CN107583108A - A kind of preparation method of bone grafting material - Google Patents
A kind of preparation method of bone grafting material Download PDFInfo
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- CN107583108A CN107583108A CN201711054017.5A CN201711054017A CN107583108A CN 107583108 A CN107583108 A CN 107583108A CN 201711054017 A CN201711054017 A CN 201711054017A CN 107583108 A CN107583108 A CN 107583108A
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Abstract
The invention discloses a kind of preparation method of bone grafting material：By ox bone, sheep bone or pig bone in weeding out soft tissue, deionized water rinsing is used under aseptic condition；Ox bone, sheep bone or pig bone are soaked using liquor natrii hypochloritis, finally rinse bone tissue surface repeatedly with deionized water；Ox bone, sheep bone or pig bone are shredded, obtained skeletal grain soaked using deionized water, then centrifugal treating is carried out to skeletal grain；Skeletal grain after processing is immersed in ether solution at room temperature and carries out ungrease treatment, is treated after the completion of ungrease treatment with deionized water rinsing skeletal grain to remove ether；Obtained skeletal grain is subjected to freezing processing；Obtained skeletal grain is dipped into trypsin solution, carries out ferment treatment；To obtained skeletal grain deionized water rinsing, it is dried in vacuo afterwards, then dried skeletal grain is subjected to sterilization treatment.The preparation method of the present invention, the bone grafting material produced can be efficiently applied to bone defect healing.
Description
Technical field
The invention belongs to medical material tech field, and in particular to a kind of preparation method of bone grafting material.
Background technology
In recent years, with the sustainable growth of motor vehicle number, frequent accidents occur on road surface, and by major motor vehicle
The disease such as fracture, Cranial defect is more and more caused by accident, pollution or the fracture infected, Cranial defect be present for handling
Etc. being problem that current orthopedist faces.
In traditional method, two phase cures are generally adopted by, first phase treatment mainly cleaning wound face, prevent from feeling
Dye, after the possibility of infection to be confirmed is brought under control, carrying out second Bone Defect Repari and Fracture internal fixaiion.The defects of conventional method
It is：Treatment cycle is very long, and patient can be very painful, and patient's long-term bed can also cause other problemses.In fact, Cranial defect
Repair the important topic that always electrosurgical operation person constantly studies.
Although medical science is constantly developing, still it is used for bone without good replacement for large-scale Cranial defect
The reparation and treatment of defect.From the point of view of research at this stage：There is nature bone derived material preferable osteoacusis and self-bone grafting to live
Property, bone- xenograft derived material has rejection clearly compared with homogeneous allogenic bone derived material；In addition, allogeneic
Bone derived material and autologous bone are closer, therefore as the preferred raw materials in bone defect healing.
At present, existing homogeneous allogenic bone derived material is to be obtained from people's bone tissue after processing, and source is relatively fewer,
Cost of manufacture is high；In addition, it is impossible to ensure the quality of bone tissue, just in case human body has infectious disease etc., entail dangers to bone defect healing
The life of person.As can be seen here, it is highly desirable to develop a kind of homogeneous allogenic bone derived material that can overcome drawbacks described above.
The content of the invention
It is an object of the invention to provide a kind of preparation method of bone grafting material, preparation method is simple, the bone produced
Graft materials can be efficiently applied to bone defect healing.
The technical solution adopted in the present invention is a kind of preparation method of bone grafting material, specifically real according to following steps
Apply：
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, rushed under aseptic condition with deionized water
Wash so that bone tissue does not show color；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, are killed potential
Virus, finally rinse bone tissue surface repeatedly with deionized water；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, will obtain
Skeletal grain using deionized water soak, then to skeletal grain carry out centrifugal treating remove impurity therein；
Step 4, the skeletal grain after step 3 is handled is immersed in ether solution at room temperature carries out ungrease treatment,
Ensure that ether solution floods all skeletal grains completely；Treat after the completion of ungrease treatment with deionized water rinsing skeletal grain to remove ether；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 25 days~30 days；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out enzyme at
Reason；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo afterwards, then by dried skeletal grain
Sterilization treatment is carried out, that is, obtains bone grafting material.
The features of the present invention also resides in：
In step 2：The mass percent concentration of liquor natrii hypochloritis be 0.06%~0.08%, soak time be 12h~
15h。
In step 3：The time that deionized water is soaked is used as 26h~32h.
In step 4：The mass percent concentration of ether solution is 85%~90%.
In step 5：The condition of freezing processing is：The liquid nitrogen frozen under normal pressure, temperature control are -150 DEG C~-100 DEG C.
In step 6：The concentration of trypsin solution is 0.2g/L~0.3g/L, and soak time is 15h~18h.
In step 7：Sterilization treatment is irradiated using gamma-rays, and irradiation sterilization dosage is 20KGy~22KGy, irradiation time
For 5h~6h.
The beneficial effects of the present invention are：
(1) a kind of preparation method of bone grafting material of the present invention, can prepare homogeneous allogenic bone derived material, very suitable
The reparation for Cranial defect etc. is closed, is adapted to application clinically；
(2) a kind of preparation method of bone grafting material of the present invention, when being prepared different from existing homogeneous allogenic bone derived material
The mode drawn materials from people's bone tissue, the propagation for the disease that can keep off infection to a certain extent, and also materials will not also receive limit
System；
(3) a kind of preparation method of bone grafting material of the present invention, process is simple, is easy to produce in enormous quantities.
Embodiment
With reference to embodiment, the present invention is described in detail.
A kind of preparation method of bone grafting material of the present invention, specifically implements according to following steps：
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, rushed under aseptic condition with deionized water
To wash so that bone tissue does not show color, its object is to：Remove the marrow and haemocyte in ossis；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, are killed potential
Virus, finally rinse bone tissue surface repeatedly with deionized water；
Wherein, the mass percent concentration of liquor natrii hypochloritis is 0.06%~0.08%, and soak time is 12h~15h；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, will obtain
Skeletal grain using deionized water immersion 26h~32h, then to skeletal grain carry out centrifugal treating remove impurity therein；
Step 4, the skeletal grain after step 3 is handled is immersed in ether solution at room temperature carries out ungrease treatment, be
Make every skeletal grain can complete degreasing, it is ensured that ether solution floods all skeletal grains completely；Treat to use after the completion of ungrease treatment
Deionized water rinsing skeletal grain is to remove ether；
Wherein, the mass percent concentration of ether solution is 85%~90%；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 25 days~30 days, it is therefore intended that：Pass through the side of freezing
Formula destroys osteocyte, so as to reduce antigenicity；
Wherein, the condition of freezing processing is：The liquid nitrogen frozen under normal pressure, temperature control are -150 DEG C~-100 DEG C；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out enzyme at
Reason；
Wherein, the concentration of trypsin solution is 0.2g/L~0.3g/L, and soak time is 15h~18h；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo afterwards, then by dried skeletal grain
Sterilization treatment is carried out, that is, obtains the bone grafting material of the present invention；
Wherein, sterilization treatment is irradiated using gamma-rays, and irradiation sterilization dosage is 20KGy~22KGy, irradiation time 5h
~6h.
Embodiment 1
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, rushed under aseptic condition with deionized water
Wash so that bone tissue does not show color；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, are killed potential
Virus, finally rinse bone tissue surface repeatedly with deionized water；
Wherein, the mass percent concentration of liquor natrii hypochloritis is 0.06%, soak time 12h；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, will obtain
Skeletal grain using deionized water immersion 26h, then to skeletal grain carry out centrifugal treating remove impurity therein；
Step 4, the skeletal grain after step 3 is handled is immersed in ether solution at room temperature carries out ungrease treatment,
Ensure that ether solution floods all skeletal grains completely；Treat after the completion of ungrease treatment with deionized water rinsing skeletal grain to remove ether；
Wherein, the mass percent concentration of ether solution is 85%；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 25 days；
Wherein, the condition of freezing processing is：The liquid nitrogen frozen under normal pressure, temperature control are -150 DEG C；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out enzyme at
Reason；
Wherein, the concentration of trypsin solution is 0.2g/L, soak time 15h；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo afterwards, then by dried skeletal grain
Sterilization treatment is carried out, that is, obtains the bone grafting material of the present invention；
Wherein, sterilization treatment is irradiated using gamma-rays, and irradiation sterilization dosage is 20KGy, irradiation time 5h.
Embodiment 2
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, rushed under aseptic condition with deionized water
Wash so that bone tissue does not show color；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, are killed potential
Virus, finally rinse bone tissue surface repeatedly with deionized water；
Wherein, the mass percent concentration of liquor natrii hypochloritis is 0.065%, soak time 13h；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, will obtain
Skeletal grain using deionized water immersion 28h, then to skeletal grain carry out centrifugal treating remove impurity therein；
Step 4, the skeletal grain after step 3 is handled is immersed in ether solution at room temperature carries out ungrease treatment, be
Make every skeletal grain can complete degreasing, it is ensured that ether solution floods all skeletal grains completely；Treat to use after the completion of ungrease treatment
Deionized water rinsing skeletal grain is to remove ether；
Wherein, the mass percent concentration of ether solution is 86%；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 26 days；
Wherein, the condition of freezing processing is：The liquid nitrogen frozen under normal pressure, temperature control are -130 DEG C；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out enzyme at
Reason；
Wherein, the concentration of trypsin solution is 0.25g/L, soak time 16h；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo afterwards, then by dried skeletal grain
Sterilization treatment is carried out, that is, obtains the bone grafting material of the present invention；
Wherein sterilization treatment is irradiated using gamma-rays, and irradiation sterilization dosage is 21KGy, irradiation time 5.5h.
Embodiment 3
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, rushed under aseptic condition with deionized water
Wash so that bone tissue does not show color；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, are killed potential
Virus, finally rinse bone tissue surface repeatedly with deionized water；
Wherein, the mass percent concentration of liquor natrii hypochloritis is 0.07%, soak time 14h；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, will obtain
Skeletal grain using deionized water immersion 29h, then to skeletal grain carry out centrifugal treating remove impurity therein；
Step 4, the skeletal grain after step 3 is handled is immersed in ether solution at room temperature carries out ungrease treatment, be
Make every skeletal grain can complete degreasing, it is ensured that ether solution floods all skeletal grains completely；Treat to use after the completion of ungrease treatment
Deionized water rinsing skeletal grain is to remove ether；
Wherein, the mass percent concentration of ether solution is 87%；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 27 days；
Wherein, the condition of freezing processing is：The liquid nitrogen frozen under normal pressure, temperature control are -120 DEG C；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out enzyme at
Reason；
Wherein, the concentration of trypsin solution is 0.25g/L, soak time 17h；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo afterwards, then by dried skeletal grain
Sterilization treatment is carried out, that is, obtains the bone grafting material of the present invention；
Wherein sterilization treatment is irradiated using gamma-rays, and irradiation sterilization dosage is 20KGy, irradiation time 5.5h.
Embodiment 4
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, rushed under aseptic condition with deionized water
Wash so that bone tissue does not show color；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, are killed potential
Virus, finally rinse bone tissue surface repeatedly with deionized water；
Wherein, the mass percent concentration of liquor natrii hypochloritis is 0.075%, soak time 14.5h；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, will obtain
Skeletal grain using deionized water immersion 31h, then to skeletal grain carry out centrifugal treating remove impurity therein；
Step 4, the skeletal grain after step 3 is handled is immersed in ether solution at room temperature carries out ungrease treatment, be
Make every skeletal grain can complete degreasing, it is ensured that ether solution floods all skeletal grains completely；Treat to use after the completion of ungrease treatment
Deionized water rinsing skeletal grain is to remove ether；
Wherein, the mass percent concentration of ether solution is 89%；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 28 days；
Wherein, the condition of freezing processing is：The liquid nitrogen frozen under normal pressure, temperature control are -110 DEG C；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out enzyme at
Reason；
Wherein, the concentration of trypsin solution is 0.3g/L, soak time 17h；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo afterwards, then by dried skeletal grain
Sterilization treatment is carried out, that is, obtains the bone grafting material of the present invention；
Wherein sterilization treatment is irradiated using gamma-rays, and irradiation sterilization dosage is 21KGy, irradiation time 5h.
Embodiment 5
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, rushed under aseptic condition with deionized water
Wash so that bone tissue does not show color；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, are killed potential
Virus, finally rinse bone tissue surface repeatedly with deionized water；
Wherein, the mass percent concentration of liquor natrii hypochloritis is 0.08%, soak time 15h；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, will obtain
Skeletal grain using deionized water immersion 32h, then to skeletal grain carry out centrifugal treating remove impurity therein；
Step 4, the skeletal grain after step 3 is handled is immersed in ether solution at room temperature carries out ungrease treatment, be
Make every skeletal grain can complete degreasing, it is ensured that ether solution floods all skeletal grains completely；Treat to use after the completion of ungrease treatment
Deionized water rinsing skeletal grain is to remove ether；
Wherein, the mass percent concentration of ether solution is 90%；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 30 days；
Wherein, the condition of freezing processing is：The liquid nitrogen frozen under normal pressure, temperature control are -100 DEG C；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out enzyme at
Reason；
Wherein, the concentration of trypsin solution is 0.3g/L, soak time 18h；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo afterwards, then by dried skeletal grain
Sterilization treatment is carried out, that is, obtains the bone grafting material of the present invention；
Wherein sterilization treatment is irradiated using gamma-rays, and irradiation sterilization dosage is 21KGy, irradiation time 6h.
Claims (7)
1. a kind of preparation method of bone grafting material, it is characterised in that specifically implement according to following steps：
Step 1, by health ox bone, sheep bone or pig bone in weeding out soft tissue, deionized water rinsing is used under aseptic condition, is made
Obtain bone tissue and do not show color；
Step 2, the ox bone by after step 1 is handled, sheep bone or pig bone are soaked using liquor natrii hypochloritis, kill potential disease
Poison, finally rinse bone tissue surface repeatedly with deionized water；
Step 3, after step 2, by ox bone, sheep bone or pig bone shred, turn into the mutually uniform skeletal grain of particle size, the bone that will be obtained
Grain is soaked using deionized water, then is carried out centrifugal treating to skeletal grain and removed impurity therein；
Step 4, the skeletal grain after step 3 is handled be immersed in ether solution at room temperature carry out ungrease treatment, it is ensured that
Ether solution floods all skeletal grains completely；Treat after the completion of ungrease treatment with deionized water rinsing skeletal grain to remove ether；
Step 5, the skeletal grain obtained through step 4 is subjected to freezing processing 25 days~30 days；
Step 6, under the conditions of 35 DEG C, the skeletal grain obtained through step 5 is dipped into trypsin solution, carry out ferment treatment；
Step 7, the skeletal grain deionized water rinsing to being obtained through step 6, are dried in vacuo, then dried skeletal grain is carried out afterwards
Sterilization treatment, that is, obtain bone grafting material.
2. the preparation method of a kind of bone grafting material according to claim 1, it is characterised in that in the step 2：Secondary chlorine
The mass percent concentration of acid sodium solution is 0.06%~0.08%, and soak time is 12h~15h.
3. the preparation method of a kind of bone grafting material according to claim 1, it is characterised in that in the step 3：Using
The time of deionized water immersion is 26h~32h.
4. the preparation method of a kind of bone grafting material according to claim 1, it is characterised in that in the step 4：Ether
The mass percent concentration of the aqueous solution is 85%~90%.
5. the preparation method of a kind of bone grafting material according to claim 1, it is characterised in that in the step 5：Freezing
The condition of processing is：The liquid nitrogen frozen under normal pressure, temperature control are -150 DEG C~-100 DEG C.
6. the preparation method of a kind of bone grafting material according to claim 1, it is characterised in that in the step 6：Pancreas egg
The concentration of white enzyme solutions is 0.2g/L~0.3g/L, and soak time is 15h~18h.
7. the preparation method of a kind of bone grafting material according to claim 1, it is characterised in that in the step 7：Sterilizing
Processing is irradiated using gamma-rays, and irradiation sterilization dosage is 20KGy~22KGy, and irradiation time is 5h~6h.
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| CN105169480A (en) * | 2015-09-06 | 2015-12-23 | 中国人民解放军总医院第一附属医院 | Scaffold material applied to vertebral fusion |
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2017
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| EP2783707A1 (en) * | 2011-11-23 | 2014-10-01 | Chungbuk National University Industry-Academic Cooperation Foundation | Production method for biomedical and industrial material using ceramic derived from birds' beaks |
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Application publication date: 20180116 |