CN109985271B - Composite collagen dressing for healing-difficult wound repair and preparation method thereof - Google Patents
Composite collagen dressing for healing-difficult wound repair and preparation method thereof Download PDFInfo
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
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- A—HUMAN NECESSITIES
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- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
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Abstract
The invention discloses a composite collagen dressing for healing-difficult wound repair and a preparation method thereof, which is prepared by dispersing type I collagen and type III collagen in a solvent according to a proportion, homogenizing, compounding and reshaping; the components related by the invention are selected from multi-component collagen materials with different types and different contents, so that the characteristics of each collagen component can be fully exerted, the repair of the wound is accelerated, and the wound healing quality is improved.
Description
Technical Field
The invention belongs to the field of biological materials, and particularly relates to a composite collagen dressing for healing-difficult wound repair and a preparation method thereof.
Background
The prevalence of various chronic diseases increases year by year, resulting in high morbidity on chronic wounds such as pressure sores, diabetic feet, venous leg ulcers and the like associated therewith. The clinical common wounds are mainly changed from acute wounds to chronic wounds, and the chronic wounds caused by various wounds, burns, scalds, non-healing wounds after operation, bedsores, foot rot and ecthyma (commonly called leg rot) jump the first place of morbidity.
Diabetes is a frequently-occurring disease and a common disease with high morbidity, great harm and wide spread worldwide, is one of civilized diseases of modern life, and is listed as one of three difficult and complicated diseases by WHO (world health organization). At present, 1.25 million diabetics exist in the world, the number of diabetics reaches 4000 million, and the diabetic foot is one of common chronic complications of diabetes and is also a main reason for amputation and disability of the diabetics. The diabetic foot is characterized in that the protective function of the lower limb of the foot of a diabetic patient is reduced due to neuropathy, and the foot of the diabetic patient is ulcerated and gangrene can be generated due to microcirculation disturbance caused by insufficient arterial perfusion caused by macrovascular and microvascular lesions, so that the diabetic foot not only causes pain to the patient, but also adds huge economic burden to the diabetic patient.
The treatment of chronic ulcer wounds of diabetic feet essentially initiates a cascade of "waterfall" reactions of wound healing by debridement, which converts chronic wounds to acute wound states. Different debridement treatments are carried out according to the gangrene wound surfaces of different types and different stages, so that an opportunity of correctly appearing healing processes according to time sequence is provided for cells, and the healing of the wound surfaces is promoted. However, the treatment means such as pharmaceutical intervention and surgical intervention still need self-healing capability, so that the improvement of self-healing capability of the wound surface is a necessary condition for treating diabetic foot.
Trauma is the destruction of tissue or organs by mechanical factors added to the human body. In the present day that the deep trauma rate is gradually increased and the trauma situation is gradually complicated and serious, the deep trauma is extremely extensive and is often accompanied by the damage of important organs, and more blood or plasma loss, severe anoxia, acidosis, massive tissue necrosis and the like exist. These intense stimuli contribute to tissue degeneration, thereby accelerating the rate of metabolic changes, leading to severe complications such as shock, acute renal failure, sepsis, and the like. In the face of such wounds, treatment and repair of the damaged area is required after the rescue has been performed.
In the healing process of the two wound tissues, infection and adhesion are often caused, and antibiotics, anti-adhesion membranes and hemostatic products are generally adopted to act on the wound to solve the problems. Most of the anti-adhesion and hemostatic products comprise components such as polylactic acid, chitosan, starch, gelatin and the like, and promote the healing of body tissues.
As a natural polymer material, the collagen has the advantages of high mechanical property, good biocompatibility, low immunogenicity, biodegradability and hemostatic effect, and is a biomedical material with high application value. At present, studies have found that the human body contains twenty or more types of collagen, each of which adds a specific function to the structural action of collagen. The different types of collagen play different roles:
type I collagen is the most abundant and studied collagen, and is mainly present in connective tissues such as skin, bones, blood vessels, and the like.
Type III collagen is widely distributed in tissues containing type III collagen other than bone, and is an important component in reticular fibers of interstitial tissues of lung, liver, dermis, spleen, and blood vessels.
For the repair of the wound surface which is difficult to heal, the collagen with single component can play a repair role, but the action time is long. If the two-component collagen dressing containing two different types of collagen is prepared by compounding the different types of collagen, and the characteristics of each collagen component are fully exerted, the wound repair is certainly promoted, the wound repair time is effectively reduced, and the two-component collagen dressing has an important application value for improving the treatment level of a wounded person or a patient.
Disclosure of Invention
Based on the above defects and requirements objectively existing in the field, the invention aims to prepare a composite collagen raw material for repairing a wound which is difficult to heal, makes up the blank of preparation and combined application of different types of composite collagens at present, and enhances the versatility of the collagen, so that the collagen has a wider application range. The raw materials of the composite glue with different formulas can be selected according to the position of the wound, so that the wound healing is promoted, and the healing quality is improved.
The technical scheme of the invention is as follows:
a composite collagen dressing for repairing a wound surface which is difficult to heal is characterized by being compounded by two different types of collagen;
the different types of collagen are selected from type I collagen and type III collagen.
The type I collagen is selected from bones, skins and bovine achilles tendons of human or animals; the type III collagen is selected from human or animal skin, blood vessel wall, and amnion.
The proportion of the type I collagen and the type III collagen is 99.99-85: 0.01-15.
The dosage form of the composite collagen dressing is selected from: sponge-like materials, fibrous membranes, layered membranes;
preferably, the refractory wound surface is: the self-healing capability is poor, and the healing time is more than 2 months of wound surface;
more preferably, the refractory wound is selected from the group consisting of: chronic or traumatic wound surface with difficulty in healing.
The preparation method of the composite collagen dressing for repairing the wound surface which is difficult to heal is characterized by comprising the following steps: dispersing type I collagen and type III collagen in a solvent in proportion, homogenizing, compounding, and molding.
Dispersing 99.99-85 parts of type I collagen and 0.01-15 parts of type III collagen in a solvent according to a proportion, homogenizing, compounding and then molding;
preferably, the solvent is distilled water, diluted acetic acid, trifluoroacetic acid, hexafluoroisopropanol; the homogenate refers to mixing homogenate by a high-speed mixer;
the dosage form of the composite collagen dressing is a fibrous membrane; the homogenization step of the fiber membrane refers to: preparing uniform slurry with certain mass concentration; the forming step includes: adding a cross-linking agent into homogenate with a certain mass concentration to prepare a collagen copolymer through cross-linking, dissolving the cross-linked copolymer into a solvent, placing the solution into a liquid storage tank of a high-voltage electrostatic spinning machine, and preparing a chemical composite collagen fiber membrane through electrostatic spinning;
the dosage form of the composite collagen dressing is a layered film; the homogenization step of the layered membrane refers to: preparing two homogenates with different collagen ratios; the forming step includes: evenly spreading I and III collagens with the weight ratio of 99.3:0.7 in a container, and pre-freezing the container in an ultra-low temperature refrigerator at the temperature of-80 ℃ for 24 hours to form an outer-layer even pulp membrane; then, evenly spreading the I and III collagen homogenates with the weight ratio of 97:3 on an outer homogenate membrane, pre-freezing the homogenate membrane at the temperature of minus 80 ℃ for 24 hours to form a contact layer homogenate membrane, and drying the contact layer homogenate membrane in a freezing vacuum dryer at the temperature of minus 50 ℃ for 24 hours to prepare a layered membrane;
the content of collagen III in the outer-layer homogenizing film is 0.01-1%, and the content of collagen III in the contact-layer homogenizing film is 1-15%.
The product of the invention has the following characteristics:
(1) the composite collagen dressing prepared by the invention is prepared by compounding different types of collagen according to a certain proportion, the proportion of the two types of collagen can be adjusted according to the severity of wound surfaces, and the composite collagen dressing can be suitable for different wound surfaces.
(2) The composite collagen dressing prepared by the invention can be prepared into different specifications and forms according to different forming processes, and can be applied to different parts and wound surfaces, so that the application range of the composite collagen dressing is further enlarged.
(3) The composite collagen dressing prepared by the invention has the advantages of easily available raw materials, simple and controllable preparation process and capability of realizing industrialization.
In conclusion, the invention aims at the application requirements of different parts and different degrees of wound surfaces; the components involved in the invention are all different types of collagen, and no foreign matter is introduced, so that the low immunogenicity and good biocompatibility of the collagen can be maintained; but also can reflect different functions of different types of collagen at the same time, and is a raw material of the multifunctional composite glue; and the raw material of the composite gel modified by chemical crosslinking can be selected according to different severity degrees of wounds, so that the degradation time of the collagen is prolonged, and the collagen can be more effectively used for wound surfaces.
Drawings
FIG. 1 is an electron micrograph, at 100 magnification, of collagen type I alone;
fig. 2 is an electron micrograph of a collagen type I + collagen type III composite dressing provided according to an embodiment of the present invention, at 100 times magnification.
FIG. 3 is a graph showing HE staining of rat dorsal fascia (deep II degree wound model repair)
Detailed Description
The present invention is further illustrated by the following examples, which are intended to be illustrative, not limiting and are not intended to limit the scope of the invention.
Sources of biological material
The skin, bovine achilles tendon, blood vessel wall, amniotic membrane of the animal can be obtained from a farm, a slaughterhouse, or are commercially available; the human skin, vascular wall, and amniotic membrane can be obtained from donor and hospital.
Rat Material of Experimental example 3 the composite collagen dressing for healing-resistant wound repair of the present invention, group 1 example, was provided by the Experimental animal center of Tianjin medical university
The present group of embodiments provides a composite collagen dressing for healing a difficult-to-heal wound, which herein refers to a wound with poor self-healing ability and healing time longer than 2 months, and generally includes chronic difficult-to-heal wound and traumatic difficult-to-heal wound.
All embodiments of this group share the following common features:
the weight ratio of the type I collagen to the type III collagen in the composite collagen dressing is 99.99-85: 0.01-15;
although the amount of the type III collagen can be as low as 0.01, the type III collagen is trace, but the function of the type III collagen in the composite collagen auxiliary material is quite relevant, and the inventor surprisingly finds that the addition of a small amount or trace amount of the type III collagen can effectively promote the regeneration of collagen, accelerate the formation of microvessels, promote the vascularization, and simultaneously facilitate the repair of wound surfaces and the removal of scars.
The type I collagen plays a role in the healing effect of the wound surface which is difficult to heal: the biological growth factor carrier has considerable biomechanical property, has strong affinity among different cells, has special affinity with growth factors playing a key role in the healing process of wounds, has the function of a signal transfer station, and can intercept, store and transport the growth factors, promote wound healing and tissue repair.
The type I collagen is selected from bones, skins and bovine achilles tendons of human or animals;
the type III collagen is selected from human or animal skin, blood vessel wall, and amnion.
The dosage form of the composite collagen dressing is selected from: sponge-like materials, fibrous membranes, layered membranes.
In a specific embodiment, the composite collagen dressing can be used for repairing diabetic feet and deep wounds and comprises the following collagen in parts by weight, and according to the type and severity of a wound surface, the content of type I collagen in the composite collagen dressing is gradually reduced, and the content of type III collagen in the composite collagen dressing is gradually increased.
Table 1 composite collagen dressing with different proportions
Group 2 example, preparation of composite collagen dressing for healing-refractory wound repair of the invention
The present group of embodiments provides a method of preparing a composite gum material as described in any of the embodiments of group 1. All embodiments of this group have the following common features: the preparation method comprises the following steps: dispersing type I collagen and type III collagen in a solvent in proportion, homogenizing, compounding, and molding.
In a specific embodiment, the preparation method comprises the following steps: 99.99-85 parts of type I collagen and 0.01-15 parts of type III collagen are dispersed in a solvent according to a proportion, homogenized, compounded and formed.
Preferably, the solvent is distilled water, diluted acetic acid, trifluoroacetic acid, hexafluoroisopropanol; the homogenate refers to mixing homogenate by a high-speed mixer;
more preferably, the step of homogenizing the fibrous membrane refers to: preparing homogenate with certain mass concentration; the forming step includes: adding a cross-linking agent into homogenate with a certain mass concentration to cross-link and prepare a collagen copolymer, dissolving the cross-linked copolymer into a hexafluoroisopropanol solution, placing the solution into a liquid storage pipe of a high-voltage electrostatic spinning machine, and preparing a chemical composite collagen fiber membrane through electrostatic spinning;
the homogenization step of the layered membrane refers to: preparing two homogenates with different collagen ratios; the forming step includes: evenly spreading I, III collagen homogenate with the weight ratio of 99.3:0.7 in a container, and pre-freezing the container in an ultra-low temperature refrigerator at minus 80 ℃ for 24 hours to form an outer-layer homogenate membrane; then I, III type collagen homogenate with the weight ratio of 97:3 is spread on the outer layer homogenate membrane and is pre-frozen for 24 hours at minus 80 ℃ to form a contact layer homogenate membrane, and the contact layer homogenate membrane is placed in a freezing vacuum drier at minus 50 ℃ to be dried for 24 hours to prepare a layered membrane;
the content of the type III collagen in the outer-layer homogenizing membrane is 0.01-1%, and the content of the type III collagen in the contact-layer homogenizing membrane is 1-15%.
The invention designs two forms of sponge and fibrous membrane, the fibrous membrane is divided into two forms of homogeneous component fibrous membrane and layered membrane, and can also be divided into two forms of physical compounding and chemical compounding according to a compounding mode so as to meet different requirements of wounds, wherein the chemical compounding is suitable for wound surfaces needing slow healing and adhesion prevention. The spongy and fibrous membrane is suitable for wounds with a certain area; the main purpose of the stratified membrane is to effectively increase the content of type III collagen in the contact layer, and the stratified membrane is generally used for more serious wounds.
Experimental example 1 preparation Process of layered film
(1) Evenly homogenizing I, III type collagen (w/w:99.3:0.7) with certain mass concentration, spreading in a container, pre-freezing in an ultra-low temperature refrigerator at-80 ℃ for 24h to form an outer-layer homogenate membrane;
(2) evenly spreading I, III type collagen (w/w97:3) with certain mass concentration on the outer layer even pulp membrane, pre-freezing at-80 ℃ for 24h to form a contact layer even pulp membrane, and drying in a freezing vacuum dryer at-50 ℃ for 24h to prepare the layered membrane.
The content of the type III collagen in the outer-layer homogenizing membrane is 0.01-1%, and the content of the type III collagen in the contact-layer homogenizing membrane is 1-15%.
Experimental example 2 Process for producing fiber film
(1) Weighing type I collagen and type III collagen, dispersing in distilled water according to a ratio of 93:7(W/W), mixing by a high-speed mixer to prepare homogenate with a certain mass concentration, and adding a cross-linking agent to cross-link to prepare a collagen copolymer;
(2) and dissolving the crosslinked copolymer in a hexafluoroisopropanol solution, placing the solution in a liquid storage tube of a high-voltage electrostatic spinning machine, and preparing the chemically-compounded collagen fiber membrane through electrostatic spinning.
The preparation process of the sponge-like material comprises dispersion, prefreezing and freeze-drying, and the method commonly used in the field for preparing sponges can be adopted. Mixing different collagens in proportion, homogenizing, defoaming, pre-freezing in a mould for forming, and freeze-drying.
Experimental example 3 application of the composite collagen dressing of the present invention
The composite collagen dressing and the existing single collagen material are respectively applied to the repair of the wound surface (including the diabetic foot wound surface and/or the deep wound surface) which is difficult to heal, and the composite collagen dressing can shorten the healing time of the wound surface and has higher obvious efficiency and effective rate compared with the single collagen material through verification, and the result is shown in table 2.
The composite collagen dressing according to any one of the embodiments of group 1 of the present invention, or the composite collagen auxiliary material prepared according to any one of the formulas of group 1 of table 1, can obtain similar repairing effects as described in table 2 below, and is not repeated herein.
TABLE 2 indices for wound repair
The effect is shown: the seepage is obviously reduced, the granulation tissue of the base of the wound surface is fresh, and the granulation growth is accelerated;
the method has the following advantages: the seepage is reduced and the granulation tissue grows faster.
And (4) invalidation: the seepage is not obviously reduced, and the growth of the granulation tissue at the base of the wound surface is not good.
The data are 20 experimental groups (rats purchased from the experimental animal center of Tianjin medical university), of which 16 chronic ulcer wounds and 4 deep wounds
Comparing fig. 1 and fig. 2, it can be seen that the composite collagen dressing of the present invention has more uniform pores and a more compact structure than a single collagen material.
In vitro experiments and animal experiments show that the composite collagen dressing after compounding treatment has excellent biocompatibility and histocompatibility and does not show immunogenicity. As shown in figure 3, the HE staining result of the dorsal fascia of the rat (the rat is purchased from the experimental animal center of Tianjin medical university) (the deep II-degree wound surface model repairing result) shows that a large number of new blood vessels with different diameters can be seen in the new wound surface tissues of the experimental group 14 days after the operation, and no new blood vessel is seen in the new wound surface tissues of the group A (the control group). The counting results of wound surface microvascular density Measurement (MVD) are 52.5 +/-13.8 and 75.3 +/-16.2 in sequence, and the angiogenesis amount is obviously higher than that of a control group.
Claims (11)
1. A composite collagen dressing for repairing a wound surface which is difficult to heal is characterized by being compounded by two different types of collagen;
the different types of collagen are selected from type I collagen and type III collagen;
the proportion of the type I collagen and the type III collagen is 99.99-85: 0.01-15.
2. A composite collagen dressing for healing-refractory wound repair according to claim 1, wherein said type I collagen is selected from bone, skin, bovine achilles tendon; the type III collagen is selected from human or animal skin, blood vessel wall, and amnion.
3. A composite collagen dressing for healing-refractory wound repair according to claim 2, wherein the dosage form of said composite collagen dressing is selected from the group consisting of: sponge-like materials, fibrous membranes, layered membranes.
4. The composite collagen dressing for healing-refractory wound repair according to any one of claims 1 to 3, wherein said healing-refractory wound is: the self-healing ability is poor, and the healing time is more than 2 months of wound surface.
5. A composite collagen dressing for the repair of difficult-to-heal wounds according to claim 4, characterized in that said difficult-to-heal wound is selected from the group consisting of: chronic or traumatic wound surface with difficulty in healing.
6. The preparation method of the composite collagen dressing for repairing the wound surface which is difficult to heal is characterized by comprising the following steps: dispersing type I collagen and type III collagen in a solvent in proportion, homogenizing, compounding, and molding.
7. The method for preparing a composite collagen dressing according to claim 6, comprising the steps of dispersing 99.99 to 85 parts of type I collagen and 0.01 to 15 parts of type III collagen in a solvent in proportion, homogenizing, compounding, and molding.
8. The method for preparing the composite collagen dressing according to claim 7, wherein the solvent is distilled water, diluted acetic acid, trifluoroacetic acid, hexafluoroisopropanol; the homogenization refers to mixing the homogenate by a high-speed mixer.
9. The method for preparing a composite collagen dressing according to claim 6, wherein the dosage form of the composite collagen dressing is a fibrous membrane; the homogenization step of the fiber membrane refers to: preparing homogenate with certain mass concentration; the forming step includes: adding a cross-linking agent into homogenate with certain mass concentration for cross-linking to prepare collagen copolymer, dissolving the cross-linked copolymer in a solvent, placing the solution in a liquid storage tank of a high-voltage electrostatic spinning machine, and preparing a chemical composite collagen fiber membrane by electrostatic spinning.
10. The method for preparing a composite collagen dressing as claimed in claim 6, wherein the dosage form of the composite collagen dressing is a layered film; the homogenization step of the layered membrane refers to: preparing two homogenates with different collagen ratios; the forming step includes: evenly spreading I and III collagen homogenate with the weight ratio of 99.3:0.7 in a container, and pre-freezing the container in an ultra-low temperature refrigerator at minus 80 ℃ for 24 hours to form an outer-layer homogenate membrane; then, collagen I and III homogenates with the weight ratio of 97:3 are spread on the outer-layer homogenate film and placed on a pre-freezing chamber at the temperature of minus 80 ℃ for 24 hours to form a contact-layer homogenate film, and the contact-layer homogenate film is placed on a freezing vacuum drying machine at the temperature of minus 50 ℃ for drying for 24 hours to prepare a layered film.
11. The method for preparing a composite collagen dressing as claimed in claim 10, wherein the collagen III content in the outer homogenate film is 0.01-1%, and the collagen III content in the contact layer homogenate film is 1-15%.
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| CN111991608B (en) * | 2020-08-27 | 2021-10-01 | 振德医疗用品股份有限公司 | Wound surface covering and preparation method thereof |
| CN116350537B (en) * | 2023-05-04 | 2024-03-29 | 肽源（广州）生物科技有限公司 | Collagen nanoparticle, preparation method thereof and application thereof in cosmetics |
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| CN104623642A (en) * | 2006-10-06 | 2015-05-20 | 人类起源公司 | Natural (telopeptide) placental collagen compositions |
| CN103313733A (en) * | 2010-11-15 | 2013-09-18 | 捷迈整形外科生物材料有限公司 | Bone void fillers |
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