壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶的体外抗结核作用

文题释义：基质细胞衍生因子1：是一种参与免疫细胞活化、分化和迁移及伤口愈合、角膜上皮再生和组织修复等过程的趋化因子,能促进干细胞的生长和发育,参与调节成骨分化,可通过细胞归巢提高干细胞向病灶区的趋化作用。而基质细胞衍生因子1的失活会损害成骨细胞的发育和分化。此外,其还与血管生成密切相关。 异烟肼：具有较高的杀菌活性,是治疗结核病的一线药物。世卫组织建议将异烟肼作为结核病的标准疗法,用于潜伏性结核病感染者的预防治疗,与利福平、吡嗪酰胺和乙胺丁醇一起用于治疗活动性肺结核。异烟肼的活化形式与脂肪酸生物合成Ⅱ型系统中的NADH依赖型烯醇酰基载体蛋白还原酶异烟肼a结合,阻断细菌细胞壁关键成分支原体酸的合成。 背景：抗结核化疗是目前治疗骨关节结核的主要手段,然而全身给药难以维持病灶区的有效浓度,治疗效果欠佳。 目的：制备一种原位、长期释放抗结核药物且兼备促成骨作用的壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶。 方法：将亲水性的抗结核药物异烟肼和疏水性的基质细胞衍生因子通过复乳法负载到聚乳酸-羟基乙酸中,制备聚乳酸-羟基乙酸载药微球,共混至壳聚糖-明胶水凝胶支架中,制备壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶。检测聚乳酸-羟基乙酸载药微球、壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶的体外释药与抗结核杆菌的能力。将成骨前体细胞MC3T3-E1分别接种于载药微球与联合载药水凝胶表面,CCK-8法检测细胞活力,碱性磷酸酶活性检测细胞的成骨性能。 结果与结论：①载药微球中异烟肼1 h内的突释约为23.3%,2 d内的释放率约为42.6%,随后进入缓释期,25 d后进入平台期;基质细胞衍生因子1在1 h内的累积释放率约为19.8%,2 d内的释放率约为44.7%,随后进入缓释期,25 d后进入平台期;联合载药水凝胶中异烟肼和基质细胞衍生因子1最初1 h的释放分别为8.3%和8.5%,第2天的累计释放率分别为15.2%和17.6%,远低于聚乳酸-羟基乙酸微球;②体外4周后,联合载药水凝胶的抑菌直径大于载药微球,抑菌率高于载药微球(P < 0.05);③联合载药水凝胶与载药微球均具有良好的细胞相容性,细胞活力均约为100%;④培养5,10 d后,联合载药水凝胶表面的细胞碱性磷酸酶活性与载药微球比较差异无显著性意义(P > 0.05);⑤结果表明,原位壳聚糖-明胶/聚乳酸-羟基乙酸联合载药水凝胶有作为治疗骨关节结核及其他骨关节感染的潜力。 ORCID: 0000-0003-4166-2492(张贺龙) 中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程     

壳聚糖微球药物释放机制研究进展

近年来已有多种药物实现了以壳聚糖微球作为缓控释载体,并在生物医学领域展现出良好的应用前景,成为缓控释剂型研究的热点之一。目前对壳聚糖微球释放机制的研究进展落后于壳聚糖载药微球制备与应用的研究进展,而加强壳聚糖载药微球药物释放机制的研究,有利于更好地了解药物的释放行为和释放影响因素,并对深入研究壳聚糖缓释载药体系的制备与应用具有重要意义。主要从壳聚糖微球的药物释放机制、药物释放行为描述、药物释放影响因素等方面进行了综述。     

Recoverability of freeze-dried doxorubicin-releasing chitosan embolic microspheres

The purpose of this study is to investigate the recoverability of freeze-dried chitosan microspheres (MS). The factors influencing the integrity of chitosan MS during freeze-drying and rehydration procedures were determined, with focusing on choosing a suitable rehydration method and a freeze-drying excipient. Mean MS size, size distribution and sphericity of recovered chitosan MS were evaluated. Furthermore, the impacts of freeze-drying and rehydration procedure on the elasticity of chitosan MS were explored and the release profiles were evaluated. The recoverability of lyophilized chitosan MS was largely dependent on rehydration method and freeze-drying excipients. When using the optimized recovery processes, deformable drug-loaded chitosan MS can be rapidly recovered to exhibit the initial physico-mechanical properties such as elasticity. Release profiles also were not significantly changed after rehydration procedure. It is therefore expected drug-loaded chitosan MS can be stably freeze-dried with the prevention of drug release during storage and rapidly recovered to be used as deformable embolic materials possibly applicable for anti-cancer embolotherapy.     

Manufacturing monodisperse chitosan microparticles containing ampicillin using a microchannel chip

The purpose of this study was using a developed microfluidic chip to prepare size-controlled monodisperse chitosan microparticles encapsulating ampicillin. Our strategy is that a chitosan aqueous solution (the disperse phase) is fed into the microfluidic chip equipped with a cross-junction microchannel, and is sheared by the viscous oil flows (the continuous phase) to form monodisperse semi-product, chitosan emulsions. These fine emulsions are then gelled into stability upon gelation by injection of copper sulfate solution at the terminal microchannel of the microfluidic chip, and finally the uniform chitosan microparticles are formed in an efficient manner. The proposed chip is fabricated by a CO₂ laser machine on a conventional poly methyl methacrylate (PMMA) substrate. This microfluidic chip has four inlet ports, one cross-channel and one outlet port. We have demonstrated that one can control the size of chitosan microparticles from 100 to 800 μm in diameter (with a variation less than 5%) by altering the relative sheath/sample flow rate ratio. Experimental data showed that when given a steady continuous phase (oil flow), the emulsion size increases with the increase in average velocity of the dispersed phase flow (sample flow). In addition, the release of the model drug (ampicillin) from these microspheres is proved to be once-daily for clinical application. We also revealed that appropriate particle sizes for different release patterns are predictable, enabling better applications of chitosan as a drug carrier.     

戊二醛交联对壳聚糖/羟基磷灰石-庆大霉素缓释材料性能的影响

背景：交联是骨组织工程材料改性的一种常用方法,但目前仍缺乏交联剂对载药人工骨材料性能影响的相关研究与报道。 目的：研究戊二醛交联对壳聚糖/羟基磷灰石-庆大霉素载药人工骨材料力学性能、降解性能及体外药物缓释行为的影响。 方法：分别制备壳聚糖质量分数为10%,20%,30%的壳聚糖/羟基磷灰石-庆大霉素载药人工骨材料与戊二醛交联壳聚糖/羟基磷灰石-庆大霉素载药人工骨材料,检测各组材料的机械强度、吸水率、降解率及体外药物释放行为。 结果与结论：壳聚糖含量为10%,20%,30%壳聚糖/羟基磷灰石-庆大霉素的抗压强度分别为(10.16±1.17),(28.40±0.64),(23.28±1.30) MPa,经戊二醛交联后材料的抗压强度分别增大至(36.30±1.20),(51.60±2.08),(36.90±3.22) MPa。壳聚糖含量为10%,20%,30%壳聚糖/羟基磷灰石-庆大霉素交联后的吸水率与降解率均低于交联前。在体外缓释的第1天,30%壳聚糖/羟基磷灰石-庆大霉素的药物释放量为42.2%,材料经戊二醛交联处理后药物释放量降至33.6%,在随后的9 d,交联壳聚糖/羟基磷灰石-庆大霉素的总释放量均低于壳聚糖/羟基磷灰石-庆大霉素。表明戊二醛交联赋予了材料更好的生物稳定性,减缓了材料降解速率,显著改善了药物突释现象。中国组织工程研究杂志出版内容重点：生物材料;骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性;组织工程全文链接：     

Chitosan beads and granules for oral sustained delivery of nifedipine: in vitro studies.

Nifedipine was embedded in a chitosan matrix to develop a prolonged-release form. The in vitro release profiles of nifedipine from chitosan beads and microgranules were monitored by UV spectrophotometer. The studies were performed in a rotating shaker (100 rev min-1) in 0.1 M HCl buffer (pH 2.0) or 0.1 M phosphate buffer (pH 7.4). Comparison was made between drug-loaded microbeads and microgranules. The amount and percentage of drug release were much higher in HCl than in phosphate buffer, probably due to the salt formation of the matrix (chitosan hydrochloride) at acid pH. The release rate of nifedipine from chitosan matrix was slower for beads than granules. These findings suggest the possibility of modifying the formulations to obtain the desired controlled release of the drug in an oral sustained-delivery system.     

Electrospraying: a facile technique for synthesis of chitosan-based micro/nanospheres for drug delivery applications

Electrospraying is a novel technique for the generation of micro/nanospheres for biomedical applications. Apart from being a high yield technique; electrospraying has an added advantage of not making use of an external dispersion/emulsion phase which often involves ingredients that are undesirable for biomedical applications. In this study, we report the use of electrospraying for the synthesis of chitosan micro/nanospheres. The focus was to optimize the fabrication parameters involved in electrospraying for reproducible synthesis of chitosan based micro/nanospheres and to study their potential as delivery vehicles for bioactive agents. The influence of the following was studied (i) electrospraying voltage, (ii) needle gauge, (iii) concentration of chitosan solution, (iv) concentration of acetic acid solution, and (v) electrospraying distance. SEM analysis demonstrated that microspheres of less than 1 mum were obtained when chitosan concentration was 2% dissolved in 90% acetic acid. The working distance and needle gauge that yielded favorable results were 7 cm and 26 g, respectively. Average particle size of ampicillin loaded chitosan micro/nanospheres was 520 nm with zeta potential of +28.2 mV and encapsulation efficiency of 80.4%. The particles were characterized for drug release kinetics and results demonstrated an initial burst release followed by a sustained release over a period of 120 h. Further, antibacterial activity of drug loaded micro/nanospheres demonstrated that the encapsulated drug was in its active form postexposure to high voltage during electrospraying. This study indicates that electrospraying is a facile technique for the synthesis of chitosan micro/nanospheres for drug delivery applications.     

透明带3多肽-三甲基壳聚糖微球的制备及其在卵巢早衰中的应用

背景：透明带3多肽诱导口服耐受可预防及治疗自身免疫性卵巢早衰,但直接应用透明带3多肽治疗效果不十分理想,因此选择合适的药物载体系统成为进一步研究的基础。 目的：制备透明带3多肽-三甲基壳聚糖微球,观察其在卵巢早衰中的作用。 方法：采用离子交联法制备透明带3多肽-三甲基壳聚糖微球,观察微球形态,检测微球粒径、包封率、载药率及体外释放速度。分别以透明带3多肽-三甲基壳聚糖微球、磷酸盐缓冲液、透明带3多肽、三甲基壳聚糖灌胃治疗卵巢早衰小鼠。 结果与结论：透明带3多肽-三甲基壳聚糖微球形态较规则,平均粒径280.5 nm,包封率为69.20%,载药率为14.83%,随时间的延长,微球中透明带3多肽的体外释放率逐渐增加,无突释现象。透明带3多肽-三甲基壳聚糖微球组卵巢早衰小鼠外周血中抗透明带3多肽抗体阳性率明显低于其他3组(P < 0.05),表明透明带3多肽-三甲基壳聚糖微球治疗可明显减低卵巢早衰小鼠血清中抗透明带3多肽抗体阳性率。     

制备壳聚糖微球体外缓释TGF-β_1的研究

应用离子交联沉淀法制备壳聚糖-转化生长因子（TGF-β1）缓释微球,研究其体外缓释性能。采用离子交联沉淀法制备壳聚糖微球,以其包裹TGF-β1,制备具有缓释效能的壳聚糖-TGF-β1缓释微球。用扫描电镜、激光粒度分析仪、Elisa法等观察其表面形态,测定药物载药率、包封率、体外缓释效率等指标。结果表明：所得微球球形良好,表面光滑,粒径分布集中,平均粒径272nm。壳聚糖微球有较高的药物包封率,达80.60%。体外释放试验提示,TGF-β1初期存在突释现象,前24h释放达27%,但其后可从壳聚糖微球中稳定释放,7d累计释放达41%。离子交联沉淀法制备壳聚糖缓释微球方法简单易行,所得壳聚糖-TGF-β1微球具有良好的缓释效能,提示其在组织工程领域具有良好的应用前景。     

Drug-loaded biodegradable microspheres for image-guided combinatory epigenetic therapy in cells

We synthesize drug-loaded poly (lactic-co-glycolic acid) (PLGA) microspheres for image-guided combinatory epigenetic therapy in MCF-10A human mammary epithelial cells. LY294002 and Nile Red are encapsulated in microspheres for sustained drug release and fluorescence microscopic imaging. Drug-loaded microspheres target MCF-10A cells through a three-step binding process involving biotinylated antibody, streptavidin, and biotinylated microspheres. LY294002 loaded microspheres and 5-Aza-2-deoxycytidine are applied to MCF-10A cells for combinatory PI3K∕AKT inhibition and deoxyribonucleic acid (DNA) demethylation. Our study implies the technical potential of disease targeting and image-guided combinatory epigenetic therapy using drug-loaded multifunctional biodegradable PLGA microspheres.     

Controlled release of drugs from multi-component biomaterials

In order to control their release, drugs are encapsulated into systems which are expected to provide a certain site with a predetermined amount of drug over a well-defined period of time. Here we report on a multi-component drug delivery biomaterial that consists of a hydrogel matrix in which drug-loaded biodegradable microcarriers are dispersed, and whose potential applications could be found in the design of implantable devices with long-term activity, as required by contraceptive and hormone replacement treatments. The release profile of the drug can actually be tuned by the complex interplay of several release mechanisms, including the permeability and eventually the degradation rate of the microcarriers and the diffusion through the hydrogel. The hydrogel consisted of 2-hydroxyethyl methacrylate cross-linked by ethylene glycol dimethacrylate. The microcarriers were biodegradable poly-epsilon-caprolactone (PCL) microspheres in which active molecules, such as levonorgestrel (LNG), were encapsulated. The hydrogels were characterized by water swelling, thermal properties, LNG diffusion through drug-free and drug-depleted hydrogel membranes and LNG release from devices with drug dispersed in the hydrogel. The PCL microspheres were observed by scanning electron microscopy; their size distribution, LNG loading and release were also investigated. The hydrogel-microsphere assemblies were characterized in terms of the distribution of the microspheres within the hydrogel, water swelling and the release of the encapsulated molecules. The developed device, due to its composite structure, has the ability to combine several release mechanisms, leading to drug release obeying zero-order kinetics for most of the time.     

复合骨碎补总黄酮的丝素蛋白/壳聚糖支架在兔关节软骨损伤中的应用研究

目的 以丝素蛋白/壳聚糖支架为载体将骨碎补总黄酮应用于兔软骨损伤局部,观察修复效果,为临床提供实验数据。方法 制备丝素蛋白/壳聚糖支架、骨碎补总黄酮缓释微球与负载骨碎补总黄酮缓释微球的丝素蛋白/壳聚糖支架,扫描电子显微镜下观察支架形貌,同时检测该支架的体外缓释能力。24只新西兰大白兔随机分3组,利用电钻在股骨滑车部位构建直径3.5 mm、深1.5 mm的软骨损伤模型,空白组软骨缺损处不植入任何材料,对照组植入单纯的丝素蛋白/壳聚糖支架,实验组植入负载骨碎补总黄酮缓释微球的丝素蛋白/壳聚糖支架,术后12周、24周行标本大体与组织学观察,RT-PCR检测修复组织Sox-9、II型胶原与聚集蛋白聚糖mRNA的表达量,Western blot检测软骨缺损部位II型胶原蛋白表达,分析软骨修复效果。结果 丝素蛋白/壳聚糖支架具有良好的三维孔隙结构,孔洞之间相互联通;制备的载药微球表面较光滑,为较规则的圆球形;载药微球均匀分散于丝素蛋白/壳聚糖支架基质中。丝素蛋白/壳聚糖支架可在体外持续稳定地释放骨碎补总黄酮,实验组软骨损伤修复效果优于对照组,对应的ICRS评分与Wakitani组织学评分高于对照组...     

In vivo evaluation of a dexamethasone/PLGA microsphere system designed to suppress the inflammatory tissue response to implantable medical devices

The purpose of this research effort was to evaluate in vivo a newly developed dexamethasone/PLGA microsphere system designed to suppress the inflammatory tissue response to an implanted device, in this case a biosensor. The microspheres were prepared using an oil/water (O/W) emulsion technique. The microsphere system was composed of drug-loaded microspheres (including newly formulated and predegraded microspheres) and free dexamethasone. The combination of the drug and drug-loaded microspheres provided burst release of dexamethasone followed by continuous release from days 2-14. Continuous release to at least 30 days was achieved by mixing predegraded and newly formulated microspheres. The ability of our mixed microsphere system to control tissue reactions to an implant then was tested in vivo using cotton thread sutures to induce inflammation subcutaneously in Sprague-Dawley rats. Two different in vivo studies were performed, the first to find the dosage level of dexamethasone that effectively would suppress the acute inflammatory reaction and the second to show how effective the dexamethasone delivered by PLGA microspheres was in suppressing chronic inflammatory response to an implant. The first in vivo study showed that 0.1 to 0.8 mg of dexamethasone at the site minimized the acute inflammatory reaction. The second in vivo study showed that our mixed microsphere system suppressed the inflammatory response to an implanted suture for at least 1 month. This study has proven the viability of microsphere delivery of an anti-inflammatory to control the inflammatory reaction at an implant site.     

Biodegradable and complexed microspheres used for sustained delivery and activity protection of SOD

To develop a new protein delivery system for superoxide dismutase (SOD), biodegradable materials like poly(DL-lactide-co-glycolide) (PLGA), alginate, and chitosan were used for preparing PLGA microspheres and alginate-chitosan microspheres, which were used for encapsulating protein. Alginate-chitosan microspheres showed much higher entrapment efficiency (91.08% +/- 1.28%) than that of PLGA microspheres (36.42% +/- 1.81%). In vitro release study showed that SOD presented a sustained release character in the preparation of these biodegradable materials. After 15 days, 43.72% +/- 0.43% of protein was released from alginate-chitosan microspheres, while there was 62.96% +/- 3.95% of protein release from PLGA microspheres. However, alginate-chitosan demonstrated that it was a better material to control the burst release of protein from microspheres. Furthermore, SOD activity in microspheres was evaluated, and the results showed that microspheres protected the activity of protein to some extent. Finally, PLGA-alginate-chitosan complex microspheres were constructed and the release character in vitro demonstrated that this preparation could not only prolong the release of drug but also decrease the burst release.     

Drug-releasing scaffolds fabricated from drug-loaded microspheres.

Biodegradable scaffolds serve a central role in many strategies for engineering tissue replacements or in guiding tissue regeneration. Typically, these scaffolds function to create and maintain a space and to provide a support for cell adhesion. However, these scaffolds also can serve as vehicles for the delivery of bioactive factors (e.g., protein or DNA) in order to manipulate cellular processes within the scaffold microenvironment. This study presents a novel approach to fabricate tissue-engineering scaffolds capable of sustained drug delivery whereby drug-loaded microspheres are fabricated into structures with controlled porosity. A double-emulsion process was used to fabricate microspheres with encapsulated DNA that retained its integrity and was released from the microspheres within 24 h. These DNA-loaded microspheres subsequently were formed into a nonporous disk or an interconnected open-pore scaffold (>94% porosity) via a gas-foaming process. The disks and scaffolds exhibited sustained plasmid release for at least 21 days and had minimal burst during the initial phase of release. This approach of assembling drug-loaded microspheres into porous and nonporous structures may find great utility in the fabrication of synthetic matrices that direct tissue formation.     

Modified n-HA/PA66 scaffolds with chitosan coating for bone tissue engineering: cell stimulation and drug release

The dipping-drying procedure and cross-linking method were used to make drug-loaded chitosan (CS) coating on nano-hydroxyapatite/polyamide66 (nHA/PA66) composite porous scaffold, endowing the scaffold controlled drug release functionality. The prefabricated scaffold was immersed into an aqueous drug/CS solution in a vacuum condition and then crosslinked by vanillin. The structure, porosity, composition, compressive strength, swelling ratio, drug release and cytocompatibility of the pristine and coating scaffolds were investigated. After coating, the scaffold porosity and pore interconnection were slightly decreased. Cytocompatibility performance was observed through an in vitro experiment based on cell attachment and the MTT assay by MG63 cells which revealed positive cell viability and increasing proliferation over the 11-day period in vitro. The drug could effectively release from the coated scaffold in a controlled fashion and the release rate was sustained for a long period and highly dependent on coating swelling, suggesting the possibility of a controlled drug release. Our results demonstrate that the scaffold with drug-loaded crosslinked CS coating can be used as a simple technique to render the surfaces of synthetic scaffolds active, thus enabling them to be a promising high performance biomaterial in bone tissue engineering.     

壳聚糖抗肿瘤缓释药物的特征☆

背景：肿瘤严重危害人类的健康,应用有效的化学药物治疗具有重要的意义。 目的：分析壳聚糖载药缓释系统抗肿瘤的作用机制及治疗效果。 方法：分析壳聚糖抗肿瘤药物缓释材料释放药物机制以及抗肿瘤的作用机制,并分析壳聚糖及其衍生物负载多种抗肿瘤药物如阿霉素、表阿霉素、表柔比星、5-氟尿嘧啶、紫杉醇等的特征及抗肿瘤作用效果。 结果与结论：壳聚糖抗肿瘤缓释药物材料具有良好的药物缓释性能以及抗肿瘤性能,能够负载各种常用的抗肿瘤药物,可以控制药物释放速率,延长药物作用时间,维持有效的药物浓度,降低药物的毒副作用,并且对肿瘤及组织器官具有靶向性作用,明显提高了化疗药物对肿瘤的治疗作用。     

Synthesis, characterization, biodegradation, and drug delivery application of biodegradable lactic/glycolic acid polymers: Part III. Drug delivery application

Lactic/glycolic acid polymers (PLGA) are widely used for drug delivery systems. The microsphere formulation is the most interesting dosage form of the PLGA-based controlled release devices. In this study, the previously reported PLGA were used to prepare drug-containing microspheres. Progesterone was used as a model drug. The progesterone microspheres were prepared from PLGA having varied compositions and varied molecular weight. The microscopic characterization shows that the microspheres are spherical, nonaggregated particles. The progesterone-containing PLGA microspheres possess a Gaussian size distribution, having average size from 70-134 microm. A solvent extraction method was employed to prepare the microspheres. The microencapsulation method used in this study has high drug encapsulation efficiency. The progesterone release from the PLGA microspheres and the factors affecting the drug release were studied. The release of progesterone from the PLGA microspheres is affected by the properties of the polymer used. The drug release is more rapid from the microspheres prepared using the PLGA having higher fraction of glycolic acid moiety. The drug release from the microspheres composed of higher molecular weight PLGA is faster. The drug content in microspheres also has an effect on the drug release. Higher progesterone content in microspheres yields a quicker initial burst release of the drug.     

Sustained release of PEG-g-chitosan complexed DNA from poly(lactide-co-glycolide)

Chitosan and its derivatives have emerged as promising gene-delivery vehicles because of their capability to form polyplexes with plasmid DNA and enhance its transport across cellular membranes through endocytosis. Evidently, polyplexes of chitosan and DNA significantly improve transfection efficiency; however, these polyplexes are not capable of sustained DNA release and, thus, prolong gene transfer. In order to achieve prolonged delivery of DNA/chitosan polyplexes, we have formulated microspheres by physically combining poly(ethylene glycol)-grafted chitosan (PEG-g-CHN) with poly(lactide-co-glycolide) (PLGA) using a modified conventional in-emulsion solvent evaporation method. Electrophoretic analysis of materials released from these microspheres suggests the presence of PEG-g-CHN complexed DNA and these microspheres are capable of sustained release of DNA/PEG-g-CHN for at least 9 weeks. The rate of DNA release can be modulated by varying the amount of PEG-g-CHN. The release products from these microspheres are bioactive and show enhanced transfection in vitro over DNA released from conventional PLGA microspheres containing no PEG-g-CHN. In vivo experiments also show that these microspheres are capable of achieving gene transfer in a rat hind limb muscle model.     

负载木通皂苷D的纳米羟基磷灰石/壳聚糖支架修复骨缺损北大核心

背景:木通皂苷D具有促进成骨细胞的增殖与分化、提高成骨细胞活性与数量、促进基质钙化与骨痂生长等诸多作用,主要被用于治疗骨质疏松与促进骨折愈合,将其应用于骨缺损修复的研究较少见。目的:以纳米羟基磷灰石/壳聚糖支架为载体,将包裹木通皂苷D的缓释微球负载于其中,观察其骨缺损修复作用。方法:采用W/O/W方法制作包裹木通皂苷D的缓释微球,采用冷冻干燥方法制备负载包裹木通皂苷D缓释微球的纳米羟基磷灰石/壳聚糖支架(以下简称缓释支架)与单纯的纳米羟基磷灰石/壳聚糖支架(以下简称空白支架),检测缓释微球与缓释支架的体外释药能力。将小鼠来源前成骨细胞MC3T3-E1分别接种于两种支架上,以单独培养的细胞为对照,分析细胞的黏附、增殖与分化情况。在24只成年新西兰大白兔双侧桡骨中段制造1.5 cm的骨缺损,分别植入空白支架与缓释支架,术后4,12周时进行大体观察、Micro-CT扫描影像学检查及组织学观察。结果与结论:①包裹木通皂苷D的缓释微球与缓释支架均具有缓释作用,其中缓释支架的药物释放速率更加平稳、持久;②CCK-8实验显示,缓释支架上的细胞增殖速率快于空白支架、对照组(P<0.05);扫描电镜下可见,小鼠来源前成骨细胞MC3T3-E1覆盖在两组支架表面,其中缓释支架上的细胞数量要多于空白支架;③培养7,14 d时,缓释支架组的碱性磷酸酶活性、Runx2 mRNA表达高于空白支架组(P<0.05);培养第21天时,缓释支架组的骨桥蛋白、骨钙素蛋白表达量高于空白支架组(P<0.05);④影像学检查与组织学观察结果显示,术后4周时,缓释支架组材料周围可见大量的新生骨,其新生骨量明显多于空白支架组;术后12周时,缓释支架内部也可见大量的新生骨长入,空白支架内部仅见少量的新生骨长入,并且缓释支架组的材料残余明显少于空白支架组(P<0.05);⑤结果表明,负载木通皂苷D缓释微球的纳米羟基磷灰石/壳聚糖支架可提升体外成骨细胞的黏附、增殖与分化能力,提升纳米羟基磷灰石/壳聚糖支架的体内骨诱导能力。