羟基丁酸-羟基辛酸聚合物-胶原骨软骨支架的制备及应用

背景：关节软骨修复的关键是软骨和软骨下骨的整体修复,然而目前尚缺乏理想的一体化支架。 目的：制备聚羟基丁酸-羟基辛酸-胶原一体化支架,并分析其基本生物学特性。 方法：以聚羟基丁酸-羟基辛酸、Ⅰ型胶原为材料,通过溶剂浇铸-颗粒沥滤法制备聚羟基丁酸-羟基辛酸-胶原一体化支架,观察支架超微结构,支架孔径及孔与孔的连通情况;液体置换法测定支架孔隙率。将乳兔骨髓间充质干细胞接种于聚羟基丁酸-羟基辛酸-胶原一体化支架上,扫描电镜观察细胞在支架上的黏附状态,MTT法测定细胞在支架上的生长曲线。 结果与结论：一体化支架呈疏松多孔结构,软骨层孔径80-100 μm,骨层孔径200-220 μm,孔隙率(80.0±2.3)%。骨髓间充质干细胞在支架上黏附状态良好,增殖迅速。说明聚羟基丁酸-羟基辛酸-胶原骨软骨一体化支架具备适宜的孔隙结构和良好的生物亲和性。     

羟基丁酸-羟基辛酸共聚体/胶原骨软骨一体化支架修复膝关节软骨损伤

BACKGROUND: Due to the complex physiological characteristics of the osteochondral tissue, the clinical repair of knee cartilage injury often has dissatisfied outcomes. Tissue engineering methods and tools provide a new idea for osteochondral repair. OBJECTIVE: To observe the effect of poly(hydroxybutyrate-co-hydroxyoctanoate/collagen) osteochondral tissue-engineered scaffold on the repair of articular cartilage injury in a rabbit. METHODS: The poly(hydroxybutyrate-co-hydroxyoctanoate/collagen) osteochondral tissue-engineered scaffold was prepared by solvent casting/particle leaching method. Then, seed cells were isolated and cultured on the scaffold. Twenty-four healthy New Zealand white rabbits, 4 weeks of age, were used for the study. Under balanced anesthesia, an articular cartilage defect (4.5 mm in diameter, 5 mm in depth) was created on the rabbit’s femoral condyle using a bone drill. After modeling, rabbits were randomized into three groups and given direct suture in blank group, pure scaffold implantation in control group and implantation of the scaffold-cell complex in experimental group. Femoral condyle of each rabbit was taken out for gross and histological observations at 8, 20 weeks after surgery. RESULTS AND CONCLUSION: At 8 weeks after surgery, transparent film-covered defects and small/irregular cells were found in the experimental group; the defects were filled with fibrous tissues in the control group; while there was no repair in the blank group. Until the 20th week, the defects were covered with hyaline cartilage-like tissues, accompanied by regular cell arrangement in the experimental group; in the control group, the defects were covered with white membranous tissues, and many chondrocytes were found at the basement and edge; in the blank group, some newborn tissues were visible at the defect region. These findings suggest that the poly (hydroxybutyrate-co- hydroxyoctanoate/collagen) osteochondral tissue-engineered scaffold carrying seed cells contributes to articular cartilage repair.     

羟基丁酸-羟基辛酸一体化骨软骨支架的体外血管化

背景：前期实验构建的羟基丁酸-羟基辛酸共聚体一体化骨软骨支架具备良好的生物相容性、生物可降解性,并且降解产物无毒性。 目的：将兔肾微血管内皮细胞与羟基丁酸-羟基辛酸一体化骨软骨支架复合培养,观察支架骨层血管化效果。 方法：运用溶剂浇铸-颗粒沥滤法,制备具有骨层/骨与软骨界面层/软骨层3层结构的羟基丁酸-羟基辛酸一体化骨软骨支架。将传代培养至第3 代的兔肾微血管内皮细胞,接种到一体化骨软骨支架骨层支架上,MTT法检测细胞在支架上的增殖活性,10 d后苏木精-伊红染色及电镜观察细胞在支架内的生长状况。 结果与结论：一体化骨软骨支架外观具备明显的3层结构,各层之间连接紧密,骨层疏松多孔,各层支架孔隙均匀且相通,一体化支架孔隙率为78%。兔肾微血管内皮细胞在支架上分裂增殖良好,复合培养10 d后,细胞在骨层支架内呈立体生长,中间界面层内未发现细胞,苏木精-伊红染色可见细胞黏附生长于骨层支架孔隙间,细胞依附支架的多孔结构生长,形成管腔样结构,但细胞并未长入中间界面层。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：     

羟基丁酸与羟基辛酸共聚物一体化骨软骨组织工程支架的制备及性能*◆

背景：单层支架难以满足关节软骨损伤修复的要求,现提出骨软骨共同修复的一体化支架,以弥补了单一支架的部分缺陷。 目的：以羟基丁酸与羟基辛酸共聚物为基础材料,羟基磷灰石等为复合材料研制一体化骨软骨组织工程支架,测试该支架的物理特性和细胞黏附性。 方法：采用溶剂浇铸/颗粒沥滤法,以支架孔径、孔隙率、力学强度和细胞黏附生长率为检测指标,以羟基丁酸与羟基辛酸共聚物为连续相,通过改变致孔剂NaCl粒径和羟基磷灰石材料配比制备不同形态结构、力学强度和生物学功能的三层一体化骨软骨组织工程支架。 结果与结论：致孔剂与支架材料的最佳质量配比分别为软骨层4.5/1,过渡层2.5/1,硬骨层3.5/1。扫描电镜观察显示支架的三层结构明显不同且紧密结合,其软骨层、过渡层、硬骨层的孔径分别为150~250 μm,≤60 μm,150~450 μm;孔隙率检测结果依次为84%,60%,75%;力学强度测定依次为2.93,6.43,4.30 MPa;支架对骨髓间充质干细胞无毒性,细胞黏附与生长状态良好。结果表明该一体化骨软骨组织工程支架具有仿生学特性,符合骨软骨组织工程支架的基本条件。关键词：羟基丁酸;羟基辛酸;共聚物;一体化支架;关节软骨缺损;骨软骨组织工程 缩略语注释：PHBHO：poly(hydroxybutyrate-co-hydroxyoctanoate),羟基丁酸与羟基辛酸共聚体;HA：hydroxyapatite,羟基磷灰石 doi:10.3969/j.issn.1673-8225.2012.16.004     

羟基丁酸-羟基辛酸共聚物/脱细胞软骨基质复合支架的制备与体外降解*

背景：羟基丁酸-羟基辛酸共聚物具有高生物相容性和降解性,但单一材料无法满足组织工程支架的要求。脱细胞软骨基质是制备复合支架的常用材料,具有良好的生物相容性和无抗原性等优点。 目的：将羟基丁酸-羟基辛酸共聚物、脱细胞软骨基质以不同比例混合制备复合支架,观察其体外降解速率。 方法：取新鲜猪关节软骨,用含有双抗的PBS液浸泡,放入含有苯甲基磺酰氟的tri-HCl缓冲液,加入DNase酶和RNase酶,用D-Hank’s液冲洗等步骤制备脱细胞软骨基质。采用溶剂浇注-颗粒沥滤的方法与羟基丁酸-羟基辛酸共聚物按不同浓度混合,制备出不同比例的复合支架。 结果与结论：脱细胞软骨基质的比例不同,复合材料的完全降解时间也不同,8%含量的脱细胞软骨基质最符合软骨组织工程支架的要求。      

羟基磷灰石/胶原-聚乳酸三维多孔框架材料的制备成形及分析

目的：合成新型的复合生物材料框架作为骨组织工程研究的细胞外基质材料。方法：本研究采用材料学自组装技术的原理，以Ⅰ型胶原蛋白为分子模板，引导钙磷盐在液相中的矿化，制备具有天然骨基质层状结构的羟基磷灰石／胶原复合材料，并以热致分相法制备了羟基磷灰石／胶原-聚乳酸复合三维多孔框架。结果：羟基磷灰石／胶原复合材料具有与天然骨基质相似的成分与结构，加入聚乳酸制备成三维多孔框架，孔隙直径界于50um-300um。结论：羟基磷灰石／胶原-聚乳酸复合三维多孔框架可能作为骨组织工程良好的细胞外基质材料。     

细胞外基质来源支架在软骨组织工程中的应用

BACKGROUND: At present, a variety of extracellular matrix-derived scaffolds have been successfully applied for cartilage tissue engineering in experiment and clinical practice. OBJECTIVE: To summarize the application and research status of extracellular matrix-derived scaffolds in cartilage tissue engineering. METHODS: A computer-based online search in PubMed, CNKI, CqVip and WanFang databases was performed using the keywords of “tissue engineering, cartilage, extracellular matrix, scaffolds” in English and Chinese, respectively. A total of 1 140 literatures were retrieved, and finally 65 eligible literatures were included. RESULTS AND CONCLUSION: In terms of the components, extracellular matrix-derived scaffolds are divided into monomeric natural polymers, mixed natural polymers, natural polymers compositing with synthetic polymers as well as acellular extracellular matrix-derived materials. Extracellular matrix-derived scaffolds hold good biocompatibility and degradability, and can promote proliferation and differentiation of choncrodytes; therefore, they as good bionic scaffolds have been applied for cartilage tissue engineering in clinical practice, However, poor mechanical properties and difficulty to molding should never be ignored. Further research should focus on improving the preparation technology by combining synthetic materials with extracellular matrix-derived scaffolds for cartilage tissue engineering.      

以羟基丁酸酯与羟基已酸酯共聚物为支架材料构建组织工程喉形态软骨

背景：再生预定形态组织工程软骨的研究为喉软骨病损的修复与重建提供了新思路与新方法。然而,由于喉软骨形态、部位与功能的特殊性,迄今在此领域软骨组织工程研究并未呈现出其应有的优势。目的：探讨带蒂肌筋膜组织瓣构建组织工程喉支架形态软骨方法,为肌筋膜瓣复合组织工程化软骨修复重建喉软骨支架功能提供实验依据。方法：采用溶剂浇铸、模压成形和颗粒滤沥方法制备喉支架形态聚羟基丁酸酯与聚羟基己酸酯共聚物聚羟基丁酸酯与聚羟基己酸酯共聚物聚羟基丁酸酯与聚羟基己酸酯共聚物生物材料塑形物,接种软骨细胞形成细胞-聚羟基丁酸酯与聚羟基己酸酯共聚物复合物,体外共同培养1周后用于体内植入。将新西兰白兔脊背部一侧骶棘肌及其筋膜制备肌筋膜组织瓣,采用筋膜衬里方法充填与包裹软骨细胞聚羟基丁酸酯与聚羟基己酸酯共聚物喉支架形态复合物,原位植入。将单纯聚羟基丁酸酯与聚羟基己酸酯共聚物喉支架体内植入的兔作为对照组。分别于术后6,12和18周取材,行大体形态观察、组织学和免疫组化检测评估喉支架形态组织工程化软骨成形与再生情况。结果与结论：制备的聚羟基丁酸酯与聚羟基己酸酯共聚物多孔生物材料塑形物呈中空半面喇叭状,形似喉支架形态,乙醇静态容积测定孔隙率>90%。筋膜衬里的带蒂肌筋膜组织瓣血运丰富,可有效充填与包裹喉支架形态塑形物。不同时间点均获取形态维持良好的喉支架形态组织工程软骨,组织学和免疫组化检测均证实体内植入6周即可形成软骨组织,12周及18软骨组织进一步成熟,而对照组体内植入未检测到软骨组织。结果证实,带蒂肌筋膜组织瓣可保障血运,采用筋膜衬里的肌筋膜组织瓣充填与包裹方法可构建喉支架形态组织工程软骨。 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     

纳米羟基磷灰石/聚羟基丁酸酯骨组织工程支架的制备及性能表征

背景：高分子聚合物聚羟基丁酸酯具有优良的生物相容性、生物降解性及压电性,但也存在脆性大、亲水性较差等不足。 目的：制备不同组成比例的电纺纳米羟基磷灰石/聚羟基丁酸酯纤维支架材料,分析其结构及性能。 方法：通过气流-高压静电纺丝技术制备纳米羟基磷灰石质量百分比分别为0、10%、20%、30%的电纺纳米羟基磷灰石/聚羟基丁酸酯纤维支架,检测支架材料的微观结构、基团组成、晶相分布、热学性能及表面润湿性。 结果与结论：扫描电镜观察显示,随着纳米羟基磷灰石含量的增大,越来越多的纳米羟基磷灰石颗粒分布于复合纤维表面且分布趋于均匀,到含量达到30%时,纳米羟基磷灰石已基本布满纤维表面,纤维表面的粗糙度也随之增加;差示扫描量热法及X射线衍射结果表明,纳米羟基磷灰石的加入可以降低复合纤维中聚羟基丁酸酯的结晶度及结晶规整程度,且纳米羟基磷灰石含量越大效果越明显;随着纳米羟基磷灰石含量的增大,复合支架表面的接触角逐渐降低,亲水性有所提高。表明将纳米羟基磷灰石与聚羟基丁酸酯复合进行电纺可以有效提高材料的表面润湿性及结晶度,改善材料的亲水性及脆性,且纳米羟基磷灰石含量越高作用越明显。 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     

胶原-纳米羟基磷灰石复合支架的细胞相容性

背景：观察成骨细胞在生物材料上的形态、增殖和分化等项目,可评估生物支架材料的生物相容性。 目的：观察复合支架材料纳米羟基磷灰石/胶原对成骨细胞增殖、分化的影响。 方法：取新生24 h内Wistar大鼠的颅盖骨,采用改良胶原酶消化法进行成骨细胞原代培养,取第3代细胞与纳米羟基磷灰石/胶原支架或普通羟基磷灰石材料体外复合培养。培养3,6,9 d后,观察材料周边的细胞形态及支架材料对细胞分化、增殖的影响。 结果与结论：纳米羟基磷灰石/胶原材料较普通的羟基磷灰石材料更有利于成骨细胞的黏附、生长、分化、增殖,证实其生物相容性更好,有望成为一种新型的骨组织工程支架材料。     

Jellyfish collagen scaffolds for cartilage tissue engineering

Porous scaffolds were engineered from refibrillized collagen of the jellyfish Rhopilema esculentum for potential application in cartilage regeneration. The influence of collagen concentration, salinity and temperature on fibril formation was evaluated by turbidity measurements and quantification of fibrillized collagen. The formation of collagen fibrils with a typical banding pattern was confirmed by atomic force microscopy and transmission electron microscopy analysis. Porous scaffolds from jellyfish collagen, refibrillized under optimized conditions, were fabricated by freeze-drying and subsequent chemical cross-linking. Scaffolds possessed an open porosity of 98.2%. The samples were stable under cyclic compression and displayed an elastic behavior. Cytotoxicity tests with human mesenchymal stem cells (hMSCs) did not reveal any cytotoxic effects of the material. Chondrogenic markers SOX9, collagen II and aggrecan were upregulated in direct cultures of hMSCs upon chondrogenic stimulation. The formation of typical extracellular matrix components was further confirmed by quantification of sulfated glycosaminoglycans.     

以交联透明质酸钠为支架体外构建组织工程软骨

背景：采用组织工程技术再生和重建软骨是目前修复软骨组织缺损效果最好、最有应用前景的方法。 目的：以体外培养的软骨细胞和交联透明质酸钠为支架材料,开发一套体外构建组织工程软骨的完整方案。 方法：分离新西兰兔膝关节软骨细胞,制成细胞悬液滴加于交联透明质酸钠支架上,体外复合培养21 d,提取RNA进行RT-PCR检测,制备冰冻切片进行显微观察和免疫组织化学观察。 结果与结论：软骨细胞接种于交联透明质酸钠支架材料后,可贴附于支架上生长,并且大量细胞聚集成团,在支架材料的纤维间隙中生长或呈单层细胞附着于支架材料纤维。细胞-支架复合物表达软骨组织特异性蛋白聚糖基因和Ⅱ型胶原α1基因,以及软骨组织特异性蛋白Ⅱ型胶原蛋白,可维持软骨细胞表型。表明培养的细胞-支架复合物在体外培养可形成软骨细胞外基质,有望获得组织工程软骨组织。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：     

Novel collagen scaffolds prepared by using unnatural D-amino acids assisted EDC/NHS crosslinking

This work discusses the preparation and characterization of novel collagen scaffolds by using unnatural D-amino acids (Coll-D-AAs)-assisted 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC)/N-hydroxyl succinimide(NHS)-initiated crosslinking. The mechanical strength, hydrothermal and structural stability, resistance to biodegradation and the biocompatibility of Coll-D-AAs matrices were investigated. The results from Thermo mechanical analysis, Differential scanning calorimetric analysis and Thermo gravimetric analysis of the Coll-D-AAs matrices indicate a significant increase in the tensile strength (TS, 180?±?3), % elongation (% E, 80?±?9), elastic modulus (E, 170?±?4) denaturation temperature (T _d, 108?±?4) and a significant decrease in decomposition rate (T _g, 64?±?6). Scanning electron microscopic and Atomic force microscopic analyses revealed a well-ordered with properly oriented and well-aligned structure of the Coll-D-AAs matrices. FT-IR results suggest that the incorporation of D-AAs favours the molecular stability of collagen matrix. The D-AAs stabilizing the collagen matrices against degradation by collagenase would have been brought about by protecting the active sites in collagen. The Coll-D-AAs matrices have good biocompatibility when compared with native collagen matrix. Molecular docking studies also indicate better understanding of bonding pattern of collagen with D-AAs. These Coll-D-AAs matrices have been produced in high mechanical strength, thermally and biologically stable, and highly biocompatible forms that can be further manipulated into the functional matrix suitable in designing scaffolds for tissue engineering and regenerative medical applications.     

Development of arginine-glycine-aspartate-immobilized 3D printed poly(propylene fumarate) scaffolds for cartilage tissue engineering

Abstract

Poly(propylene fumarate) (PPF) has known to be a good candidate material for cartilage tissue regeneration because of its excellent mechanical properties during its degradation processes. Here, we describe the potential application of PPF-based materials as 3D printing bioinks to create macroporous cell scaffolds using micro-stereolithography. To improve cell-matrix interaction of seeded human chondrocytes within the PPF-based 3D scaffolds, we immobilized arginine-glycine-aspartate (RGD) peptide onto the PPF scaffolds. We also evaluated various cellular behaviors of the seeded chondrocytes using MTS assay, microscopic and histological analyses. The results indicated that PPF-based biocompatible scaffolds with immobilized RGD peptide could effectively support initial adhesion and proliferation of human chondrocytes. Such a 3D bio-printable scaffold can offer an opportunity to promote cartilage tissue regeneration.     

聚乙烯醇及其复合材料在组织工程支架中的应用

背景：聚乙烯醇是具有良好生物相容性和生物降解特性的聚合物,因其水溶性、成膜性、乳化性、胶黏性,而且无味无毒,被广泛用于临床领域。目的：综述聚乙烯醇及其复合材料在骨、软骨、皮肤、血管等组织工程支架中的应用。方法：由第一作者检索2000年1月至2011年12月中国知网数据库、1980年1月至2012年12月Pubmed数据库及 Elsevier数据库中,有关聚乙烯醇及其复合材料在骨、软骨、皮肤、血管等组织工程支架中应用的文章,中文关键词为 “聚乙烯醇,复合材料,组织工程支架”,英文关键词为 “Poly (vinyl alcohol),composite material, tissue engineering scaffold”。结果与结论：虽然聚乙烯醇及其复合材料还存在强度不够高、植入后有并发症等缺点,但这类材料具有良好的生物相容性和生物可降解特性,在组织工程中的应用从实验室到临床前研究都有很大的进展。对于其修复的长期效果还需要进一步深入研究。通过对材料表面进行修饰,改善细胞与支架材料的相互作用;通过模拟细胞生长微环境,制备仿生材料,提高材料的亲水性、对细胞的黏附性,促进细胞的分化增殖;构建具有可控三维多孔结构的支架,并赋予其控制释放细胞生长因子等功能,更好地仿生天然细胞外基质的结构和功能;制备出降解速度与机械强度能够完全适应组织再生需要的支架,研制复合、仿生材料是今后支架材料研究的主要方向。     

Crosslinked poly(epsilon-caprolactone/D,L-lactide)/bioactive glass composite scaffolds for bone tissue engineering

A series of elastic polymer and composite scaffolds for bone tissue engineering applications were designed. Two crosslinked copolymer matrices with 90/10 and 30/70 mol % of epsilon-caprolactone (CL) and D,L-lactide (DLLA) were prepared with porosities from 45 to 85 vol % and their mechanical and degradation properties were tested. Corresponding composite scaffolds with 20-50 wt % of particulate bioactive glass (BAG) were also characterized. Compressive modulus of polymer scaffolds ranged from 190+/-10 to 900+/-90 kPa. Lactide rich scaffolds absorbed up to 290 wt % of water in 4 weeks and mainly lost their mechanical properties. Caprolactone rich scaffolds absorbed no more than 110 wt % of water in 12 weeks and kept their mechanical integrity. Polymer and composite scaffolds prepared with P(CL/DLLA 90/10) matrix and 60 vol % porosity were further analyzed in simulated body fluid and in osteoblast culture. Cell growth was compromised inside the 2 mm thick three-dimensional scaffold specimens as a static culture model was used. However, composite scaffolds with BAG showed increased osteoblast adhesion and mineralization when compared to neat polymer scaffolds.     

无纺网与多孔海绵状材料作为软骨组织工程支架的适用性及体内降解性

背景：聚羟基乙酸无纺网与聚羟基丁酸酯-聚羟基己酸酯共聚物多孔海绵具有良好的塑形适应性、生物降解性与生物相容性。 目的：观察聚羟基乙酸无纺网与聚羟基丁酸酯-聚羟基己酸酯共聚物多孔海绵作为软骨组织工程支架的适用性及体内降解性。 方法：分别制备乳兔软骨细胞-聚羟基乙酸无纺网复合物、乳兔软骨细胞-聚羟基丁酸酯-聚羟基己酸酯共聚物多孔海绵复合物。在实验组成年兔两侧背部皮下分别植入制备的两种复合物,在对照组成年兔两侧背部皮下分别植入聚羟基乙酸无纺网与聚羟基丁酸酯-聚羟基己酸酯共聚物。 结果与结论：组织学观察显示,以聚羟基乙酸无纺网获取的组织工程软骨,植入4 周时软骨细胞较小,软骨内有较多聚羟基乙酸纤维残留,8周时软骨细胞较成熟,包埋在陷窝内,聚羟基乙酸纤维消失,12周时软骨细胞成熟,基质分泌丰富,无聚羟基乙酸存留;以聚羟基丁酸酯-聚羟基己酸酯共聚物多孔海绵获取的组织工程软骨,植入4周时软骨细胞不成熟,软骨基质内似“杂质”样材料残留物较多,8周时软骨细胞较成熟,软骨基质内仍可见材料残留,12周时软骨基质材料残留基本消失。两组组织工程软骨特殊染色与免疫组织化学检测均显示再生软骨胶原与基质黏多糖生成良好,软骨中均检测出Ⅱ型胶原。表明两种材料作为软骨组织工程支架具有良好的适用性,其降解时间均达到组织工程软骨构建的要求。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：