CN1429628A - Biodegradable composite material for promoting regeneration of blood vessel and its manufacturing method - Google Patents
Biodegradable composite material for promoting regeneration of blood vessel and its manufacturing method Download PDFInfo
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- CN1429628A CN1429628A CN 01145295 CN01145295A CN1429628A CN 1429628 A CN1429628 A CN 1429628A CN 01145295 CN01145295 CN 01145295 CN 01145295 A CN01145295 A CN 01145295A CN 1429628 A CN1429628 A CN 1429628A
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Abstract
A biodegradable copmosite material able to promote regeneration of blood vessel is prepared from a biodegradable material and phosphatide dispersed in the said biodegradable material in the micro-lipochondria mode. The derivative of phosphatide has high dispersity and chemo-attraction function, so it can cooperate with growth factor to promote the regeneration of blood vessel.
Description
Technical field
The present invention relates to a kind of revascularization guidance system, and be particularly related to a kind of Biodegradable composite material and manufacture method thereof that promotes revascularization.
Background technology
Blood vessel spreads all over organ and the tissue that the partes corporis humani divides, one of the organ of wanting for body weight for humans, and its topmost function is the conveying of blood, and human body is the circulation by blood, keeps that each organ and tissue can normal operation and metabolism in the body.Therefore, when human tissue organ in pathological changes or when damage, the degree of its revascularization usually is the important indicator that histoorgan is repaired.But some patient's pathological changes clinically is because health itself can't carry out revascularization normally, for example wound of chronic disease (as diabetic wounds, venous disease), coronary occlusion etc., and make wound can not be restored or normal operation.Therefore as how external or intravital method prepares or catalysis guides the formation of suitable regeneration vessel, will be a quantum jump of following medical skill, particularly field of tissue engineering technology, the also extensible new-type treatment that is applied to numerous disease of the correlation technique that is developed.
Revascularization must be via a succession of quite complicated program in the body, comprising the effect between cell and cell, the influence of somatomedin and the effect of extracellular matrix, injured or organizing of pathological changes at first can discharge regeneration vessel somatomedin (angiogenic growth factors) around injured tissue, and the somatomedin of these regeneration vessels is with after receptor (receptor) on the vascular endothelial cell (endothelial cells) combines, just can activating cell and secrete some specific enzyme and somatomedin, and these specific enzymes and somatomedin can decompose the basement membrane of original blood vessel, make vascular endothelial cell can move to impaired tissue, produce the mechanism of vascular repair or regeneration vessel.The regulation and control of this revascularization or inhibition have represented surprising potentiality on treatment of diseases.In the regenerative process of blood vessel, the somatomedin of regeneration vessel has an important adjusting function, therefore, the research group (comprising university and many large and small companies) that many tips are arranged in recent years, actively drop into great amount of manpower and material resources, look for suitable regeneration vessel somatomedin and medicine, attempt carrying out the preparation of regeneration vessel in the vitro tissue cultured method.In addition, also there is the research group to attempt the regeneration vessel somatomedin is added on the wound dressing, promotes the healing of wound.
The exploitation organic regeneration technology, blood vessel is indispensable part.Obtain a large amount of blood inflows in order to make the organizational project gained cultivate organ (as pancreas, liver, liver, kidney), promote that blood vessel hyperplasia (angiogenesis) is a key, the suitable revascularization material of framework how, making revascularization be quickened also to carry out smoothly, is the important goal of tissue engineering technique.Being usually used in promoting the material of revascularization at present, is that substrate attaches cell with chemical high molecular polymer usually, and adds the somatomedin promotion cell growth that promotes revascularization.Problems such as with chemical high molecular polymer is that the substrate palpus is considered biocompatibility etc., adds to promote the somatomedin of revascularization then must consider molecular diffusion, and protein denaturation is lost activity, and preservation is difficult and cost an arm and a leg.Therefore develop high-biocompatibility, low cost, high efficiency natural biological polymer composite will help to promote the carrying out of this technology.
More existing basic results of study, typically using somatomedin stimulates role of Vasculogenesis, though the division that somatomedin can irritation cell promotes moving and forming tubular structure of cell, following defective is arranged but:
1. molecular weight is excessive, slow down the speed of diffusion: its molecular weight of general somatomedin (for example: bFGF and VEGF) is distributed between 1～20,000 mostly, so big molecular weight can't effectively spread in the matter between tissue, is difficult for forming the Concentraton gradient of a somatomedin.Because this defective makes its effectively direction of cell guiding growth;
2. may become the target of immune system attack: somatomedin may bring out immunoreation in heterologous organism, and is light then just slacken the effect of somatomedin, heavy then cause anaphylaxis to cause organism death;
3. changeableness: most somatomedin all is to be made of protein because its structure and effect are complicated, so when physics or chemical environment change (for example: the process of processing) degeneration takes place easily, cause active forfeiture;
4. price is high: somatomedin is that the mode manufacturing by gene recombinaton forms mostly, so price is high.
Summary of the invention
In view of this, primary and foremost purpose of the present invention is to provide a kind of Biodegradable composite material that promotes revascularization.
Another purpose of the present invention is to provide a kind of method for making that promotes the biodegradable composite material of revascularization.
For reaching above-mentioned purpose, revascularization guidance system provided by the present invention is one to comprise the biodegradability base material of phospholipid, can have the usefulness of effective enhancement revascularization.Because the derivant of phospholipid can overcome the difficulty in the congenital diffusion that macromole such as general somatomedin can't overcome, and has chemical induction (chemo-attraction) function, can effectively borrow dynamic chmotropism abduction mechanism guiding endotheliocyte to permeate system so far, and the signal pipeline is different with general somatomedin in the born of the same parents that it caused, and this comprises that the biodegradability base material of phospholipid has the usefulness that can promote revascularization with the somatomedin matched combined.Superiority of the present invention is:
(1) interpolation of the interior active phospholipid of carrier can promote the effect of revascularization;
(2) active phospholipid makes an addition to the biodegradability base material with the liposome kenel, and to be prepared into the system that tool promotes revascularization usefulness, this mode is easy, and the usefulness of handling ease and system is stable;
(3) because phospholipid is the bipolarity molecule, can combine with hydrophilic and hydrophobic substrates simultaneously, such characteristic can increase the multiformity of application;
(4) because phospholipid is bipolarity molecule and critical liposome concentration (critical micelle concentration that they are special; CMC), when they are present in base material with the liposome kenel, can reach the effect that continues release, and can change its release profiles to meet various demands clinically by the processing and the modification of base material.
The present invention relates to a kind of Biodegradable composite material that promotes revascularization, it comprises: (a) biological degradable material, and (b) monophosphatide are distributed in the above-mentioned biological degradable material with the kenel of liposome.
The character of phospholipid used herein is stable than somatomedin, and individualism stably, and its molecular weight is little, approximately has only between 300～400, and the diffusion rate between tissue in the matter is very fast.In fact, these phospholipid by secretion of platelet, can attract endotheliocyte to move and stimulating growth when platelet activation, can also suppress the growth of smooth muscle cell and fibroblast to prevent the generation of tissue fibering simultaneously.In addition, phospholipid is that the main component of cell membrane also is important signal transmitter in the cell, almost signal pipeline in all higher mammal cells and signal transmitter are identical, because its universality and molecular weight are little, make it that problem of immune compatibility can not take place.
Be applicable to that phospholipid of the present invention comprises (but being not limited thereto): nerve sheath amine alcohol-1-phosphide (S-1-P; Sphingosine-1-Phosphate), haemolysis phosphate acid ester (L-P-A; Lysophosphatidic acid) or other by platelet or the isolated phosphide of cell membrane, wherein (S-1-P) is preferable with nerve sheath amine alcohol-1-phosphide again.Since nerve sheath amine alcohol-1-phosphide (S-1-P) be the bipolarity molecule with and higher critical liposome concentration, in composite, exist and can continue to discharge the S-1-P monomer and quickly diffuse in the tissue with the kenel of monolayer liposome.Therefore, this complex attracts endotheliocyte to move the growth of promotion local vascular and the healing of wound constantly.In addition, S-1-P receptor (S-1-P receptor) is G-protein-coupled receptor (G-protein-coupled receptor), and it comes activating cell and transmitting signals by the various dissimilar G-albumen of activation; Relative, the activated channel of general protein-based somatomedin then is to be main with receptor tyrosine kinase (RTK, ReceptorTyrosine Kinase).These two kinds of approach are independent respectively but may have some interactional mechanism, therefore composite of the present invention is except phospholipid, also can add suitable somatomedin according to different needs, differentiation by the phospholipid derivative stimulating endothelial cell, can produce the effect of taking advantage of, the effect of more effective enhancing revascularization of adding with single somatomedin effect.
Be applicable to that biological degradable material of the present invention comprises (but being not limited thereto): gelatin (Gelatin), collagen protein (Collagen), chitin (Chitin), remove acetyl chitin (Chitosan), aminoglucose polysaccharide (Glucosaminoglycans), chondroitin sulfate (Chondroitin sulfates), hyaluronic acid (Hyaluronic acid) or other protein-based class extracellular matrix (ECM proteins), alginate (Alginates), starch/modified starch (Starch/modified starch), alginic acid (Carragenam)/salt, pectin (Pectins), other polysaccharide class extracellular matrix (ECM polysaccharides), polyactide-Acetic acid, hydroxy-, bimol. cyclic ester acid (PLGA; Polylactide glycolide acid) and take from the material of bio-tissue.The preferably can use natural high molecular composite material, and it has the advantage in hypotoxicity, biodegradable, the medical application of biocompatibility higher organism.
In preferable embodiment of the present invention, system uses collagen protein and polysaccharide compound as biological degradable material, has the d cell receptor, special cells affinity aglucon (cellaffinity ligand), dual different enzyme decomposes approach and decomposition rate, so can prolong the resolving time, and can attract some cell to attach.Moreover the system that collagen protein/polysaccharide compound composite constitutes and the compatibility of S-1-P liposome are very high, and S-1-P mode very uniformly is distributed in the material.In addition, collagen protein/polysaccharide compound composite has multiple hydrophilic (hydrophilic) functional group, can catch a large amount of hydrones, when becoming glue, can not cause the dehydrating condensation phenomenon, can prevent that S-1-P and somatomedin from being discharged fast because of dehydrating condensation.
Manufacture method of the present invention, system is dissolving phospholipid and be coated on chamber wall with the coating method of very big area earlier, utilize the temperature difference and ultrasound concussion to be prepared into the liposome of phospholipid in the mode that is separated then, mix the liposome and the Biodegradable composite material of this phospholipid at last, co-production becomes to promote the Biodegradable composite material or the base material element of revascularization.Detailed step comprises: (a) with dissolution with solvents phospholipid, be placed in the container; (b) remove solvent in the distilling under reduced pressure mode, make phospholipid be attached to the wall of container; (c) in container, add phosphoric acid buffer saline solution (PBS; Phosphate BufferedSaline), and with its IQF; (d) with ultrasound concussion container, make phospholipid in the phosphoric acid buffer saline solution, form liposome; And (e) liposome of phospholipid is mixed with biological degradable material, to form the Biodegradable composite material that promotes revascularization.
The employed solvent of step (a) is a polar organic solvent, is generally alcohols, for example methanol or ethanol.The preferable liquid nitrogen that uses of step (c) cools off it.In step (e), can earlier biological degradable material be processed into the base material of various kenels such as the block base material of glue, porous, film substrate, fiber base material, tubulose base material, paste base material after, use with mixed interpolation of phospholipid again.Perhaps, also can earlier phospholipid be mixed and make an addition in the decomposability material, be reprocessed into above-mentioned various kenel, for example can be in advance with liposome and the collagen solution (0.6-1.0w/v of S-1-P, in 1% organic acid solvents such as 1% acetic acid or lactic acid) mix after, make the porous composite in cryodesiccated mode again.The composite of glue kenel can be implanted in the animal body with hypodermic mode, and other kenel such as porous substrate can adopt modus operandi that it is implanted in skin corium and subcutaneous tissue.
In addition, composite of the present invention also can add other protein-based somatomedin, inhibitive factor or stimulating factor (cytokines) and merge use.Somatomedin commonly used comprises (but being not limited thereto): the fibrous tissue growth factor (FGF; Fibroblast growth factor), placental growth factor (Placental growth factor), degeneration somatomedin (Transforming growthfactor), angiogenine (Angiogenin), interleukin 8 (Interleukin-8), hepatocyte growth factor (Hepatocyte growth factor), granulocyte colony stimulating factor (Granulocyte colony-stimulating factor), platelet source endothelial cell growth factor (ECGF) (Platelet-derived endothelial cell growth factor) etc.Inhibitive factor commonly used comprises (but being not limited thereto): interferon-ALPHA (Interferon α), change shape somatomedin (Transforming growth factor β), thrombospondin-1 (Thrombospondin-1), angiostatin (Angiostatin), the placenta accreta platelet factor 4 (Placental proliferin-related, Platelet factor 4) of being correlated with, platelet factor 4 (Platelet factor 4), genistein (Genistein), metalloprotein enzyme inhibition factor (Metallo-proteinase inhibitor), prolactin antagonist 16-kd fragment (Prolactin 16-kd fragment) etc.
The composite of the promotion revascularization that the present invention in sum, developed has biomedical material and uses upward required characteristic and advantage:
1. this composite has hypotoxicity, high biodegradable, biocompatibility etc., has the advantage on the biomedical applications;
2. the glue system of this composite formation has class extracellular matrix (ExtracellularMatrix-like) structure, is fit to cell and attaches and growth;
3. this composite has the function that promotes revascularization;
4. this composite is preserved conveniently, and preparation is simple, can be reduced in the course of processing for active destruction of carrier organism and influence degree;
5. described phospholipid is the bipolarity molecule, can combine with hydrophilic and hydrophobic substrates simultaneously, and this specific character can increase the multiformity of application;
6. can add bioactie agent (comprising somatomedin and medicine) more in addition, and in the application such as the drug release under being applied to, organizational project.
Description of drawings
For above and other objects of the present invention, feature and advantage can be become apparent, cited below particularlyly go out preferred embodiment, and conjunction with figs., be described in detail below:
Fig. 1 is the result after the mouse organization's section among the embodiment 1 adds HE dyeing, and wherein (a) is that bFGF and S-1-P merge use, (b) use separately for bFGF, and (c) be the control experiment group of PBS;
Fig. 2 is the mean vascular counting on the tissue slice, and blood vessel quantity is bFGF/S-1-P＞bFGF＞PBS, wherein PBS in regular turn
*Do not add glue.
The specific embodiment
Embodiment 1: glue system, collagen-free albumen
The composite of present embodiment comprises the natural polymer complex of biodegradabilities such as poly-polysaccharide body (chitin) and phospholipid S-1-P, and its method for making is as follows:
At first, S-1-P with dissolve with methanol and pour in the round-bottomed flask, is coated on S-1-P the flask inner surface and is evacuated to bone dry with vacuum distillation apparatus then.Then, add an amount of PBS, and with the liquid nitrogen IQF.Handle with the ultrasound concussion in room temperature, make the S-1-P dissolving (formation liposome) on the flask walls.At last, the liposome of S-1-P is mixed with the chitosan glue, and somatomedin bFGF is added in the composite.The composite of gained is implanted in the mouse body in hypodermic mode.
Fig. 1 adds result after the HE dyeing for tissue slice.Mouse was put to death behind the subcutaneous injection glue on the 15th day, takes out after the implant through fixing, and embedding, section, the size and the kenel of blood vessel observed in the dyeing back under 200 times the visual field.(a) be that bFGF and S-1-P merge use, can find that the radius of blood vessel and quantity all obviously increase than independent use of bFGF (b) and control experiment group (c), the arrow place is the blood vessel position.Fig. 2 is the mean vascular counting on the tissue slice, and blood vessel quantity is bFGF/S-1-P＞bFGF＞PBS in regular turn, and is consistent with the observed result of Fig. 1.
Embodiment 2: there is collagen protein in the glue system
The composite of present embodiment comprises protein (collagen protein), the natural polymer complex of biodegradabilities such as poly-polysaccharide body (chitin) and phospholipid S-1-P.
At first, with S-1-P with dissolve with methanol and pour in the round-bottomed flask.Then, with vacuum distillation apparatus S-1-P is coated on the flask inner surface and is evacuated to bone dry.Then, add an amount of PBS, and with the liquid nitrogen IQF.Handle with the ultrasound concussion in room temperature, make the S-1-P dissolving (formation liposome) on the tube wall.At last, the thermo-responsive compound glue of liposome and the chitosan-collagen protein of S-1-P is mixed, obtain containing the composite of collagen protein, chitin and phospholipid S-1-P.
Embodiment 3: porous substrate
The composite of present embodiment comprises the cellular collagen protein, the natural polymer complex of biodegradabilities such as phospholipid S-1-P.
At first, S-1-P with dissolve with methanol and pour in the round-bottomed flask, is coated on S-1-P the flask inner surface and is evacuated to bone dry with vacuum distillation apparatus then.Then, add an amount of PBS, and with the liquid nitrogen IQF.Handle with the ultrasound concussion in room temperature, make the S-1-P dissolving (formation liposome) on the tube wall.At last, the composite mixing that the liposome of S-1-P is added cellular collagen protein (collagenporous sponge).
Though the present invention discloses as above with preferred embodiment, so it is not in order to qualification the present invention, any person skilled in the art, and without departing from the spirit and scope of the present invention, change of having done and retouching all should be within the scope of the present invention.
Claims (21)
1. Biodegradable composite material that promotes revascularization, it comprises:
One biological degradable material; And
Monophosphatide is distributed in the above-mentioned biological degradable material with the kenel of liposome.
2. the Biodegradable composite material of promotion revascularization as claimed in claim 1, wherein said biological degradable material is selected from: gelatin, collagen protein, polysaccharide compound, chitin, remove acetyl chitin, aminoglucose polysaccharide, chondroitin sulfate, hyaluronic acid, alginate, starch/modified starch, alginic acid/salt, pectin, polyactide-Acetic acid, hydroxy-, bimol. cyclic ester acid and mixture thereof.
3. the Biodegradable composite material of promotion revascularization as claimed in claim 1, wherein said biological degradable material are collagen protein and polysaccharide compound.
4. the Biodegradable composite material of promotion revascularization as claimed in claim 1, wherein said phospholipid are nerve sheath amine alcohol-1-phosphide.
5. the Biodegradable composite material of promotion revascularization as claimed in claim 1, wherein said phospholipid are the haemolysis phosphate acid ester.
6. the Biodegradable composite material of promotion revascularization as claimed in claim 1, wherein said phospholipid are by platelet or the isolated phosphide of cell membrane.
7. the Biodegradable composite material of promotion revascularization as claimed in claim 1, wherein said Biodegradable composite material have following wherein a kind of kenel: glue, the block base material of porous, film substrate, fiber base material, tubulose base material or paste base material.
8. the Biodegradable composite material of promotion revascularization as claimed in claim 1, it further comprises somatomedin, inhibitive factor or stimulating factor.
9. the Biodegradable composite material of promotion revascularization as claimed in claim 8, wherein said somatomedin is selected from: the fibrous tissue growth factor, placental growth factor, degeneration somatomedin, angiogenine, interleukin 8, hepatocyte growth factor, granulocyte colony stimulating factor and platelet source endothelial cell growth factor (ECGF).
10. the Biodegradable composite material of promotion revascularization as claimed in claim 8, wherein said inhibitive factor is selected from: interferon-ALPHA, commentaries on classics shape somatomedin, thrombospondin-1, angiostatin, the relevant platelet factor 4 of placenta accreta, platelet factor 4, genistein, metalloprotein enzyme inhibition factor and prolactin antagonist 16-kd fragment.
11. the manufacture method of the Biodegradable composite material of the described promotion revascularization of claim 1, it comprises the following steps:
(a), be placed in the container with dissolution with solvents phospholipid;
(b) remove this solvent in the distilling under reduced pressure mode, make this phospholipid be attached to the wall of this container;
(c) in this container, add the phosphoric acid buffer saline solution, and with its IQF;
(d) shake this container with ultrasound, make this phospholipid in this phosphoric acid buffer saline solution, form liposome; And
(e) liposome with this phospholipid mixes with biological degradable material, to form the Biodegradable composite material that promotes revascularization.
12. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 11, wherein said solvent are polar organic solvent.
13. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 12, wherein said solvent are alcohols.
14. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 11, wherein said phospholipid are nerve sheath amine alcohol-1-phosphide, haemolysis phosphate acid ester or by platelet or the isolated phosphide of cell membrane.
15. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 11, wherein the IQF described in the step (c) is to use liquid nitrogen.
16. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 11, wherein said biological degradable material is selected from: gelatin, collagen protein, polysaccharide compound, chitin, remove acetyl chitin, aminoglucose polysaccharide, chondroitin sulfate, hyaluronic acid, alginate, starch/modified starch, alginic acid/salt, pectin, polyactide-Acetic acid, hydroxy-, bimol. cyclic ester acid and mixture thereof.
17. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 16, wherein said biological degradable material are collagen protein, polysaccharide compound.
18. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 11, wherein said Biodegradable composite material have following wherein a kind of kenel: glue, the block base material of porous, film substrate, fiber base material, tubulose base material or paste base material.
19. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 11, wherein step (e) further comprises: add somatomedin, inhibitive factor or stimulating factor in this biological degradable material.
20. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 19, wherein said somatomedin is selected from: the fibrous tissue growth factor, placental growth factor, degeneration somatomedin, angiogenine, interleukin 8, hepatocyte growth factor, granulocyte colony stimulating factor and platelet source endothelial cell growth factor (ECGF).
21. the manufacture method of the Biodegradable composite material of promotion revascularization as claimed in claim 19, wherein said inhibitive factor is selected from: interferon-ALPHA, commentaries on classics shape somatomedin, thrombospondin-1, angiostatin, the relevant platelet factor 4 of placenta accreta, platelet factor 4, genistein, metalloprotein enzyme inhibition factor and prolactin antagonist 16-kd fragment.
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| CN 01145295 CN1429628A (en) | 2001-12-31 | 2001-12-31 | Biodegradable composite material for promoting regeneration of blood vessel and its manufacturing method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109395095A (en) * | 2017-09-18 | 2019-03-01 | 武汉原生原代生物医药科技有限公司 | In vivo with biomembrane and its preparation method and application |
| CN112843072A (en) * | 2021-01-12 | 2021-05-28 | 山东省科学院生物研究所 | Use of phospholipids for regulating angiogenesis |
| CN113827768A (en) * | 2021-09-22 | 2021-12-24 | 苏州大学附属第二医院 | Preparation method of bionic scaffold carrying lysophosphatidic acid nanoparticles |
| CN113842504A (en) * | 2021-09-22 | 2021-12-28 | 苏州大学附属第二医院 | Preparation method of multifunctional electrospinning bracket for bone regeneration |
-
2001
- 2001-12-31 CN CN 01145295 patent/CN1429628A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109395095A (en) * | 2017-09-18 | 2019-03-01 | 武汉原生原代生物医药科技有限公司 | In vivo with biomembrane and its preparation method and application |
| CN109395095B (en) * | 2017-09-18 | 2021-09-24 | 武汉原生原代生物医药科技有限公司 | In vivo biological membrane and preparation method and application thereof |
| CN112843072A (en) * | 2021-01-12 | 2021-05-28 | 山东省科学院生物研究所 | Use of phospholipids for regulating angiogenesis |
| CN113827768A (en) * | 2021-09-22 | 2021-12-24 | 苏州大学附属第二医院 | Preparation method of bionic scaffold carrying lysophosphatidic acid nanoparticles |
| CN113842504A (en) * | 2021-09-22 | 2021-12-28 | 苏州大学附属第二医院 | Preparation method of multifunctional electrospinning bracket for bone regeneration |
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