CN115737926A - Living traditional Chinese medicine bracket for wound repair and preparation method thereof - Google Patents
Living traditional Chinese medicine bracket for wound repair and preparation method thereof Download PDFInfo
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Abstract
The invention provides a living traditional Chinese medicine bracket for wound repair and a preparation method thereof, wherein the living traditional Chinese medicine bracket comprises the following components: taking the aqueous phase solution of the cross-linking agent as an internal phase and the mixed dispersion liquid containing the living microalgae and the traditional Chinese medicine molecules as an external phase, extruding hollow fibers in the air, and obtaining living traditional Chinese medicine hydrogel hollow fiber elements after cross-linking and curing; printing the hollow fiber element on a flat plate by a 3D printing mode to obtain a living traditional Chinese medicine hydrogel fiber scaffold with a three-dimensional communicated porous structure, and then performing crosslinking and solidification to obtain the three-dimensional living traditional Chinese medicine scaffold. The traditional Chinese medicine molecules and the living microalgae are introduced to endow the stent with dual effects of continuous oxygen supply and drug release, the traditional Chinese medicine molecules can promote the activity of microalgae cells in the living stent and promote the oxygen production activity of the microalgae cells to be improved, and the microalgae cells can be gradually released along with the degradation of the stent, so that the wound healing speed and quality are remarkably improved, and the traditional Chinese medicine molecules are widely applicable to the repair of various chronic tissue defects.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to a living traditional Chinese medicine bracket for wound repair and a preparation method thereof.
Background
Wound healing is a major concern in the medical and health field worldwide, as it frequently occurs in daily life and surgery. In general, wound healing is a highly coordinated dynamic process that mainly comprises three phases: inflammatory, proliferative and tissue maturation stages. Chronic wounds often occur in large areas of tissue defects or in situations where the patient is in a diseased condition (e.g., diabetes). Accordingly, a number of wound dressings and biological scaffold materials for wound healing are being developed. Among them, 3D printing scaffolds are attracting much attention due to their unique advantages, such as freely customizable geometries, porous structures, and various bioactive components. These porous scaffolds not only provide structural support for cell migration, facilitate the growth of new tissue, but also serve as effective carriers for the delivery of various bioactive molecules to the wound. However, currently developed wound dressing materials tend to be inefficient to load, requiring frequent dressing changes until the wound is fully healed. While some stimulus responsive delivery systems can achieve drug release on demand over a short period of time (typically only a few hours or days), it is difficult to maintain the required drug concentration over the course of a long-term healing.
For thousands of years, traditional Chinese medicine has proved to have great practical value in preventing and treating various diseases. Wherein, the notoginseng is one of the most commonly used Chinese medicinal materials, and the panax notoginseng saponins are main active substances extracted from the root of the notoginseng plant, and have the functions of inhibiting inflammation, promoting angiogenesis and promoting the healing of diabetic wounds. In addition, the microalgae is a novel natural biological material, and has unique nutritive value and high-efficiency photosynthetic oxygen production.
In summary, a living traditional Chinese medicine bracket prepared by combining the materials and adopting a 3D printing mode and used for wound repair is urgently needed.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides a living traditional Chinese medicine bracket for wound repair and a preparation method thereof. According to the invention, by combining a micro-fluidic spinning technology and a 3D printing technology, traditional Chinese medicine molecules of panax notoginseng saponins and living microalgae are introduced into a tissue engineering scaffold system, and panax notoginseng drug molecules can promote the activity of microalgae cells in the living scaffold, promote the oxygen production activity of the microalgae to be improved, and gradually release the microalgae along with the degradation of the scaffold, so that the wound oxygen deficiency is relieved, the angiogenesis and skin regeneration are accelerated, the speed and quality of wound healing are effectively improved, and the tissue engineering scaffold is widely suitable for repairing various chronic tissue defects.
In order to realize the purpose, the invention adopts the following technical scheme:
a living traditional Chinese medicine bracket for wound repair and a preparation method thereof comprise the following steps:
(1) Constructing a one-dimensional living traditional Chinese medicine hydrogel hollow fiber element:
designing and constructing a nested incident channel microfluidic device, taking an aqueous phase solution of a cross-linking agent as an internal phase and a mixed dispersion liquid containing living microalgae and traditional Chinese medicine molecules as an external phase, extruding a preliminarily formed hollow fiber in the air by using the microfluidic device, and irradiating, crosslinking and curing by ultraviolet laser to obtain a living traditional Chinese medicine hydrogel hollow fiber element with uniform size, continuity and stability;
(2) Preparing a three-dimensional living traditional Chinese medicine bracket by microfluidic bioprinting:
directly printing the hollow fiber elements prepared in the step (1) on a cleaning flat plate in a 3D printing mode to form a three-dimensional living traditional Chinese medicine hydrogel fiber support which is stacked layer by layer and arranged in a cross mode and has a three-dimensional communicated porous structure, and then irradiating, crosslinking and curing by ultraviolet laser to obtain the three-dimensional living traditional Chinese medicine support;
the size of the channel of the hollow fiber is regulated and controlled by adjusting the diameter of the microfluidic channel and the flow rates of the internal phase fluid and the external phase fluid;
regulating and controlling the bioactivity of the hollow fiber by changing the composition ratio of the living microalgae and the traditional Chinese medicine molecules in the mixed dispersion liquid;
the porous structure, the macroscopic appearance and the size of the three-dimensional living traditional Chinese medicine bracket are regulated and controlled by adjusting the 3D printing program.
In order to optimize the technical scheme, the specific measures adopted further comprise:
further, in the step (1), the nested incident channel microfluidic device is assembled by a glass capillary, a glass slide, a sample application needle and quick-drying glue; wherein, the glass capillary is assembled by coaxially nesting an external phase capillary and an internal phase capillary.
Furthermore, the diameter of the external phase capillary tube is 250-1000 μm, and the diameter of the internal phase capillary tube is 100-200 μm; the inner diameter and the outer diameter of the hollow fiber are adjusted by changing the diameters of the inner capillary tube and the outer capillary tube.
Further, in the step (1), the inner phase is a mixed solution of calcium chloride and polyethylene glycol, and the outer phase is a mixed dispersion of living microalgae, traditional Chinese medicine molecules, sodium alginate and methacrylic acid anhydrified gelatin (GelMA); the living microalgae is Chlorella, and the Chinese medicinal molecules are Panax notoginsenosides.
Further, in the mixed dispersion liquid, the concentration of chlorella is 0-1000 ten thousand/mL, the concentration of panax notoginseng saponins is 0-2000 mu g/mL, the concentration of sodium alginate is 0.5-2.5 w/v%, and the concentration of methacrylic anhydrization gelatin is 1-20 w/v%; the oxygen release performance and the tissue regeneration activity of the hollow fiber element are adjusted by respectively changing the amount of chlorella and panax notoginseng saponins within the concentration ranges of 0-1000 ten thousand/mL and 0-2000 mug/mL, and the formability and the mechanical strength of the hydrogel hollow fiber element are adjusted by respectively changing the amount of sodium alginate and methacrylic anhydrized gelatin within the concentration ranges of 0.5-2.5 w/v% and 1-20 w/v%.
Further, in the mixed solution, the concentration of calcium chloride is 0-4 w/v%, and the concentration of polyethylene glycol is 1-50 w/v%.
Further, in the step (1), the inner diameter of the hollow fiber element is 0-200 μm, the outer diameter is 200-1500 μm, and the aperture size of the three-dimensional living traditional Chinese medicine scaffold is 0-5000 μm.
Further, in the step (2), the hollow fiber elements are stacked in a manner of orthogonal, oblique or parallel stacking; the three-dimensional living traditional Chinese medicine support is in a cylinder, a cube or a composite polyhedron shape so as to match actual wound surfaces with different shapes and depths.
Further, in the steps (1) and (2), the crosslinking curing mode is ultraviolet irradiation crosslinking curing.
The traditional Chinese medicine molecules of the living body traditional Chinese medicine bracket obtained by the preparation method can obviously improve the activity of microalgae, improve the integral photosynthesis efficiency of the bracket, promote the adhesion, proliferation and migration of skin cells and relieve the anoxic state of the cells, and the bracket continuously releases oxygen and the traditional Chinese medicine molecules in the repair process of the wound surface in vivo, thereby effectively improving the healing speed and quality of the chronic wound surface.
The invention has the beneficial effects that:
1) The invention depends on the microfluidic technology, adopts the nested microfluidic chip to prepare the hollow fiber element of the living traditional Chinese medicine hydrogel, has simple structure and few building steps, does not need complex machining process, and has simple process and convenient operation; the size of the channel of the hollow fiber is regulated and controlled by adjusting the pipe diameter of a capillary in the micro-fluidic chip and the flow velocity of internal and external fluids; living microalgae proliferate in hydrogel containing Notoginseng radix medicinal molecules for a long time, generate more oxygen through photosynthesis, change its content can effectively regulate oxygen production rate and release amount, and relieve in vivo and in vitro cell anoxia state; in addition, the hydrogel matrix of the living body scaffold can be gradually degraded in a body fluid environment, released microalgae cells and pseudo-ginseng drug molecules are remained at a wound part, the effects of relieving an anoxic environment, promoting angiogenesis and accelerating wound healing are achieved, and the in-vivo and in-vitro tissue regeneration activity of the living body traditional Chinese medicine scaffold can be obviously improved by properly adjusting the content of the hydrogel matrix within a certain range.
2) According to the invention, the three-dimensional living traditional Chinese medicine bracket is prepared in a 3D printing mode, the size, the macroscopic appearance and the size, the shape and the connectivity of the pore channel of the bracket can realize accurate design and control, the efficient, rapid and repeatable preparation is realized, the artificial error is avoided, and the individualized requirements of different wound surfaces in practical application are met; the three-dimensional pore structure of the scaffold can provide an environment which is favorable for oxygen and nutrient substance transportation, thereby being more favorable for survival and propagation of microalgae cells in the scaffold. When the living body scaffold is implanted into a tissue defect part, the three-dimensional framework structure of the living body scaffold can provide temporary structural support for migration, adhesion, proliferation and differentiation of cells and guide the growth and reconstruction of new tissues.
3) According to the invention, the micro-fluidic spinning technology and the 3D printing technology are combined to introduce living microalgae and traditional Chinese medicine molecules into a hydrogel support system at the same time, so that the support has the dual characteristics of light-operated oxygen production and drug transmission. Compared with the bracket without traditional Chinese medicine panax notoginseng saponins, the proliferation speed of the living microalgae in the hydrogel bracket containing panax notoginseng molecules is higher, so that the overall photosynthesis efficiency of the bracket is obviously improved, more oxygen is generated, and the effect of quickly relieving the local anoxic environment of the wound is achieved. Meanwhile, along with the gradual degradation of the hydrogel scaffold, the microalgae cells and the panax notoginseng drug molecules are gradually released, and the microalgae cells and the panax notoginseng drug molecules provide more bioactive factors for the healing of wounds, promote the migration, proliferation and differentiation of cells to the defect parts, and accelerate the repair and tissue regeneration of the defect parts. The living traditional Chinese medicine scaffold provides a new idea and a new method for developing a novel tissue engineering scaffold material for repairing and treating various chronic tissue defects.
Drawings
FIG. 1 is a schematic view of the in-vivo traditional Chinese medicine stent for wound repair to promote wound healing by releasing oxygen and pseudo-ginseng drug molecules;
FIG. 2 is a scanning electron microscope image of the surface (a) and cross-section (b) of the living traditional Chinese medicine stent of the present invention;
FIG. 3 shows the photosynthetic oxygen production effect of the living body traditional Chinese medicine stent of the present invention;
FIG. 4 shows that skin cells can survive, adhere and spread on the surface of a living traditional Chinese medicine stent (a), and the living traditional Chinese medicine stent has a better effect of promoting cell proliferation (b) compared with a common stent;
fig. 5 is a photograph showing the results of quantitative analysis of the number (b) of hypoxic cells, the number (c) of hypoxic cells, and the survival rate (c) of cells after the in vivo traditional Chinese medicine scaffold and the hypoxic cells are co-cultured for three days, wherein the in vivo traditional Chinese medicine scaffold can effectively relieve the hypoxic state of the cells and improve the survival rate of the cells;
FIG. 6 shows that the living traditional Chinese medicine scaffold can improve the healing speed of the skin wound of a diabetic mouse (a) and accelerate the collagen deposition of new skin (b).
Detailed Description
A living traditional Chinese medicine bracket for wound repair prepared by a microfluidic 3D printing mode is shown in figure 1, and the specific preparation process is as follows:
(1) Preparing an inner phase solution and an outer phase solution:
1.1 Preparing an internal phase solution: the inner phase solution consists of a mixed solution of calcium chloride and polyethylene glycol, and a certain amount of calcium chloride and polyethylene glycol powder are weighed and dissolved in ultrapure water to form a mixed solution of 0.8w/v% calcium chloride and 5w/v% polyethylene glycol as the inner phase solution.
1.2 Preparing an external phase solution: dispersing the panax notoginseng saponins powder and the living microalgae in a mixed dispersion liquid of sodium alginate with the concentration of 2.5w/v% and GelMA with the concentration of 5 w/v%; firstly, taking out the living microalgae cell sap from a refrigerator at 4 ℃, re-warming for 2 hours at room temperature, centrifuging to remove the old culture solution, dispersing the living microalgae cells in the new culture solution, and counting the concentration of the living microalgae cells. Weighing a certain mass of panax notoginseng saponins powder, sodium alginate and GelMA solid from a living body microalgae cell sap with a certain concentration, dissolving the weighed panax notoginseng saponins powder, sodium alginate and GelMA solid in the living body microalgae cell sap to obtain mixed dispersion liquid with the concentrations of the panax notoginseng saponins, the sodium alginate and the GelMA of 25 mu g/mL, 2.5w/v% and 5w/v%, respectively; stirring at 37 ℃, adding a certain mass of a photoinitiator phenyl-2,4,6-trimethylbenzoylimido Lithium (LAP) after the solution is completely dissolved, wherein the final concentration of the photoinitiator is 0.1w/v%.
(2) Assembling the coaxial nested microfluidic chip: drawing two glass capillaries with different sizes by using a microelectrode drawing instrument or an acetylene blowtorch, wherein the caliber of the outer phase capillary is between 250 and 1000 mu m, and the caliber of the inner phase capillary is between 100 and 200 mu m; the micro-fluidic chip is assembled by a glass capillary, a glass slide, a sample application needle and quick-drying glue, wherein the glass capillary is assembled by coaxially nesting an outer-phase conical capillary and an inner-phase spindle-shaped outlet capillary.
(3) Preparing a one-dimensional living traditional Chinese medicine hydrogel hollow fiber element:
the inner and outer phase solution is pumped into disposable medical syringe injectors with corresponding specifications and respectively placed on two peristaltic pumps, the injectors and the microfluidic chip are connected through polyethylene pipes, the flow rate of the inner and outer phase is set, and the peristaltic pumps are started to work; in the microfluidic channel, when the inner phase solution and the outer phase solution meet, the calcium ions and the alginic acid are rapidly crosslinked and cured, hollow fibers with hollow structures are extruded out of an outer phase capillary tube and then collected in a calcium chloride solution with the concentration of 2w/v%, the curing is further enhanced, and simultaneously, the GelMA molecules in the hollow fiber elements are completely photocrosslinked by using ultraviolet light for 5-10 minutes.
(4) Preparing a three-dimensional living traditional Chinese medicine scaffold:
integrating a micro-fluidic chip to a 3D printing platform, controlling the moving position of a micro-fluidic spray head by using a computer program, adjusting the moving speed of the spray head to be 2-15 mm/s, matching the extrusion speed of fibers, and stacking and continuously extruding hollow fiber elements layer by layer on the surface of a clean flat container to form a living traditional Chinese medicine hydrogel fiber support with a three-dimensional communicated porous structure; and after printing is finished, exposing the support to ultraviolet laser irradiation for 5-10 minutes, and performing double crosslinking and curing to obtain the three-dimensional living traditional Chinese medicine support with good mechanical strength and a stable three-dimensional communicated porous structure.
The appearance representation of the three-dimensional living traditional Chinese medicine bracket: after the printing of the stent is finished, observing the distribution condition of the living microalgae in the three-dimensional living traditional Chinese medicine stent under an optical microscope; after the scaffold is freeze-dried, the scaffold is observed by adopting a scanning electron microscope, as shown in figure 2, a porous microporous structure communicated with the scaffold can be obviously seen, and each strut unit is provided with an obvious hollow channel.
Evaluation experiment of photosynthesis oxygen production effect of three-dimensional living body traditional Chinese medicine support:
in order to explore the photosynthetic oxygen production capacity of the live microalgae scaffold, the scaffold is placed under LED light, the distance between the two is kept at 10 cm, the temperature environment is controlled at 25 ℃, and the dissolved oxygen content in the scaffold culture solution is monitored in real time by using a dissolved oxygen micro-sensing electrode. And (3) extracting the chloroplast content at different culture time points by using 95% ethanol to detect the proliferation activity of the microalgae cells. As shown in fig. 3, compared to the stent group without traditional Chinese medicine molecules, the proliferation rate of microalgae cells inside the living traditional Chinese medicine stent is higher, and the overall photosynthetic oxygen production efficiency is higher.
In-vitro tissue regeneration activity evaluation experiment of the three-dimensional living traditional Chinese medicine scaffold:
(1) Cell adhesion, proliferation and migration experiments under normal culture conditions:
1.1 Cell adhesion: adopting vascular endothelial cells as model cells, and culturing in normal oxygen environment. Placing a living traditional Chinese medicine bracket in a 24-well plate, then inoculating cells to the surface of the bracket at the density of 10 ten thousand cells per hole, performing living and dead staining treatment on the cells after 24 hours, and observing and photographing by using a laser confocal microscope. The experimental result is shown in fig. 4a, the living traditional Chinese medicine stent has good cell compatibility, all cells are living cells, and the cells can be adhered and spread on the surface of the stent.
1.2 Cell proliferation: adopting vascular endothelial cells as model cells, and culturing in normal oxygen environment. Cells were seeded at a density of 1 ten thousand cells per well in 24-well plates and after 12 hours the Transwell chamber loaded with the live traditional Chinese medicine scaffold was transferred to the plate for co-culture with the cells. After 1, 3, and 5 days of culture, the cell proliferation was evaluated by measuring the absorbance at 450nm by the CCK8 method (FIG. 4 b), and the cells were stained for dying and observed by confocal laser microscopy.
1.3 Cell migration: 10 ten thousand vascular endothelial cells were seeded in 24-well plates and cultured under hypoxic conditions. After 12 hours a scratch was made with a 200 microliter tip, and the Transwell chamber loaded with the live drug scaffold was then transferred to a well plate for co-culture with the cells. The change condition of the scratch is recorded by photographing at a specific time point, and the reduction rate of the area of the scratch is quantitatively analyzed, so that the traditional Chinese medicine support for the living body has the effect of promoting cell migration.
(2) Evaluation of the activity of the cells in hypoxic culture conditions:
2.1 Adopting vascular endothelial cells as model cells, and placing the model cells in 1%O to simulate the hypoxia condition of wound surface in vivo 2 Is cultured in an anoxic environment. Cells were seeded at a density of 1 ten thousand cells per well in 24-well plates and 12 hours later Transwell chambers loaded with live drug scaffolds were transferred into the plates for co-culture with the cells. The light set was illuminated with an LED bulb with a light intensity of 6000lux for 6 hours per day. After 3 days of anoxic culture, an anoxic fluorescent probe [ Ru (dpp) was used 3 ]Cl 2 Cells were stained (fig. 5 a) and assessed for hypoxia by fluorescent semi-quantitative analysis, counting hypoxic cells (fig. 5 b). The living traditional Chinese medicine bracket can generate oxygen through photosynthesis, so that the cell hypoxia condition is relieved.
2.2 Cell viability assay: placing vascular endothelial cells in 1%O 2 Is cultured in an anoxic environment. Cells were seeded at a density of 1 ten thousand cells per well in 24-well plates and after 12 hours the Transwell chamber loaded with the live traditional Chinese medicine scaffold was transferred to the plate for co-culture with the cells. The light set was illuminated with an LED bulb with a light intensity of 6000lux for 6 hours per day. After 3 days of anaerobic culture, the survival of the cells was evaluated by measuring the absorbance at 450nm by the CCK8 method (FIG. 5 c). And the living cells are dyed and dead, and the laser confocal microscope is adopted for observing and taking pictures, so that the chlorella in the living traditional Chinese medicine bracket can generate oxygen through photosynthesis, the cell hypoxia condition is relieved, and the survival rate of the cells is effectively improved.
In vivo chronic wound repair performance evaluation experiment of the traditional Chinese medicine bracket: a diabetic mouse model was first induced by intraperitoneal injection of Streptozotocin (STZ). 4 weeks after STZ injection, blood glucose was measured in mice and diabetic mice with blood glucose levels above 20mmol/L were selected for subsequent experiments. The groups were randomly divided into five groups: the Chinese medicinal composition comprises (1) a blank control group, (2) a common stent group, (3) a Chinese medicinal stent group, (4) a living stent group and (5) a living Chinese medicinal stent group. A circular full cortical wound of 8 mm diameter was made on the back of each mouse, and a cylindrical scaffold of 8 mm diameter and 2 mm height was implanted into the defect site and subsequently covered with a transparent dressing. All scaffolds were illuminated under LED lights for 2 hours per day. The skin wound healing progress was recorded using a digital camera and the wound healing speed was calculated (fig. 6 a). Two weeks later all mice were sacrificed and sampled for histological analysis, including hematoxylin-eosin staining and masson trichrome staining, as well as immunohistochemical staining for CD31, HIF-1 α and CD163, etc., to assess the microscopic morphology of the neogenetic skin tissue, collagen deposition (fig. 6 b), internal vascularity, tissue hypoxia and inflammatory response, etc.
The above is only a preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above-mentioned embodiments, and all technical solutions belonging to the idea of the present invention belong to the protection scope of the present invention. It should be noted that modifications and embellishments within the scope of the invention may be made by those skilled in the art without departing from the principle of the invention.
Claims (10)
1. A living traditional Chinese medicine bracket for wound repair and a preparation method thereof are characterized by comprising the following steps:
(1) Constructing a one-dimensional living traditional Chinese medicine hydrogel hollow fiber element:
designing and constructing a nested incident channel microfluidic device, taking a water phase solution of a cross-linking agent as an internal phase and a mixed dispersion liquid containing living microalgae and traditional Chinese medicine molecules as an external phase, extruding hollow fibers in the air by using the microfluidic device, and obtaining living traditional Chinese medicine hydrogel hollow fiber elements after cross-linking and curing;
(2) Preparing a three-dimensional living traditional Chinese medicine bracket by microfluidic bioprinting:
directly printing the hollow fiber elements prepared in the step (1) on a flat plate in a 3D printing mode to form a three-dimensional living traditional Chinese medicine hydrogel fiber support which is stacked layer by layer and arranged in a cross mode and has a three-dimensional communicated porous structure, and then performing cross-linking solidification to obtain a three-dimensional living traditional Chinese medicine support;
the size of the channel of the hollow fiber is regulated and controlled by adjusting the diameter of the microfluidic channel and the flow rates of the internal phase fluid and the external phase fluid;
regulating and controlling the bioactivity of the hollow fiber by changing the composition ratio of the living microalgae and the traditional Chinese medicine molecules in the mixed dispersion liquid;
the porous structure, the macroscopic appearance and the size of the three-dimensional living traditional Chinese medicine bracket are regulated and controlled by adjusting the 3D printing program.
2. The in-vivo traditional Chinese medicine stent for wound repair and the preparation method thereof according to claim 1 are characterized in that,
in the step (1), the nested incident channel microfluidic device is formed by assembling a glass capillary tube, a glass slide, a sample application needle head and quick-drying glue; wherein, the glass capillary is assembled by coaxially nesting an external phase capillary and an internal phase capillary.
3. A living body traditional Chinese medicine stent for wound repair and a preparation method thereof according to claim 2, wherein the diameter of the external phase capillary is 250 to 1000 μm, and the diameter of the internal phase capillary is 100 to 200 μm.
4. The in-vivo traditional Chinese medicine stent for wound repair and the preparation method thereof according to claim 1 are characterized in that,
in the step (1), the inner phase is a mixed solution of calcium chloride and polyethylene glycol, and the outer phase is a mixed dispersion of living microalgae, traditional Chinese medicine molecules, sodium alginate and methacrylic acid anhydrized gelatin; the living microalgae is Chlorella, and the Chinese medicinal molecules are Panax notoginsenosides.
5. A living body traditional Chinese medicine stent for wound repair and a preparation method thereof according to claim 4, characterized in that in the mixed dispersion liquid, the concentration of chlorella is 0-1000 ten thousand/mL, the concentration of panax notoginseng saponins is 0-2000 μ g/mL, the concentration of sodium alginate is 0.5-2.5 w/v%, and the concentration of methacrylic anhydrized gelatin is 1-20 w/v%.
6. A living body traditional Chinese medicine bracket for wound repair and a preparation method thereof according to claim 4, characterized in that, in the mixed solution, the concentration of calcium chloride is 0-4 w/v%, and the concentration of polyethylene glycol is 1-50 w/v%.
7. The in-vivo traditional Chinese medicine stent for wound repair and the preparation method thereof according to claim 1 are characterized in that,
in the step (1), the inner diameter of the hollow fiber element is 0-200 μm, the outer diameter is 200-1500 μm, and the aperture size of the three-dimensional living traditional Chinese medicine bracket is 0-5000 μm.
8. The in-vivo traditional Chinese medicine stent for wound repair and the preparation method thereof according to claim 1 are characterized in that,
in the step (2), the hollow fiber elements are stacked in an orthogonal, oblique or parallel manner; the three-dimensional living traditional Chinese medicine bracket is in the shape of a cylinder, a cube or a composite polyhedron.
9. The in-vivo traditional Chinese medicine stent for wound repair and the preparation method thereof according to claim 1 are characterized in that,
in the steps (1) and (2), the crosslinking curing mode is ultraviolet irradiation crosslinking curing.
10. A living body Chinese medicinal stent obtained by the preparation method according to any one of claims 1 to 9.
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Application publication date: 20230307 |