CN103656745B - Gel-poly(lactide-co-glycolide) (PLGA) two-phase gradient transition cartilage-bone repair material and preparation thereof - Google Patents
Gel-poly(lactide-co-glycolide) (PLGA) two-phase gradient transition cartilage-bone repair material and preparation thereof Download PDFInfo
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- CN103656745B CN103656745B CN201310673227.8A CN201310673227A CN103656745B CN 103656745 B CN103656745 B CN 103656745B CN 201310673227 A CN201310673227 A CN 201310673227A CN 103656745 B CN103656745 B CN 103656745B
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Abstract
The invention discloses a gel-poly(lactide-co-glycolide) (PLGA) two-phase gradient transition cartilage-bone repair material and preparation thereof. The material is cylindrical and consists of an upper material and a lower material, wherein the upper material is glycidyl methacrylate-modified carboxymethyl chitosan and gelatin; the lower material is porous PLGA. The preparation process comprises the following steps: preparing a gelatin microsphere serving as a pore-forming agent into a PLGA porous scaffold of which one end face is a conical concave surface; preparing a gel solution by using the modified chitosan and modified gelatin; dripping the gel solution on the PLGA porous scaffold of the conical concave surface, inverting, performing ultraviolet light cross-linking to obtain the gel-PLGA two-phase gradient transition cartilage-bone repair material. The material has high biocompatibility and biodegradability, is in a two-phase gradient transition structure, is tightly combined without drop and is simple in preparation process.
Description
Technical field
The present invention relates to a kind of gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating bracket material of (PLGA) two-phase gradient transition and preparation method, belong to cartilage-bone interface tissue repair technical field of biological material.
Background technology
It is inhomogenous that the bone interfaces such as cartilage-bone, tendon-bone, ligament-bone are organized on anatomical structure and component of organization, and possess the feature of structure and the change of composition continuous gradient, so the good reparation in order to realize Interface Microstructure, need support (the Leong K F of the functionalization gradient of design ideal, Chua C K, Sudarmadji N, Yeong W Y, Journal of the Mechanical Behavior of Biomedical Materials, 2008,1 (2): 140-152).
Be directed to cartilage-bone interface, it has significant difference in composition, structure and mechanical property, at present, by report prepared form, the gradient scaffolding of structure, bioactie agent content.The people such as Tempieri pass through induced biomineralization legal system for the hyaluronic acid magnesium-collagen composite support of composition and pattern Gradient distribution, the different region of this support can optionally inducing cell to skeletonization or cartilage differentiation (Tampieri A, Sandri M, Landi E, Pressato D, Francioli S, Quarto R, Martin I, Biomaterials, 2008,29 (26): 3539-3546).The people such as Kon prepare the shelf layer of different I Collagen Type VI and hydroxyapatite content respectively, then obtain forming three layers of different supports, clinical experiment gradient scaffolding being carried out to articular cartilage reparation shows that gradient scaffolding compares safety and stability, patient obtains comparatively satisfied therapeutic effect (Kon E, Delcogliano M, Filardo G, Pressato D, Busacca M, Grigolo B, Desando G, Marcacci M, Injury-International Journal of the Care of the Injured, 2010,41 (7): 778-786).In order to better simulate the mechanical property of cartilage-bone interface, and support cartilage and osteoblastic growth and differ entiation respectively, researcher has prepared the heterogeneous support of attached gel and porous support.It is agarose gel that the people such as Rodrigues prepare upper strata, lower floor is the two-phase support of starch/polycaprolactone, support that amniotic fluid stem cell is to cartilage and Osteoblast Differentiation (Rodrigues M T, Lee S J, Gomes M E respectively, Reis R L, Atala A, Yoo J J, Acta Biomaterialia, 2012,8 (7): 2795-2806).In two-phase support, gel phase can well simulate the tumor growth environment of chondrocyte, supports that stem cell breaks up to cartilage direction; And hard part can provide the mechanical property of suitable bone, promote that stem cell breaks up to bone.
For the gradient transition feature of cartilage-bone interface composition and character, need the induction of continuous gradient composition and biological activity signal to strengthen the reparation of cartilage-bone interface, therefore researcher proposes support (Mohan N, Dormer N H, the Caldwell K L that preparation has continuous gradient physics and chemical property, Key V H, Berkland C J, Detamore M S, Tissue Engineering Part A, 2011,17 (21/22): 2845-2855).At present, microfluidic device and electrospinning device is utilized can to have prepared electrospinning membrane support (Zhang X, Gao XH, Jiang L, Qin J H, Langmuir, 2012,28 (26), 10026-10032 of continuous gradient; Li XR, MacEwan M R, Xie J W, Siewe D, Yuan X Y, Xia Y N, Advance Functional Materials, 2010,20(10): 1632-1637).
The preparation of the existing heterogeneous gradient scaffolding for cartilage-Bone Defect Repari mostly can not realize the continuous gradient of transition region component, the separation and each interlayer easily comes off, adopt pyramidal structure to realize upper and lower two alternate continuous gradient transition, gel phase partial penetration makes biphase preparation method of combining closely have no bibliographical information to PLGA porous support simultaneously.
Summary of the invention
The object of the present invention is to provide a kind of gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of (PLGA) two-phase gradient transition and preparation method.This material has good bio-compatible and degradability, and material is biphase gradient transition structure, in conjunction with closely not coming off, and preparation process is simple.
For achieving the above object, the present invention is realized by following technical proposals: a kind of gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of (PLGA) two-phase gradient transition, it is characterized in that, this its shape of cartilage-bone renovating material is cylinder, be combined into by embedding with concave cone between upper and lower two kinds of different materials and two sections of material, wherein lower material to be aperture be 350 ~ 450 μm poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) material, top material is be 1:(8 ~ 10 in mass ratio by the gelatin of the carboxymethyl chitosan of glycidyl methacrylate modification and glycidyl methacrylate modification) form.
Above-mentioned poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) number-average molecular weight of (PLGA) is (5 ~ 12) ' 10
4; The molecular weight of the gelatin of glycidyl methacrylate modification is (1 ~ 7) ' 10
4; The weight average molecular weight of the carboxymethyl chitosan of glycidyl methacrylate modification is (0.8 ~ 1.2) ' 10
4.
Above-mentioned gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) preparation method of cartilage-bone renovating material of (PLGA) two-phase gradient transition, it is characterized in that comprising following process:
1) be that the gelatine microsphere of 350 ~ 450 μm is put in column-form by diameter, and make gelatine microsphere upper surface be formed into taper seat, be the solution of 8/2 to gelatin layer instillation ethanol with water ratio, at 37 DEG C, be cross-linked 10 ~ 30min, obtain gelatin skeleton;
2) gelatin skeleton obtained for step 1) is immersed mass concentration be 5 ~ 9% poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) dichloromethane solution in, make solution fully enter in gelatin gap, form gelatin cylinder.Gelatin cylinder is taken out, at-20 DEG C, carries out being separated 12h, replace dichloromethane solvent in ethanol, finally in the water of 37 DEG C, wash away gelatin, to obtain one end be the aperture of rounding cone mouth be 350 ~ 450 μm poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cylinder of (PLGA) material, it is deposited for subsequent use at-20 DEG C;
3) 1:(6 ~ 10 in mass ratio) by the gelatin of the carboxymethyl chitosan of glycidyl methacrylate modification and glycidyl methacrylate modification, at 40 DEG C, be dissolved in concentration be 0.1M, pH is in the phosphate buffer of 7.4, be mixed with the solution that mass concentration is 9%, adding with the molecular weight of 1 ~ 2 times of the gelatin quality sum of modified carboxy methyl chitosan and modification again in solution is the Polyethylene Glycol of 875, then add by modified carboxy methyl chitosan and modified gelatin gross mass 1 ~ 5% photocrosslinking agent 2959, obtain gel precursor solution;
4) by step 2) obtained porous poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cylinder of (PLGA) material puts into the pipe closed isodiametric one end upward, then in pipe, drips the obtained gel precursor solution of step 3) with rounding cone mouth, drip height and porous poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) cylinder is contour, is then inverted by this pipe at once, puts into UV-crosslinked case, is to irradiate crosslinked 5 ~ 10min under the ultraviolet light of 365nm at wavelength, obtain gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of (PLGA) two-phase gradient transition.
The invention has the advantages that, poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) realizes continuous gradient transition with the gel phase of modification of chitosan and modified gelatin with concave cone embedded structure, and two alternate not easily separated coming off of combining closely, its preparation process is simple.Meanwhile, this cartilage-bone renovating material has good biocompatibility, biodegradability, can be used for cartilage-bone interface tissue repair technical field of biological material.
Detailed description of the invention
embodiment 1
1. taking 0.11g diameter is that to be put in diameter be in the column template of 6mm for the gelatine microsphere of 350 ~ 450 μm, then (diameter is 5mm to the most advanced and sophisticated glass mold for taper of press-in, cone height is 2mm), instill 100 μ L ethanol and water ratio is the solution of 8/2, at 37 DEG C, crosslinked 15min, obtains gelatin skeleton.
2. gelatin skeleton is immersed concentration be 7% poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) dichloromethane solution in, soak 12h, in vacuum drying oven, pump gas in hole, ensure solution enter completely.Take out support, blot surface solution, in refrigerator, (-20 DEG C) are separated 12h, ethanol replacement 12h.Finally in the water of 37 DEG C, wash away gelatin skeleton, obtain highly for 4mm, diameter is 5mm, an end face be taper concave surface poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) porous support.Support is dry in vacuum drying oven, deposit for-20 DEG C.
3. take the carboxymethyl chitosan of 8.9mg glycidyl methacrylate modification and the gelatin of 71.1mg glycidyl methacrylate modification, at 40 DEG C, be dissolved in 1mL concentration is 0.1M, pH is in the phosphate buffer of 7.4, adding 80 μ L molecular weight is the Polyethylene Glycol of 875, then the concentration adding 16 μ L is the alcoholic solution of the photocrosslinking agent 2959 of 100mg/mL, obtains gel solution.
4. by obtained for step 2 poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) support is put in the glass tubing that the diameter closed one end is 5mm so that taper is concave upright, and then the gel solution dripped in glass tubing obtained by step 3, dripping is highly 4mm, rapidly glass tubing is inverted, then put it in UV-crosslinked case, crosslinked 7min under 365nm length ultraviolet light, obtain gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of (PLGA) two-phase gradient transition.
embodiment 2
1. taking 0.15g diameter is that to be put in diameter be in the column template of 6mm for the gelatine microsphere of 350 ~ 450 μm, then (diameter is 5mm to the most advanced and sophisticated glass mold for taper of press-in, cone height is 3mm), instill 100 μ L ethanol and water ratio is the solution of 8/2, at 37 DEG C, crosslinked 30min, obtains gelatin skeleton.
2. gelatin skeleton is immersed concentration be 6% poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) dichloromethane solution in, soak 12h, in vacuum drying oven, pump gas in hole, ensure solution enter completely.Take out support, blot surface solution, in refrigerator, (-20 DEG C) are separated 12h, ethanol replacement 12h.Finally in the water of 37 DEG C, wash away gelatin skeleton, obtain highly for 6mm, diameter is 5mm, an end face be taper concave surface poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) porous support.Support is dry in vacuum drying oven, deposit for-20 DEG C.
3. take the carboxymethyl chitosan of 5.6mg glycidyl methacrylate modification and the gelatin of 50mg glycidyl methacrylate modification, at 40 DEG C, be dissolved in 1mL concentration is 0.1M, pH is in the phosphate buffer of 7.4, adding 70 μ L molecular weight is the Polyethylene Glycol of 875, then the concentration adding 10 μ L is the alcoholic solution of the photocrosslinking agent 2959 of 100mg/mL, obtains gel solution.
4. by obtained for step 2 poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) support is put in the glass tubing that the diameter closed one end is 5mm so that taper is concave upright, and then the gel solution dripped in glass tubing obtained by step 3, dripping is highly 6mm, rapidly glass tubing is inverted, then put it in UV-crosslinked case, crosslinked 10min under 365nm length ultraviolet light, obtain gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of (PLGA) two-phase gradient transition.
embodiment 3:
1. taking 0.11g diameter is that to be put in diameter be in the column template of 6mm for the gelatine microsphere of 350 ~ 450 μm, then (diameter is 5mm to the most advanced and sophisticated glass mold for taper of press-in, cone height is 2mm), instill 100 μ L ethanol and water ratio is the solution of 8/2, at 37 DEG C, crosslinked 15min, obtains gelatin skeleton.
2. gelatin skeleton is immersed concentration be 9% poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) dichloromethane solution in, soak 12h, in vacuum drying oven, pump gas in hole, ensure solution enter completely.Take out support, blot surface solution, in refrigerator, (-20 DEG C) are separated 12h, ethanol replacement 12h.Finally in the water of 37 DEG C, wash away gelatin skeleton, obtain highly for 4mm, diameter is 5mm, an end face be taper concave surface poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) porous support.Support is dry in vacuum drying oven, deposit for-20 DEG C.
3. take the carboxymethyl chitosan of 12mg glycidyl methacrylate modification and the gelatin of 90mg glycidyl methacrylate modification, at 40 DEG C, be dissolved in 1mL concentration is 0.1M, pH is in the phosphate buffer of 7.4, adding 100 μ L molecular weight is the Polyethylene Glycol of 875, then the concentration adding 30 μ L is the alcoholic solution of the photocrosslinking agent 2959 of 100mg/mL, obtains gel solution.
4. by obtained for step 2 poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) (PLGA) support is put in the glass tubing that the diameter closed one end is 5mm so that taper is concave upright, and then the gel solution dripped in glass tubing obtained by step 3, dripping is highly 6mm, rapidly glass tubing is inverted, then put it in UV-crosslinked case, crosslinked 5min under 365nm length ultraviolet light, obtain gel-poly-(lactide-
co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of (PLGA) two-phase gradient transition.
Claims (3)
- A 1. gel-poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of two-phase gradient transition, it is characterized in that, this its shape of cartilage-bone renovating material is cylinder, be combined into by embedding with concave cone between upper and lower two kinds of different materials and two sections of material, wherein lower material to be aperture be 350 ~ 450 μm poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) material, top material is be 1:(8 ~ 10 in mass ratio by the gelatin of the carboxymethyl chitosan of glycidyl methacrylate modification and glycidyl methacrylate modification) form.
- 2. by gel according to claim 1-poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of two-phase gradient transition, it is characterized in that, poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) number-average molecular weight be (5 ~ 12) × 10 4; The molecular weight of the gelatin of glycidyl methacrylate modification is (1 ~ 7) × 10 4; The weight average molecular weight of the carboxymethyl chitosan of glycidyl methacrylate modification is (0.8 ~ 1.2) × 10 4.
- 3. one kind by gel according to claim 1-poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) preparation method of cartilage-bone renovating material of two-phase gradient transition, it is characterized in that comprising following process:1) be that the gelatine microsphere of 350 ~ 450 μm is put in column-form by diameter, and make gelatine microsphere upper surface form taper seat, be the solution of 8/2 to gelatin layer instillation ethanol with water ratio, at 37 DEG C, be cross-linked 10 ~ 30min, obtain gelatin skeleton;2) gelatin skeleton obtained for step 1) is immersed mass concentration be 5 ~ 9% poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) dichloromethane solution in, make solution fully enter in gelatin gap, form gelatin cylinder; Gelatin cylinder is taken out, at-20 DEG C, carries out being separated 12h, replace dichloromethane solvent in ethanol, finally in the water of 37 DEG C, wash away gelatin, to obtain one end be the aperture of rounding cone mouth be 350 ~ 450 μm poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) cylinder of material, it is deposited for subsequent use at-20 DEG C;3) 1:(8 ~ 10 in mass ratio) by the gelatin of the carboxymethyl chitosan of glycidyl methacrylate modification and glycidyl methacrylate modification, at 40 DEG C, be dissolved in concentration be 0.1M, pH is in the phosphate buffer of 7.4, be mixed with the solution that mass concentration is 9%, adding with the molecular weight of 1 ~ 2 times of the gelatin quality sum of modified carboxy methyl chitosan and modification again in solution is the Polyethylene Glycol of 875, then add by modified carboxy methyl chitosan and modified gelatin gross mass 1 ~ 5% photocrosslinking agent 2959, obtain gel precursor solution;4) by step 2) obtained porous poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) cylinder of material puts into the pipe closed isodiametric one end upward, then in pipe, drips the obtained gel precursor solution of step 3) with rounding cone mouth, drip height and porous poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) cylinder is contour, is then inverted by this pipe at once, puts into UV-crosslinked case, is to irradiate crosslinked 5 ~ 10min under the ultraviolet light of 365nm at wavelength, obtain gel-poly-(lactide- co-Acetic acid, hydroxy-, bimol. cyclic ester) cartilage-bone renovating material of two-phase gradient transition.
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| CN111363169A (en) * | 2017-09-03 | 2020-07-03 | 湖南七纬科技有限公司 | Preparation method of pH sensitive hydrogel |
| CN107715175B (en) * | 2017-09-30 | 2018-10-23 | 南京师范大学 | A kind of compound porous bone tissue engineering scaffold of PLGA/ amphoteric ions and preparation method thereof |
| CN111607047B (en) * | 2020-06-16 | 2021-03-26 | 浙江大学 | High-throughput biomaterial screening bidirectional gradient hydrogel and preparation method thereof |
| CN111607046B (en) * | 2020-06-16 | 2021-03-26 | 浙江大学 | Concentration/modulus bidirectional gradient hydrogel for high-throughput screening of biological materials and preparation method thereof |
| CN112516388B (en) * | 2020-12-01 | 2021-11-02 | 四川大学 | Domain-controllable osteochondral repair composite scaffold and preparation method thereof |
| CN116392639B (en) * | 2023-02-17 | 2024-02-13 | 无锡市中医医院 | Full-layer repair double-layer bracket and preparation method and application thereof |
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