CN117899271A - Composite tissue engineering scaffold with multi-stage pore structure and preparation method thereof - Google Patents
Composite tissue engineering scaffold with multi-stage pore structure and preparation method thereof Download PDFInfo
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- CN117899271A CN117899271A CN202410085291.2A CN202410085291A CN117899271A CN 117899271 A CN117899271 A CN 117899271A CN 202410085291 A CN202410085291 A CN 202410085291A CN 117899271 A CN117899271 A CN 117899271A
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- 238000002360 preparation method Methods 0.000 title claims abstract description 11
- DLYUQMMRRRQYAE-UHFFFAOYSA-N tetraphosphorus decaoxide Chemical compound O1P(O2)(=O)OP3(=O)OP1(=O)OP2(=O)O3 DLYUQMMRRRQYAE-UHFFFAOYSA-N 0.000 claims abstract description 40
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- 230000007547 defect Effects 0.000 claims description 10
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Abstract
The invention belongs to the technical field of bone tissue engineering, and in particular relates to a preparation method of a composite tissue engineering scaffold with a multi-stage pore structure, which comprises the following steps: (1) Uniformly stirring each raw material SiO 2,CaO,Na2O,P2O5 to prepare mixed sol, and preparing xerogel powder with a submicron structure through a sol-gel conversion process; (2) Fully mixing xerogel powder with a fluid pore-forming medium, pouring the mixture into a mold, curing the mixed system by controlling the temperature, and demolding; (3) Removing the pore-forming medium, and sintering to obtain the three-dimensional porous bioactive glass bracket with a submicron/micron two-stage pore structure; (4) In the pores of the three-dimensional porous bioactive glass scaffold, a nanofiber network structure is synthesized in situ by utilizing a bacterial fermentation technology, so that the composite tissue engineering scaffold which is close to the highly bionic human natural bone and has a multi-stage pore structure is obtained.
Description
Technical Field
The invention belongs to the technical field of bone tissue engineering, and particularly relates to a composite tissue engineering scaffold with a multi-stage pore structure and a preparation method thereof.
Background
Bone is the largest tissue organ of the human body and bears the important responsibility of life activities, but the bone is most easy to cause defects, and millions of bone tissue defect patients need to receive surgical treatment every year. The incidence rate of the orthopaedics diseases is high (15-20% of population appears with different degrees of orthopaedics diseases each year), the disability rate of orthopaedics wounds is high (the most common cause of disability of people under 45 years old), the medical cost is high (the U.S. is up to 500 hundred million dollars each year), and huge household, social and economic burdens are caused. Bone grafting has become the most demanding implant next to blood transfusion. How to break the 'password' of the human bone, how to find more better bone tissue regeneration and repair materials, and the method is a re-creation and health for repairing life of human beings, and has become the power for common pursuit and cumin tiredness research of a plurality of scientists worldwide. The research and development of new technology, new theory and new method of the orthopedic repair material have urgent scientific significance, social value and economic benefit.
Currently, the application of tissue engineering materials in hard tissues is increasingly emphasized, namely bones, cartilage, oral cavity and the like. Bones, in particular joints, are the tissue organs of the human body which are mainly subjected to load, which means that the repair material of the defect should have high strength. Materials used for hard tissue repair and replacement are mainly metals and alloys, bioceramics, polymers and composites. With the development of tissue engineering, in the aspect of bone tissue engineering, cells can be planted on a degradable biomaterial three-dimensional scaffold, cultured in vivo or in vitro, and then implanted into a defect site to repair and regenerate bone tissue. The cell growth factors such as bone morphogenetic protein are compounded on the bracket to induce the regeneration of new bone with natural bone function, which is a struggle target of new generation bone tissue engineering. Bone is a natural compound composed of hydroxyapatite and collagen, 10-30% of which is composed of a hard outer layer, i.e. dense bone; internally, 30-90% is cancellous bone. Because of the differences in proportions of dense bone and cancellous bone, and the complexity of the geometry of the constituent bone tissue, the mechanical properties of bone are quite different, and it is therefore not easy to design and manufacture an "ideal bone scaffolding material". Therefore, how to obtain a highly bionic artificial bone scaffold material close to the structures of various scales of human natural bones, and enabling the in vivo dynamic degradation and degeneration of the scaffold to be matched with the growth of new tissues in terms of physical scale and mechanical properties is a key problem faced by the bone regeneration repair technology.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide a preparation method of a composite tissue engineering scaffold with a multi-stage pore structure, which adopts micron and submicron secondary sol-gel transformation to construct porous bioactive glass, and synthesizes nano-cellulose in situ on the basis of the structure to form the composite scaffold of micro-nano multi-stage pore structure bioactive glass-bacterial cellulose (BAG-BC).
In order to achieve the above purpose, the invention adopts the following technical scheme:
the preparation method of the composite tissue engineering scaffold with the multi-stage pore structure comprises the following steps:
(1) Uniformly stirring each raw material SiO 2,CaO,Na2O,P2O5 to prepare mixed sol, and preparing xerogel powder with a submicron structure through a sol-gel conversion process;
(2) Fully mixing xerogel powder with a fluid pore-forming medium, pouring the mixture into a mold, curing the mixed system by controlling the temperature, and demolding;
(3) Removing the pore-forming medium, and sintering at 650-680 ℃ to obtain a three-dimensional porous bioactive glass bracket with a submicron/micron two-stage pore structure;
(4) In the pores of the three-dimensional porous bioactive glass scaffold, the nano fiber network structure is synthesized in situ by utilizing a bacterial fermentation technology, and the composite tissue engineering scaffold with the multi-stage pore structure is obtained.
In a preferred example, the mole percentage of each raw material SiO 2,CaO,Na2O,P2O5 is 45 to 50 mole percent, siO 2,24～26mol％CaO,21～24mol％Na2O,1.5～2mol％P2O5.
In a preferred example, the mole percent between the starting materials SiO 2,CaO,Na2O,P2O5 is 49.15 mole percent, siO 2,25.80mol％CaO,23.33mol％Na2O,1.72mol％P2O5 in turn.
In a preferred example, the pore-forming medium is paraffin.
In a preferred example, the shape of the mold is customized according to the physical shape of the bone defect site.
In a preferred example, the step of removing the pore-forming medium comprises drying and firing sequentially.
Based on one general inventive concept, another object of the present invention is to protect the composite tissue engineering scaffold with a multi-stage pore structure manufactured by the above manufacturing method.
Firstly, preparing xerogel particles with target components by a sol-gel technology, wherein the xerogel particles have a submicron structure; then fully mixing xerogel particles with a pore-forming medium according to the low melting point or property transformation characteristic of the specific pore-forming medium, solidifying the mixed system by controlling the temperature, demoulding, and sintering to obtain the bioactive glass porous material with a submicron/micron two-stage pore structure; on the basis of the two-stage pore structure bioactive glass porous structure, the nano-structure fiber network structure is synthesized in situ in the pores of the porous structure bioactive glass by utilizing a bacterial fermentation technology, so that the composite tissue engineering scaffold with the multi-stage pore structure is obtained.
Compared with the prior art, the invention has the following advantages and positive effects:
Structurally, the pore diameter, pore wall thickness, specific surface area and the like of the macropores and the pinholes of the bioactive glass and the communication holes are controlled through research to simulate a lamellar porous structure of bone tissue staggering, and a nanofiber network structure of a bone extracellular matrix is simulated by a natural nano structure of nanocellulose;
the components simulate the element composition and the material distribution of bone tissues by using calcium and phosphorus elements and the like of the bioactive glass, and the structures of the bioactive glass such as organic-inorganic phase distribution and the like.
The highly bionic artificial bone scaffold material which is close to the structures of various scales of the natural bone of the human is obtained, and the in vivo dynamic degradation and degeneration of the scaffold and the growth of the new tissue can be matched in physical scale and mechanical property so as to regenerate and repair the bone.
Drawings
FIG. 1 is an in situ synthesis process of bacterial cellulose in a bioactive glass three-dimensional porous scaffold.
Detailed Description
The invention will be further described with reference to specific embodiments for the purpose of making the objects, technical solutions and advantages of the invention more apparent, but the invention is not limited to these examples. It should be noted that, on the premise of no conflict, new embodiments may be formed by any combination of the embodiments or technical features described below. In the present invention, the equipment, materials, etc. used are commercially available or commonly used in the art, unless otherwise specified. The methods in the following examples are conventional in the art unless otherwise specified.
The terms "comprising," "including," "containing," or any other variation thereof, as used herein, are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, step, method, article, or apparatus.
When an equivalent, concentration, or other value or parameter is expressed as a range, preferred range, or a range bounded by a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any upper range limit or preferred value and any lower range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when ranges of "1 to 5" are disclosed, the described ranges should be construed to include ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. When a numerical range is described herein, unless otherwise indicated, that range is intended to include its endpoints and all integers and fractions within that range.
Specific embodiments of the present invention are described in detail below.
Example 1
The preparation method of the composite tissue engineering scaffold with the multi-stage pore structure comprises the following steps:
(1) Uniformly stirring each raw material SiO 2,CaO,Na2O,P2O5 to prepare mixed sol, and preparing xerogel powder with a submicron structure through a sol-gel conversion process;
wherein, the mol percent of each raw material SiO 2,CaO,Na2O,P2O5 is 49.15mol percent, siO 2,25.80mol％CaO,23.33mol％Na2O,1.72mol％P2O5;
(2) Fully mixing xerogel powder with a fluid pore-forming medium (paraffin), pouring into a mould customized according to the physical shape of a bone defect part, solidifying the mixed system by controlling the temperature, and demoulding;
(3) Removing a pore-forming medium through drying, roasting and other processes, and then sintering at 670 ℃ to obtain the three-dimensional porous bioactive glass bracket with a submicron/micron two-stage pore structure;
(4) In the pores of the three-dimensional porous bioactive glass scaffold, the nano fiber network structure is synthesized in situ by utilizing a bacterial fermentation technology, and the composite tissue engineering scaffold with the multi-stage pore structure is obtained.
Example 2
The preparation method of the composite tissue engineering scaffold with the multi-stage pore structure comprises the following steps:
(1) Uniformly stirring each raw material SiO 2,CaO,Na2O,P2O5 to prepare mixed sol, and preparing xerogel powder with a submicron structure through a sol-gel conversion process;
wherein, the mol percent of each raw material SiO 2,CaO,Na2O,P2O5 is 45.21mol percent, siO 2,24.72mol％CaO,21.43mol％Na2O,1.56mol％P2O5;
(2) Fully mixing xerogel powder with a fluid pore-forming medium (paraffin), pouring into a mould customized according to the physical shape of a bone defect part, solidifying the mixed system by controlling the temperature, and demoulding;
(3) Removing a pore-forming medium through drying, roasting and other processes, and sintering at 650 ℃ to obtain the three-dimensional porous bioactive glass bracket with a submicron/micron two-stage pore structure;
(4) In the pores of the three-dimensional porous bioactive glass scaffold, the nano fiber network structure is synthesized in situ by utilizing a bacterial fermentation technology, and the composite tissue engineering scaffold with the multi-stage pore structure is obtained.
Example 3
The preparation method of the composite tissue engineering scaffold with the multi-stage pore structure comprises the following steps:
(1) Uniformly stirring each raw material SiO 2,CaO,Na2O,P2O5 to prepare mixed sol, and preparing xerogel powder with a submicron structure through a sol-gel conversion process;
Wherein the mol percent of each raw material SiO 2,CaO,Na2O,P2O5 is 47.52mol percent, siO 2,24.21mol％CaO,22.85mol％Na2O,1.89mol％P2O5;
(2) Fully mixing xerogel powder with a fluid pore-forming medium (paraffin), pouring into a mould customized according to the physical shape of a bone defect part, solidifying the mixed system by controlling the temperature, and demoulding;
(3) Removing a pore-forming medium through drying, roasting and other processes, and sintering at 680 ℃ to obtain the three-dimensional porous bioactive glass bracket with a submicron/micron two-stage pore structure;
(4) In the pores of the three-dimensional porous bioactive glass scaffold, the nano fiber network structure is synthesized in situ by utilizing a bacterial fermentation technology, and the composite tissue engineering scaffold with the multi-stage pore structure is obtained.
The above embodiments are merely preferred embodiments of the present invention, and any simple modification, modification and substitution changes made to the above embodiments according to the technical substance of the present invention are all within the scope of the technical solution of the present invention.
Claims (7)
1. The preparation method of the composite tissue engineering scaffold with the multi-stage pore structure is characterized by comprising the following steps:
(1) Uniformly stirring each raw material SiO 2,CaO,Na2O,P2O5 to prepare mixed sol, and preparing xerogel powder with a submicron structure through a sol-gel conversion process;
(2) Fully mixing xerogel powder with a fluid pore-forming medium, pouring the mixture into a mold, curing the mixed system by controlling the temperature, and demolding;
(3) Removing the pore-forming medium, and sintering at 650-680 ℃ to obtain a three-dimensional porous bioactive glass bracket with a submicron/micron two-stage pore structure;
(4) In the pores of the three-dimensional porous bioactive glass scaffold, the nano fiber network structure is synthesized in situ by utilizing a bacterial fermentation technology, and the composite tissue engineering scaffold with the multi-stage pore structure is obtained.
2. The method for preparing the composite tissue engineering scaffold with the multi-stage pore structure according to claim 1, wherein the mole percentage of each raw material SiO 2,CaO,Na2O,P2O5 is 45-50 mol% and SiO 2,24～26mol％CaO,21～24mol％Na2O,1.5～2mol％P2O5.
3. The method for preparing the composite tissue engineering scaffold with the multi-stage pore structure according to claim 1, wherein the mole percentage of each raw material SiO 2,CaO,Na2O,P2O5 is 49.15mol% and the mole percentage of each raw material SiO 2,25.80mol％CaO,23.33mol％Na2O,1.72mol％P2O5 is 49.15 mol%.
4. The method for preparing a composite tissue engineering scaffold with a multi-stage pore structure according to claim 1, wherein the pore-forming medium is paraffin.
5. The method for preparing a composite tissue engineering scaffold with a multi-stage pore structure according to claim 1, wherein the shape of the mold is customized according to the physical shape of the bone defect site.
6. The method for preparing a composite tissue engineering scaffold with a multi-stage pore structure according to claim 1, wherein the step of removing the pore-forming medium comprises drying and roasting in sequence.
7. The composite tissue engineering scaffold with a multi-stage pore structure prepared by the preparation method of any one of claims 1 to 6.
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