CN1739809A - A kind of silk fibroin coated titanium mesh and its preparation method and application - Google Patents
A kind of silk fibroin coated titanium mesh and its preparation method and application Download PDFInfo
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Abstract
本发明属组织工程和医用材料领域，具体涉及一种丝素蛋白涂层钛网及其制备方法和在气管缺损重建中应用。本发明采用医用钛网作为支撑材料，以丝素蛋白为涂层，制成丝素蛋白涂层钛网复合体。本发明利用丝素的良好生物相容性和促进成纤维细胞和粘膜细胞生长的作用，能加速愈合，减少并发症，又可以长期或永久有效支撑管腔。本发明丝素蛋白涂层的钛网可用作临床气管缺损修补，能为探讨丝素蛋白在促进气管粘膜修复及临床重建气管缺损、减少重建气管术后并发症的作用提供有意义的参考。The invention belongs to the field of tissue engineering and medical materials, and in particular relates to a silk fibroin-coated titanium mesh, a preparation method thereof and an application in tracheal defect reconstruction. The invention adopts medical titanium mesh as supporting material and silk fibroin as coating to make a silk fibroin-coated titanium mesh complex. The invention utilizes the good biocompatibility of silk fibroin and the effect of promoting the growth of fibroblasts and mucosal cells, can accelerate healing, reduce complications, and can effectively support the lumen for a long time or permanently. The titanium mesh coated with silk fibroin of the present invention can be used as clinical tracheal defect repair, and can provide meaningful reference for exploring the role of silk fibroin in promoting tracheal mucosa repair and clinical reconstruction of tracheal defect, and reducing complications after tracheal reconstruction.
Description
Technical field
The invention belongs to organizational project and field of medical materials, be specifically related to a kind of titanium net with fimbrin and preparation method thereof and application in trachea defect is rebuild.
Background technology
At present, body is causing because of inflammation, wound, tumor or other pathological changes that trachea is impaired on a large scale, under the situation of the long section trachea of excision, is needing the repair deficiency trachea.For a long time, the reparation problem that tracheal cartilages is damaged is perplexing the clinician always.The first phase end-to-end anastomosis only is suitable for the reparation of SOL, when defect length surpasses 6.0cm, easily produces severe complications such as anastomotic leakage because anastomotic stoma tension force is high.Prominent the flying of tissue engineering technique research brought new hope to tracheal reconstruction suddenly, but do not have breakthrough from 1994 again after preliminary the trial, still is in experiment and exploratory stage, still has problem to need to be resolved hurrily.Many scholars attempt using methods such as allograft, autograft and artificial material substitute, and effect is all not ideal enough.It is how to introduce at the artificial trachea surface of internal cavity to have the material of regulating the effect of tunica mucosa tracheae cell growth metabolism that artificial trachea improve to be used more crucial, promotes the mucous epithelium regeneration of creeping.In numerous biomaterials, collagen is considered to comparatively ideal selection.Yet collagen belongs to allosome animal tissue extract, has the potential probability of diseases such as virus spread, and complex manufacturing process, and fibroin belongs to native protein, and the source is abundant, and price is suitable.The foreign scholar has studied the different growing state of human body cell strain on fibroin protein film, and the result shows that fibroin is the same with collagen protein, can promote the growth of cell.
Fibroin albumen is a kind of natural structure albumen, and mainly by three kinds of simple aminoacid: glycine, alanine and serine are formed, and they account for 85% of Tot Prot.Fibroin membrane is good at the biocompatibility at positions such as skin, oral cavity and eyes, and antigenicity is not obvious, and the subdermal implantation test shows that no obvious inflammation or occupy-place phenomenon take place.There are some researches show: the amino acid residue owing to band polar groups such as the OH, the COOH that not only contain significant proportion in the fibroin albumen, NH2 makes the hydrophilic of regenerated silk fibroin film strong; Fibroblast is because of its film surface fibre rich Fibronectin, histone etc., and strong with the ability of biomaterial surface adhesive attraction, the structural information that regenerated silk film surface is entrained obviously has important function to fibroblastic adhesion and propagation.Also contain a certain proportion of positively charged amino acid residue on the other hand, this has also created good condition for fibroblast in the adhesion and the propagation on regenerated silk film surface.
Fibroin albumen has excellent biological compatibility, and is nontoxic, and nonirritant has obtained increasingly extensive application in the bio-medical field, as experimentation of medical fibroin protein skin regeneration film, contact lens, cell culture medium and artificial meninges or the like.But up to now, still there is not the report that fibroin albumen is used both at home and abroad aspect the trachea defect reconstruction.
Summary of the invention
The purpose of this invention is to provide a kind of titanium net with fimbrin.Another object of the present invention provides the method for the described titanium net of preparation.Further purpose of the present invention provides the application of described titanium net in clinical trachea defect is rebuild.
The present invention adopts the medical titanium net as backing material, is coating with the fibroin albumen, makes the titanium net with fimbrin complex, and described fibroin albumen concentration is 6-12% (weight/volume), and described titanium mesh crack is not sealed.The present invention utilizes the good biocompatibility of fibroin and promotes fibroblast and the effect of mucomembranous cell growth, can healing acceleration, reduce complication, and again can be for a long time or permanent effectively support tube chamber.
The titanium net of fimbrin of the present invention can be used as clinical trachea defect to be repaired, and can provide significant reference in the effect that promotes tunica mucosa tracheae reparation and clinical reconstruction trachea defect, minimizing to rebuild the trachea post-operative complication for inquiring into fibroin albumen.
Titanium net with fimbrin of the present invention prepares by following method and step:
1, preparation fibroin albumen
Adopt useless Bombyxmori Linnaeus raw silk in NaHCO3 (0.5wt%) aqueous solution, to come unstuck, degumed silk is dissolved in the LiBr aqueous solution of 9.3mol/L, be stirred to silkworm silk and dissolve fully.Filter, remove undissolved silkworm silk and impurity, the fibroin albumen saline solution, remove LiBr in the solution with the deionized water dialysis, concentration is 4% fibroin, reconcentration, the PEG dialysis, the centrifugal removal impurity of gained silk fibroin protein solution stays supernatant.The fibroin albumen concentration of gained is 20%, puts 4 ℃ of refrigerators and preserves standby.By the concentration of weight method mensuration silk fibroin protein solution, can dilute solution as required for the 6-12% variable concentrations, avoid causing the variation of conformation simultaneously.
2, preparation fibroin coated titanium net
Adopt medical titanium net (commercial) to make and the shape with certain elastic class annulus trachealis spline structure laboratory animal trachea identical radian measure, that thickness is identical, after the epoxyethane fumigation sterilization, it is the silk fibroin solution of 6-12% that the titanium net is dipped the diluted concentration of preserving in above-mentioned low temperature (4 degrees centigrade) repeatedly, put into baking oven 60 degree oven dry after the taking-up, 6-10 time so repeatedly, make its surface evenly be coated with silk fibroin material, and titanium mesh crack is not sealed, the equal sterile working of above process.
3, trachea defect is rebuild zoopery
Cleaning level 25 of male White Rabbits of New Zealand (providing by Fudan University zoopery center) are provided, body weight 2.3-2.5KG, average 2.4KG, rabbit is 3 months age, is divided at random: the titanium net group (n=12) of fimbrin, simple titanium net group (n=12) are arranged.Normal rabbits (n=1) in contrast.
The result:
1) the titanium net reconstruction group of fibroin coating is arranged:
The equal apnea difficulty of all rabbit postoperatives, 4 weeks of postoperative, 8 weeks, 12 weeks draw materials respectively, all see tracheal reconstruction place structure ideal, and fibroin titanium net surface covers growth translucent fibre tissue, and has the blood capillary growth to see in the tissue, and does not have adhesion with surrounding tissue.
The trachea splanchnoscopy is seen trachea intracavity smooth mucosal, and tube chamber does not have obvious stenosis, no cicatrix and granulation tissue growth.
Trachea CT examination or cut the capable CT examination of reconstruction place trachea there is no obvious tracheal stenosis, and CT only has the extremely slight localized thickening of part tunica mucosa tracheae, and AIR STRUCTURE iconography is rebuild, and reconstruction place trachea chamber also shows no narrow.
The tissue pathological slice inspection sees that damaged reconstruction place mucomembranous cell is formed and normal tunica mucosa tracheae no significant difference, mainly contains compositions such as ciliated columnar and goblet cell.
Scanning electron microscope checks that well-grown during 8 weeks, cilium are long, closeer, but directivity is still poor slightly and than normal tunica mucosa tracheae cilium weak point and sparse, recover normal during 12 weeks, with normal tunica mucosa tracheae cilium no significant difference.
2) simple titanium net reconstruction group
All rabbit postoperatives are in various degree concurrent cervical region subcutaneous emphysema all, 2 routine concurrent whole body subcutaneous emphysemas wherein, and diarrhoea appearred in 14 days in 1 routine rabbit behind heeling-in postoperative 5 days and Reconstruction, other one routine rabbit 17 days the time, diarrhoea occurs behind Reconstruction, do not take food, become thin death gradually.Other has the subcutaneous lump of the concurrent cervical region of rabbit postoperative, sees tracheal reconstruction place titanium net surface scar tissue when drawing materials.
Dead rabbit trachea splanchnoscopy sees that the trachea intracavity infects sticking purulent core formation and the red inflammatory granulation tissue growth of reconstruction place, and the tube chamber obvious stenosis has the growth of cicatrix and granulation tissue, and tangible secretions is arranged.
The trachea CT examination is seen trachea chamber severe stenosis, even does not have lacuna fully,
Histopathological examination shows a large amount of granulation tissue hyperplasias, and a small amount of inflammatory cell infiltration, and the fibrous tissue layer is than fibrous tissue bed thickness under the titanium net reconstruction group mucosa of fibroin coating under the mucosa, and cilium is more sparse.
Scanning electron microscope inspection demonstration compares with the titanium net reconstruction group of fibroin coating that cilium is more sparse, short, directivity is poor, self-purification function is poor, and the surface has a little secretions to adhere to
3) use the SAS6.12 statistical software, with cmh card side, the ridit scoring of methods of marking for revising, and check the difference of the interior area of section of the difference of cross-sectional area of two groups of hyperplastic tissues that rebuild the stenosis of tracheas and corresponding section trachea with the time factor layering; Check two groups of other post-operative complication (deaths by suffocation with definite probabilistic method; Subcutaneous emphysema; Cervical infection) difference.Table 1 is post-operative complication " death by suffocation " contrast.Table 2 is the concurrent cervical infection situations of postoperative.Table 3 is subcutaneous emphysema situations.
Table 1
The death by suffocation total that do not occur suffocating
The titanium net reconstruction of fibroin coating organizes 0 11 11
Simple titanium net reconstruction organizes 56 11
Add up to 5 17 22
Definite probabilistic method check: significant difference has significance (p＜0.05).
Table 2
The cervical infection cervical region does not have the total of infection
The titanium net reconstruction of fibroin coating organizes 0 11 11
Simple titanium net reconstruction organizes 29 11
Add up to 2 20 22
Definite probabilistic method check: significant difference does not have significance (p＞0.05).
Table 3
Subcutaneous emphysema occurs and do not have the subcutaneous emphysema total
The titanium net reconstruction of fibroin coating organizes 38 11
Simple titanium net reconstruction organizes 11 0 11
Add up to 14 8 22
Definite probabilistic method check: significant difference has significance (p＜0.05=.
4) use 16 row's spiral CT (U.S. GE company products) AW4.0 work station to carry out graphical analysis, calculate the cross-sectional area of hyperplastic tissue of the stenosis of rabbit reconstruction trachea and the interior area of section of corresponding section trachea.Table 4 is cross-sectional areas (mm2) that rabbit is rebuild the hyperplastic tissue of the stenosis of trachea.Table 5 is interior areas of section of corresponding section trachea.
Table 4
| The rabbit of suffocating when rebuilding for 4 weeks and in 4 weeks | When rebuilding for 8 weeks | When rebuilding for 12 weeks | |
| Fibroin coated titanium net reconstruction group | 4.02， 1.35， 1.41 | 4.05， 5.31， 3.35， 5.78 | 3.56， 2.29， 2.25， 1.24 |
| Simple titanium net reconstruction group | 28.56，21.75，26.31， 25.76，12.78，16.65 | 14.80，24.89， 9.21 | 14.56，4.35 |
The result shows, the titanium net reconstruction group rabbit of fibroin coating is rebuild the cross-sectional area of the hyperplastic tissue of the stenosis of trachea: minima is 1.24mm
2, maximum is 5.78mm
2, average out to 3.15 ± 1.58mm
2, median is 3.35.
Simple titanium net reconstruction group rabbit is rebuild the cross-sectional area of the hyperplastic tissue of the stenosis of trachea: minima is 4.35mm
2, maximum is 28.56mm
2, average out to 18.15 ± 7.85mm
2, median is 16.65.
Two groups of data: with cmh card side, the ridit scoring of methods of marking for revising, and check the difference of the cross-sectional area of two groups of hyperplastic tissues that rebuild the stenosis of tracheas with the time factor layering, significant difference has significance (p＜0.05).
Table 5:
| The rabbit of suffocating when rebuilding for 4 weeks and in 4 weeks | When rebuilding for 8 weeks | When rebuilding for 12 weeks | |
| Fibroin coated titanium net reconstruction group | 4.02， 1.35， 1.41 | 4.05， 5.31， 3.35， 5.78 | 3.56， 2.29， 2.25， 1.24 |
| Simple titanium net reconstruction group | 28.56，21.75，26.31， 25.76，12.78，16.65 | 14.80，24.89， 9.21 | 14.56，4.35 |
The result shows that the interior sectional area at the titanium net reconstruction group rabbit tracheal reconstruction place of fibroin coating: minima is 26.79mm2, and maximum is 29.69mm2, average out to 28.37 ± 0.86mm2.
The interior sectional area at simple titanium net reconstruction group rabbit tracheal reconstruction place: minima is 26.31mm2, and maximum is 29.46mm2, average out to 28.13 ± 1.01mm2.
Two groups of data: check two groups of differences of rebuilding the interior area of section of trachea place trachea with the time factor layering, significant difference does not have significance (p=0.19＞0.05), and, the check of time layering factor, significant difference does not also have significance (p=0.35＞0.05).
The present invention shows through the zoopery result, fibroin can obviously promote growth of fibroblasts, the fibroblast close attachment is on the fibroin surface, the direction unanimity, and be polar growth is arranged, experimental result of the present invention shows that also blood capillary and trachea mucomembranous cell induce the fibroblast superficial growth of generation good at fibroin, no granulation tissue hyperplasia.The present invention finds that the epontic fibroblast of fibroin is not unrestricted propagation, but has certain polarity, level and thickness, but different fully at the fibroin surface thickness of different parts and different area.
The present invention adopts the medical titanium net as backing material, both can utilize the good biocompatibility of fibroin and promote fibroblast and the effect of mucomembranous cell growth, and healing acceleration reduces complication, again can be for a long time or permanent effectively support tube chamber.Compare with simple titanium net, the tracheal reconstruction that the titanium net that has a fimbrin is used for rabbit has more advantage, because fibroin can promote the growth of fibroblast and blood capillary, the complete film of the translucent fibrous tissue of titanium net surface before the trachea after two weeks of heeling-in closes all mesh, be used for tracheal reconstruction this moment, can the complete closed tracheal wall, can not communicate inside and outside making it, avoided the generation of subcutaneous emphysema and local infection.The mucosa of the normal trachea in trachea defect edge is just along the thin layer fibroblast superficial growth of fibroin coated titanium net surface, in time cover support tube chamber face apace fully, prevented granulation growth, tracheal stenosis and because the narrow generation that causes complication such as death by suffocation effectively.The Microscopic examination showed growth covers the mucosa well-grown of material surface, electron microscopic examination shows its cilium good directionality, self-purification function is as good as with normal tunica mucosa tracheae, and the titanium net can keep certain elasticity, length and enough hardness with the antagonism negative pressure as support, the titanium net of fimbrin is used for tracheal reconstruction does not find titanium net displacement phenomenon, the growth good knitting of the fibroblast of material surface and body residual gas tube edges tissue.
The present invention adopts the titanium net of fimbrin to be used for tracheal reconstruction, solved at the artificial trachea surface of internal cavity and introduced problem with material of regulating the effect of tunica mucosa tracheae cell growth metabolism, can promote mucous epithelium to creep, avoid the postoperative trachea narrow once again, obtained satisfied effect, had five standards of desirable tracheal replacement thing: be synthetic material with high degree of biocompatibility; Blood vessel suits to grow in graft; Tool epithelial tissue or be suitable for epithelial tissue growth; Sufficient length is arranged to satisfy defect repair in various degree; Has enough hardness with the antagonism negative pressure.Can further be generalized to clinical practice in the belt reconstruction damaged than large tracts of land tracheal tissue.
Description of drawings
The result detects in Fig. 1-3 endoscope,
A wherein: fibroin coated titanium net rebuild for 12 weeks, b: simple titanium net is rebuild the back granulation hyperplasia, c: simple titanium net is narrow when rebuilding for 12 weeks.
Fig. 2 is a fibroin coated titanium net reconstruction place mucomembranous cilium,
A wherein: 4 all reconstructions place of * 2000 postoperatives, b: 8 all reconstructions place of * 2000 postoperatives, c: 12 all reconstructions place of * 2000 postoperatives, recovered normal.
Fig. 3 is simple titanium net reconstruction place mucomembranous cilium,
A wherein: 4 all reconstructions place of * 2000 postoperatives, b: 8 all reconstructions place of * 2000 postoperatives, c: * 2000 postoperatives 12 all reconstructions place.
Fig. 4 is that fibroin coated titanium net is rebuild the CT examination result,
A wherein: rebuild for 4 weeks, b: rebuild for 8 weeks, c: rebuild for 12 weeks, d: sagittal plain is rebuild, and e:AIR STRUCTURE shows reconstruction place trachea chamber narrow (arrow refers to).
Fig. 5 is that simple titanium net is rebuild the CT examination result,
A wherein: rebuild for 8 weeks and suffocate b: it is narrow to rebuild for 12 weeks, and c: crown position rebuilds d: sagittal plain is rebuild, and e:AIR STRUCTURE shows reconstruction place trachea chamber narrow (arrow refers to).
The specific embodiment
Embodiment 1 preparation fibroin albumen
Adopt useless Bombyxmori Linnaeus raw silk in 100 ℃ NaHCO3 (0.5wt%) aqueous solution, to come unstuck 45 minutes, and degumed silk be dissolved in the LiBr aqueous solution of 9.3mol/L, 45 ℃ down continuous stir abouts 2 hours to silkworm silk dissolve fully.Filter with eight layers of hospital gauze, remove undissolved silkworm silk and impurity, the fibroin albumen saline solution that obtains is dialysed four days to remove the LiBr in the solution with deionized water, and the molecular weight that dams of bag filter is 12,000～14,000Da.Prepared fibroin concentration is about 4%, further it is concentrated, and the PEG that continues with 10% dialysed one day, and the silk fibroin protein solution that dialysis is good was removed impurity in centrifugal 15 minutes with the 6000rpm rotating speed, stayed the supernatant.The fibroin albumen concentration of gained is 20%, puts 4 ℃ of refrigerators as storage liquid and preserves standby.By the concentration of weight method mensuration silk fibroin protein solution, can dilute solution as required for the 6-12% variable concentrations.The sample that dialysis is good will avoid strong agitation with concussion and contact organic solvent etc. as far as possible, in case cause the variation of conformation, also will avoid stirring too violent in the dilution as far as possible.
Adopt medical titanium net (commercial) to make with laboratory animal rabbit trachea identical radian measure, the shape that thickness is identical with certain elastic class annulus trachealis spline structure, after the epoxyethane fumigation sterilization, it is 6% silk fibroin solution that the titanium net is dipped repeatedly in the diluted concentration of above-mentioned cryopreservation, put into baking oven 60 degree oven dry after the taking-up, 6-10 time so repeatedly, make its surface evenly be coated with silk fibroin material, and keep titanium mesh crack not seal the equal sterile working of above process.
Embodiment 3
Adopt medical titanium net (commercial) to make with laboratory animal rabbit trachea identical radian measure, the shape that thickness is identical with certain elastic class annulus trachealis spline structure, after the epoxyethane fumigation sterilization, it is 12% silk fibroin solution that the titanium net is dipped repeatedly in the diluted concentration of above-mentioned cryopreservation, put into baking oven 60 degree oven dry after the taking-up, 6-8 time so repeatedly, make its surface evenly be coated with silk fibroin material, and keep titanium mesh crack not seal the equal sterile working of above process.
Embodiment 4
Adopt medical titanium net (commercial) to make with laboratory animal rabbit trachea identical radian measure, the shape that thickness is identical with certain elastic class annulus trachealis spline structure, after the epoxyethane fumigation sterilization, it is 10% silk fibroin solution that the titanium net is dipped repeatedly in the diluted concentration of above-mentioned cryopreservation, put into baking oven 60 degree oven dry after the taking-up, 8 times so repeatedly, make its surface evenly be coated with silk fibroin material, and keep titanium mesh crack not seal the equal sterile working of above process.
Embodiment 5 zooperies
25 cleaning level male White Rabbits of New Zealand (providing) by Fudan University zoopery center, body weight 2.3-2.5KG, average 2.4KG, rabbit is 3 months age, is divided at random: the titanium net group (n=12) of fimbrin, simple titanium net group (n=12).Normal rabbits (n=1) in contrast.
1) the titanium net reconstruction group of fibroin coating:
Preset the heeling-in postoperative 7 days and Reconstruction after 12 days, every day, twice intramuscular injection penicillin was 100,000 u/ time.Postoperative was seldom permitted feed in three days, food-intake was normal substantially in the 5th day, postoperative body temperature is normal, substantially about 39.3-39.5 ℃, slight cervical region subcutaneous emphysema appears in 3 routine postoperatives, this organizes whole rabbit postoperatives does not have and infects complication and occur, and 14 days the time, diarrhoea occurs behind the routine rabbit Reconstruction, give gentamycin 10,000 units intramuscular injection/sky, diarrhoea stops after 3 days, but does not take food, and the back of becoming thin gradually is dead, the equal apnea difficulty of all rabbit postoperatives, postoperative has the slight sound of stridulating in 1 week, fade away 4 weeks of postoperative after the week, 8 weeks, 12 weeks drew materials respectively, all visible tracheal reconstruction place structure ideal, fibroin titanium net surface covers growth translucent fibre tissue, and has blood capillary to grow in the very thin tissue of this layer to see, and does not have adhesion with surrounding tissue.
The trachea splanchnoscopy:
Check when 4 weeks of postoperative, 8 weeks, 12 weeks, visible trachea intracavity smooth mucosal, tube chamber does not have obvious stenosis, no cicatrix and granulation tissue growth, mucomembranous surface has a little secretions when 4 weeks.
Family's rabbit anesthesia is fixed back trachea CT examination or is cut the capable CT examination of reconstruction place trachea:
All 4 weeks of rabbit postoperative, 8 weeks, 12 all trachea CT examination there is no obvious tracheal stenosis, and CT only has the extremely slight localized thickening of part tunica mucosa tracheae, and AIR STRUCTURE iconography is rebuild, and reconstruction place trachea chamber also shows no narrow.
The visible damaged reconstruction place mucomembranous cell of tissue pathological slice inspection is formed and normal tunica mucosa tracheae no significant difference, mainly contain compositions such as ciliated columnar and goblet cell, mucomembranous surface cilium growing state: postoperative was grown better during 4 weeks, subregion density is sparse slightly, cover well during 8 weeks, normal substantially during 12 weeks.Thin layer fibroblast ordering growth under the mucosa, the visible silk fibroin material good biocompatibility that does not absorb fully becomes fibrous tissue growth around the fibroin, accidental a little mononuclear cell, antigen-reactive is not obvious or do not have an obvious foreign body rejection.Compare with normal trachea pathological section (HE * 400), many one decks thin layer fibrous tissue under the former mucosa, and blood capillary is abundant.
Scanning electron microscope is checked: postoperative during 4 weeks cilium sparse, shorter, directivity is poor, self-purification function is poor, the surface has more secretions to adhere to, the subregion exposes fibre-less; Well-grown during 8 weeks, cilium are long, closeer, but directivity is still poor slightly and than normal tunica mucosa tracheae cilium weak point and sparse, self-purification function is better, the no obvious secretions in surface; Recover normal during 12 weeks, with normal tunica mucosa tracheae cilium no significant difference.
2) simple titanium net reconstruction group
Preset the heeling-in postoperative 7 days and Reconstruction after 12 days, every day, twice intramuscular injection penicillin was 100,000 u/ time, postoperative began a little feed on the 3rd day, food-intake was normal substantially in the 7th day, postoperative has behind the 2nd day to 4 days ears of 2 routine rabbit postoperatives about 40.5 ℃ of body temperature, wherein 1 routine postoperative is the 4th day, sticking purulent core forms and reconstruction place granulation causes death by suffocation jointly because infect in the trachea, other 1 example, the subcutaneous infection of postoperative one all throat portions forms fistula, other rabbit postoperative body temperature are normal, substantially about 39.3-39.5 ℃.Also have 4 routine rabbit respectively at one of 5 days, 7 days, 17 days, 60 days each death by suffocation of postoperative.This organizes all in various degree concurrent cervical region subcutaneous emphysemas of all rabbit postoperatives, 2 routine concurrent whole body subcutaneous emphysemas wherein, be up to every day from the about 300ml of the subcutaneous gas bleeding in its back, 1 routine rabbit occurred diarrhoea in 14 days after respectively at heeling-in postoperative 5 days and Reconstruction once more, give gentamycin intramuscular injection 10,000 unit/skies, diarrhoea stops after 3 days, and begin gradually to take food, get well, other one routine rabbit 17 days the time, diarrhoea occurs behind Reconstruction, giving gentamycin 10,000 units/sky intramuscular injection suffers from diarrhoea after 2 days and stops, but do not take food, become thin gradually, again through dead after 3 days.Other has the subcutaneous lump of a concurrent cervical region of routine rabbit postoperative, about 2cm diameter, and sphere has smooth capsule.Visible tracheal reconstruction place titanium net surface scar tissue when drawing materials.
The rabbit trachea splanchnoscopy of postoperative death by suffocation in the time of 4 days, as seen the trachea intracavity infects sticking purulent core formation and the red inflammatory granulation tissue growth of reconstruction place, the tube chamber obvious stenosis, other 4 examples are respectively at the rabbit of 5 days, 7 days, 17 days, 60 days each death by suffocation of postoperative, the tangible granulation tissue in various degree of the visible trachea intracavity of trachea splanchnoscopy forms, and tube chamber is very narrow; Check when 4 weeks of postoperative, 8 weeks, 12 weeks that visible trachea intracavity mucosa is rough, tube chamber has in various degree narrow, and the growth of cicatrix and granulation tissue is arranged, and the trachea mucomembranous surface has tangible secretions during 4 weeks.
Postoperative comprises the visible trachea of the rabbit CT examination chamber severe stenosis of death by suffocation, even does not have lacuna fully, all visible tracheal stenosis in various degree of 4 weeks of postoperative, 8 weeks, 12 all trachea CT examination, and AIR STRUCTURE rebuilds and also shows that reconstruction place trachea chamber is narrow.
5 rabbit of hyperplastic tissue's histopathology cut sections for microscopic examination postoperative death by suffocation in the trachea, visible a large amount of granulation tissue hyperplasias, and a small amount of inflammatory cell infiltration; Rebuild operative defect reconstruction place the fibrous scar tissue hypertrophy is arranged, other cells are formed and normal tunica mucosa tracheae no significant difference, mainly contain compositions such as ciliated columnar and goblet cell, mucomembranous surface cilium growing state: postoperative 4 during week the cilium growth of scar tissue surface poor, density is sparse; The scar tissue surface cilium growth of 8 whens week is good slightly, but is worse than the titanium net reconstruction group of fibroin coating; The fibrous tissue layer is than outside the fibrous tissue bed thickness under the titanium net reconstruction group mucosa of fibroin coating under the visible mucosa during 12 weeks, and cilium is also more sparse.
Scanning electron microscope is checked the visible tunica mucosa tracheae cilium of rabbit tube chamber face growing state: postoperative compares with the titanium net reconstruction group of fibroin coating during 4 weeks that cilium is more sparse, short, directivity is poor, self-purification function is poor, and the surface has a little secretions to adhere to; It is good slightly to grow during 8 weeks, and cilium is long, close slightly slightly, but still is worse than the titanium net reconstruction group of fibroin coating, and the complete as yet cilium in subregion covers, and directivity is not good enough; Do not recover normal during 12 weeks yet fully, comparing with the titanium net reconstruction group of normal tunica mucosa tracheae, fibroin coating has certain difference, and directivity not two groups of backs is good, and cilium is shorter than two groups of backs.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102505170A (en) * | 2011-10-26 | 2012-06-20 | 阜宁澳洋科技有限责任公司 | Method for preparing silk protein composite cellulose fibers |
| US8354501B2 (en) | 2005-08-02 | 2013-01-15 | Trustees Of Tufts College | Methods for stepwise deposition of silk fibroin coatings |
| CN103505760A (en) * | 2012-06-29 | 2014-01-15 | 复旦大学附属华山医院 | Airway epithelium-porous silk fibroin protein complex, and preparation method and application thereof |
| CN107182204A (en) * | 2015-08-19 | 2017-09-19 | 翰林大学校产学协力团 | The preparation method for the fluorescence silk protein solution that silk cocoon is extracted is converted from form quality and utilizes its support preparation method |
| CN114699552A (en) * | 2022-02-24 | 2022-07-05 | 武汉亚洲生物材料有限公司 | Preparation method and application of surface composite coating titanium mesh |
| CN114699551A (en) * | 2022-02-24 | 2022-07-05 | 武汉亚洲生物材料有限公司 | Preparation method and application of surface composite coating titanium mesh |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0635515B2 (en) * | 1985-05-31 | 1994-05-11 | 鐘紡株式会社 | Porous silk fibroin powder and method for producing the same |
| DE19950386A1 (en) * | 1999-10-19 | 2001-05-10 | Miladin Lazarov | Biocompatible item |
| CH694935A5 (en) * | 2000-07-26 | 2005-09-30 | Straumann Holding Ag | Oberflaechenmodifizierte implants. |
| WO2003008006A1 (en) * | 2001-07-19 | 2003-01-30 | Dempsey, Donald, J. | Bioactive surface for titanium implants |
| CN1267084C (en) * | 2002-08-01 | 2006-08-02 | 复旦大学 | Medicine releasing control method for implanting type medical equipment |
| WO2004024199A1 (en) * | 2002-09-10 | 2004-03-25 | Scil Technology Gmbh | Metal implant coated under reduced oxygen concentration with osteoinductive protein |
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2005
- 2005-07-01 CN CNB2005100274642A patent/CN100368026C/en not_active Expired - Fee Related
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8354501B2 (en) | 2005-08-02 | 2013-01-15 | Trustees Of Tufts College | Methods for stepwise deposition of silk fibroin coatings |
| CN102505170A (en) * | 2011-10-26 | 2012-06-20 | 阜宁澳洋科技有限责任公司 | Method for preparing silk protein composite cellulose fibers |
| CN102505170B (en) * | 2011-10-26 | 2013-12-18 | 阜宁澳洋科技有限责任公司 | Method for preparing silk protein composite cellulose fibers |
| CN103505760A (en) * | 2012-06-29 | 2014-01-15 | 复旦大学附属华山医院 | Airway epithelium-porous silk fibroin protein complex, and preparation method and application thereof |
| CN103505760B (en) * | 2012-06-29 | 2015-05-20 | 复旦大学附属华山医院 | Airway epithelium-porous silk fibroin protein complex, and preparation method and application thereof |
| CN107182204A (en) * | 2015-08-19 | 2017-09-19 | 翰林大学校产学协力团 | The preparation method for the fluorescence silk protein solution that silk cocoon is extracted is converted from form quality and utilizes its support preparation method |
| CN114699552A (en) * | 2022-02-24 | 2022-07-05 | 武汉亚洲生物材料有限公司 | Preparation method and application of surface composite coating titanium mesh |
| CN114699551A (en) * | 2022-02-24 | 2022-07-05 | 武汉亚洲生物材料有限公司 | Preparation method and application of surface composite coating titanium mesh |
| CN114699551B (en) * | 2022-02-24 | 2023-05-16 | 武汉亚洲生物材料有限公司 | Preparation method and application of surface composite coating titanium mesh |
| CN114699552B (en) * | 2022-02-24 | 2023-05-16 | 武汉亚洲生物材料有限公司 | Preparation method and application of surface composite coating titanium mesh |
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| CN100368026C (en) | 2008-02-13 |
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