阅读说明：本技术 一种支架材料的制备及其应用 (Preparation and application of stent material ) 是由 蒋欣泉 文晋 吴千驹 王笑 于 2020-04-07 设计创作，主要内容包括：本发明提供一种支架材料的制备方法,包括以下步骤：1)将锶化物、P123、正硅酸乙酯、硝酸钙、磷酸三乙酯、盐酸在乙醇中溶解后进行第一次煅烧,获得生物玻璃粉末；2)将生物玻璃粉末与聚乙烯吡咯烷酮、聚乙二醇混合后注入模具中压制成支架前体,再进行第二次煅烧,即得支架材料。本发明进一步提供了一种支架材料及其应用。本发明提供的一种支架材料的制备方法,将具备生物学效应的Sr离子合成至骨组织工程支架中,显著提升骨组织再生效果,可针对于骨质疏松情况下的骨缺损进行再生修复。(The invention provides a preparation method of a bracket material, which comprises the following steps: 1) dissolving strontium compound, P123, ethyl orthosilicate, calcium nitrate, triethyl phosphate and hydrochloric acid in ethanol, and then calcining for the first time to obtain bioglass powder; 2) and mixing the bioglass powder with polyvinylpyrrolidone and polyethylene glycol, injecting the mixture into a mold, pressing the mixture into a support precursor, and calcining the support precursor for the second time to obtain the support material. The invention further provides a stent material and application thereof. According to the preparation method of the scaffold material provided by the invention, Sr ions with biological effects are synthesized into the bone tissue engineering scaffold, so that the bone tissue regeneration effect is obviously improved, and the bone defect under the condition of osteoporosis can be subjected to regeneration repair.)

1. A preparation method of a stent material comprises the following steps:

1) dissolving strontium compound, P123, ethyl orthosilicate, calcium nitrate, triethyl phosphate and hydrochloric acid in ethanol, and then calcining for the first time to obtain bioglass powder;

2) mixing the bioglass powder obtained in the step 1) with polyvinylpyrrolidone and polyethylene glycol, injecting the mixture into a mold to be pressed into a stent precursor, and then carrying out secondary calcination to obtain the stent material.

2. The method for preparing a scaffold material according to claim 1, wherein step 1) comprises any one or more of the following conditions:

A1) the strontium compound is SrCl₂.6H₂O；

A2) The P123 is a triblock copolymer of polyethylene oxide-polypropylene oxide-polyethylene oxide;

A3) the calcium nitrate is Ca (NO)₃)₂·4H₂O；

A4) The hydrochloric acid is 0.5-1.5mol/L HCl aqueous solution;

A5) the weight ratio of the strontium compound to the P123 to the ethyl orthosilicate to the calcium nitrate to the triethyl phosphate to the hydrochloric acid to the ethanol is 1.3-1.4:7-9:13-14:2.5-3.0:1.4-1.5:1.5-2.5: 120;

A6) the temperature of the first calcination is 650-750 ℃;

A7) the time of the first calcination is 4-6 hours.

3. The method for preparing a scaffold material according to claim 1, wherein step 2) comprises any one or more of the following conditions:

B1) the weight ratio of the bioglass powder to the polyvinylpyrrolidone and the polyethylene glycol is 1:0.1-0.3: 0.5-1.5;

B2) the pressing pressure is 4-6 MPa;

B3) the temperature of the second calcination is 580-620 ℃;

B4) the time of the second calcination is 4-6 hours.

4. A scaffold material prepared according to the method of any one of claims 1-3.

5. Use of a scaffold material according to claim 4 as a bone scaffold in osteoporotic bone tissue.

6. Use according to claim 5, wherein the osteoporotic bone tissue is bone tissue of the oromaxillofacial portion of a human body having osteoporosis.

7. A critical defect model of osteoporosis comprising the scaffold material of claim 4.

8. The method for constructing a critical defect model for osteoporosis as claimed in claim 7, wherein the bone defect is prepared on the bone tissue of a laboratory mouse, and the scaffold material as claimed in claim 4 is used for filling and repairing.

9. Use of a critical defect model for osteoporosis according to claim 7 to examine the effect of bone defect repair in the case of osteoporosis.

Technical Field

The invention belongs to the technical field of medical materials, relates to preparation and application of a scaffold material, and particularly relates to a preparation method of the scaffold material and application of the scaffold material in osteoporotic bone tissues.

Background

The oral and maxillofacial region is an important region of the human body, in which the deep jaw system forms the basis of the maxillofacial region and plays an important physiological role. However, trauma, infection, tumor, etc., all cause defects in the bone tissue. At present, osteoporosis is a disease widely existing in the middle-aged and elderly population, and studies have shown that the strength of bones in the body is reduced in this state, and the regeneration ability of the damaged bone tissue is significantly reduced. Therefore, bone defect repair for patients with osteoporosis becomes a major problem.

At present, the defects of the bone tissues of the maxillofacial region are mainly repaired by using a tissue engineering scaffold material, and a certain effect is achieved. Among them, Mesoporous Bioactive Glass scaffold (MBG) is well-developed. However, in the case of metabolic bone diseases (e.g., osteoporosis) caused by some specific diseases, MBG has insufficient osteoinduction and is difficult to support a good bone repair effect. Therefore, as research progresses, the use of various growth factors, such as Bone Morphogenetic Protein (BMP), etc., which are beneficial to bone tissue regeneration, in combination with scaffold materials has been in moderate use. However, the preparation method is complicated and high in cost, and the problems of insufficient concentration, inactivation and the like of possible local growth factors exist in the application process, so that the preparation method is limited to be widely applied clinically.

Disclosure of Invention

In view of the above-mentioned shortcomings of the prior art, the present invention aims to overcome the disadvantages of relatively complicated scaffold modified by growth factors, and the like, and provides a preparation method of a scaffold material, which utilizes a simple physical and chemical method to carry out ion modification on the scaffold material, and is used as a bone scaffold material for repairing bone defects in osteoporosis. Meanwhile, the invention also creates an osteoporosis bone tissue defect model containing the bracket material, and the osteoporosis bone tissue defect model is used for evaluating the bone defect repairing effect.

In order to achieve the above and other related objects, a first aspect of the present invention provides a method for preparing a stent material, comprising the steps of:

1) dissolving strontium compound, P123, ethyl orthosilicate, calcium nitrate, triethyl phosphate and hydrochloric acid in ethanol, and then calcining for the first time to obtain bioglass powder;

2) mixing the bioglass powder obtained in the step 1) with polyvinylpyrrolidone and polyethylene glycol, injecting the mixture into a mold to be pressed into a stent precursor, and then carrying out secondary calcination to obtain the stent material.

Preferably, in step 1), the strontium compound is SrCl₂.6H₂O。

Strontium ions (Sr) in the strontium compound are a trace element existing in a body and have an important effect on bone tissue metabolism. Sr can promote the proliferation of osteoblast and has obvious inhibiting effect on osteoclast, and Sr is one of the important components in strontium ranelate as one of the medicine for osteoporosis.

Preferably, in the step 1), the P123 is a polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer, the molecular formula of the P123 is PEO-PPO-PEO, the CAS number of the P123 is 106392-12-5, and the P123 can be used as a pore forming agent for manufacturing a loose porous structure.

More preferably, the number average molecular weight of the polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer is 5600-6000, and the contents of polyethylene oxide, polypropylene oxide and polyethylene oxide in the polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer respectively account for 30-31%, 39-40% and 30-31%.

Preferably, in step 1), the calcium nitrate is Ca (NO)₃)₂·4H₂O。

Preferably, in step 1), the hydrochloric acid is 0.5-1.5M (mol/L) hydrochloric acid aqueous solution. The solute HCl in the hydrochloric acid aqueous solution is commercial concentrated hydrochloric acid.

Preferably, in the step 1), the weight ratio of the strontium compound, the P123, the ethyl orthosilicate, the calcium nitrate, the triethyl phosphate, the hydrochloric acid and the ethanol is 1.3-1.4:7-9:13-14:2.5-3.0:1.4-1.5:1.5-2.5: 120.

More preferably, the weight ratio of the strontium compound, the P123, the ethyl orthosilicate, the calcium nitrate, the triethyl phosphate, the hydrochloric acid and the ethanol is 1.34:8:13.4:2.8:1.46:2: 120.

Preferably, in step 1), the dissolving is performed at room temperature with sufficient stirring. The room temperature is 20-30 ℃.

Preferably, in the step 1), the temperature of the first calcination is 650-750 ℃. Preferably, in step 1), the time of the first calcination is 4 to 6 hours, preferably 5 hours.

Preferably, in the step 2), the weight ratio of the bioglass powder to the polyvinylpyrrolidone and the polyethylene glycol is 1:0.1-0.3: 0.5-1.5. More preferably, the weight ratio of the bioglass powder to the polyvinylpyrrolidone and the polyethylene glycol is 1:0.2: 1.

Preferably, in the step 2), the number average molecular weight of the polyvinylpyrrolidone is 53000-55000. The CAS number of the polyvinylpyrrolidone is 9003-39-8.

Preferably, in step 2), the number average molecular weight of the polyethylene glycol is 5800-6200. The CAS number of the polyethylene glycol is 25322-68-3.

Preferably, in step 2), the mold is in the shape of a cylinder, and the diameter of the cylinder is 4-6 mm.

Preferably, in step 2), the pressure of the pressing is 4 to 6MPa, preferably 5 MPa.

Preferably, in step 2), the temperature of the second calcination is 580-620 ℃.

Preferably, in step 2), the time of the second calcination is 4 to 6 hours, preferably 5 hours.

The second aspect of the invention provides a stent material prepared by the preparation method.

In a third aspect the present invention provides the use of a scaffold material as a bone scaffold in osteoporotic bone tissue.

The osteoporotic bone tissue is a whole body bone tissue of a human body having osteoporosis. Osteoporosis can affect bone tissue throughout the body.

Preferably, the osteoporotic bone tissue is bone tissue of an oral maxillofacial portion of a human body having osteoporosis. Osteoporosis will result in a reduction in the ability of the oral maxillofacial bone to repair.

The invention provides a osteoporosis critical defect model, which comprises the support material.

The fifth aspect of the invention provides a method for constructing a osteoporosis critical defect model, wherein bone defects are manufactured on bone tissues of a laboratory mouse, and the support material is adopted for filling and repairing.

The bone tissue of the experimental mouse is exposed after the skin and fascia of the experimental mouse are cut after the experimental mouse is anesthetized. The skin of the experimental mouse needs to be sutured after the scaffold material is filled and repaired. The incision part of the experimental mouse is the median of the brain, and the incision of the experimental mouse is blunt separation.

Preferably, the experimental mice are female SD rats 12 weeks after bilateral ovariectomy.

Preferably, the scaffold material to be filled and repaired has a diameter of 4.5-5.5mm, preferably 5 mm.

The sixth aspect of the invention provides the use of the osteoporosis critical defect model described above to examine the effect of bone defect repair in the case of osteoporosis.

As described above, according to the preparation method of the bone scaffold material provided by the invention, the biological effect of Sr ions is fully utilized, Sr ions with biological effect are synthesized into the bone tissue engineering scaffold, and the Sr ion-modified bioactive glass scaffold material is prepared, so that the osteogenic activity of the material can be improved, the bone tissue regeneration effect can be remarkably improved, the bone defect under the condition of osteoporosis can be regenerated and repaired, and the clinical application of the material is promoted.

Drawings

Fig. 1a and 1b show osteoporosis critical defect model diagrams, wherein fig. 1a is a schematic diagram of a skull defect model of a mouse filled and repaired by using a Sr ion-loaded scaffold material prepared by the invention, and fig. 1b is a schematic diagram of a skull defect model of a mouse filled and repaired by using a pure MBG scaffold material and a Sr ion-loaded scaffold material prepared by the invention from left to right.

Fig. 2a and 2b are graphs showing the effect comparison after the osteoporosis critical defect model repair, wherein fig. 2a is a graph showing the repair effect of a pure MBG stent material, and fig. 2b is a graph showing the repair effect of a Sr ion-loaded stent material prepared according to the present invention.

Fig. 3a and 3b are graphs showing The comparison of The results of quantitative analysis of new bone tissue using CT measurement, in which fig. 3a is a graph comparing The relative bone volume of bone volume to total volume (BV/TV) measured by CT of simple MBG scaffold material and Sr ion-loaded scaffold material, and fig. 3b is a graph comparing Trabecular thickness (tb.th) measured by CT of simple MBG scaffold material and Sr ion-loaded scaffold material.

Fig. 4a, 4b, 4c and 4d are views showing the detection of new bone tissue by HE staining, in which fig. 4a is a view of HE staining for bone defect repair using a simple MBG scaffold, fig. 4b is a view of HE staining for bone defect repair using a Sr ion-modified scaffold, fig. 4c is an enlarged view of a target area of a black frame in fig. 4a, and fig. 4d is an enlarged view of a target area of a black frame in fig. 4 b.

Detailed Description

The present invention is further illustrated below with reference to specific examples, which are intended to be illustrative only and not to limit the scope of the invention.

The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention.

It should be understood that the processing equipment or devices not specifically mentioned in the following examples are conventional in the art; all pressure values and ranges refer to relative pressures.

Furthermore, it is to be understood that one or more method steps mentioned in the present invention does not exclude that other method steps may also be present before or after the combined steps or that other method steps may also be inserted between these explicitly mentioned steps, unless otherwise indicated; it is also to be understood that a combined connection between one or more devices/apparatus as referred to in the present application does not exclude that further devices/apparatus may be present before or after the combined device/apparatus or that further devices/apparatus may be interposed between two devices/apparatus explicitly referred to, unless otherwise indicated. Moreover, unless otherwise indicated, the numbering of the various method steps is merely a convenient tool for identifying the various method steps, and is not intended to limit the order in which the method steps are arranged or the scope of the invention in which the invention may be practiced, and changes or modifications in the relative relationship may be made without substantially changing the technical content.

Example 1

1.34g SrCl at room temperature₂.6H₂O, 8g P123, 13.4g tetraethoxysilane, 2.8gCa (NO)₃)₂·4H₂O, 1.46g of triethyl phosphate and 2g of 1mol/L hydrochloric acid were dissolved in 120g of ethanol with stirring. The resulting solution was calcined at 700 ℃ for 5 hours to obtain a bioglass powder sample # 1. Wherein, the P123 is polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer with the number average molecular weight of 5800, and the contents of polyethylene oxide, polypropylene oxide and polyethylene oxide in the polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer respectively account for 30.1%, 39.8% and 30.1%.

Mixing the biological glass powder sample No. 1 with polyvinylpyrrolidone and polyethylene glycol at a weight ratio of 1:0.2:1, injecting into a cylindrical mold with a diameter of 5mm, pressing under a pressure of 5MPa to prepare a stent precursor, and calcining at 600 ℃ for 5 hours to obtain a stent material sample No. 1. Wherein, the number average molecular weight of the polyvinylpyrrolidone is 54000, and the number average molecular weight of the polyethylene glycol is 6000.

Example 2

1.36g SrCl at room temperature₂.6H₂O, 8.2g P123, 13.6g tetraethoxysilane, 2.7gCa (NO)₃)₂·4H₂O, 1.44g of triethyl phosphate and 2.1g of 1mol/L hydrochloric acid were dissolved in 120g of ethanol with stirring. Subjecting the obtainedThe solution was calcined at 750 ℃ for 4.5 hours to obtain a bioglass powder sample # 2. Wherein, the P123 is polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer with the number average molecular weight of 5900, and the contents of polyethylene oxide, polypropylene oxide and polyethylene oxide in the polyethylene oxide-polypropylene oxide-polyethylene oxide triblock copolymer respectively account for 30.3%, 39.4% and 30.3%.

Mixing the biological glass powder sample No. 2 with polyvinylpyrrolidone and polyethylene glycol at a weight ratio of 1:0.25:1, injecting into a cylindrical mold with a diameter of 5mm, pressing under a pressure of 5MPa to prepare a stent precursor, and calcining at 580 ℃ for 4.5 hours to obtain a stent material sample No. 2. Wherein, the number average molecular weight of the polyvinylpyrrolidone is 53000, and the number average molecular weight of the polyethylene glycol is 6100.

Example 3

Taking female SD rats 12 weeks after cutting ovaries on both sides, cutting skin and fascia in the middle of the brain along the middle after anesthesia, exposing bone tissues after blunt separation, manufacturing bilateral bone defects, filling a skull defect part of the experimental rat with the scaffold material sample 1# prepared in example 1 as a bone scaffold material for repair, wherein the diameter of the scaffold material sample 1# filled in the bone defect part is 5mm, and performing conventional suture to form an osteoporosis critical defect model 1, wherein the specific situation is shown in figure 1 a.

Comparative example 1

Taking a female SD rat, cutting the median skin and fascia of the brain 12 weeks after cutting ovaries on both sides, anesthetizing, incising the median skin and fascia along the median, carrying out blunt separation, exposing bone tissues, manufacturing bilateral bone defects, filling and repairing the left side defect part of the skull of the experimental rat by using a pure MBG scaffold material as a bone scaffold material, filling and repairing the right side defect part of the skull of the experimental rat by using the scaffold material sample 1# prepared in the embodiment 1 as a bone scaffold material, wherein the diameters of the pure MBG scaffold material filled in the bone defect part and the scaffold material sample 1# are both 5mm, and carrying out conventional suture to form a osteoporosis critical defect model 2, wherein the specific situation is shown in figure 1 b.

Comparative test example 1

The osteoporosis critical defect model 2 obtained in comparative example 1 was examined for bone repair effect at 2 months after the operation. In the osteoporosis state, as shown in fig. 2a, in the left side defect site of the osteoporosis critical defect model 2, the ability of repairing bone defects by using a pure MBG scaffold material is relatively insufficient, and less new bone tissue formation is observed. As shown in fig. 2b, in the right defect site of the osteoporosis critical defect model 2, the Sr ion-modified bioactive scaffold material prepared by the present invention can significantly improve the bone tissue regeneration effect, and the newly-grown bone tissue at the bone defect site is significantly increased. In fig. 2a, 2b, the arrows represent new bone tissue.

CT is adopted to respectively determine The osteoporosis critical defect model 2 of 2 months after operation, The CT result is subjected to quantitative analysis, and as can be seen from figure 3a, in The osteoporosis critical defect model 2, The relative bone volume of bone volume to total volume (BV/TV) of The right side defect part obviously exceeds The left side defect part; as can be seen from fig. 3b, in the osteoporosis critical defect model 2, Trabecular thickness (tb.th) of the right side defect portion also exceeded that of the left side defect portion. Therefore, the Sr ion modified bioactive scaffold material has a remarkable effect of promoting bone tissue regeneration. In the above fig. 3a, 3b, p <0.05 indicates that the two groups of material new bone tissue have statistical significance in relative bone volume and trabecular thickness.

The HE staining was used to detect the condition of the new bone tissue in the osteoporosis critical defect model 2 at 2 months after the operation, and the detection results are shown in FIGS. 4a, 4b, 4c, and 4 d. In fig. 4a, 4b, 4c, 4d, the arrows represent new bone tissue. Fig. 4a and 4c show that in the osteoporosis critical defect model 2, the left defect site was repaired using a simple MBG scaffold, and fig. 4b and 4d show that the right defect site was repaired using a Sr ion-modified bioactive scaffold. The comparison of the two shows that the bioactive scaffold material modified by Sr ions in the osteoporosis critical defect model 2 is adopted to repair the right side defect part, the area of the new bone tissue is obviously increased, and the repair effect is superior to that of a pure MBG scaffold material for repairing the left side defect part.

In conclusion, the preparation method of the scaffold material provided by the invention synthesizes Sr ions with biological effects into the bone tissue engineering scaffold, obviously improves the bone tissue regeneration effect, and can be used for performing regeneration repair on bone defects under the condition of osteoporosis. Therefore, the invention effectively overcomes various defects in the prior art and has high industrial utilization value.

The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.