CN103834041B

CN103834041B - MGF-Ct24E (Mechano-growth factor-Ct24E) functionalized polylactic acid bionic bone matrix material and preparation method thereof

Info

Publication number: CN103834041B
Application number: CN201410059996.3A
Authority: CN
Inventors: 彭坤
Original assignee: Chongqing Medical and Pharmaceutical College
Current assignee: Chongqing Medical and Pharmaceutical College
Priority date: 2014-02-21
Filing date: 2014-02-21
Publication date: 2017-02-08
Anticipated expiration: 2034-02-21
Also published as: CN103834041A

Abstract

The invention discloses an MGF-Ct24E (Mechano-Growth factor-Ct24E) functionalized polylactic acid bionic bone matrix material and a preparation method thereof. The preparation method comprises the following steps: using surface hydroxyl groups of hydroxyapatite (HAP) as triggering groups, grafting lactide onto the surface of the HAP by using a ring-opening polymerization method in a covalence chemical bond manner, and then introducing maleic anhydride with good cytocompatibility and the MGF-Ct24E with good bone inductivity, thus obtaining the MGF-Ct24E functionalized polylactic acid bionic bone matrix material. The bone regeneration and repair function of the MGF-Ct24E functionalized polylactic acid bionic bone matrix material can be promoted by introducing the MGF-Ct24E. The MGF-Ct24E functionalized polylactic acid bionic bone matrix material has good biocompatibility and can be directly applied to tissue repair. The preparation method is simple, practicable and good in repeatability.

Description

A kind of MGF-Ct24E Polylatides bionic bone matrix material and preparation method thereof

Technical field

The present invention relates to a kind of bone tissue engineering stent material and preparation method thereof, particularly a kind of MGF-Ct24E function Change polylactic acid bionic bone matrix material and preparation method thereof.

Background technology

Polylactic acid (polylactic acid, PLA) has good biocompatibility, biodegradability and its degraded The advantages of product can participate in internal tricarboxylic acid cycle, is widely used in operation suture thread, bone anchor, drug delivery system and group Knit the aspects such as engineering scaffold material.However, polylactic acid hydrophobicity is relatively strong, the acid accumulation in degradation process easily causes swelling and inflammation Disease is reacted, and lacks the signal identifying further, mechanical performance with active group and the cell-specific of bioactie agent reaction Poor the shortcomings of, limit its development in bone tissue engineer field.

Hydroxyapatite is the main component of skeleton, introduces hydroxyapatite and modifies PLA, can strengthen the machine of polymer Tool intensity and biological activity.Good cytocompatibility is had as bone tissue engineering stent material using maleic anhydride modified PLA Property, biological degradability and plasticity, but still lack the signal site of Osteoblast Specific identification, do not possess osteoinductive.

Mechano growth factor E domain 24 peptide（Tyr-Gln-Pro-Pro-Ser-Thr-Asn-Lys-Asn-Thr-Lys- Ser-Gln-Arg-Arg-Lys-Gly-Ser-Thr-Phe-Glu-Glu-His-Lys,MGF-Ct24E）Skeletal muscle can be promoted thin The propagation of born of the same parents and differentiation, thus promote bone injury reparation.

Content of the invention

The purpose of the present invention is to disclose a kind of to have good biodegradability, biocompatibility, mechanical strength and bone The new polylactic acid bionic bone matrix material of induced activity.

Employed technical scheme comprise that such for realizing the object of the invention, a kind of MGF-Ct24E Polylatides are imitated Osteogenic stromal material, its structural formula is：

What deserves to be explained is, above-mentioned MGF-Ct24E Polylatides bionic bone matrix material is with hydroxyapatite （Hydroxyapatite,HAP）, poly (l-lactic acid)（P oly L-la ctic acid,PLA）, maleic anhydride（Maleic anhydride,MA）And Mechano growth factor（Mechno-growth factor Ct24E,MGF-Ct24E）For raw material.With HAP table Face hydroxyl is initiating group, by ring-opening polymerisation method by lactide（Lactide,LA）It is grafted in the way of covalent chemical bond HAP surface, obtains nano level bone frame material PLA-HAP.It is re-introduced into that there is good cell compatibility MA, obtain active group MPLA-g-HAP.It is finally introducing the MGF-Ct24E with good osteoinductive, obtain MGF-Ct24E-MPLA-g-HAP Bionics Bone Host material.This material is optimized to the performance of polylactic acid by introducing HAP, MA and MGF-Ct24, has good biology Degradability, biocompatibility, mechanical strength and bone-inducting active.

Another purpose of this patent is that open one kind prepares above-mentioned MGF-Ct24E Polylatides bionic bone matrix material The method of material is it is characterised in that comprise the following steps：

1）Obtain dry lactide standby；Meanwhile, stannous octoate is dissolved in organic solvent, being made into concentration is 0.01% Stannous octoate solution；

2）Lactide and hydroxyapatite are pressed 9:1 mass ratio adds in reactor, then adds in described reactor Described stannous octoate solution；

3）After material in described reactor is uniformly mixed, then by described reactor evacuation；

4）Described reactor is placed in 140 DEG C of oil baths, after reaction 8～24h, the material purification in reactor obtains PLA-g-HAP；

5）By step 4）Gained PLA-g-HAP is dissolved in organic solvent, adds maleic anhydride and dibenzoyl peroxide, It is sufficiently stirred for, is dried under vacuum to constant weight at room temperature, obtain mixture；

6）Under the protection of nitrogen, by step 5）The mixture of gained is heated to molten condition, and stirring is not less than 6h, product Through after purification, obtaining MPLA-g-HAP；

7）By step 6）Gained MPLA-g-HAP is dissolved in dimethyl sulphoxide solution, stirs, and obtains MPLA-g- HAP solution；

8）Compound concentration is 1- (3- the dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride solution of 20mg/mL and dense Spend the N- hydroxy thiosuccinimide solution for 10mg/mL；Above two solution is added to step 7）The MPLA- being obtained In g-HAP solution, stirring is not less than 2h, obtains reactant mixture；

9）Mechano growth factor MGF-Ct24E is added step 8）In the reactant mixture being obtained, continue stirring and be not less than 2h, the product that obtains through washing, after purification, obtain product MGF-Ct24E-MPLA-g-HAP.

In said method, MGF-Ct24E Polylatides bionic bone matrix material is with hydroxyapatite（HAP）Surface Hydroxyl be initiating group, by ring-opening polymerisation method by lactide with covalent chemical bond by way of be grafted to HAP surface, obtain PLA- g-HAP.It is re-introduced into the maleic anhydride with good cell compatibility and the Mechano growth factor with good osteoinductive（MGF- Ct24E）.It is critical only that of method, in the way of covalent bond, polymer should be grafted to the more inorganic matters of content in osseous tissue The surface of composition HAP.Key is more to introduce Mechano growth factor（MGF-Ct24E）Bone tissue restoration can be promoted.

The solution have the advantages that mathematical.Experimental data shows, new material disclosed by the invention has relatively Strong mechanical property, possesses three-dimensional intercommunication cavernous structure and self-bone grafting biological activity, can be applied as bionic bone matrix material. Specifically, in MGF-Ct24E-MPLA-g-HAP bionic bone matrix material, each group is divided into by covalently bonded organizational project material Material, has following features：The percent grafting of MGF-Ct24E is 22.63%, and glass transition temperature is 58.62 DEG C, and elastic modelling quantity is 2.9 ± 1.7GPa, tensile strength are 65 ± 0.2Gpa, and fracture elongation is 7.2 ± 0.7%, and its mechanical property is substantially better than blending Section bar material, and material is in intercommunity cavernous structure.

Brief description

Fig. 1 is MGF-Ct24E Polylatides bionic bone matrix material（MGF-Ct24E-MPLA-g-HAP）Synthesis Route and structural representation；

Fig. 2 is MGF-Ct24E Polylatides bionic bone matrix material（MGF-Ct24E-MPLA-g-HAP）Infrared Absorption spectrum；

Fig. 3 is MGF-Ct24E Polylatides bionic bone matrix material（MGF-Ct24E-MPLA-g-HAP）DSC bent Line chart；

Fig. 4 is MGF-Ct24E Polylatides bionic bone matrix material（MGF-Ct24E-MPLA-g-HAP）Stretching Load-deformation curve；

Fig. 5 is MGF-Ct24E Polylatides bionic bone matrix material（MGF-Ct24E-MPLA-g-HAP）Scanning Electron microscope.

Specific embodiment

The invention will be further described with reference to the accompanying drawings and examples, but should not be construed the above-mentioned theme of the present invention Scope is only limitted to following embodiments.Without departing from the idea case in the present invention described above, known according to ordinary skill Know and customary means, make various replacements and change, all should include within the scope of the present invention.

According to MGF-Ct24E Polylatides bionic bone matrix material shown in Fig. 1, can be prepared（MGF-Ct24E-MPLA- g-HAP）.

First, thick LA100g ethyl acetate is washed three times, be vacuum dried 48h, standby.

Octoate catalyst stannous 0.001g are dissolved in 10ml dichloromethane, are made into the solution that concentration is 0.01%, standby.

LA and 2g n-HAP after taking 18g to process loads round-bottomed flask, adds the stannous octoate that concentration is 0.01% molten Liquid, after uniformly mixing, seals bottleneck, evacuation is to remove the air in bottle and dichloromethane.

Keep vacuum in reaction bulb, the round-bottomed flask that will be equipped with reactant is put in the thermostatical oil bath that temperature is 140 DEG C, After lactide is completely melt, start to clock, react 20h.

After question response terminates, product is removed reaction bulb, cooling.With chloroform dissolving, body is co-precipitated with chloroform/ethanol System, extracts product PLA-g-HAP；After preliminarily dried, dissolving purification again, repeats 2～3 times；Final product is in normal temperature condition Under, it is vacuum dried 48h, standby.

Take 10g PLA-g-HAP to be dissolved in CH2Cl2, add dibenzoyl peroxide and 1g MA, be sufficiently stirred for, in room temperature Under be dried under vacuum to constant weight, under nitrogen protective condition, using fusion method continuously stirred reaction 6h；By product CHCl3 and mistake Amount methanol purified product, obtains MPLA-g-HAP.

1g MPLA-g-HAP is added 25ml beaker and is dissolved in appropriate DMSO, at room temperature, magnetic agitation makes it dissolve Fully.Meanwhile, prepare 20mg/mL1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (N- (3- Dimethylamino propyl)-N-eth ylcarbodiimide hydrochloride crystalline, EDC) and 10mg/mL N- hydroxy thiosuccinimide (N-Hydroxy sulfosuccinimide sodium salt, sulfo- NHS solution for standby).

NHS and EDC solution are added in MPLA-g-HAP solution, preferably, three's volume ratio is：1:1:6, room temperature Lower continuation magnetic agitation reaction 2h.Mechano growth factor MGF-Ct24E is added in above-mentioned reactant mixture, magnetic agitation under room temperature Reaction 2h.Solution complete for above-mentioned reaction is instilled in excessive distilled water, collects the precipitate in distilled water, and precipitate is steamed Distilled water is washed to nonirritant taste, is placed in freeze drying box, is dried to constant weight, then proceedes to separate using THF-H2O Purification system purifies 3 times, to remove unreacted Mechano growth factor MGF-Ct24E and excessive NHS and EDC, that is, obtains MGF- Ct24E Polylatides bionic bone matrix material（MGF-Ct24E-MPLA-g-HAP）.

The infrared absorption spectroscopy of the MGF-Ct24E-MPLA-g-HAP that the present embodiment is obtained is as shown in Figure 2.

The DSC curve figure of the MGF-Ct24E-MPLA-g-HAP that the present embodiment is obtained is as shown in Figure 3.

The tensile stress-strain curve of the MGF-Ct24E-MPLA-g-HAP that the present embodiment is obtained is as shown in Figure 4.

The scanning electron microscope (SEM) photograph of the MGF-Ct24E-MPLA-g-HAP that the present embodiment is obtained is as shown in Figure 5.

The content situation of MGF-Ct24E such as table 1 institute in the MGF-Ct24E-MPLA-g-HAP molecule that the present embodiment is obtained Show：

Table 1

Claims

1. a kind of method preparing MGF-Ct24E Polylatides bionic bone matrix material is it is characterised in that include following walking Suddenly：

1) obtain dry lactide standby；Meanwhile, stannous octoate is dissolved in organic solvent, be made into concentration be 0.01% pungent Sour stannous solution；

2) lactide and hydroxyapatite are pressed 9:1 mass ratio adds in reactor, then adds described in described reactor Stannous octoate solution；

3) after the material in described reactor uniformly being mixed, then by described reactor evacuation；

4) described reactor is placed in 140 DEG C of oil baths, after reaction 6～24h, the material purification in reactor obtains PLA- g-HAP；

5) by step 4) gained PLA-g-HAP is dissolved in organic solvent, adds maleic anhydride and dibenzoyl peroxide, fully Stirring, is dried under vacuum to constant weight at room temperature, obtains mixture；

6) under the protection of nitrogen, by step 5) mixture of gained is heated to molten condition, and stirring is not less than 6h, and product passes through After purification, obtain MPLA-g-HAP；

7) by step 6) gained MPLA-g-HAP is dissolved in dimethyl sulphoxide solution, stirs, and obtains MPLA-g-HAP molten Liquid；

8) compound concentration is 1- (3- the dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride solution of 20mg/mL and concentration is The N- hydroxy thiosuccinimide solution of 10mg/mL；Above two solution is added to step 7) MPLA-g- that obtained In HAP solution, stirring is not less than 2h, obtains reactant mixture；

9) Mechano growth factor MGF-Ct24E is added step 8) in the reactant mixture that obtained, continue stirring and be not less than 2h, obtain The product that arrives through washing, after purification, obtain product MGF-Ct24E-MPLA-g-HAP.

2. the MGF-Ct24E Polylatides bionic bone matrix material that method as claimed in claim 1 obtains.