残耳软骨细胞种植于聚羟基丁酸酯-聚羟基己酸酯共聚物形成组织工程软骨

目的探讨以聚羟基丁酸酯-聚羟基己酸酯(PHB-PHH)共聚物为细胞外支架、以残耳软骨作为种子细胞形成组织工程软骨的可能性。方法取先天性小耳畸形8例患者的残耳软骨，以胶原酶消化后种植于PHBPHH支架，体外培养1周后种植于8只裸鼠一侧背部皮下为实验组，另一侧只植入支架材料作为对照组。于4周、8周后取出标本，做大体观察及HE染色、Masson三色染色检查。结果4周时实验组镜下显示有新生软骨形成，但仍有部分支架材料残留；8周时实验组标本大体观察及HE染色、Masson三色染色检查新生软骨与人耳软骨相似，支架材料已完全吸收。对照组无软骨形成。结论以残耳软骨作为种子细胞，以PHB-PHH共聚物为细胞外支架可以形成组织工程软骨，新生软骨大体观察、组织学检查与人耳软骨相似。     

聚羟基丁酸酯与聚醚复合物作为软骨组织工程支架材料的实验研究

目前应用于组织工程研究的膜板支架材料有多种 ,但实际应用中任何材料单独应用都存在缺陷。本研究充分利用聚羟基丁酸酯 (ｐｏｌｙｈｙｄｒｏｘｙｂｕｔｙｒａｔｅ,ＰＨＢ)的可塑性和聚醚 (ｐｌｕｒｏｎｉｃＦ 12 7)对软骨细胞良好的粘固特性 ,将两种材料混合应用 ,观察此复合材料是否更适合组织工程化软骨的形成。1.材料与方法 :ＰＨＢ三维多孔泡沫材料由清华大学化工系研制 ,孔径2 5 0～ 35 0 μｍ ,孔隙率 95 %。将其裁剪成1 2ｃｍ× 1 1ｃｍ× 0 2ｃｍ方体后消毒 (酒精和紫外线 ) ,10 %ＰＬＹＳ(多聚赖氨酸 )包埋、晾干备用。将聚醚 (…     

聚羟基丁酸酯载体人工软骨体内培育的实验研究

目的 探讨聚羟基丁酸酶（ＰＨＢ）泡沫材料作为软骨细胞支架材料以及体内培育组织工程化人工软骨的可行性。方法 取４周龄雄性新西兰幼免关节软骨、胶原酶消化,将所获软骨细胞种植到ＰＨＢ泡沫材料上,体外培养１周后,将细胞－支架材料复合体移植到兔背背部下皮,以单纯植入聚羟基丁酸酯泡沫材料及接种软骨细胞组为对照组。分别于术后第４、８、１２周取材,进行大体观察及组织学、免疫组织化学观察。结果 软骨细胞＝支架材料复     

羟基聚磷酸钙钠与聚羟基丁酸酯膜联合修复骨缺损的实验研究

目的：评价羟基聚磷酸钙钠（Hydroxy poly-calcium nairium phosphate,HPA)与聚羟基丁酸酯膜(Polyhydroxybutyate,PHB）联合使用修复骨缺损的效果。方法：采用HPS充填狗股骨人工缺损区，同时覆盖PHB膜，与PHB膜覆盖及空白对照作比较，通过组织形态学及定量检测，考察1周，1、2、3月缺损修复情况。结果：HPA与PHB联合组，2月内骨密度及形态优于PHB组，3月后二组相当，均优于空白对照组。结论：HPA与PHB联合使用，可早期获得骨缺损修复强度与外形并举的效果。     

聚羟基丁酸酯可吸收缝线生物相容性的实验室研究

目的：研究聚羟基丁酸酯（Polyhydroxybutyrate,PHB）可吸收缝线的生物相容性。方法：①将成纤维细胞进行体外培养,传代后与PHB可吸收缝线复合培养,设与经聚氰基丙烯酸酯处理过的PHB可吸收缝线复合培养为对照组,观察细胞生长情况;②行扫描电镜检查观察细胞在丝线上的生长情况;③以MTT法检测复合培养后的细胞活性。结果：①实验组细胞生长良好,对照组细胞漂浮,收缩成团;②扫描电镜下见细胞在丝线上贴附生长良好;③MTT法检测实验组对细胞增殖无明显影响。结论：PHB可吸收缝线具有良好的生物相容性。     

羟基聚磷酸钙钠与聚羟基丁酸酯膜联合修复骨缺损的实验研究

目的 评价羟基聚磷酸钙钠(Hydroxy poly-calcium nairium phosphate,HPA)与聚羟基丁酸酯(Polyhydroxybutyrate,PHB)联合使用修复骨缺损的效果。方法 采用HPA充填狗股骨人工缺损区,同时覆盖PHB膜,与PHB膜覆盖及空白对照作比较,通过组织形态学及定量检测,考察1周,1、2、3月缺损修复情况。结果 HPA与PHB联合组,2月内骨密度及形态优于PHB组,3月后二组相当,均优于空白对照组。结论 HPA与PHB联合使用,可早期获得骨缺损修复强度与外形并举的效果。     

以聚β-羟基丁酸酯为支架的肾上腺细胞移植形态学研究

目的探讨以聚β-羟基丁酸酯(PHB)为支架的肾上腺细胞移植后的形态学变化。方法分离Wistar大鼠的肾上腺细胞,接种至改性的PHB上,然后移植至切除双侧肾上腺的大鼠的后肢肌间隙中,并设单纯双侧肾上腺切除的对照组及假手术组。观察术后第2、4、8周时移植局部的组织学变化,免疫组化染色观察移植局部CD3表达。结果组织学观察显示,随着时间的推移,移植物的组织反应逐渐变小,PHB逐渐降解,肾上腺细胞趋向组织化;超微结构观察显示,移植的肾上腺细胞保持着同正常肾上腺细胞相似的结构;免疫组织染色显示,移植的肾上腺细胞组织化后其CD3的表达与正常肾上腺组织相似(P>0.05)。结论以PHB为支架的肾上腺细胞移植,移植物在体内可存活,且呈现明显的组织化,移植的肾上腺细胞与正常肾上腺细胞的超微结构基本一致。     

以聚羟基丁酸已酯为支架材料的组织工程隆鼻法的实验研究

目的:研究以聚羟基丁酸已酯(PHB)为支架材料形成的组织工程软骨作为隆鼻填充材料的可行性。方法:实验组:取兔的耳软骨细胞,行体外培养扩增,达到一定数量后,再接种于PHB支架上,置于新西兰白兔的鼻背部骨膜下,从大体和组织学方面观察软骨形成情况;对照组:单纯以未接种软骨细胞的支架移植于兔的鼻背部骨膜下,于相同时间点进行检测。结果:实验组:植入4周后该复合物在兔鼻背下支架表层有软骨层形成,并可见软骨细胞突入至支架材料中间。8周后支架材料继续降解,软骨细胞及细胞基质生成明显,伴有少量血管及胶原长入,对照组无相应变化。结论:以聚羟基丁酸已酯(PHB)为支架材料的组织工程软骨可用于隆鼻的填充材料。     

聚羟基丁酸酯可吸收缝线灭菌方法的实验研究

目的：研究不同的灭菌方法对聚羟基丁酸酯（PHB）可吸收缝线的降解性和力学特性的影响。方法：将聚羟基丁酸酯（PHB）可吸收缝线进行酒精浸泡、环氧乙烷、紫外线照射三种方法进行灭菌处理后,用细菌培养检测灭菌效果,用粘度法测定聚合物分子量从而观察缝线的降解性,用拉伸试验测定缝线的力学特性。结果：①酒精浸泡灭菌组有10%检出细菌,环氧乙烷及紫外线灭菌组未检出细菌;②紫外线照射灭菌和酒精浸泡灭菌组可使缝线黏度下降并具有显著性差异,环氧乙烷对缝线黏度下降的影响没有显著性差异;③经酒精和紫外线灭菌后缝线的断裂强度降低明显,差异具有显著性,经环氧乙烷灭菌后缝线的断裂强度降低无显著性差异。经紫外线灭菌后缝线的断裂伸长率明显降低,差异具有显著性,经酒精和环氧乙烷灭菌后缝线的断裂伸长率无明显降低。结论：环氧乙烷灭菌对PHB可吸收缝线的降解性和力学特性影响较小,是PHB可吸收缝线较理想的灭菌方法。     

聚羟基丁酸酯复合骨髓基质细胞构建组织工程骨修复兔下颌骨缺损

目的：探讨以聚羟基丁酸酯（PHB）作为骨组织工程支架的可行性.方法：体外培养兔骨髓基质细胞（BMSCs）,使其向成骨细胞分化后接种于PHB支架上,植入兔下颌角骨缺损中,以单纯缺损、单纯植入材料、植入新鲜骨髓加材料作为对照.4、8、1 2、24周分别处死各组家兔2只,行大体标本、X线摄片、组织学及扫描电镜观察新骨形成情况.结果：实验组24周大部分材料被骨性组织取代,修复骨缺损效率较对照组高.结论：PHB可以作为组织工程材料中的一种来修复骨缺损.     

乳液冷冻干燥法制备明胶多孔支架及其性能研究

目的 采用乳液冷冻干燥法制备用于组织修复的明胶多孔支架材料,考察主要制备参数对支架材料性能的影响,为进一步制备满足细胞培养和临床应用的支架材料提供依据.方法 采用乳液冷冻干燥法,将明胶溶液与有机醇混合制成乳液,预冻,冷冻干燥得到支架材料,考察其性能.结果 制备过程中,乳液的固含量,预冻温度对支架材料的微观结构、表观密度、含水量和孔隙率有较大影响.结论 采用乳液冷冻干燥法可以成功的制备明胶多孔支架材料,通过控制反应条件和参数可以调整支架孔隙结构和性能,从而得到满足需要的支架材料.     

Osteogenesis Depending on Geometry of Porous Hydroxyapatite Scaffolds

The effect of the configuration of porous cylindrical hydroxyapatite (HA) scaffold and laminin preparation of the scaffold on bone formation was estimated. HA scaffolds with a hollow center of 2 or 4 mm in diameter and those without a hollow center were used. The scaffolds were immersed in laminin solution or in culture medium. Bone marrow cells were obtained from the femora of male Fischer 344 rats. Cell suspension was prepared at 1 x 10(6) cells/mL density. The cells were seeded into HA scaffolds. Each scaffold was implanted in the dorsal subcutis of rats for 4 weeks. Bone formation in scaffolds was observed histologically. The quantity of osteocalcin was measured immunochemically. Many pores containing bone were identified in the laminin-immersed HA scaffold with a hollow center measuring 4 mm in diameter than those without and those with a hollow center measuring 2 mm in diameter. A greater quantity of osteocalcin was detected in the HA scaffold with immersion in laminin than in that without immersion in laminin. However, the results of the immunochemical assay for osteocalcin showed that a hollow center in the scaffold did not contribute to bone formation compared to scaffolds without a hollow center. It is considered that laminin may act as an adhesive for effective cell attachment to the walls of the pores in an HA scaffold.     

Reconstruction of Saddle Nose Deformities Using Porous Polyethylene Implant

Various materials have been employed for nasal contour restoration. We used porous polyethylene implants in reconstruction of saddle nose deformity in 36 cases. Only one complication occurred in the 8–18 months follow-up period. No implant was removed. Both cosmetic and functional results were accepted as pleasing by the patients.     

两种组织工程化人工肋骨支架材料的制备及体内外降解实验研究

目的选择组织工程界常用的聚乳酸-羟基乙酸共聚物／羟基磷灰石（PLGA／HA）与多孔磷酸钙（CPC）两种支架材料,通过体内外降解实验筛选出更为符合组织工程化人工肋骨支架的材料。方法将PLGA／HA（PLGA／HA组）和CPC（CPC组）两种材料分别进行体外实验和动物体内实验,体外实验：将同等大小的PLGA／HA和CPC材料浸入0．9％NaCl溶液,并保持无菌,放入37℃温箱内,分别于2、4、8、12和24周时取出称重,比较两种材料在体外的降解速度差异;体内实验：将同等大小的PLGA／HA和CPC材料各2片分别植入20只新西兰大白兔脊柱两侧的皮下,于2、4、8、12和24周取出称重,各时间点分别处死4只大白兔;材料周围组织送组织学检查及扫描电子显微镜观察。结果薄层CT扫描（Micro—CT）和扫描电子显微镜观察结果,CPC组较PLGA／HA组具有较好的三维结构（1101．2228±0．6184mg／ccm vs．1072．5523±0．7442mg／ccm）及孔隙率（70．26％±0．45％ vs．72．82％±0．51％）;体外实验显示：两组材料在体外降解速度均较慢,至6个月时才有明显的降解,其中PLGA／HA组降解相对较多,降解了60％;体内实验显示：PLGA／HA组降解比体外更快,3个月时已基本降解完,比CPC组降解快,降解了96％;另外CPC组材料周围的炎症反应明显比PLGA／HA组轻,更适合细胞的生长和黏附。结论对再生时间较长的长段肋骨缺损,CPC比PLGA／HA更适合。     

Repair of Critical‐Sized Long Bone Defects Using Dipyridamole‐Augmented 3D‐Printed Bioactive Ceramic Scaffolds

There are over two million long bone defects treated in the United States annually, of which ~5% will not heal without significant surgical intervention. While autogenous grafting is the standard of care in simple defects, a customized scaffold for large defects in unlimited quantities is not available. Recently, a three‐dimensionally (3D)‐printed bioactive ceramic (3DPBC) scaffold has been successfully utilized in the of repair critical‐sized (CSD) long bone defects in vivo. In this study, 3DPBC scaffolds were augmented with dipyridamole (DIPY), an adenosine A2A receptor (A_2AR) indirect agonist, because of its known effect to enhance bone formation. CSD full thickness segmental defects (~11 mm × full thickness) defects were created in the radial diaphysis in New Zealand white rabbits (n = 24). A customized 3DPBC scaffold composed of β‐tricalcium phosphate was placed into the defect site. Groups included scaffolds that were collagen‐coated (COLL), or immersed in 10, 100, or 1,000 μM DIPY solution. Animals were euthanized 8 weeks post‐operatively and the radii/ulna‐scaffold complex retrieved en bloc, for micro‐CT, histological, and mechanical analysis. Bone growth was assessed exclusively within scaffold pores and evaluated by microCT and advanced reconstruction software. Biomechanical properties were evaluated utilizing nanoindentation to assess the newly regenerated bone for elastic modulus (E) and hardness (H). MicroCT reconstructions illustrated bone in‐growth throughout the scaffold, with an increase in bone volume dependent on the DIPY dosage. The histological evaluation did not indicate any adverse immune response while revealing progressive remodeling of bone. These customized biologic 3DPBC scaffolds have the potential of repairing and regenerating bone. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2499–2507, 2019     

金属与羟基磷灰石微孔表面股骨植入体与骨结合强度的比较研究

目的比较金属与羟基磷灰石(hydroxyapatite,HA)微孔表面股骨植入体与骨的结合强度.方法用等离子喷涂技术,分别喷涂HA微粒和钴铬钼合金微粒于不锈钢三棱针表面,配对植入15只成年家兔的股骨,饲养2个月作拔出试验,并比较手术当日与2个月后的X线改变.结果HA与钴铬钼合金涂层三棱针平均剪切强度分别为(0.98±0.12)MPa和(0.65±0.15)MPa,二者有显著性差异(P＜0.05).X线片HA涂层三棱针周围有较多的成骨反应,透明区较金属涂层三棱针要少而窄.结论植入兔的股骨短时间内,HA微孔表面植入体较金属微孔表面植入体更为稳定.     

Repair of Osteochondral Defects in a Rabbit Model Using a Porous Hydroxyapatite Collagen Composite Impregnated With Bone Morphogenetic Protein‐2

Articular cartilage has a limited capacity for spontaneous repair, and an effective method to repair damaged articular cartilage has not yet been established. The purpose of this study was to evaluate the effect of transplantation of porous hydroxyapatite collagen (HAp/Col) impregnated with bone morphogenetic protein‐2 (BMP‐2). To evaluate the characteristics of porous HAp/Col as a drug delivery carrier of recombinant human BMP‐2 (rhBMP‐2), the rhBMP‐2 adsorption capacity and release kinetics of porous HAp/Col were analyzed. Porous HAp/Col impregnated with different amounts of rhBMP‐2 (0, 5, and 25 μg) was implanted into osteochondral defects generated in the patellar groove of Japanese white rabbits to evaluate the effect on osteochondral defect regeneration. At 3, 6, 12, and 24 weeks after operation, samples were harvested and subjected to micro‐computed tomography analysis and histological evaluation of articular cartilage and subchondral bone repair. The adsorption capacity was 329.4 μg of rhBMP‐2 per cm³ of porous HAp/Col. Although 36% of rhBMP‐2 was released within 24 h, more than 50% of the rhBMP‐2 was retained in the porous HAp/Col through the course of the experiment. Defects treated with 5 μg of rhBMP‐2 showed the most extensive subchondral bone repair and the highest histological regeneration score, and differences against the untreated defect group were significant. The histological regeneration score of defects treated with 25 μg of rhBMP‐2 increased up to 6 weeks after implantation, but then decreased. Porous HAp/Col, therefore, is an appropriate carrier for rhBMP‐2. Implantation of porous HAp/Col impregnated with rhBMP‐2 is effective for rigid subchondral bone repair, which is important for the repair of the smooth articular surface.     

单层组织工程纤维环取向纳米纤维支架的构建和初步验证

目的 使用静电纺丝技术构建取向纳米纤维支架,观测其结构,测试其力学特点,观察支架与细胞之间的相互作用,探讨其作为纤维环组织工程支架的可能性.方法 使用高速滚轴作为收集装置制备聚左旋乳酸聚己内酯[poly（lactic acid-co-caprolactone,P（LIA-CL）]取向纳米纤维支架.以P（LLA-CL）无规纳米纤维支架作为对照,观察取向纳米纤维支架的纤维直径、角度分布、孔径等表观指标;测试其拉伸力学行为;体外接种骨髓间充质干细胞观察其体外增殖和生长形态情况.结果 P（LLA-CL）取向纳米纤维支架纤维取向性良好,平行于纤维方向上的拉伸力学性能得到明显增强.接种细胞后,支架可促进细胞增殖,并诱导细胞沿着纤维方向拉伸并定向排列.结论 取向纳米纤维支架的力学强度在取向方向上得到了明显增强,具有和单层纤维环相似的各向异性力学特点,并可以诱导细胞沿纤维方向定向排列,细胞分布与纤维环细胞排列方式相似,可用于纤维环组织工程研究.