A multifunctional heart attachment support device, a pericardial fluid collection system and a collection method

Abstract

The present application belongs to the technical field of medical devices, and discloses a multifunctional heart attaching and supporting device, a pericardial fluid collection system and a collection method. The multifunctional heart attaching and supporting device mainly collects pericardial fluid through the biomimetic suction cup structure on the inner side and can also realize in-situ drug delivery for the heart. The pericardial fluid collection system and collection method provided by the present application can collect pericardial fluid through the device provided by the present application, and then perform biochemical detection. The system and method can take into account early physical auxiliary treatment and late drug delivery, and the drug delivery is more localized and accurate. In addition, the system and method can also monitor the disease development and feedback the treatment effect through pericardial fluid, form a "monitoring-treatment-feedback" closed loop, realize early intervention and early treatment, and make the treatment of heart failure no longer limited to simple drug treatment and physical constraint treatment.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of medical devices, in particular to a multifunctional heart attachment support device, a pericardial fluid collection system and a collection method. BACKGROUND

[0002] Heart failure, referred to as heart failure, is a syndrome caused by primary heart damage under normal venous return, resulting in reduced cardiac output and inability to meet tissue metabolic needs. After heart failure occurs, the heart pump function is insufficient and the cardiac output is reduced, and the ventricle is thickened in the late stage of remodeling, and the cardiac output cannot meet the normal activity of the body. It is manifested as decreased exercise tolerance, and even normal activities will cause chest tightness, shortness of breath, fatigue and other blood supply insufficiency symptoms once the distance is slightly long or the duration is slightly long, which seriously affects the quality of life of patients.

[0003] For early heart failure, the commonly used treatment method is oral systemic administration. The new drug delivery method currently studied in the laboratory stage, such as a one-time drug delivery cardiac hydrogel patch, can store therapeutic drugs or cells, and release drugs or cells locally after implantation, thereby achieving a therapeutic effect. For example, a cardiac microneedle patch, which uses cardiac stromal cells as a matrix, can promote myocardial regeneration when transplanted to the damaged heart wall (Tang, J., Wang, J., Huang, K., Ye, Y., Su, T., Qiao, L., Hensley, M. T., Caranasos, T. G., Zhang, J., Gu, Z., & Cheng, K. (2018). Cardiac cell-integrated microneedle patch for treating myocardial infarction. Science advances, 4(11), eaat9365. https: / / doi.org / 10.1126 / sciadv.aat9365). The disadvantage is that a thoracotomy is required for each drug delivery. However, the cardiac patch requires a thoracotomy for each drug delivery implantation, which causes great damage and is limited to single emergency treatment.

[0004] For end-stage heart failure, common treatment methods include heart transplantation, or the use of ventricular assist devices combined with drug-assisted therapy. The most common and currently clinically used implantable long-term left heart assist device is the blood pump type ventricular assist device (LVAD), such as the domestic Chongqing Yongren heart and imported HeartMate. The blood pump is composed of a pump body, a pipeline and a power system, which connects the left ventricle and the aorta, and transmits left ventricular blood to the aorta through the main working mechanism, thereby reducing the burden on the heart and improving body perfusion to "treat" end-stage heart failure. However, due to its blood-contacting treatment mechanism, patients need to take anticoagulant drugs for a long time, but long-term machine life still has a chance of thrombosis, infection and other complications, which undoubtedly increases the patient's economic burden and reduces the patient's quality of life. Laboratory stage research and development of ventricular assist devices include non-blood contact devices such as soft robotic sleeve devices (Soft Robotic Sleeve), which mainly include: air pressure transmission device, air pump, soft sleeve, the above-mentioned devices mainly use compressed air to drive the soft sleeve to compress and rotate, and then simulate the normal heart movement (Roche, E. T., Horvath, M. A., Wamala, I., Alazmani, A., Song, S. E., Whyte, W., Machaidze, Z., Payne, C. J., Weaver, J. C., Fishbein, G., Kuebler, J., Vasilyev, N. V., Mooney, D. J., Pigula, F. A., & Walsh, C. J. (2017). Soft robotic sleeve supports heart function. Science translational medicine, 9(373), eaaf3925. https: / / doi.org / 10.1126 / scitranslmed.aaf3925), although it does not contact with blood, but it is still the same as other ventricular assist devices in function, and it is difficult to combine with other heart failure treatment methods, and can only be used for physical support treatment, which has obvious single and limitation in use, especially cannot be directly monitored and intervened by biology and drugs. For example, after heart failure occurs, the changes in pericardial fluid are closely related to the state of the heart. In the pathological process of the heart, the levels of various active biochemical factors in the pericardial fluid change significantly, including hormones, neurotransmitters, cytokines, and local growth and apoptosis factors.However, the current method for obtaining pericardial fluid is generally puncture collection, and the pericardial fluid obtained by this collection method cannot avoid the mixing of other body fluids in the chest cavity, and it is difficult to form an accurate feedback mechanism of the change of the composition of the pericardial fluid and the drug. In the field of drug treatment for heart failure, pericardial fluid also needs to be collected for component analysis to establish the correlation between the relevant components in the pericardial fluid and the drug. The lack of the above-mentioned accurate feedback mechanism also restricts the progress of the research and development of drugs for heart failure.

[0005] The inventors of the present application have previously disclosed a patent document CN107049232B, which provides an attached heart function monitoring and / or intervention system, including a heart support device and a heart function monitoring device and / or an intervention device. The heart support device is coated on the outer surface of the ventricle and / or atrium or is supported and attached to the inner surface of the heart chamber. The physiological and biochemical sensor is connected to the heart function monitoring device. The physiological and biochemical sensor detects or senses the change of the physiological and biochemical parameters of the inner or outer surface of the heart, and transmits the change to the heart function monitoring device through wireless technology or wired lead. The intervention device is selected from one or more of a pressure intervention device, an electric / magnetic stimulation intervention device or a drug intervention device. Although the disclosed attached heart function monitoring and / or intervention system can realize direct local endo / epicardial precise physiological and biochemical function index monitoring and direct local endo / epicardial precise positioning drug delivery or electric / magnetic stimulation or ventricular pressure regulation, and combines monitoring and treatment organically to improve the heart failure state of the patient. However, the obtained biochemical parameters are indirectly obtained by the sensor, and the range and accuracy of the detectable indicators are limited. SUMMARY

[0006] In order to overcome the shortcomings of the prior art, the present application provides a multifunctional heart attachment support device, which provides a basis for multi-method treatment of heart failure patients. The device mainly collects pericardial fluid through the biomimetic suction cup structure on the inner side, and at the same time can realize in-situ drug delivery for the heart. The present application also provides a pericardial fluid collection system and a collection method. The pericardial fluid is collected by the device provided by the present application, and then subjected to biochemical detection. The detectable biochemical indicators include but are not limited to brain natriuretic peptide, cardiac troponin and other common indicators of cardiovascular diseases.

[0007] The technical solution adopted by the present application is:

[0008] In a first aspect, the present application provides a multifunctional heart attachment support device, which is made of elastic photosensitive resin liquid material or soft photosensitive hydrogel material shaped by mold manufacturing or 3D additive manufacturing, and then made by ultraviolet light curing. It should be noted that the mold manufacturing includes "multi-stage shunt bridge" i.e. "ladder type" extrusion of such traditional mold manufacturing methods; for example, Figure 1As shown, the multifunctional heart-attached support device comprises a net structure, an extension column pipe and an extension pipe, the net structure is composed of longitudinally and transversely intersected hollow pipes, and the net structure is sized according to the heart shape of the implantation object, so that the net structure can be loaded and closely attached to the epicardium of the heart surface after being implanted into the chest cavity, to provide physical support for the enlarged heart and limit the adverse expansion of the ventricle; a plurality of micropore suction cups are arranged on the wall of the hollow pipes inside the net structure, the center of the micropore suction cup has a micropore, the micropore is in communication with the hollow pipe to which it is attached, the hollow pipe is in communication with one end of the extension column pipe, the other end of the extension column pipe is used for embedding the extension pipe, and the end of the other end of the extension pipe can be buried subcutaneously and connected with an extracorporeal pipe for collecting pericardial fluid for disease course analysis to provide basis and foundation for precise intervention; the extension pipe can also be connected with a drug pump to realize precise drug delivery at a fixed point, at a fixed time and in a fixed amount through the micropore, and realize real-time mutual intervention of internal and external devices for heart failure.

[0009] The viscosity of the elastic photosensitive resin liquid material is 960-1000 mpa.s, and the physical properties of the elastic photosensitive resin liquid material after being cured by ultraviolet light with a wavelength of 390-420 nm need to meet the following requirements: hardness 70-80D; elastic modulus 1.8-2.4 Mpa; tear strength 40-48.2 KN / M, tensile strength 7-7.9 Mpa, and elongation at break 200-255.1%; preferably, the elastic photosensitive resin liquid material is a white x27 elastic photosensitive resin produced by Polynew Materials Co., Ltd., and the ultraviolet light curing wavelength is 400 nm.

[0010] The soft photosensitive hydrogel material is a mixture of acrylamide and acylurea compounds in an organic solution with a viscosity of 500-800 mpa.s, wherein the mass percentage of acrylamide and acylurea compounds in the total mass of the mixture is 2%-25%, and the implantation device can be printed by a 3D printer under a wavelength of 390-420 nm.

[0011] The soft photosensitive hydrogel material is preferably a mixture of N-acrylamido urea and acrylamide in an organic solution, wherein the mass percentage of N-acrylamido urea and acrylamide in the total mass of the mixture is 10%-25%, and the mass ratio of N-acrylamido urea to acrylamide is 1:(1-1.5).

[0012] After the device is implanted into the chest cavity of a patient, the inner wall suction cup structure forms a micro negative pressure to collect pericardial fluid, the hollow pipelines of the extension column pipe and the net structure are hollow structures and are interconnected, the pericardial fluid flows through the hollow pipelines, is gathered by the hollow pipelines, and then flows through the extension column pipe and the extension pipe in turn, and finally is guided out of the body by the negative pressure for analysis of the disease progression, and subsequent targeted in-situ cardiac drug intervention can be implemented according to the analysis results, the end of the extension pipe is buried subcutaneously after the device is implanted into the body, an external drug pump can be connected at any time, and the treatment drug can directly reach the epicardium to perform in-situ epicardial drug administration in a timed, quantitative and accurate manner. In addition to the pericardial fluid collection and accurate treatment functions, the device can also provide mechanical support for the damaged heart, reduce the ventricular wall stress of the damaged heart, limit the adverse remodeling of the heart, and the hollow design does not cause the problem of pericardial fluid retention.

[0013] Therefore, the device can be used for drug administration treatment of early heart failure patients, and the drug administration is more accurate compared with other drug administration treatment modes, can be used for late heart failure patients to achieve physical support, and can collect epicardial fluid, i.e., pericardial fluid, to analyze the disease progression in a nearly non-invasive manner, and then perform accurate in-situ cardiac drug administration treatment, and the epicardial fluid can be collected again to feedback the treatment effect to form a closed-loop management.

[0014] In the second aspect, the application provides a pericardial fluid collection system, which comprises the multifunctional heart attachment and support device of the first aspect, and further comprises a negative pressure container, an A pipeline, a B pipeline and a peristaltic pump, the subcutaneous leading end of the extension pipe of the device is connected with one end of the A pipeline, the other end of the A pipeline is inserted into the lower space of the negative pressure container, one end of the B pipeline is inserted into the upper space of the negative pressure container, and the other end of the B pipeline is connected with the peristaltic pump.

[0015] In the third aspect, the application further provides a pericardial fluid collection method, which comprises the collection system of the second aspect, and the rotation speed of the peristaltic pump is adjusted to maintain a micro negative pressure environment, when the air pressure in the negative pressure container is stable, the epicardial fluid slowly and uniformly flows out along the A pipeline, and the epicardial fluid is collected, and the negative pressure container needs to be placed in ice water at 0 DEG C during the collection process to avoid degradation of proteins in the sample.

[0016] Compared with the prior art, the application has the following beneficial effects:

[0017] The multifunctional heart-attached support device provided in the first aspect of the present application has a pipeline structure, which is the basis for realizing the multifunction of the device. The device is accommodated in the thoracic cavity to avoid contact with blood and effectively reduce the risk of complications such as thrombosis and infection. The hollow pipelines staggered horizontally and vertically can be implanted subcutaneously through the extension pipes led by the extension columns. When needed, the pipelines at the subcutaneous end can be connected with external instruments. The inner wall is designed with a bionic micropore suction disc structure to provide functions of collecting pericardial fluid and administering drugs in situ. The collected pericardial fluid can be used for disease analysis, and drugs can also be administered in situ according to clinical needs to realize sampling and analysis of pericardial fluid and intelligent and precise administration of drugs in situ.

[0018] The pericardial fluid collection system provided in the second aspect of the present application and the pericardial fluid collection method provided in the third aspect of the present application mainly use the micropore suction disc structure on the inner wall of the multifunctional heart-attached support device to generate a small negative pressure to collect pericardial fluid. Compared with the traditional pericardial fluid collection method, the pericardial fluid can be more accurately collected without mixing with other body fluids in the thoracic cavity. Through in vivo experiments on rats, it is found that, according to the collection results of Example 1 in the specific embodiment of the present application, compared with the device without a suction disc structure, the pericardial fluid collected by the present application is clearer and has a smaller volume without mixing with other body fluids (such as blood). According to Example 2 in the specific embodiment of the present application, proteomics analysis is performed on the pericardial fluid collected from myocardial infarction rats, and many valuable potential therapeutic targets are found. Finally, DNM1L is selected as a drug intervention target, and the cardiac function of myocardial infarction rats is significantly improved after in-situ drug intervention. It is shown that the closed-loop intervention treatment method based on the present application, i.e., "collecting pericardial fluid, analyzing targets / disease, in-situ drug administration, and analyzing feedback of therapeutic effect", is very effective.

[0019] The multifunctional heart-attached support device provided in the present application can take into account early physical auxiliary treatment and late drug administration, and the drug administration is more localized and precise. Meanwhile, the disease development and feedback of therapeutic effect can be monitored through pericardial fluid to form a "monitoring-treatment-feedback" closed loop, so that early intervention and early treatment are realized, and the treatment of heart failure is no longer limited to simple drug treatment and physical constraint treatment. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 FIG. 1 is a schematic diagram of the multifunctional heart-attached support device and its implantation position according to the present application;

[0021] Figure 2 FIG. 2 is a part of the operation photos for manufacturing the multifunctional heart-attached support device in Example 1;

[0022] Figure 3 FIG. 3 is a part of the operation photos for establishing a myocardial infarction model, implanting the device, and collecting pericardial fluid in Example 1; Figure 4Figure 1 is a schematic diagram of the assembly of the pericardial fluid collection system used in Example 1, wherein a is a partial view of the insertion position of the A pipeline and the B pipeline in the negative pressure container; b is a schematic view of the assembly of the pericardial fluid collection system;

[0023] Figure 5 Figure 4 is a schematic diagram of the operation of the epicardial effusion collection using the pericardial fluid collection system in Example 1;

[0024] Figure 6 Figure 5 is a comparison photograph of the epicardial effusion sample collected by the pericardial fluid collection system of the present application and the epicardial effusion sample collected by a conventional device in Example 1, wherein a is the epicardial effusion sample collected by the pericardial fluid collection system of the present application, and b is the epicardial effusion sample collected by a conventional device;

[0025] Figure 7 Figure 6 is a schematic diagram of the micropore structure position of the conventional device without a suction cup structure used for comparison in Example 1;

[0026] Figure 8 Figure 7 is a comparison of the volume of the sample collected by the pericardial fluid collection system of the present application and the sample collected by a conventional device in Example 1 for AMI groups, and a comparison of the volume of the sample collected by the pericardial fluid collection system of the present application for different groups;

[0027] Figure 9 Figure 8 is a schematic diagram of the structure and size of the implant device designed according to the heart of the experimental rat in Example 1. DETAILED DESCRIPTION

[0028] The present application will be further described below in conjunction with the examples and the accompanying drawings.

[0029] Example 1: Collection of pericardial fluid of acute myocardial infarction rats using the pericardial fluid collection system of the present application

[0030] 1.1 Experimental materials

[0031] Experimental animals: 30 male Sprague-Dawley (SD) rats (250±20g) were selected for this experiment. All animals were purchased from Hangzhou Medical College, quality license number: SCXK (Zhejiang) 2019-0002, and qualified certificate number: 20210712Aazz0100000252. All animal experiments followed the guidelines and ethical guidelines for animal experiments of China Pharmaceutical University, and conformed to the Helsinki Declaration.

[0032] Experimental reagents: Flexible resin was purchased from Poly Xinxin Materials; ultrapure water was purchased from Fisher Chemical; sodium chloride was purchased from Nanjing Chemical Reagent Co., Ltd.; chloral hydrate was purchased from Sinopharm Chemical Reagent Co., Ltd.; anhydrous ethanol was purchased from Nanjing Chemical Reagent Co., Ltd.; disodium hydrogen phosphate dodecahydrate was purchased from Nanjing Chemical Reagent Co., Ltd.; sodium hydroxide was purchased from Xilong Science Co., Ltd.; sodium bicarbonate was purchased from Nanjing Chemical Reagent Co., Ltd.; potassium dihydrogen phosphate was purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd.; potassium chloride was purchased from Shanghai Lingfeng Chemical Reagent Co., Ltd.; iodine disinfectant was purchased from Shandong Anjie High-tech Disinfection Technology Co., Ltd.; and dipotassium hydrogen phosphate was purchased from Nanjing Chemical Reagent Co., Ltd.

[0033] Experimental instruments: Animal physiological function experimental system BL-420S was purchased from Chengdu Taimeng Co., Ltd.; small animal ventilator HX-300S was purchased from Chengdu Taimeng Co., Ltd.; 1 / 10,000 electronic balance AL204 was purchased from Mettler-Toledo Instrument Co., Ltd.; pressure converter YP100 was purchased from Beijing Xinhang Xingye Science and Technology Trade Co., Ltd.; pure water working instrument SMART-N was purchased from Shanghai Kanglei Analytical Instrument Co., Ltd.

[0034] 1.2 Experimental methods

[0035] Device structure and size design: Figure 9 As shown, the specific parameters of the device in this embodiment are as follows: The device's various parameters were determined by measuring the major and minor axes of rat hearts. The main body of the two top extension tubes of the device has a diameter of 2 mm and a length of 10 mm. The largest circle in the main body of the device has an inner diameter of 13 mm and an outer diameter of 15 mm; the smallest circle has an inner diameter of 3 mm and an outer diameter of 5 mm. Furthermore, the hollow tube has an inner diameter of 0.6 mm and a wall thickness of 0.4 mm. Microporous suction cups are evenly distributed on the inner wall. The overall diameter of the microporous suction cups is 1.2 mm, and the micropores of the microporous suction cups have a pore diameter of 300 μm.

[0036] Fabrication of implantable devices: Figure 2 As shown, first, Figure 2 As shown in Figure a, use 3D printing software to slice the electronic 3D drawings of this device and convert them into 3D printing format; Figure 2The elastic liquid photosensitive resin X23 is selected as the elastic liquid photosensitive resin, as shown in the b diagram in the figure, and the viscosity is 980 mpa.s. 100-200 ml of the resin is added to the material tray of the 3D printer. The printer parameter settings are as follows: light curing wavelength 395-405 nm, precision (i.e. layer thickness) 0.05 mm, layer number 15-25, bottom layer exposure time 20-25 s, conventional exposure time 1.7-3.5 s, adhesion molding time 30.0 s, shell wall thickness 2.0 mm, and filling wall thickness 1.0 mm. The curing time of each layer is 0.2 s. The 3D printer of model Proxima 6.0 is used for printing, and the total printing time is about 1 h 30 min. After printing, the model is taken out, ultrasonic cleaning in anhydrous ethanol for 10 min, the support column is cut off, and the finished product of the microporous suction cup device is obtained by secondary curing under a wavelength of about 400 nm for 5 min, as shown in the c diagram in the figure. Figure 2 As shown in the d, e and f diagrams in the figure, the weight of the finished product of the same batch is 0.7-0.8 g. The finished product sample is subjected to mechanical property testing, and the hardness is 75D, the elastic modulus is 2.1 Mpa, the tear strength is 48.2 KN / M, the tensile strength is 7.9 Mpa, and the elongation at break is 255.1%. In order to reflect the collection effect of the microporous suction cup of the multifunctional heart attachment support device provided in the present application, another conventional net cover device without a biomimetic suction cup and only with micropores is made according to the device size, material and manufacturing process of the present application, Figure 7 As shown in the figure, the difference between the conventional net cover device for comparison and the multifunctional heart attachment support device provided in the present application is that the conventional device is directly provided with a plurality of micropores on the hollow pipeline wall inside the net cover structure, without a suction cup structure.

[0037] Animal grouping: 30 male Sprague-Dawley (SD) rats (250±20 g) were selected for the experiment. All animals were purchased from Hangzhou Medical College, with quality license number SCXK (Zhejiang) 2019-0002 and qualified certificate number 20210712Aazz0100000252. All animal experiments followed the animal experiment guidelines and animal experiment ethical guidelines of China Pharmaceutical University, and met the Helsinki Declaration. They were divided into acute myocardial infarction group (AMI) and sham operation group (Control). The acute myocardial infarction group (AMI) was established by ligating the left anterior descending branch of the coronary artery to establish a myocardial infarction model; the sham operation group (Control) was established by passing a needle and thread through the ligation point after thoracotomy without ligation.

[0038] The process of establishing an acute myocardial infarction (AMI) model is as follows: all rats were weighed; if Figure 3 As shown in the a diagram in the figure, 10% chloral hydrate was used to anesthetize the rats at a dose of 0.3 g / kg by intraperitoneal injection; after the rats were anesthetized, the chest was opened by invasive surgery, as shown in the b diagram in the figure. Figure 3b, c and d of FIG. 1, after fixing the rat in supine position, the skin is cut at the position 1.5-2.0 cm above the xiphoid process of the rat and 2-3 mm to the right with ophthalmic scissors; if Figure 3 e and f of FIG. 1, after separating the skin with hemostatic forceps, the pectoral muscle is seen on the anterior serratus muscle, the pectoral muscle is first held and separated bluntly to separate it from the anterior serratus muscle, then the pectoral muscle is held up, the anterior serratus muscle is seen on the rib, the anterior serratus muscle is held up and separated bluntly to separate it from the rib, the separated muscle and skin are spread with the speculum to expose the intercostal space of the third and fourth ribs to form the heart surgery field; if Figure 3 g of FIG. 1, at this time, the pericardium on the surface of the heart is gently torn with forceps to find the position of the left coronary vein of the heart, and a 6.0 suture is used to ligate the left anterior descending branch (LAD) of the coronary artery 1-2 mm below the tip of the left auricle. The occurrence of acute myocardial infarction is confirmed by color change in the ischemic area of the heart and characteristic changes in electrocardiogram, and the acute myocardial infarction model rat surgery group, i.e. the AMI group, is divided into two subgroups, one of which is used to implant the multifunctional heart attaching and supporting device designed and made according to the present application, and the other of which is used to implant the conventional device; another group of sham operation groups is set, the sham operation group does not ligate the left anterior descending branch of the coronary artery but only threads, and the rest of the operation is the same as before, and the same sham operation group is also divided into two subgroups, one of which is used to implant the multifunctional heart attaching and supporting device designed and made according to the present application, and the other of which is used to implant the conventional device.

[0039] Device implantation: according to the above grouping, the device designed and made according to the present application and the conventional device used for comparison are implanted into the corresponding rats according to the grouping; it should be noted that, because the experimental object is a rat, the individual is small, and in order to simplify the experimental materials and experimental operation, in the present experimental example, the A pipeline serves as an extension pipe; invasive minimally invasive thoracotomy is used for implantation, the sterilized device is folded and sent into the apex of the rat heart, after reaching the apex, the device is unfolded, the extension column pipe of the device is held with forceps, and the device is sleeved on the extension column pipe from the apex to the bottom of the heart; the process must be gentle to avoid damaging other organs; if Figure 3 h, i and j of FIG. 1, the extension column pipe of the device is threaded out from the second and third intercostal spaces of the rat, one end of the A pipeline is sleeved into the hollow extension column pipe with forceps, and then the A pipeline is led subcutaneously from the forelimb to the neck back subcutaneously and threaded out, connected with the negative pressure container; if Figure 3 k and l of FIG. 1, after all the operations are completed, the muscle layers are sutured layer by layer, and at the same time, the air in the thoracic cavity of the rat is drawn back to form a negative pressure in the thoracic cavity to help the rat recover spontaneous breathing, and the rat is placed on a heating plate after the operation to keep the body temperature of the rat to prevent postoperative hypothermia.

[0040] Collection of pericardial fluid: if Figure 4As shown, the subcutaneously introduced in-vitro pipeline of the above-mentioned four groups of rats with device implantation is connected with the A pipeline of the negative pressure container, the B pipeline of the negative pressure container is connected with the peristaltic pump, the rotation speed of the peristaltic pump is adjusted to maintain a micro-negative pressure environment, when the air pressure in the negative pressure container is stable, the epicardial exudate in the A pipeline slowly and uniformly flows out, and the epicardial exudate is collected. As shown in Figure 5 As shown, this process will always immerse the negative pressure container in ice water (0℃) to avoid protein degradation in the sample. After continuous collection for 6 hours, the sample is transferred to a centrifuge tube, centrifuged at 2000xg for 10 min at 4℃, and the supernatant is taken after measuring the sample volume and stored at -80℃.

[0041] 1.3. Experimental results

[0042] The pericardial fluid collected in the above-mentioned four groups of rats is statistically analyzed and analyzed, as shown in Figure 6 As shown in a graph in the figure, the multifunctional heart attaching and supporting device with microporous suction cups of the application can successfully collect epicardial exudate, and the naked eye observation is a light yellow clear liquid without visible impurities and blood, as shown in Figure 6 b graph in the figure is the epicardial exudate collected by the conventional device without suction cups, which is relatively turbid in appearance, and the color is reddish and yellow. As shown in Figure 8 As shown in the figure, the statistical analysis results of the volume of epicardial exudate collected by the multifunctional heart attaching and supporting device with microporous suction cups of the application show that the sample volume of the control group is 0.97±0.17mL, and the sample volume of the AMI group is 1.25±0.16mL. Compared with the blank group, the volume of epicardial exudate in the AMI group is significantly more than that in the sham operation group, with a very significant difference (P<0.01); the statistical analysis of the sample volume of the same group by the conventional device and the microporous suction cup device shows that compared with the conventional device, the sample volume collected by the microporous suction cup device in the blank group and the AMI group is significantly less than that of the conventional device (P<0.01), which has statistical significance. It shows that the multifunctional heart attaching and supporting device with microporous suction cups of the application has higher specificity for epicardial exudate, which avoids the simultaneous collection of blood and chest cavity exudate and other organ surface exudate with epicardial exudate.

[0043] Proteomic analysis and screening of differentially expressed proteins (DEPs) in pericardial fluid after myocardial infarction: Non-labeled proteomic analysis was performed on epicardial exudate collected by the conventional device described above, and a total of 651 proteins were identified, of which 140 were up-regulated and 98 were down-regulated. TMT-labeled proteomic analysis was performed on epicardial exudate collected by the multifunctional heart-attached support device with microporous suction disc according to the application, and a total of 1292 proteins were identified, of which 77 were up-regulated and 49 were down-regulated. Through protein-protein interaction network (PPI) analysis, it was found that ubiquitin-60S ribosomal protein L40 (Uba52), eukaryotic elongation factor 2 kinase (Eef2k), dynamin-related protein 1 (DNM1L), heat shock homologous protein 71 kDa (Hspa8), 60S ribosomal protein L4 (Rpl4), eukaryotic translation initiation factor 3 subunit I (Eif3i), integrin alpha M (Itgam), 60S ribosomal protein L13a (Rpl13a), nucleoside diphosphate kinase B (Nme2), protein disulfide isomerase A3 (Pdia3), 14-3-3 protein gamma (Ywhag), protein phosphatase 2 (Ppp2r1a), etc. Proteins are in the key nodes in the PPI network. They are mainly involved in myocardial contraction, protein synthesis regulation, complement and inflammatory response processes, etc.

[0044] In summary, the microporous suction disc biomimetic device improves the specificity of pericardial fluid collection, making the proteomic results more accurate and reliable. Laboratory proteomic analysis of epicardial exudate in the early stage after acute myocardial infarction found that most of the differentially expressed proteins in the pericardium have complex roles in the early stage of AMI and the compensatory adaptation process after ischemia, which may be potential target points for regulating epicardium to promote cardiac repair.

[0045] Example 2: In situ drug intervention in rats with myocardial infarction based on proteomic target points collected from pericardial fluid

[0046] In order to explore new drug treatment targets for myocardial infarction, by screening the top 5% hub proteins in the proteomic results of pericardial fluid, DNM1L was finally determined as a potential therapeutic target for myocardial infarction (heart attack). This example uses DNM1L protein inhibitor Mdivi1 to prove the scientificity of the analysis target point by in situ drug administration on the epicardium of rats with myocardial infarction by the device.

[0047] 2.1 Experimental materials

[0048] Experimental animals: 18 male Sprague-Dawley (SD) rats (250 ± 20 g) were selected for this experiment. All animals were purchased from Hangzhou Medical College, quality license number: SCXK (Zhejiang) 2019-0002. All animal experiments comply with the guidelines and ethical guidelines for animal experiments of China Pharmaceutical University, and comply with the Helsinki Declaration.

[0049] Experimental reagents: Sodium chloride was purchased from Nanjing Chemical Reagent Co., Ltd.; Chloral hydrate was purchased from Sinopharm Chemical Reagent Co., Ltd.; Anhydrous ethanol was purchased from Nanjing Chemical Reagent Co., Ltd.; Disodium hydrogen phosphate dodecahydrate was purchased from Nanjing Chemical Reagent Co., Ltd.; Sodium hydroxide was purchased from Xilong Scientific Co., Ltd.; Sodium bicarbonate was purchased from Nanjing Chemical Reagent Co., Ltd.; Potassium dihydrogen phosphate was purchased from Shanghai Lingfeng Chemical Co., Ltd.; Potassium chloride was purchased from Shanghai Lingfeng Chemical Co., Ltd.; Neutral gum sealing agent was purchased from Shanghai Sangon Biological Engineering Co., Ltd.; Xylene was purchased from Nanjing Chemical Reagent Co., Ltd.; Glacial acetic acid was purchased from Nanjing Chemical Reagent Co., Ltd.; Concentrated hydrochloric acid was purchased from Nanjing Chemical Reagent Co., Ltd.; Eosin was purchased from Beijing Chemical Reagent Company; Hematoxylin was purchased from Beijing Chemical Reagent Company; Iodophor disinfectant was purchased from Shandong Anjie Gaoke Disinfection Technology Co., Ltd.; Heparin sodium was purchased from Beijing Solabio Technology Co., Ltd.; TM-100 medical coupler was purchased from Tianjin Jin Ya Science and Technology Development Co., Ltd.; Disodium hydrogen phosphate was purchased from Nanjing Chemical Reagent Co., Ltd.

[0050] Experimental instruments: Animal physiological function experiment system BL-420S was purchased from Chengdu Taimeng Co., Ltd.; Small animal respirator HX-300S was purchased from Chengdu Taimeng Co., Ltd.; One-hundredth electronic balance AL204 was purchased from Mettler-Toledo Instrument Co., Ltd.; Pressure converter YP100 was purchased from Beijing Xinhangxing Industry and Trade Co., Ltd.; Pure water working instrument SMART-N was purchased from Shanghai Kanglei Analysis Instrument Co., Ltd.; Small animal ultrasonic imaging system was purchased from Baosheng Medical Equipment Co., Ltd.; Electric oven XMTD-8222 was purchased from Shanghai Jinghong Experimental Equipment Co., Ltd.

[0051] 2.2 Experimental method

[0052] Animal grouping: 18 SD rats (200-220 g, male) were randomly divided into 3 groups: sham operation group (Control group), acute myocardial infarction group (AMI), and heart infarction treatment group (Treatment); Control group was a sham operation group, after thoracotomy only threading without ligation; AMI group was performed myocardial infarction model establishment, without any treatment; Treatment group implanted micro-pore suction disc device after modeling, within 1 day after coronary artery ligation, injected mdivi-1 in situ through the extension tube, the dose was 1.2 mg / Kg.

[0053] Disease model establishment: The acute myocardial infarction (AMI) model establishment process was as described in Example 1.

[0054] Methods and indexes for evaluating heart function include echocardiography and hemodynamics. Echocardiography detects indexes such as left ventricular ejection fraction (LVEF%), left ventricular fractional shortening (LVFS%), left ventricular internal dimension at systole (LVIDs) and left ventricular internal dimension at diastole (LVIDd), which are important indexes for evaluating myocardial infarction. Hemodynamics obtains values such as left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximum ascending rate of left ventricular internal pressure (+dp / dtmax) and maximum descending rate of left ventricular internal pressure (-dp / dtmax).

[0055] Implantation of the device: The device implantation procedure was as described in Example 1.

[0056] 2.3 Experimental results

[0057] The echocardiography results are shown in Table 1 below, in which the data of each group is represented by mean ± standard deviation (Mean ± SD). (*** represents P<0.001 compared with the sham operation group; #, ## represent P<0.05 and P<0.01, respectively, compared with the acute myocardial infarction group.

[0058] Table 1: Values of left ventricular ejection fraction (LVEF%), left ventricular fractional shortening (LVFS%), left ventricular internal dimension at systole (LVIDs) and left ventricular internal dimension at diastole (LIVDd) of rats in each group on day 28

[0059]

[0060] The data in Table 1 above show that, compared with the acute myocardial infarction group, the values of left ventricular ejection fraction (LVEF%) and left ventricular fractional shortening (LVFS%) of the myocardial infarction rats were significantly increased after treatment with Mdivi1 by administration of the device.

[0061] The hemodynamics results are shown in Table 2 below, in which the data of each group is represented by mean ± standard deviation (Mean ± SD); ** represents P<0.01 compared with the Control group.

[0062] Table 2: Values of left ventricular systolic pressure (LVSP), left ventricular end-diastolic pressure (LVEDP), maximum ascending rate of left ventricular internal pressure and maximum descending rate of left ventricular internal pressure (+dp / dtmax) of rats in each group on day 28

[0063]

[0064] As shown in the above table, the left ventricular internal pressure maximum rising rate (+dp / dtmax) value of the treatment group is higher than that of the AMI group.

[0065] In summary, the experiment of Example 2 evaluates the heart function of rats in each experimental group by echocardiography and hemodynamic data. The experimental results show that, based on the pericardial fluid analysis of the target, the corresponding intervention drug given by the device can effectively improve the left ventricular systolic pressure (LVSP), the left ventricular fractional shortening (LVFS%), and the left ventricular internal pressure maximum rising rate (+dp / dtmax) value of the rats after myocardial infarction, and improve the heart function of the myocardial infarction rats. Example 2 proves the ingenuity and feasibility of the design of the present application, and verifies the closed-loop treatment concept of "collecting body fluid-analyzing target / disease-precise intervention" based on the present application, and preliminarily embodies the application prospect of the device.

Claims

1. A multifunctional cardiac support device, characterized by: The multifunctional heart attachment support device is made of elastic photosensitive resin liquid material or soft photosensitive hydrogel material through mold manufacturing or 3D additive manufacturing, and then cured by ultraviolet light. The multifunctional heart attachment support device includes a mesh structure, an extension column tube and an extension tube. The mesh structure is composed of crisscrossing hollow tubes, and the mesh structure is customized in size according to the shape of the heart of the implanted object, so that the mesh structure can be wrapped and tightly attached to the epicardium on the surface of the heart after implantation in the chest cavity, providing physical support for the enlarged heart and limiting the adverse expansion of the ventricle; a plurality of microporous suction cups are provided on the wall of the hollow tube inside the mesh structure, and the center of the microporous suction cup has a micropore, and the micropore is connected to the hollow tube to which it is attached, and the hollow tube is connected to one end of the extension column tube, and the other end of the extension column tube is used to nest the extension tube. The end of the other end of the extension tube can be buried subcutaneously and can be connected to an extracorporeal tube; the microporous suction cup is used to collect pericardial fluid; The viscosity of the elastic photosensitive resin liquid material is in the range of 960-1000 mPa.s. The physical properties of the elastic photosensitive resin liquid material after being cured by ultraviolet light with a wavelength of 390-420 nm must meet the following requirements: hardness 70-80D; elastic modulus 1.8-2.4 MPa; tear strength 40-48.2 KN / M, tensile strength 7-7.9 MPa, and elongation at break 200-255.1%. The soft photosensitive hydrogel material is a mixture of acrylamide and urea compounds in an organic solution with a viscosity of 500-800 mPa.s, wherein the acrylamide and urea compounds each account for 2%-25% of the total mass of the mixture. The multifunctional cardiac attachment support device can be printed using a 3D printer at a wavelength of 390-420 nm.

2. The multifunctional cardiac support device according to claim 1, characterized in that: The physical properties of the elastic photosensitive resin liquid material after being cured by ultraviolet light with a wavelength of 400nm meet the following requirements: hardness 75D, elastic modulus 2.1Mpa, tear strength 48.2KN / M, tensile strength 7.9Mpa, elongation at break 255.1%, and the ultraviolet curing wavelength is 400nm.

3. The multifunctional cardiac support device according to claim 1, characterized in that: The soft photosensitive hydrogel material is a mixture of N-acryloyl semicarbazide and acrylamide in an organic solution, wherein the percentage of N-acryloyl semicarbazide and acrylamide in the total mass of the mixture is 10% to 25%, and the mass ratio of N-acryloyl semicarbazide to acrylamide is 1:(1 to 1.5).

4. A pericardial fluid collection system, characterized by: The system includes a multifunctional cardiac attachment support device as described in claim 1, and also includes a negative pressure container, pipe A, pipe B and a peristaltic pump. The subcutaneous lead-out end of the extension column tube of the multifunctional cardiac attachment support device is connected to one end of the pipe A, the other end of the pipe A is inserted into the lower space of the negative pressure container, one end of the pipe B is inserted into the upper space of the negative pressure container, and the other end of the pipe B is connected to the peristaltic pump.

5. The pericardial fluid collection system according to claim 4, characterized in that: The pericardial fluid collection system is used to adjust the peristaltic pump speed to maintain a slightly negative pressure environment. When the air pressure in the negative pressure container is stable, the epicardial exudate flows out slowly and evenly along the A channel, and the epicardial exudate can be collected. During the collection process, the negative pressure container needs to be placed in ice water at 0°C to avoid protein degradation in the sample.

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