CN103800097A - Fibrous membrane for tissue repair and manufacturing method and application thereof - Google Patents
Fibrous membrane for tissue repair and manufacturing method and application thereof Download PDFInfo
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- CN103800097A CN103800097A CN201210457521.0A CN201210457521A CN103800097A CN 103800097 A CN103800097 A CN 103800097A CN 201210457521 A CN201210457521 A CN 201210457521A CN 103800097 A CN103800097 A CN 103800097A
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- fibrous membrane
- tissue repair
- cellosilk
- sticking patch
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Abstract
The invention discloses a fibrous membrane for tissue repair, a manufacturing method thereof, application thereof for the preparation of products for tissue repair, and a product for tissue repair which is made by the fibrous membrane for tissue repair. The fibrous membrane for tissue repair is formed by interweaving cellosilks with the diameter of 10 nanometers-100 micrometers, and is of a porous structure; the filling power is 200-2000 cm<3>/g. According to the fibrous membrane for tissue repair, the conglutination and the proliferation of cells are facilitated, the guide of the cell differentiation is facilitated, the rapid growth of fibrocytes is facilitated, the tight suture between the fibrous membrane and tissue is facilitated, the comfort of a patient is improved, and the occurrence of conditions of patch shrink, infection and conglutination with visceral organs and the like is reduced.
Description
Technical field
The present invention relates to tissue repair fibrous membrane and manufacture method and application, and tissue repair goods.
Background technology
Histoorgan reparation is a worldwide difficult medical problem.In modern surgery operation, often need to repair tissue and organ, as the reparation of hernia repair, the reparation of female pelvic floor functional disorder disease, hard brain/spinal meninges defect repair, nerve trachea defect repair, blood vessel, ligament repair, bone defect repair etc.Due to the particularity of human tissue structure and function, desirable tissue repair need possess following performance with fibrous membrane: 1. certain intensity, can resist mechanical stress, and before not being completed into, wants health tissues to bear Intraabdominal pressure; 2. be convenient to cell adhesion, creep and grow, realize tissue regeneration; 3. preventing that antibacterial from hiding grows, and avoids the infection because using artificial biosynthetic dressing to cause; 4. after implant into body, can keep good dimensional stability, can not shrink and be out of shape; 5. Stability Analysis of Structures, is convenient to be cut into the shape needing and does not decoherence, and is convenient to and tissue apposition; 6. soft, be convenient to molding, increase the operability of operation, reduction patient's sense of discomfort, the effect of raising operation; 7. good biocompatibility, can transmitting tissue grow, and reaches desirable and repairs.
Existing tissue repair fibrous membrane, often there are the following problems:
1) establishment net sheet is most widely used product in fibrous membrane for current tissue repair, but such product surface is coarse, quality is hard, biocompatibility is poor, easily produce foreign body sensation and pain, cause the complication such as common erosion and infection; And the immunological rejection that such material causes is stronger, has more postoperative complication; When it directly contacts with internal organs, organ, easily cause damage, can cause more serious adhesion, cause serious foreign body and immunoreation, need second operation to take out, bring misery to patient, even threat to life.
2) the current electrospinning film of research, commonly occurs that cell is difficult to the slow deficiency of growing into or grow into its inside.
The performance of the tissue repair fibrous membrane that existing braiding as can be seen here,, electrospinning form is not ideal enough.
Summary of the invention
the problem that invention will solve
The present invention makes in view of the problem of above-mentioned prior art.Object is to provide a kind of tissue repair fibrous membrane, it has good mechanical strength, can not repair prerequisite completely for enough mechanics supports, and be conducive to cell adhesion and propagation, guiding cell differentiation, be conducive to the tight stitching between tissue, reduce goods shrinkage tissue repair for, cause infect and with the generation of the situation such as internal organs adhesion, raising patient's comfort, and repair fast, reduce patient's misery.
for the scheme of dealing with problems
Present inventor finds after deliberation, by making tissue repair fibrous membrane have specific loft, can realize above-mentioned purpose, thereby complete the present invention.
Thus, the invention provides a kind of tissue repair fibrous membrane, the cellosilk that described fibrous membrane is 10nm ~ 100 μ m by diameter is interwoven, and has vesicular texture, and its loft is 200 ~ 2000cm
3/ g.
Tissue repair of the present invention can, according to the different cellosilk materials that adopt, select suitable manufacture method to make with fibrous membrane.
Tissue repair of the present invention comprises the steps: by the preferred manufacture method of one of fibrous membrane
(1) by two kinds of cellosilk materials that dissolution properties is different, be dissolved in respectively in corresponding solvent, obtain the cellosilk material solution of two kinds of homogeneous;
(2) two kinds of homogeneous cellosilk material solutions that obtain in step (1) are respectively charged in different electrostatic spinning syringes, the corresponding nozzle needle of two kinds of cellosilk materials is evenly distributed on high-tension electricity source plate, carry out electrostatic spinning, obtain the fibrous membrane of the different crisscross intersection of cellosilk of two kinds of dissolution properties;
(3), according to the dissolution properties of cellosilk material, with suitable solvent, by a kind of cellosilk dissolving in the fibrous membrane of manufacturing in step (2), another remains unchanged, and obtains described tissue repair fibrous membrane.
The another kind of preferable production process of fibrous membrane for tissue repair of the present invention, comprises the steps:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane preparing in step (2) is carried out in predetermined temperature to pre-freeze under the state after solvent supersonic is swelling, then carry out vacuum lyophilization, obtain tissue repair fibrous membrane.
Another preferable production process of fibrous membrane for tissue repair of the present invention, comprises the steps:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane of preparation in step (2) is stretched along the direction of one of the transverse axis of fibrous membrane or longitudinal axis, after stopping stretching, fibrous membrane is shaped under this extended state; Then fibrous membrane is stretched along the direction vertical with above-mentioned draw direction of fibrous membrane, after stopping stretching, fibrous membrane is shaped under this extended state, obtain tissue repair fibrous membrane.
The present invention also provides described tissue repair fibrous membrane in manufacture application in goods for tissue repair.
The present invention further provides the tissue repair goods by the manufacture of described tissue repair fibrous membrane.
The present invention further provides the application for tissue repair of described tissue repair fibrous membrane and tissue repair goods.
the effect of invention
Compared with prior art, the present invention has following beneficial effect:
(1) tissue repair fibrous membrane of the present invention, has bigger serface, be conducive to adhesion and the propagation of cell, and the surface topology of fiber is also conducive to guide cell differentiation;
(2) tissue repair of the present invention has porous fluffy structure with fibrous membrane, is more conducive to fibrocyte grows into rapidly than common electrospinning membrane structure, plays the effect strengthening and fixing, and reaches quick desirable repairing effect;
(3) tissue repair of the present invention is light, soft by fibrous membrane quality, be more conducive to and organize between tight stitching, improve patient's comfort;
(4) tissue repair fibrous membrane of the present invention, can reduce repair reuse artifacts shrinkage, cause infect and with the generation of the situations such as tissue adhesion such as internal organs;
(5) tissue repair of the present invention is not with fibrous membrane owing to can contain living cells composition, and material source is abundant, and cost is lower, avoided the source deficiency of autologous, allosome material and animal derived material, stores transportation simply;
(6) manufacturing approach craft of fibrous membrane of the present invention is simple, and the production time is short, can effectively avoid product in the course of processing to be polluted, and product quality is easy to control, and product standard easily realizes, and product can be realized low cost, high efficiency industrialization is produced.
Accompanying drawing explanation
Fig. 1 is the fluffy fiber film tangent plane scanning electron microscope (SEM) photograph that embodiment 4 obtains;
Fig. 2 is the electrospinning film tangent plane scanning electron microscope (SEM) photograph that embodiment 4 steps (2) obtain;
Fig. 3 is that the fiber of the electrospinning film B that obtains of the fluffy fiber film A that obtains of embodiment 4 and embodiment 4 steps (2) is at 5000 × lower scanning electron microscope comparison diagram;
Fig. 4 is that the fiber of the electrospinning film B that obtains of the fluffy fiber film A that obtains of embodiment 4 and embodiment 4 steps (2) is at 10000 × lower scanning electron microscope comparison diagram;
Fig. 5 is the repairing effect figure of PP group month after operation in embodiment 7;
Fig. 6 is that in embodiment 7, PVDF1 organizes the repairing effect figure of postoperative 6 months;
Fig. 7 is that in embodiment 7, PVDF2 organizes the repairing effect figure of postoperative 6 months;
Fig. 8 is that in embodiment 7, PVDF2 organizes the repairing effect figure of postoperative 6 months;
Fig. 9 is the repairing effect figure that embodiment 7PCU organizes postoperative 6 months;
Figure 10 is the pathology figure that embodiment 9PLLA1 organizes postoperative 6 months;
Figure 11 is the pathology figure that embodiment 9PLLA2 organizes postoperative 6 months;
Figure 12 is animal derived group of pathology figure of postoperative 6 months of embodiment 9;
Figure 13 is the internal anatomy that embodiment 9PLLA1 organizes postoperative 6 months;
Figure 14 is the internal anatomy that embodiment 9PLLA2 organizes postoperative 6 months;
Figure 15 is animal derived group of postoperative internal anatomy of 6 months of embodiment 9.
The specific embodiment
Tissue repair of the present invention is interwoven with the cellosilk that fibrous membrane is 10nm ~ 100 μ m by diameter, has vesicular texture, and its loft is 200 ~ 2000cm
3/ g.
Tissue repair of the present invention is with in fibrous membrane, and described filametntary diameter is preferably 500nm ~ 5 μ m.
Tissue repair of the present invention is preferably 600 ~ 1600cm by the loft of fibrous membrane
3/ g.
Tissue repair of the present invention is preferably 50 ~ 500 milli newton by the pliability of fibrous membrane, is preferably 200 ~ 450 milli newton.
Tissue repair of the present invention is 50 ~ 500 μ m by the preferred average pore size of fibrous membrane.
Tissue repair of the present invention is 0.5 ~ 1.0mm by fibrous membrane preferred thickness.
Tissue repair of the present invention is 20 ~ 80N/cm by the preferred tensile strength of fibrous membrane.
Loft of the present invention refers to the apparent thickness of fibrous membrane and the ratio of surface density 1000 times, that is:
Loft B=apparent thickness T
0/ surface density ω × 10
3
Loft is with cm
3/ g represents, apparent thickness represents with mm, and surface density is with g/m
2represent.Apparent thickness T
0method of testing be to utilize FAST mono-1 compressibility Fabric Style instrument to test according to GB/T 7689.1-2001 method, be expressed as fibrous membrane at 2cN/cm
2thickness under pressure (mm) and fibrous membrane are at 100cN/cm
2thickness under pressure (mm) poor.The test mode of surface density ω is in the thickness situation of ignoring fibrous membrane, measures single the weight under unit are.
Pliability of the present invention refers to that newton represents with milli according to the maximum vector sum of the counter-bending power of diaphragm that in GB/T 8942-2002, method tests out and diaphragm and gap place frictional force, and more the bright film of novel is more soft for softness values.
The diameter of cellosilk described in the present invention is to measure by the method for scanning electron microscope; Described average pore size is with reference to ASTM D 6767-2002, utilizes Capillary Flow pore analysis instrument to measure by bubble point method; Described tensile strength is to measure by GB/T3923.1-1997 " mensuration of fabric ultimate strength and elongation at break " method; Described thickness is measured according to GB/T7689.1-2001 method by compressibility Fabric Style instrument.
Manufacture method as tissue repair of the present invention with fibrous membrane, comprises the method that adopts electrostatic spinning step.
A kind of preferable production process as tissue repair of the present invention with fibrous membrane, can adopt electrostatic spinning technique to carry out in conjunction with dissolution with solvents method, specifically comprises the steps:
(1) by two kinds of cellosilk materials that dissolution properties is different, be dissolved in respectively in corresponding solvent, obtain the cellosilk material solution of two kinds of homogeneous;
(2) two kinds of homogeneous cellosilk material solutions that obtain in step (1) are respectively charged in different electrostatic spinning syringes, the corresponding nozzle needle of two kinds of cellosilk materials is evenly distributed on high-tension electricity source plate, carry out electrostatic spinning, obtain the fibrous membrane of the different crisscross intersection of cellosilk of two kinds of dissolution properties;
(3) according to the dissolution properties of cellosilk material, select suitable solvent that a kind of cellosilk in the fibrous membrane of preparation in step (2) is dissolved, another remains unchanged, and obtains described tissue repair fibrous membrane.
In above-mentioned steps (2), preferably the rate adaptation of micro-injection pump is 0.1 ~ 15.0ml/ hour, more preferably 3 ~ 6ml/ hour.
In above-mentioned steps (2), preferably the voltage-regulation of high tension generator is 5 ~ 45KV, more preferably 20 ~ 36KV.
In above-mentioned steps (2), preferably the receiving range of receiving system is adjusted to 5.0 ~ 30.0cm, more preferably 15.0 ~ 20.0cm.
In the present invention, the different cellosilk material of described two kinds of dissolution properties refers in two kinds of cellosilk materials, to only have wherein a kind of cellosilk material can be dissolved in a certain solvent.Preferred a kind of cellosilk material is wherein: PCU(polycarbonate polyurethane) or PVDF(polyvinylidene fluoride).Preferred another kind of cellosilk material is wherein: PLLA(L-polylactic acid) or PLGA(Poly(D,L-lactide-co-glycolide).
Another kind of preferable production process as tissue repair of the present invention with fibrous membrane, comprises the steps:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane preparing in step (2), under the state after solvent supersonic is swelling, is carried out to pre-freeze under predetermined temperature, then carry out vacuum lyophilization, obtain tissue repair fibrous membrane.
In above-mentioned steps (2), preferably the rate adaptation of micro-injection pump is 0.1 ~ 15.0ml/ hour, more preferably 3 ~ 6ml/ hour.
In above-mentioned steps (2), preferably the voltage-regulation of high tension generator is 5 ~ 45KV, more preferably 30 ~ 36KV.
In above-mentioned steps (2), preferably the receiving range of receiving system is adjusted to 5.0 ~ 30.0cm, more preferably 15.0 ~ 20.0cm.
In above-mentioned steps (3), after the ethanol water that is preferably 50% ~ 95% by the fibrous membrane concentration preparing in step (2) infiltrates, then carry out ultrasonic swelling.
In above-mentioned steps (3), be preferably water for injection for the solvent of ultrasonic swollen fiber film.
In above-mentioned steps (3), after infiltrating in the ethanol water that is preferably 50% ~ 95% by the fibrous membrane preparing in step (2) in concentration, put into the ultrasonic machine that fills water for injection, after ultrasonic swelling 5 ~ 15 minutes, leave standstill, change water for injection, ultrasonic swelling again, repeat operation, until being injected water, ethanol is replaced.Then the fibrous membrane under ultrasonic water for injection solvent swelling state is put into freeze drying box and carried out precooling 2 ~ 5 hours in-50 ℃, then open vacuum and carry out vacuum lyophilization 20 ~ 26 hours, obtain described tissue repair fibrous membrane.
Another preferable production process as tissue repair of the present invention with fibrous membrane, comprises the steps:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane of preparation in step (2) is stretched along the direction of one of the transverse axis of fibrous membrane or longitudinal axis, after stopping stretching, fibrous membrane is shaped under this extended state; Then fibrous membrane is stretched along the direction vertical with above-mentioned draw direction of fibrous membrane, after stopping stretching, fibrous membrane is shaped under this extended state, obtain tissue repair fibrous membrane.
In above-mentioned steps (2), preferably the rate adaptation of micro-injection pump is 0.1 ~ 15.0ml/ hour, more preferably 3 ~ 6ml/ hour.
In above-mentioned steps (2), preferably the voltage-regulation of high tension generator is 5 ~ 45KV, more preferably 30 ~ 36KV.
In above-mentioned steps (2), preferably the receiving range of receiving system is adjusted to 5.0 ~ 30.0cm, more preferably 15.0 ~ 20.0cm.
In above-mentioned steps (3), horizontal and vertical rate of extension is preferably 50 ~ 400mm/min, more preferably 50mm/min ~ 200mm/min independently of one another.
In above-mentioned steps (3), horizontal and vertical tensile elongation is preferably 1.5 ~ 6.0 times of original length independently of one another.
In above-mentioned steps (3), preferably, with the both sides of the fibrous membrane making in clamp clamps above-mentioned steps (2), temperature setting is set to the temperature lower than 0 ℃ ~ 30 ℃ of materials hot deformation temperature, speed with 50 ~ 400mm/min at the uniform velocity stretches, after making fibrous membrane be elongated 1.5 ~ 6.0 times to original length, stop stretching, make at normal temperatures the fibrous membrane 1h ~ 4h that shapes under this extended state, then take off and by the other both sides clamp clamps of fibrous membrane, temperature setting is set to the temperature lower than 0 ℃ ~ 30 ℃ of materials hot deformation temperature, along at the uniform velocity stretching with the speed of 50 ~ 400mm/min with the direction that draw direction is vertical before, after making fibrous membrane be elongated 1.5 ~ 6.0 times to original length, stop stretching, make at normal temperatures the fibrous membrane 1h ~ 4h that shapes under this extended state, obtain described tissue repair fibrous membrane.
The present invention also provides the tissue repair being made by said method fibrous membrane.
Tissue repair of the present invention can be degradation material, non-degradable material or its combination with the filametntary material using in fibrous membrane.Degradation material can be synthesized degradable material or natural macromolecular material; Non-degradable material is preferably poly-fluorine class material, polypropylene based material or polyurethane material.
As degradation material, preferably polylactic acid (PLA) (particularly PLLA (PLLA)), polycaprolactone (PCL), polyglycolic acid (PGA), Poly(D,L-lactide-co-glycolide (PLGA) or 1,3-PD polymer (PDO), collagen protein, gelatin, fibrin, fibroin, the peptide polymer of elastin mimicry, chitosan, modification of chitosan.
As poly-fluorine class material, preferably polyvinylidene fluoride (PVDF) or polytetrafluoroethylene (PTFE).
As polypropylene based material, preferably syndiotactic polypropylene.
As polyurethane material, optimization polyurethane (PU), polycarbonate polyurethane (PCU), polyether based polyurethanes or silicane-modified polyurethane (SPU).
The present invention also provides described tissue repair fibrous membrane in manufacture application in goods for tissue repair.Preferably, tissue repair of the present invention with fibrous membrane for the manufacture of hernia repair sticking patch, female pelvic functional disorder disease repair system (comprising pelvic floor repairing patch or urinary incontinence suspender or fistula patch), artificial bone-head for shoulder's sleeve, hard brain (ridge) film healing sticking patch, artificial skin, pericardium sticking patch, artificial blood vessel, artificial nerve catheter, artificial tooth's film, artificial ligament, artificial heel string, bone reparation (filling) product etc.
The present invention further provides the tissue repair goods by the manufacture of described tissue repair fibrous membrane.
Tissue repair goods of the present invention, are preferably hernia repair sticking patch, female pelvic functional disorder disease repair system (comprising pelvic floor repairing patch or urinary incontinence suspender or fistula patch), artificial bone-head for shoulder's sleeve, hard brain (ridge) film healing sticking patch, artificial skin, pericardium sticking patch, artificial blood vessel, artificial nerve catheter, artificial tooth's film, artificial ligament, artificial heel string, bone reparation (filling) product etc.
The present invention further provides the application for tissue repair of described tissue repair fibrous membrane and tissue repair goods.Preferably provide that described tissue repair fibrous membrane and the tissue repair goods treatment for hernia repair, female pelvic functional disorder disease (including, but not limited to repairing at the bottom of basin or treatment or the fistula reparation of urinary incontinence), shoulder sleeve are repaired, the application of hard brain (ridge) film healing, skin tissue recovering, pericardium reparation, vascular repair, nerve trachea reparation, tooth film healing, ligament repair, heel string reparation, bone reparation (filling).
Further describe the present invention below in conjunction with specific embodiment.The reagent that the embodiment of the present invention adopts and raw material unless stated otherwise, are all commercial conventional products.
Embodiment
Embodiment 1
(1), by polycarbonate polyurethane (PCU), be dissolved in the mixed solvent of DMF and oxolane, the concentration that makes PCU in solution is 12g/100mL, the mixed proportion (volume ratio) of DMF and oxolane is 1:1, obtains the cellosilk material solution of homogeneous.
PLLA is dissolved in hexafluoroisopropanol (HFIP) solution, and making the concentration of PLLA in solution is 5g/100mL, obtains the cellosilk material solution of homogeneous.
(2) the cellosilk material solution of above-mentioned two kinds of homogeneous is respectively charged in two electrostatic spinning syringes, the corresponding nozzle needle of bi-material is evenly distributed on high-tension electricity source plate, regulating the speed of micro-injection pump is 6ml/ hour, regulating the voltage of high tension generator is 22KV, regulating the receiving range of receiving system is 20cm, by bi-material time, static cospinning is prepared the fibrous membrane of the different crisscross intersection of cellosilk material of two kinds of dissolution properties, closes electrostatic spinning after reaching 0.5mm thickness.
(3) by the film taking off, put into hexafluoroisopropanol solvent ultrasonic swelling, dissolve 6 hours, make PLLA material dissolves complete, PCU material remains unchanged, and takes out not dissolved material from solvent, obtains tissue repair fibrous membrane.
In embodiment 1, gained tissue repair is 2 μ m with the average fibre diameter of fibrous membrane, and film thickness is 0.5mm, and average pore size is 350 μ m, and tensile strength is 25N/cm, and loft is 970cm
3/ g, pliability is 250 milli newton.
Embodiment 2
(1) polyvinylidene fluoride (PVDF) is dissolved in to the mixed solvent that volume ratio is DMF/acetone of 4:6, making the concentration of PVDF in solution is 18g/100mL; PLLA (PLLA) is dissolved in hexafluoroisopropanol (HFIP) solution, and making the concentration of PLLA in solution is 5g/100mL, obtains the cellosilk material solution of homogeneous.
(2) the cellosilk material solution of above-mentioned two kinds of homogeneous is respectively charged in two electrostatic spinning syringes, the corresponding nozzle needle of bi-material is evenly distributed on high-tension electricity source plate, regulating the speed of micro-injection pump is 5ml/ hour, regulating the voltage of high tension generator is 22KV, regulating the receiving range of receiving system is 20cm, by bi-material time, static cospinning is prepared the fibrous membrane of the different crisscross intersection of cellosilk material of two kinds of dissolution properties, closes electrostatic spinning after reaching 0.5mm thickness.
(3) by the film taking off, put into hexafluoroisopropanol solvent ultrasonic swelling, dissolve 6 hours, wherein PLLA material is dissolved completely, make tissue repair fibrous membrane.
Described tissue repair is 3 μ m with the average fibre diameter of fibrous membrane, and film thickness is 0.5mm, and average pore size is 450 μ m, and tensile strength is 35N/cm, and loft is 1640cm
3/ g, pliability is 400 milli newton.
Embodiment 3
(1) DMF/acetone mixed solvent that is 4:6 by PVDF material dissolves in volume ratio, making the concentration of PVDF in solution is 18g/100mL; Prepare the cellosilk material solution of homogeneous.
(2) the cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, regulating the speed of micro-injection pump is 6ml/ hour, regulating the voltage of high tension generator is 30KV, regulating the receiving range of receiving system is 20cm, carry out electrostatic spinning and obtain fiber, and fiber is accepted as to membrane structure, and spin after thicknesses of layers is about 0.5mm and close electrostatic spinning, obtain fibrous membrane.
The average fibre diameter of the fibrous membrane herein obtaining is 2 μ m, and film thickness is 0.5mm, and average pore size is 190 μ m, and tensile strength is 43N/cm, and loft is 170cm
3/ g, pliability is 930 milli newton.
(3) complete wetting in the alcoholic solution that is 95% by the fibrous membrane making in step (2) in concentration, then takes out the fibrous membrane after alcoholic solution infiltrates, and puts into the ultrasonic container that fills water for injection, fibrous membrane is fully immersed in water for injection, open ultrasonic, power 90W, ultrasonic 10 minutes, leave standstill after 5 ~ 10 minutes, change the water for injection in ultrasonic container, then open ultrasonic, power 90W, ultrasonic 10 minutes, repeat operation 7 ~ 8 times, replaced complete to ethanol in solution.Then take out the ultrasonic fibrous membrane after swelling of water for injection, be put in the freeze drying box of-50 ℃ and carry out precooling 4 hours, then open vacuum lyophilization, make the fibrous membrane vacuum lyophilization 24 hours of precooling, obtain tissue repair fibrous membrane.
Described tissue repair is 2 μ m with the average fibre diameter of fibrous membrane, and film thickness is 0.6mm, and average pore size is 400 μ m, and tensile strength is 48N/cm, and loft is 1530cm
3/ g, pliability is 420 milli newton.
Embodiment 4
(1) PLLA (PLLA) material is dissolved in hexafluoroisopropanol solvent, making PLLA concentration in solution is 6g/100mL, prepares the cellosilk material solution of homogeneous.
(2) the cellosilk material solution making in step (1) is packed in electrostatic spinning syringe, regulating the speed of micro-injection pump is 6ml/ hour, regulating the voltage of high tension generator is 20KV, regulating the receiving range of receiving system is 15cm, carry out electrostatic spinning and obtain fiber, and fiber is accepted as to membrane structure, and spin after thicknesses of layers is about 0.5mm and close electrostatic spinning, obtain fibrous membrane.
The average fibre diameter of the fibrous membrane herein obtaining is 2 μ m, and film thickness is 0.5mm, and average pore size is 115 μ m, and tensile strength is 33N/cm, and loft is 130cm
3/ g, pliability is 870 milli newton.
(3) then use the both sides of clamp clamps fibrous membrane, be at the uniform velocity to stretch with the speed of 100mm/min under 60 ℃ of conditions in temperature, after making fibrous membrane be elongated 3 times to original length, stop stretching, make at normal temperatures the fibrous membrane 4h that shapes under this extended state, then take off fibrous membrane the other both sides with clamp clamps fibrous membrane, be under 60 ℃ of conditions in temperature, along at the uniform velocity stretching with the speed of 100mm/min with the direction that draw direction is vertical before, after making fibrous membrane be elongated 3 times to original length, stop stretching, make at normal temperatures the fibrous membrane 4h that shapes under this extended state, obtain tissue repair fibrous membrane.
Described tissue repair is 2 μ m with the average fibre diameter of fibrous membrane, and film thickness is 0.6mm, and average pore size is 450 μ m, and tensile strength is 50N/cm, and loft is 1100cm
3/ g, pliability is 400 milli newton.
Embodiment 5
(1) DMF/acetone mixed solvent that is 4:6 by PVDF material dissolves in volume ratio, making the concentration of PVDF in solution is 18g/100mL; Prepare the cellosilk material solution of homogeneous;
(2) the cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, regulating the speed of micro-injection pump is 6ml/ hour, regulating the voltage of high tension generator is 30KV, regulating the receiving range of receiving system is 20cm, carry out electrostatic spinning and obtain fiber, and fiber is accepted as to membrane structure, and spin after thicknesses of layers is about 0.5mm and close electrostatic spinning, obtain fibrous membrane.
The average fibre diameter of the fibrous membrane herein obtaining is 2 μ m, and film thickness is 0.5mm, and average pore size is 190 μ m, and tensile strength is 43N/cm, and loft is 170cm
3/ g, pliability is 930 milli newton.
(3) then use the both sides of clamp clamps fibrous membrane, be at the uniform velocity to stretch with the speed of 100mm/min under 95 ℃ of conditions in temperature, after making fibrous membrane be elongated 2.5 times to original length, stop stretching, make at normal temperatures the fibrous membrane 4h that shapes under this extended state, then take off fibrous membrane the other both sides with clamp clamps fibrous membrane, be under 95 ℃ of conditions in temperature, along at the uniform velocity stretching with the speed of 100mm/min with the direction that draw direction is vertical before, after making fibrous membrane be elongated 2.5 times to original length, stop stretching, make at normal temperatures the fibrous membrane 4h that shapes under this extended state, obtain tissue repair fibrous membrane.
Described tissue repair is 2 μ m with the average fibre diameter of fibrous membrane, and film thickness is 0.6mm, and average pore size is 400 μ m, and tensile strength is 65N/cm, and loft is 1300cm
3/ g, pliability is 450 milli newton.
Embodiment 6
(1) DMF/oxolane mixed solvent that is 1:1 by PCU material dissolves in volume ratio, making the concentration of PCU in solution is 12g/100mL; Prepare the cellosilk material solution of homogeneous;
(2) the cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, regulating the speed of micro-injection pump is 5ml/ hour, regulating the voltage of high tension generator is 36KV, regulating the receiving range of receiving system is 25cm, carry out electrostatic spinning and obtain fiber, and fiber is accepted as to membrane structure, and spin after thicknesses of layers is about 0.5mm and close electrostatic spinning, obtain fibrous membrane.
The average fibre diameter of the fibrous membrane herein obtaining is 2 μ m, and film thickness is 0.5mm, and average pore size is 160 μ m, and tensile strength is 52N/cm, and loft is 110cm
3/ g, pliability is 510 milli newton.
(3) then use the both sides of clamp clamps fibrous membrane, be at the uniform velocity to stretch with the speed of 100mm/min under 80 ℃ of conditions in temperature, after making fibrous membrane be elongated 2.0 times to original length, stop stretching, make at normal temperatures the fibrous membrane 6h that shapes under this extended state, then take off fibrous membrane the other both sides with clamp clamps fibrous membrane, be under 80 ℃ of conditions in temperature, along at the uniform velocity stretching with the speed of 100mm/min with the direction that draw direction is vertical before, after making fibrous membrane be elongated 2.0 times to original length, stop stretching, make at normal temperatures the fibrous membrane 6h that shapes under this extended state, obtain tissue repair fibrous membrane.
Described tissue repair is 2 μ m with the average fibre diameter of fibrous membrane, and film thickness is 0.5mm, and average pore size is 260 μ m, and tensile strength is 60N/cm, and loft is 230cm
3/ g, pliability is 470 milli newton.
The application of embodiment 7 fibrous membranes in hernia repair
By the fibrous membrane making in fibrous membrane, embodiment 5 steps (2) for tissue repair making in embodiment 1, embodiment 5 and clinically use polypropylene mesh (3DMAX
tMthe sample that Mesh is provided by the second Affiliated Hospital of Zhongshan University), be cut into 3.5cm × 6cm, clean, sterilizing, makes respectively PCU hernia repair sticking patch (fibrous membrane by embodiment 1 is manufactured), PVD F1 hernia repair sticking patch (fibrous membrane by embodiment 5 is manufactured), PVDF2 hernia repair sticking patch (by the fibrous membrane manufacture obtaining in embodiment 5 steps (2)) and PP hernia repair sticking patch (being manufactured by polypropylene mesh).
Above-mentioned hernia repair sticking patch is carried out to new zealand rabbit experiment.New zealand rabbit, body weight 2.5-2.8Kg, age 6-12 month, totally 60.Experimental rabbit is divided into 4 groups at random, is respectively PCU group, PVD F1 group, PVDF2 group and PP group, every group of 15 laboratory animals.To entering, the animal of experiment is anaesthetized, preserved skin, lie on the back Baoding on plank; After sterilization paving is single; In abdominal part center, along rabbit hunter's line incision skin, be about 8cm with 4# scalpel, expose muscle, the stomach wall holostrome that is 2cm × 5cm by size (comprising peritoneum, muscle, fascia tissue) excision; PCU group, PVDF1 group, PVDF2 group, PP group adopt respectively PCU hernia repair sticking patch, PVDF1 hernia repair sticking patch, PVDF2 hernia repair sticking patch and PP hernia repair sticking patch, use 0# silk thread by sticking patch and muscle interrupted suture around.4# wire discontinuous sewing skin.
Postoperative animal is carried out to conventional observation and nursing.In observation period, PCU group, PVDF1 group and PVDF2 organize all animal feed water inlets normally, and wound healing is good, has no infection and abdominal part hernia and forms.There is protuberance in 5th ~ 7 days after surgery incision of 5 animals of PP group, 3 animals were in death in postoperative 7th ~ 10 days, and when dissected finds that serious adhesion occurs for sticking patch and internal organs.All the other animals all survived to the time of drawing materials.
Month after operation, organizes every group of 5 zootomies by PCU group, PVDF1 group, PVDF2, and PP organizes 4 zootomies.Touch operative site, PCU group and PVDF1 group operative site touch soft.PVDF2 group touches slightly hard compared with PVDF1 group.Along sticking patch edge, 2cm cuts Musclar layer above, and naked eyes are visible, and PCU group, PVDF1 group and PVDF2 group sticking patch and internal organs are all without adhesion, and blood capillary is grown into centre along sticking patch edge, and cambium covers whole sticking patch substantially.Under mirror, result shows, PCU group, PVDF1 group, PVDF2 group sticking patch visible a large amount of fibroblast and collagen fiber hypertrophy around, minority lymphocytic infiltration (≤25/HPF), more blood capillary proliferation (5-10/HPF).And under mirror, PCU group, PVDF1 group blood capillary is many compared with PVDF2 group, and difference has statistical significance (P<0.05).Visible PCU group sticking patch, PVDF1 group sticking patch, PVDF2 group sticking patch have good histocompatibility; Wherein PCU group sticking patch and the higher loft of PVDF1 group sticking patch are more conducive to tissue and grow into.
PP group touches operative site, and sensation is hard compared with other three groups, can obviously feel the existence of foreign body.Cut skin, the cambium of visible thin layer covers sticking patch surface, but with sticking patch easily openable.Along sticking patch 2cm incision around Musclar layer, the visible PP group sticking patch of naked eyes and the serious adhesion of internal organs, and not easily separated (Fig. 5).Under mirror, result shows, PP group sticking patch inside has in more cavity, cavity has a small amount of fibroblast and collagen to form around, visible small amount blood capillary proliferation (≤2/HPF), a large amount of foreign-body giant cells and lymphocytic infiltration, visible PP group sticking patch histocompatibility is poor, is unfavorable for cell adhesion, rejection and immune inflammation significant reaction.
3 months after operation, PP group is dissected 4 animals, and all the other are organized every group and dissect 5.Cut sticking patch and the tissue within the scope of 2cm around thereof.PCU group, PVDF1 group operative site touch soft, close with autologous tissue.Between sticking patch and skin, be methodically arranged, easily separated, sticking patch and internal organs are without any adhesion, a large amount of cambiums sticking patch inside of growing into.Under mirror, result shows, sticking patch is around combined closely with collagen fiber and fibrocyte; The inner visible a large amount of collagen fiber of sticking patch, with a small amount of blood capillary (3-5/HPF).PVDF2 group material operative site touches more soft, between sticking patch and skin, is methodically arranged, and easily separated, sticking patch and internal organs, without any adhesion, cut sticking patch inside, and cambium is less.Under mirror, result shows, sticking patch is surperficial visible more collagen fiber only, and sticking patch inside has no cambium, and visible cambium is grown into slower to PVDF2 diaphragm inside.
PP group touches operative site, feels harder, can obviously feel the existence of foreign body.The cambium that there is layer on PP group sticking patch surface, lacks unity and coherence with skin, and not easily separated, sticking patch approaches internal organs, with internal organs, serious adhesion occurs, and difficulty or ease are torn, intestinal swelling.Under mirror, result shows, sticking patch has a small amount of fibroblast and collagen to form around, with small amount blood capillary proliferation (≤2/HPF); A large amount of foreign-body giant cells and lymphocytic infiltration, illustrate rejection and immune inflammation significant reaction.
Postoperative 6 months, PP group was dissected 4 animals, and all the other are organized every group and dissect 5.Cut the sticking patch of mend and the tissue within the scope of 2cm around thereof.PVDF1 group is all very soft with the touch of PCU group operative site, and as shown in Fig. 6 and Fig. 9, sticking patch and cambium combine together, more difficult cambium and the sticking patch of distinguishing, between cambium and skin, be methodically arranged, easily separated, cambium and internal organs are without any adhesion.Microscopic observation result shows, implants sticking patch position and is around combined closely with collagen fiber and fibrocyte; Inner visible a large amount of collagen fiber.PVDF2 group operative site touches more soft, as shown in Figure 7 and Figure 8, still can tell sticking patch and cambium, between cambium and skin, is methodically arranged, and easily separated, sticking patch and internal organs are without any adhesion.Microscopic observation result shows, the visible more collagen fiber sticking patch in sticking patch surface, and incision sticking patch inside, cambium is still less.
PP group touches operative site, feels harder, obviously feels the existence of foreign body.PP group lacks unity and coherence with skin, can not separate; Serious with internal organs adhesion; Microscopic observation result shows only around sticking patch, have a small amount of fibroblast and collagen to form, the visible a small amount of blood capillary proliferation (≤5/HPF) in surface, and foreign-body giant cells and lymphocytic infiltration in a large number, foreign body reaction and immunologic rejection are serious; Illustrate that PP group sticking patch is unfavorable for the adhesion of cell and grows into, histocompatibility and repairing effect are poor.
The application of embodiment 8 fibrous membranes in repairing at the bottom of basin
Adopt and make by embodiment 3 pelvic floor repairing patch (PVDF3 sticking patch), embodiment 3 steps 2 that fibrous membrane manufactures and make pelvic floor repairing patch (PVDF4 sticking patch), embodiment 6 that fibrous membrane manufactures and make pelvic floor repairing patch (PCU 1 sticking patch) that fibrous membrane manufactures and pelvic floor repairing patch (PP sticking patch, the Prolift of Johnson & Johnson of polypropylene material manufacture
tM, Guangzhou overseas Chinese hospital provides) and carry out miniature pig experiment.All sticking patch are cut into 2cm × 2cm size.Select 20～25kg, 8 of the female miniature pigs of sexual maturity, PVDF3 group, PVDF4 group, PCU1 group and PP organize totally 4 groups, 2 every group.The miniature pig that enters experiment is implemented to general anesthesia, through hypogastric region stomach wall approach, expose bladder, uterus and vagina epimere.Sticking patch is inserted respectively between vesicovaginal space, and silk thread is fixed.PVDF3 group, PVDF4 group, PCU1 group and the group of PP as a control group adopt respectively PVDF3 sticking patch, PVDF4 sticking patch, PCU1 sticking patch and PP sticking patch.Postoperative animal is carried out to conventional nursing and observation.In all laboratory animal observation periods, all performance is good, and wound healing is good, without implant evict from, the generation such as exposure.Postoperative feed water inlet is normal, and all survival is to the time of drawing materials.
Postoperative 4 weeks, cut the vesicovaginal space tissue specimen including pelvic floor repairing patch.Touch operative site with hands, in PVDF3 group, PVDF4 group, PCU1 sticking patch and PP group, PP material is the hardest, and PVDF4 group material takes second place, and PCU1 organizes material softness, the most soft and the flexible of PVDF3 group material, PVDF3 group, PVDF4 group, the new epithelize of PCU1 group sticking patch surface coverage one deck, not easily openable of sticking patch and cambium, the visible newborn blood capillary of naked eyes, PP group sticking patch surface also covers one deck cambium, and sticking patch easily separates with cambium.Pathological examination shows, there are a large amount of fibroblasts and collagen fiber hypertrophy in PVDF3 group sticking patch inside and surface, and there is more blood capillary (5-10/HPF) sticking patch inside, the visible cambium diaphragm inside of having grown into, and it is very fast to grow into.There are more fibroblast and collagen fiber in PCU1 group, PVDF4 group surface, inner visible a small amount of fibroblast and the collagen fiber hypertrophy of sticking patch, and a small amount of blood capillary proliferation (3-5/HPF), illustrate grow into diaphragm inside but grow into slower of cambium.The inner visible minority lymphocytic infiltration of PVDF4 group, has slight immune inflammation reaction.PP group sticking patch has more fibroblast and collagen around, small amount blood capillary (2-3/HPF), and a small amount of foreign-body giant cell (≤3/HPF), visible cambium cannot grow up to one with diaphragm, repairs slowlyer, and has foreign material repulsion to react.
12 weeks after operation, cuts including repairing vagina and the bladder specimen implant at the bottom of basin.With hands touch, in PVDF3 group, PVDF4 group, PCU1 group and PP group, PP material is the hardest, and PVDF4 material takes second place, and repairing position and surrounding tissue flexibility has compared with significant difference; PCU1 material soft-touch, the most soft and flexible of PVDF3.PVDF3 group sticking patch surface and inside is visible a large amount of fibroblasts and collagen all, and sticking patch and the shape of organizing that grows be integrated, and cannot differentiate and peel off.Microscopic observation result shows, implants sticking patch position and around be combined closely with collagen fiber and fibrocyte, cannot distinguish cambium and diaphragm material, and inside is a large amount of collagen fiber as seen, cambium completely and diaphragm grow up to one, realize Regeneration and Repair.PCU1 group sticking patch is difficult for peeling off with cambium, and naked eyes are visible, and the cambium material internal of growing into, is difficult to differentiation between material and cambium.Microscopic observation result shows, there are a large amount of fibroblasts and a small amount of angiogenesis (3-5/HPF), inner visible small amount blood capillary (2-3/HPF) in surface.PVDF4 group sticking patch and cambium can be peeled off, and there are more fibroblast and a small amount of angiogenesis (3-5/HPF), inner visible small amount blood capillary (≤2/HPF) in surface.PP group sticking patch surface is visible more fibroblast and small amount angiogenesis (2-3/HPF) also, but sticking patch peels off with organizing more easily of growing, and visible cambium cannot grow up to one with diaphragm, and repairing effect is poor.
The loft of visible implantation sticking patch adheres to fast to implant sticking patch surface and enter material internal to cell and organizing and plays an important role, and the high-loft of sticking patch and flexibility is more conducive to cambium and material forms one; Loft is less, and the cambium diaphragm inside of growing into is slower.
The application of embodiment 9 fibrous membranes in cerebral dura mater is repaired
The fibrous membrane making in fibrous membrane, embodiment 4 steps (2) for tissue repair making in embodiment 4 is cut into 4cm × 6cm, clean, sterilizing, make respectively PLLA1 cerebral dura mater and repair sticking patch (fibrous membrane by embodiment 4 is manufactured), PLLA2 cerebral dura mater reparation sticking patch (by the fibrous membrane manufacture obtaining in embodiment 4 steps (2)), matched group adopts the commercial animal derived duramater reparation sheet using clinically (meninges is strong, by Zhongshan University's the 3rd Affiliated Hospital's sampling).
Select 6 of healthy dogs, male and female are not limit, and between body weight 10-15kg, the observation period is 2-3 month.The dog that enters experiment is implemented general anesthesia, Double Tops are opened cranium, artificial bilateral part dura defect and the brain tissue impairment manufactured, then repairs sticking patch with PLLA1 cerebral dura mater and PLLA2 cerebral dura mater reparation sticking patch is being implemented respectively dural repair, 3 dogs of parallel test with an experimental dog brain left and right sides.Repair the animal derived meninges patch of sticking patch and matched group with PLLA2 cerebral dura mater and implement respectively dural repair same the experimental dog brain left and right sides, 3 dogs of parallel test.Postoperative dog is carried out to conventional nursing and observation, when each observation expires, repairing sticking patch position collect specimen, paired observation gross specimen and mirror undertissue.Each experimental animal feeding, to anesthetized animal after the time of setting, exposes skull by the aforementioned cranium mode of opening, and separates to expose and repairs sticking patch outer surface, and vein injects air and puts to death animal, cuts to raise to cut after skull and repairs sticking patch and tissue around.Perusal repairs sticking patch outer surface, quality, with the relation of surrounding tissue, have or not the adhesion situation of cyst, scleroma and inner surface and cerebral tissue.Specimen bottling, is soaked in formalin fixative, and Specimen bottle is labelled.Under room temperature, formalin was fixed after 1 week, got operative site local organization, routine paraffin wax Bao Li, HE tissue section strain.
Rear 6 dogs of performing the operation recover good, and wound healing is good, without secretions.Postoperative feed, water inlet is normal, and the outdoor activities of dog are normal, do not find the dyskinesia.In 3 months posterior veins, push air and put to death dog, centered by operative site, cut specimen being greater than operative site 1cm scope, make it comprise the cerebral tissue of repairing sticking patch and periphery cerebral dura mater and inner face.Cut out after specimen, successively separate brain bone and meninges, the meninges that visible PLLA1 cerebral dura mater is repaired sticking patch implant site is complete, sticking patch implantation place completely fibrous tissue replaces, and do not see embedded material, and implantation place meninges inner surface is without adhesion, corresponding cerebral tissue smooth surface with it, without adhesion.Repairing effect is as dissected as shown in picture Figure 13, and pathological picture as shown in figure 10; The meninges that PLLA2 cerebral dura mater is repaired sticking patch implant site is complete, visible part embedded material has been cracked into fragment, forms new fibrous tissue, and there is a little adhesion on corresponding cerebral tissue surface with it, repairing effect is as dissected as shown in picture Figure 14, and pathological picture as shown in figure 11.And control animals source property meninges is repaired still visible complete not degraded implantation sticking patch of sticking patch implant site, and there is more adhesion in implantation place meninges inner surface and cerebral tissue, can cause the complication such as ectopic discharge, epilepsy as cerebral tissue, repairing effect is as dissected as shown in picture Figure 15, and pathological picture as shown in figure 12.
The application of embodiment 10 fibrous membranes in skin repair
The tissue repair making in embodiment 4 is cut into 5cm × 5cm with fibrous membrane, cleans, sterilizing is made artificial skin and is carried out zoopery.Experimental rabbit body weight 2-2.5kg, age 6-8 month, male and female were not limit, totally 12.Be divided at random two groups (experimental group and matched groups), 6 every group.After the anesthesia of rabbit auricular vein, animal is placed on special-purpose operating table, then standby hair and sterilization.Back holostrome skin is excised with scalpel: excision area is 4*4cm.Then attach artificial dermis: experimental group sticks the artificial skin being made by the fibrous membrane of embodiment 4 in wound site, fix with suture at interval of 1cm.Matched group does blank processing.Experimental group and matched group cover respectively the aseptic oily yarn of one deck and sterile gauze on wound, fix with suture and surrounding skin.
Hands clinical follow animal has or not normal feed, drinking-water, and body temperature, respectively organizes time that covering separates with wound surface and other physiological activity situation, and the reparation situation of observing wound surface.
Postoperative the 2nd, 4,8 weeks every group get 2 rabbits and put to death, get the whole wound surface in back and contiguous normal skin, formalin is fixed, carry out HE dyeing, the newborn situation of organizing in light Microscopic observation corium, inflammatory reaction situation, the regeneration situation of the thickness of epidermal structure and appendages.
Matched group rabbit after surgery all wound surface all with oily yarn or gauze adhesion.Adjuvant has obscission.Wound surface is hemorrhage, has a large amount of tissue fluids to ooze out.Wound surface is linear crust after healing completely.Experimental group postoperative wound surface is dry, and without adhesion, and between wound surface laminating is closely with oily yarn and gauze.Wound is without red and swollen warm, and animal does not occur skin allergy or infects phenomenon.Material covers lower to downright bad phenomenon.Wound is rectangular crust after healing completely.Thicken by the microscopic examination mouthful skin prickle cell layer that heals, basal cell hyperplasia is active, and horny layer, granular layer stratum spinosum and the basal layer of epidermal area are all visible.The attached organ pipe of skin corium, sweat gland, the structures such as hair follicle are visible, and visible a large amount of fibroblasts are parallel to skin surface arranges, more blood capillary proliferation therebetween, local a small amount of lymphocytic infiltration.
Observed result shows: the anti-infection ability of experimental group, prevent that the effect that wound surface is hemorrhage is better than matched group.It is red and swollen, downright bad that experimental group wound surface has no.Histological observation shows that the artificial skin material being made by the fibrous membrane of embodiment 4 promotes skin texture regeneration capacity to be better than matched group.
The application of embodiment 11 fibrous membranes in bone is filled
The tissue repair making in embodiment 2 is cleaned with fibrous membrane, and sterilizing, makes bone repairing support and carries out rabbit zoopery.3 new zealand rabbits, body weight 2.5 ± 0.5Kg, wherein female 1, male 2.Preserved skin after general anesthesia, is placed in animal on special-purpose operating table, and povidone iodine alcohol disinfecting is used in ventricumbent position, completes aseptic dressing, and towel clamp towel forceps fixes.Cut rabbit leg skin, use detacher to separate periosteum, expose tibia hone lamella, with high speed bone drill manufacture rabbit defect of tibial, damaged size is 1cm × 2cm, and the bone repairing support fan-folded being made by the fibrous membrane of embodiment 2 is filled in to damaged place, adjusts implant height, make damaged of itself and bone concordant, sew up.Postoperative 14 days, perusal bone trabecula was thicker, and ultrasonic bone density instrument detects newborn eburnation, and had more woven bone to form.3 months after operation, there is callus formation on damaged surface, bone hole.Knock callus, quality is hard, similar to normal bone tissues hardness, and callus color and autologous bone solid colour.NIP reaction between convalescence.It is good that postoperative animal recovers, and feed water inlet is normal.Extremity motor function is not found the dyskinesia after recovering gradually.
Above embodiment is enough to illustrate that fibrous membrane loft of the present invention is 200cm
3/ g～2000cm
3the beneficial effect that/g brings, can be summarized as follows:
As can be seen from the above table, loft is controlled at 200cm
3/ g～2000cm
3the tissue repairing patch of/g, can form cambium by inducing cell Fast Growth in sticking patch, reaches repairing effect.Can find out equally, for degradation material (PLLA), loft is at 200cm
3/ g～2000cm
3sticking patch within the scope of/g, not compared with the sticking patch within the scope of this, repairing effect is faster with loft, and material degradation is faster.Further, after being the surgical repair of 50 ~ 500 milli Newton range by pliability, the softness relatively of repairing position significantly improves postoperative comfort.
Claims (32)
1. a tissue repair fibrous membrane, is characterized in that, the cellosilk that described fibrous membrane is 10nm ~ 100 μ m by diameter is interwoven, and has vesicular texture, and its loft is 200 ~ 2000cm
3/ g.
2. tissue repair fibrous membrane according to claim 1, is characterized in that, the loft of described fibrous membrane is 600 ~ 1600cm
3/ g.
3. tissue repair fibrous membrane according to claim 1 and 2, is characterized in that, the average pore size of described fibrous membrane is 50 ~ 500 μ m.
4. according to the tissue repair fibrous membrane described in claim 1 ~ 3 any one, it is characterized in that, the thickness of described fibrous membrane is 0.5 ~ 1.0mm, and the tensile strength of described fibrous membrane is 20 ~ 80N/cm.
5. according to the tissue repair fibrous membrane described in claim 1 ~ 4, it is characterized in that, the pliability of fibrous membrane is 50 ~ 500 milli newton.
6. tissue repair fibrous membrane according to claim 5, is characterized in that, the pliability of described fibrous membrane is 200 ~ 450 milli newton.
7. according to the tissue repair fibrous membrane described in claim 1 ~ 6 any one, it is characterized in that, adopt and comprise that the method for electrostatic spinning step makes.
8. tissue repair fibrous membrane according to claim 7, is characterized in that, adopts the method comprising the following steps to make:
(1) by two kinds of cellosilk materials that dissolution properties is different, be dissolved in respectively in corresponding solvent, obtain the cellosilk material solution of two kinds of homogeneous;
(2) two kinds of homogeneous cellosilk material solutions that obtain in step (1) are respectively charged in different electrostatic spinning syringes, the corresponding nozzle needle of two kinds of cellosilk materials is evenly distributed on high-tension electricity source plate, carry out electrostatic spinning, obtain the fibrous membrane of the different crisscross intersection of cellosilk of two kinds of dissolution properties;
(3) according to the dissolution properties of cellosilk material, select suitable solvent that a kind of cellosilk in the fibrous membrane of preparation in step (2) is dissolved, another remains unchanged, and obtains described tissue repair fibrous membrane.
9. tissue repair fibrous membrane according to claim 7, is characterized in that, adopts the method comprising the following steps to make:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane preparing in step (2) is carried out to pre-freeze under the state after solvent supersonic is swelling, then carry out vacuum lyophilization, obtain tissue repair fibrous membrane.
10. tissue repair fibrous membrane according to claim 9, is characterized in that, in above-mentioned steps (3), after the ethanol water that is 50% ~ 95% infiltrates, then carries out ultrasonic swelling by the fibrous membrane concentration preparing in step (2).
11. tissue repair fibrous membranes according to claim 7, is characterized in that, adopt the method comprising the following steps to make:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane of preparation in step (2) is stretched along the direction of one of the transverse axis of fibrous membrane or longitudinal axis, after stopping stretching, fibrous membrane is shaped under this extended state; Then fibrous membrane is stretched along the direction vertical with above-mentioned draw direction of fibrous membrane, after stopping stretching, fibrous membrane is shaped under this extended state, obtain tissue repair fibrous membrane.
12. according to the tissue repair fibrous membrane described in claim 1 ~ 11 any one, it is characterized in that, described filametntary material is degradation material, non-degradable material or its combination; Described degradation material is synthesized degradable material or natural macromolecular material; Described non-degradable material is poly-fluorine class material, polypropylene based material or polyurethane material.
13. tissue repair fibrous membranes according to claim 12, it is characterized in that, described degradation material is polylactic acid, polycaprolactone, polyglycolic acid, Poly(D,L-lactide-co-glycolide or 1,3-PD polymer, collagen protein, gelatin, fibrin, fibroin, the peptide polymer of elastin mimicry, chitosan, modification of chitosan; Described poly-fluorine class material is polyvinylidene fluoride or politef; Described polyurethane material is polyurethane, polycarbonate polyurethane, polyether based polyurethanes or silicane-modified polyurethane.
The method of fibrous membrane for tissue repair described in 14. preparation claim 1 ~ 6 any one, is characterized in that, comprises electrostatic spinning step.
15. methods according to claim 14, is characterized in that, comprise the steps:
(1) by two kinds of cellosilk materials that dissolution properties is different, be dissolved in respectively in corresponding solvent, obtain the cellosilk material solution of two kinds of homogeneous;
(2) two kinds of homogeneous cellosilk material solutions that obtain in step (1) are respectively charged in different electrostatic spinning syringes, the corresponding nozzle needle of two kinds of cellosilk materials is evenly distributed on high-tension electricity source plate, carry out electrostatic spinning, obtain the fibrous membrane of the different crisscross intersection of cellosilk of two kinds of dissolution properties;
(3), according to the dissolution properties of cellosilk material, with solvent, by a kind of cellosilk dissolving in the fibrous membrane of manufacturing in step (2), another remains unchanged, and obtains described tissue repair fibrous membrane.
16. methods according to claim 14, is characterized in that, comprise the steps:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane preparing in step (2) is carried out to pre-freeze under the state after solvent supersonic is swelling, then carry out vacuum lyophilization, obtain tissue repair fibrous membrane.
17. methods according to claim 16, is characterized in that, in above-mentioned steps (3), after the ethanol water that is 50% ~ 95% infiltrates, then carry out ultrasonic swelling by the fibrous membrane concentration preparing in step (2).
18. according to the method described in claim 16 or 17, it is characterized in that, and in above-mentioned steps (3), be water for injection for the solvent of ultrasonic swollen fiber film.
19. methods according to claim 14, is characterized in that, comprise the steps:
(1) cellosilk material is dissolved in solvent, obtains the cellosilk material solution of homogeneous;
(2) the homogeneous cellosilk material solution obtaining in step (1) is packed in electrostatic spinning syringe, carry out electrostatic spinning and obtain fiber, and fiber is received as to membrane structure, obtain fibrous membrane;
(3) fibrous membrane of preparation in step (2) is stretched along the direction of one of the transverse axis of fibrous membrane or longitudinal axis, after stopping stretching, fibrous membrane is shaped under this extended state; Then fibrous membrane is stretched along the direction vertical with above-mentioned draw direction of fibrous membrane, after stopping stretching, fibrous membrane is shaped under this extended state, obtain tissue repair fibrous membrane.
20. methods according to claim 19, is characterized in that, described draft temperature is the temperature lower than 0 ℃ ~ 30 ℃ of cellosilk materials hot deformation temperature.
21. according to the method described in claim 19 or 20, it is characterized in that, the described at the uniform velocity stretching that is stretched as 50 ~ 400mm/min.
22. according to the method described in claim 19 ~ 21 any one, it is characterized in that, length when described fibrous membrane stops stretching is original length 1.5 ~ 6.0 times.
23. according to the method described in claim 19 ~ 22 any one, it is characterized in that, temperature when described fibrous membrane sizing is room temperature.
24. according to the method described in claim 15 ~ 23 any one, it is characterized in that, in the electrostatic spinning of step (2), regulating the speed of micro-injection pump is 0.1 ~ 15.0ml/ hour, regulating the voltage of high tension generator is 5 ~ 45KV, and regulating the receiving range of receiving system is 5.0 ~ 30.0cm.
25. according to the method described in claim 14 ~ 24 any one, it is characterized in that, described filametntary material is degradation material, non-degradable material or its combination; Described degradation material is synthesized degradable material or natural macromolecular material; Described non-degradable material is poly-fluorine class material, polypropylene based material or polyurethane material.
26. methods according to claim 25, it is characterized in that, described degradation material is polylactic acid, polycaprolactone, polyglycolic acid, Poly(D,L-lactide-co-glycolide or 1,3-PD polymer, collagen protein, gelatin, fibrin, fibroin, the peptide polymer of elastin mimicry, chitosan, modification of chitosan; Described poly-fluorine class material is polyvinylidene fluoride or politef; Described polyurethane material is polyurethane, polycarbonate polyurethane, polyether based polyurethanes or silicane-modified polyurethane.
Tissue repair described in 27. claim 1 ~ 13 any one uses fibrous membrane in preparation application in goods for tissue repair.
28. application according to claim 27, is characterized in that described tissue repair goods are hernia repair sticking patch, female pelvic functional disorder disease repair system, artificial bone-head for shoulder's sleeve, cerebral dura mater reparation sticking patch, spinal meninges reparation sticking patch, artificial skin, pericardium sticking patch, artificial blood vessel, artificial nerve catheter, artificial tooth's film, artificial ligament, artificial heel string, bone reparation product.
29. application according to claim 28, is characterized in that described female pelvic functional disorder disease repair system is pelvic floor repairing patch or urinary incontinence suspender or fistula patch.
The 30. tissue repair goods that made with fibrous membrane by the tissue repair described in claim 1 ~ 13 any one.
31. tissue repair goods according to claim 30, described tissue repair goods are hernia repair sticking patch, female pelvic functional disorder disease repair system, artificial bone-head for shoulder's sleeve, cerebral dura mater reparation sticking patch, spinal meninges reparation sticking patch, artificial skin, pericardium sticking patch, artificial blood vessel, artificial nerve catheter, artificial tooth's film, artificial ligament, artificial heel string, bone reparation product.
32. tissue repair goods according to claim 31, is characterized in that described female pelvic functional disorder disease repair system comprises pelvic floor repairing patch or urinary incontinence suspender or fistula patch.
Priority Applications (8)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210457521.0A CN103800097B (en) | 2012-11-14 | 2012-11-14 | A kind of tissue repair fibrous membrane and manufacture method thereof and application |
| IN1362DEN2015 IN2015DN01362A (en) | 2012-11-14 | 2013-11-14 | |
| EP13854718.7A EP2921136B1 (en) | 2012-11-14 | 2013-11-14 | Fiber membranes for repairing tissue and products and preparation method thereof |
| BR112015009502-0A BR112015009502B1 (en) | 2012-11-14 | 2013-11-14 | FIBROUS MEMBRANE, COMPOSITE FIBROUS MEMBRANE, FIBROUS MEMBRANE PREPARATION METHOD, IMPLANTABLE MEMBRANE, IMPLANTABLE MEMBRANE PREPARATION METHOD, IMPLANTABLE MEDICAL DEVICE, ANTI-ADHESION FIBROUS MEMBRANE, FIBROUS MEMBRANE MEMBRANE PREPARATION METHOD |
| JP2015540990A JP6140295B2 (en) | 2012-11-14 | 2013-11-14 | Tissue repair fiber membrane and its product and manufacturing method |
| KR1020157005750A KR101853283B1 (en) | 2012-11-14 | 2013-11-14 | Fibrous Membrane Used for Tissue Repair and Products and Preparation Methods Thereof |
| US14/377,665 US20160045296A1 (en) | 2012-11-14 | 2013-11-14 | Fiber membranes for repairing tissue and products and preparation method thereof |
| PCT/CN2013/001387 WO2014075388A1 (en) | 2012-11-14 | 2013-11-14 | Fiber membranes for repairing tissue and products and preparation method thereof |
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| CN201210457521.0A CN103800097B (en) | 2012-11-14 | 2012-11-14 | A kind of tissue repair fibrous membrane and manufacture method thereof and application |
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Cited By (25)
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