CN1200728C - Injection-type bone morphogenetic protein using fibrin as carrier - Google Patents
Injection-type bone morphogenetic protein using fibrin as carrier Download PDFInfo
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- CN1200728C CN1200728C CN 02114506 CN02114506A CN1200728C CN 1200728 C CN1200728 C CN 1200728C CN 02114506 CN02114506 CN 02114506 CN 02114506 A CN02114506 A CN 02114506A CN 1200728 C CN1200728 C CN 1200728C
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Abstract
The present invention discloses injected bone morphogenetic protein using fibrin as carriers, which is suitable for clinically treating fractures, nonunion and bone defect. Fibrin separated from patients' blood or healthy blood donors' blood is used as carriers, compounded cow bone morphogenetic protein is directly injected to the local injury part percutaneously to perform the functions of slow release of the carriers, and new bone growth promotion, fracture union and wound repair acceleration of the bone morphogenetic protein. The present invention does not result in immunogenicity or rejection reaction, and has no adverse reaction after implanted in the body. Before used, the fibrin is liquid, can be rapidly solidified in the body, and can be shaped randomly. In addition, the functions of hemostasis and cementation of the fibrin are favourable for preventing and curing local hematoma, which provides a favorable local environment for the regeneration and the union of bones. Animal experiments and clinical tests prove that the present invention has exact treatment effect, wide range of application, easy operation, safety and reliability.
Description
One, affiliated field
The present invention relates to a kind of protein injection agent of medical domain, particularly a kind ofly be used for disease treatments such as clinical fracture, bone does not connect, bone be damaged and can inject the bone morphogenetic protein of use.
Two, background technology
The main effect of bone morphogenetic protein is to promote that undifferentiated mesenchymal cell and bone are raising of cell and to osteoblast differentiation.Because it has bone-inducting active efficiently, therefore promote union of fracture, the treatment bone is damaged and bone does not connect aspect fabulous potential applicability in clinical practice is arranged.The method in clinical use of having now mostly is bone morphogenetic protein is compounded on the solid material, implant wound site, complicated operation, cost height by operation, restricted application does not especially need the case of operative treatment to be difficult to use to the maximum closed fracture of clinical appearance and other.
Three, summary of the invention
The defective or the deficiency that partly exist according to above-mentioned prior art, the objective of the invention is, a kind of damaged patient of bone does not connect, bone who needing not only to can be used for operative treatment is provided, can also uses ordinary syringe percutaneous direct injection, thereby available all patients with bone injury.Having human fiber's albumen that the scope of application is extensive, easy and simple to handle, therapeutic effect is good is the injection-type bone morphogenetic protein of carrier.
To achieve these goals, the technical conceive that the present invention adopts is, use blood samples of patients or blood bank's healthy adult human blood, prepare the carrier of Fibrinogen fast by the low-temperature centrifugation method, be converted into fibrin clot after the effect by thrombin and aprotinin during use makes Fibrinogen/bone morphogenetic protein complex in being expelled to body as bone morphogenetic protein.Utilize fibrin clot bone morphogenetic protein to be slowly released by the characteristic that body slowly absorbs, thereby make bone morphogenetic protein all bring into play its biological action, reach the effect of inducing new bone formation and promoting repair in trauma in the overall process of repair in trauma.
The technical scheme that is adopted is:
With the fibrin is the injection-type bone morphogenetic protein of carrier, according to the following steps preparation:
1) obtains the preceding aseptic patient's of the taking autogenous vein blood of health adult's anticoagulation or treatment from hospital blood bank/blood station;
2) centrifugal 20 minutes separated plasmas of 3000RPM, refrigerated centrifugation extracts Fibrinogen, adds the Bovine Bone Morphoge Protnetin extract, and ratio is (2～5) mg: 1mg (Fibrinogen: bone morphogenetic protein); It is standby that floccule is made in said mixture-40 a ℃ lyophilizing;
3) Fibrinogen/bone morphogenetic protein mixture being made into concentration with normal saline is that the suspension of 4mg/ml～8mg/ml is as composition A; With the calcium chloride solution compound concentration is that 250IU/ml～400IU/ml thrombin solution is composition B;
4) adding concentration in composition B is the aprotinin of 0.5mg/ml～50mg/ml.
Another characteristics of the present invention are, described Bovine Bone Morphoge Protnetin extract, it is the artificial according to a conventional method multiple bone morphogenetic protein mixture that extracts from the fresh cortical bone of calf, key step is defat, deproteinization, guanidine hydrochloride, urea extraction, room temperature is dry down, has bone-inducting active through mice flesh bag experiment confirm.
The present invention utilizes fibrinogenic dissolution characteristics to can be made into injection, thereby needing not only to can be used for the damaged patient of bone does not connect, bone of operative treatment, can also use ordinary syringe percutaneous direct injection, thereby can be used for all patients with bone injury.Add respectively in two tubes of dual barrel syringe with isopyknic composition A and composition B during use, mixing gets final product in the process of injection.The present invention has advantages such as the scope of application is extensive, easy and simple to handle, therapeutic effect is definite.From human blood, extract fibrin and be used further to human body, do not have immunogenicity and rejection, have no adverse reaction after implanting.Especially before treatment, extract the propagation that patient's autoblood can also be avoided hematologic disease.Owing to only use the plasma fraction in the blood, other can feed back to the patient after becoming divisional processing, thereby patient's blood system is not had harmful effect.Be liquid before fibrin uses, can solidify rapidly in vivo, arbitrarily plastotype.Simultaneously, fibrinous hemostasis, bonding effect help preventing and treating the formation of local hematoma, for the regeneration and the healing of bone provides good local environment.
Four, the specific embodiment
The present invention is described in further detail below in conjunction with embodiment.
Embodiment 1: by technical scheme of the present invention, is the injection-type bone morphogenetic protein of carrier with the fibrin, and preparation according to the following steps:
1) obtains the preceding aseptic patient's of the taking autogenous vein blood of health adult's anticoagulation or treatment from hospital blood bank/blood station;
2) centrifugal 20 minutes separated plasmas of 3000RPM, refrigerated centrifugation extracts Fibrinogen, adds the Bovine Bone Morphoge Protnetin extract, and ratio is 2mg: a 1mg (Fibrinogen: bone morphogenetic protein); It is standby that floccule is made in said mixture-40 a ℃ lyophilizing;
The Bovine Bone Morphoge Protnetin extract, be the artificial according to a conventional method multiple bone morphogenetic protein mixture that extracts from the fresh cortical bone of calf, key step is defat, deproteinization, guanidine hydrochloride, urea extraction, room temperature is dry down, has bone-inducting active through mice flesh bag experiment confirm;
3) Fibrinogen/bone morphogenetic protein mixture being made into concentration with normal saline is that the suspension of 8mg/ml is as composition A; With the calcium chloride solution compound concentration is that the 250IU/ml thrombin solution is composition B;
4) adding concentration in composition B is the aprotinin of 50mg/ml.
Embodiment 2: they by technical scheme of the present invention, are the injection-type bone morphogenetic protein of carrier with the fibrin, and preparation according to the following steps:
1) obtains the preceding aseptic patient's of the taking autogenous vein blood of health adult's anticoagulation or treatment from hospital blood bank/blood station;
2) centrifugal 20 minutes separated plasmas of 3000RPM, refrigerated centrifugation extracts Fibrinogen, adds the Bovine Bone Morphoge Protnetin extract, and ratio is 4mg: a 1mg (Fibrinogen: bone morphogenetic protein); It is standby that floccule is made in said mixture-40 a ℃ lyophilizing;
The Bovine Bone Morphoge Protnetin extract, be the artificial according to a conventional method multiple bone morphogenetic protein mixture that extracts from the fresh cortical bone of calf, key step is defat, deproteinization, guanidine hydrochloride, urea extraction, room temperature is dry down, has bone-inducting active through mice flesh bag experiment confirm;
3) Fibrinogen/bone morphogenetic protein mixture being made into concentration with normal saline is that the suspension of 4mg/ml is as composition A; With the calcium chloride solution compound concentration is that the 300IU/ml thrombin solution is composition B;
4) adding concentration in composition B is the aprotinin of 30mg/ml.
Embodiment 3: they by technical scheme of the present invention, are the injection-type bone morphogenetic protein of carrier with the fibrin, and preparation according to the following steps:
1) obtains the preceding aseptic patient's of the taking autogenous vein blood of health adult's anticoagulation or treatment from hospital blood bank/blood station;
2) centrifugal 20 minutes separated plasmas of 3000RPM, refrigerated centrifugation extracts Fibrinogen, adds the Bovine Bone Morphoge Protnetin extract, and ratio is 5mg: a 1mg (Fibrinogen: bone morphogenetic protein); It is standby that floccule is made in said mixture-40 a ℃ lyophilizing;
The Bovine Bone Morphoge Protnetin extract, be the artificial according to a conventional method multiple bone morphogenetic protein mixture that extracts from the fresh cortical bone of calf, key step is defat, deproteinization, guanidine hydrochloride, urea extraction, room temperature is dry down, has bone-inducting active through mice flesh bag experiment confirm;
3) Fibrinogen/bone morphogenetic protein mixture being made into concentration with normal saline is that the suspension of 6mg/ml is as composition A; With the calcium chloride solution compound concentration is that the 400IU/ml thrombin solution is composition B;
4) adding concentration in composition B is the aprotinin of 0.5mg/ml.
Embodiment 4: they by technical scheme of the present invention, are the injection-type bone morphogenetic protein of carrier with the fibrin, and preparation according to the following steps:
1) obtains the preceding aseptic patient's of the taking autogenous vein blood of health adult's anticoagulation or treatment from hospital blood bank/blood station;
2) centrifugal 20 minutes separated plasmas of 3000RPM, refrigerated centrifugation extracts Fibrinogen, adds the Bovine Bone Morphoge Protnetin extract, and ratio is 3mg: a 1mg (Fibrinogen: bone morphogenetic protein); It is standby that floccule is made in said mixture-40 a ℃ lyophilizing;
The Bovine Bone Morphoge Protnetin extract, be the artificial according to a conventional method multiple bone morphogenetic protein mixture that extracts from the fresh cortical bone of calf, key step is defat, deproteinization, guanidine hydrochloride, urea extraction, room temperature is dry down, has bone-inducting active through mice flesh bag experiment confirm;
3) Fibrinogen/bone morphogenetic protein mixture being made into concentration with normal saline is that the suspension of 6mg/ml is as composition A; With the calcium chloride solution compound concentration is that the 350IU/ml thrombin solution is composition B;
4) adding concentration in composition B is the aprotinin of 15mg/ml.
Above-mentioned calcium chloride solution compound concentration can be adjusted between 250IU/ml～400IU/ml, and the aprotinin concentration that adds among the B is adjusted between 0.5mg/ml～50mg/ml, can both reach purpose of the present invention.
Instantiation 1: fracture, fracture surgery bone does not connect patient is as follows with this repair materials therapeutic process: manual reduction, observe under the TV X-ray machine perspective fracture end para-position satisfied after, get this material 10ml, percutaneous injection is to fracture site under the perspective.Gypsum Fibrosum or external fixer are fixed, and the 4 visible union of fracture in week back are good.Show this material accelerating bone healing speed effectively, can avoid operation for the second time for the treatment of bone does not connect.
Instantiation 2: the vertebral compression fracture patient is venous blood samples liquid before operation, make fibrin, and compound bone morphogenetic protein is standby.With the reduction of the fracture or strut the vertebral body of compression, use internal fixation by operation, with fibrin/bone morphogenetic protein of preparing before the art be expelled to vertebral body after composition B mixes, conventional method is closed wound.Autohemic cell component can feed back in operation process gives the patient with losing blood in the compensation operation.In this type of trauma care, this material not only can promote union of fracture, can also induce new bone formation in vertebral body inside, recovers original form of vertebral body and mechanical strength.
Syringe is a special designs, disposable use.Feature is: 1. the bitubular is parallel, and volume is 5ml, and two propeller rear ends link to each other, and uses one to advance handle.2. two exit passages are spiral type, converge in end, are convenient to two kinds of composition mix homogeneously.3. 3 at external syringe needle, fine needle head are No. 12 common syringe needles of steel, use for percutaneous injection; Thick needle is special hour hand head, and internal diameter 1.8mm is for using in the operation; Shower nozzle is special, can make atomisation behind the liquid mixing.
Add an amount of aprotinin among the composition B, reach the purpose of slowing down the fibrin infiltration rate.
The scope of application of the present invention has orthopedics, neurosurgery, oral surgery etc.
Before using equal-volume composition A and composition B are added respectively in two tubes of dual barrel syringe, external special-purpose syringe needle, percutaneous is injected directly into the damage part under the guiding of TV X-ray machine.
Advantage of the present invention is: promote union, accelerated bone forms and the knitting definite effect; Use letter Just, indication is extensive; Histocompatbility is good, and is safe and reliable.
Claims (2)
1. one kind is the preparation method of the injection-type bone morphogenetic protein of carrier with the fibrin, it is characterized in that, carries out according to the following steps:
1) obtains the preceding aseptic patient's of the taking autogenous vein blood of health adult's anticoagulation or treatment from hospital blood bank/blood station;
2) extract Fibrinogen after the separated plasma, the adding mass ratio is that the Bovine Bone Morphoge Protnetin extract of 2～5: 1 dosage is standby;
3) Fibrinogen/Bovine Bone Morphoge Protnetin mixture is mixed with normal saline, the suspension that is made into mixture concentration and is 4mg/ml～8mg/ml is as composition A; With the calcium chloride solution compound concentration is that 250IU/ml～400IU/ml thrombin solution is composition B; Composition A does not mix external with composition B;
4) adding concentration in composition B is the aprotinin of 0.5mg/ml～50mg/ml.
2. according to claim 1 is the preparation method of the injection-type bone morphogenetic protein of carrier with the fibrin, it is characterized in that: described Bovine Bone Morphoge Protnetin extract, it is the artificial multiple bone morphogenetic protein mixture that extracts from the fresh cortical bone of calf, extracting method is: defat, deproteinization, guanidine hydrochloride, urea extraction have bone-inducting active through mice flesh bag experiment confirm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN 02114506 CN1200728C (en) | 2002-04-09 | 2002-04-09 | Injection-type bone morphogenetic protein using fibrin as carrier |
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| CN 02114506 CN1200728C (en) | 2002-04-09 | 2002-04-09 | Injection-type bone morphogenetic protein using fibrin as carrier |
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| CN1376514A CN1376514A (en) | 2002-10-30 |
| CN1200728C true CN1200728C (en) | 2005-05-11 |
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| CN106389393A (en) * | 2016-11-23 | 2017-02-15 | 中国人民解放军第三军医大学第附属医院 | Biological protein glue-triamcinolone acetonide slow release agent as well as preparation method and application of slow release agent |
| CN106474550A (en) * | 2016-12-02 | 2017-03-08 | 中国人民解放军第八十八医院 | A kind of construction method of tendon osseous tissue bone sex camplex |
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