CN104130437B - A hard brain / meninges biological patch material and its preparation method and application - Google Patents

A hard brain / meninges biological patch material and its preparation method and application Download PDF

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CN104130437B
CN104130437B CN201410373258.6A CN201410373258A CN104130437B CN 104130437 B CN104130437 B CN 104130437B CN 201410373258 A CN201410373258 A CN 201410373258A CN 104130437 B CN104130437 B CN 104130437B
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meninges
material
biological
dura
soaking
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CN104130437A (en
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孙先昌
徐秀
董佳桓
郭松
郭刚
王彬
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苏州正海生物技术有限公司
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Abstract

本发明提供了一种硬脑/脊膜生物补片材料及其制备方法和应用。 The present invention provides a hard brain / meninges biological patch material and its preparation method and application. 所获得的生物补片材料特别适用于硬脑/脊膜缺损的修复,在植入人体或动物体内后,能够在体内保持6月‑2年时间内稳定不降解,当超过2年后会逐渐降解,保证具有一定力学性能以及防液体渗漏作用的同时还具有生物相容性,产生术后并发症以及副作用的风险大大降低,满足了对于用于修复硬脑/脊膜缺损材料的特殊要求。 The obtained biological patch material is particularly suitable for the repair of hard brain / meninges defect, after implantation in the human or animal body, can be held in June in vivo - a stable non-degradable within 2 years, after more than 2 years will gradually degradation, while ensuring a certain mechanical properties as well as the liquid leakage prevention effect is also biocompatible, the risk of complications, and greatly reduces side effects, to meet the special requirements for repairing dura / mater defect material .

Description

一种硬脑/脊膜生物补片材料及其制备方法和应用 A hard brain / meninges biological patch material and its preparation method and application

技术领域 FIELD

[0001] 本发明属于组织工程学医用生物材料技术领域,具体涉及一种硬脑/脊膜生物补片材料及其制备方法和应用。 [0001] The present invention belongs to the technical field of tissue engineering medical biomaterials, particularly relates to a hard brain / meninges biological patch material and its preparation method and application.

背景技术 Background technique

[0002] 硬脑膜和硬脊膜分别是大脑与颅骨、脊柱与脊髓之间重要的功能膜组织,具有相同的生物学特征和功能。 [0002] The dura mater and the dura are important functional film tissue between the skull and the brain, spine and spinal cord, it has the same biological features and functions. 硬脑膜贴附颅腔内面,与颅盖连结疏松,与颅底结合紧密;硬脑膜在脑神经颅处向外延伸,移行而与神经的被膜结合,在枕骨大孔周缘向下延为硬脊膜。 Attaching the inner surface of the cranial dura, with loose coupling calvaria, in close conjunction with the base of the skull; brain cranial dura extends outwardly at the transitional coating and Neural in the large hole rim extending downward occipital dura . 硬脑膜与硬脊膜的功能和结构是一致的,硬脑膜用于保护脑组织,硬脊膜用于保护脊髓组织。 Dura and dural function and structure is the same, to protect the brain dura mater, epidural spinal cord tissue for protection.

[0003] 外伤、肿瘤、炎症、神经外科手术或其他脑颅脊柱疾病等均会造成硬脑/脊膜缺损。 [0003] trauma, cancer, inflammation, or other cranial neurosurgery spine disease etc. will cause hard brain / meninges defect. 硬脑/脊膜的缺损会引起癫痫、脊液外漏、颅内感染、脑膜膨出和脑功能障碍等病症。 Dura / mater defect can cause seizures, spinal fluid leak, intracranial infections, meninges and brain dysfunction and other diseases. 如在颅底外科手术中,颅底硬脑膜相对薄且与颅底骨结合紧密,一旦出现缺损易造成脑脊液鼻漏和耳漏。 As in skull base surgery, the skull base dura relatively thin and closely combined with the skull base, once the defect occurs easily causing leakage of cerebrospinal fluid and otorrhea. 颅底的脑膜瘤、脊索瘤等肿瘤常常侵蚀浸润到周围的颅骨和硬脑膜,在切除肿瘤手术时,切除的肿瘤通常也会附带部分脑膜和颅骨,造成术后发生脑脊液漏和颅内感染。 Skull base meningioma, chordoma and other tumors are often aggressive infiltration into the skull and dura around in the surgical removal of the tumor, resection of the tumor will usually incidental part of the meninges and skull, resulting in postoperative leakage of cerebrospinal fluid and intracranial infection.

[0004] 为治疗硬脑/脊膜缺损,现有技术中常采用硬脑/脊膜替代材料植入缺损部分进行修复硬脑/脊膜缺损。 [0004] for the treatment of hard brain / meninges defect, the prior art often used Dura / mater substitute material implanted to repair the defect portion of hard brain / meninges defect. 随着医学技术的高速发展,硬脑膜替代材料越来越广泛的应用于脑凸面手术、创伤直接损伤、肿瘤的浸润、手术过程中硬脑膜切开减压以及一些先天性的因素造成的硬脑膜缺损等诸多方面,如在脑外科开颅手术患者中有10%〜15%需要硬脑/脊膜替代材料修复硬脑膜缺损,也有报道称该比例已经高达30%。 With the rapid development of medical technology, dura substitute materials more and more widely used in brain surgery convex, direct trauma injury, tumor invasion, dural incision during surgery as well as some congenital dural decompression caused by factors defects and other aspects, as there are in brain surgery craniotomy 10% ~ 15% of patients in need dura / repair of the dura mater substitute material defect, also reported that the ratio has been as high as 30%. 因此,为了满足临床需要,开展研究合适的硬脑/脊膜修复材料至关重要。 Therefore, in order to meet clinical needs, research the appropriate hard brain / meninges vital repair material.

[0005] 但是,现有的硬脑/脊膜替代材料在使用的过程中往往存在诸多不足,常常引起严重的并发症问题。 [0005] However, the existing hard brain / meninges alternative materials used in the process often has many deficiencies, problems often cause serious complications. 例如在硬脑/脊膜替代材料在植入体内后降解速度控制方面的问题,若硬脑/脊膜替代材料降解的速度过快,撕裂力下降,则导致防液体渗漏作用下降,引起硬脑/脊膜与周围组织粘连、感染、出血、脑脊液漏以及癫痫等病症;但是若硬脑/脊膜替代材料长时间不降解,则又导致其与组织再生不匹配,生物相容性差,更甚者易导致炎症反应、结缔组织增生包裹、感染及出血等问题。 For example, in the dura / mater substitute material after implantation in vivo degradation rate control issues, if the dura / mater substitute material too fast degradation, tear strength decreased, resulting in the effect of preventing leakage of liquid drops, causing hard brain / meninges surrounding tissue adhesion, infection, hemorrhage, cerebrospinal fluid leakage and disorders such as epilepsy; However, when the dura / mater substitute material does not degrade for a long time, which in turn results in a mismatch and tissue regeneration, biocompatible difference, What is more easily lead to inflammation, connective tissue proliferation parcels, infection and bleeding problems. 因此,对于硬脑/脊膜替代材料在植入人体或动物体内的降解速度方面的控制是非常重要的。 Thus, the material for dura mater substitute control / implantation in the human or animal body degradation rate is very important.

[0006] 然而,在以往以天然膜材料生产硬脑/脊膜替代材料的工艺中,当膜材料经过冻干辐照后,天然膜材料中的胶原蛋白会被大幅破坏,使膜材料撕裂力下降,以及为了能够更好脱去细胞和脂肪,使用的脱细胞、抗原和脂肪的试剂对膜的破坏相对较大,使天然膜材料的其他力学性能大幅降低,最终导致其降解速度加快,防液体渗漏作用下降。 [0006] However, in the conventional natural membrane materials production process dura / meningeal substitute material, the film material when irradiated after lyophilization, natural collagen membrane material is substantially destroyed, tearing of the film material power drops, and in order to better removal of fat cells and acellular use, antigens and reagents fat damage to the membrane is relatively large, so that the other mechanical properties of natural film material greatly declined, eventually leading to its degradation speed, liquid leakage prevention effect decreases. 为了降低生物组织修复材料的降解速度,现有技术中通常采用交联剂处理膜材料,提高其力学性能,降低其降解速度,使其在人体或动物体内能够长时间不降解。 To reduce the rate of degradation of biological tissue repair materials, the prior art commonly employed cross-linking agent in the film material, improve the mechanical properties, reducing the degradation rate, it is possible to not degrade for a long time in the human or animal body. 但是对于用于修复硬脑/脊膜缺损的替代材料来说,由于其特殊的生理位置以及功能决定了现有技术中用于处理其他部位的组织修复材料或硬脑/脊膜缺损的替代材料中的交联剂并不适用于硬脑/脊膜补片材料,主要存在的问题包括:(1)交联剂处理后,修复材料在人体或动物体内长时间不降解,导致其在植入人体或动物体内后,生物相容性差,易产生不利的副作用;(2)交联剂的稳定性低,大多数交联剂都具有比较容易反应的基团,这种基团导致交联剂不稳定,进而导致修复材料在人体或动物体内短时间内即降解,导致修复材料防液体渗漏作用下降,进而引发一系列术后并发症,带来不利的影响;(3)交联剂毒性高,难以控制交联剂最终在产品上的残留量,影响被植入的人或动物的身体健康。 However, for repairing hard for brain / meninges defect alternative materials, because of its special physiological functions and determines the position of the prior art tissue repair material for processing or other parts of the dura substitute material / defect meninges the crosslinking agent is not suitable for hard brain / meninges patch material, the main problems include: (1) cross-linking agent, restorative material does not degrade for a long time in the human or animal body, which implant leads after the human or animal body, poor biocompatibility, easy to produce adverse side effects; low stability (2) a crosslinking agent, a crosslinking agent having a relatively easy most reactive groups, such groups cause a crosslinking agent instability, which led to the repair material that is degraded in the human or animal body a short time, resulting in leakage of liquid anti-repair material effect decline, triggering a series of complications and adverse effects; (3) a crosslinking agent toxicity high, it is difficult to control the amount of crosslinking agent remaining in the final product, affecting the health of implanted human or animal. 因此,现有技术中亟需开发一种更为适用于硬脑/脊膜修补降解性能要求的生物补片材料。 Thus, the prior art need for the development of a more suitable dura / meningeal repair patch materials of biological degradation of performance requirements.

发明内容 SUMMARY

[0007] 为此,本发明所要解决的技术问题在于现有技术中硬脑/脊膜替代材料植入人体后降解速度过快或过慢均不利于修复硬脑/脊膜缺损的问题,进而提供一种适用于硬脑/脊膜缺损修复降解要求的硬脑/脊膜生物补片材料,并公开其制备方法和应用。 [0007] To this end, the present invention is to solve the technical problem of the prior art dura / mater substitute material implanted too fast or too slow degradation are not conducive to repair dura / mater defect issues, and further to provide a suitable hard brain / meninges degradation defect claim hard brain / meninges biological patch material is disclosed and their preparation and use.

[0008] 为解决上述技术问题,本发明提供了一种硬脑/脊膜生物补片材料的制备方法,包括取动物膜组织为原材料,并顺次置于表面活性剂溶液、碱液、过氧化物溶液、质量浓度为l-5wt%的缩水甘油醚类环氧交联剂溶液以及磷酸盐缓冲液中进行浸泡的步骤,并将经上述各步骤浸泡后的所述膜组织材料冷冻干燥后经辐照灭菌,即得。 [0008] In order to solve the above technical problem, the present invention provides a method for preparing a hard brain / meninges biological patch material, including a film take animal tissues as raw material, and sequentially placed in a surfactant solution, lye, too after soaking step glycidyl ether epoxy crosslinking oxide solution and phosphate buffer solution, the concentration of l-5wt%, the tissue material and freezing the film through the respective steps of drying after immersion by irradiation sterilization, that is.

[0009] 所述的硬脑/脊膜生物补片材料的制备方法,所述缩水甘油醚类环氧交联剂溶液浸泡的步骤中,所述交联剂为丙三醇缩水甘油醚和/或乙二醇缩水甘油醚。 [0009] The hard brain / meninges method for preparing a biological patch material, a glycidyl ether epoxide step of soaking in a solution of crosslinking agent, the crosslinking agent is a glycerol ether and / or glycol ether.

[0010] 所述的硬脑/脊膜生物补片材料的制备方法,所述缩水甘油醚类环氧交联剂溶液浸泡的步骤中,所述浸泡时间为15h_lOd。 [0010] The method of preparing the hard brain / meninges biological patch material, said step of glycidyl ether epoxide crosslinking agent solution soaking, the soaking time is 15h_lOd.

[0011] 优选的,所述膜组织材料在浓度为2_4wt%的交联剂中浸泡20h。 [0011] Preferably, the membrane material is soaked tissue 20h 2_4wt% at a concentration of crosslinking agent.

[0012] 更优选的,所述膜组织材料在浓度为2wt%的交联剂中浸泡。 [0012] More preferably, the material is soaked in the membrane tissue concentration of 2wt% of crosslinking agent.

[0013] 所述的硬脑/脊膜生物补片材料的制备方法,所述含表面活性剂溶液浸泡的步骤中,所述表面活性剂的质量浓度为〇. l_5wt%,并控制所述溶液温度为0_25°C进行浸泡。 [0013] The hard brain / meninges method for preparing a biological patch material, the step of soaking the surfactant solution containing, the mass concentration of the surface active agent is square. L_5wt%, and controls the solution soaking temperature to 0_25 ° C.

[0014] 所述的硬脑/脊膜生物补片材料的制备方法,所述碱液浸泡的步骤中,所述碱液的浓度为〇. I-5mo 1/L,并控制所述溶液温度为5-15 °C进行浸泡。 [0014] The method of preparing the hard brain / meninges biological patch material, the step of soaking in alkali solution, the alkali concentration is square. I-5mo 1 / L, and controlling the temperature of the solution soaking is 5-15 ° C.

[0015] 优选的,在所述碱液浸泡的步骤中,膜组织材料加入至浓度为lmol/L的氢氧化钠溶液中浸泡2h,并控制溶液温度为8 °C。 [0015] Preferably, in the step of alkali soaking, the membrane tissue material is added to a concentration of lmol / L sodium hydroxide solution soaking 2h, and controlling the temperature of the solution is 8 ° C.

[0016] 所述的硬脑/脊膜生物补片材料的制备方法,所述过氧化物溶液浸泡的步骤中,所述过氧化物溶液的质量浓度为l_5wt%。 Preparation dura [0016] the / meninges biological patch material, the peroxide solution soaking step, the concentration of the peroxide solution was l_5wt%.

[0017] 优选的,在所述过氧化物溶液浸泡的步骤中,浸泡后的膜组织材料加入至浓度为2wt %过氧化物溶液中浸泡处理I -5h。 [0017] Preferably, in the step of immersion of the peroxide solution, the membrane material was soaked tissue was added to a concentration of 2wt% peroxide solution soaking I -5h.

[0018] 优选的,所述过氧化物溶液为过氧化氢溶液。 [0018] Preferably, the peroxide solution is a hydrogen peroxide solution.

[0019] 所述的硬脑/脊膜生物补片材料的制备方法,所述辐照灭菌的步骤为电子束辐照灭菌,辐照计量为10_30KGy。 Preparation dura [0019] the / meninges patch biological material, the radiation sterilization step is electron beam radiation sterilization, radiation measurement of 10_30KGy.

[0020] 优选的,所述电子束的辐照计量为25KGy。 [0020] Preferably, the irradiation of the electron beam is metered 25KGy.

[0021] 优选的,在所述表面活性剂溶液浸泡的步骤中,所述表面活性剂为TritonX_100、 Tween80或Tween40中的至少一种。 [0021] Preferably, in the step of soaking the surfactant solution, the surfactant is TritonX_100, Tween80 or Tween40 at least one of.

[0022] 优选的,所述动物膜组织原材料为出生6个月以下的猪或牛的膜组织材料。 [0022] Preferably, the animal tissue membrane material is less than 6 months of birth porcine or bovine tissue material film.

[0023] 所述磷酸盐缓冲液浸泡的步骤中,所述磷酸盐缓冲液的pH值为5.0-8.0。 The pH of step [0023] The immersion in phosphate buffer, said phosphate buffer is 5.0-8.0.

[0024] 优选的,在所述磷酸盐缓冲液浸泡的步骤中,所述浸泡后的膜组织加入pH为6.8的磷酸盐缓冲液中浸泡至少5h。 [0024] Preferably, in the step of soaking in a phosphate buffer, a film structure after the immersion was added to pH 6.8 phosphate buffer soaked at least 5h.

[0025] 本发明提供了一种由上述的方法制备得到的硬脑/脊膜生物补片材料。 [0025] The present invention provides dura one kind obtained by the above-described method for preparing / meninges biological patch material.

[0026] 本发明还提供了一种所述的硬脑/脊膜生物补片材料在制备防止或减少组织粘连材料、用于修复受损的硬脑/脊膜组织的材料、治疗或预防因外伤或肿瘤造成的硬脑/脊膜损伤或缺损的材料、治疗或预防因硬脑/脊膜缺损引起的脑脊液漏、感染材料中的应用。 [0026] The present invention further provides one of said dura / meningeal biological patch material in the preparation to prevent or reduce tissue adhesion material, a hard material for repairing damaged brain / meninges tissue, the treatment or prevention of brain trauma or hard meninges damage resulting from tumor or defective material, treating or preventing leakage of cerebrospinal fluid by hard brain / meninges caused by defects, infectious material application /.

[0027] 本发明还提供了一种缩水甘油醚类环氧交联剂用于制备硬脑/脊膜生物补片材料的用途。 [0027] The present invention also provides a glycidyl ether type epoxy crosslinking agent for the manufacture of hard brain / meninges of biological patch material.

[0028] 本发明所述的技术方案相比现有技术具有如下优势: [0028] The technical solution of the present invention has the following advantages over the prior art:

[0029] (1)本发明所述的硬脑/脊膜生物补片材料制备方法,利用质量浓度为l-5wt%缩水甘油醚类环氧交联剂浸泡处理膜组织材料,获得的生物补片材料特别适用于硬脑/脊膜缺损的修复,在植入人体或动物体内后,能够在体内保持6月-2年时间内稳定不降解,当超过2年后即降解,保证具有一定力学性能以及防液体渗漏作用的同时还具有生物相容性,产生术后并发症以及副作用的风险大大降低,满足了对于用于修复硬脑/脊膜缺损材料的特殊要求; [0029] (1) according to the present invention dura / meningeal patch preparation of the biological material, the use concentration of l-5wt% ether type epoxy crosslinking soaking membrane tissue, biological complement obtained sheet material is particularly suitable for the repair of hard brain / meninges defect, after implantation in the human or animal body, can be held in June in vivo - a stable non-degradable within 2 years, when more than 2 years after degradation, guarantee a certain mechanical and performance while also preventing liquid leakage effect biocompatible, risk of complications, and greatly reduces side effects, to meet the special requirements for repairing dura / mater defect material;

[0030] (2)本发明所述的硬脑/脊膜生物补片材料制备方法,通过选择丙三醇缩水甘油醚和/或乙二醇缩水甘油醚中为交联剂,不会增加该硬脑/脊膜生物补片本身的细胞毒性,降低其带来副作用的风险,在使用上述交联剂的补片,细胞毒性虽有略微提升,但是其提升幅度较小,可以忽略不计,并且其毒性都在〇级范围内,并不会对被植入的人体或动物产生不利的影响; [0030] (2) according to the present invention dura / meningeal biological patch material prepared by selecting a glycerol ether and / or ethylene glycol diglycidyl ether as a crosslinking agent, without increasing the hard brain / meninges biological cytotoxicity patch itself, which reduces the risk of side effects caused, in the patch using the crosslinking agent, a cytotoxic although slightly improved, but enhance its small amplitude, it can be ignored, and its toxicity are within the scope billion level, will not have an adverse effect on human or animal to be implanted;

[0031] (3)本发明所述的硬脑/脊膜生物补片材料制备方法,通过碱液以及表面活性剂配合使用获得的生物补片材料脂肪含量明显降低,大大提高了生物补片的安全性和有效性。 [0031] (3) according to the present invention dura / meningeal biological patch material prepared by the alkali solution and a surfactant complex biological patch material using the obtained fat decreased, greatly improving the bio-patch safety and efficacy.

附图说明 BRIEF DESCRIPTION

[0032] 为了使本发明的内容更容易被清楚的理解,下面根据本发明的具体实施例并结合附图,对本发明作进一步详细的说明,其中 [0032] In order to make the present invention understood more readily apparent, according to the following particular embodiments of the invention taken in conjunction with the accompanying drawings, the present invention will be further described in detail, wherein

[0033] 图1是未交联的膜材料的扫描电镜照片; [0033] FIG. 1 is a scanning electron micrograph of the film of uncrosslinked material;

[0034] 图2是交联的膜材料的扫描电镜照片; [0034] FIG. 2 is a scanning electron micrograph of the cross-linked film material;

[0035] 图3为植入未交联的生物补片的对照组照片; [0035] FIG. 3 is a set of photographs implanted control uncrosslinked biological patch;

[0036] 图4为植入实施例1制备的硬脑/脊膜生物补片的试验组的照片; [0036] FIG. 4 is a photograph implanted dura test group prepared in Example 1 / meninges biological patch;

[0037] 图5本发明实施例1制备的生物补片的HE染色图; FIG Biological HE staining [0037] 5 embodiment of the present invention prepared in Example 1 of the patch;

[0038] 图6为未使用硬脑/脊膜修复的对照组; [0038] FIG. 6 unused dura / meningeal repair of the control group;

[0039] 图7为使用实施例1制备的硬脑/脊膜生物补片修复的试验组。 [0039] Example 7 is prepared using an embodiment of the dura / meningeal biological test group patch repair.

具体实施方式 Detailed ways

[0040] 实施例1 [0040] Example 1

[0041] 本实施所述的硬脑/脊膜生物补片的制备方法包括如下步骤: [0041] The production method according to the present embodiment of the dura / meningeal biological patch comprising the steps of:

[0042] (1)取出生6个月以下的牛的膜组织为原材料,进行清洗,去除不适宜加工的部分, 备用; [0042] (1) 6 months of postnatal bovine membrane tissue as raw material, is cleaned to remove undesirable processing portion standby;

[0043] (2)取上述清洗后的膜组织加入质量浓度为0. lwt%的TritonXjOO溶液中浸泡, 震荡过夜,并控制溶液温度为〇°c; After the membrane tissue [0043] (2) to take the washing was added to 0. lwt% concentration solution of TritonXjOO soaking overnight shaking, and the solution temperature was controlled square ° c;

[0044] (3)取上述步骤浸泡后的膜组织加入浓度为5mol/L的氢氧化钠溶液中浸泡2h,并控制溶液温度为15 °C ; After the membrane tissue [0044] (3) take the above soaking step is added at a concentration of 5mol / L sodium hydroxide solution soaking 2h, and controlling the solution temperature was 15 ° C;

[0045] (4)取上述步骤浸泡后的膜组织加入质量浓度为lwt%过氧化氢溶液中浸泡处理lh; After the membrane tissue [0045] (4) take the above soaking step is added to lwt% concentration hydrogen peroxide solution, soaking LH;

[0046] (5)取上述步骤浸泡后的膜组织加入浓度为lwt%丙三醇缩水甘油醚中浸泡15h; After the membrane tissue [0046] (5) take the above soaking step is added at a concentration of lwt% glycerol polyglycidyl ether soaking 15H;

[0047] ⑶随后取所述浸泡后的膜组织加入pH为5.0的磷酸盐缓冲液中浸泡至少5h; [0047] ⑶ film is then taken after the tissue soaked added phosphate buffer pH 5.0 of at least 5H soaking;

[0048] (7)将上述步骤浸泡后的膜组织冷冻干燥,所述的冷冻干燥程序为:产品预冻先降温至-60°C以下后抽真空、升温,进入升华阶段;一次升华温度由_35°C以下升至_25°C,时间不少于6小时;一次升温温度由-25°C升至_15°C,时间为1-3小时;二次升温温度由-15°C升至25°C,时间为1-3小时。 [0048] (7) a film structure after the step of dipping the above-described lyophilized according to lyophilization procedure: pre-freezing the product after cooling to -60 ° C to less vacuum, heating, sublimation into the stage; sublimation temperature of _35 ° C or less was raised to _25 ° C, time less than 6 hours; once heated by the temperature rose to -25 ° C _15 ° C, the time is 1-3 hours; the secondary heating temperature -15 ° C raised to 25 ° C, the time is 1-3 hours. 随后以辐照计量为15KGy的电子束辐射灭菌,即得。 Followed by irradiation of electron beam radiation sterilization metering 15KGy, i.e., too.

[0049] 实施例2 [0049] Example 2

[0050] 本实施所述的硬脑/脊膜生物补片的制备方法包括如下步骤: [0050] The production method according to the present embodiment of the dura / meningeal biological patch comprising the steps of:

[0051] (1)取出生6个月以下的牛的膜组织为原材料,进行清洗,备用; [0051] (1) 6 months of postnatal bovine membrane tissue as raw materials, cleaning, standby;

[0052] (2)取上述清洗后的膜组织加入质量浓度为5wt%的TWeen80溶液中浸泡,震荡过夜,并控制溶液温度为25 °C ; After the membrane tissue [0052] (2) was added to take the cleaning mass concentration of 5wt% solution of TWeen80 soaking overnight shaking, and controlling the solution temperature 25 ° C;

[0053] (3)取上述步骤浸泡后的膜组织加入浓度为0. lmol/L的氢氧化钾溶液中浸泡4h, 并控制溶液温度为5 °C ; After the membrane tissue [0053] (3) take the above soaking step is added at a concentration of 0. lmol / L potassium hydroxide solution soak 4h, and controlling the solution temperature is 5 ° C;

[0054] (4)取上述步骤浸泡后的膜组织加入质量浓度为5wt%过氧化物溶液中浸泡处理5h; After the membrane tissue [0054] (4) take the above soaking step is added 5wt% of the peroxide concentration in the soaking solution, 5H;

[0055] (5)取上述步骤浸泡后的膜组织加入质量浓度为lwt%乙二醇缩水甘油醚中浸泡15h; [0055] (5) taken after the tissue membrane soaking step is added concentration of lwt% ethylene glycol diglycidyl ether soaking 15H;

[0056] ⑶随后取所述浸泡后的膜组织加入pH为8.0的磷酸盐缓冲液中浸泡至少5h; [0056] ⑶ film is then taken after the tissue soaked added phosphate buffer pH 8.0 of at least 5H soaking;

[0057] (7)将上述步骤浸泡后的膜组织冷冻干燥,所述的冷冻干燥程序为:产品预冻先降温至-60°C以下后抽真空、升温,进入升华阶段;一次升华温度由_35°C以下升至_25°C,时间不少于6小时;一次升温温度由-25°C升至_15°C,时间在1-3小时;二次升温温度由-15°C升至25°C,时间在1-3小时。 [0057] (7) a film structure after the step of dipping the above-described lyophilized according to lyophilization procedure: pre-freezing the product after cooling to -60 ° C to less vacuum, heating, sublimation into the stage; sublimation temperature of _35 ° C or less was raised to _25 ° C, time less than 6 hours; once heated by the temperature rose to -25 ° C _15 ° C, at the time of 1-3 hours; the secondary heating temperature -15 ° C raised to 25 ° C, at the time of 1-3 hours. 随后以辐照计量为25KGy的电子束辐射灭菌,即得。 Followed by irradiation of electron beam radiation sterilization metering 25KGy, i.e., too.

[0058] 实施例3 [0058] Example 3

[0059] 本实施所述的硬脑/脊膜生物补片的制备方法包括如下步骤: [0059] The production method according to the present embodiment of the dura / meningeal biological patch comprising the steps of:

[0060] (1)取出生6个月以下的牛的膜组织为原材料,进行清洗,备用; [0060] (1) 6 months of postnatal bovine membrane tissue as raw materials, cleaning, standby;

[0061] (2)取上述清洗后的膜组织加入质量浓度为2wt%的含有TritonX_100和Tween40 溶液中浸泡,震荡过夜,并控制溶液温度为8 °C ; After the membrane tissue [0061] (2) was added to take the cleaning mass concentration of 2wt% Tween40 solution containing TritonX_100 and soaking overnight shaking, and controlling the temperature of the solution is 8 ° C;

[0062] (3)取上述步骤浸泡后的膜组织加入浓度为lmol/L的氢氧化钠溶液中浸泡2h,并控制溶液温度为8°C ; [0062] (3) After taking the tissue membrane soaking step is added at a concentration of lmol / L sodium hydroxide solution soaking 2h, and controlling the temperature of the solution is 8 ° C;

[0063] (4)取上述步骤浸泡后的膜组织加入质量浓度为2wt%过氧化物溶液中浸泡处理3h; After the membrane tissue [0063] (4) take the above soaking step is added concentration of 2wt% peroxide solution soaking 3H;

[0064] (5)取上述步骤浸泡后的膜组织加入质量浓度为2wt%含有丙三醇缩水甘油醚和乙二醇缩水甘油醚的混合交联剂中浸泡20h; After the membrane tissue [0064] (5) take the above soaking step is added at a concentration of 2wt% mass containing the crosslinking agent ethylene glycol diglycidyl ether and glycerol polyglycidyl ether soaked for 20 h;

[0065] ⑶随后取所述浸泡后的膜组织加入pH为6.8的磷酸盐缓冲液中浸泡至少5h; [0065] ⑶ then taken after the immersion membrane tissue was added to pH 6.8 phosphate buffer at least soaked 5H;

[0066] (7)将上述步骤浸泡后的膜组织冷冻干燥,所述的冷冻干燥程序为:产品预冻先降温至-60°C以下后抽真空、升温,进入升华阶段;一次升华温度由_35°C以下升至_25°C,时间不少于6小时;一次升温温度由-25°C升至_15°C,时间在1-3小时;二次升温温度由-15°C升至25°C,时间在1-3小时。 [0066] (7) a film structure after the step of dipping the above-described lyophilized according to lyophilization procedure: pre-freezing the product after cooling to -60 ° C to less vacuum, heating, sublimation into the stage; sublimation temperature of _35 ° C or less was raised to _25 ° C, time less than 6 hours; once heated by the temperature rose to -25 ° C _15 ° C, at the time of 1-3 hours; the secondary heating temperature -15 ° C raised to 25 ° C, at the time of 1-3 hours. 随后以辐照计量为30KGy的电子束辐射灭菌,即得。 Followed by irradiation of electron beam radiation sterilization metering 30KGy, i.e., too.

[0067] 实施例4 [0067] Example 4

[0068] 本实施所述的硬脑/脊膜生物补片的制备方法包括如下步骤: [0068] The production method according to the present embodiment of the dura / meningeal biological patch comprising the steps of:

[0069] (1)取出生6个月以下的牛的膜组织为原材料,进行清洗,备用; [0069] (1) 6 months of postnatal bovine membrane tissue as raw materials, cleaning, standby;

[0070] (2)取上述清洗后的膜组织加入质量浓度为3wt%的含有TritonX_100和Tween80 溶液中浸泡,震荡过夜,并控制溶液温度为〇°c; After the membrane tissue [0070] (2) was added to take the cleaning mass concentration of 3wt% Tween80 solution containing TritonX_100 and soaking overnight shaking, and the solution temperature was controlled square ° c;

[0071] (3)取上述步骤浸泡后的膜组织加入浓度为lmol/L的氢氧化钠溶液中浸泡4h,并控制溶液温度为8°C ; [0071] (3) After taking the tissue membrane soaking step is added at a concentration of lmol / L sodium hydroxide solution soak 4h, and controlling the temperature of the solution is 8 ° C;

[0072] (4)取上述步骤浸泡后的膜组织加入质量浓度为4wt%过氧化物溶液中浸泡处理4h; After the membrane tissue [0072] (4) take the above soaking step is added concentration of 4wt% peroxide solution soaking 4H;

[0073] (5)取上述步骤浸泡后的膜组织加入质量浓度为5wt%含有丙三醇缩水甘油醚和乙二醇缩水甘油醚的混合交联剂中浸泡IOd; After the membrane tissue [0073] (5) take the above soaking step is added at a concentration of 5wt% mass containing the crosslinking agent ethylene glycol diglycidyl ether and glycerol polyglycidyl ether soaked IOd;

[0074] ⑶随后取所述浸泡后的膜组织加入pH为8.0的磷酸盐缓冲液中浸泡至少5h; [0074] ⑶ film is then taken after the tissue soaked added phosphate buffer pH 8.0 of at least 5H soaking;

[0075] (7)将上述步骤浸泡后的膜组织冷冻干燥,所述的冷冻干燥程序为:产品预冻先降温至-60°C以下后抽真空、升温,进入升华阶段;一次升华温度由_35°C以下升至_25°C,时间不少于6小时;一次升温温度由-25°C升至_15°C,时间在1-3小时;二次升温温度由-15°C升至25°C,时间在1-3小时。 [0075] (7) a film structure after the step of dipping the above-described lyophilized according to lyophilization procedure: pre-freezing the product after cooling to -60 ° C to less vacuum, heating, sublimation into the stage; sublimation temperature of _35 ° C or less was raised to _25 ° C, time less than 6 hours; once heated by the temperature rose to -25 ° C _15 ° C, at the time of 1-3 hours; the secondary heating temperature -15 ° C raised to 25 ° C, at the time of 1-3 hours. 随后以辐照计量为IOKGy的电子束辐射灭菌,即得。 Subsequently irradiating the measurement of electron beam radiation sterilization IOKGy, ie.

[0076] 对照例1 [0076] Comparative Example 1

[0077] 本实施例与实施例1的制备方法相同,其区别仅在于没有经过交联剂处理的步骤。 [0077] The present embodiment is the same as in Example 1 preparation method, except that only the step of cross-linking agent has not elapsed.

[0078] 对照例2 [0078] Comparative Example 2

[0079] 本实施例与实施例2的制备方法相同,其区别仅在于没有经过交联剂处理的步骤。 [0079] Example embodiments of the present embodiment is the same as the production method 2, except that only the step of cross-linking agent has not elapsed.

[0080] 对照例3 [0080] Comparative Example 3

[0081] 本实施例与实施例1的制备方法相同,其区别仅在于没有经过所述表面活性剂浸泡的步骤。 [0081] The embodiment of the present embodiment is prepared in the same manner as in Example 1, except that only the step of soaking said surfactant is not passed.

[0082] 对照例4 [0082] Comparative Example 4

[0083] 本实施例与实施例1的制备方法相同,其区别仅在于所用交联剂为戊二醛交联。 [0083] Example embodiment of the present embodiment is prepared in the same manner as in Example 1, except that only the glutaraldehyde crosslinked with a crosslinking agent.

[0084] 效果例 [0084] Effect Example

[0085] 以下通过实验例来考察本发明下述相关实施例及对照例制备的硬脑/脊膜生物补片材料各项性能,进而证明本发明制备的硬脑/脊膜生物补片具有显著技术效果。 [0085] The following embodiments related to examine the following experimental examples and the present invention is to dura biological properties of the Comparative Example prepared patch material / meninges, and further demonstrate the preparation of the present invention dura / meningeal patch has significant biological technical effect.

[0086] 1、撕裂力测试 [0086] 1, Tear Tests

[0087] 试验方法:使用拉伸(压缩)试验机,将按照实施例1-2、对照例1-2方法制备的生物补片材料,将样品剪成规格为lcmX3cm的长条状,在离短边边缘3-5mm处用4-0号缝合线穿过样品,对折缝合线,在距离穿孔处约5cm处将缝合线打结,防止缝合线脱落;将上述方法得到的试样,用纯化水水化3-5分钟,将未穿线的一端固定在拉力试验机的下部,将穿线一端通过挂钩固定于拉力试验机的上部,开始检测,直至试样被撕裂,读取拉伸负荷力的最大值,即为本试样的撕裂力。 [0087] Test Method: Tensile (compression) testing machine, according to Example 1-2, the biological patch material of Comparative Example 1-2 The method of preparation, the samples were cut into elongated lcmX3cm specifications, from the 3-5mm shorter side edge of the sample passes through a No. 4-0 suture, the suture is folded, at about 5cm from the suture knot perforation, preventing the suture off; the samples obtained as described above, with purified of water of 3-5 minutes, one end is fixed to the lower non-threaded tensile testing machine, the threading hook through an upper end fixed to the tensile tester starts to detect, until the specimen is torn, the tensile load force reading the maximum tearing force is the present sample. 结果见下表1以及图1-2。 Results Table 1 below and Figure 1-2.

[0088] 表1撕裂力测试结果 [0088] Tear Test Results TABLE 1

Figure CN104130437BD00081

[0090] 结论:上表为三次试验共30片产品撕裂力的平均值,实施例1、2无论15KGy还是25KGy的剂量下,相对于无交联剂处理的对照例1、2的情况,撕裂力均有明显提升,提升比例分别为54.79 %和44.90%,而相对于采用其他交联剂处理的对照例4,相对于无交联剂处理的对照例1、2的情况,同样撕裂力均有明显提升,提升比例分别为16.94%和36.19%。 [0090] Conclusion: The table above is a total of three tests tearing of the product 30 forces the average, at a dose of 25KGy to 15KGy whether or Examples 1 and 2, no cross-linking agent with respect to the processing of Comparative Example 1 and 2, tearing force has improved significantly enhance the proportion of 54.79% and 44.90%, respectively, relative to the use of other crosslinking agents for the treatment of Comparative Example 4, with respect to the case where no cross-linking agent of Comparative Example 1, the same tear cracking force has improved significantly enhance the proportion of 16.94% and 36.19%, respectively. 由于膜组织材料的胶原纤维无规编织而形成的纤维之间存在大量不规则的间隙,孔隙率主要取决天然胶原纤维的结构,交联剂会在胶原间形成新的键将胶原纤维拉紧引起膜材料的轻微收缩,从图1-2中对比可知,交联前后,生物补片的胶原纤维有明显的收缩,证明使用本发明的交联剂有交联效果,对膜的胶原纤维有一定的拉紧作用。 A large number of the fibers due to the irregular organization of collagen fibers intertwined at random material film formed by a gap, mainly depends on the porosity of the collagen structure of the natural fibers, the crosslinking agent will form a new bond between the collagen of the collagen fibers due to tension minor shrink film material, seen from comparison of FIGS. 1-2, before and after crosslinking, the collagen fibers are bio-patch shrink significantly, that the use of the present invention, the crosslinking agent has a crosslinking effect, collagen fiber membrane must the tightening effect.

[0091] 2、体外降解方面 [0091] 2. In vitro degradation aspects

[0092] 试验方法:将按照实施例1-2、对照例3-4方法制备的生物补片分剪成(1-3) * (4-6) mm大小的长条状,分别放置于试管中,加入一定量的生理盐水,放置在37°C中Id后,更换一次溶液,然后放置3-4d,取溶液根据医疗器械行业标准16886内的检测方法进行检测,试验结果见下表2。 [0092] Test Method: according to Example 1-2, 3-4 of a method of biological patch preparation of Comparative Example partial cut (1-3) * (4-6) mm elongated size, were placed in a test tube after the added amount of physiological saline, placed in a 37 ° C in Id, replacing the solution once, and then left 3-4d, take the solution is detected based on detection methods within the medical device industry standard 16886, the test results in Table 2 below.

[0093] 表2降解质量百分比 [0093] TABLE 2 mass percent degradation

Figure CN104130437BD00082

[0095] 上面表格表明分别为15KGy和25KGy辐照剂量生物补片的降解质量百分比,通过比较可以看出在有交联剂作用的情况下,生物补片的降解速度明显下降。 [0095] The above table shows that the degradation percentages are by mass 15KGy and 25KGy biological patch irradiation dose, it can be seen by comparing the effect of the crosslinking agent, the rate of biological degradation of the patch decreased.

[0096] 3、体内降解方面 [0096] 3, aspects of the in vivo degradation

[0097] 试验方法:将按照实施例1、对照例1、对照例4方法制备的生物补片,将上述制备的生物补片原位植入健康的大鼠体内,所述植入试验方法如下:取20只雄性SD大鼠,3-4周龄, 体重180-220g,麻醉后皮下植入实施例1的生物补片,在对侧植入对照例1或对照例4的生物补片,每张材料lcm2,选取植入3个周、12个周、30个周后各5只大鼠处死,取所述植入区的周围组织用多聚甲醛固定照相。 [0097] Test Method: The rats according to Example 1, Comparative Example 1, the patch biological method of Comparative Example 4 prepared in situ biological patch prepared above health implant, the implant following test methods : twenty male SD rats, 3-4 weeks old, weighing 180-220 g, were anesthetized and implanted subcutaneously biological Example 1 patch, on the opposite side of the implant or biological Comparative Example 1 Comparative Example 4 patch, each LCM2 material, selected implant three weeks, 12 weeks, 5 rats were sacrificed after 30 weeks, with paraformaldehyde fixed camera taking tissue surrounding the implant area. 试验结果见表3以及图3-4,图3为对照例1的植入12个周后的小鼠照片,图4为实施例1的植入12个周后的小鼠照片,从图中可以看出,图3已完全降解,图4还有未降解的生物补片存在,且膜形态比较完整,膜与周围组织未发生粘连情况,降解时间明显比未交联或选用其他交联剂制备的生物补片延长。 The results in Table 3 and FIGS. 3-4, FIG. 3 is a photograph of mice after implantation of 12 weeks of Comparative Example 1, FIG. 4 is a photograph of mice after implantation of 12 weeks Example 1 embodiment, from FIG. It can be seen in FIG. 3 has been completely degraded, the presence of FIG. 4 are not biodegradable as well as patches, film morphology and relatively complete, the film adhesions with the surrounding tissue does not occur, degradation time significantly higher than non-crosslinked or use other crosslinking agents biological extended patch preparation. 试验结果见下表3。 The results in Table 3 below.

[0098] 表3体内降解试验结果 [0098] Table 3 Test results of in vivo degradation

Figure CN104130437BD00091

[0100] 4、细胞毒性方面 [0100] 4, cytotoxicity

[0101] 试验方法:将按照实施例1、对照例1、对照例4的方法制备的生物补片进行浸提,制备浸提液;制备相应的细胞悬液;弃去原培养基,在96孔板中加入适当浓度的浸提液、阴性对照、阳性对照以及空白对照,置于37°C二氧化碳培养箱内分别培养24h、48h。 [0101] Test method: according to Example 1, Comparative Example 1, the biological method for the patch of Comparative Example 4 prepared leached, prepared extracts; prepared from the corresponding cell suspension; original medium was discarded, 96 well plate was added an appropriate concentration of the extract, negative control, positive control and blank control, placed in culture 24h, 48h, respectively, within 37 ° C carbon dioxide incubator. 在24h和48h 后,观察细胞生长状况,弃去96孔培养板内的浸提液,并加入lmg/ml的MTT溶液50yL。 After 24h and 48h, cell growth was observed conditions, discard the extract to the 96-well plate, and MTT solution was added 50yL lmg / ml of. 加入MTT溶液的96孔板置于37 °C二氧化碳培养箱2小时。 MTT solution was added to the 96 well plate was placed 37 ° C carbon dioxide incubator for 2 hours. 2小时后,弃掉96孔板中的MTT溶液,并加入异丙醇IOOyU震荡96孔板5-10min,测定其在492nm处的吸光度值。 After 2 hours, MTT solution was discarded 96 well plates, and isopropyl alcohol was added 96 volatility IOOyU 5-10min, absorbance values ​​measured at 492nm. 细胞毒性的检测方法是参照16886行业标准中细胞毒性检测中的MTT法进行的检测。 Cytotoxicity Detection detection reference is the industry standard 16886 Detection of cytotoxic MTT assay. 结果见下表4, The results in Table 4,

[0102] 表4细胞毒性试验结果 [0102] The results in Table 4 Cytotoxicity

Figure CN104130437BD00092

[0104] 上表为三次试验产品细胞毒性的平均值,可以清楚的看到实施例1使用交联剂制备的产品,细胞毒性有所降低,且都在0级范围内,相比于对照例1的细胞毒性实施例1的下降比例为2.80%和2.79%,细胞毒性变化不大,说明加入此交联剂后,不会增加产品本身的细胞毒性,然而相比于对照例1的细胞毒性对照例4的细胞毒性有一定的增加,且增加比例为9.01 %和11.46%,说明对照例4中所用交联剂有一定的细胞毒性。 [0104] The table above is three times the average cytotoxicity test product, can clearly see the product prepared in Example 1 embodiment using a crosslinking agent, decreased cytotoxicity, and are in the range of 0, compared to Comparative Example cytotoxicity decreasing rate of Example 1 of embodiment 1 is 2.80% and 2.79%, little change in cytotoxicity, indicating that the addition of this cross-linking agent, does not increase the cytotoxicity of the product itself, however, as compared to Comparative Example 1 cytotoxicity a certain increase of the cytotoxicity of Comparative Example 4, and increasing the proportion of 11.46% and 9.01%, in Comparative Example 4 described crosslinking agent has a certain cytotoxicity.

[0105] 5、脂肪含量方面 [0105] 5, the fat content aspect

[0106] 试验方法:将按照实施例1、对照例3方法制备的生物补片进行试验,脂肪含量的测定方法是依据《中华人民共和国药典》二部“胰酶脂肪检测”制定,测定结果见下表5。 [0106] Test Method: The test according to Example 1, a patch of biologically Comparative Example 3 prepared by the method, measurement method is based on the fat content of "Chinese Pharmacopoeia" two "fat pancreatin test" formulation, measurement results are table 5 below.

[0107] 表5脂肪含量试验结果 [0107] The results in Table 5 Fat content

Figure CN104130437BD00093

[0109] 上表为三次试验0. Ig生物补片脂肪含量的平均值,可以清楚的看到本发明实施例1通过碱液NaOH和表面活性性剂TrironX-IOO配合使用获得的生物补片,脂肪含量明显降低。 Is the average of three experiments in Table fat content of a biological patch 0. Ig [0109] on, can clearly see Example 1 of the present invention by lye NaOH and a surface active agent TrironX-IOO obtained with the use of bio-patch fat decreased significantly.

[0110] 6、安全性和有效性 [0110] 6. The safety and efficacy

[0111] 6.1安全性 [0111] 6.1 Security

[0112] 试验方法:将本发明实施例1制备的生物补片进行HE染色处理,所述HE染色处理步骤如下:将材料用梯度酒精下行入水,后用自来水和蒸馏水先后清洗,用0.5 %苏木素处理后用自来水清洗,再用1 %盐酸酒精分色后用自来水清洗,再用1/400的氨水促蓝后清洗,用1%伊红染色后用80%酒精分色,梯度酒精脱水,二甲苯透明后得到树脂封片。 [0112] Test Method: Biological patch prepared in Example 1 the embodiment of the present invention HE staining process, the HE staining following processing steps: the material into the water using a gradient alcohol down, after successively washing with tap water and distilled water, with 0.5% hematoxylin after treatment with tap water, then with 1% hydrochloric acid and alcohol after separation was washed with water, then aqueous ammonia after washing 1/400 pro blue, after staining with 1% eosin in 80% ethanol separation, gradient alcohol dehydration, two toluene to give a transparent resin mounted. 本发明实施例1制备的生物补片材料的HE染色图见图5,从图中可知,该生物补片没有细胞和细胞核残留,证明本发明的方法可以有效的脱去膜材料中的细胞等杂物,安全性高。 HE staining FIG biological patch material prepared in Example 1 of the present invention shown in Figure 5, seen from the drawing, the bio-patch no residual cells and nuclei, the method of the present invention may prove effective in the removal of the film material cells debris and safe.

[0113] 6.2有效性 [0113] 6.2 Effectiveness

[0114] 试验方法:将本发明实施例1制备的生物补片植入健康白兔中作为试验组,以未使用硬脑脊膜修复的兔子作为对照组,所述试验方法如下:麻醉兔子后,打开兔子头盖骨,破坏硬脑膜,加入生物补片替代硬脑膜,对照组不植入生物补片;然后将头盖骨复位,缝合伤口。 [0114] Test method: Biological patch prepared in Example 1 of the present invention implanted in healthy rabbits as the test group, is not used to repair dura rabbit as a control group, the test method is as follows: After the rabbit was anesthetized , rabbit skull is opened, damage the dura, the dura was added biological patch Alternatively, biological control group implanted patch; then reset the skull, the wound was sutured. 定期查看实验结果,试验结果见图6-7,图6为未使用硬脑脊膜修复的对照组,图7为使用本发明实施例1制备的硬脑/脊膜生物补片的试验组照片,从图中可以看出两张图片的伤口都已经痊愈,但图6照片中疤痕较大,且有部分组织出现坏死现象,而图7照片中伤口恢复良好,且疤痕较小,伤口平滑,没有组织坏死现象,说明修复组织功能良好且无免疫排斥反应。 Periodically check results, the test results shown in Figure 6-7, FIG. 6 is a control group not used to repair dura mater, FIG. 7 is used in Example 1 was prepared in the dura / meningeal patch embodiment of the present invention Biological tests set of photos from the figure can be seen in two pictures of the wounds have healed, but the scar 6 photos larger, and some tissue necrosis phenomenon, while 7 photos good wound healing and less scarring, wound smooth, no tissue necrosis, indicating good repair tissue function and no immune rejection.

[0115] 显然,上述实施例仅仅是为清楚地说明所作的举例,而并非对实施方式的限定。 [0115] Clearly, the above-described embodiments are merely made to clearly illustrate example, and not limited to the embodiment. 对于所属领域的普通技术人员来说,在上述说明的基础上还可以做出其它不同形式的变化或变动。 Those of ordinary skill in the art, on the basis of the above described variations or changes may be made in various other forms. 这里无需也无法对所有的实施方式予以穷举。 It is unnecessary and can not be exhaustive of all embodiments. 而由此所引伸出的显而易见的变化或变动仍处于本发明创造的保护范围之中。 The obvious changes or variations therefrom corollary is still in the scope of the inventions.

Claims (9)

1. 一种硬脑/脊膜生物补片材料的制备方法,其特征在于,包括取动物膜组织为原材料,并顺次置于表面活性剂溶液、碱液、过氧化物溶液、质量浓度为l-5wt%的缩水甘油醚类环氧交联剂溶液以及磷酸盐缓冲液中进行浸泡的步骤,并将经上述各步骤浸泡后的所述膜组织材料冷冻干燥后经辐照灭菌,即得。 A hard brain / meninges method for preparing a biological patch material, wherein the film comprises animal tissue taken as raw material, and sequentially placed in a surfactant solution, alkali solution, peroxide solution, the concentration of after soaking step l-5wt% of a glycidyl ether type epoxy crosslinking agent solution and phosphate buffer, and the tissue material by freezing the above-mentioned steps the film after soaking dried irradiation sterilization, i.e. too.
2. 根据权利要求1所述的硬脑/脊膜生物补片材料的制备方法,其特征在于,所述缩水甘油醚类环氧交联剂溶液浸泡的步骤中,所述交联剂为丙三醇缩水甘油醚和/或乙二醇缩水甘油醚,所述浸泡时间为15h-1 Od。 The method for preparing / meninges of biological dura patch material according to claim 1, wherein said step of glycidyl ether type epoxy crosslinking agent solution soaking, the crosslinking agent is a propionyl polyglycidyl ether and / or ethylene glycol diglycidyl ether, the soaking time is 15h-1 Od.
3. 根据权利要求1或2所述的硬脑/脊膜生物补片材料的制备方法,其特征在于,所述表面活性剂溶液浸泡的步骤中,所述表面活性剂的质量浓度为〇. l_5wt%,并控制所述溶液温度为0-25 °C进行浸泡。 3. dura or the method of preparation 1 / meninges biological patch material according to claim, wherein said step of soaking of the surfactant solution, the mass concentration of the surfactant is square. l_5wt%, and to control the temperature of the soaking solution to 0-25 ° C.
4. 根据权利要求1或2所述的硬脑/脊膜生物补片材料的制备方法,其特征在于,所述碱液浸泡的步骤中,所述碱液的浓度为0. l-5mol/L,并控制所述溶液温度为5-15°C进行浸泡。 The method for preparing / meninges of biological dura patch material of claim 1 or claim 2, wherein said step of soaking of alkali, the concentration of lye is 0. l-5mol / L, and controls the temperature of the soaking solution to 5-15 ° C.
5. 根据权利要求1或2所述的硬脑/脊膜生物补片材料的制备方法,其特征在于,所述过氧化物溶液浸泡的步骤中,所述过氧化物溶液的质量浓度为l-5wt%。 The method for preparing / meninges of biological dura patch material of claim 1 or claim 2, wherein the peroxide solution soak step, the mass concentration of the peroxide solution is l -5wt%.
6. 根据权利要求1或2所述的硬脑/脊膜生物补片材料的制备方法,其特征在于,所述磷酸盐缓冲液浸泡的步骤中,所述磷酸盐缓冲液的pH值为5.0-8.0。 The method for preparing / meninges of biological dura patch material of claim 1 or claim 2, wherein the phosphate buffer solution immersion step of, pH value of the phosphate buffer 5.0 -8.0.
7. 根据权利要求1或2所述的硬脑/脊膜生物补片材料的制备方法,其特征在于,所述辐照灭菌的步骤为电子束辐照灭菌,辐照计量为10_30KGy。 The method for preparing / meninges of biological dura patch material of claim 1 or claim 2, wherein the step of said radiation sterilization is electron beam irradiation sterilization, radiation measurement of 10_30KGy.
8. 根据权利要求1-7任一所述的方法制备得到的硬脑/脊膜生物补片材料。 Dura prepared according to any one of claims 1-7 obtained / meninges biological patch material.
9. 权利要求8所述的硬脑/脊膜生物补片材料用于制备防止或减少组织粘连材料、用于修复受损的硬脑/脊膜组织的材料、治疗或预防因外伤或肿瘤造成的硬脑/脊膜损伤或缺损的材料、治疗或预防因硬脑/脊膜缺损引起的脑脊液漏、感染材料中的用途。 9. The dura claimed in claim 8 / meninges cause biological patch material for the preparation to prevent or reduce tissue adhesion material, a hard material for repairing damaged brain / meninges tissue due to trauma or preventing or treating tumors hard brain / meninges damage or defective material, treating or preventing leakage of cerebrospinal fluid by hard brain / meninges caused by defects, the use of infected material.
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