimental result picture, has dentin to generate in figure, forms complete calcification bridge.
Fig. 7 is collagen group experimental result picture, can observe complete dentine bridge and form in figure;
Fig. 8 is antibiotic group experimental result picture, has the sclerous tissues of calcification to generate in figure, but without complete dentin calcification bridge.
Fig. 9 is antibiotic and rhBMP2-PLGA sustained-release micro-spheres mixing group experimental result picture, can observe complete dentin calcification bridge in figure.
Figure 10 is collagen and rhBMP2-PLGA sustained-release micro-spheres mixing group experimental result picture, can observe complete dentin calcification bridge and generate, and pulp cavity is interior without calcification in figure.
Figure 11 is the block diagram that each treated animal is tested dentine bridge's thickness.
The specific embodiment
In the present invention, PLGA, PVA buy the company in SIGMA; RhBMP2 is purchased from Shanghai Ruibang Biological Material Co., Ltd.; RhBMP2ELISA detection kit is purchased from Wuhan doctor's moral company.Adopt (oil-in-water, Water-In-Oil) W/O/W emulsion solvent evaporation method to prepare rhBMP2-PLGA sustained-release micro-spheres.
1, experimental technique
(1) experiment preparation:
By 200ul containing 0.1%(mg/ml) rhBMP2 solution (the interior water of quality-volumetric concentration, be made as W1) join 2ml and contain 2.5%(mg/ml) the PLGA dichloromethane solution (oil phase of quality-volumetric concentration, be made as O) in, under condition of ice bath, ultrasonic emulsification forms uniform colostrum (W1/O), stirs immediately with high speed homogenizer under 10000r/min condition; With syringe extract colostrum after stirring slowly at the uniform velocity note in to 20ml containing 1%(mg/ml) in the PVA solution (outer water, is made as W2) of quality-volumetric concentration, dispersed 2min forms the emulsion of W1/O/W2; Magnetic agitation 3～4h, makes after dichloromethane volatilization completely the centrifugal 5min of 15000r/min, collecting precipitation thing; Deionized water is washed 5 times repeatedly, and lyophilization 48h obtains rhBMP2-PLGA sustained-release micro-spheres, 4 ℃ of preservations.
(2) morphologic observation:
Get appropriate rhBMP2-PLGA sustained-release micro-spheres and be dispersed on the metallic plate that posts double faced adhesive tape, after metal spraying, observe its configuration of surface with surface sweeping Electronic Speculum (SEM).
(3) microspherulite diameter and measure of spread:
Get appropriate rhBMP2-PLGA sustained-release micro-spheres and be scattered in ultra-pure water, after sonic oscillation is uniformly dispersed, in laser particle size analyzer (Ma Erwen Nano ZS-90), measure its distribution and particle diameter.
(4) measure envelop rate and drug loading:
Precision takes rhBMP2-PLGA sustained-release micro-spheres 20mg, adds in 2mL0.1mol/L NaOH (containing 0.5%SDS) solution, in 37 ℃ of water bath with thermostatic control oscillation incubation 24h, microsphere is dissolved completely; After being neutralized to pH value and being 7.0 with 0.1mol/LHCl, the centrifugal 5min of 12000r/min, gets supernatant, measures the content of rhBMP2 in medicine carrying microballoons with ELISA, and concrete operations are carried out according to the explanation on test kit.And respectively by following formula computational envelope rate and drug loading:
Microsphere envelop rate (%)=(content of dispersion/dosage in microsphere) × 100%
Microsphere drug loading (%)=(content of dispersion/microspheres quality in microsphere) × 100%
(5) calculate burst size:
Accurately measuring the PBS buffer (20mmol) that rhBMP2-PLGA sustained-release micro-spheres 10mg is dissolved in 5ml pH7.4 is release medium, and vibration at 37 ℃ is taken out respectively supernatant 200ul respectively at 1,3,7,14, when 28d and supplemented the release medium of fresh equivalent.Supernatant is measured the concentration of rhBMP2 by rhBMP2ELISA, and calculates burst size.
(6) zoopery:
Select 2 healthy adult beasle dogs (being purchased from Xi'an Di Lepu living resources development corporation, Ltd.), SPF level (safety and Health rank), body weight is respectively 10.8kg, 11.2kg, age 12-14 month, be male, the complete and Ya Ti periodontal tissue health of denture, whole body is without disease, experiment prospective adaptation is raised 1～2 week, after disposition is stable for experiment.
Every dog is chosen upper and lower both sides premolar teeth, front tooth, totally 42 teeth, 42 teeth are divided into 7 groups at random by table of random number method: rhBMP2 group, rhBMP2-PLGA sustained-release micro-spheres, collagen, dentistry antibiotic, dentistry antibiotic ﹢ rhBMP2-PLGA sustained-release micro-spheres, collagen ﹢ rhBMP2-PLGA sustained-release micro-spheres, blank group; Every group of 6 teeth.After experiment, all raise after 3 months and put to death animal.
Before the operation of lid marrow, 12h starts fasting.Within preoperative 30 minutes, sleep new injection by the capable preoperative intramuscular anesthesia of the dosage of 0.1ml/Kg by speed, in art, suitably appending plastic injection quality-volumetric concentration according to anesthesia situation is 3% pentobarbital sodium, to maintain anaesthetic effect.After anesthesia, experimental dog being lain on the back and is fixed on operating-table, is that 3% hydrogen peroxide liquid rinses oral cavity by molar concentration, and is that the ethanol that 1% iodine tincture and volumetric concentration are 75% carries out disinfection to oral cavity and experiment tooth by quality-volumetric concentration.Occlusal surface at experiment tooth bores and reveals marrow diameter 1mm with high speed turbine mobile phone bead, softly rinse with the sodium hypochlorite that 10ml quality-volumetric concentration is 2.5%, then with 2ml quality-volumetric concentration be 17%EDTA(Chinese:
ethylenediaminetetraacetic acid) rinse, hemostasis after flushing, dry, is placed in pulp-cap to reveal marrow place, puts into big or small essentially identical gelfoam, glass ionomer.Concrete operation step is as follows:
Blank group: directly place gelfoam, glass ionomer, and be coated with vaseline every wet.
RhBMP2 group: after being dipped in to gelfoam, rhBMP2 is directly placed in dental pulp section, glass ionomer, and be coated with vaseline every wet.
RhBMP2-PLGA sustained-release micro-spheres group: rhBMP2-PLGA sustained-release micro-spheres is directly placed in to dental pulp section, places gelfoam, glass ionomer, and be coated with vaseline every wet.
Collagen group: dip appropriate collagen with gelfoam and be placed in dental pulp section, glass ionomer, and be coated with vaseline every wet.
Antibiotic group: by two kinds of preparation mix homogeneously, and leach and be placed in right amount on dental pulp section with gelfoam, glass ionomer, and be coated with vaseline every wet.
Antibiotic and rhBMP2-PLGA sustained-release micro-spheres mixing group: will be soaked with appropriate antibiotic gelfoam and dip the white powder (after lyophilization) of appropriate rhBMP2-PLGA sustained-release micro-spheres, be placed on dental pulp section, glass ionomer, and be coated with vaseline every wet.
Collagen and rhBMP2-PLGA sustained-release micro-spheres mixing group: the gelfoam that is soaked with appropriate collagen is dipped to appropriate rhBMP2-PLGA sustained-release micro-spheres white powder, be placed on dental pulp section, glass ionomer, and be coated with vaseline every wet.
In this experimental example, antibiotic is metronidazole, ciprofloxacin, minocycline, minocycline etc. mass parts mixture; And whole operation process completes by same people, within postoperative first three day, animal enters soft diet, injection of antibiotic three days.The situation of charges and whole body mental status in the oral cavity of postoperative routine observation experimental dog.Postoperative to experiment tooth shooting X-ray film.After postoperative three months, sacrifice of animal is drawn materials, with MicroCT scanning, sclerous tissues forms.
2, experimental result
(1) after the rhBMP2-PLGA sustained-release micro-spheres lyophilizing of preparing under Optimization Technology, be white in color Powdered, without collapse phenomenon; RhBMP2-PLGA sustained-release micro-spheres form rounding is even, smooth surface, without adhesion as Fig. 1.
(2) rhBMP2-PLGA sustained-release micro-spheres particle size distribution is normal distribution, and as shown in Figure 2, its distribution is concentrated, and most of microspherulite diameter is distributed between 3um～4um.
(3) envelop rate 72.5% ± 4.17% of rhBMP2-PLGA sustained-release micro-spheres, the carrying drug ratio of microsphere is 0.38% ± 0.24%.The sustained-release micro-spheres burst size that is loaded with rhBMP2 when release in vitro 1d is 19.36% ± 1.83%, show as the prominent property released and discharge, this is mainly that release ratio is slower subsequently due to due to the drug release on sustained-release micro-spheres surface, but Cumulative release amount continues to increase, during to 28d, reach 52.76% ± 3.7%.As shown in Figure 3.
(4) results of animal: along experiment tooth words to tooth long axile, MicroCT observes and measure newborn Dentinal thickness, and obtains the meansigma methods of each group.
Blank group: as shown in Figure 4, do not observe sclerous tissues and form;
RhBMP2 group: as shown in Figure 5, can observe complete dentin calcification bridge, but relatively less.
RhBMP2-PLGA sustained-release micro-spheres group: as shown in Figure 6, can observe complete calcification bridge and generate, pulp cavity is without calcification;
Collagen group: as shown in Figure 7, can observe complete dentine bridge and form;
Antibiotic group: as shown in Figure 8, the sclerous tissues that can observe calcification generates, but without complete dentin calcification bridge;
Antibiotic and rhBMP2-PLGA sustained-release micro-spheres mixing group: as shown in Figure 9, can observe complete dentin calcification bridge and generate, and pulp cavity is interior without calcification;
Collagen and rhBMP2-PLGA sustained-release micro-spheres mixing group: as shown in figure 10, can observe complete dentin calcification bridge and generate, and pulp cavity is interior without calcification.
As shown in figure 11, rhBMP2 group is mixed between group, antibiotic and rhBMP2-PLGA sustained-release micro-spheres mixing group and is had and have significant difference (P<0.05) with collagen group, rhBMP2-PLGA sustained-release micro-spheres group, antibiotic and rhBMP2-PLGA sustained-release micro-spheres.
3, experiment conclusion
This experiment is by being used the experimental result of DNAcarrier free rhBMP2 and other groups to compare rear discovery, the simple rhBMP2 that uses also has certain effect to the formation of induction reparative dentin, but its effect is far away not as good as other groups that used rhBMP2-PLGA sustained-release micro-spheres, and concrete reason is that the Organic substance in pulp tissue destroys and causes to some extent the activity of rhBMP2.
This experimental result shows that rhBMP2-PLGArhBMP2-PLGA sustained-release micro-spheres and composite dental antibiotic carry out organizing strong compared with other to the induction of differentiation of pulp cells after pulp capping, the dentine bridge forming in same time is thicker and complete, there is good lid marrow effect, for it is for the clinical feasibility that provides.
Those skilled in the art can use for reference content herein, suitably improve technological parameter and realize, or do not depart from content of the present invention, spirit and scope and methods and applications as herein described are changed or suitably changed and combination, realize and apply the technology of the present invention.Special needs to be pointed out is, above-mentioned all similar replacements and change apparent to those skilled in the artly, they are all deemed to be included in protection domain of the present invention.
Claims (3)
1. a tooth pulp-cap, is characterized in that, described tooth pulp-cap is rhBMP2-PLGA sustained-release micro-spheres.
2. the preparation method of tooth pulp-cap as claimed in claim 1, it is characterized in that, comprise the following steps: first rhBMP2 solution is joined in PLGA dichloromethane solution, and under condition of ice bath, ultrasonic emulsification forms uniform colostrum, and under 10000r/min condition, stir with high speed homogenizer; The colostrum extracting after stirring is at the uniform velocity noted in PVA solution, dispersed formation emulsion; Magnetic agitation, makes dichloromethane volatilization completely rear centrifugal, collecting precipitation thing; Deionized water cyclic washing postlyophilization, obtains the rhBMP2-PLGA sustained-release micro-spheres as tooth pulp-cap, 4 ℃ of preservations.
3. the preparation method of tooth pulp-cap as claimed in claim 2, it is characterized in that, quality-the volumetric concentration of rhBMP2 solution is 0.1%(mg/ml), quality-the volumetric concentration of PLGA dichloromethane solution is 2.5%(mg/ml), quality-the volumetric concentration of PVA solution is 1%(mg/ml), wherein the volume ratio of rhBMP2 solution, PLGA dichloromethane solution, PVA solution is 1:10:100.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104645419A (en) * | 2014-12-02 | 2015-05-27 | 中国人民解放军第四军医大学 | Preparation method of porous titanium-alloy femoral head support rod in bionic bone trabecula structure |
| CN112569122A (en) * | 2020-12-10 | 2021-03-30 | 杭州彗搏科技有限公司 | Notoginsenoside-hydrogel based marrow capping agent material and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1652829A (en) * | 2002-05-13 | 2005-08-10 | 东芝陶瓷株式会社 | Member for regenerating joint cartilage and process for producing the same, method of regenerating joint cartilage and artificial cartilage for transplantation |
| CN102755669A (en) * | 2012-07-16 | 2012-10-31 | 姚琦 | Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere |
| CN103027740A (en) * | 2013-01-17 | 2013-04-10 | 北京中奥汇成生物材料科技有限公司 | Absorbable bone screw with slow-release biological factor and preparation method thereof |
-
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- 2014-01-06 CN CN201410004894.1A patent/CN103767881A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1652829A (en) * | 2002-05-13 | 2005-08-10 | 东芝陶瓷株式会社 | Member for regenerating joint cartilage and process for producing the same, method of regenerating joint cartilage and artificial cartilage for transplantation |
| CN102755669A (en) * | 2012-07-16 | 2012-10-31 | 姚琦 | Preparation method and application of fibrin glue composite recombinant human bone morphogenetic protein-2 (rhBMP-2) microsphere |
| CN103027740A (en) * | 2013-01-17 | 2013-04-10 | 北京中奥汇成生物材料科技有限公司 | Absorbable bone screw with slow-release biological factor and preparation method thereof |
Non-Patent Citations (1)
| Title |
|---|
| 柴雪: "rhBMP2 缓释微球的制备及其用于盖髓剂的实验研究", 《万方数据》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104645419A (en) * | 2014-12-02 | 2015-05-27 | 中国人民解放军第四军医大学 | Preparation method of porous titanium-alloy femoral head support rod in bionic bone trabecula structure |
| CN112569122A (en) * | 2020-12-10 | 2021-03-30 | 杭州彗搏科技有限公司 | Notoginsenoside-hydrogel based marrow capping agent material and preparation method thereof |
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Application publication date: 20140507 |