CN104800886A - Gelatin hydrogel myocardium bionic scaffold and preparation method thereof - Google Patents
Gelatin hydrogel myocardium bionic scaffold and preparation method thereof Download PDFInfo
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- CN104800886A CN104800886A CN201510248763.2A CN201510248763A CN104800886A CN 104800886 A CN104800886 A CN 104800886A CN 201510248763 A CN201510248763 A CN 201510248763A CN 104800886 A CN104800886 A CN 104800886A
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Abstract
The invention discloses a gelatin hydrogel myocardium bionic scaffold and a preparation method thereof. Genipin without biological toxicity serves as a cross-linking agent, gelatin hydrogel is subjected to cross-linking modification, and the gelatin hydrogel bionic scaffold is prepared. The cross-linking degree of the scaffold can be further changed by changing the cross-linking conditions of the genipin and the gelatin hydrogel, the elasticity modulus of the scaffold is regulated, the mechanical property of an internal myocardium can be better simulated, appropriate hardness approximate to the internal hardness is provided for the growth of myocardial cells, the growth of the myocardial cells is more facilitated, an ordered spatial structure is formed, and the growth of the myocardial cells and the formation of a heart sheet are promoted.
Description
Technical field
The invention belongs to field of tissue engineering technology, be specifically related to a kind of gelatin hydrogel cardiac muscle biomimetic scaffolds promoting Myocyte growth and cardiac patch to be formed and preparation method thereof.
Background technology
Global disease burden investigation report shows, cardiovascular disease occupy the first in global disease fatality rate, has a strong impact on the Health and Living with harm humans.Ischemic heart desease Global mortality increases year by year, within 1980, has 4,500,000, rises to 5,200,000 in nineteen ninety, within 2000, reaches 6,300,000, and increases to 7,000,000 in 2010 years.Cardiovascular disease brings out multiple serious complication, such as myocardial infarction, then causes disappearance and the death of myocardial necrosis and myocardial cell.The Therapeutic Method of heart disease mainly contains two kinds, and one is carry out heart transplantation, and two is install left ventricular assist device (LVADs).Heart transplantation relate to complicated law and medical procedures, and it is limited to offer organs for transplant; And somewhat expensive needed for LVADs.In addition, some cancer therapy drugs were called back owing to producing serious cardiac toxicity side effect in the last few years, brought huge economic loss to pharmaceuticals, also allowed numerous sufferer deeply hurt simultaneously.Based on above reason, in field of tissue engineering technology, research and development can improve the clinical transplantation material of damaged heart function or myocardium support, and carry out reliable in-vitro evaluation and screening before medicine enters clinical practice, receive benifit greatly by allowing vast sufferer.Find that the mechanical property of timbering material has an impact to myocardial cell pattern and function in cardiac muscle tissue engineering research in recent years, elastic modelling quantity can affect cardiac repair and the vascularization function of support.Cardiac muscular tissue's soft durometer changes with the maturation of individuality, and in newborn individual, cardiac muscular tissue's soft durometer is only at about 1KPa; And in ripe individuality, the early stage myocardium soft durometer of diastole is 10KPa-20KPa, the myocardium soft durometer in relaxing period late period is about 200KPa-300KPa.Therefore the soft durometer of myocardium support controls at 1KPa-300Kpa is good.Study pattern and function that the support showing to have suitable elastic modelling quantity is conducive to heart cell, the percentage elongation of cell is the highest, and excited threshold value is more reasonable, and the contraction dynamics of generation is the strongest.In addition, in Tissue Engineering Study, find the bootable cell distribution of the regulation and control of space structure, promote maturation and the propagation of cell.Nearest research display: the hydrogel close with body heart muscle layer soft durometer can promote growth and the stem cell myocardiac differentiation of myocardial cell in testing in vitro, can improve cardiac function in vivo in animal heart infarction model.Synthetic water gel and natural hydrogel are extensively studied in cardiac muscle tissue engineering.But the poor and degradation speed of synthetic water gel biological compatibility is slow, is unfavorable for that later phase clinical is applied.Natural hydrogel wide material sources, have good biocompatibility and the sensitivity to environment, and mostly have good biological degradability.Gelatin good biocompatibility, nontoxic, non-immunogenicity, biodegradable, receive much concern in biomaterial and biomedical applications.But not modified gelatin stability and bad mechanical property, the performance of gelatin hydrogel is improved further by crosslinked method.Chemical crosslinking often uses the cross-linking agent such as formaldehyde, glutaraldehyde, carbodiimide to carry out cross-linking modified to gelatin at present, but these three kinds of cross-linking agent exist the problem of bio-toxicity, the biocompatibility of the gelatin after impact is crosslinked.Genipin is that the geniposide in cape jasmine fruit obtains after beta-glucosidase enzyme hydrolysis.Genipin and geniposide be Chang Zuowei anti-inflammatory agent and choleretic in Chinese medicine.In recent years, also there are research and utilization genipin and gelatin to be cross-linked, improve gelatin performance.The people such as Lien have studied three kinds of different crosslinking methods to the impact of gelatin hydrogel, the cryodesiccated order of main research is on the impact of gelatin hydrogel microstructure, but it cannot realize regulation and control (the Lien S-M to gelatin hydrogel soft durometer, Li W-T, Huang T-J.Genipin-crosslinked gelatin scaffolds for articular cartilage tissue engineering with anovel crosslinking method [J] .Materials Science and Engineering:C, 2008, 28 (1): 36-43.).The people such as Liang have studied carbodiimide, the bio-toxicity of the gelatin that genipin is cross-linked, find the gelatin hydrogel that the gelatin hydrogel that genipin is cross-linked is cross-linked lower than carbodiimide, but its research is conceived to the application of gelatin hydrogel in medicament slow release, but not myocardium support (Liang H C, Chang W H, Liang H F, etal.Crosslinking structures of gelatin hydrogels crosslinked with genipin or a water ?soluble carbodiimide [J] .Journal of Applied Polymer Science, 2004, 91 (6): 4017-4026.).The people such as Zhang Pengyun have studied the impact of genipin on the modification of Static Spinning glutin nano fabric film, but the gelatine nano fiber soft durometer that electrospinning process obtains is too large, be not suitable as the material (Zhang Pengyun of myocardium support, Zhang Jiansong, Xu Xiaohong etc. Biological cross-linker genipin is on the impact [J] of Static Spinning glutin nano fabric film modification. Chinese Tissue Engineering Study and clinical rehabilitation, 2009,13 (8): 1500-1504.).
Summary of the invention
The problem of Myocyte growth is not suitable in order to solve cross-linking agent bio-toxicity and the elastic modelling quantity after being cross-linked in prior art, the present invention utilizes genipin cross-linked gelatin hydrogel, determine the parameters such as the concentration of genipin, crosslinking temperature, solvent, obtain to organize with body myocardium soft durometer consistent, be beneficial to the gelatin hydrogel support that Myocyte growth and cardiac patch formed.Another object of the present invention is also to provide a kind of preparation method promoting the gelatin hydrogel support that Myocyte growth and cardiac patch are formed.
In order to reach goal of the invention, the technical solution used in the present invention is: the gelatin hydrogel in described biomimetic scaffolds is cross-linked through genipin.
Described biomimetic scaffolds surface has the micro structure of bootable myocardial cell high-sequential aligned growth.
Described micro structure is groove structure arranged in parallel.
The ditch of described groove structure and the width of ridge are 5 μm ~ 25 μm, and the difference in height of ditch and ridge is 0.5 μm ~ 1 μm.
The ditch of described groove structure and the width of ridge are preferably 10 μm ~ 20 μm.
The ditch of described micro structure and the width of ridge are more preferably 15 μm.
In described biomimetic scaffolds, the mass volume ratio of gelatin is 6% ~ 20%.
The detailed process of the preparation method of described biomimetic scaffolds is as follows: first that gelatin is soluble in water, obtains the gelatin solution that mass volume ratio is 6% ~ 20%; Then draw gained gelatin solution plastic, the gelatin hydrogel of surface without micro structure can be obtained; Finally by genipin dissolution with solvents, obtain the genipin solution that mass volume ratio is 0.1% ~ 1.0%, again the described gelatin hydrogel without micro structure is soaked in described genipin solution, 4 hours ~ 72 hours are cross-linked at being placed in 5 DEG C ~ 30 DEG C, after crosslinked, clean support with ultra-pure water, finally prepare described biomimetic scaffolds; The described solvent for dissolving genipin be water or concentration be 10mM and pH be 7.4 HEPES buffer.
The temperature of preparation method Zhong jing Buddhist nun's usual friendship connection of described biomimetic scaffolds is 25 DEG C.
The concrete preparation process that described surface has the biomimetic scaffolds of micro structure is as follows: first that gelatin is soluble in water, obtains the gelatin solution that mass volume ratio is 6% ~ 20%; Then the micro structure in silicon template is transferred in PDMS template, then gets institute's gelatine solution and drop to PDMS template surface plastic, after gelatin hydrogel plastic, it is peeled off from PDMS template, namely prepare the gelatin hydrogel that surface has micro structure; Finally, by genipin dissolution with solvents, obtain the genipin solution that mass volume ratio is 0.1% ~ 1.0%, the described gelatin hydrogel with micro structure is soaked in described genipin solution, be cross-linked 4 hours ~ 72 hours at being placed in 5 DEG C ~ 30 DEG C, after crosslinked, clean support with ultra-pure water, the genipin that removing is residual, namely prepares the biomimetic scaffolds that described surface has micro structure; The described solvent for dissolving genipin be water or concentration be 10mM and pH be 7.4 HEPES buffer.
The temperature that described surface has preparation method Zhong jing Buddhist nun's usual friendship connection of the biomimetic scaffolds of micro structure is 25 DEG C.
Compared with prior art, beneficial effect of the present invention is embodied in:
(1) construct a kind of gelatin hydrogel support crosslinked through genipin of inanimate object toxicity and in cardiac muscle tissue engineering technical field, solve the problem of the bio-toxicity of cross-linking agent residual in prior art.
(2) gelatin hydrogel support of the present invention can by changing the elastic modelling quantity of cross linking conditions regulation and control hydrogel scaffold, reach the effect that mechanics is controlled, more can the mechanical property of analogue body heart muscle layer, the Integral synchronous that earlier can realize myocardial cell is beated and drives Substrate Contraction, thus promotes that Myocyte growth and cardiac patch are formed.
(3) gelatin hydrogel support of the present invention can be grown along micro structure direction high-sequential by regulation and control hydrogel surface microstructure directs myocardial cell, improve the percentage elongation of cell, the contraction dynamics that myocardial cell is produced increases, and promotes the formation of Myocyte growth and cardiac patch further.
Accompanying drawing explanation
Fig. 1: be the situation of beating of neonatal cardiac myocytes on the gelatin hydrogel biomimetic scaffolds that embodiment 2 prepares, I, II and III are diverse location on support.
Detailed description of the invention:
In order to deepen the understanding of the present invention, below in conjunction with embodiment, the invention will be further described.
Embodiment 1:
The preparation of step 1, gelatin solution: the gelatin taking 0.6g is dissolved in 10mL ultra-pure water, after the swelling 30min of room temperature, is placed in 40 DEG C of water-baths and dissolves, and forming mass volume ratio is the gelatin solution of 6%;
Step 2, preparation without the gelatin hydrogel support of micro structure: it is the cylinder mold of 10mm that the gelatin solution drawn in 400 μ L steps 1 injects internal diameter, be placed in 4 DEG C of environment and within 2 hours, can form gelatin hydrogel support without micro structure.
Step 3, genipin cross-linked hydrogel: take a certain amount of genipin and be dissolved in ultra-pure water, forming mass volume ratio is the genipin solution of 0.1%.The above-mentioned hydrogel scaffold prepared is soaked in genipin solution, is placed in the crosslinked 4h of 5 DEG C of incubators.After crosslinked, ultra-pure water cleaning support three times, the genipin that removing is residual, namely prepares gelatin hydrogel support.
Embodiment 2:
The preparation of step 1, gelatin solution: the gelatin taking 0.6g is dissolved in 10mL ultra-pure water, after the swelling 30min of room temperature, is placed in 60 DEG C of water-baths and dissolves, and forming mass volume ratio is the gelatin solution of 6%;
Step 2, preparation without the gelatin hydrogel support of micro structure: it is the cylinder mold of 10mm that the gelatin solution drawn in 400 μ L steps 1 injects internal diameter, be placed in 10 DEG C of environment and within 2 hours, can form gelatin hydrogel support without micro structure.
Step 3, genipin cross-linked hydrogel: take a certain amount of genipin and be dissolved in HEPES buffer (10mM, pH 7.4), forming mass volume ratio is the genipin solution of 0.2%.The above-mentioned hydrogel scaffold prepared is soaked in genipin solution, is placed in the crosslinked 24h of 25 DEG C of incubators.After crosslinked, ultra-pure water cleaning support three times, the genipin that removing is residual, namely prepares gelatin hydrogel support.
Embodiment 3:
The preparation of step 1, gelatin solution: the gelatin taking 0.6g is dissolved in 10mL ultra-pure water, after the swelling 30min of room temperature, is placed in 60 DEG C of water-baths and dissolves, and forming mass volume ratio is the gelatin solution of 6%;
Step 2, preparation without the gelatin hydrogel support of micro structure: it is the cylinder mold of 10mm that the gelatin solution drawn in 400 μ L steps 1 injects internal diameter, be placed in 4 DEG C of environment and within 2 hours, can form gelatin hydrogel support without micro structure.
Step 3, genipin cross-linked hydrogel: take a certain amount of genipin and be dissolved in HEPES buffer (10mM, pH 7.4), forming mass volume ratio is the genipin solution of 0.8%.The above-mentioned hydrogel scaffold prepared is soaked in genipin solution, is placed in the crosslinked 24h of 25 DEG C of incubators.After crosslinked, ultra-pure water cleaning support three times, the genipin that removing is residual, namely prepares gelatin hydrogel support.
Embodiment 4:
The preparation of step 1, gelatin solution: the gelatin taking 2g is dissolved in 10mL ultra-pure water, after the swelling 30min of room temperature, is placed in 90 DEG C of water-baths and dissolves, and forming mass volume ratio is the gelatin solution of 20%;
Step 2, preparation without the gelatin hydrogel support of micro structure: it is the cylinder mold of 10mm that the gelatin solution drawn in 400 μ L steps 1 injects internal diameter, be placed in 4 DEG C of environment and within 2 hours, can form gelatin hydrogel support without micro structure.
Step 3, genipin cross-linked hydrogel: take a certain amount of genipin and be dissolved in HEPES buffer (10mM, pH 7.4), forming mass volume ratio is the genipin solution of 1.0%.The above-mentioned hydrogel scaffold prepared is soaked in genipin solution, is placed in the crosslinked 72h of 30 DEG C of incubators.After crosslinked, ultra-pure water cleaning support three times, the genipin that removing is residual, namely prepares gelatin hydrogel support.
Embodiment 5:
Make micro structure gelatin hydrogel: the host and the firming agent that first take PDMS, take mass ratio as the ratio mixing of 10:1, be uniformly mixed with spoon and be placed in vacuum drying oven, evacuation 15min, the bubble in removing reagent.Subsequently, PDMS is poured into the silicon template surface that surface has micro structure, puts into 60 DEG C of baking oven 2h, the micro structure in wherein said silicon template is groove structure arranged in parallel, and the width of ditch and ridge is 5 μm, and the difference in height of ditch and ridge is 0.5 μm.After PDMS solidification, it is peeled off from silicon template surface, namely prepares the PDMS template (micro structure in PDMS template is contrary with the micro structure in silicon template) that can reuse.During preparation micro structure hydrogel, get 30 μ L hydrogel solutions and drop to PDMS template surface, slide is gently buckled in hydrogel solution surface, in 4 DEG C of environment, leave standstill 24 hours plastics.After hydrogel plastic is air-dry, it is peeled off from PDMS template, namely prepare the micro structure gelatin hydrogel consistent with silicon template micro structure.
The method of employing genipin cross-linked gelatin hydrogel is afterwards identical with embodiment 1.
Embodiment 6:
Make micro structure gelatin hydrogel and embodiment 5 uniquely unlike: when preparing micro structure hydrogel in 25 DEG C of environment standing 72 hours plastics.
The method of employing genipin cross-linked gelatin hydrogel is afterwards identical with embodiment 1.
Embodiment 7:
Make micro structure gelatin hydrogel and embodiment 5 unique unlike: the ditch of the micro structure in silicon template and the width of ridge are 10 μm, and the difference in height of ditch and ridge is 0.5 μm.
The method of employing genipin cross-linked gelatin hydrogel is afterwards identical with embodiment 2.
Embodiment 8:
Make micro structure gelatin hydrogel and embodiment 5 unique unlike: the ditch of the micro structure in silicon template and the width of ridge are 15 μm, and the difference in height of ditch and ridge is 1 μm.
The method of employing genipin cross-linked gelatin hydrogel is afterwards identical with embodiment 3.
Embodiment 9:
Make micro structure gelatin hydrogel and embodiment 5 unique unlike: the ditch of the micro structure in silicon template and the width of ridge are 20 μm, and the difference in height of ditch and ridge is 1 μm.
The method of employing genipin cross-linked gelatin hydrogel is afterwards identical with embodiment 4.
Embodiment 10:
Make micro structure gelatin hydrogel and embodiment 5 unique unlike: the ditch of the micro structure in silicon template and the width of ridge are 25 μm, and the difference in height of ditch and ridge is 1 μm.
The method of employing genipin cross-linked gelatin hydrogel is afterwards identical with embodiment 4.
Embodiment 11:
The biomimetic scaffolds that embodiment 1-10 prepares utilizes atomic force microscope (Nanosurf company of Switzerland) to measure its elastic modelling quantity, the elastic range recorded is 1.3 ± 0.21KPa ~ 289 ± 10.3KPa, belong to the soft durometer scope of cardiac muscular tissue when individual body maturation and contraction diastole, be therefore conducive to the growth of myocardial cell.As shown in table 1.
The elastic modelling quantity of the biomimetic scaffolds of the different embodiment of table 1.
Embodiment 12:
When adopting neonatal cardiac myocytes to cultivate the 6th day on the gelatin hydrogel biomimetic scaffolds that embodiment 2 prepares, culture fluid is removed, cleans three times with tyrode, the culture fluid that removing is residual.Subsequently, the Fluo-4AM Ca of freshly prepared 2 μMs is added
2+indicator working solution, is placed in 37 DEG C containing 5%CO
220min is hatched in incubator.Remove Fluo-4AM Ca afterwards
2+indicator working solution, tyrode adds culture fluid after cleaning three times, is placed in 37 DEG C of incubators and hatches 30min.The calcium ionic current situation of change of myocardial cell is observed under sample being placed in fluorescence inverted microscope, and capture video.With fluorescent brightness in Image J software statistics video over time.The change of calcium ionic current can the bioelectrical activity of reflecting myocardium cell, and at every turn beating of myocardial cell all pumps into cell along with calcium ion and pump cell.Fig. 1 is the calcium ion fluorescent brightness curve chart over time of diverse location place on support (I, II and III, random selecting) neonatal cardiac myocytes.
Claims (10)
1. a gelatin hydrogel cardiac muscle biomimetic scaffolds, is characterized in that the gelatin hydrogel in described biomimetic scaffolds adopts genipin to be cross-linked.
2. biomimetic scaffolds according to claim 1, is characterized in that described biomimetic scaffolds surface has the micro structure of bootable myocardial cell high-sequential aligned growth.
3. biomimetic scaffolds according to claim 2, is characterized in that described micro structure is groove structure arranged in parallel.
4. biomimetic scaffolds according to claim 3, is characterized in that the ditch of described groove structure and the width of ridge are 5 μm ~ 25 μm, and the difference in height of ditch and ridge is 0.5 μm ~ 1 μm.
5., according to the biomimetic scaffolds described in claim 4, it is characterized in that the ditch of described groove structure and the width of ridge are 10 μm ~ 20 μm.
6., according to the biomimetic scaffolds described in claim 5, it is characterized in that the ditch of described micro structure and the width of ridge are 15 μm.
7., according to described biomimetic scaffolds arbitrary in claim 1-6, it is characterized in that the mass volume ratio of gelatin in biomimetic scaffolds is 6% ~ 20%.
8. a preparation method for biomimetic scaffolds as claimed in claim 1, is characterized in that concrete preparation process is as follows: first that gelatin is soluble in water, obtains the gelatin solution that mass volume ratio is 6% ~ 20%; Then draw gained gelatin solution plastic, the gelatin hydrogel of surface without micro structure can be obtained; Finally by genipin dissolution with solvents, obtain the genipin solution that mass volume ratio is 0.1% ~ 1.0%, the more described gelatin hydrogel without micro structure is soaked in described genipin solution, be placed in 5
oc ~ 30
obe cross-linked 4 hours ~ 72 hours under C, after crosslinked, clean support with ultra-pure water, finally prepare described biomimetic scaffolds; The described solvent for dissolving genipin be water or concentration be 10mM and pH be 7.4 HEPES buffer.
9., as the surface as described in arbitrary in claim 2-6 has a preparation method for the biomimetic scaffolds of micro structure, it is characterized in that concrete preparation process is as follows: first that gelatin is soluble in water, obtain the gelatin solution that mass volume ratio is 6% ~ 20%; Then the micro structure in silicon template is transferred in PDMS template, then gets institute's gelatine solution and drop to PDMS template surface plastic, after gelatin hydrogel plastic, it is peeled off from PDMS template, namely prepare the gelatin hydrogel that surface has micro structure; Finally, by genipin dissolution with solvents, obtain the genipin solution that mass volume ratio is 0.1% ~ 1.0%, the described gelatin hydrogel with micro structure is soaked in described genipin solution, is placed in 5
oc ~ 30
obe cross-linked 4 hours ~ 72 hours under C, after crosslinked, clean support with ultra-pure water, the genipin that removing is residual, namely prepare the biomimetic scaffolds that described surface has micro structure; The described solvent for dissolving genipin be water or concentration be 10mM and pH be 7.4 HEPES buffer.
10. the preparation method of biomimetic scaffolds according to claim 8, is characterized in that the temperature that genipin is cross-linked is 25
oc.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105727365A (en) * | 2016-03-29 | 2016-07-06 | 王嘉显 | Heart band-aid and preparation method thereof |
| CN105749350A (en) * | 2016-04-21 | 2016-07-13 | 四川大学 | Myocardial patch and preparation method thereof |
| CN107308504A (en) * | 2017-06-17 | 2017-11-03 | 常州帝君金属构件厂 | A kind of preparation method of myocardium biomimetic scaffolds |
| CN109464664A (en) * | 2018-11-13 | 2019-03-15 | 南昌大学 | It is a kind of for alleviating the preparation method of the intelligent eye sticker of dry eye condition |
| CN113292743A (en) * | 2021-05-21 | 2021-08-24 | 福州大学 | Injectable high-pressure-resistant high-strength anti-freezing genipin crosslinked gelatin hydrogel and preparation method thereof |
| CN113999426A (en) * | 2021-12-21 | 2022-02-01 | 浙江理工大学 | Preparation method of porous structure double-layer gelatin hydrogel driver |
| CN116370698A (en) * | 2023-06-06 | 2023-07-04 | 天津医科大学眼科医院 | Hydrogel scleral plug and preparation method and application thereof |
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| CN105727365B (en) * | 2016-03-29 | 2018-08-28 | 南京艾尔普再生医学科技有限公司 | Heart adhesive bandage and preparation method thereof |
| CN105727365A (en) * | 2016-03-29 | 2016-07-06 | 王嘉显 | Heart band-aid and preparation method thereof |
| CN105749350B (en) * | 2016-04-21 | 2019-05-21 | 四川大学 | A kind of cardiac muscle sticking patch and preparation method thereof |
| CN105749350A (en) * | 2016-04-21 | 2016-07-13 | 四川大学 | Myocardial patch and preparation method thereof |
| CN107308504A (en) * | 2017-06-17 | 2017-11-03 | 常州帝君金属构件厂 | A kind of preparation method of myocardium biomimetic scaffolds |
| CN109464664B (en) * | 2018-11-13 | 2021-07-20 | 南昌大学 | Preparation method of intelligent eye patch for relieving xerophthalmia symptoms |
| CN109464664A (en) * | 2018-11-13 | 2019-03-15 | 南昌大学 | It is a kind of for alleviating the preparation method of the intelligent eye sticker of dry eye condition |
| CN113292743A (en) * | 2021-05-21 | 2021-08-24 | 福州大学 | Injectable high-pressure-resistant high-strength anti-freezing genipin crosslinked gelatin hydrogel and preparation method thereof |
| CN113292743B (en) * | 2021-05-21 | 2022-08-19 | 福州大学 | Injectable high-pressure-resistant high-strength anti-freezing genipin crosslinked gelatin hydrogel and preparation method thereof |
| CN113999426A (en) * | 2021-12-21 | 2022-02-01 | 浙江理工大学 | Preparation method of porous structure double-layer gelatin hydrogel driver |
| CN113999426B (en) * | 2021-12-21 | 2022-11-25 | 浙江理工大学 | Preparation method of porous structure double-layer gelatin hydrogel driver |
| CN116370698A (en) * | 2023-06-06 | 2023-07-04 | 天津医科大学眼科医院 | Hydrogel scleral plug and preparation method and application thereof |
| CN116370698B (en) * | 2023-06-06 | 2023-08-08 | 天津医科大学眼科医院 | Hydrogel scleral plug and preparation method and application thereof |
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Application publication date: 20150729 |