CN100506295C - 生物衍生骨复合四环素缓释体 - Google Patents
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Abstract
本发明公开了一种生物衍生骨复合四环素缓释体及制备方法，该缓释体包括生物衍生骨支架、四环素和包被体，其特征是每克该缓释体含有62μg～122μg的四环素；生物衍生骨经四环素溶液浸泡，冷冻，干燥，包被，再冷冻干燥，消毒，即可制得生物衍生骨复合四环素缓释体，该缓释体具有比生物衍生骨具有更明显的成骨效应。体外释放盐酸四环素有效浓度可维持2周，临床应用安全，来源容易，价格低，易于推广使用。
Description
技术领域
本发明涉及骨组织修复的材料，特别是含有四环素的骨组织修复的材料。
背景技术
目前，用于促进骨组织修复材料的成骨能力的主要方式是复合促进骨生长的生长因子，其中研究最多的是骨形态发生蛋白系列(BMP2，4，7)和转化生长因子TGF-β，这些因子均为动物骨提取或经基因重组获得，除BMP2已被美国FDA批准用于临床外，其它均未被批准用于临床。因为促进骨生长的生长因子体内应用的最大问题是如何控制其有效剂量及担心过度表达带来的全身及局部问题，比如骨形态发生蛋白系列(BMP)由于使用剂量过大，过度表达，易导致植入部位过度增生，形成骨肉瘤，临床安全性受到质疑。
申请号为00132082.3，名称为“生物衍生组织工程骨及其制备方法”公开了三种生物衍生骨，为骨组织修复的材料，它们是将同种异体或异种(猪、牛)骨，经过物理化学、生物等方法处理后，去掉细胞、脂肪，部分蛋白或无机成份制得的一种生物材料，它是组织工程骨的支架，复合细胞后构建生物衍生组织工程骨，过去的研究已证实具有引导骨再生能力，临床用于骨组织修复，修复骨缺损。
四环素作为抗生素，临床应用已有近60年历史，1956年由Andro[1]发现了四环素的非抗生素作用。Lee(1988)[2]将四环素作为局部用药，Masato(1989)[3]将四环素缓释体置入牙周袋内治疗牙周病；20世纪中期发现了四环素对骨的作用，有研究者认为四环素可抑制骨生长，但更多的研究认为，仪在使用大剂量时才会出现骨生长抑制[4、5]。Sasaki(1974)[6]的研究发现四环素类在体内可防止骨密度的丢失、类骨质的丢失以及成骨细胞形态的改变。Saskki等[7]报告四环素(TCs)及其无抗生素作用的化学异构衍生物(CMTs)可完全恢复糖尿病鼠的成骨细胞形态和功能，并增加骨基质中H-脯氨酸标记的胶原合成。Aoyagi[8]等报告CMT治疗卵巢切除术后小鼠骨质疏松，可促进小梁骨和皮质骨内膜的新骨形成。
TCs、CMTs可提高成骨细胞活性，具有促进骨形成的作用[9]，这种作用是由于成骨细胞对四环素的明显摄取刺激成骨细胞分泌胶原所致。更为重要的是，四环素类药物在一系列的研究中证明其有明显的抑制骨吸收作用。体外实验发现，四环素可有效抑制由甲状旁腺激素(PTH)(Gomes et al 1984[10])、前列腺素E2(PGE E2)或脂多糖(LPS)(Golub et al 1984[11])刺激造成的骨吸收，并且降低成骨细胞来源的胶原酶活性[12、13]。体内实验中，四环素类也表明其抗骨质吸收的特性，Golub[14]、Grevstad[15]等报告四环素类可抑制细菌所致和手术所致的牙冠骨丢失，恢复糖尿病所致的全身骨质疏松。等等。
上述报道仅仅在药理机制上对四环素对骨的作用进行了探讨，并没有说明四环素成骨的量效关系，更没有将四环素实际应用于制备和生产骨组织修复材料产品中的报道。
发明内容
为了解决上述问题，本发明提供了一种骨组织修复的材料，它是生物衍生骨复合四环素缓释体，其包括生物衍生骨支架，四环素和包被体，以及生物衍生骨复合四环素缓释体的制备方法。
本发明解决其技术问题所采用的技术方案是：一种生物衍生骨复合四环素缓释体，其特征在于生物衍生骨中含有四环素。
所述的生物衍生骨，是专利申请号为00132082.3，名称为“生物衍生组织工程骨及其制备方法”所称的生物衍生骨支架，是用猪或异体骨采用物理化学方法制备三类衍生组织工程骨支架的任意一种，即：部分脱蛋白生物衍生骨支架、完全脱蛋白生物衍生骨支架和部分脱钙生物衍生骨支架。
所述的四环素为盐酸四环素，其中每克生物衍生骨复合盐酸四环素缓释体含有62μg～122μg的盐酸四环素，优选为80μg～110μg的盐酸四环素。
生物衍生骨复合四环素缓释体的制备方法，它包含下列步骤：先将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素溶液中；再将该生物衍生骨支架冷冻干燥，即得。
当所述的盐酸四环素溶液的溶剂为水时，该生物衍生骨复合四环素缓释体的制备方法，包括如下步骤：
a、将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素水溶液中，盐酸四环素的浓度为50μg/ml～100μg/ml，使每克生物衍生骨复合盐酸四环素缓释体含有62μg～122μg的盐酸四环素：
b、将a步骤的盐酸四环素水溶液在负压条件下浸泡；
c、将b步骤浸泡的生物衍生骨支架冷冻干燥；
d、包被：将c步骤的冷冻干燥物放入包被的物质中进行包被；
e、将d步骤已包被物质冷冻干燥，消毒，即得。
所述方法步骤d中用于包被的物质为藻酸钠，pluronicF-127(氧化异丙烯，无毒、无粒子表面活性，水溶液可形成凝胶，4℃以下由凝胶变为液态，高于15℃又转变为凝胶，体内可完全降解)、或聚乳酸。进一步地，d步骤用于包被的物质为藻酸钠，浓度为1％～2％w/v，pH7.2。
当所述盐酸四环素溶液的溶剂为pluronicF-127，浓度为25％w/v时，该生物衍生骨复合四环素缓释体的制备方法，包括如下步骤：
a、将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素pluronicF-127溶液中，盐酸四环素的浓度为50μg/ml～100μg/ml；
b、将a步骤的盐酸四环素溶液在负压条件下浸泡；
c、将b步骤浸泡的生物衍生骨支架冷冻干燥；
d、将c步骤已包被物质冷冻干燥，消毒，即得。
上述两种生物衍生骨复合四环素缓释体的制备方法中，步骤b所述的生物衍生骨支架在负压条件为：负压0.5mPa。
本发明的有益效果是：本发明通过将专利申请号为00132082.3，名称为“生物衍生组织工程骨及其制备方法”所述的生物衍生骨支架与四环素(盐酸四环素)制备成本发明生物衍生骨复合盐酸四环素缓释体，当盐酸四环素的用最为：每克生物衍生骨复合盐酸四环素缓释体含有62μg～122μg的盐酸四环素时，具有与促进骨生长的生长因子rhBMP、TGF-β基本相同的成骨效应，且克服了促进骨生长的生长因子的副作用，通过生物衍生骨支架与盐酸四环素组合使用，体外释放盐酸四环素有效浓度可维持2周，充分达到缓释功效，临床应用安全，来源容易，价格低，易于推广使用。
显然，根据本发明的上述内容，按照本领域的普通技术知识和惯用手段，在不脱离本发明上述基本技术思想前提下，还可以做出其它多种形式的修改、替换或变更。
以下通过实施例形式的具体实施方式，对本发明的上述内容再作进一步的详细说明。但不应将此理解为本发明上述主题的范围仪限于以下的实例。凡基于本发明上述内容所实现的技术均属于本发明的范围。
附图说明
图1：生物衍生骨复合四环素缓释体；
图2：复合四环素生物衍生骨复合成骨细胞培养7天。
具体实施方式
通过下面给出的本发明的实施例可以进一步清楚地了解本发明。但它们不是对本发明的限制。
实施例1本发明生物衍生骨复合四环素缓释体的制备
本发明生物衍生骨复合四环素缓释体是由生物衍生骨支架与四环素(盐酸四环素)制备而成。以下是详细的制备方法：
①生物衍生骨(即专利申请号为00132082.3的在先申请中所称的生物衍生骨支架)的制备
三类生物衍生骨支架的制备：
a、制备部分脱蛋白生物衍生骨支架的方法是：取新鲜猪骨或异体骨，去除其上所附软组织、中心骨髓及软骨部分，蒸馏水冲洗干净，于38℃30％过氧化氢中脱脂72小时，每24小时换液一次；蒸馏水浸洗干净，于75％酒精中去除残余过氧化氢24小时，蒸馏水浸洗干净；氯仿：甲醇3∶1，于室温下部分脱蛋白4小时，蒸馏水清洗；在—38℃，10-5Pa条件下冷冻干燥24小时；25KGYγ射线辐射处理消毒并清除剩余胶原蛋白的抗原性，最后获得部分脱蛋白生物衍生骨支架；
b、全脱蛋白生物衍生物骨支架的方法是：取新鲜猪骨或异体骨，去除其上所附软组织、中心骨髓及软骨部分，蒸馏水冲洗干净，于38℃30％过氧化氢中脱脂48小时，每24小时换液一次；蒸馏水浸洗干净，氯仿：甲醇3∶1，于室温下部分脱蛋白4小时，蒸馏水清洗；入110℃乙二胺中循环提取24小时，蒸馏水浸洗干净，于室温下入乙醇中浸泡24小时除去残余乙二胺，蒸馏水清洗；在—38℃，10-5Pa条件下冷冻干燥24小时；于20～50℃下经环氧乙烷灭菌3～9小时，最后获得完全脱蛋白生物衍生骨支架。
c、制备部分脱钙生物骨支架的方法是：取新鲜猪骨或异体骨，去除其上所附软组织、中心骨髓及软骨部分，蒸馏水冲洗干净，于38℃30％过氧化氢中脱脂48小时，每24小时换液一次；蒸馏水浸洗干净，入0.6M盐酸在4℃下部分脱钙72小时，每24小时换液一次；蒸馏水浸洗干净，氯仿∶甲醇3∶1，于室温下部分脱蛋白4小时，蒸馏水清洗；在—38℃，10-5Pa条件下冷冻干燥24小时；25KGYγ射线辐射处理消毒并清除剩余胶原蛋白的抗原性。最后获得脱钙生物衍生骨支架。
所制备的上述三类生物衍生骨支架的任意一种，保存了原来骨组织的基本框架，具有一定的生物力学功能，基本去除抗原性；
②生物衍生骨复合盐酸四环素缓释体的制备：
方法一：取盐酸四环素粉剂溶于蒸馏水中，使其浓度为0.05mg/ml。将已消毒灭菌的生物衍生骨材料放入溶液中，在4℃条件下放置48小时，取出后放入-80℃深低温冰箱中，冻存30分钟，或在4℃条件下保存48小时，再经冷冻干燥机处理，形成含盐酸四环素的复合物。取藻酸钠干粉配制成1％～2％溶液，pH7.2，高压灭菌备用。将已制备好的含盐酸四环素的复合物置于藻酸钠溶液中浸泡24～48小时，取出后经-80℃深低温冰箱保存30分钟，然后再经冷冻干燥处理，消毒备用。
方法二：配制25％pluronic F-127溶液，在其中加入盐酸四环素，在避光条件下溶解，使溶液中含盐酸四环素的终浓度为0.05mg/ml。将已制备好的生物衍生骨置于上述溶液中浸泡1小时后取出，经蒸馏水洗涤3次，-80℃深低温冰箱冷冻30分钟，然后经冷冻干燥处理后，包装、辐照(或环氧乙烷)消毒处理、保存。材料孔径平均为522±16.7um，孔隙率为55％，含水量为5％。
体外释放盐酸四环素有效浓度可维持2周，第3周完全消失。
实施例2比较四环素的给药方式(口服、静脉输入和生物衍生骨复合盐酸四环素缓释体)对成骨的影响
将静脉注射用四环素粉剂(上海新亚制药厂生产)作为一种促进骨生长的生长因子作为对照组。
在剂量为每克本发明实施例1制备的生物衍生骨复合盐酸四环素缓释体含有80μg～110μg的盐酸四环素时，碱性磷酸酶(ALP)活力为1.758±0.127IU/ml，对照组为1.251±0.144IU/ml，ALP活性增强，I型胶原合成能力增强，I型胶原mRNA表达、骨钙素及骨钙素mRNA表达增加，对新骨形成进行评分[16]：8周时为3.67，对照组为1.33，统计学处理具有显著性差异(P<0.05)。评分标准见表1。
表1 Lan-Sandhn X线评分标准
| 临床表现 | 分值 | |
| 骨形成 | 无 | 4 |
| 形成25％ | 1 | |
| 形成50％ | 2 | |
| 形成75％ | 3 | |
| 完全形成 | 4 | |
| 骨连接 | 骨折线清晰 | 0 |
| 部分存在 | 2 | |
| 折线消失 | 4 | |
| 骨塑形 | 无 | 0 |
| 髓腔形成 | 2 | |
| 皮质骨塑形 | 4 |
实施例3 生物衍生骨复合盐酸四环素缓释体与生物衍生骨复合rhBMP/TGF-β生长因子缓释体成骨效果比较
(1)在36只新西兰兔双后肢肌肉内植入的体内研究证实，将本发明实施例1制备的生物衍生骨复合盐酸四环素缓释体，与含rhBMP/TGF-β生长因子的复合物制成的缓释体对照，促进成骨的结果基本一致，从组织学看，植入后第8周，两组均形成混合性新骨，12周时已有编织骨形成，支架材料已完全降解吸收。生物衍生骨复合盐酸四环素缓释体略优于rhBMP/TGF-β组的成骨效果。单纯支架材料在整个实验过程中均未见到新骨形成。
(2)在此基础上进一步接种相同数量的成骨细胞，发现均有良好的细胞相容性。在体外实验中发现，这种缓释体有较单纯材料更多的细胞粘附率。经体外培养后植入体内，观察其成骨能力，骨钙素及I型胶原均有显著增加。血清骨钙素测定结果见表2
表2 血清骨钙素测定结果(ng/ml)
| 术前 | 2周 | 4周 | 8周 | 12周 | |
| 生物衍生骨 | 2.43±0.64 | 2.56±0.44 | 3.22±0.52 | 3.63±0.48 | 4.08±0.55 |
| 生物衍生骨+细胞 | 2.35±0.48 | 5.91±1.05 | 9.16±1.51 | 8.00±1.77 | 7.04±1.58 |
| 生物衍生骨+四环素 | 2.58±0.49 | 2.36±0.56 | 3.36±0.53 | 5.24±0.83 | 5.33±1.16 |
| 生物衍生骨+四环素+细胞 | 2.48±0.58 | 8.36±1.45 | 11.62±3.2 | 10.69±2.71 | 7.37±2.47 |
| H空白对照 | 2.53±0.61 | 2.31±0.39 | 2.88±0.52 | 3.01±0.83 | 2.55±0.76 |
植入本发明实施例1制备的生物衍生骨复合盐酸四环素缓释体动物在术后8、12周骨钙素含量显著高于生物衍生骨组(p<0.05)，植入复合成骨细胞的四环素生物衍生骨缓释体动物在术后2、4、8周骨钙素含量显著高于复合成骨细胞的生物衍生骨组(p<0.05)。表明复合盐酸四环素的生物衍生骨比未复合盐酸四环素的生物衍生骨具有更好的修复作用。
实施例4 本发明生物衍生骨复合盐酸四环素缓释体骨缺损修复实验
将pluronicF-127与盐酸四环素复合后制成缓释体，修复兔桡骨2cm缺损，发现其骨修复能力显著优于单纯生物衍生骨支架。在36只新西兰兔的实验研究中证实，生物衍生骨复合盐酸四环素缓释体组较单纯材料成骨的组织学表现提前7～14天，骨修复的生物力学强度表现出显著性差异。
1、三点弯曲模量测定(表3)
表3 三点弯曲模量(Mpa)
| 2周 | 4周 | 8周 | 12周 | |
| 生物衍生骨 | 18.14±7.21 | 55.42±8.37 | 122.76±11.51 | 142.03±12.71 |
| 生物衍生骨+细胞 | 42.53±6.67 | 134.52±14.74 | 166.37±17.22 | 157.48±16.65 |
| 生物衍生骨+四环素 | 20.1±9.33 | 64.63±13.24 | 144.79±18.82 | 148.54±19.15 |
| 生物衍生骨+四环素+细胞 | 57.86±10.29 | 135.53±11.18 | 182.55±16.25 | 158.44±16.12 |
植入四环素生物衍生骨缓释体动物在术后2、4、8周三点弯曲模量显著高于生物衍生骨组(p<0.05)，植入复合成骨细胞的四环素生物衍生骨缓释体动物在术后2、8周三点弯曲模量显著高于复合成骨细胞的生物衍生骨组(p<0.05)。
2、抗压强度、压缩横量测定(表4)
表4 抗压强度、压缩横量
| 2 | 4 | 8 | 12 | |
| 生物衍生骨 | 0.22±0.05 | 0.33±0.08 | 7.67±0.95 | 38.65±3.77 |
| 生物衍生骨+细胞 | 1.37±0.21 | 12.24±1.71 | 43.46±3.26 | 44.06±2.61 |
| 生物衍生骨+四环素 | 0.24±0.06 | 0.55±0.36 | 12.71±1.66 | 50.35±4.72 |
| 生物衍生骨+四环素+细胞 | 1.44±0.46 | 14.34±1.92 | 52.13±5.31 | 46.48±4.43 |
植入四环素生物衍生骨缓释体动物在术后2、4、8、12周压缩模量显著高于生物衍生骨组(p<0.05)，植入复合成骨细胞的四环素生物衍生骨缓释体动物在术后4、8周压缩模量显著高于复合成骨细胞的生物衍生骨组(p<0.05)。
上述实施例动物实验说明复合盐酸四环素的生物衍生骨比未复合盐酸四环素的生物衍生骨具有更好的作用。
参考文献：
1、Andro T Studies of the distribution of tritium labeled dihydrostrostreptomycin and tetracycline in thebody.Acta Radiol，(suppl)1956；142：1
2、Lee silverstein，Nabil Bissada M，Manouchehr-pour.Clinical periodontitis.J Peridontal，1988；59：301-305
3、Masato Minabe，Atsuo uematsu，Kayo Nishijima，et al.Application of a localrrugdelivery system toperiodontal therapy.J Periodontol，1989；60：113-117
4、Proffit WR，Bennett IC.Effects of tetracycline on bone growth in growth in organ culture.Growth，1968；32：113
5、Yen PDJ，Shaw JH.Preliminary study of inhibitory effects of tetracyclines on membranous bonegrowth in rhesus monkeys.J Dent Res，1978；51：1651
6、Sasaki T，Ramamurthy NS，Golub LM.Bone cell and matrix bind chemically modifiednonantimicrobial tetracycline.Bone，1994；15：373-375
7、Sasaki T，Kaneko H，Nungavaramt S，et al.Tetracycline administration restores osteoblast structure andfunction during experimental diabetes.The Anatomal Recore，1991；231：25-24
8、Aoyagi M，Sasdki T，Ramamurthy NS，et al.Tetracycline/Flurbiprofen combination therapy modulatesbone remodeling in ovariectomized rat：preliminary observations.Bone，1996；19：629-635
9、Golub LM<Ramamurthy NS，Kaneko H，et al.Tetracycline administration prevents diabetes-inducedosteopania in the rat：initial observations.Res Commun Chem Pathol Pharmacol，1990；68：24-40
10、Gomes BC，Golub LM，Ramamuthy NS.Tetracyclines inhibit parathyroid-induced bone resorption inorgan culture.Experientia，1984；40：1273-1275
11、Golub LM，Ramamurthy NS<McNamara TF，et al.Tetracyclines inhibit tissue collagenase activity：anew mechanism in the treatment of periodontal disease.Journal of Periodontal Research，1984；19：651-655
12、Ramamurthy NS，Vernillo AT，Lee HM，et al.Theeffect of tetracy clines on collagenase activity inMUR 106-01 rat osteoblastic osteosarcoma cells.Res Comnun Chem Pathol Pharmacol.1990：70：323-335
13、Ramamurthy NS.Vernillo AT，Grcenwald RA.Reactive oxygen specics activate and tetracyclinesinhibit rat osteoblast collagenase.J Bone Miner Res，1993；8：1247-1253
14、Golub LM，Lee HM，Lehrer G，et al，Minocycline reduces gingival collagenolytic activity duringdiabetes：preliminary observations and a proposed new mechanism of action.Journal of PeriodobnrtalResearch 1983，3(8)：516-524
15、G revestad，et al.Doxycycline prevents root resorption and alveolar bones loss in rat afterperiodontal surgery.Scand Dent Res，1993；99：411-414
16、Lan-Sandhu Lane，JM.，Tomin E，Mathiac PG.Biosynthetic boneg raft.Clin Orthop RelateResearch.1999：107-117
Claims (18)
1.一种生物衍生骨复合四环素缓释体，其特征在于每克生物衍生骨复合四环素缓释体中含有62ug～122ug的盐酸四环素。
2、根据权利要求1所述生物衍生骨复合四环素缓释体，其特征在于：每克生物衍生骨复合四环素缓释体含有80ug～110ug的盐酸四环素。
3、权利要求1所述的生物衍生骨复合四环素缓释体的制备方法，它包含下列步骤：先将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素溶液中；再将该生物衍生骨支架冷冻干燥，即得。
4、根据权利要求3所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：所述的盐酸四环素溶液的溶剂为水。
5、根据权利要求4所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：它包括如下步骤：
a、将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素水溶液中，盐酸四环素的浓度为50ug/ml～100ug/ml，使每克生物衍生骨复合四环素缓释体含有62ug～122ug的盐酸四环素，浸渍时的盐酸四环素水溶液处在负压条件下；
b、将a步骤浸泡的生物衍生骨支架冷冻干燥；
c、包被：将b步骤的冷冻干燥物放入包被的物质中进行包被；
d、将c步骤已包被物质冷冻干燥，消毒，即得。
6、根据权利要求5所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：所述方法步骤c中用于包被的物质为藻酸钠、pluronicF-127或聚乳酸。
7、根据权利要求6所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：c步骤用于包被的物质为藻酸钠，浓度为1％～2％w/v，pH7.2。
8、根据权利要求3所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：所述盐酸四环素溶液的溶剂为pluronicF-127，浓度为25％w/v。
9、根据权利要求8所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：它包括如下步骤：
a、将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素pluronicF-127溶液中，盐酸四环素的浓度为50ug/ml～100ug/ml，使每克生物衍生骨复合四环素缓释体含有62ug～122ug的盐酸四环素，浸渍时的盐酸四环素溶液处在负压条件下；
b、将a步骤浸泡的生物衍生骨支架冷冻干燥；
c、将b步骤已干燥物消毒，即得。
10、根据权利要求5或9所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：步骤b所述的生物衍生骨支架冷冻干燥在负压0.5mPa下进行。
11、权利要求2所述的生物衍生骨复合四环素缓释体的制备方法，它包含下列步骤：先将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素溶液中；再将该生物衍生骨支架冷冻干燥，即得。
12、根据权利要求11所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：所述的盐酸四环素溶液的溶剂为水。
13、根据权利要求12所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：它包括如下步骤：
a、将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素水溶液中，盐酸四环素的浓度为50ug/ml～100ug/ml，使每克生物衍生骨复合四环素缓释体含有的盐酸四环素为80ug～110ug，浸渍时的盐酸四环素水溶液处在负压条件下；
b、将a步骤浸泡的生物衍生骨支架冷冻干燥；
c、包被：将b步骤的冷冻干燥物放入包被的物质中进行包被；
d、将c步骤已包被物质冷冻干燥，消毒，即得。
14、根据权利要求13所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：所述方法步骤c中用于包被的物质为藻酸钠、pluronicF-127、或聚乳酸。
15、根据权利要求14所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：c步骤用于包被的物质为藻酸钠，浓度为1％～2％w/v，pH7.2。
16、根据权利要求11所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：所述盐酸四环素溶液的溶剂为pluronicF-127，浓度为25％w/v。
17、根据权利要求16所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：它包括如下步骤：
a、将已消毒灭菌的生物衍生骨支架在0～4℃浸渍于盐酸四环素pluromc F-127溶液中，盐酸四环索的浓度为50ug/ml～100ug/ml，使每克生物衍生骨复合四环素缓释体含有80ug～110ug的盐酸四环素，浸渍时的盐酸四环素溶液处在负压条件下；
b、将a步骤浸泡的生物衍生骨支架冷冻干燥；
c、将b步骤已干燥物消毒，即得。
18、根据权利要求13或17所述的生物衍生骨复合四环素缓释体的制备方法，其特征在于：步骤b所述的生物衍生骨支架冷冻干燥在负压0.5mPa下进行。
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