WO2017101024A1

WO2017101024A1 - Modified biomedical material product

Info

Publication number: WO2017101024A1
Application number: PCT/CN2015/097455
Authority: WO
Inventors: 高敏
Original assignee: 江阴市本特塞缪森生命科学研究院有限公司
Priority date: 2015-12-15
Filing date: 2015-12-15
Publication date: 2017-06-22

Abstract

A product formed of a mussel adhesive protein and a biomedical material, relating to the products formed of a mussel adhesive protein and materials for repairing the musculoskeletal system such as joint and tendon, soft tissue materials for skin, breast, esophagus, respiratory tract, bladder and the like, cardiovascular system materials such as artificial heart valves, blood vessels, and cardiovascular cannulas, medical membrane materials such as blood purification membranes and separation membranes, gas selective permeable membranes, and corneal contact lens, and materials for clinical diagnosis and biosensor and the like. On the basis of the original functions of the biomedical material, the product is more advantageous for rebuilding or recovering the damaged human tissues or organs, having stronger physical and chemical stability, extending the degradation period of proteins, being more advantageous for the exhibition of the bioactivity of Mussel adhesive protein.

Description

An improved biomedical material product

Technical field

The present invention relates generally to the field of medical product technology and, more particularly, to an improved biomedical material product for curing mussel mucin on biomedical materials.

Background technique

Biomedical materials are materials used to diagnose, treat, repair, or replace a diseased tissue, organ, or function of an organism. It is the basis for the study of artificial organs and medical devices and has become an important branch of contemporary materials science. Especially with the vigorous development and major breakthroughs in biotechnology, biomedical materials have become a hotspot for scientists and scientists in various countries to compete for research and development.

Biomedical materials can be divided into bone-muscle system repair materials such as bones, joints and tendons according to their uses, soft tissue materials such as skin, breast, esophagus, respiratory tract and bladder, artificial heart valves, blood vessels, cardiovascular intubation and other cardiovascular system materials. Blood purification membranes and separation membranes, gas selective permeable membranes, contact lens and other medical membrane materials, clinical diagnostics and biosensor materials. At present, the focus of biomedical materials research is to find biomedical materials with better tissue compatibility, biodegradability, corrosion resistance, durability and versatility under the premise of ensuring safety.

Mussel adhesive protein (MAP), also known as Mytilus edulis foot protein (Mefp), is a marine shellfish, Mytilus edulis Linnaeus, and a small shell mussel (Mytilus coruscus). A special protein secreted by Perna viridis. Mussels are usually attached in groups to the reefs on the coast or to the bottom of the ship, and have the ability to withstand wave impacts in the offshore. In fact, mussels can be attached extremely strongly to the substrate of any material, such as metal, wood, glass, and the like. The main reason for the above characteristics of mussels is that they can form and store this special mucin in the girth of the foot. The mussels release the mucin through the foot silk to a solid surface such as rock to form a water-resistant combination. Fix yourself.

Currently, 11 mucin subclasses have been identified from mussels, including mefp1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, collagen pre-COL-P, pre-COL-D. , pre-COL-NG, foot silk matrix proteins PTMP and DTMP (Zhu Xi et al, Advances in Marine Science, 2014, 32(4): 560-568). Mussel mucin has two structural features: (1) containing lysine, which has a high loading of positive charge; (2) containing 3,4 dihydroxyphenylalanine (DOPA, dopa). Human cells and groups Weave with a negative charge. Mussel mucin plays a protective and therapeutic role by tightly binding cells and tissues through the electrostatic interaction between its own positive charge and the negative charge of cells and tissues. In addition, dopa oxidation produces ortho-dioxins, which can be cross-linked with unoxidized dopa to form a membrane or a network scaffold, which promotes the protein to adhere more closely and firmly to the surface of the human body, thereby protecting. Mussel mucin is a macromolecular protein that is completely degraded in the human body for about 3-10 days. Its ability to attach to cell tissues is excellent, so that mussel mucin can be stabilized locally and continue to function.

At present, the application fields of mussel mucin products are very limited. Commercial mussel mucin products include Cell-Tak from BD Biosciences, MAP Trix from Kollodis, Korea, and Hydrogel from Biopolymer, Sweden. These products are either used directly in the mussel mucin solution state, or are stored as lyophilized powder formulations and dissolved prior to use. Their primary application is limited to microscopic cell adhesion and tissue adhesives. Mussel mucin has also been reported for use in membrane repair, as a coating against seawater corrosion.

Mussel mucin can be used for wound healing process because it has the function of promoting cell adhesion and crawling, but its application in biomedical materials has not been reported.

Summary of the invention

It is an object of the present invention to provide an improved biomedical material product comprising mussel mucin and a biomedical material wherein the mussel mucin is cured on the biomedical material in a uniform coverage.

Mussel mucin used herein refers to Mytilus edulis Linnaeus, Mytilus coruscus or Perna viridis from the Mytilidae bivalve mollusc. 11 subclasses of mussel mucin, currently known as purified from marine mussels: mefp1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, collagen pre-COL- A mixture of one or more of P, pre-COL-D, pre-COL-NG, foot silk matrix proteins PTMP and DTMP. The mussel viscosity of the mussel mucin used herein may have a pH of 1.0 to 7.0 in an aqueous solution, and particularly may be in the range of pH 3.0 to 6.5 to make the effect better.

The mussel mucin used herein may also be obtained by a method of biosynthesis, comprising a mixture of one or more of the known 11 mussel mucin subclasses. The artificially biosynthesized mussel mucin used herein may have a pH of 1.0 to 7.0 in aqueous solution, and particularly may be in the range of pH 3.0 to 6.5 to make it more effective.

The mussel mucin used herein may also be a hydrolyzed peptide obtained by hydrolyzing mussel mucin from a natural source or an artificial biosynthesis, or a synthetic peptide containing a functional group obtained by artificial synthesis. The mussel mucin hydrolyzed peptide or synthetic peptide used herein may have a pH of 1.0 to 7.0 in an aqueous solution, and particularly may be in the range of pH 3.0 to 6.5 to make the effect better.

The mussel mucin used herein can be obtained by the following preparation method, for example, a method for separating and purifying mussel mucin using mixed adsorption chromatography in Chinese Patent No. ZL200710179491.0, a kind of carboxy using Chinese Patent No. ZL200710179492.5 A method for purifying mussel mucin by methyl ion exchange chromatography, a method for separating and purifying mussel mucin using salting out and dialysis, Chinese Patent No. ZL200910087567.6.

The mussel mucin used herein may be in the form of a solution or a lyophilized powder.

In the context of the present invention, a biomedical material refers to a biomedical material other than a wound closure material, a dressing, a bone implant, or a medical stent, which may be an artificial joint and a joint prosthesis, a patch, a waterproof tape, an artificial blood vessel. And endovascular prosthesis, artificial heart valve, artificial ear implant, artificial organ prosthesis, artificial soft tissue filling, artificial amniotic membrane, intervertebral disc prosthesis, spinous process implant, artificial heart valve, blood vessel, cardiovascular intubation, etc. Cardiovascular materials, blood purification membranes and separation membranes, gas selective permeable membranes, contact lens and other medical membrane materials, drug release carrier materials, clinical diagnostics and biosensor materials.

The invention provides a method for preparing an improved biomedical material, comprising the steps of:

(1) preparing a mussel mucin solution,

(2) Curing the mussel mucin solution to a biomedical matrix material to obtain an improved biomedical material.

The term "curing" as used in the present invention refers to the adsorption of mussel mucin to a biomedical material by physical means.

In one embodiment, the mussel mucin solution can be added to the biomedical matrix material for a period of time to cure the mussel mucin to the biomedical matrix material.

In one embodiment, the mussel mucin solution can be sprayed onto the surface of the biomedical matrix material for a period of time to cure the mussel mucin to the biomedical matrix material.

In one embodiment, the biomedical matrix material can be infiltrated into the mussel mucin solution for a period of time to cure the mussel mucin to the biomedical matrix material.

In an embodiment of the invention, the mussel mucin solution may be acidic, neutral or weakly alkaline The solution is prepared and optimally treated with an acidic solution.

In the embodiment of the present invention, the mussel mucin solution may be formulated from citric acid, acetic acid, sodium citrate, oxalic acid, carbonic acid, phosphoric acid, benzoic acid, sodium carbonate, or the like, but is not limited to the above solvent.

In an embodiment of the invention, the concentration of mussel mucin may be from 0.01 to 150 mg/ml, in particular the concentration of mussel mucin in the product may be from 0.1 to 15 mg/ml.

In an embodiment of the invention, the mussel mucin solution is used in an amount sufficient to uniformly cover or infiltrate the biomedical material.

In an embodiment of the invention, the temperature for curing may be any, preferably 10-40 °C.

In an embodiment of the invention, the period of time for curing may be any, preferably from 10 to 60 minutes.

The present invention provides an artificial joint and a joint prosthesis in which mussel mucin is cured onto an existing artificial joint material. Artificial joints and joint prostheses may include: hip joints, knee joints, shoulder joints, elbow joints, knuckles, ankle joints, and the like, and prostheses thereof.

The present invention provides a patch in which mussel mucin is cured onto an existing patch material. Patches may include: dura mater patch, vascular patch, vaginal patch, visceral patch, sputum patch, soft tissue patch, and the like.

The present invention provides an artificial blood vessel and an endovascular prosthesis in which mussel mucin is cured onto an existing vascular material. The artificial blood vessel may include: a polytetrafluoroethylene vascular graft, a double velvet woven artificial blood vessel, a polyester artificial blood vessel, an expanded polytetrafluoroethylene artificial blood vessel; the endovascular prosthesis may include: a thoracic aortic stent system, an aortic stent system, abdomen Aortic and radial artery stent systems.

The present invention provides a prosthetic heart valve in which mussel mucin is cured onto an existing prosthetic heart valve material. Prosthetic heart valves can include: bio heart valves, mechanical heart valves, transcatheter implantable heart valves, heart valve forming rings, and the like.

The present invention provides an artificial ear implant in which mussel mucin is cured onto an existing artificial ear implant material. Artificial ear implants may include: cochlear implants, ossicular prostheses, and the like.

The present invention provides an artificial organ prosthesis in which mussel mucin is cured onto an existing artificial prosthetic material. Artificial organ prostheses may include: breast prosthesis, prosthetic eye, esophagus, urethra, kidney, heart, larynx, cornea, tendon, anal occluder, penile prosthesis, tympanoplasty prosthesis, patella prosthesis, Nasal prosthesis, facial prosthesis, ear prosthesis, subcutaneous soft tissue implant, intervertebral disc prosthesis, and the like.

The present invention provides an artificial soft tissue filler in which mussel mucin is cured onto an existing soft tissue filler material. Artificial soft tissue fillers can include: bioabsorbable fillers, polyester fillers, polypropylene fillers, and the like.

The present invention provides an artificial amniotic membrane in which mussel mucin is cured onto an existing artificial amniotic material.

The present invention provides a spinous process implant in which mussel mucin is cured onto existing spinous implant material. Spinous process implants can include: interspinous dynamic stabilization systems, spinal surgical implants - percutaneous interspinous distraction systems, vertebral column blocks, and the like.

The definition of the related biomedical material product described in the present invention is derived from the "Medical Device Classification Catalogue" (2013 edition) promulgated by the State Food and Drug Administration. All of the products of the present invention can be prepared by the above-described method of preparing an improved biomedical material.

The invention provides an improved biomedical material capable of exerting the activity of mussel mucin on a biomedical material, imparting biological structure and biological function to the biomedical material, so as to mobilize in the body and exert self-repair and perfection of the body. Ability is more conducive to reconstruction or rehabilitation of damaged human tissues or organs. In particular, when mussel mucin is combined with biomedical materials, the degree of cross-linking of mussel mucin is increased, the physical and chemical stability of the protein is enhanced, the degradation cycle of the protein is prolonged, and the mussel mucinous organism is more favorable. The play of activity.

detailed description

The invention will now be further described in conjunction with specific embodiments. It should be noted that the biomedical material product cured with mussel mucin of the present invention can be prepared by the above method, and the effects have been tested and verified within the scope of the present invention, hereinafter, only for explanation, only in A few of the examples are described in the examples, which should not be construed as limiting the invention. The nature and advantages of the products of the present invention are further illustrated in the following non-limiting examples.

Example 1: Mussel mucin solidified to artificial joints to promote rehabilitation

Mussel mucin solidified artificial joint preparation: take 20ml of mussel mucin solution of 0.5mg/ml, spray mussel mucin to the surface of artificial knee joint (titanium alloy, Nanjing Honghe Precision Forging Co., Ltd.), keep at 30 °C After a minute, dry, take it out and set aside.

Six cases of osteoarthritis were collected, and patients with artificial knee joints were replaced and confirmed by an orthopaedic surgeon. The knee joints of the affected area were replaced with untreated artificial joints, joints that sprayed mussel mucin before use, and joints that cured mussel mucin. All patients were able to lie flat on the bed on the first day after surgery, and they could sit on the second day. On the third day, they could walk slowly and kneel down. Patients with artificial knee joints with cured mussel mucin can abandon their walking and return to normal life 30 days after surgery; patients with artificial knee joints who spray mussel mucin before use will return to normal life 40 days after surgery; Patients with untreated knee joints returned to normal life 50 days after surgery. It can be seen that artificial knee joint patients who cure mussel mucin recover faster.

Example 2: Mussel mucin solidified to the patch to promote endothelial cell growth

Preparation of mussel mucin-cured xenon patch: Take 0.1ml/ml mussel mucin solution 20ml, soak the xenon patch (Lappu medical polypropylene mesh plug, Wuhan Lanpu Medical Products Co., Ltd.) in 贻The shellfish protein solution was kept at room temperature for 30 minutes, and then dried at 45 ° C for use.

Two patients with hernia were enrolled and confirmed by a general practitioner. The untreated patch and mussel mucin cured patch were used. The operation was performed by a laparoscopic double-layer repair device. In patients with patch-cured mussel mucin, the surface of the patch has been covered by the body's own endothelial cells 1 month after surgery, and the patch has been completely embedded by endothelial cells until 3 months. In patients who had not been treated with mussel mucin, endothelial cells did not completely cover the surface of the patch 1 month after surgery. By 4 months after surgery, the patch was completely embedded in endothelial cells. It can be seen that the patch of the mussel mucin is more favorable for the coverage of endothelial cells and reduces the recurrence of hernia.

Example 3: Mussel mucin solidified to artificial blood vessels to promote vascular membrane growth

Preparation of mussel mucin-cured artificial blood vessel: Take 10ml of mussel mucin solution of 1.5mg/ml, soak the artificial blood vessel (Bard Medical Devices Co., Ltd.) in mussel mucus, keep it at room temperature for 15 minutes, take it out and dry it. use.

Six male New Zealand white rabbits weighing 4.0-4.5 kg were taken, and the blood vessels of 2 cm in length were taken from the heart artery, and artificial blood vessels, artificial blood vessels directly sprayed with mussel mucin, and artificial blood vessels that cured mussel mucin were implanted into the blood vessels. Vascular defects, 2 in each group. One animal implanted directly into the artificial blood vessel died on the second day after surgery, and the anatomical results showed local oozing of the implanted artificial blood vessel. The remaining animals were sacrificed 1 month after surgery and the healing of the implanted artificial blood vessels was observed. The vascular suture at the site of implantation of the animal directly implanted with the artificial blood vessel was substantially healed, and neither the intravascular nor the adventitia were formed. Mussel mucin The artificial blood vessel was sprayed, the suture was healed, the blood vessel and the outer membrane were not completely formed, and a small amount of thrombus was formed on the inner wall of the blood vessel. The artificial blood vessel which cures mussel mucin is used, the suture is healed, the inner and outer vessels are formed, and no thrombus is formed on the inner wall of the blood vessel. It can be seen that the artificial blood vessel that cures mussel mucin is more favorable for the formation of intravascular and adventitial membranes and to avoid the formation of thrombus.

Claims

An improved biomedical material consisting of mussel mucin and biomedical materials, wherein mussel mucin is cured on a biomedical material in a uniform coverage.
The improved biomedical material according to claim 1 wherein said mussel mucin is from a subclass: mefp1, mefp-2, mefp-3, mefp-4, mefp-5, mefp-6, collagen pre-COL- A mixture of one or more of P, pre-COL-D, pre-COL-NG, foot silk matrix proteins PTMP and DTMP.
The improved biomedical material of claim 1 wherein the biomedical material is a biomedical material other than a wound closure material, a dressing, a bone implant, a medical stent, including an artificial joint and a joint prosthesis, including a hip joint, a knee joint, Shoulder joint, elbow joint, knuckle joint, ankle joint and other prosthesis, patch, including dura mater patch, vascular patch, vaginal patch, visceral patch, sputum patch, soft tissue patch, waterproof tape, including Fabric tape, polyethylene tape, paper tape, silicone tape, artificial blood vessel and endovascular prosthesis including polytetrafluoroethylene vascular graft, double velvet woven artificial blood vessel, polyester artificial blood vessel, expanded polytetrafluoroethylene artificial blood vessel; chest Aortic stent system, aortic stent system, endovascular prosthesis of abdominal aorta and radial artery stent system, artificial heart valve, including biological heart valve, mechanical heart valve, transcatheterized heart valve, heart valve forming ring, artificial Ear implants, including cochlear implants, auditory small bone prostheses, artificial organ prostheses, including breast prostheses, prosthetic eyes, esophagus, urethra , kidney, heart, larynx, cornea, tendon, anal occluder, penile prosthesis, tympanoplasty prosthesis, humeral prosthesis, nasal prosthesis, facial prosthesis, ear prosthesis, subcutaneous soft tissue Inclusion, intervertebral disc prosthesis, artificial soft tissue filler, including bioabsorbable filler, polyester filler, polypropylene filler, artificial amniotic membrane, intervertebral disc prosthesis, spinous process implant, including interspinous dynamic stabilization system, spine Surgical implants - percutaneous interspinous distraction system, vertebral column blocks, artificial heart valves, blood vessels, cardiovascular intubation and other cardiovascular system materials, blood purification membranes and separation membranes, gas selective permeable membranes, corneal contact Mirror and other medical membrane materials, drug release carrier materials, clinical diagnostics and biosensor materials.
A method of making an improved biomedical material comprising:

(1) preparing a mussel mucin solution,

(2) Curing the mussel mucin solution to a biomedical matrix material to obtain an improved biomedical material.
A method according to claim 4, wherein said curing is by adding a mussel mucin solution to the biomedical matrix material, or spraying the surface of the biomedical matrix material, or infiltrating the biomedical matrix material into the mussel mucin solution. Mussel mucin is cured to the biomedical matrix material after a period of time.
A method according to claim 4 wherein the mussel mucin solution is formulated from an acidic, neutral or weakly alkaline solution, especially from an acidic solution.
The method of claim 6 wherein the mussel mucin solution is formulated from citric acid, acetic acid, sodium citrate, oxalic acid, carbonic acid, phosphoric acid, benzoic acid, sodium carbonate.
A method according to any one of claims 4-7, wherein the mussel mucin concentration is from 0.01 to 150 mg/ml, in particular from 0.1 to 15 mg/ml.
A method according to any one of claims 4 to 7, wherein the curing temperature is from 10 to 40 °C.
A method according to any one of claims 4 to 7, wherein the curing time is from 10 to 60 minutes.
An artificial joint and a joint prosthesis in which mussel mucin is solidified on the artificial joint and the joint prosthesis in a uniform covering manner.
A patch in which mussel mucin is cured in a uniform coverage on the patch.
A waterproof tape in which mussel mucin is cured on a blanket in a uniform coverage.
An artificial blood vessel and an endovascular prosthesis in which mussel mucin is solidified on an artificial blood vessel or an endovascular prosthesis in a uniform covering manner.
An artificial heart valve in which mussel mucin is cured in a uniform coverage on a prosthetic heart valve.
An artificial ear implant in which mussel mucin is cured on a artificial ear implant in a uniform coverage.
An artificial organ prosthesis in which mussel mucin is cured in a uniform coverage on an artificial organ prosthesis.
An artificial soft tissue filler in which mussel mucin is cured in a uniform coverage on a soft tissue filling.
An artificial amniotic membrane in which mussel mucin is cured in a uniform coverage on an artificial amniotic membrane.
A spinous process implant in which mussel mucin is cured in a uniform coverage on a spinous process implant.