Disclosure of Invention
Object of the Invention
In view of the technical problems described in the above prior art section, it is an object of the present invention to provide a chronic healing deep skin wound protective repair composition as well as a method for preparing said composition and a dressing formulation comprising said composition.
Technical proposal
In order to achieve the above purpose, the present invention adopts the following technical scheme:
Scheme 1, a chronic healing deep skin wound protective repair composition, wherein the composition comprises or consists of the following components:
Based on the total weight of the composition:
(1) About 4.5 to about 30.0 wt%, preferably about 10 to about 25wt%, of a phosphosilicate-calcium bioactive glass that is substantially free of alkali metal oxides, such as substantially free of Na2 O; wherein the phosphorus silicon calcium bioactive glass preferably comprises or consists of the following components:
Based on the total weight of the phosphorus silicon calcium bioactive glass:
From about 46 to about 53 wt%, preferably from about 49 to about 52 wt%, more preferably from about 50 to about 51 wt% SiO2,
About 26 to about 43 weight%, preferably about 31 to about 35 weight%, more preferably about 33 to about 34 weight%, of CaO, and
About 5wt% to about 28 wt%, preferably about 14 wt% to about 20 wt%, more preferably about 15 wt% to about 17 wt%, and most preferably about 16 wt% P2O5;
(2) About 2 to about 10 weight percent, preferably about 4 to about 8 weight percent, of hyaluronic acid and/or a salt thereof, preferably sodium salt of hyaluronic acid, wherein the hyaluronic acid and/or salt thereof preferably has a molecular weight in the range of about 200 to about 1200 KDa (kilodaltons), more preferably in the range of about 400 to about 800 KDa; and
(3) From about 60 to about 92.5 wt%, preferably from about 67 to about 86 wt%, of an excipient and/or a slow release agent, wherein the excipient and/or slow release agent preferably has a cone penetration in the range of from about 160 to about 190, preferably from about 170 to about 185.
Scheme 2 the chronic healing deep skin wound protective repair composition according to scheme 1 above wherein the excipient and/or slow release agent comprises one or more of petrolatum and lanolin, preferably in a weight ratio of about (0-18): 100, preferably about (2-10): 100 lanolin and petrolatum.
Scheme 3 the chronic healing deep skin wound protective repair composition according to any one of the above schemes 1 to 2, wherein the phosphosilicate calcium bioactive glass contained in the composition has the form of a particulate porous material having a specific surface area of not less than about 200 m2/g, preferably in the range of about 290 to about 350 m2/g, and having a particle size in the range of about 10 to about 2000 μm, preferably in the range of about 100 to about 180 μm.
Scheme 4 the chronic healing deep skin wound protective repair composition according to any one of the above schemes 1 to 3, wherein the hyaluronic acid and/or salt thereof contained in the composition has the form of a particulate material having a particle size of not more than about 220 μm, preferably in the range of about 70 to about 180 μm.
Scheme 5 the chronic healing deep skin wound protective repair composition according to any one of the above schemes 1 to 3, wherein the phosphosilicate calcium bioactive glass and the hyaluronic acid and/or its salt are present in the composition in the form of a composite particulate material, the hyaluronic acid and/or its salt being coated on the outer layer of the particulate phosphosilicate calcium bioactive glass material.
Scheme 6, a method for preparing a chronic healing deep skin wound protection repair composition according to any one of the above schemes 1 to 4, wherein the method comprises the steps of:
s1: liquefying the excipient and/or the sustained release agent, and
S2: dispersing the phosphorus silicon calcium bioactive glass and the hyaluronic acid and/or the salt thereof in the liquefied excipient and/or the sustained release agent obtained in the step S1 sequentially or simultaneously according to any sequence, thereby obtaining the chronic healing deep skin wound protection and repair composition.
Scheme 7, a method for preparing a chronic healing deep skin wound protection repair composition according to scheme 5 above, wherein the method comprises the steps of:
S1: liquefying the excipient and/or the sustained release agent,
S2: dispersing the composite particulate material in the liquefied excipient and/or slow release agent obtained from step S1, thereby obtaining the chronic healing deep skin wound protective repair composition, wherein the composite particulate material is prepared by a process comprising the following sub-steps:
s21: dispersing the phosphorus silicon calcium bioactive glass in an aqueous solution of the hyaluronic acid and/or salt thereof to form a suspension,
S22: drying the suspension to a water content of about 0.05wt.% or less, and optionally subjecting the solid obtained after drying to grinding, thereby obtaining a composite particulate material comprising the phosphosilicate-calcium bioactive glass and the hyaluronic acid and/or salt thereof.
Scheme 8, the method for preparing a chronic healing deep skin wound protection repair composition according to scheme 7 above, wherein the concentration of the aqueous solution of hyaluronic acid and/or salt thereof in substep S21 is in the range of about 10 to about 15 wt%.
Scheme 9, the method for preparing a chronic healing deep skin wound protective repair composition according to scheme 7 or 8 above, wherein the drying in substep S22 comprises subjecting the suspension to freeze-drying, preferably comprising freezing the suspension at a temperature of about-18 to about-25 ℃ for about 4 to about 48 hours, and then drying it in a vacuum freeze dryer at a temperature of about-20 to about-50 ℃ at a vacuum of about 1.3 to about 13 pa.
Scheme 10, a chronic healing deep skin wound protective repair dressing formulation comprising a chronic healing deep skin wound protective repair composition according to any one of the above schemes 1 to 5 or a chronic healing deep skin wound protective repair composition prepared according to the method of any one of the above schemes 6 to 9.
Technical effects
Compared with the prior art, the technical scheme of the invention has the following unexpected technical effects:
(1) According to the chronic healing deep skin wound surface protection and repair composition, vaseline and/or lanoline are used as excipients and/or sustained release agents, hyaluronic acid and/or salt thereof with specific molecular weight are used as synergistic components of phosphorus-silicon-calcium bioactive glass, release of the hyaluronic acid and/or salt thereof and release of calcium (Ca), silicon (Si) and phosphorus (P) elements are effectively and stably controlled through proportion regulation and control of the components, and the composition can be safely and effectively used for chronic healing deep skin wound surface protection and repair in the mineralization process of the phosphorus-silicon-calcium bioactive glass. The composition for protecting and repairing the chronic healing deep skin wound surface is verified by in vitro experiments and animal experiments that the composition is reasonable in proportion, the phosphorus silicon calcium bioactive glass and the hyaluronic acid and/or the salt thereof are synergistic, the chronic healing wound surface can be promoted to heal, no stimulation is caused to skin tissues, and the effect is obvious compared with a single-component product.
(2) The chronic healing deep skin wound surface protection and repair composition can effectively control the amount of released elements after the phosphorus silicon calcium bioactive glass contacts with body fluid, and can keep the concentration of dissolved ions within a certain range through controlling the adding amount of the phosphorus silicon calcium bioactive glass, so that the stimulation reaction to wound tissues caused by over-fast and excessive ion dissolution is avoided. The phosphorus silicon calcium bioactive glass in the composition disclosed by the invention can generate hydration after being contacted with water, ions can be released in the hydration process, gel layers can be formed on the surface of the phosphorus silicon calcium bioactive glass, and then free ions are adsorbed to form hydroxyapatite. The study shows that the concentration of calcium ions is in an ascending trend within 24 hours, and the concentration of each element is basically balanced after 48 hours. The traditional phosphorus silicon sodium calcium bioactive glass powder is clinically contacted with body fluid, and then the local pH value is abnormally high, and the problem of irritation and inflammation is caused by excessive element release.
(3) The chronic healing deep skin wound protection and repair composition further expands the clinical application types and product forms of bioactive glass materials. The phosphorus silicon calcium bioactive glass adopted in the composition does not contain Na2 O, contains high-content SiO2 and CaO, ensures the activity and ion release of the bioactive glass through the control of specific surface area, simultaneously adopts hyaluronic acid with specific molecular weight and/or salt thereof to compound or adopts a vacuum freeze drying process to compound the two, ensures that the composition can uniformly release and effectively and safely when contacting wound surfaces, accelerates the wound surface healing, and has obvious clinical effect compared with single component. The elements released by the phosphorus-silicon-calcium bioactive glass in the composition have the effects of promoting cell proliferation and collagen growth; the released hyaluronic acid and/or salts thereof are capable of binding to fibrin, facilitating infiltration and transfer of cells into the matrix. The phosphorus-silicon-calcium bioactive glass with specific surface area and the hyaluronic acid with specific molecular weight and/or the hyaluronic acid salt with specific molecular weight are selected, so that the composition can be effectively released when contacting a wound surface, and simultaneously, the composition and the hyaluronic acid are compounded together by adopting a vacuum freeze-drying process, so that the composition can be uniformly released, and the effect of synergistically promoting wound surface healing is achieved.
(4) The phosphorus silicon calcium bioactive glass adopted in the chronic healing deep skin wound surface protection and repair composition is preferably porous granular material. The phosphorus silicon calcium bioactive glass is basically free of alkali metal oxide (such as sodium oxide), so that the phosphorus silicon calcium bioactive glass has relatively high content of calcium oxide, so that when a wound surface is coated, the instantaneous pH rise value of body fluid is reduced when the body fluid is contacted with the chronic healing deep skin wound surface protection and repair composition (the pH value of a soaking solution is less than 9 after the body fluid is soaked in water for 24 hours in the soaking proportion of about 0.1 g/mL), the irritation is reduced, the components in the composition play a synergistic effect, and the phosphorus silicon calcium bioactive glass can release Ca, si, P and other elements with biological effects when contacted with the body fluid, so that the growth factor expression and the cell proliferation are promoted, and the local microenvironment of a damaged part is improved.
(5) The chronic healing deep skin wound surface protection and repair composition can stay at the wound surface for at least 72 hours at the skin surface temperature, and through controlling the cone penetration of an excipient and/or a slow release agent, no liquid diluents such as glycerol, paraffin and the like are required to be additionally added into the composition, so that the problem that liquid components such as glycerol or paraffin and the like are separated out from a paste firstly due to unsuitable temperature is avoided, and meanwhile, the problem that the paste is harder and difficult to extrude and smear is also avoided.
(6) The preparation process of the chronic healing deep skin wound protection and repair composition is simple, and the obtained dressing medicament has long storage time. The composition is safe and convenient to use, has small dosage, can be smeared and used in a large area, can effectively protect and repair wound surfaces, and has better social and economic benefits.
(7) The chronic healing deep skin wound protection and repair composition disclosed by the invention does not contain polyethylene glycol commonly used in the prior art, does not cause local osmotic pressure to be too high or irritate, and is more beneficial to early cell proliferation. In addition, the composition does not contain carboxymethyl chitosan, carboxymethyl chitosan and/or polyethylene glycol which are commonly used in the prior art, and all have certain film forming property, and can form gel when meeting body fluid, so that sodium hyaluronate can be prevented from entering extracellular matrix to be effectively combined with protein.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described in the following in conjunction with the embodiments of the present invention. It should be apparent to those skilled in the art that the examples are provided only to aid in understanding the present invention and should not be construed as limiting the invention in any way. All other embodiments, which can be made by a person skilled in the art based on embodiments of the invention without any inventive effort, fall within the scope of protection of the invention. The process parameters for the specific conditions not noted in the examples below are generally as usual.
The endpoints of the ranges and any values disclosed in the present invention are not limited to the precise range or value, and the range or value should be understood to encompass a range or value close to the precise range or value. The term "about" as used herein means that the number to which it modifies may fluctuate within + -20%, + -15%, + -10%, + -5% or + -2% of the number. For numerical ranges, one or more new numerical ranges may be obtained in combination with each other between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point values, and are to be considered as specifically disclosed in the present invention.
According to a first aspect of the present invention, there is provided a chronic healing deep skin wound protective repair composition. The composition comprises or consists of phosphorus silicon calcium bioactive glass, hyaluronic acid and/or salts thereof, excipient and/or sustained release agent.
In the composition of the first aspect of the invention described above, the phosphosilicate bioactive glass contains substantially no alkali metal oxide, e.g., substantially no Na2 O. In the context of the present invention, the term "substantially free" or "free" of alkali metal oxide (or specifically Na2 O) is not to be understood as being "completely free" of alkali metal oxide (or specifically Na2 O) and is to be understood as alkali metal oxide (or Na2 O) which may contain unavoidable impurity levels or below a detection limit in the composition of the present invention. For example, in some embodiments of the invention, less than about 0.1 wt.%, preferably less than 0.05 wt.%, more preferably less than 0.01 wt.% of an alkali metal oxide, such as Na2 O, may be present in the chronic healing deep skin wound protective repair composition of the first aspect of the invention.
In the compositions of the first aspect of the invention described above, the phosphorus calcium silicate bioactive glass is present in an amount of about 4.5 to about 30.0 wt% (e.g., about 5 wt%, 7.5 wt%, 10 wt%, 12.5 wt%, 15.0 wt%, 17.5 wt%, 20.0 wt%, 22.5 wt%, 25 wt%, or 27.5 wt%) based on the total weight of the composition. Here, the content of the phosphosilicate-calcium bioactive glass should not be too low, e.g., not less than about 4.5 wt%, otherwise the composition releases too little phosphorus, silicon, and calcium elements after contact with body fluids, failing to achieve wound repair effects that promote growth factor expression and cell proliferation; the content of phosphorus silicon calcium bioactive glass should not be too high, for example, not more than about 30 wt%, otherwise the released phosphorus, silicon and calcium elements may be too high, causing irritation and inflammation problems of the wound surface.
In a preferred embodiment, the phosphosilicate-calcium bioactive glass has the form of a particulate porous material having a specific surface area of no less than about 200 m2/g (e.g., no less than about 220 m2/g、250 m2/g、270 m2/g、290 m2/g、310 m2/g、330 m2/g or 350 m2/g) and having a particle size in the range of about 10 to about 2000 μm (e.g., about 50 μm, 100 μm, 180 μm, 300 μm, 500 μm, or 750 μm). The particulate porous material should not have a surface area that is too low, such as not less than about 200 m2/g, which may otherwise result in inadequate contact of the surface of the phosphosilicate-calcium bioactive glass with bodily fluids, such that the phosphosilicate-calcium bioactive glass may not effectively mineralize the released elements in the vehicle or the mineralization rate may be slow. On the other hand, the particle size of the particulate porous material should not be too low, for example not less than about 10 μm, which might otherwise affect the uniformity of dispersion of the particulate porous material in the composition on the one hand, and on the other hand, might also lead to infiltration of inorganic powder particles into the tissue to be phagocytized by cells as foreign matter, resulting in rejection of the body, affecting cell activity; the particle size of the particulate porous material is also not too high, which may otherwise affect the spreadability of the composition paste. For example, when the particle size is greater than about 2000 μm, the composition paste may not only have a noticeable gritty feel when applied, but may also affect the appearance of the dressing paste.
The phosphorus silicon calcium bioactive glass preferably comprises about 46 wt.% to about 53 wt.% (e.g., about 47 wt.%, 48 wt.%, 49 wt.%, 50 wt.%, 51 wt.%, or 52 wt.%) SiO2.SiO2, based on the total weight of the phosphorus silicon calcium bioactive glass, is the most basic component in the glass, and glasses having SiO2 content within the defined range have good bioactivity and are capable of bonding well to soft skin tissue. Here, the SiO2 content should not be too low, for example not less than about 46% by weight, which might otherwise lead to a difficult formation of a glass network tetrahedral structure. The SiO2 content should also not be too high, for example not more than about 53% by weight, which might otherwise affect the activity of the glass. For example, it is well known in the art that when the SiO2 content exceeds about 70% by weight, it becomes an inert glass (such as building glass commonly found in life) and cannot be referred to as bioactive glass.
The phosphosilicate-calcium bioactive glass preferably comprises about 26 wt.% to about 43 wt.% (e.g., about 28 wt.%, 30 wt.%, 33 wt.%, 35 wt.%, 38 wt.%, or 40 wt.%) CaO and about 5 wt.% to about 28 wt.% (e.g., about 7 wt.%, 10 wt.%, 13 wt.%, 16 wt.%, 19 wt.%, 22 wt.%, or 25 wt.%) P2O5, based on the total weight of the phosphosilicate-calcium bioactive glass.
As well as P2O5 are the basic raw materials for forming the bioactive glass, their contents with the SiO2 component are mutually restricted in the phosphosilicate calcium bioactive glass of the present invention, and when CaO or P2O5 is too high or too low, it may affect the mineralization process of the bioactive glass and affect the ion release of the composition of the present invention after contacting body fluids.
In the composition of the first aspect of the present invention described above, the hyaluronic acid and/or salt thereof may comprise one or more of hyaluronic acid, sodium hyaluronate and zinc hyaluronate, and particularly preferably sodium hyaluronate.
In the composition of the first aspect of the invention described above, the hyaluronic acid and/or salt thereof is present in an amount of about 2 to about 10 wt% (e.g. about 4wt%, 6wt% or 8 wt%) based on the total weight of the composition. Here, the content of hyaluronic acid and/or salts thereof should not be too low, e.g. should not be less than about 2 wt%, otherwise dissolution of hyaluronic acid and/or salts thereof in the composition is difficult, and does not enter into the tissue for binding with fibrin; the content of hyaluronic acid and/or salts thereof should not be too high, for example not higher than about 10% by weight, otherwise the amount of hyaluronic acid and/or salts thereof eluted and the effect of binding to fibrin has not been significantly changed on the one hand; on the other hand, the appearance and stability of the composition may be affected, and the viscosity of the composition may be deteriorated.
The hyaluronic acid and/or salt thereof preferably has the form of a particulate material, the particulate material preferably having a particle size of not more than about 220 μm, preferably in the range of about 70 to about 180 μm (e.g. about 80 μm, 100 μm, 120 μm, 140 μm or 160 μm). The particle size of the hyaluronic acid and/or salt thereof is also not too high, e.g., not greater than about 220 μm, which would otherwise affect the spreadability of the composition, would have a noticeable gritty feel when the composition is spread, and may also affect the appearance of the dressing paste formed by the composition, and the composition stability would be poor; the particle size of the hyaluronic acid and/or salt thereof is preferably not too low, e.g. not less than about 70 μm, which might otherwise affect the uniformity of dispersion and dissolution rate of the hyaluronic acid and/or salt thereof.
The hyaluronic acid and/or salt thereof preferably has a molecular weight in the range of about 200 to about 1200 KDa (e.g., about 400, 600, 800, 1000, KDa), which is not too low, e.g., not less than about 200, KDa, otherwise the hyaluronic acid and/or salt thereof is susceptible to degradation; the molecular weight of the hyaluronic acid and/or salt thereof is also preferably not too high, e.g. not higher than about 1200 KDa, otherwise the hyaluronic acid and/or salt thereof is not easily penetrated into skin tissue, affecting its binding to fibrin.
In another preferred embodiment, the phosphosilicate-calcium bioactive glass and the hyaluronic acid and/or salt thereof together form a composite particulate material. The composite particulate material may be prepared as follows: the particulate porous phosphosilicate-calcium bioactive glass is first dispersed in an aqueous solution of the hyaluronic acid and/or its salt to form a suspension, then the suspension is dried to a water content of about 0.05 wt.% or less, and the solid obtained after drying is optionally subjected to grinding, thereby obtaining a composite particulate material comprising the phosphosilicate-calcium bioactive glass and the hyaluronic acid and/or its salt. Here, such a process results in the obtained composite particulate material having the particle size and porosity of the particulate porous phosphosilicate bioactive glass used, which has a layer of hyaluronic acid and/or salts thereof on the surface thereof, so that upon encountering body fluids, the hyaluronic acid and/or salts thereof dissolve out first without affecting the activity of the phosphosilicate bioactive glass. One major advantage of the composite particulate material is that it ensures that the active substances released by the composition are at the same base point, avoiding the possible maldistribution of hyaluronic acid and/or its salts when they are dispersed separately from the phosphorus-silicon-calcium bioactive glass.
In the above-described composition of the first aspect of the present invention, an excipient and/or a sustained-release agent is present. The excipient and/or sustained release agent may comprise one or more substances that act to shape the composition and/or to sustained release the components of the composition in the composition. The excipient and/or sustained-release agent may be an excipient and/or sustained-release agent commonly used in the pharmaceutical field. In view of the ranges of content of phosphosilicate bioactive glass and hyaluronic acid and/or salts thereof that are desirably included in the compositions of the present invention as described above, the amount of the excipient and/or slow-release agent in the composition is in the range of about 60 to about 92.5 wt.% (e.g., about 67 wt.%, 70 wt.%, 75 wt.%, 80 wt.%, or about 86 wt.%) based on the total weight of the composition.
Additionally, in a preferred embodiment, the excipient and/or sustained release agent preferably has an cone penetration in the range of about 160 to about 190, preferably in the range of about 170 to about 185 (e.g., about 175 or 180). In the context of the present invention, the cone penetration of the excipient and/or slow release agent is determined according to GB/T269-91. From the aspect of clinical use of the composition paste, the penetration degree of the excipient and/or the slow release agent is not too small, for example, is not less than 160, otherwise, on one hand, the hardness of the composition paste is increased, so that the composition paste is difficult to push out of a plastic-aluminum hose or a syringe on the market, and on the other hand, the application force of the composition paste is increased, and the pain of a wound surface is increased; the penetration of the excipient and/or slow release agent should not be too great, e.g. not exceeding 190, otherwise the composition paste would become too soft and too thin, and the residence time at the wound coverage would be shortened, resulting in frequent dressing change cycles.
In a preferred embodiment, the excipients and/or sustained release agents included in the compositions of the present invention preferably comprise one or more of petrolatum and lanolin. Vaseline and lanolin are used as oily matrixes of the composition, so that an oil film can be formed at a wound surface, a certain moisturizing effect is achieved, and invasion of external microorganisms to the wound can be prevented to a certain extent. In addition, petrolatum and lanolin can generate a synergistic effect with the phosphosilicate calcium bioactive glass and hyaluronic acid and/or its salt contained in the chronic healing deep skin wound protection and repair composition of the present invention, so as to effectively control the release of calcium (Ca), silicon (Si) and phosphorus (P) elements and the release of hyaluronic acid and/or its salt in the phosphosilicate calcium bioactive glass. In a particularly preferred embodiment, the excipient and/or slow release agent preferably comprises lanolin and petrolatum in a weight ratio in the range of about (0-18): 100, preferably about (2-10): 100. Lanolin and vaseline in the above ratio range can achieve the technical effect of the present invention, in particular.
According to a second aspect of the present invention there is provided a method for preparing a chronic healing deep skin wound protective repair composition according to the first aspect of the present invention as described above.
In a method of the second aspect of the invention, the method comprises the steps of:
s1: liquefying the excipient and/or the sustained release agent, and
S2: dispersing the phosphorus silicon calcium bioactive glass and the hyaluronic acid and/or the salt thereof in the liquefied excipient and/or the sustained release agent obtained in the step S1 sequentially or simultaneously according to any sequence, thereby obtaining the chronic healing deep skin wound protection and repair composition.
The preferred features described above in relation to the specific compositional features in the first aspect of the invention are also applicable to the method of the second aspect of the invention.
In addition, in the case where the phosphosilicate-calcium bioactive glass and the hyaluronic acid and/or salt thereof together form a composite particulate material in the composition of the above first aspect, the method of the second aspect of the invention comprises the steps of:
S1: liquefying the excipient and/or the sustained release agent,
S2: dispersing the composite particulate material in the liquefied excipient and/or slow release agent obtained from step S1, thereby obtaining the chronic healing deep skin wound protective repair composition, wherein the composite particulate material is prepared by a process comprising the following sub-steps:
s21: dispersing the phosphorus silicon calcium bioactive glass in an aqueous solution of the hyaluronic acid and/or salt thereof to form a suspension,
S22: drying the suspension to a water content of about 0.05wt.% or less, and optionally subjecting the solid obtained after drying to grinding, thereby obtaining a composite particulate material comprising the phosphosilicate-calcium bioactive glass and the hyaluronic acid and/or salt thereof.
In the substep S21 of the above method, the concentration of the aqueous solution of hyaluronic acid and/or a salt thereof is not particularly limited, but in a preferred embodiment, the concentration is in the range of about 10 to about 15% by weight.
In step S22 of the above method, the drying operation is not particularly limited, but in a preferred embodiment, the drying comprises subjecting the suspension to freeze-drying, preferably comprising freezing the suspension at a temperature of about-18 to about-25 ℃ for about 4 to about 48 hours, and then drying it in a vacuum freeze dryer at a temperature of about-20 to about-50 ℃ at a vacuum of about 1.3 to about 13 pa.
According to a third aspect of the present invention there is provided a chronic healing deep skin wound protective repair dressing formulation comprising a chronic healing deep skin wound protective repair composition according to the first aspect of the present invention as described above or prepared according to the method of the second aspect of the present invention as described above.
The chronic healing deep skin wound protective repair dressing formulation of the third aspect of the invention may be in a form common in the art of medical dressing formulations including, but not limited to, ointments, plasters, gels, cataplasms, patches and the like. The preparation of the chronic healing deep skin wound surface protective and repair dressing can comprise any common auxiliary materials of the preparation besides the chronic healing deep skin wound surface protective and repair composition, including but not limited to a back lining, release paper, a packaging bag, a hose, a syringe and the like.
In addition, the preferred features described above in relation to the composition specific technical features in the first and second aspects of the invention are also applicable to the formulation of the third aspect of the invention.
Examples and comparative examples
The technical scheme of the present invention will be described in further detail with reference to specific examples and comparative examples.
The starting materials used in the examples and comparative examples:
Phosphorus silicon calcium bioactive glass: commercially available from beijing happy probiotic new material limited, prepared by a sol-gel process. And (3) sieving the phosphorus silicon calcium bioactive glass with an 80-mesh sieve, collecting powder below the sieve, sieving the collected powder with a 100-mesh sieve, and collecting powder on the sieve, wherein the powder is the phosphorus silicon calcium bioactive glass raw material used in the embodiment of the invention. The phosphorus silicon calcium bioactive glass raw material is in the form of a porous granular material, has a particle size ranging from about 150 mu m to about 180 mu m, has a specific surface area of about 309 m2/g according to GB/T19587-2004, and comprises the following components based on the total weight of the phosphorus silicon calcium bioactive glass:
About 51 wt% SiO2;
about 33 weight percent CaO; and
About 16 wt% P2O5.
Phosphorus silicon sodium calcium bioactive glass: commercially available from beijing happy probiotic new material limited, prepared by a sol-gel process. And (3) sieving the phosphorus silicon calcium bioactive glass with an 80-mesh sieve, collecting powder below the sieve, sieving the collected powder with a 100-mesh sieve, and collecting powder on the sieve, wherein the powder is the phosphorus silicon calcium bioactive glass raw material used in the embodiment of the invention. The phosphorus silicon calcium bioactive glass raw material is in the form of a porous granular material, the grain diameter is in the range of about 150 mu m to about 180 mu m, the specific surface area is about 309 m2/g according to GB/T19587-2004 test, and the composition comprises the following components based on the total weight of the phosphorus silicon sodium calcium bioactive glass:
About 45 wt% SiO2;
about 24.5 weight percent CaO;
About 6wt% P2O5; and
About 24.5 wt% Na2 O.
Sodium hyaluronate: commercially available from Huaxi biotechnology limited and having a molecular weight of about 520 KDa. The sodium hyaluronate is sieved by a 80-mesh sieve, then the powder below the sieve is collected, the collected powder passes through a 100-mesh sieve, and the powder on the sieve is collected, wherein the powder is the sodium hyaluronate powder material used in the embodiment and the comparative example of the invention. The sodium hyaluronate powder material has a particle size in the range of about 150 μm to about 180 μm.
Vaseline: commercially available from Nanchang white cloud pharmaceutical Co., ltd, and has a cone penetration of about 171 as measured according to GB/T269-91.
Lanolin: commercially available from Shanghai Huating lanolin works Co., ltd, has a cone penetration of about 179 as measured according to GB/T269-91.
The invention relates to a preparation of a chronic healing deep skin wound surface protection and repair composition
Example 1:
the formula comprises the following components: about 10 parts by weight of phosphorus silicon calcium bioactive glass, about 4 parts by weight of sodium hyaluronate, about 78 parts by weight of vaseline, about 8 parts by weight of lanolin, and about 40 parts by weight of purified water were prepared.
The preparation process comprises the following steps: the about 4 parts by weight of sodium hyaluronate was dissolved in the about 40 parts by weight of purified water to form an aqueous sodium hyaluronate solution. The approximately 10 parts by weight of the phosphorus silicon calcium bioactive glass porous particulate material was then uniformly dispersed in the aqueous solution of sodium hyaluronate. The resulting suspension was frozen in a freezer at about-20 c for about 24 hours and then lyophilized to a water content of less than or equal to 0.05 wt% using a vacuum freeze dryer at a temperature of about-50 c and a vacuum of about 1.3 to 13 pa. Grinding the solid substance obtained after freeze-drying to finally obtain the composite freeze-dried powder of the phosphorus calcium silicate bioactive glass and the sodium hyaluronate with the particle size ranging from about 150 mu m to about 180 mu m. The mixture comprising the about 78 parts by weight of petrolatum and the about 8 parts by weight of lanolin is fully liquefied at a temperature of about 60 ℃. And then adding the composite freeze-dried powder into the liquefied mixture of the vaseline and the lanoline, fully stirring and homogenizing, cooling to room temperature, and filling and sealing to obtain the composition of the embodiment 1.
Example 2:
the formula comprises the following components: about 25 parts by weight of phosphorus silicon calcium bioactive glass, about 8 parts by weight of sodium hyaluronate, and about 67 parts by weight of vaseline were prepared.
The preparation process comprises the following steps: the about 67 parts by weight of petrolatum was completely liquefied at a temperature of about 60 ℃. And then adding the phosphorus silicon calcium bioactive glass porous granular material with the weight of about 25 parts into the liquefied vaseline, fully stirring and homogenizing, then adding the sodium hyaluronate powder material with the weight of about 8 parts, fully stirring and homogenizing, cooling to room temperature, and filling and sealing to obtain the composition of the embodiment 2.
Preparation of skin wound surface protecting and repairing composition of comparative example
Comparative example 1 and comparative example 2:
The formulation and formulation process were similar to examples 1 and 2, respectively, described above, except that the phosphorus silicon calcium bioactive glass porous particulate material in examples 1 and 2 was replaced with a phosphorus silicon sodium calcium bioactive glass material.
In vitro experiments
In order to verify the physical and chemical performance parameters of the compositions of examples 1 and 2 and comparative examples 1 and 2 of the present invention, the compositions of examples 1 and 2 and comparative examples 1 and 2 of the present invention were subjected to in vitro experiments.
(1) PH determination
About 1.5g of the compositions from inventive examples 1 and 2 and comparative examples 1 and 2 were hermetically soaked in a suitable container with about 15: 15 mL purified water at a temperature of about 37.+ -. 1 ℃ for about 24 hours. Taking out the aqueous liquid after the sample is soaked, and uniformly mixing to obtain the test liquid. And measuring the pH value of the test liquid by an acidometer. The measured pH results were:
Example 1: about 7.6; example 2: about 8.3;
comparative example 1: about 9.1; comparative example 2: about 9.7.
As can be seen from the above pH measurements, the compositions of examples 1 and 2 of the present invention have a pH of less than about 9.0, even less than about 8.5, e.g., up to only about 8.3, after being fully soaked in purified water at a temperature of about 37±1 ℃ (soaking ratio of about 0.1 g/mL), which is more similar to the pH of human body fluids, greatly reducing the possibility of severe irritation to the skin wound surface of the human body. The compositions of comparative examples 1 and 2, after having been fully soaked in purified water (soaking ratio of about 0.1 g/mL), had a pH of above about 9.0, even up to about 9.7.
(2) Determination of the concentration of dissolved sodium hyaluronate
The sodium hyaluronate content in the test solution obtained in (1) was tested by inductively coupled plasma atomic emission spectrometry (ICP-AES), and the test result was:
Example 1 sodium hyaluronate concentration was about 0.16 g/mL;
example 2 sodium hyaluronate concentration was about 0.28 g/mL;
Comparative example 1 sodium hyaluronate concentration was about 0.16 g/mL;
Comparative example 2 sodium hyaluronate concentration was about 0.28 g/mL.
From the above measurement results of sodium hyaluronate concentration, it is understood that the compositions of examples 1 and 2 and comparative examples 1 and 2 of the present invention can release sodium hyaluronate after being sufficiently soaked in purified water (soaking ratio of about 0.1 g/mL), and the release effect is substantially the same without being affected by bioactive glass. It can thus be deduced that sodium hyaluronate in the compositions of examples and comparative examples may pass smoothly through body fluids into the cytoplasmic matrix after contact with body fluids.
(3) Determination of the concentration of released elements
Leaching experiments were performed on samples of the compositions of examples 1 and 2 and comparative examples 1 and 2 of the present invention, using purified water. During the leaching process, the leaching ratio of the composition to be leached with purified water was set to about 0.2 g/mL. Test sample solutions were withdrawn at 24 hours, 48 hours and 72 hours of extraction sample time points, respectively. The concentration of the elements in the leaching solution was tested by inductively coupled plasma atomic emission spectrometry (ICP-AES). The test results are shown in table 1 below:
From the above-described experimental results of the element concentration release measurement, the compositions of examples 1 and 2 according to the present invention can effectively and smoothly control the release of calcium (Ca), silicon (Si) and phosphorus (P) elements during mineralization of the phosphorus-silicon-calcium bioactive glass, and can maintain the concentration of dissolved ions within a certain range by controlling the addition amount of the phosphorus-silicon-calcium bioactive glass. For example, the composition of the present invention can achieve a fluctuation of the release amount of Ca, si and P elements within 30% in 24 to 72 hours at a leaching ratio of about 0.2 g/mL, for example, specifically, ca element fluctuation coefficients in examples 1 and 2 are only about 1.17 and 1.125, respectively (calculation of fluctuation coefficient is 72 hours ion concentration divided by 24 hours ion concentration). The composition of the embodiment of the invention avoids the stimulation reaction caused by the excessive dissolution of ions to the wound tissues, thereby achieving the expected technical effect in the animal experiment process as described below.
In the body fluid of wound tissue, ca, si and P elements with a certain concentration have biological effect. For example, elemental Si can promote gene repair, anti-inflammatory and antibacterial, aid blood circulation and rejuvenate wounds; stimulating the formation of type I collagen, modulating type I and type III collagen; elemental Ca ions are a very important intracellular messenger and play an important role in signaling, regulating various functions of cells such as proliferation, differentiation, etc. However, when the element concentration exceeds a certain limit, the effect is reduced and even cell growth is affected. As for the specific critical value of the element concentration, no specific research data is given in the prior art, and the literature reports that Ca2+ concentration exceeding 300 mg/L can negatively affect cell proliferation; a silicon concentration exceeding 1.67 mmol/L has an inhibitory effect on cell proliferation.
The compositions of comparative example 1 and comparative example 2, at a leaching ratio of about 0.2 g/mL, have a large fluctuation of Ca element within 24 to 72 hours in the composition, and a fluctuation coefficient of much more than 1.5, because the bioactive glass containing the alkali metal sodium oxide preferentially releases a large amount of Na ions when mineralized in contact with body fluid, and the release amounts of calcium, silicon and phosphorus element are relatively low. In addition, since the phosphorus silicon sodium calcium bioactive glass in the comparative example composition contains a large amount of sodium oxide components, release of a large amount of sodium ions after contact with body fluid can cause excessive Na element content in the local part of the wound surface. The proper amount of Na element can generally maintain the osmotic pressure of the cell fluid, and the excessive content of Na element can cause the osmotic pressure imbalance of the cell fluid, which is unfavorable for the growth of wound cells.
(4) Shape forming test
The compositions from inventive examples 1 and 2 and comparative examples 1 and 2 were uniformly coated on glass plates with a coating area of about 5 cm ×5 cm =25 cm2, giving a test sample of about 0.15 g/cm2. The test samples were placed vertically in a constant temperature and humidity cabinet at about 37 ℃ and a relative humidity of about 64% for about 72 hours to simulate the conditions of the compositions comprising examples 1 and 2 of the present invention and comparative examples 1 and 2 at vertical wound surfaces. The samples to be tested comprising the compositions of examples 1 and 2 according to the invention and comparative examples 1 and 2 were then observed for the flow phenomenon.
From the experimental results observed, it is clear that the samples to be tested comprising the compositions of examples 1 and 2 of the present invention and comparative examples 1 and 2 are able to stay in situ at the skin surface temperature (about 37 ℃) for at least about 72 hours under the conditions of the simulated wound surface.
Animal experiment
To compare the effect and effect of the phosphorus silicon calcium bioactive glass powder, the sodium hyaluronate gel and the compositions of examples 1 and 2 and comparative examples 1 and 2 of the present invention, animal experiments were conducted to verify.
Healthy Bama pigs 7 were selected, randomly grouped, with each Bama pig corresponding to one group of following drug samples.
The animal experiments were grouped as follows:
Group ①: sterile gauze (control group)
Group ②: 4% sodium hyaluronate hydrogel+sterile gauze
Group ③: phosphorus silicon calcium bioactive glass powder and sterile gauze
Group ④: composition of example 1 of the invention + sterile gauze
Group ⑤: composition of inventive example 2 + sterile gauze
Group ⑥: composition of comparative example 1 + sterile gauze
Group ⑦: composition of comparative example 2 + sterile gauze
And (3) preparing the back skin after general anesthesia of each balm Ma Zhu, and cutting square full-layer skin defect wound surfaces along two sides of a median line of the back under a strict aseptic operation technology after local disinfection, wherein the length and width of each wound surface are about 5cm, the depth is about 2 cm, 3 wound surfaces are arranged on each side, the interval between the wound surfaces is about 8 cm, and 6 chronic deep skin wound surfaces are formed on each balm pig. The model in the animal experiment is a full-layer skin defect model, namely, defects occur in the three-layer structure of skin, namely, the surface layer, the dermis layer and the subcutaneous tissue, and the wound surface of the defects has long healing time period, so the model can be defined as a chronic healing (or difficult healing) deep skin wound surface. After the wound surface is established, the wound surface is subjected to hemostasis and disinfection, and after hemostasis, the skin wound surface of the Bama pig is coated with the five groups of medicament products, wherein the coating amount is about 0.15 g/cm2. The application area of the medicament is slightly larger than the wound surface area, and then the wound surface is wrapped and fixed by using sterile gauze. The medicine was changed every 7 days, and the wound repair was recorded (with scale, photographed at the same height and angle). And finally, performing healing rate calculation.
The results of the observation of the wound surface of the animal test are as follows:
group ①: in the first week after operation, exudation is obvious, and a small amount of bleeding exists; in the second week after operation, there is still obvious exudation, scabbing begins to appear, and bleeding occurs during dressing change; in the fifth week after surgery, 2 wounds showed infection, and no wound showed complete healing.
Group ②: in the first week after operation, exudation is obvious, and a small amount of bleeding exists; in the second week after operation, there is still obvious exudation; in the fifth week after surgery, mild infection occurred in 2 wounds, and 1 wound healed completely.
Group ③: a small amount of exudation occurs in the first week after operation; at week 2 post-operation, 1 wound had inflammatory material and surrounding skin was slightly red; in the fifth week after the operation, 2 wounds were substantially completely healed. In the dressing change process, a small amount of powder remains on the wound surface.
Group ④: in the first week after operation, a small amount of exudation exists, the wound is free from infection, fresh granulation tissue grows faster, and wound surface shrinkage and healing are faster; in the fifth week after the operation, 6 wounds were substantially completely healed.
Group ⑤: in the first week after operation, a small amount of exudation exists, the wound is free from infection, fresh granulation tissue grows faster, and wound surface shrinkage and healing are faster; five weeks after the operation, 5 wounds were substantially completely healed.
Group ⑥: in the first week after operation, a small amount of exudation exists, no infection phenomenon occurs in wounds, in the second week after operation, obvious exudation still exists, inflammatory substances appear in 1 wound in the third week after operation, the wound surface is obviously reduced in the fifth week after operation, and 3 wound surfaces are basically and completely healed.
Group ⑦: a small amount of exudation exists in the first week after operation, no infection phenomenon occurs in wounds, obvious exudation still exists in the second week after operation, yellow inflammatory substances appear on 2 wounds, the surrounding skin is slightly red and swelling, and slight bleeding appears when dressing change is performed; in the fifth week after operation, the wound surface is obviously reduced, and 1 wound surface is basically and completely healed.
And (3) calculating the wound healing rate:
In the animal experiment process, the wound surface is recorded by photographing (photographing with a scale at the same height and angle) on the 0 th day, the 7 th day, the 14 th day, the 21 th day, the 28 th day and the 35 th day after the wound surface is formed, and the wound surface area of each stage is calculated. The calculation results of the average wound healing rate are shown in figure 1 and table 2 below. The calculation formula of the wound healing rate is as follows:
R = (A0-At)/At×100%
Wherein the method comprises the steps of
R is the wound healing rate;
A0 is the area of the wound surface at the initial time;
At is the area of the wound At time t.
The average wound healing rate calculation results are shown in table 2:
as shown in fig. 1 and table 2, the rate of skin wound healing of the pamphlet increased over time for each group of agents, over treatment with seven groups of agent samples. However, the control group had the lowest wound healing rate, and the phosphorus-silicon-calcium bioactive glass powder of group ③ and the sodium hyaluronate of group ② had a certain promoting effect on the wound healing stage, but had the disadvantage of having a longer healing time for chronic deep wounds, which may cause problems of irritation and infection. The compositions of groups ④ and ⑤ of examples 1 and 2 of the present invention promote healing, have a significant therapeutic effect on promoting healing of large-area deep skin wounds, and the rate of healing is the fastest, without any infection being found to occur. Of these, group ④ is better than group ⑤, probably due in part to the fact that the phosphosilicate-calcium bioactive glass and the sodium hyaluronate together form a composite particulate material. The compositions of groups ⑥ and ⑦ of comparative examples 1 and 2 also promote healing, but also have some irritation to large-area deep skin wounds, resulting in less wound healing than the compositions of the examples of the present invention, and no desired effect is achieved.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and not for limiting the same; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some or all of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the essence of the corresponding technical solutions.