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CN-121971684-A - Absorbable bone wax for promoting bone healing and preparation method thereof

CN121971684ACN 121971684 ACN121971684 ACN 121971684ACN-121971684-A

Abstract

The invention relates to absorbable bone wax for promoting bone healing and a preparation method thereof, belonging to the technical field of bone healing materials, the bone wax comprises degradable copolymer, PTMC, magnesium-calcium silicate-biological peptide composite microsphere, mesoporous silica-supported Notoginseng radix total saponin and hydrogenated castor oil. The degradable copolymer is prepared by reacting PEG, stannous octoate, D, L-lactide and epsilon-caprolactone and precipitating. The magnesium-calcium silicate-biological peptide composite microsphere is prepared by loading biological peptide on a porous magnesium-calcium silicate microsphere and then coating with polydopamine. The biological peptide is a peptide segment of 1 kDa-5 kDa, which is obtained by step-by-step enzymolysis of urechis unicinctus by papain and trypsin. The components are compounded according to a specific proportion to prepare the bone wax, so that the limitations of poor degradability and single function of the traditional bone wax are broken through, and the integrated functions of hemostasis, degradation and high-efficiency bone induction are realized. By the cooperation of the controllable degradation base material and the high-activity functional component, the accurate matching of the degradation rate and the bone regeneration period is achieved.

Inventors

  • LIAO SHUHUI
  • LIU ZHUOFAN
  • ZHENG LONGPO
  • YANG QINGCHENG
  • QIN KELIN
  • LIAO XIANCHUAN
  • CHEN JING
  • SHU JINRONG
  • QIN YI
  • LIU KE

Assignees

  • 湖北双星药业股份有限公司

Dates

Publication Date
20260505
Application Date
20260407

Claims (10)

  1. 1. The absorbable bone wax for promoting bone healing is characterized by comprising (by mass ratio) a degradable copolymer (25-30), a degradable copolymer (8-10), a degradable copolymer (9-13), a degradable copolymer (3-5), a degradable copolymer (2-4), PTMC (poly (PTMC) - (magnesium-calcium silicate-bio-peptide) composite microspheres, mesoporous silica-supported pseudo-ginseng total saponins and hydrogenated castor oil; the degradable copolymer is prepared by reacting PEG, stannous octoate, D, L-lactide and epsilon-caprolactone according to the mass ratio of 100 (0.3-0.6) (115-135) at 125-135 ℃ and precipitating in normal hexane, wherein the PEG comprises PEG-400 and PEG-1000 with the mass ratio of (2-4) (5-7); The magnesium-calcium silicate-biological peptide composite microsphere is a porous magnesium-calcium silicate microsphere, biological peptide is subjected to vibration adsorption in a phosphate buffer solution according to the mass ratio of (10-15) (0.8-1.2), solid is obtained by centrifugation, dispersed in a Tris-HCl buffer solution with pH of 8.2-8.5 and containing dopamine, and a polydopamine coating layer is prepared by stirring, and the solid is obtained; The biological peptide is a peptide component which is obtained by carrying out enzymolysis on crushed slurry of urechis unicinctus in deionized water for 2 to 2.5 hours at pH of 6.2 to 6.5 and 50 ℃ to 55 ℃ by papain accounting for 1.5 to 2 percent of the mass of the slurry and carrying out enzymolysis on the crushed slurry for 1.5 to 2 hours at pH of 7.8 to 8.2 and 35 ℃ to 40 ℃ by trypsin accounting for 1.8 to 8 ℃ of the mass of the slurry; The porous magnesium-calcium silicate microsphere is prepared by preparing sol according to the mass ratio of (46-52): (7-9): (20-25) of Ca (NO 3 ) 2 ·4H 2 O、Mg(NO 3 ) 2 ·6H 2 O, TEOS), spray drying, calcining at 650-750 ℃ for 3.5-4.5 hours, grinding and sieving with a 150-200 mesh sieve.
  2. 2. The bone healing promoting absorbable bone wax of claim 1, wherein the preparation method of the degradable copolymer comprises the steps of mixing and reacting PEG, stannous octoate, D, L-lactide and epsilon-caprolactone monomers under the protection of argon to obtain a reaction solution, adding anhydrous methylene dichloride, stirring to obtain a diluent, adding the diluent into n-hexane to precipitate under stirring, centrifuging, taking solids, washing with n-hexane, and vacuum drying to obtain the degradable copolymer, wherein the mass ratio of PEG to stannous octoate, L-lactide to epsilon-caprolactone=100 (0.3-0.6) to 115-135) to epsilon-caprolactone.
  3. 3. The bone healing promotion absorbable bone wax according to claim 2, wherein the PEG comprises (2-4): (5-7) of PEG-400 and PEG-1000 by mass ratio, the stirring reaction is carried out at 125-135 ℃ for 22-26 hours, the amount of anhydrous dichloromethane is 8-12 times the mass of the reaction solution, the amount of n-hexane is 8-12 times the volume of the diluent, and the n-hexane is pre-cooled at 2-6 ℃.
  4. 4. The bone healing promoting absorbable bone wax of claim 1, wherein the preparation method of the magnesium-calcium silicate-biological peptide composite microsphere comprises the steps of dispersing the porous magnesium-calcium silicate microsphere in phosphate buffer according to the mass ratio of (10-15) to (120-180) of biological peptide= (0.8-1.2), adding the biological peptide into the phosphate buffer, oscillating for adsorption, centrifuging, taking solids, dispersing in Tris-HCl buffer with pH of 8.2-8.5 and 1mg/mL of dopamine, stirring at room temperature under the dark condition, centrifuging, taking the solids, washing with deionized water, and freeze-drying to obtain the magnesium-calcium silicate-biological peptide composite microsphere.
  5. 5. The bone healing promotion absorbable bone wax according to claim 4, wherein the preparation method of the biological peptide comprises the steps of crushing urechis unicinctus into slurry, adding deionized water, adjusting pH to 6.2-6.5, adding papain with the mass of 1.5-2% of the slurry, performing enzymolysis at 50-55 ℃ for 2-2.5 h, inactivating enzyme, centrifuging at 4000-5000 rpm for 15 min-20 min, taking supernatant, adjusting pH to 7.8-8.2, adding trypsin with the mass of 1-2% of the slurry, performing enzymolysis at 35-40 ℃ for 1.5 h-2 h, inactivating enzyme, centrifuging at 800-8500 rpm for 10 min-15 min, taking supernatant, performing ultrafiltration membrane separation, collecting peptide-segment components with the molecular weight of 1 kDa-5 kDa, and performing freeze drying to obtain the biological peptide.
  6. 6. The bone healing promoting absorbable bone wax of claim 4, wherein the porous magnesium-calcium silicate microsphere is prepared by dissolving Ca (NO 3 ） 2 ·4H 2 O and Mg (NO 3 ） 2 ·6H 2 O) in deionized water to obtain solution A, dissolving TEOS in absolute ethyl alcohol to obtain solution B, adding the solution B into the solution A under stirring to form sol, performing spray drying to obtain precursor powder, heating the precursor powder to 650-750 ℃, calcining for 3.5-4.5 h, cooling, and grinding through a 150-200 mesh sieve to obtain the porous magnesium-calcium silicate microsphere.
  7. 7. The bone healing promoting absorbable bone wax of claim 4, wherein the vibration adsorption is carried out at 35-40 ℃ for 22-26 hours, and the stirring is carried out at 150-200 rpm for 24-28 hours.
  8. 8. The bone healing promoting absorbable bone wax of claim 1, wherein the preparation method of the mesoporous silica-loaded total arasaponin comprises the steps of dissolving CTAB in deionized water, adding aqueous solution of sodium hydroxide, stirring, adding TEOS, stirring for reaction, centrifuging, taking solids, washing and calcining to obtain mesoporous silica nanoparticles, dispersing the mesoporous silica nanoparticles in phosphate buffer, adding the total arasaponin, carrying out light-shielding oscillation loading, centrifuging, taking solids, rinsing and freeze-drying to obtain the mesoporous silica-loaded total arasaponin, wherein the mass ratio of (500-550) to (3.5-4.5) to (4-5) is calculated as CTAB aqueous solution TEOS= (1-1.2).
  9. 9. The bone healing promoting absorbable bone wax is characterized in that the concentration of the sodium hydroxide aqueous solution is 1.8-2.0M, stirring reaction is carried out for 1.5-2.5 h after TEOS is added, washing is carried out for 2-3 times by using absolute ethyl alcohol and deionized water alternately, calcining is carried out for 5-6 h at a temperature of 1-600 ℃ under a temperature of 550 ℃ per minute, the total saponins of pseudo-ginseng are added according to a proportion that 100-120 mug total saponins of pseudo-ginseng are loaded per milligram of mesoporous silica nanoparticles, the photophobic shaking load is 25-30 ℃ and 80-100 rpm photophobic shaking load is 12-16 h, and the rinsing is carried out by using phosphate buffer solution with pH of 7.0-7.4 pre-cooled at 4-8 ℃.
  10. 10. The method for preparing the absorbable bone wax for promoting bone healing as set forth in claim 1, comprising the steps of: According to the formula mass ratio, uniformly mixing the degradable copolymer and PTMC at 60-65 ℃ to form a base material, reducing the temperature to 45-50 ℃, sequentially adding hydrogenated castor oil, magnesium-calcium silicate-bio-peptide composite microspheres and mesoporous silica-supported pseudo-ginseng total saponins, uniformly mixing to form paste, injecting the paste into a mould at 45-50 ℃, and cooling and shaping at 2-6 ℃ to obtain the bone wax.

Description

Absorbable bone wax for promoting bone healing and preparation method thereof Technical Field The invention belongs to the technical field of bone healing materials, and particularly relates to absorbable bone wax for promoting bone healing and a preparation method thereof. Background Bone wax is a key consumable for controlling bone surface diffuse bleeding in orthopaedics and neurosurgery, and the traditional bone wax takes beeswax and paraffin as core components, has instant hemostatic effect, but can not be degraded, and can form a physical barrier in the residual body to prevent bone cell crawling and bone tissue remodeling, so that bone nonunion, chronic inflammation and infection risks are easily caused, and the application is limited in the surgical scenes of bone healing such as spinal fusion, bone defect repair and the like. In order to break through the limitations of the traditional bone wax, the absorbable bone wax technology is developed gradually, such as a related patent CN105816905B, wherein a polyoxypropylene polyoxyethylene block copolymer and a random copolymer are used as a base material, chitosan is compounded to realize the absorbable and basic healing promotion functions, the CN109125791B introduces strontium-doped hydroxyapatite and microcrystalline cellulose into the copolymer base material to strengthen the bone repair activity, the CN109432487B further optimizes the formula, adopts the block copolymer and the random copolymer as a carrier, and is compounded with the alkylated chitosan and the strontium-doped carbon-containing nano hydroxyapatite to promote the compatibility of the material and bone tissues. Although the prior art realizes the absorbability and the preliminary bone healing promotion capability of bone wax, the method has the obvious technical bottlenecks that firstly the active ingredient has limited healing promotion efficiency, has weak induction effect on proliferation and differentiation of bone cells and has longer bone defect healing period, secondly the substrate and the active ingredient have insufficient synergism, the degradation rate and the bone tissue regeneration rate are difficult to be accurately matched, the problems of hemostasis failure caused by excessively fast degradation or bone growth inhibition caused by excessively slow degradation are easy to occur, and thirdly the function is single, and only hemostasis and basic repair are focused. Along with the development of accurate bone surgery, clinical demands for bone wax are upgraded from hemostasis and absorbability to hemostasis, degradation and high-efficiency osteoinduction. Therefore, the accurate matching of the degradation rate and the bone regeneration is realized by screening high-activity bone induction components, optimizing the proportion of the base material and the active components and the compounding process, so that the bone repair is enhanced, and the limitation of the prior art is broken through. Disclosure of Invention Aiming at the problems of low healing promotion efficiency, insufficient synergy between the base material and the active ingredient, unmatched degradation rate, single function and the like of the bone wax in the prior art, the invention provides the absorbable bone wax for promoting bone healing and the preparation method thereof, the bone wax is prepared by the combination of the degradable copolymer, the magnesium-calcium silicate-biological peptide composite microsphere and mesoporous silica-loaded pseudo-ginseng total saponin, PTMC and hydrogenated castor oil, the limitations of poor degradability and single function of the traditional bone wax are broken through, and the functions of hemostasis, degradation and high-efficiency osteoinduction integration are realized. By the cooperation of the controllable degradation base material and the high-activity functional component, the accurate matching of the degradation rate and the bone regeneration period is achieved. The specific technical scheme is as follows: The absorbable bone wax for promoting bone healing comprises (by mass) a degradable copolymer (25-30), a degradable copolymer (8-10), a degradable copolymer (9-13), a degradable copolymer (3-5) and a degradable copolymer (2-4), PTMC, magnesium-calcium silicate-biological peptide composite microspheres, mesoporous silica-loaded total arasaponin and hydrogenated castor oil; the degradable copolymer is prepared by reacting PEG, stannous octoate, D, L-lactide and epsilon-caprolactone according to the mass ratio of 100 (0.3-0.6) (115-135) at 125-135 ℃ and precipitating in normal hexane, wherein the PEG comprises PEG-400 and PEG-1000 with the mass ratio of (2-4) (5-7); The magnesium-calcium silicate-biological peptide composite microsphere is a porous magnesium-calcium silicate microsphere, biological peptide is subjected to vibration adsorption in a phosphate buffer solution according to the mass ratio of (10-15) (0.8-1.2), solid is obtained by centrifu