CN-121971700-A - Time sequence slow-release temperature-sensitive hydrogel and preparation method and application thereof

Abstract

The invention relates to the technical field of biomedical materials and tissue engineering, and discloses a time sequence slow-release temperature-sensitive hydrogel and a preparation method and application thereof, wherein the preparation method comprises the following steps that a blood sample is subjected to program centrifugation to respectively obtain gel layers of leukocyte-rich platelet fibrin L-PRF and improved platelet-rich fibrin A-PRF, the gel layers are sheared, pre-frozen, freeze-dried, ground and sieved to respectively obtain L-PRF freeze-dried powder and A-PRF freeze-dried powder, and the prepared methacrylic acid gelatin microsphere for encapsulating the L-PRF freeze-dried powder and the microsphere and the A-PRF freeze-dried powder are loaded into a temperature-sensitive hydrogel system formed by chitosan, polygalacturonic acid and beta-glycerophosphate, so that growth factors can be effectively encapsulated in a hydrogel structure and continuously released; the system has better mechanical strength and structural stability, is beneficial to prolonging the retention time of materials in a body and provides stable three-dimensional network support.

Inventors

• ZHOU YANMIN
• CHEN JINGXIA
• LV HUIXIN
• LUO JIAXIN
• WANG YIHAN
• Pei Yangfan
• LIU XIUYU
• CHEN SIYU

Assignees

• 吉林大学

Dates

Publication Date

20260505

Application Date

20260129

Claims (9)

1. 1. The preparation method of the time sequence slow-release temperature-sensitive hydrogel is characterized by comprising the following steps of: Step 1, a blood sample is subjected to procedure centrifugation to respectively obtain gel layers of leukocyte-rich platelet fibrin L-PRF and modified platelet-rich fibrin A-PRF, and the gel layers are sheared, pre-frozen, freeze-dried, ground and sieved to respectively obtain L-PRF freeze-dried powder and A-PRF freeze-dried powder; step 2, emulsifying a methacrylic acid gelatin solution containing a photoinitiator and then carrying out photocrosslinking to obtain methacrylic acid gelatin microspheres; step 3, adding the L-PRF freeze-dried powder into a methacrylic acid gelatin solution containing a photoinitiator, and emulsifying and photo-crosslinking to obtain methacrylic acid gelatin microspheres coated with the L-PRF freeze-dried powder; Step 4, performing ultrasonic treatment on the chitosan solution and the polygalacturonic acid solution to form a polyelectrolyte complex suspension, and adjusting the pH value of the polyelectrolyte complex suspension by adding an alkaline solution; and 5, adding A-PRF freeze-dried powder and methacrylic acid gelatin microspheres coated with L-PRF freeze-dried powder into the polyelectrolyte complex suspension, dropwise adding beta-sodium glycerophosphate solution, stirring and incubating under a low temperature condition to obtain the time sequence slow-release temperature-sensitive hydrogel carrying the growth factors.
2. 2. The method for preparing a time-series slow-release temperature-sensitive hydrogel according to claim 1, wherein the L-PRF in the step 1 is obtained by centrifugation at 3000r/min for 10: 10min, and the A-PRF is obtained by centrifugation at 1300: 1300 r/min for 8: 8 min.
3. 3. The method for preparing a time-series slow-release temperature-sensitive hydrogel according to claim 1, wherein in step 2, a methacrylic acid gelatin solution containing a photoinitiator is dripped into 37 ℃ liquid paraffin containing an emulsifier, and stirred under the water bath condition of 37 ℃ to form an emulsion, and then the emulsion is placed under the ice bath condition to be continuously stirred; and irradiating the emulsion obtained by emulsification under a 405 nm light source for 5min photo-crosslinking polymerization, and adding acetone to wash after the photo-crosslinking polymerization is finished so as to remove an oil phase.
4. 4. The method for preparing a time-series slow-release temperature-sensitive hydrogel according to claim 1, wherein in step 3, the L-PRF freeze-dried powder is added into a methacrylic acid gelatin solution containing a photoinitiator and mixed, the mixed methacrylic acid gelatin solution is dripped into 37 ℃ liquid paraffin containing an emulsifier, the mixture is stirred under the water bath condition of 37 ℃ to form an emulsion, and then the emulsion is placed under the ice bath condition to be continuously stirred; and irradiating the emulsion obtained by emulsification under a 405 nm light source for 5min photo-crosslinking polymerization, and adding acetone to wash after the photo-crosslinking polymerization is finished so as to remove an oil phase.
5. 5. The method for preparing the time-series slow-release temperature-sensitive hydrogel according to claim 1, wherein the particle sizes of the methacrylic acid gelatin microsphere obtained in the step 2 and the methacrylic acid gelatin microsphere coated with the L-PRF freeze-dried powder obtained in the step 3 are 90-170 μm.
6. 6. The method for preparing the time-series slow-release temperature-sensitive hydrogel according to claim 1, wherein in the step 5, stirring and incubation under the low-temperature condition are specifically performed under the ice bath condition for 20 minutes and under the 4 ℃ condition.
7. 7. A time sequence slow-release temperature-sensitive hydrogel, which is characterized by being prepared by the preparation method of any one of claims 1-6.
8. 8. The use of the time-series slow-release temperature-sensitive hydrogel according to claim 7 for preparing a bone regeneration material for bone defect cavity vascularization after maxillary sinus lifting operation.
9. 9. The use according to claim 8, wherein the time-series slow-release temperature-sensitive hydrogel fills a cavity formed after maxillary sinus lifting operation in an injection manner.

Description

Time sequence slow-release temperature-sensitive hydrogel and preparation method and application thereof Technical Field The invention relates to the technical field of biomedical materials and tissue engineering, in particular to a time sequence slow-release temperature-sensitive hydrogel and a preparation method and application thereof. Background In the process of oral implant treatment, especially in the process of implant in the maxillary posterior dental region, the maxillary sinus lifting operation is often required due to insufficient alveolar bone absorption or bone mass so as to realize stable implant of the implant. The procedure provides room for subsequent bone tissue regeneration by lifting the maxillary sinus floor mucosa to form a new osteogenic cavity. However, the osteogenic cavity is usually surrounded by a hard bone wall and a flexible mucosa, and in a physiological state, the maxillary sinus mucosa may perform micro motion along with respiratory activity, resulting in a decrease in stability of the cavity, thereby affecting the bone tissue regeneration effect. In addition, the cavity formed after the maxillary sinus lifting operation is often irregular, the material is easy to loose, shift or collapse, and the occurrence risk of postoperative complications is increased. Blood vessels serve as key structures for the transport of oxygen, nutrients, removal of metabolic waste products and transmission of biological signals, and play an important role in the bone regeneration process. Angiogenesis is closely related to the osteogenic process, and once the number of vascular endothelial cells is reduced or function is compromised, signal communication between the bone matrix and the blood vessels is affected, resulting in reduced angiogenic capacity of the bone, further limiting new bone formation. Therefore, in the repair of maxillary sinus bone defects, a biomaterial capable of adapting to the morphology of irregular bone defect cavities and effectively promoting angiogenesis is needed. The missing teeth not only seriously affect the maxillofacial appearance of a patient, but also have adverse effects on functions such as chewing, pronunciation and the like, especially the missing teeth in a rear tooth area easily cause the reduction of the chewing function, increase the burden of a digestive system and further affect the overall health of the patient. The long-term missing teeth can also cause the vaporization aggravation of the maxillary sinus and the vertical absorption of the alveolar bone, so that the height of the residual bone in the posterior tooth area is gradually reduced, and the difficulty of subsequent planting and repairing is obviously increased. Currently, bone formation is often promoted clinically by maxillary sinus lifting in combination with bone replacement materials. Wherein, the elastic maxillary sinus mucosa (maxillary sinus membrane, MSM) can form a surrounding structure with the hard bone wall to provide a space for osteogenesis. However, the lacunae morphology is often relatively irregular, and common filling materials are difficult to adapt individually to complex bone defect areas, which easily results in loosening, displacement or poor bone healing of the materials in the body. In existing osteogenic repair strategies, bone tissue engineering generally relies on the synergistic effects of biomaterial scaffolds, bioactive factors, and seed cells. However, loading living cells in biological scaffolds still faces challenges such as limited cell sources, complex in-vitro expansion processes, and difficult maintenance of cell activity and function. In contrast, cell-free biomaterials have certain advantages in advancing the clinical transformation of bone regeneration materials, and the strategy is to induce and regulate the migration, proliferation and differentiation of host cells by the physical and chemical properties of the materials themselves, thereby achieving tissue regeneration. In addition, the hydrogel material has wide application prospect in the field of tissue engineering due to good biocompatibility, injectability and plasticity. The hydrogel can be injected into a body in a liquid state by reasonably designing the composition and structure of the hydrogel, and is converted into a gel state in situ under physiological conditions, so that surgical wounds can be reduced to a certain extent and irregular defect cavities can be adapted. However, how to achieve both a good adaptation to the morphology of the bone defect, an effective promotion of angiogenesis and a sustained support for the osteogenesis process in a hydrogel system remains a problem to be solved in the prior art. Clinically, a commonly used filling material after maxillary sinus lifting operation includes deproteinized bovine bone mineral (deproteinized bovine bone mineral, DBBM), which has a composition similar to that of human bone tissue, and can provide bone supporting effect to some