阅读说明：本技术 一种液晶态复合纤维膜及其制备与应用 (Liquid crystal state composite fiber membrane and preparation and application thereof ) 是由 李娜 李霞 艾毅龙 黄达鸿 于 2021-07-23 设计创作，主要内容包括：本发明公开了一种液晶态复合纤维膜及其制备与应用。本发明方法包括如下步骤：胆固醇壬酸酯和胆固醇油烯基碳酸酯按照配方配比混合,加热至两种液晶化合物均到达清亮点温度以上,充分搅拌至完全混合,冷却得到粉末,获得CN-COC二元共混液晶,然后和聚氨酯弹性体按照配方配比加入到二氯甲烷中,搅拌混合物直到溶质完全溶解,通过静电纺丝技术获得复合纤维膜。该液晶态复合膜既有液晶态有序流动性和良好的生物学性能,同时又兼具聚氨酯的弹性特征,具有良好生物学性能和力学性能,可用于构建仿生命体细胞外基质的生物膜模型,并在此基础研究生命体液晶性对细胞行为的影响,从而为探明生命体微环境内细胞和基质相互作用提供理论和技术支撑。(The invention discloses a liquid crystal composite fiber membrane and preparation and application thereof. The method comprises the following steps: mixing cholesterol pelargonate and cholesterol oleyl carbonate according to a formula ratio, heating until two liquid crystal compounds reach a clearing point temperature, fully stirring until the two liquid crystal compounds are completely mixed, cooling to obtain powder, obtaining CN-COC binary blended liquid crystal, adding the CN-COC binary blended liquid crystal and polyurethane elastomer into dichloromethane according to the formula ratio, stirring the mixture until a solute is completely dissolved, and obtaining the composite fiber membrane by an electrostatic spinning technology. The liquid crystal state composite membrane has liquid crystal state ordered fluidity and good biological performance, has the elastic characteristic of polyurethane, has good biological performance and mechanical performance, can be used for constructing a biomembrane model imitating an extracellular matrix of a living body, and researches the influence of the crystallinity of the living body fluid on cell behaviors on the basis, thereby providing theoretical and technical support for exploring the interaction of cells and matrixes in the microenvironment of the living body.)

1. A liquid crystal composite fiber membrane characterized by:

the liquid crystal state composite fiber membrane is composed of a CN-COC binary blended liquid crystal formed by cholesterol pelargonate (CN) and Cholesterol Oleyl Carbonate (COC) and a polyurethane elastomer (PU); the content of the cholesterol pelargonate (CN) in the CN-COC binary blended liquid crystal is 20-50 wt%, and the mass fraction of the CN-COC binary blended liquid crystal in the liquid crystal composite fiber membrane is 30-50 wt%.

2. The liquid-crystalline composite fiber film according to claim 1, characterized in that:

the ratio of the cholesterol pelargonate (CN) in the CN-COC binary blended liquid crystal is 30 to 40 weight percent;

the mass fraction of the CN-COC binary blended liquid crystal in the liquid crystal composite film is 30-40 wt%.

3. The liquid-crystalline composite fiber film according to claim 1, characterized in that:

the polyurethane elastomer (PU) is polyether polyurethane; further, the polyurethane is medical polyether type EG-80A of Luborun, Germany.

4. The method for producing a liquid-crystal-state composite fiber film according to any one of claims 1 to 3, characterized in that: the method comprises the following steps:

(1) preparing CN-COC binary blended liquid crystal:

mixing cholesterol pelargonate (CN) and Cholesterol Oleyl Carbonate (COC) according to a formula ratio, heating until the two liquid crystal compounds reach a clearing point temperature or above, fully stirring until the two liquid crystal compounds are completely mixed, and cooling to obtain powder, namely obtaining the CN-COC binary blended Liquid Crystal (LC);

(2) preparing a liquid crystal state composite fiber membrane:

adding the CN-COC binary blend liquid crystal obtained in the step (1) into dichloromethane according to the formula ratio, stirring until the mixture is completely dissolved, adding a polyurethane elastomer (PU), continuously stirring the mixture until the solute is completely dissolved, and obtaining the liquid crystal state composite fiber membrane through electrostatic spinning.

5. The method for producing a liquid-crystal-state composite fiber film according to claim 4, characterized in that:

heating the two liquid crystal compounds in the step (1) until the two liquid crystal compounds reach the clearing point temperature, namely heating the two liquid crystal compounds to 95-100 ℃, and maintaining the two liquid crystal compounds for 3-5 min;

the amount of the dichloromethane in the step (2) is 6 to 10 percent of the mass fraction of the polyurethane elastomer in the system;

the electrostatic spinning conditions in the step (2) are as follows: the diameter of the rotating wheel is 10-15 cm, the rotating speed is 0-2000 rpm, the voltage is 10-20 KV, the injection speed is 1-2 mL/h, the receiving distance is 15-20 cm, the ambient temperature is 23-27 ℃, and the humidity is 70-80%.

6. The method for producing a liquid-crystal-state composite fiber film according to claim 5, characterized in that:

the amount of the dichloromethane in the step (2) is 8% of the mass fraction of the polyurethane elastomer in the system;

the electrostatic spinning conditions in the step (2) are as follows: the diameter of the rotating wheel is 13cm, the rotating speed is 1000rpm, the voltage is 15KV, the injection speed is 1.5mL/h, the receiving distance is 17.5cm, the ambient temperature is 25 ℃, and the humidity is 75%.

7. A preparation method of a mineralized liquid crystal state composite fiber membrane is characterized by comprising the following steps: comprising all the steps of the method for preparing the liquid crystal state composite fiber film according to any one of claims 4 to 6 and the following steps:

s1, preparing a mixed solution of calcium glycerophosphate and calcium chloride;

s2, soaking the liquid crystal state composite fiber membrane in the mixed solution obtained in the S1, adding alkaline phosphatase, mineralizing, washing, freezing and drying to obtain the mineralized liquid crystal state composite fiber membrane.

8. The method for preparing the mineralized liquid crystal state composite fiber membrane according to claim 7, wherein the method comprises the following steps:

in the step S1, the concentration of calcium glycerophosphate in the mixed solution is 90-100 mmol/L, and the concentration of calcium chloride is 60-70 mmol/L;

in the step S2, the addition amount of the alkaline phosphatase is calculated according to the concentration of the alkaline phosphatase in the mixed solution being 0.05-0.15 mg/L;

the mineralization time in the step S2 is 70-75 h.

9. A mineralized liquid crystal state composite fiber membrane is characterized in that: prepared by the preparation method of claim 7 or 8.

10. Use of the liquid crystal state composite fiber film according to any one of claims 1 to 3 or the mineralized liquid crystal state composite fiber film according to claim 9 for preparing a biomimetic biofilm.

Technical Field

The invention belongs to the field of tissue engineering, and particularly relates to a liquid crystal composite fiber membrane as well as preparation and application thereof.

Background

The liquid crystal state has both liquid fluidity and solid order, and is a substance state between a liquid state and a solid state. In biological systems, the combination of order and fluidity underlies the formation of tissues and structures, and therefore liquid crystals play a very important role in biology. Liquid crystal state structures exist in four compounds, namely lipid, protein, nucleic acid and polysaccharide which form a living body, and many functions of a living system cannot normally operate without the liquid crystal state structures. A large number of studies also show that liquid crystals have close relationship with biological structures, for example, sphingomyelin solution of nerve cells has the polarization characteristics of liquid crystals, muscle tissues and cells have molecular packing structures similar to liquid crystals, and photoreceptors (rods and cones) also have the characteristics of liquid crystal state substances. Therefore, the liquid crystal state biomaterial is constructed from the bionic angle, so that the novel bionic biomaterial can be developed, the microenvironment for in vivo tissue culture can be better simulated, a new thought and theoretical basis are provided for artificially simulating tissues and organs, and the method has important research significance.

Cholesterol is an important component of eukaryotic cell membranes and has an important function for maintaining the stability of the cell membranes, and derivatives of the cholesterol are liquid crystal materials with good biocompatibility, and particularly, the cholesterol ester liquid crystal is widely applied to the field of tissue engineering such as tissue engineering scaffolds, cell sensors and the like due to the good biocompatibility. Cholesterol Nonanoate (CN) and Cholesterol Oleyl Carbonate (COC) are two kinds of cholesterol ester liquid crystals with good biocompatibility, and researches show that special functional groups of the Cholesterol Nonanoate (CN) and the Cholesterol Oleyl Carbonate (COC) can trigger the secretion of IV type collagen, laminin, fibronectin and the like when contacting cells, and promote the adhesion and proliferation of the cells. The liquid crystal phase temperatures of CN and COC are 78.06-92.11 ℃ and 19.12-33.88 ℃ respectively, which is different from the physiological temperature of human body.

Polyurethane elastomer (PU) is a block copolymer consisting of hard crystalline segments dispersed between flexible amorphous segments. The hard segment is composed of aliphatic or aromatic isocyanates, promoting intermolecular hydrogen bonding and producing a glassy or crystalline phase that imparts toughness to the material. In contrast, soft segments are typically composed of polyethers, polybutadienes or polyesters, which are responsible for forming elastic domains, providing flexibility and elastic recovery to the polymer. In the current research, although PU has excellent mechanical properties, its biocompatibility is still far from that of ideal biomaterials.

Disclosure of Invention

To overcome the disadvantages and shortcomings of the prior art, the primary object of the present invention is to provide a method for preparing a liquid crystal composite fiber membrane.

Another object of the present invention is to provide a liquid crystalline state composite fiber film obtained by the above production method.

The invention further aims to provide application of the liquid crystal composite fiber membrane.

The purpose of the invention is realized by the following technical scheme:

a liquid crystal state composite fiber membrane is composed of a CN-COC binary blended liquid crystal composed of cholesterol pelargonate (CN) and Cholesterol Oleyl Carbonate (COC) and a polyurethane elastomer (PU); the content of the cholesterol pelargonate (CN) in the CN-COC binary blended liquid crystal is 20-50 wt%, and the mass fraction of the CN-COC binary blended liquid crystal in the liquid crystal composite fiber membrane is 30-50 wt%.

Further, the ratio of the cholesterol pelargonate (CN) in the CN-COC binary blended liquid crystal is 30-40 wt%; still further 30% by weight.

Further, the mass fraction of the CN-COC binary blended liquid crystal in the liquid crystal composite film is 30-40 wt%; still further 30% by weight.

Further, the polyurethane elastomer (PU) is a polyether urethane; further, the polyurethane is medical polyether type EG-80A of Luborun, Germany.

The preparation method of the liquid crystal state composite fiber membrane comprises the following steps:

(1) preparing CN-COC binary blended liquid crystal:

mixing cholesterol pelargonate (CN) and Cholesterol Oleyl Carbonate (COC) according to a formula ratio, heating until the two liquid crystal compounds reach a clearing point temperature or above, fully stirring until the two liquid crystal compounds are completely mixed, and cooling to obtain powder, namely obtaining the CN-COC binary blended Liquid Crystal (LC);

(2) preparing a liquid crystal state composite fiber membrane:

adding the CN-COC binary blend liquid crystal obtained in the step (1) into dichloromethane according to the formula ratio, stirring until the mixture is completely dissolved, adding a polyurethane elastomer (PU), continuously stirring the mixture until the solute is completely dissolved, and obtaining the liquid crystal state composite fiber membrane through electrostatic spinning.

Further, the clearing point temperature in the step (1) is as follows: cholesterol Nonanoate (CN)33.88 deg.C, Cholesterol Oleyl Carbonate (COC)92.11 deg.C.

Further, the heating in the step (1) until the two liquid crystal compounds reach the clearing point temperature is 95-100 ℃, and the heating is maintained for 3-5 min.

Further, the amount of the dichloromethane in the step (2) is 6 to 10 percent, and further 8 percent based on the mass fraction of the polyurethane elastomer in the system.

Further, the electrostatic spinning conditions in the step (2) are as follows: the diameter of the rotating wheel is 10-15 cm, the rotating speed is 0-2000 rpm, the voltage is 10-20 KV, the injection speed is 1-2 mL/h, the receiving distance is 15-20 cm, the ambient temperature is 23-27 ℃, and the humidity is 70-80%.

Further, the spinning technology in the step (2) is electrostatic spinning; further, the electrostatic spinning conditions are as follows: the diameter of the rotating wheel is 13cm, the rotating speed is 1000rpm, the voltage is 15KV, the injection speed is 1.5mL/h, the receiving distance is 17.5cm, the ambient temperature is 25 ℃, and the humidity is 75%.

A preparation method of a mineralized liquid crystal state composite fiber membrane comprises all the steps in the preparation method of the liquid crystal state composite fiber membrane and the following steps:

s1, preparing calcium glycerophosphate (Ca-GP) and calcium chloride (CaCl)₂) The mixed solution of (1);

s2, soaking the liquid crystal state composite fiber membrane in the mixed solution, adding alkaline phosphatase (ALP), mineralizing, washing, freezing and drying to obtain the mineralized liquid crystal state composite membrane.

Further, the concentration of calcium glycerophosphate in the mixed solution in the step S1 is 90-100 mmol/L, and the concentration of calcium chloride is 60-70 mmol/L; further, the concentration of calcium glycerophosphate was 95mmol/L and the concentration of calcium chloride was 64 mmol/L.

Further, in step S2, the amount of alkaline phosphatase added is calculated according to the concentration of the alkaline phosphatase in the mixed solution being 0.05-0.15 mg/L; further, it was calculated at 0.1 mg/L.

Further, the mineralization time in the step S2 is 70-75 h; further 72 h.

A mineralized liquid crystal state composite fiber membrane is prepared by the method.

The liquid crystal state composite fiber film or the mineralized liquid crystal state composite fiber film is applied to the preparation of the bionic biological film.

Furthermore, the bionic biomembrane is a guided bone regeneration biomembrane.

The principle of the invention is as follows: tecoflex EG-80A is degradable medical polyether polyurethane, and has good biocompatibility and elastic property. CN and COC are two kinds of micromolecular liquid crystals with good biocompatibility, and a CN-COC binary blending system which presents a liquid crystal state at physiological temperature can be obtained by controlling the proportion of the small molecule liquid crystals. The liquid crystal composite membrane prepared by combining the PU and the CN-COC blending system has the liquid crystal ordered fluidity, good biological performance and the elastic characteristic of PU, and is a composite material with good biological performance and mechanical performance. PU has certain compliance, so the PU fibre of receiving under static state presents the state of "noodle-like", and the fibre distributes disorderly and disorderly, gives the gyro wheel after accelerating to certain degree, and the gyro wheel fast turn can give the certain pulling force of fibre, leads to the fibre to present certain orientation, and LC PU who is spouted by the syringe needle also can receive certain shearing force effect simultaneously, can finally obtain the orderly oriented liquid crystal attitude ordered fiber membrane. In ALP/Ca-GP-CaCl₂Systematic oreCompared with the LC/PU fiber membrane in the liquid crystal state under the chemical conversion, the mineralization degree of the material is higher under the same mineralization time and the HAP crystals tend to be orderly arranged, which provides favorable conditions and material support for simulating the mineralization process of the micro-environment-liquid crystal state of the living body matrix.

Compared with the prior art, the invention has the following advantages and beneficial effects:

(1) CN and COC are cholesteric liquid crystals with good biocompatibility, which cannot take on a liquid crystal state at physiological temperature by their use alone due to the limitation of liquid crystal phase temperature. The molecular structure of the cholesterol derivative is cholest-5, 6-en-3 beta-R, CN and COC molecules are different from each other in substituent groups on 3 beta-C, CN molecules are substituted by a linear acyl chain consisting of 9 carbon atoms, COC molecules are substituted by an oleyl side chain consisting of 18 carbon atoms, unsaturated double bonds existing in the oleyl chain can form positive and negative isomers through bending deformation of the unsaturated double bonds, and the existence of the isomers can enlarge the transverse size among the molecules, so that the intermolecular force is reduced, and the melting temperature is lowered. While CN content increases (molecular size decreases), COC isomer decreases leading to a decrease in the intermolecular lateral dimension causing an increase in the phase transition temperature. Therefore, CN and COC are blended to form a CN-COC binary system, and the phase transition temperature can be adjusted by changing the proportion of CN and COC in the blending system. According to the invention, CN and COC are blended, and the liquid crystal phase temperature of a CN-COC blending system is regulated and controlled by utilizing the influence of the molecular configuration of the CN and COC, so that the cholesteric liquid crystal which is in a liquid crystal state at a physiological temperature is finally obtained, and the application research of the cholesteric small molecular liquid crystal in tissue engineering is facilitated.

(2) Although the CN-COC blending system with 20-50% of CN prepared by the invention has good biological and liquid crystal properties, the CN-COC blending system can not play a role of mechanical support when being used alone, so that the CN-COC blending system can be combined with PU with excellent elasticity for use, the liquid crystal property and the elasticity of the material can be comprehensively utilized, and the material with viscoelastic characteristics can be obtained, which is closer to the micro-environment mechanics in a living body.

(3) In the preparation process of the composite film material, an electrostatic spinning method is adopted, which is a very common method in extensive academic research, but the research on the application of the liquid crystal state fiber film to tissue engineering in particular is less common at present.

(4) The final purpose of the liquid crystal fibrous membrane prepared by the invention in the application aspect is to construct a biomembrane model imitating the extracellular matrix of a living body in vitro and study the influence of the crystallinity of the living body fluid on the cell behavior on the basis, thereby providing theoretical and technical support for exploring the interaction between cells and matrixes in the microenvironment of the living body.

(5) The liquid crystal state fiber membrane prepared by the invention has another purpose in the application aspect of discussing the action effect and the applicability of the liquid crystal state composite membrane as a bone tissue regeneration membrane and providing experiments and technical support for further exploring the applicability of liquid crystal state substances in the field of tissue engineering.

(6) The liquid crystal state fiber membrane prepared by the invention has another purpose in the application aspect of discussing the change of the material performance in the mineralization process of the bionic liquid crystal state composite membrane and the difference of the material mineralization and the non-liquid crystal state material mineralization.

(7) In the preparation of the CN-COC blending system, the CN content is selected to be 20-50 percent, mainly because the CN content is increased (the molecular size is reduced), the reduction of the COC isomer causes the reduction of the intermolecular transverse size to cause the phase transition temperature to be increased, and when the CN content is 20-50 percent, the physiological temperature is just in the liquid crystal phase temperature range.

(8) In the preparation of the CN-COC blending system, the most preferable CN content is 30 percent, mainly because when the CN content is 30 percent, the liquid crystal phase temperature range is 30.48-49.33 ℃, and the temperature range covers the physiological temperature optimally.

(9) In the preparation of the liquid crystal state composite film, the most preferable LC content is 30%, mainly because when the LC content is 30%, the liquid crystal phase temperature range of the composite film is 27.02-46.33 ℃, the liquid crystal phase temperature range is ideal, and the whole display material shows obvious liquid crystal property under polarized light.

(10) In the preparation of the liquid crystal state fiber membrane, an electrostatic spinning method is selected, and the method is simple and has strong operability.

(11) In the preparation of the liquid crystal fiber membrane, the total concentration of PU is selected to be 8%, the fiber fracture and poor mechanical property are easily caused by too low concentration, the solution is easily blocked and the uniformity of the fiber particle size is poor due to too high content, and therefore, the concentration of PU is selected to be 8%.

(12) The liquid crystal fibrous membrane prepared by the invention has good cell compatibility, and has the effects of promoting cell adhesion and proliferation compared with a pure PU membrane.

(13) The liquid crystal state fiber membrane prepared by the invention can simulate the ubiquitous structure state-liquid crystal state in a living body, and has an important effect on the deep research of the interaction between cells and materials in the living body.

(14) The method for regulating and controlling the mineralization of the liquid crystal material by utilizing the ALP has important significance for researching the mineralization process of tissues in a living body.

(15) The composite material has rich raw material sources, comprehensive functions and high market and research values.

Drawings

FIG. 1 is a DSC chart of series CN/COC blended liquid crystals (the curves correspond to CN-COC binary co-liquid crystals prepared by blending CN and COC in a mass ratio of 0:100, 100:0, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30 and 80:200 respectively from top to bottom).

FIG. 2 is a view showing a fiber film observed by a polarizing microscope; wherein A is PU fiber film, B is LC/PU composite fiber film.

FIG. 3 is a graph of cell proliferation.

FIG. 4 is a scanning electron micrograph of a preliminary cell experiment; wherein A is PU fiber film, B is LC/PU composite fiber film.

FIG. 5 shows SEM results of mineralization of a liquid crystal material; wherein A is an LC/PU composite fiber membrane, and B is an LC/PU composite fiber membrane mineralized for 72 hours.

FIG. 6 is a graph of Transwell cell crystal violet staining; wherein A is PU fiber film, B is LC/PU composite fiber film.

FIG. 7 is a graph of HE staining of a tissue section; wherein, A is the mineralized PU fiber film, and B is the mineralized LC/PU composite fiber film.

Detailed Description

The present invention will be described in further detail with reference to examples and drawings, but the embodiments of the present invention are not limited thereto. All the raw materials and reagents used in the present invention are commercially available conventional raw materials and reagents unless otherwise specified.

The PU used in the embodiment of the invention is biomedical polyether polyurethane EG-80A of Lubrizol company of Germany; cholesterol Nonanoate (CN) and Cholesterol Oleyl Carbonate (COC) were purchased from Sigma-Aldrich, USA.

EXAMPLE 1 preparation of LC/PU liquid crystalline composite fiber film

(1) Preparation of CN-COC binary blend Liquid Crystal (LC):

blending CN and COC according to the mass ratio of 0:100, 10:90, 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10 and 100:0 respectively. Adding the mixture into a glass tube, then placing the glass tube into an oven, adjusting the temperature to 95 ℃ and maintaining for 3min, wherein both liquid crystal compounds reach clearing points, then fully stirring until the two liquid crystals are completely mixed, cooling to obtain a powder sample, and placing the sample in the oven for storage for later use.

(2) Preparing a liquid crystal state composite fiber membrane:

accurately weighing CN-COC binary Liquid Crystals (LC) prepared by CN and COC in the step (1) according to the mass ratio of 30:70, adding the CN-COC binary liquid crystals into dichloromethane, and stirring at the rotating speed of 100r/min until the CN-COC binary liquid crystals are completely dissolved to obtain an LC/dichloromethane solution; accurately weighing PU according to the mass ratio of LC to PU of 10:90, 20:80, 30:70, 40:60 and 50:50 respectively, adding the PU into the obtained LC/dichloromethane solution (wherein the total concentration of PU is 2%, 4%, 6%, 8% and 10% respectively), and continuing stirring the mixture until the solute is completely dissolved to obtain an LC/PU mixed solution.

Introducing the completely dissolved LC/PU mixed solution into a micro-injection pump, selecting a rotating wheel with the diameter of 13cm, the rotating speed of 1000rpm, the voltage of 15KV, the injection speed of 1.5mL/h and the receiving distance of 17.5cm, and carrying out electrostatic spinning (the rotating wheel with the diameter of 13cm, the rotating speed of 1000rpm, the voltage of 15KV, the injection speed of 1.5mL/h, the receiving distance of 17.5cm, the ambient temperature of 25 ℃ and the humidity of 75%) at the ambient temperature of 25 ℃ and under the humidity of 75% to obtain 5 LC/PU composite fiber membranes.

The DSC chart of a series of CN-COC binary blended liquid crystals prepared in the example is shown in figure 1. Wherein, the temperature ranges of partial liquid crystal phases are respectively as follows: 19.12-33.88 deg.C (0:100), 22.14-39.12 deg.C (10:90), 24.61-42.25 deg.C (20:80), 30.48-49.33 deg.C (30:70), 32.15-53.053 deg.C (40:60), 35.42-57.27 deg.C (50: 50).

The liquid crystal phase temperature of the 5 CN-COC/PU fiber membranes prepared in the embodiment is increased along with the increase of the LC content, and finally, the LC/PU composite membrane prepared according to the LC-PU mass ratio of 30:70 and having the liquid crystal phase temperature range of 27.02-46.33 ℃ is selected as a final preparation sample, and subsequent performance analysis is carried out.

Example 2 analysis of Properties of LC/PU liquid-crystalline composite fiber film

Weighing about 6-10 mg of dried CN-COC and LC/PU composite membrane, sealing an aluminum crucible, putting the sealed aluminum crucible into a differential scanning calorimeter, heating the aluminum crucible to 100 ℃ from-20 ℃ at the speed of 5 ℃/min, keeping the temperature for 5min, cooling the aluminum crucible to-20 ℃, heating the aluminum crucible again, and then cooling the aluminum crucible to room temperature, wherein the whole test process is carried out at N₂Under protection. And observing and recording the temperature of the liquid crystal composite film in the change process, taking a picture, moving the liquid crystal phase temperature interval of the CN-COC blended liquid crystal to a high position along with the increase of the CN content, and more remarkably liquid crystallinity of the mixed film and moving the liquid crystal phase interval to the high position along with the increase of the LC content in the LC/PU mixed film.

Placing the CN-COC and LC/PU composite film on a glass sheet in a polarizing microscope, heating to 100 ℃ from-20 ℃ at a heating rate of 1 ℃/min by using LNP95, and cooling the sample by using liquid nitrogen at a speed of 1 ℃/min. The sample was then heated again and the state of the sample under a polarizing microscope was examined. The results are shown in FIG. 2. The results show that the LC property of the LC/PU composite film is more and more obvious along with the increase of the LC content, and the temperature interval of the liquid crystal phase is shifted to a higher position, which is consistent with the DSC test results.

③ placing the LC/PU composite membrane in a 24-hole plate, carrying out Co-60 Gamma (15 kGy) radiation sterilization for 6h, and then carrying out 75% (v/v) ethanol sterilization for 30 min. Mouse preadipocytes (MC3T3-E1) were seeded onto 24-well plates. Cells were allowed to adhere for 3h and sufficient media necessary was added to each well. Observing and analyzing the adhesion, proliferation and morphology of the cells after culturing for 1, 3, 5 and 7 days respectively. The results are shown in FIGS. 3 and 4. The results show that the LC/PU composite membrane prepared by the invention has better cell adhesion, proliferation and growth conditions than PU membranes, and the combined application of LC/PU can greatly improve the biological performance of PU.

Example 3 preparation of biomimetic mineralized LC/PU liquid crystalline composite fibrous Membrane

Blending CN and COC according to the mass ratio of 30: 70. The mixture was added to a glass tube, which was then placed in an oven, the temperature was adjusted to 95 ℃ and maintained for 3min, at which point both liquid crystal compounds reached a clearing point, followed by stirring thoroughly until both liquid crystals were completely mixed, and cooled to a powder sample. Then adding the mixture into dichloromethane, and stirring the mixture at the rotating speed of 100r/min until the mixture is completely dissolved to obtain an LC/dichloromethane solution; accurately weighing PU according to the mass ratio of LC to PU of 40:60, adding the PU into the obtained LC/dichloromethane solution (wherein the total concentration of PU is 8%), and continuing stirring the mixture until the solute is completely dissolved to obtain an LC/PU mixed solution.

And (3) introducing the completely dissolved LC/PU mixed solution into a micro-injection pump, selecting a roller with the diameter of 13cm, the rotating speed of 1000rpm, the voltage of 15KV, the injection speed of 1.5mL/h, the receiving distance of 17.5cm, and carrying out electrostatic spinning at the ambient temperature of 25 ℃ and the humidity of 75% to obtain the LC/PU composite fiber membrane.

Respectively soaking LC/PU composite fiber membrane and PU fiber membrane in Ca-GP (calcium glycerophosphate) and CaCl₂(calcium chloride) mixed solution (Ca: P1.67, Ca-GP concentration 95mmol/L, CaCl₂64mmol/L), a certain amount of ALP (alkaline phosphatase) is added to make the concentration of the ALP 0.1mg/L, the stent is taken out after mineralization for 72h, washed three times by deionized water and freeze-dried for 24h, and the mineralized film of the liquid crystal material and the non-liquid crystal material is obtained.

Example 4 modulation of cell behavior by ordered mobility of liquid crystals

In the field of tissue engineering, liquid crystal materials have been a focus of much attention, and especially cholesteric liquid crystals are regarded as important because of their particular cholesteric liquid crystal structure. However, the previous research on cholesteric liquid crystal is focused on the liquid crystallinity of the material, and actually, the research is focused on the substances which are in a liquid crystal state at physiological temperature in the field of tissue engineering. The CN-COC/PU fiber membrane material prepared in the embodiment can be in a liquid crystal state at a physiological temperature, and is also a theoretical technical support for researching the interaction between cells and matrix materials in a physiological environment.

The pre-embryonic osteoblast MC3T3-E1(ATCC) was cultured on PU and LC/PU for 5min, 1h, 2h, 4h, 8h, 12h and 24h, and then statistical analysis of the cell spreading area journality was carried out by AFM imaging technology. In addition, a single cell force spectrum technology is adopted, MC3T3-E1 is adhered to a functionalized AFM probe cantilever without a sharp point, the tensile breaking force between cells and a substrate material is measured in a ScanAsyst mode, and the influence of a liquid crystal microenvironment on cell adhesion and skeleton tension is researched by combining the measurement result of the Young modulus of the material. CCK8, cytoskeletal staining and Tran swell chamber analysis are carried out to analyze the influence of liquid crystal microenvironment on cell adhesion, migration, proliferation and activity, osteogenic differentiation conditions of stem cells are detected through osteogenic related genes such as OPN, Runx2 and COL-I, Affymetrix GeneChip is adopted to carry out Gene chip detection, and Collection analysis System V3.0(CapitalBio) is adopted to carry out Gene integration analysis, and Western blot is adopted to detect RhoA/ROCK channels. The Tran swell cell migration crystal violet staining results are shown in FIG. 6 and show: the cell number of the LC/PU group passing through the chamber is obviously higher than that of the PU group, and the cells rapidly proliferate and spread on the bottom of the whole chamber, which shows that the LC/PU material has more excellent cell compatibility than PU.

Example 5 osteopromotion Properties of biomimetic mineralized liquid crystalline Material

Mineralized products of the liquid crystalline material were obtained as in example 3, and SEM, XRD, FITR analyses were performed on the mineralized material to discuss the effect of the mineralization process on liquid crystalline and non-liquid crystalline materials, respectively. SEM results As shown in FIG. 5, the HAP layer adhered to the surface of the fibers was visible after 72h of mineralization.

After skin, subcutaneous tissues and periosteum are incised in the middle of the head of an SD rat, a full-layer skull defect model with the diameter of 5mm is constructed by blunt separation and exposed bone surfaces, mineralized products of liquid crystal state materials and non-liquid crystal state materials are filled in the bone defect part respectively, and then the periosteum, the subcutaneous tissues and the skin are sutured in a layered alignment mode. And analyzing the bone promoting performance of different membrane materials by micro-CT and tissue section staining respectively after 4, 8 and 12 weeks of operation. HE staining results are shown in fig. 7. The results show that: the growth condition of the new bone of the mineralized liquid crystal material is obviously better than that of the non-liquid crystal material.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.