A Janus-structured bio-based polyurethane membrane for inducing periodontal tissue regeneration

Abstract

The invention discloses a Janus structure bio-based polyurethane film for inducing periodontal tissue regeneration, belonging to the technical field of medical materials. The Janus structure bio-based polyurethane film is synthesized from the following main raw materials: 100 parts of hydrophilic diol, 100 parts of hydrophobic diol, 22.2-44.4 parts of isophorone diisocyanate, 22.6-45.2 parts of lysine diisocyanate, 5.9-17.8 parts of chain extender E1 containing tertiary amine structure, and 100 parts of hydrophobic diol. 5.9-17.8 parts, monohalogenated alkane 8.2-24.7 parts, 3,4-dihydroxybenzoic acid 7.7-23.1 parts, organic bismuth catalyst 0.4 parts, prepared by synthesizing bio-based polyurethane and casting into film; the prepared Janus structure bio-based polyurethane film shows the characteristics of one side being hydrophilic and the other side being hydrophobic; the quaternary ammonium salt on the hydrophilic side can hinder cell migration and inhibit bacterial growth, provide a sterile environment, and the functional groups on the hydrophobic side can promote periodontal tissue regeneration. At the same time, the bio-based polyurethane film has good biocompatibility and degradability.

Description

Technical Field

[0001] The invention belongs to the technical field of medical materials, and in particular relates to a Janus structure bio-based polyurethane film for inducing periodontal tissue regeneration. Background Art

[0002] Periodontitis is a common chronic infectious disease of the oral cavity. The most important pathological feature is the destruction of periodontal supporting tissues. In severe cases, it can lead to loosening and detachment of teeth. The purpose of periodontal treatment is to control plaque, eliminate inflammation, achieve regeneration and repair of defective tissues, and maintain periodontal health. Periodontal tissues include alveolar bone, cementum, and periodontal ligament. They have complex structures, diverse layers, and contain soft and hard tissue structures. Their repair requires the realization of multi-tissue regeneration of the "alveolar bone-periodontal ligament-cementum" physiological structure, which is the focus and difficulty in the field of oral research. At present, guided tissue regeneration technology (Guided tissue regeneration, GTR) is widely used in the clinical treatment of periodontal defects and has achieved certain therapeutic effects, but it still cannot achieve the goal of periodontal multi-tissue regeneration to restore the physiological structure of periodontal tissues. Therefore, periodontal multi-tissue regeneration is still an urgent problem to be solved in oral clinics.

[0003] In recent years, with the development of stomatology, materials for inducing periodontal tissue regeneration have received more and more attention. For example, patent CN 118059301 A discloses a method for preparing a dual-function membrane for periodontal bone tissue repair. After polarization treatment, the material is more compatible with the piezoelectric properties of human bones, that is, it has a piezoelectric coefficient that can fully exert the osteogenesis effect. Then, under the stimulation of an ultrasonic device, the piezoelectric signal is stimulated to promote the dual effects of antibacterial and bone regeneration, and promote periodontal tissue regeneration quickly and efficiently. However, the invention still faces problems such as high price and complex process. Summary of the invention

[0004] In view of the current state of the art, the present invention provides a Janus structure bio-based polyurethane membrane for inducing periodontal tissue regeneration and a preparation method thereof. The membrane has the characteristics of simple preparation, stable properties and good biocompatibility; it is easy to use, has antibacterial and sterilization properties, is cheap and safe, and promotes tooth bone regeneration while fixing fibroblasts.

[0005] To achieve the above-mentioned purpose and other related purposes, the present invention provides a Janus structure bio-based polyurethane membrane for inducing periodontal tissue regeneration, wherein the Janus structure bio-based polyurethane membrane for inducing periodontal tissue regeneration is prepared by synthesis of the following main components, and the mass fraction of each component is:

[0006] 100 parts of hydrophilic glycol;

[0007] 100 parts of hydrophobic glycol;

[0008] 22.2-44.4 parts of hydrophobic diisocyanate;

[0009] 22.6-45.2 parts of hydrophilic diisocyanate;

[0010] 5.9-17.8 parts of chain extender E1 containing tertiary amine structure;

[0011] 5.9-17.8 parts of chain extender E2 containing tertiary amine structure;

[0012] 8.2-24.7 parts of monohalogenated alkanes;

[0013] 3,4-dihydroxybenzoic acid 7.7-23.1 parts;

[0014] 0.4 parts of organic bismuth catalyst;

[0015] Preferably, the hydrophobic diisocyanate is isophorone diisocyanate.

[0016] Preferably, the hydrophilic diisocyanate is lysine diisocyanate.

[0017] Preferably, the chain extender E1 containing a tertiary amine structure is a compound having the following structural formula: (1);

[0018] In formula (1), n=1-4.

[0019] Preferably, the chain extender E2 containing a tertiary amine structure is a compound having the following structural formula: (2);

[0020] In formula (2), R is one of methyl, ethyl and isopropyl.

[0021] Preferably, the monohalogenated alkane is a compound comprising the following structural formula: (3);

[0022] In formula (3), m=4 or 6 or 8, and X is Br or I.

[0023] Preferably, the hydrophilic diol includes at least one of hydroxy-terminated polybutadiene and polyethylene glycol.

[0024] Preferably, the hydrophobic diol includes at least one of biodegradable polycaprolactone diol, polydimethylsiloxane diol, polytetramethylene glycol, polylactic acid diol and polypropylene glycol.

[0025] Preferably, the diol number average molecular weight is M n =1000-4000 g / mol.

[0026] The invention provides a Janus structure bio-based polyurethane film for inducing periodontal tissue regeneration and its application in the field of dental implantation.

[0027] The present invention also provides a method for preparing a Janus structure bio-based polyurethane film for inducing periodontal tissue regeneration, comprising the following steps:

[0028] The hydrophilic diol and the hydrophobic diol are dried at 110-130° C. and -0.1 MPa for 1-2 hours, and cooled to room temperature to obtain the dehydrated hydrophilic diol and the hydrophobic diol;

[0029] The dehydrated hydrophilic diol and the hydrophobic diol are poured into a reactor, isophorone diisocyanate, lysine diisocyanate and 0.4 parts of an organic bismuth catalyst are added, and the mixture is stirred at 90-95° C. to be uniformly mixed and reacted for 2-3 hours to obtain a bio-based polyurethane prepolymer;

[0030] The bio-based polyurethane prepolymer is cooled to 70-80° C., and a chain extender E2 containing a tertiary amine structure and a monohalogenated alkane are added to carry out chain extension and quaternization reaction for 2-3 hours to obtain a monofunctional bio-based polyurethane slurry;

[0031] Adding chain extender E1 to the above monofunctional bio-based polyurethane slurry, reacting at 70-80° C. for 1-2 hours, cooling to 50-60° C., adding 3,4-dihydroxybenzoic acid and reacting for 1-2 hours to obtain a bifunctional bio-based polyurethane slurry;

[0032] Finally, the above-mentioned bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, and immersed in deionized water. After being placed in a 60-100°C oven for 24-48 hours, the migration process of the hydrophilic chain and the hydrophilic group was completed to obtain a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration.

[0033] The beneficial effects of the present invention are:

[0034] 1. The bio-based polyurethane prepared by the present invention using terminal hydroxyl polybutadiene and polycaprolactone diol as soft segments and lysine diisocyanate as hard segments can be rapidly degraded in the presence of enzymes.

[0035] 2. The Janus structure bio-based polyurethane membrane prepared by the present invention not only has the characteristics of traditional GBR (guided bone regeneration) materials in promoting tooth bone regeneration, but also can provide a sterile environment for the affected area by inhibiting bacterial growth and preventing the adverse consequences of fibroblast migration.

[0036] 3. The Janus structure bio-based polyurethane film prepared by the present invention has a simple production process and low material price.

[0037] 4. The raw materials used in the present invention are bio-based and are new materials with sustainable development. The raw materials are non-toxic and harmless to organisms and are suitable for medical materials. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0039] Figure 1 The present invention is a flow chart of the preparation method of the Janus structure bio-based polyurethane film;

[0040] Figure 2 The mechanical properties of the bio-based polyurethane film of the Janus structure of the present invention;

[0041] Figure 3 The water contact angle data of the bio-based polyurethane film with Janus structure of the present invention;

[0042] Figure 4 This is a picture of the water contact angle of the bio-based polyurethane film with a Janus structure in Example 2 of the present invention;

[0043] Figure 5 The antibacterial data of the bio-based polyurethane film of the Janus structure of the present invention;

[0044] Figure 6 In vitro cell compatibility data of the bio-based polyurethane membrane of the Janus structure of the present invention;

[0045] Figure 7 Degradation data of the bio-based polyurethane film of the Janus structure of the present invention in an enzyme environment;

[0046] Figure 8 The results of alkaline phosphatase activity of MC3T3-E1 osteoblasts after culturing on the Janus structure bio-based polyurethane membrane surface for 7 days and 14 days;

[0047] Fig. 9 The extracellular matrix mineralization of MC3T3-E1 osteoblasts in Examples 1-5 and Comparative Examples 1-3 after being cultured on the surface of the Janus-structured bio-based polyurethane membrane for 7 days and 14 days. DETAILED DESCRIPTION

[0048] The following is an explanation of the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the features in the following embodiments and the embodiments can be combined with each other without conflict. It should also be understood that the terms used in the embodiments of the present invention are intended to describe specific embodiments, rather than to limit the scope of protection of the present invention. The test methods for which specific conditions are not specified in the following examples are usually carried out under conventional conditions or according to the conditions recommended by the manufacturers.

[0049] It should be noted that the terms such as "upper", "lower", "left", "right", "middle" and "one" etc. used in this specification are only for the convenience of description and are not intended to limit the scope of implementation of the present invention. Changes or adjustments to their relative relationships, without substantially changing the technical content, should also be regarded as the scope of implementation of the present invention.

[0050] The invention provides a Janus structure bio-based polyurethane film for inducing periodontal tissue regeneration. The Janus structure bio-based polyurethane film for inducing periodontal tissue regeneration comprises the following components, and the weight proportions of the components are: 100 parts of hydrophilic diol, 100 parts of hydrophobic diol, 22.2-44.4 parts of hydrophobic diisocyanate, 22.6-45.2 parts of hydrophilic diisocyanate, 5.9-17.8 parts of chain extender E1 containing tertiary amine structure, 5.9-17.8 parts of chain extender E2 containing tertiary amine structure, 8.2-24.7 parts of monohalogenated alkane, 7.7-23.1 parts of 3,4-dihydroxybenzoic acid and 0.4 parts of organic bismuth catalyst.

[0051] The hydrophilic diol includes at least one of terminal hydroxyl polybutadiene and polyethylene glycol, that is, the hydrophilic diol can be selected from any one of the above-mentioned types, such as terminal hydroxyl polybutadiene or polyethylene glycol; the hydrophilic diol can also be any two combinations of the above-mentioned types, such as a combination of terminal hydroxyl polybutadiene and polyethylene glycol, which are not listed here one by one. If the hydrophilic diol is a combination of two types, there is no restriction on the ratio between the components in the combination, and they can be mixed in any ratio.

[0052] Wherein, the hydrophobic diol includes at least one of biodegradable polycaprolactone diol, polydimethylsiloxane diol, polypropylene glycol, polytetrahydrofuran diol, and polylactic acid diol, that is, the hydrophobic diol can be selected from any one of the above-mentioned types, such as biodegradable polycaprolactone diol, polydimethylsiloxane diol or polypropylene glycol or polytetrahydrofuran diol or polylactic acid diol; the hydrophobic diol can also be any two or three compositions listed above, such as a composition of biodegradable polycaprolactone diol and polydimethylsiloxane diol or a composition of biodegradable polycaprolactone diol, polydimethylsiloxane diol, and polypropylene glycol, which are not listed here one by one. If the hydrophobic diol is a composition of two or three, the ratio between the components in the composition is not limited, and it can be mixed in any proportion.

[0053] The hydrophilic diisocyanate includes at least one of lysine diisocyanate and hexamethylene diisocyanate. That is, the hydrophilic diisocyanate can be selected from any one of the above-mentioned types, such as lysine diisocyanate and hexamethylene diisocyanate. The aliphatic diisocyanate can also be a composition of lysine diisocyanate and hexamethylene diisocyanate. When the aliphatic diisocyanate is a composition, the ratio of lysine diisocyanate and hexamethylene diisocyanate is not limited, and lysine diisocyanate and hexamethylene diisocyanate can be mixed in any proportion. In the present invention, the weight percentage of the hydrophilic diisocyanate is 22.6-45.2 parts, for example, the weight percentage of the aliphatic diisocyanate can be any value within the above range such as 22.6 parts, 33.9 parts or 45.2 parts.

[0054] The hydrophobic diisocyanate includes at least one of isophorone diisocyanate, dicyclohexylmethane diisocyanate, and 1,4-cyclohexane diisocyanate. That is, the alicyclic diisocyanate can be selected from any one of the above-mentioned types, such as isophorone diisocyanate and 1,4-cyclohexane diisocyanate, and the alicyclic diisocyanate can also be a combination of any two or three of the above-mentioned types, such as a combination of isophorone diisocyanate and dicyclohexylmethane diisocyanate, or a combination of isophorone diisocyanate, dicyclohexylmethane diisocyanate and 1,4-cyclohexane diisocyanate. They are not listed here one by one. If the hydrophobic diisocyanate is a combination of two or three types, there is no restriction on the ratio between the components in the composition, and they can be mixed in any proportion. In the present invention, the weight percentage of the alicyclic diisocyanate is 22.2-44.4 parts, for example, the weight percentage of the alicyclic diisocyanate can be any value within the above range, such as 22.2 parts, 33.3 parts or 44.4 parts.

[0055] The number average molecular weight of the hydrophilic diol and the hydrophobic diol used in all the following examples and comparative examples is M n=2000 g / mol.

[0056] The chain extender E1 containing a tertiary amine structure is at least one of N-methyldiethanolamine, N-ethyldiethanolamine, N-propyldiethanolamine and N-butyldiethanolamine, that is, the small molecule chain extender E1 can be selected from any one of the above-listed types, and the small molecule chain extender E1 can also be a combination of N-methyldiethanolamine, N-ethyldiethanolamine, N-propyldiethanolamine and N-butyldiethanolamine. In the present invention, the weight portion of the small molecule chain extender E1 is 5.9-17.8 parts, for example, it can be any value within the above range such as 5.9 parts, 11.8 parts, 17.8 parts, etc.

[0057] The tertiary amine structure chain extender E2 includes at least one of 3-dimethylamino-1,2-propylene glycol, 3-diethylamino-1,2-propylene glycol, and 3-diisopropylamino-1,2-propylene glycol, that is, the tertiary amine structure chain extender E2 can be selected from any one of the types listed above, such as 3-dimethylamino-1,2-propylene glycol, 3-diethylamino-1,2-propylene glycol, and 3-diisopropylamino-1,2-propylene glycol. The tertiary amine structure chain extender E2 can also be a mixture of two or three of the types listed above, such as a combination of 3-dimethylamino-1,2-propylene glycol and 3-diethylamino-1,2-propylene glycol, or a combination of 3-dimethylamino-1,2-propylene glycol, 3-diethylamino-1,2-propylene glycol, and 3-diisopropylamino-1,2-propylene glycol, which are not listed here one by one. In the present invention, the weight proportion of the chain extender E2 having a tertiary amine structure is 5.9-17.8 parts, for example, it can be any value within the above range such as 5.9 parts, 11.8 parts, 17.8 parts, etc.

[0058] See also Figure 1 The present invention provides a method for preparing a Janus structure bio-based polyurethane film for inducing periodontal tissue regeneration, comprising the following steps:

[0059] S1: drying the hydrophilic diol and the hydrophobic diol at 110-130° C. and -0.1 MPa for 1-2 h, and cooling to room temperature to obtain the dehydrated hydrophilic diol and the hydrophobic diol;

[0060] For example, the dehydration temperature of the diol can be any value within the above temperature range, such as 110° C., 120° C. or 130° C., and the drying time can be any value within the above drying time range, such as 1 h, 1.5 h or 2 h.

[0061] S2: pouring the above-mentioned hydrophilic diol and hydrophobic diol from which water has been removed into a reactor, adding isophorone diisocyanate, lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring at 90-95° C. to mix them evenly and reacting for 2-3 hours to obtain a bio-based polyurethane prepolymer;

[0062] For example, the temperature of the diol and the isocyanate can be any value within the above temperature range, such as 90°C, 93°C or 95°C, and the reaction time can be any value within the above drying time range, such as 2h, 2.5h or 3h.

[0063] S3: cooling the bio-based polyurethane prepolymer to 70-80° C., adding a chain extender E2 containing a tertiary amine structure and a monohalogenated alkane, and performing chain extension and quaternization reactions for 2-3 hours to obtain a monofunctional bio-based polyurethane slurry;

[0064] For example, the chain extension and quaternization reaction temperature can be any value within the above temperature range, such as 70°C, 75°C or 80°C, and the reaction time can be any value within the above drying time range, such as 2h, 2.5h or 3h.

[0065] S4: adding a chain extender E1 to the monofunctional bio-based polyurethane slurry, reacting at 70-80° C. for 1-2 hours, cooling to 50-60° C., adding 3,4-dihydroxybenzoic acid and reacting for 1-2 hours to obtain a bifunctional bio-based polyurethane slurry;

[0066] For example, the second chain extension and quaternization reaction temperatures can be any value within the above temperature ranges of 70°C, 75°C or 80°C and 50°C, 55°C or 60°C, respectively, and the reaction time can be any value within the above drying time ranges of 1h, 1.5h or 2h.

[0067] S5Finally, the bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, and then immersed in deionized water and placed in a 60-100°C oven for 24-48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration;

[0068] For example, the curing temperature may be any value within the above temperature range, such as 60° C., 80° C. or 100° C., and the curing time may be any value within the above drying time range, such as 24 h, 36 h or 48 h.

[0069] The technical scheme of the present invention is described in detail below through several specific examples. Unless otherwise specified, the raw materials and reagents used in the following examples are all commercially available products, or can be prepared by conventional methods in the art, and the instruments used in the examples are all commercially available. Example 1

[0070] (1) drying the terminal hydroxyl polybutadiene and polycaprolactone diol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain the dehydrated terminal hydroxyl polybutadiene and polycaprolactone diol;

[0071] (2) pouring 100 parts of the above-mentioned dehydrated hydroxy-terminated polybutadiene and 100 parts of polycaprolactone diol into a reactor, adding 22.2 parts of isophorone diisocyanate, 22.6 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95°C for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0072] (3) Cooling the above bio-based polyurethane prepolymer to 80°C, adding 5.9 parts of a chain extender E2 containing a tertiary amine structure in which R is methyl and 8.2 parts of a monohalogenated alkane in which m=6 and X is Br, and performing chain extension and quaternization reaction for 2 hours to obtain a monofunctional bio-based polyurethane slurry;

[0073] (4) Add 5.9 parts of chain extender E1 (n=1) to the above monofunctional bio-based polyurethane slurry, react at 80°C for 2 hours, cool to 60°C, add 7.7 parts of 3,4-dihydroxybenzoic acid and react for 1 hour to obtain a bifunctional bio-based polyurethane slurry;

[0074] (5) Finally, the bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in an oven at 80°C for 48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration. Example 2

[0075] (1) drying the terminal hydroxyl polybutadiene and polycaprolactone diol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain the dehydrated terminal hydroxyl polybutadiene and polycaprolactone diol;

[0076] (2) pouring 100 parts of the above-mentioned dehydrated hydroxy-terminated polybutadiene and 100 parts of polycaprolactone diol into a reactor, adding 44.4 parts of isophorone diisocyanate, 45.2 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95°C for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0077] (3) Cooling the above bio-based polyurethane prepolymer to 80°C, adding 17.8 parts of a chain extender E2 containing a tertiary amine structure in which R is methyl and 24.7 parts of a monohalogenated alkane in which m=6 and X is Br, and performing chain extension and quaternization reaction for 2 hours to obtain a monofunctional bio-based polyurethane slurry;

[0078] (4) Add 17.8 parts of chain extender E1 (n=1) to the above monofunctional bio-based polyurethane slurry, react at 80°C for 2 hours, cool to 60°C, add 23.1 parts of 3,4-dihydroxybenzoic acid and react for 1 hour to obtain a bifunctional bio-based polyurethane slurry;

[0079] (5) Finally, the bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in an oven at 80°C for 24 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration. Example 3

[0080] (1) drying polyethylene glycol and polypropylene glycol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain dehydrated polyethylene glycol and polypropylene glycol;

[0081] (2) pouring 100 parts of the above-mentioned dehydrated polyethylene glycol and 100 parts of polypropylene glycol into a reactor, adding 44.4 parts of isophorone diisocyanate, 45.2 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95° C. for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0082] (3) Cooling the above bio-based polyurethane prepolymer to 80°C, adding 17.8 parts of a chain extender E2 containing a tertiary amine structure in which R is methyl and 24.7 parts of a monohalogenated alkane in which m=6 and X is Br, and performing chain extension and quaternization reaction for 2 hours to obtain a monofunctional bio-based polyurethane slurry;

[0083] (4) Add 17.8 parts of chain extender E1 (n=1) to the above monofunctional bio-based polyurethane slurry, react at 80°C for 2 hours, cool to 60°C, add 23.1 parts of 3,4-dihydroxybenzoic acid and react for 1 hour to obtain a bifunctional bio-based polyurethane slurry;

[0084] (5) Finally, the bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in an oven at 80°C for 48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration. Example 4

[0085] (1) drying the terminal hydroxyl polybutadiene and polycaprolactone diol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain the dehydrated terminal hydroxyl polybutadiene and polycaprolactone diol;

[0086] (2) pouring 100 parts of the above-mentioned dehydrated hydroxy-terminated polybutadiene and 100 parts of polycaprolactone diol into a reactor, adding 44.4 parts of isophorone diisocyanate, 45.2 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95°C for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0087] (3) Cooling the above bio-based polyurethane prepolymer to 80° C., adding 17.8 parts of a chain extender E2 containing a tertiary amine structure in which R is methyl and 24.7 parts of a monohalogenated alkane in which m=4 and X is Br, and performing chain extension and quaternization reaction for 2 hours to obtain a monofunctional bio-based polyurethane slurry;

[0088] (4) Add 17.8 parts of chain extender E1 (n=4) to the above monofunctional bio-based polyurethane slurry, react at 80°C for 2 hours, cool to 60°C, add 23.1 parts of 3,4-dihydroxybenzoic acid and react for 1 hour to obtain a bifunctional bio-based polyurethane slurry;

[0089] (5) Finally, the bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in an oven at 80°C for 48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration. Example 5

[0090] (1) drying the terminal hydroxyl polybutadiene and polycaprolactone diol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain the dehydrated terminal hydroxyl polybutadiene and polycaprolactone diol;

[0091] (2) pouring 100 parts of the above-mentioned dehydrated hydroxy-terminated polybutadiene and 100 parts of polycaprolactone diol into a reactor, adding 44.4 parts of isophorone diisocyanate, 45.2 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95°C for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0092] (3) Cooling the above bio-based polyurethane prepolymer to 80°C, adding 17.8 parts of a chain extender E2 containing a tertiary amine structure in which R is an isopropyl group and 24.7 parts of a monohalogenated alkane in which m=6 and X is Br, and performing chain extension and quaternization reaction for 2 hours to obtain a monofunctional bio-based polyurethane slurry;

[0093] (4) Add 17.8 parts of chain extender E1 (n=1) to the above monofunctional bio-based polyurethane slurry, react at 80°C for 2 hours, cool to 60°C, add 23.1 parts of 3,4-dihydroxybenzoic acid and react for 1 hour to obtain a bifunctional bio-based polyurethane slurry;

[0094] (5) Finally, the bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in an oven at 80°C for 48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration.

[0095] Comparative Example 1

[0096] (1) drying the terminal hydroxyl polybutadiene and polycaprolactone diol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain the dehydrated terminal hydroxyl polybutadiene and polycaprolactone diol;

[0097] (2) pouring 100 parts of the above-mentioned dehydrated hydroxy-terminated polybutadiene and 100 parts of polycaprolactone diol into a reactor, adding 44.4 parts of isophorone diisocyanate, 45.2 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95°C for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0098] (3) The bio-based polyurethane prepolymer was cooled to 80°C, 35.6 parts of a chain extender E2 containing a tertiary amine structure in which R is methyl and 49.4 parts of a monohalogenated alkane in which m=6 and X is Br were added, and the chain extension and quaternization reaction was carried out for 2 hours to obtain a monofunctional bio-based polyurethane slurry;

[0099] (4) Finally, the monofunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in an oven at 80°C for 48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration.

[0100] Comparative Example 2

[0101] (1) drying the terminal hydroxyl polybutadiene and polycaprolactone diol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain the dehydrated terminal hydroxyl polybutadiene and polycaprolactone diol;

[0102] (2) pouring 100 parts of the above-mentioned dehydrated hydroxy-terminated polybutadiene and 100 parts of polycaprolactone diol into a reactor, adding 44.4 parts of isophorone diisocyanate, 45.2 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95°C for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0103] (3) Add 17.8 parts of chain extender E1 (n=1) to the above bio-based polyurethane prepolymer, react at 80°C for 2 hours, cool to 60°C, add 23.1 parts of 3,4-dihydroxybenzoic acid and react for 1 hour to obtain a monofunctional bio-based polyurethane elastomer slurry;

[0104] (4) Finally, the monofunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in an oven at 80°C for 48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration.

[0105] Comparative Example 3

[0106] (1) drying the terminal hydroxyl polybutadiene and polycaprolactone diol at 110° C. and −0.1 MPa for 2 h, and cooling to room temperature to obtain the dehydrated terminal hydroxyl polybutadiene and polycaprolactone diol;

[0107] (2) pouring 100 parts of the above-mentioned dehydrated hydroxy-terminated polybutadiene and 100 parts of polycaprolactone diol into a reactor, adding 44.4 parts of isophorone diisocyanate, 45.2 parts of lysine diisocyanate and 0.4 parts of an organic bismuth catalyst, stirring and reacting at 95°C for 3 hours, mixing evenly, and obtaining a bio-based polyurethane prepolymer;

[0108] (3) Cooling the above bio-based polyurethane prepolymer to 80° C., adding 17.8 parts of a chain extender E2 containing a tertiary amine structure in which R is a methyl group, and carrying out a chain extension reaction for 2 hours to obtain a bio-based polyurethane slurry;

[0109] (4) adding 17.8 parts of chain extender E1 (n=1) to the above bio-based polyurethane slurry, reacting at 80° C. for 2 h to obtain a bio-based polyurethane slurry after the reaction is completed;

[0110] (5) Finally, the above-mentioned bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, and then immersed in deionized water and placed in an 80°C oven for 48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration.

[0111] Test Case

[0112] Mechanical test: The mechanical properties of the Janus structure bio-based polyurethane films prepared in Examples 1-5 and Comparative Examples 1-3 were tested, and the tensile strength, elongation at break and other tests were performed in accordance with relevant standards. The specific test method is as follows: The mechanical properties of the samples were tested using a CMT4304 universal testing machine, with a tensile speed of 100 mm / min, and each polyurethane elastomer sample was tested three times in parallel at different positions, and the average value was taken.

[0113] result Figure 2 As shown, from Figure 2 The test results show that the bio-based polyurethane elastomers prepared in Examples 1-5 have good mechanical properties, and the bio-based polyurethane elastomers prepared in Comparative Examples 1-3 have only monofunctional quaternary ammonium groups or no quaternary ammonium groups during the preparation process. Compared with the bio-based polyurethane elastomers prepared in Comparative Examples 1-3 in Examples 1-5, the mechanical properties of the polyurethane elastomers prepared in Comparative Examples 1-3 are reduced, indicating that the bifunctional quaternary ammonium groups can improve the mechanical properties of the polyurethane elastomer.

[0114] Water contact angle test: The Janus structure bio-based polyurethane films prepared in Examples 1-5 and Comparative Examples 1-3 were tested for water contact angle. The water contact angles on both sides of the bio-based polyurethane were tested using an OCA 20 water contact angle meter. The test medium was deionized water, and the volume of the medium dropped each time was set to 1.0 μL. The water contact angles of each group of samples were tested in parallel 3 times at different positions and the average value was taken. All samples were water-induced. The results are shown in Figure 2. Figure 3 shown.

[0115] from Figure 3 The test results show that the bio-based polyurethane elastomers prepared in Examples 1-5 have the characteristics of being hydrophobic on one side and hydrophilic on the other side, proving that they have a Janus structure. The bio-based polyurethane elastomers prepared in Comparative Examples 1-3 have only monofunctional quaternary ammonium salt groups or no quaternary ammonium salt groups during the preparation process. Therefore, they present the same hydrophilic and hydrophobic properties on both sides. Figure 4 Provided are photos of the water contact angles on both sides of the polyurethane before and after water induction in Example 2.

[0116] Antibacterial test: The Janus structure bio-based polyurethane membranes prepared in Examples 1-5 and Comparative Examples 1-3 were subjected to antibacterial tests. LB agar medium was prepared and then sterilized by high pressure. Bacteria were revived at a ratio of 1:1000 and then placed on a shaker at 37°C and 120 rpm overnight. Bacteria were cultured and the material and bacterial solution were mixed at 1×10 6 The concentration of 100000 was co-cultured and placed on a shaker. The conditions were also set to 120 rpm and 37°C. Plate the successfully cultured bacterial solution, dilute it 10,000 times, and spread it on LB agar medium. Take 5 μL for each colony. Then place it in a 37-degree incubator, culture it for 12 hours, take pictures and count, and calculate the antibacterial rate.

[0117] from Figure 5 It can be seen from the test results that the bio-based polyurethane elastomers prepared in Examples 1-5 all have excellent antibacterial properties, which is due to the rich quaternary ammonium salts with antibacterial effects in the molecular chain. The bio-based polyurethane elastomers prepared in Comparative Examples 1-3 only have monofunctional quaternary ammonium salt groups or no quaternary ammonium salt groups during the preparation process, so they show weak antibacterial effects, and Comparative Example 3 does not even have an antibacterial effect.

[0118] In vitro biocompatibility test: The Janus structure bio-based polyurethane membranes prepared in Examples 1-5 and Comparative Examples 1-3 were subjected to in vitro biocompatibility tests. Mouse embryonic fibroblasts (NIH / 3T3) and MEM complete medium containing 10% fetal bovine serum were used for culture. The growth of NIH / 3T3 cells in the culture flask was observed under an optical microscope. After the cell density reached about 80%, the old culture medium was poured onto the clean bench, sterile PBS solution was added for rinsing, the residual culture medium was removed, and then 1 mL of EDTA-containing trypsin was added and digested at room temperature for 1 min. Use a microscope to check whether the cells are completely absorbed, and then add 1 mL of fresh culture medium to terminate the digestion. The cell suspension was then centrifuged at 1500 rpm / min for 5 minutes. The supernatant was discarded, the cells were resuspended in fresh culture medium, and the cell concentration was diluted to 5×10 after counting using a cell counting plate. 4 cells / mL. The mixture was then evenly added to a 96-well plate, 200 μL was added to each well, and the plate was poured in a CO2 cell culture incubator for 24 h. The culture medium containing the sample extract was added to the cell wells of the experimental group (the sample extract (20 mg / ml) was a sterile extract obtained after being sterilized by ultraviolet irradiation and immersed in the culture medium for 24 h). The control group and the blank group included cell wells cultured with culture medium and blank wells without culture medium, respectively, with 6 wells in each group. The entire 96-well plate was cultured in a cell culture incubator for 24 and 48 h, respectively. After the culture was completed, the old culture medium in the wells was removed, and 180 μL of culture medium and 20 μL of MTT reagent (concentration of 5 mg / mL) were added to each well after rinsing with sterile PBS, and the culture was continued for 4 hours. Then the medium was poured off, 200 μL of DMSO solvent was added to each well, and the wells were shaken in a shaker at 37°C and 120 rpm / min for 15 min, and then the absorbance at 570 nm was measured using an enzyme-linked microplate reader.

[0119] from Figure 6 It can be seen from the test results that the bio-based polyurethane elastomers prepared in Examples 1-5 and Comparative Examples 1-3 have good biocompatibility.

[0120] Degradation test: The bio-based polyurethanes prepared in Examples 1-5 and Comparative Examples 1-3 were tested for degradation performance. The specific method was as follows: 0.1 g of bio-based polyurethane was immersed in a PBS solution (pH = 7.4) containing 1 mg / mL porcine pancreatic lipase, placed in an oven at 37°C, and weighed every four weeks. The pH value was kept constant by renewing the PBS solution every week. The test process included rinsing the sample with distilled water, drying it in a vacuum oven at 40°C for 24 hours, and then weighing it. The degradation of bio-based polyurethane was observed at different times to determine the degradation ability of bio-based polyurethane.

[0121] The results are as follows Figure 7 As shown, from Figure 7 It can be seen from the test results that the bio-based polyurethane elastomers prepared in Examples 1-5 and Comparative Examples 1-3 all have certain degradation properties, among which Comparative Example 2 degrades the fastest, degrading 60% in 7 weeks, which indicates that the bio-based polyurethane of the present invention can be degraded in an enzyme environment.

[0122] Osteogenesis test: The bio-based polyurethanes prepared in Examples 1-5 and Comparative Examples 1-3 were subjected to an osteogenic test. Figure 8 and Fig. 9 They are the activity of the early osteoblast differentiation marker enzyme alkaline phosphatase (ALP) and the mineralization of the extracellular matrix after the cells were cultured on the material surface for 7 days and 14 days respectively. The larger the absorbance value, the higher the degree of mineralization of the cells. As can be seen from the results, the ALP activity of the cells on the surface of the material in Example 2 is the highest and the degree of mineralization is the highest. The above results prove that the synthesized bio-based polyurethane can not only stimulate osteoblast proliferation and osteogenic differentiation in the middle and late stages of implantation, but also promote extracellular matrix mineralization, i.e., bone formation.

[0123] The bio-based polyurethane of the Janus structure of the present invention has a hydrophilic chain and a hydrophobic chain in its molecular structure through ingenious molecular design, and forms a structure with one hydrophilic side and one hydrophobic side after water induction. The hydrophilic side organizes fibroblast migration and the hydrophobic side promotes bone regeneration. This special structure is adapted to the environmental requirements of periodontal tissue growth. Therefore, the present invention effectively overcomes some practical problems in the prior art and has high utilization value and use significance.

[0124] The above embodiments are merely illustrative of the principles and effects of the present invention, and are not intended to limit the present invention. Anyone familiar with the art may modify or alter the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or alterations made by a person of ordinary skill in the art without departing from the spirit and technical concept disclosed by the present invention shall still be covered by the claims of the present invention.

Claims

1. A Janus structure bio-based polyurethane membrane for inducing periodontal tissue regeneration, characterized in that: It is prepared by including the following main components, and the mass fraction of each component is: 100 parts of hydrophilic glycol; 100 parts of hydrophobic glycol; Isophorone diisocyanate 22.2-44.4 parts; Lysine diisocyanate 22.6-45.2 parts; 5.9-17.8 parts of chain extender E1 containing tertiary amine structure; 5.9-17.8 parts of chain extender E2 containing tertiary amine structure; 8.2-24.7 parts of monohalogenated alkanes; 3,4-dihydroxybenzoic acid 7.7-23.1 parts; 0.4 parts of organic bismuth catalyst; The chain extender E1 containing a tertiary amine structure is a compound including the following structural formula: (1); In formula (1), n=1-4; The chain extender E2 containing a tertiary amine structure is a compound including the following structural formula: (2); In formula (2), R is one of methyl, ethyl and isopropyl; The hydrophilic diol includes at least one of terminal hydroxyl polybutadiene and polyethylene glycol; The hydrophobic diol includes at least one of bio-based degradable polycaprolactone diol, polydimethylsiloxane diol, polytetramethylene glycol, polylactic acid diol and polypropylene glycol.

2. The Janus structure bio-based polyurethane membrane for inducing periodontal tissue regeneration according to claim 1, characterized in that: The preparation method comprises the following steps: The hydrophilic diol and the hydrophobic diol are dried at 110-130° C. and -0.1 MPa for 1-2 hours, and cooled to room temperature to obtain the dehydrated hydrophilic diol and the hydrophobic diol; The dehydrated hydrophilic diol and the hydrophobic diol are poured into a reactor, isophorone diisocyanate, lysine diisocyanate and 0.4 parts of an organic bismuth catalyst are added, and the mixture is stirred at 90-95° C. to be uniformly mixed and reacted for 2-3 hours to obtain a bio-based polyurethane prepolymer; The bio-based polyurethane prepolymer is cooled to 70-80° C., and a chain extender E2 containing a tertiary amine structure and a monohalogenated alkane are added to carry out chain extension and quaternization reaction for 2-3 hours to obtain a monofunctional bio-based polyurethane slurry; Adding chain extender E1 to the above monofunctional bio-based polyurethane slurry, reacting at 70-80° C. for 1-2 hours, cooling to 50-60° C., adding 3,4-dihydroxybenzoic acid and reacting for 1-2 hours to obtain a bifunctional bio-based polyurethane slurry; Finally, the above-mentioned bifunctional bio-based polyurethane slurry was cast into a polytetrafluoroethylene plate mold to form a film, immersed in deionized water, and placed in a 60-100°C oven for 24-48 hours to complete the migration process, thereby obtaining a Janus structure bio-based polyurethane film that induces periodontal tissue regeneration.

3. The Janus structure bio-based polyurethane membrane for inducing periodontal tissue regeneration according to claim 1, characterized in that: The monohalogenated alkane is a compound including the following structural formula: (3); In formula (3), m=4 or 6 or 8, and X is Br or I.

4. The Janus structure bio-based polyurethane membrane for inducing periodontal tissue regeneration according to claim 1, characterized in that: The number average molecular weight of the hydrophilic diol and the hydrophobic diol is M n =1000-4000 g / mol.

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