Injection tissue filling material capable of providing cell energy and preparation method thereof

Abstract

The invention belongs to the technical field of biomedical materials, and particularly relates to an injection tissue filling material capable of providing cell energy and a preparation method thereof, the injection tissue filling material comprises a gel carrier and magnesium-containing microspheres uniformly distributed in the gel carrier; the magnesium-containing microsphere is obtained by emulsifying a degradable polyester high polymer material and a magnesium-containing compound, and the magnesium-containing compound is uniformly distributed on the surface and inside the magnesium-containing microsphere; wherein the magnesium-containing microspheres account for 5-50% of the mass of the tissue filling material; the mass ratio of the gel carrier to the tissue filling material is 0.5-10%; the concentration of magnesium ions in the magnesium-containing microspheres is 30-120 [mu] g / g; the magnesium-containing degradable polymer microspheres are prepared through emulsification and are mixed in the gel carrier to obtain the injectable composite filling material, so that the situation that magnesium ions are released too quickly and are greatly lost due to local direct use of a magnesium compound is avoided, and the effect of the tissue filling material for improving cell energy is ensured.

Description

Technical Field

[0001] The present invention belongs to the technical field of biomedical materials, and in particular relates to an injectable tissue filling material capable of providing cell energy and a preparation method thereof. Background Art

[0002] In recent years, research in the fields of skin tissue and bone tissue remains a hot topic in the field of tissue engineering. Using the three major elements of tissue engineering to construct materials with activity and regenerative capabilities is a method that has been studied more frequently and has relatively mature technology in this field. Based on previous studies, more and more researchers have gradually begun to focus on how to develop materials and treatments that can accelerate the speed of tissue repair based on the mechanism of the body's own repair process. Among them, the study of cellular energy has become a new hot topic. The dynamic changes in cellular energy metabolism are the core driving force of tissue repair. Skin repair relies more on rapid energy conversion to adapt to changes in the surface environment, while bone repair requires coordinated multi-stage metabolic reprogramming (glycolysis-OXPHOS conversion). Generally speaking, cellular energy provides power for key links such as cell proliferation, migration, differentiation and matrix synthesis. Increasing the level of cellular energy to a certain extent can effectively accelerate the process of tissue repair.

[0003] Currently, much research on cellular energy is focused on substances in the tricarboxylic acid cycle (TCA). The TCA cycle is a crucial step in the mitochondrial ATP production process. Cellular respiration and ATP production can be significantly accelerated by exogenous additions of succinate, fumarate, malate, and other dicarboxylic acids. Exogenous addition of any of these dicarboxylic acids results in significant oxygen consumption, carbon dioxide production, and ATP production. Furthermore, studies have shown that the addition of succinate, fumarate, malate, and other key intermediates in the TCA cycle significantly increases the rate of the TCA cycle, thereby promoting ATP production. ATP is divided into intracellular ATP (endogenous ATP) and extracellular ATP (exogenous ATP), which play roles in energy supply, signal transduction, and protein activation, respectively. Within the cell, almost all forms of energy are directly converted from ATP and utilized by the cell, including mechanical energy, thermal energy, chemical potential energy, and electrical potential energy. ATP directly participates in various biochemical reactions within the cell.

[0004] Based on the above mechanism, Zhang et al. applied for an invention patent (CN 102936407 B) and prepared an energy biomaterial based on the tricarboxylic acid cycle. This material is obtained by polymerizing monomer materials in the tricarboxylic acid cycle, such as succinic acid, citric acid, isocitric acid, fumaric acid, malic acid, etc., with diols and their derivatives to obtain a variety of polymer materials. Substances released during the degradation of the polymers in the body enter cells and participate in the tricarboxylic acid cycle, accelerating the generation of ATP in cells, and promoting cell growth, proliferation and tissue repair.

[0005] However, in this method, the release of energy substances depends on the degradation of polymers in the body. It cannot be guaranteed that the degradation products will enter the cells completely as expected monomer degradation products during the degradation process. For example, in the copolymer of citric acid and ethylene glycol, during the degradation process, will there be a molecule in the form of a citric acid monomer and two ethylene glycol monomers connected to enter the cells? Whether this form can exert the above-mentioned effect of increasing cell energy levels after entering the cells has not been verified.

[0006] Therefore, how to overcome the defect of uncontrollable cellular energy in current injectable tissue filling materials is a technical problem that urgently needs to be solved in this field.

[0007] It should be noted that the above information disclosed in this background technology section is only used to understand the background technology of the present application concept, and therefore, the above description is not considered to constitute information of the prior art. Summary of the Invention

[0008] The embodiments of the present disclosure at least provide an injectable tissue filling material that can provide cellular energy and a preparation method thereof.

[0009] In a first aspect, embodiments of the present disclosure provide an injectable tissue filler material capable of providing cellular energy, comprising: a gel carrier and magnesium-containing microspheres uniformly distributed therein; the magnesium-containing microspheres are obtained by emulsifying a degradable polyester polymer material with a magnesium-containing compound, and the magnesium-containing compound is uniformly distributed on the surface and inside of the magnesium-containing microspheres; the magnesium-containing microspheres account for 5 to 50% by mass of the tissue filler material; the gel carrier accounts for 0.5 to 10% by mass of the tissue filler material; and the concentration of magnesium ions in the magnesium-containing microspheres is 30 to 120 μg / g.

[0010] In an optional embodiment, the magnesium-containing compound includes any one or more combinations of magnesium oxide, magnesium carbonate, and magnesium hydroxide.

[0011] In an optional embodiment, the particle size of the magnesium-containing microspheres ranges from 10 to 200 μm; the degradable polyester polymer material includes polylactic acid, polycaprolactone, polydioxanone, polylactic acid-glycolic acid copolymer, and polyhydroxyalkanoate.

[0012] In an optional embodiment, the gel carrier includes any one of a high-viscosity non-cross-linked solution and a cross-linked gel; the material of the gel carrier includes any one or more combinations of sodium hyaluronate, collagen, polyvinyl alcohol, polyethylene glycol, sodium carboxymethyl cellulose, sodium alginate, dextran, chitosan, and gelatin; and the molecular weight of the material of the gel carrier is 50,000 to 3,000,000; and the dynamic viscosity of the gel carrier is 50,000 to 500,000 mPa·s.

[0013] In an optional embodiment, it further comprises: any one or more of bioactive energy substances, water-soluble magnesium-containing substances and bioactive factors.

[0014] In an optional embodiment, the bioactive energy substance includes any one or more combinations of citric acid, aconitic acid, isocitric acid, oxalosuccinic acid, α-ketoglutaric acid, succinyl coenzyme A, succinic acid, fumaric acid, malic acid, and adenosine triphosphate.

[0015] In an optional embodiment, the water-soluble magnesium-containing substance includes any one or more combinations of magnesium chloride, magnesium malate, and magnesium citrate.

[0016] In an optional embodiment, the bioactive factor includes any one of bone morphogenetic protein and epidermal growth factor or a combination of both.

[0017] In a second aspect, the embodiments of the present disclosure further provide a method for preparing an injectable tissue filler material that can provide cellular energy as described above, comprising the following steps: weighing a degradable polyester polymer material, stirring it thoroughly until dissolved, adding a magnesium-containing compound, and ultrasonically dispersing it to obtain a magnesium-containing polymer solution; preparing a PVA solution, continuously stirring it and adding the magnesium-containing polymer solution for emulsification, heating it to a high temperature, and freeze-drying it to obtain dried magnesium-containing microspheres; weighing the raw materials to prepare a gel carrier; and mixing the magnesium-containing microspheres with the gel carrier to obtain an injectable tissue filler material that can provide cellular energy.

[0018] In an optional embodiment, the emulsification reaction temperature is not higher than 60° C., and the reaction time is not more than 1 h.

[0019] The beneficial effects of the present invention are that the injectable tissue filling material that can provide cellular energy and the preparation method thereof prepare magnesium-containing degradable polymer microspheres by emulsification and mix them in a gel carrier to obtain an injectable composite filling material. The magnesium-containing compound contained in the microspheres is released in the in vivo environment as the microspheres degrade, and the release rate is adjustable, thereby avoiding the excessive release and large-scale loss of magnesium ions caused by the direct local use of the magnesium compound, thereby ensuring the effect of the tissue filling material in enhancing cellular energy.

[0020] Other features and advantages of the present invention will be described in the following description, and in part will become apparent from the description, or understood by practicing the present invention. The purpose and other advantages of the present invention are realized and obtained by the structures particularly pointed out in the description and the drawings.

[0021] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, preferred embodiments are given below and described in detail with reference to the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the specific embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0023] Figure 1 This is a microscopic morphology of the magnesium-containing microspheres prepared in Example 2 provided in the embodiments of the present disclosure;

[0024] Figure 2 This is a microscopic morphology of the magnesium-containing microspheres prepared in Example 3 provided in the embodiments of the present disclosure;

[0025] Figure 3 This is a scanning electron microscope morphology image of the magnesium-containing microspheres prepared in Example 1 provided in the embodiments of the present disclosure;

[0026] Figure 4 This is a particle size distribution diagram of the magnesium-containing microspheres prepared in Example 1 provided in the embodiments of the present disclosure;

[0027] Figure 5 The cell compatibility results of the magnesium-containing microspheres prepared in Examples 1-4 provided in the present disclosure;

[0028] Figure 6 The bulk density test results of the magnesium-containing microspheres prepared in Example 5 provided in the embodiments of the present disclosure;

[0029] Figure 7 The ATP content test results of the magnesium-containing microspheres prepared in Example 1 provided in the embodiments of the present disclosure;

[0030] Figure 8 H&E tissue section data of mice implanted subcutaneously for one month without magnesium microspheres in Comparative Example 1 provided in the embodiments of the present disclosure;

[0031] Figure 9 H&E tissue section data of mice implanted subcutaneously with magnesium microspheres for one month in Example 1 provided in the embodiments of the present disclosure;

[0032] Figure 10 This is the Masson tissue section data of mice implanted subcutaneously for one month without magnesium microspheres in Comparative Example 1 provided in the embodiments of the present disclosure;

[0033] Figure 11 The data of Masson tissue sections of mice implanted subcutaneously with magnesium microspheres for one month in Example 1 provided in the embodiments of the present disclosure are as follows: DETAILED DESCRIPTION

[0034] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. All other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making any creative efforts shall fall within the scope of protection of the present invention.

[0035] As used herein, the phrases "in one embodiment," "according to one embodiment," "in some embodiments," and the like generally refer to the fact that the particular feature, structure, or characteristic following the phrase may be included in at least one embodiment of the present disclosure. Thus, a particular feature, structure, or characteristic may be included in more than one embodiment of the present disclosure, such that these phrases do not necessarily refer to the same embodiment. As used herein, the terms "example," "exemplary," and the like are used to "serve as an example, instance, or illustration." Any implementation, aspect, or design described herein as "example" or "exemplary" is not necessarily to be construed as preferred or advantageous over other implementations, aspects, or designs. Instead, the use of the terms "example," "exemplary," and the like is intended to present concepts in a concrete manner.

[0036] Herein, example embodiments of the present disclosure will be described in more detail with reference to the accompanying drawings. As used herein, expressions such as "at least one of..." when following a list of elements modify the entire list of elements, rather than modifying individual elements in the list. For example, the expression "at least one of a, b, and c" should be understood to include only a, only b, only c, both a and b, both a and c, both b and c, or all of a, b, and c.

[0037] The terms used herein are only used to describe specific exemplary configurations and are not intended to be limiting. As used herein, the singular articles "a", "an" and "the" may also be intended to include plural forms, unless otherwise clearly indicated herein. The terms "comprise", "include" and "have" are inclusive and therefore specify the presence of features, steps, operations, elements and / or components, but do not exclude the presence or addition of one or more other features, steps, operations, elements, components and / or combinations thereof. The method steps, processes and operations described herein should not be interpreted as necessarily requiring them to be performed in the particular order discussed or shown, unless specifically identified as an execution order. Additional or alternative steps may be adopted.

[0038] The following embodiments of the present invention are described in detail with reference to the accompanying drawings. In the absence of conflict, the following embodiments and features therein may be combined with each other.

[0039] The present disclosure provides an injectable tissue filler material capable of providing cellular energy, comprising: a gel carrier and magnesium-containing microspheres uniformly distributed therein; the magnesium-containing microspheres are obtained by emulsifying a degradable polyester polymer material with a magnesium-containing compound, and the magnesium-containing compound is uniformly distributed on the surface and inside of the magnesium-containing microspheres; the magnesium-containing microspheres account for 5 to 50% by mass of the tissue filler material; the gel carrier accounts for 0.5 to 10% by mass of the tissue filler material; and the magnesium ion concentration in the magnesium-containing microspheres is 30 to 120 μg / g.

[0040] In some embodiments, specifically, the magnesium-containing compound includes any one or more combinations of magnesium oxide, magnesium carbonate, and magnesium hydroxide.

[0041] Specifically, magnesium ions (Mg 2+ ) is one of the most abundant divalent cations in cells. As an essential cofactor of ATP and an activator of various enzymes, it is directly involved in the regulation of cellular energy metabolism. Its role runs through key links such as ATP synthesis, transfer, utilization, and mitochondrial function, and it plays an irreplaceable role in the repair of skin and bone tissue. Under physiological conditions, ATP must react with Mg. 2+ Bind (to form MgATP 2- ) can maintain stability and function. Free ATP is easily hydrolyzed, while Mg 2+ By shielding the negative charge of the phosphate group, the energy barrier of ATP hydrolysis is lowered, making it a "universal currency" for energy transfer. About 95% of ATP in cells is converted to Mg. 2+ In addition, magnesium ions have multiple effects on energy metabolism pathways, such as: (1) glycolysis, hexokinase and phosphofructokinase-1 (PFK-1) are dependent on Mg. 2+ Activation promotes the decomposition of glucose into pyruvate. When pyruvate kinase converts phosphoenolpyruvate (PEP) into pyruvate, Mg is required. 2+ Participate in the production of ATP; (2) tricarboxylic acid cycle (TCA cycle), dehydrogenase complexes (such as α-ketoglutarate dehydrogenase) require Mg 2+ As a cofactor, it promotes the generation of NADH and FADH2, providing energy for the electron transport chain. The activity of citrate synthase is affected by Mg. 2+ Regulation, affecting TCA cycle flux; (3) Oxidative phosphorylation (OXPHOS), the rotational catalytic mechanism of ATP synthase (complex V) depends on Mg 2+ Stabilizes the conformation of its F1 subunit and promotes the condensation of ADP and Pi into ATP. Magnesium deficiency will lead to a decrease in mitochondrial membrane potential and a decrease in the efficiency of the electron transport chain. (4) Fatty acid oxidation, carnitine palmitoyltransferase (CPT-1) requires Mg 2+ Activation, promoting the entry of long-chain fatty acids into mitochondria for β-oxidation.

[0042] Specifically, magnesium ions play an important role in the repair of skin and bone tissue. 2+ By activating the ERK / MAPK pathway, it promotes collagen synthesis (requires a large amount of ATP); in terms of bone tissue repair, Mg 2+ Promotes MSCs differentiation into osteoblasts by upregulating Runx2 expression, increasing ALP activity and mineralized nodule formation (requires continuous ATP supply), Mg 2+ When it is deficient, osteoclast activity is enhanced, bone resorption is excessive, the repair balance is disrupted, and Mg is released during degradation. 2+ , locally improve ATP synthesis efficiency and accelerate bone healing.

[0043] In some embodiments, specifically, the particle size of the magnesium-containing microspheres ranges from 10 to 200 μm; the degradable polyester polymer material includes polylactic acid, polycaprolactone, polydioxanone, polylactic acid-glycolic acid copolymer, and polyhydroxyalkanoate.

[0044] In some embodiments, specifically, the gel carrier includes any one of a high-viscosity non-cross-linked solution and a cross-linked gel; the material of the gel carrier includes any one or more combinations of sodium hyaluronate, collagen, polyvinyl alcohol, polyethylene glycol, sodium carboxymethyl cellulose, sodium alginate, dextran, chitosan, and gelatin; and the molecular weight of the material of the gel carrier is 50,000 to 3,000,000; the dynamic viscosity of the gel carrier is 50,000 to 500,000 mPa·s.

[0045] In some embodiments, specifically, it further comprises: any one or more of bioactive energy substances, water-soluble magnesium-containing substances and bioactive factors.

[0046] In some embodiments, specifically, the bioactive energy substance includes any one or more combinations of citric acid, aconitic acid, isocitric acid, oxalosuccinic acid, α-ketoglutaric acid, succinyl-CoA, succinic acid, fumaric acid, malic acid, and adenosine triphosphate.

[0047] In some embodiments, specifically, the water-soluble magnesium-containing substance includes any one or more combinations of magnesium chloride, magnesium malate, and magnesium citrate.

[0048] In some embodiments, specifically, the bioactive factor includes any one or a combination of two of bone morphogenetic protein and epidermal growth factor.

[0049] The present disclosure also provides a method for preparing the aforementioned injectable tissue filler material capable of providing cellular energy, comprising the following steps: weighing a degradable polyester polymer material, stirring it thoroughly until dissolved, adding a magnesium-containing compound, and ultrasonically dispersing it to obtain a magnesium-containing polymer solution; preparing a PVA solution, continuously stirring it, and adding the magnesium-containing polymer solution for emulsification, heating it to a high temperature, and freeze-drying it to obtain dried magnesium-containing microspheres; weighing the raw materials to prepare a gel carrier; and mixing the magnesium-containing microspheres with the gel carrier to obtain the injectable tissue filler material capable of providing cellular energy.

[0050] Specifically, the injectable tissue filling material that can provide cellular energy can be applied to the fields of skin defect repair, skin volume enhancement, bone defect repair, etc.

[0051] Comparative Example 1

[0052] (1) Weigh 10 g of PLGA (polylactic acid-co-glycolic acid) raw material, add dichloromethane to prepare a 3% solution, and stir thoroughly until the PLGA is completely dissolved; prepare 5 L of 1% PVA (polyvinyl alcohol) solution, set the speed to 500 rpm and stir continuously, add PLGA solution during stirring for emulsification, emulsify for 30 minutes, heat to 45°C, and continue for 24 hours to remove the dichloromethane solvent, then use a sieve to wash and collect to obtain wet PLGA microspheres, and after freeze-drying, obtain dry PLGA microspheres, and sieve the PLGA microspheres to obtain microspheres with an average particle size of 55 μm;

[0053] (2) Sodium hyaluronate with a molecular weight of 1.5 million was used as a raw material and BDDE (butylene glycol glycidyl ether) was used as a cross-linking agent to prepare a cross-linked gel with an average particle size of 500 μm and a dynamic viscosity of 350,000 mPa·s;

[0054] (3) Mixing PLGA microspheres with cross-linked sodium hyaluronate gel and injecting the mixture into a pre-filled syringe to obtain an injectable tissue filling material containing PLGA microspheres.

[0055] Example 1

[0056] (1) Weigh 10 g of PLA (polylactic acid) raw material, add chloroform to prepare a 3% solution, and stir thoroughly until PLA is completely dissolved; add 400 mg of magnesium oxide and 400 mg of magnesium carbonate, and ultrasonically disperse the two magnesium-containing substances in the PLA solution; prepare 5 L of 1% PVA solution, set the speed to 900 rpm and stir continuously, add PLA solution during stirring for emulsification, emulsify after 30 minutes, heat to 48 ° C, and continue for 24 hours to remove the chloroform solvent, then use a sieve to wash and collect to obtain wet PLA-Mg microspheres, and after freeze-drying, obtain dry PLA-Mg microspheres, and sieve the PLA-Mg microspheres to obtain microspheres with an average particle size of 55 μm; the magnesium ion concentration in the microspheres is measured to be 40 μg / g;

[0057] (2) Sodium hyaluronate with a molecular weight of 900,000 was selected as a raw material and cystamine was used as a cross-linking agent to prepare a cross-linked gel with an average particle size of 800 μm and a dynamic viscosity of 450,000 mPa·s;

[0058] (3) PLA-Mg microspheres were mixed with cross-linked sodium hyaluronate gel, with the mass ratio of PLA-Mg microspheres being 10% and the mass ratio of gel being 5%, and the mixture was poured into a pre-filled syringe to obtain an injectable tissue filling material containing PLA-Mg microspheres.

[0059] Example 2

[0060] (1) Weigh 15 g of PLGA raw material, add chloroform to prepare a 3% solution, and stir thoroughly until the PLGA is completely dissolved; add 500 mg of magnesium oxide, and ultrasonically treat to evenly disperse the magnesium-containing substance in the PLGA solution; prepare 5 L of 1% PVA solution, set the speed to 800 rpm and stir continuously, add PLGA solution during stirring for emulsification, emulsify for 30 minutes, heat to 45°C, and continue for 24 hours to remove the chloroform solvent, then use a sieve to wash and collect to obtain wet PLGA-Mg microspheres, and after freeze-drying, obtain dry PLGA-Mg microspheres, and sieve the PLGA-Mg microspheres to obtain microspheres with an average particle size of 60 μm; the magnesium ion concentration in the microspheres is measured to be 120 μg / g;

[0061] (2) Sodium hyaluronate with a molecular weight of 50,000 was selected as a raw material and cystamine was used as a cross-linking agent to prepare a cross-linked gel with an average particle size of 1000 μm and a dynamic viscosity of 500,000 mPa·s;

[0062] (3) PLGA-Mg microspheres were mixed with cross-linked sodium hyaluronate gel, with the mass ratio of PLGA-Mg microspheres being 50% and the mass ratio of gel being 10%, and the mixture was poured into a pre-filled syringe to obtain an injectable tissue filling material containing PLGA-Mg microspheres.

[0063] Example 3

[0064] (1) Weigh 10 g of PLGA raw material, add chloroform to prepare a 3% solution, and stir thoroughly until the PLGA is completely dissolved; add 250 mg of magnesium oxide, and ultrasonically treat to evenly disperse the magnesium-containing substance in the PLGA solution; prepare 5 L of 1% PVA solution, set the speed to 300 rpm and stir continuously, add PLGA solution during stirring for emulsification, emulsify for 30 minutes, heat to 45°C, and continue for 24 hours to remove the chloroform solvent, then use a sieve to wash and collect to obtain wet PLGA-Mg microspheres, and after freeze-drying, obtain dry PLGA-Mg microspheres, and sieve the PLGA-Mg microspheres to obtain microspheres with an average particle size of 10 μm; the magnesium ion concentration in the microspheres is measured to be 30 μg / g;

[0065] (2) Polyethylene glycol with a molecular weight of 2 million was selected as the raw material and polylysine was used as the cross-linking agent to prepare a cross-linked gel with an average particle size of 100 μm and a dynamic viscosity of 50,000 mPa·s;

[0066] (3) PLGA-Mg microspheres were mixed with cross-linked polyethylene glycol gel, with the mass ratio of PLGA-Mg microspheres being 15% and the mass ratio of gel being 8%, and the mixture was poured into a pre-filled syringe to obtain an injectable tissue filling material containing PLGA-Mg microspheres.

[0067] Example 4

[0068] (1) Weigh 18 g of PLA raw material, add dichloromethane to prepare a 5% solution, and stir thoroughly until PLA is completely dissolved; add 680 mg of magnesium carbonate, and ultrasonically disperse the magnesium-containing substance in the PLA solution; prepare 5 L of 1% PVA solution, set the speed to 800 rpm and stir continuously, add PLA solution during stirring for emulsification, emulsify for 30 minutes, heat to 45°C, and continue for 24 hours to remove the dichloromethane solvent, then use a sieve to wash and collect to obtain wet PLA-Mg microspheres, and after freeze-drying, obtain dry PLA-Mg microspheres, and sieve the PLA-Mg microspheres to obtain microspheres with an average particle size of 100 μm; the magnesium ion concentration in the microspheres is measured to be 80 μg / g;

[0069] (2) Sodium hyaluronate with a molecular weight of 3 million was selected as the raw material and BDDE was used as the cross-linking agent to prepare a cross-linked gel with an average particle size of 300 μm and a dynamic viscosity of 480,000 mPa·s;

[0070] (3) PLA-Mg microspheres were mixed with cross-linked sodium hyaluronate gel, with the mass ratio of PLA-Mg microspheres being 5% and the mass ratio of gel being 0.5%, and the mixture was poured into a pre-filled syringe to obtain an injectable tissue filling material containing PLA-Mg microspheres.

[0071] Example 5

[0072] (1) Weigh 20 g of PCL raw material, add dichloromethane to prepare a 4% solution, and stir thoroughly until PCL (polycaprolactone) is completely dissolved; add 500 mg of magnesium carbonate and 300 mg of magnesium hydroxide, and ultrasonically disperse the two magnesium-containing substances in the PCL solution; prepare 5 L of 1% PVA solution, set the speed to 800 rpm and stir continuously, add PCL solution during stirring for emulsification, emulsify after 30 minutes, heat to 45 ° C, and continue for 24 hours to remove the dichloromethane solvent, then use a sieve to wash and collect to obtain wet PCL-Mg microspheres, and after freeze-drying, obtain dry PCL-Mg microspheres. The PCL-Mg microspheres are sieved with a sieve to obtain microspheres with an average particle size of 40 μm; the magnesium ion concentration in the microspheres is measured to be 65 μg / g;

[0073] (2) Selecting collagen raw material with a molecular weight of 600,000, preparing it into a non-cross-linked gel with a dynamic viscosity of 100,000 mPa·s;

[0074] (3) PCL-Mg microspheres were mixed with non-cross-linked collagen gel, with a PCL-Mg microsphere mass ratio of 6% and a gel mass ratio of 3%. 50 mg of citric acid and 10 μg of bone morphogenetic protein were added and poured into a prefilled syringe to obtain an injectable tissue filling material containing PCL-Mg microspheres.

[0075] Example 6

[0076] (1) Weigh 10 g of PCL raw material, add dichloromethane to prepare a 3% solution, and stir thoroughly until PCL is completely dissolved; add 300 mg of magnesium carbonate, and ultrasonically treat to evenly disperse the magnesium-containing substance in the PCL solution; prepare 5 L of 1% PVA solution, set the speed to 800 rpm and stir continuously, add PCL solution during stirring for emulsification, emulsify for 30 minutes, heat to 50°C, and continue for 24 hours to remove the dichloromethane solvent, then use a sieve to wash and collect to obtain wet PCL-Mg microspheres, and after freeze-drying, obtain dry PCL-Mg microspheres, and sieve the PCL-Mg microspheres to obtain microspheres with an average particle size of 60 μm; the magnesium ion concentration in the microspheres is measured to be 35 μg / g;

[0077] (2) Selecting collagen raw material with a molecular weight of 800,000 and preparing it into a non-cross-linked gel with a dynamic viscosity of 100,000 mPa·s;

[0078] (3) PCL-Mg microspheres were mixed with non-cross-linked collagen gel, with a mass ratio of PCL-Mg microspheres of 7% and a mass ratio of gel of 2%. 15 μg of epidermal growth factor was added and the mixture was poured into a pre-filled syringe to obtain an injectable tissue filling material containing PCL-Mg microspheres.

[0079] like Figure 1 As shown in the microscopic morphology of the magnesium-containing microspheres prepared in Example 2 of the present invention, the shape is spherical; Figure 2 The microscopic morphology of the magnesium-containing microspheres prepared in Example 3 is spherical in shape. In order to investigate the actual morphology of the microspheres, the microspheres in Example 1 of the present invention were characterized by scanning electron microscopy. Figure 3 As shown in Figure 2, there are some hollow structures on the surface of the microspheres, which is due to the loss of some magnesium-containing compounds after the emulsification process, resulting in holes; Figure 4 As shown, the particle size distribution of the microspheres was characterized. The peak shape in the distribution diagram was single, indicating that the particle size of the microspheres after screening was relatively uniform.

[0080] like Figure 4-6 As shown, after the large-size calcium carbonate particles are compounded with the gel, the stability of the particles can be ensured, and they can be evenly distributed inside the gel, which is convenient for the preparation and processing of the product.

[0081] like Figure 5 As shown in FIG, the microspheres in comparative example 1 and the four examples were subjected to cell compatibility tests. As can be seen from the figure, none of the five groups of microspheres caused significant cytotoxicity, indicating that the microspheres have high biosafety.

[0082] like Figure 6 As shown, the bulk density of the microspheres was tested and compared with that of Comparative Example 1. The bulk density of the magnesium-containing microspheres in Example 5 was much lower than that of the microspheres in Comparative Example 1, indicating that the magnesium-containing compound can significantly reduce the density of the microspheres during the emulsification process, thereby facilitating the release of the magnesium-containing compound during the degradation process of the microspheres.

[0083] like Figure 7 As shown, the ATP levels of different groups were tested. After the addition of magnesium-containing microspheres, the release of magnesium ions promoted the production rate of intracellular ATP, and the ATP content was significantly higher than that of the blank control group and the microsphere group without magnesium in Comparative Example 1.

[0084] like Figure 8-11 The microspheres in Comparative Example 1 and Example 1 were implanted subcutaneously in mice. The data showed that both groups of microspheres did not produce serious toxicity. In addition, due to the low bulk density and porous structure of the magnesium-containing microspheres, it was obvious that cells could enter the microspheres and grow.

[0085] In summary, the injectable tissue filler material that can provide cellular energy and its preparation method prepare magnesium-containing degradable polymer microspheres by emulsification and mix them in a gel carrier to obtain an injectable composite filler material. The magnesium-containing compound contained in the microspheres is released in the in vivo environment as the microspheres degrade, and the release rate is adjustable, avoiding the direct local use of magnesium compounds that would cause excessive release and large-scale loss of magnesium ions, thereby ensuring the effect of the tissue filler material in enhancing cellular energy.

[0086] With the above-described preferred embodiments of the present invention as a guide, and with reference to the above description, relevant personnel are fully capable of making various changes and modifications without departing from the technical scope of this invention. The technical scope of this invention is not limited to the contents of the specification and must be determined according to the scope of the claims.

Claims

1. An injectable tissue filling material that can provide cellular energy, characterized in that: include: A gel carrier and magnesium-containing microspheres uniformly distributed therein; The magnesium-containing microspheres are obtained by emulsifying a degradable polyester polymer material and a magnesium-containing compound, and the magnesium-containing compound is evenly distributed on the surface and inside of the magnesium-containing microspheres; The mass ratio of the magnesium-containing microspheres to the tissue filling material is 5 to 50%; The mass ratio of the gel carrier to the tissue filling material is 0.5 to 10%; The concentration of magnesium ions in the magnesium-containing microspheres is 30 to 120 μg / g.

2. The injectable tissue filling material capable of providing cellular energy according to claim 1, wherein: The magnesium-containing compound includes any one or more combinations of magnesium oxide, magnesium carbonate, and magnesium hydroxide.

3. The injectable tissue filler material capable of providing cellular energy according to claim 1, wherein: The particle size of the magnesium-containing microspheres ranges from 10 to 200 μm; The degradable polyester polymer material includes polylactic acid, polycaprolactone, polydioxanone, polylactic acid-glycolic acid copolymer, and polyhydroxyalkanoate.

4. The injectable tissue filler material capable of providing cellular energy according to claim 1, wherein: The gel carrier includes any one of a high-viscosity non-cross-linked solution and a cross-linked gel; The material of the gel carrier includes any one or more combinations of sodium hyaluronate, collagen, polyvinyl alcohol, polyethylene glycol, sodium carboxymethyl cellulose, sodium alginate, dextran, chitosan, and gelatin; And, the molecular weight of the material of the gel carrier is 50,000 to 3,000,000; The dynamic viscosity of the gel carrier is 50,000 to 500,000 mPa·s.

5. The injectable tissue filling material capable of providing cellular energy according to claim 1, wherein: Also includes: Any one or more of biologically active energy substances, water-soluble magnesium-containing substances and biologically active factors.

6. The injectable tissue filler material capable of providing cellular energy according to claim 5, wherein: The bioactive energy substance includes any one or more combinations of citric acid, aconitic acid, isocitric acid, oxalosuccinic acid, α-ketoglutaric acid, succinyl coenzyme A, succinic acid, fumaric acid, malic acid, and adenosine triphosphate.

7. The injectable tissue filler material capable of providing cellular energy according to claim 5, wherein: The water-soluble magnesium-containing substance includes any one or more combinations of magnesium chloride, magnesium malate, and magnesium citrate.

8. The injectable tissue filler material capable of providing cellular energy according to claim 5, wherein: The bioactive factors include any one of bone morphogenetic protein and epidermal growth factor or a combination of both.

9. A method for preparing an injectable tissue filler material capable of providing cell energy according to any one of claims 1 to 8, characterized in that: The steps include: Weigh a degradable polyester polymer material and stir thoroughly until dissolved, add a magnesium-containing compound, and ultrasonically disperse to obtain a magnesium-containing polymer solution; Prepare a PVA solution, continue stirring and add a magnesium-containing polymer solution for emulsification, heat to a high temperature, and freeze-dry to obtain dry magnesium-containing microspheres; Weighing raw materials to prepare a gel carrier; The magnesium-containing microspheres are mixed with a gel carrier to obtain an injectable tissue filling material that can provide cell energy.

10. The preparation method according to claim 9, characterized in that The emulsification reaction temperature is not higher than 60° C., and the reaction time is not more than 1 hour.

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