CN103656762A - Application of silk fibroin film as absorbable tissue regeneration guiding film - Google Patents
Application of silk fibroin film as absorbable tissue regeneration guiding film Download PDFInfo
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- CN103656762A CN103656762A CN201210360333.6A CN201210360333A CN103656762A CN 103656762 A CN103656762 A CN 103656762A CN 201210360333 A CN201210360333 A CN 201210360333A CN 103656762 A CN103656762 A CN 103656762A
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Abstract
The invention belongs to the technical field of medical apparatus and instruments and particularly relates to an application of a silk fibroin film as an absorbable tissue regeneration guiding film. The silk fibroin film applied as the absorbable tissue regeneration guiding film can be used for preparing dental implants and treating periodontal diseases. The silk fibroin film is used as the absorbable tissue regeneration guiding film in the dental department and is single in raw material source, low in production cost, nontoxic and free of environment pollution.
Description
Technical field
The invention belongs to technical field of medical instruments, specifically, relate to fibroin protein film as the application of absorbability tissue regeneration guiding film.
Background technology
Film inductive bone regeneration is a new biological concept; its ultimate principle is film to be placed in to soft tissue and bone is set up biological barrier between damaged; stop to disturb bone formation and migration rate gum connective tissue cell and epithelial cell enters and allow potential regeneration capacity, the slower osteocyte of migration rate preferentially to enter bone defective region faster; dominant growth; protect blood clot simultaneously; slow down the pressure that covers tissue, realize the Regeneration and Repair of defective region osseous tissue.
Film guide regeneration technology (MGTR) is not applied in periodontal disease therapeutic at first, MGTR technology is applied to after the treatment success of periodontal disease, phase late 1980s Dahlin etc. introduces oral implantology by MGTR, what in MGTR technology, play a crucial role is guiding film, the absorbability film that the collagen film of wherein take is representative is due to the absorbability of himself, without second operation, take out, greatly alleviated patient misery and extremely Chinese scholars praise highly.
Collagen film is current domestic and international application natural biological medical material the most widely.But collagen film has a lot of irremediable defects: the material source of collagen film mixes, and standardized production difficulty is larger; In the process of collagen protein masking from cattle and Os Caprae seu Ovis purification, must add cross-linking agent that tropocollagen molecule is linked together, technique is loaded down with trivial details; Production cost is large.
Scientist is striving to find a kind of desirable new type natural bio-medical material replacement collagen film always both at home and abroad.
Fibroin albumen is the natural polymer fibrin extracting from silkworm silk, and content accounts for the 70-80% of silkworm silk, has 18 seed amino acids.Because fibroin albumen has the compatibility of good biological tissue, biology, chemistry and physics characteristic, can make up the deficiency of collagen protein, and meet clinical requirement completely, be rare biological medicine engineering material.Fibroin albumen is studied widely in field of biomedical materials at present.Experiment showed, that fibroin protein film not only can replace collagen film completely as a kind of novel natural polymer bio-medical material, and the lot of advantages of fibroin albumen is that collagen protein is completely incomparable.
Summary of the invention
Loaded down with trivial details in order to overcome the preparation technology that collagen film exists as dentistry absorbability regeneration guiding film, high in cost of production drawback, what the invention provides that a kind of cost is low, raw material sources are single can be applicable to dentistry absorbability tissue regeneration guiding film.
The invention provides fibroin protein film as the application of absorbability tissue regeneration guiding film.
In the present invention, the preparation method of fibroin protein film referring to Chinese patent application (application number: 200910060497, denomination of invention: a kind of tissue regeneration membrane with bioactivity and preparation method thereof).
The invention provides fibroin protein film as the application of absorbability tissue regeneration guiding film, wherein absorbability regeneration guiding film is prepared for Dental Implant.
Dental Implant is prepared: for the preoperative trimestral alveolar bone of Dental Implant hole, repair.After loss of tooth, on alveolar bone, leave over a dark hole, must, through processing, by the bone that makes new advances of growing in tooth bone hole, can on alveolar bone, plant artificial tooth.
This processing procedure comprises three steps: first in alveolar bone hole, place bone meal or bone substitute; Then on the surface of populated bone meal, with adsorbable bone tissue regeneration guiding film, cover completely; Finally gingiva tissue is wrapped up.
The invention provides fibroin protein film as the application of absorbability tissue regeneration guiding film, wherein absorbability regeneration guiding film is used for the treatment of periodontal disease.
Periodontal disease is a kind of whole mankind's frequently-occurring disease, in the past except removing tartar without any effective Therapeutic Method.The early 1990s is brought into use guided bone paradenlal tissue regeneration art treatment periodontal disease.Because therapeutic effect is remarkable, a lot of patients can cure, so guided bone paradenlal tissue regeneration art treatment periodontal disease is more and more extensively by doctor and the clinical acceptance of patient, popularization both at home and abroad.Guided bone paradenlal tissue regeneration art is to utilize film material as barrier, stop gingival epithelium in agglutination along the root length of looking unfamiliar, stop contacting of gingival connective tissue and root face, and provide certain space, the periodontal ligament cell that guiding has the new adhesive ability of formation preferentially captures root face, thereby on the former root face being exposed in periodontal pocket, form new cementum, and have periodontal membrane fiber to imbed, form new tack healing.
Beneficial effect of the present invention is:
Fibroin protein film is pure-natural biological material, has good pliability and tensile strength, has utmost point low antigenicity, good hydrophilic property, has controlled degradation speed in vivo and in vitro, is applied in dentistry, as absorbability regeneration guiding film, its raw material source is single, and production cost is low; Nontoxic, non-environmental-pollution.
Accompanying drawing explanation
Fig. 1, Fig. 1 a is cell incremental analysis figure; Fig. 1 b is that cell is cultivated the Electronic Speculum figure after 3 days on fibroin protein film.
Fig. 2, external degradation analysis chart
.
Fig. 3, biocompatibility analysis result in body.
The specific embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail.
Embodiment 1
In film guide regeneration technology, the biomembrane of desirable dental applications need to possess the features such as the compatibility of good biological tissue, degradability, reliable mechanical performance, hydrophilic.For biology, chemistry and physics characteristic from theory and practice checking fibroin protein film, the present invention has done following experiment, the patent of invention embodiment 1 that wherein preparation method of fibroin protein film is 200910060497 with application number.
1) in vitro tests
A. cell in vitro adsorption test: experimental technique adopts contact method cytotoxicity experiment referring to (" preparation of collagen-gelatin timbering material modification and cytotoxicity experiment research ", biomedical engineering and clinical 2007, V11,6).
Cell adsorption experiment concrete steps are as follows:
(1) preparation of cell suspending liquid and culture dish.1. suspension preparation: with the DMEM/F12 culture fluid cultured cell L929 containing 10 % hyclones, make the cell suspending liquid of 1 * 105/ml in 150 ml Tissue Culture Flasks.2. culture dish preparation: add 5～7 of 2 mlDMEM/F12 culture fluid and cell suspending liquids in culture dish, hatch 3～5 d, until it is fused into approximate cell monolayer.
(2) contact method toxicity test method and grading standard.Observation of cell growing state under inverted phase contrast microscope, makes fibroin protein film directly contact with cell monolayer, and after 24 h are cultivated in continuation, observation of cell metamorphosis is also taken a picture.
In this experiment, adopted osteoblast, mescenchymal stem cell, periodontal cell and gingival cell to cultivate.Then observe the absorption number of various cells on fibroin protein film.
As shown in Figure 1, result shows result of the test: along with the prolongation of time of cell culture, the cell number of the absorption of various cells on fibroin protein film increases gradually.These various cells that show fibroin protein film and human body have fine biocompatibility, without any rejection.
B. external fibroin protein film slow degradation experiment: biodegrading process is with reference to (" the biodegradation Journal of Sex Research of natural silk and silk fibroin porous film ", chemical journal, 2007, V65,22).
Degrading experiment is specific as follows: trypsin in experiment, protease is dissolved in to pH=7, concentration is in the phosphate buffered solution of 0.05mol/L, to obtain degraded trypsin enzymatic solution, at official hour point, take out the fibroin albumen membrane sample being placed in one, change enzymatic solution, to guarantee the vigor of enzymatic solution simultaneously.In experiment, prepare a duplicate samples and be placed in not enzyme-added buffer solution and compare, and according to the percentage loss of weight of following formula calculation sample.
Wherein
wfor example weight after degrading,
w 0for the front example weight of degrading.
As shown in Figure 2, under the condition existing at protease, experiment in vitro is in the time of 42 days for degradation experiment result, and more than 40% fibroin albumen is degradable under the existence of enzyme, and breaks down into amino acids is degradable in the time of 100 days greatly.
C. mechanical robustness testing:
Test method referring to (Standard Test for Plane Strain Fracture Toughness of Metallic Materials,
ASTME-399，American?society?for?testing?and?materials,?Philadelphia,?PA,?1987）
Result shows that fibroin protein film has good tensile characteristics and toughness, it is under the test condition without buffer, modulus is (3.24 ± 1.98) MPa, and surrender tensile strength is (0.64 ± 0.12) MPa, and the maximum tensile strength is (1.59 ± 0.87) MPa; Under the condition of 37 ℃ of buffer, test, its modulus is (4.98 ± 1.98) MPa, and surrender tensile strength is (1.01 ± 0.12) MPa, and the maximum tensile strength is (1.66 ± 0.87) MPa.
D. hydrophilic test: testing method is with reference to " test philosophy of common hardness and method for expressing; metal processing (measure and control); 2008; (15): 69-71 ", result shows that the contact angle of fibroin protein film in Methanamide is 62 degree, contact angle in glycerol is 73 degree, and this shows that fibroin protein film has good hydrophilic.
2) in vivo test
Fibroin protein film is implanted in the lower limb muscles of mice, then two weeks, surrounding and each sampling in eight weeks, the tectology of examining under a microscope implant site changes.Be illustrated in figure 3 biocompatibility analysis result in body, fibroin protein film has the compatibility of good biological tissue, slowly degraded.Without immunoreation and inflammatory reaction.
No matter from bibliographical information or our result of the test can prove that fibroin protein film is a kind of desirable natural biological medical material, can replace collagen protein completely, as absorbability regeneration guiding film, be applied to periodontal disease therapeutic and Dental Implant is prepared.
Claims (3)
1. fibroin protein film is as the application of absorbability tissue regeneration guiding film.
2. application according to claim 1, is characterized in that: described absorbability tissue regeneration guiding film is prepared for Dental Implant.
3. application according to claim 1, is characterized in that: described absorbability tissue regeneration guiding film is used for the treatment of periodontal disease.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210360333.6A CN103656762A (en) | 2012-09-25 | 2012-09-25 | Application of silk fibroin film as absorbable tissue regeneration guiding film |
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| CN201210360333.6A CN103656762A (en) | 2012-09-25 | 2012-09-25 | Application of silk fibroin film as absorbable tissue regeneration guiding film |
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| CN103656762A true CN103656762A (en) | 2014-03-26 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106535950A (en) * | 2014-08-04 | 2017-03-22 | 大韩民国（农村振兴厅长） | Dental barrier membrane using cocoon and method for manufacturing same |
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2012
- 2012-09-25 CN CN201210360333.6A patent/CN103656762A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106535950A (en) * | 2014-08-04 | 2017-03-22 | 大韩民国（农村振兴厅长） | Dental barrier membrane using cocoon and method for manufacturing same |
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Application publication date: 20140326 |