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1.
Articular cartilage defects do not regenerate. Transplantation of autologous articular chondrocytes, which is clinically being performed since several decades, laid the foundation for the transplantation of genetically modified cells, which may serve the dual role of providing a cell population capable of chondrogenesis and an additional stimulus for targeted articular cartilage repair. Experimental data generated so far have shown that genetically modified articular chondrocytes and mesenchymal stem cells (MSC) allow for sustained transgene expression when transplanted into articular cartilage defects in vivo. Overexpression of therapeutic factors enhances the structural features of the cartilaginous repair tissue. Combined overexpression of genes with complementary mechanisms of action is also feasible, holding promises for further enhancement of articular cartilage repair. Significant benefits have been also observed in preclinical animal models that are, in principle, more appropriate to the clinical situation. Finally, there is convincing proof of concept based on a phase I clinical gene therapy study in which transduced fibroblasts were injected into the metacarpophalangeal joints of patients without adverse events. To realize the full clinical potential of this approach, issues that need to be addressed include its safety, the choice of the ideal gene vector system allowing for a long-term transgene expression, the identification of the optimal therapeutic gene(s), the transplantation without or with supportive biomaterials, and the establishment of the optimal dose of modified cells. As safe techniques for generating genetically engineered articular chondrocytes and MSCs are available, they may eventually represent new avenues for improved cell-based therapies for articular cartilage repair. This, in turn, may provide an important step toward the unanswered question of articular cartilage regeneration. 相似文献
2.
目的观察分析大转子骨瓣表面骨膜及腱膜等纤维结缔组织向关节软骨转化的规律及胰岛素样生长因子-1(IGF-1)在软骨化生过程中的作用。方法制备液氮冷冻双侧股骨头缺血性坏死(Osteonecrosis of femoral head,ONFH)的动物模型。左侧股骨头造模后即缝合关节囊,右侧股骨头根据分组不同采用不同的处理方式:A组(骨瓣治疗组):带血管蒂大转子骨瓣进行股骨头再造;B组(骨瓣加Ad-IGF-1基因治疗治疗组):带血管蒂大转子骨瓣股骨头再造,关节内注入表达IGF-1的腺病毒载体(Ad-IGF-1);两组动物分别于3,6,12,18,24周每批4只处死,对骨瓣进行大体观察,组织病理学观察,免疫组化检测。结果所有动物左侧冷冻区组织坏死,纤维状物覆盖,碎片样组织修复。组织病理切片及免疫组化证实A组右侧骨瓣区自6周出现透明软骨细胞,B组右侧骨瓣区自3周出现透明软骨细胞,B组较A组修复效果好。结果经统计学处理有统计学意义。结论大转子表面的骨膜及腱膜能够向关节软骨化生,IGF-1对大转子表面的骨膜及腱膜向关节软骨化生有促进作用,为ONFH的外科治疗及生长因子的应用提供基础。 相似文献
4.
Full-thickness defects of articular cartilage were repaired by implantation of porous polymer implants in rabbits and dogs. The quality of the repair tissue was determined by collagen typing with antibodies. Implants with varying pore sizes and chemical composition were used. The effect of loading and motion was determined by inserting implants higher than, level with and lower than the surrounding cartilage. It appeared that healing took place by formation of fibrocartilaginous repair tissue containing both type I and type II collagen. Hyaline cartilage was observed in a minority of the rabbits used but not in the dog. Fibrocartilage formation in the dog was simulated by implantation of a porous polymer. Chemical composition of the polymer did not alter the results, neither did loading of the implant. It is concluded that the formation of fibrocartilaginous repair cartilage is stimulated by implantation of a porous polymer. This tissue seemed to function adequately in the dog but did show signs of degeneration in the rabbit. 相似文献
5.
The repair of articular cartilage defects remains a significant challenge in orthopedic medicine. Hydrogels, three-dimensional polymer networks swollen in water, offer a unique opportunity to generate a functional cartilage substitute. Hydrogels can exhibit similar mechanical, swelling, and lubricating behavior to articular cartilage, and promote the chondrogenic phenotype by encapsulated cells. Hydrogels have been prepared from naturally derived and synthetic polymers, as cell-free implants and as tissue engineering scaffolds, and with controlled degradation profiles and release of stimulatory growth factors. Using hydrogels, cartilage tissue has been engineered in vitro that has similar mechanical properties to native cartilage. This review summarizes the advancements that have been made in determining the potential of hydrogels to replace damaged cartilage or support new tissue formation as a function of specific design parameters, such as the type of polymer, degradation profile, mechanical properties and loading regimen, source of cells, cell-seeding density, controlled release of growth factors, and strategies to cause integration with surrounding tissue. Some key challenges for clinical translation remain, including limited information on the mechanical properties of hydrogel implants or engineered tissue that are necessary to restore joint function, and the lack of emphasis on the ability of an implant to integrate in a stable way with the surrounding tissue. Future studies should address the factors that affect these issues, while using clinically relevant cell sources and rigorous models of repair. 相似文献
6.
In the present study, NGF, BNDF from the neurotrophin family and IGF-1 were covalently immobilized on gelatin-tricalcium phosphate (GTG) membrane using carbodiimide. We investigated the effects of these growth factors released from the GTG composites on cultured PC12 cells and sciatic nerve regeneration across a 10-mm-long gap in rats. In PC12 cell culture, the total protein content and MTT assay indicated more cell attachment on the composites modified with growth factors. The IGF-1 group showed a higher survival promotion effect on PC12 cells than did BDNF and NGF groups. On the other hand, NGF released from the composite showed the highest level of neuritogenesis for PC12 cells in neurite outgrowth assay. In the animal study, the GTG conduits modified with various growth factors were well tolerated by the host tissue. In the regenerated nerves, the number of the axons per unit area of the BDNF group was significantly higher than that of NGF and GTG groups but similar to that of IGF-1 group. However, the average axon size was the largest in NGF group. This result was in concordance with the neurite outgrowth assay in which NGF showed the highest neuritogenic potential. In the assessment of motor and sensory recovery after nerve repair, conduits modified with various neurotrophic factors showed a more favorable outcome in compound muscle action potential. The BDNF group had a better gastrocnemic muscle weight ratio than blank GTG repair. Nevertheless, the different effects of GTG conduits modified with various neurotrophic factors on functional recovery cannot be simply illustrated in the sciatic function index. 相似文献
7.
BACKGROUND: Articular cartilage injuries can result from a variety of causes. Conventional therapy cannot obtain the optimal clinical results. Low-intensity pulsed ultrasound has been shown to promote the repair of injured articular cartilage.
OBJECTIVE: To investigate the effects of low-intensity pulsed ultrasound on the repair of injured articular cartilage.
METHODS: Twenty New Zealand white rabbits were used to establish knee arthritis models and equally randomized into study and control groups, respectively. Rabbits in the study group received low-intensity pulsed ultrasound treatment, and sham low-intensity pulsed ultrasound treatment was given in the control group. At 8 weeks after treatment, pathological change and histological scores in articular cartilage tissue collected from both groups were determined. Moreover, the ultrastructure and type II collagen expression of chondrocytes were determined. Matrix metalloproteinase-13 mRNA expression was detected by quantitative real-time PCR.
RESULTS AND CONCLUSION: At 8 weeks after treatment, toluidine blue staining showed a disordered arrangement of cells, decreased number of cartilage cells in each layer and cluster in the control group. Light disordered arrangement of cells, decreased appearance of the superficial layer cells and the cluster phenomenon were observed in the study group. Articular cartilage tissue scores were significantly decreased in the study group compared with the control group (P < 0.05). The chondrocytes were small, enlarged intracellular mitochondria and rough endoplasmic reticulum, cytoplasmic swelling, collagen fibrils coarse, well developed Golgi apparatus, and nuclear fragmentation were observed in the control group. In addition, the normal structure of organelles disappeared and cell degeneration was observed in the control group. In the study group, the size of chondrocytes and the Golgi complex and other organelles were normal, and the protein polysaccharide granules were observed in the cytoplasm and membrane. The mRNA expression of matrix metalloproteinase-13 in the study group was significantly lower than that in the control group (P < 0.05). Type II collagen immunoreactivity in the study group was stronger than that in the control group. No incision infection, suppuration, red swelling appeared in all rabbits. Our results suggest that low-intensity pulsed ultrasound can be used for the treatment of articular cartilage injury by alleviating the degradation of collagen type II and inhibiting the expression of matrix metalloproteinase-13.
中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程 相似文献
8.
背景:软骨是一种无血管的组织,软骨损伤后自身修复能力有限。当前用于治疗关节软骨损伤的方法从保守治疗到手术治疗多种多样。随着组织工程技术的发展,关节软骨的修复又进入了新的高度。
目的:综述组织工程方法修复软骨损伤的新进展。
方法:由第一作者在2013年5月应用计算机检索2000至2013年PubMed 数据库及CNKI 数据库,英文以“cartilage tissue engineering,cartilage defect;stem cell,scaffold;growth factor”为关键词,中文以“软骨组织工程,软骨缺损,干细胞,支架,生长因子”为关键词,选择内容与软骨组织工程、软骨损伤修复相关的文章,同一领域文献则选择近期发表或发表在权威杂志文章,共纳入64篇文献。
结果与结论:软骨组织工程三大要素——种子细胞、支架和细胞因子,三者必须协调发展和互利。现阶段组织工程方法修复关节软骨损伤的研究虽已取得很大进展,但大多停留于实验探索阶段,尚未应用于临床。随着新材料的不断研发,新的组织工程软骨修复材料将兼顾材料学和生物科学的需要,使其更接近机体自身组织生物学特性。在新的技术支持下,动物实验研究也将向临床试验转变,使关节软骨损伤的治疗取得突破性进展。 相似文献
9.
Although several treatments for cartilage repair have been developed and used in clinical practice the last 20 years, little is known about the mechanisms that are involved in the formation of repair tissue after these treatments. Often, these treatments result in the formation of fibrocartilaginous tissue rather than normal articular cartilage. Because the repair tissue is inferior to articular cartilage in terms of mechanical properties and zonal organization of the extracellular matrix, complaints of the patient may return. The biological and functional outcome of these treatments should thus be improved. For this purpose, an in vitro model allowing investigation of the involved repair mechanisms can be of great value. We present the development of such a model. We used bovine osteochondral biopsies and created a system in which cartilage defects of different depths can be studied. First, our biopsy model was characterized extensively: we studied the viability by means of lactate dehydrogenase (LDH) excretion over time and we investigated expression of cartilage-related genes in osteochondral biopsies and compared it with conventional cartilage-only explants. After 28 days of culture, LDH was detected at low levels and mRNA could be retrieved. The expression of cartilage-related genes decreased over time. This was more evident in cartilage-only explants, indicating that the biopsy model provided a more stable environment. We also characterized the subchondral bone: osteoclasts and osteoblasts were active after 28 days of culture, which was indicated by tartrate acid phosphatase staining and alkaline phosphatase measurements, respectively, and matrix deposition during culture was visualized using calcein labeling. Second, the applicability of the model was further studied by testing two distinct settings: (1) implantation of chondrocytes in defects of different depths; (2) two different seeding strategies of chondrocytes. Differences were observed in terms of volume and integration of newly formed tissue in both settings, suggesting that our model can be used to model distinct conditions or even to mimic clinical treatments. After extensive characterization and testing of our model, we present a representative and reproducible in vitro model that can be used to evaluate new cartilage repair treatments and study mechanisms in a controlled and standardized environment. 相似文献
10.
The elastic network of articular cartilage was investigated by immunohistochemistry using specific antibodies to elastin and fibrillin‐1. Articular cartilage was dissected from defined regions of bovine metacarpophalangeal joints. Elastin fibres and microfibrils were dual‐immunostained by labelling with distinct fluorescent dyes. A conventional fluorescence microscope combined with a polarized light filter was used to study the organization and degree of colocalization of elastin fibres, microfibrils and of the collagen network. We observed an elaborately organized elastic network. In the uppermost superficial zone, where few cells were present, elastin fibres and microfibrils formed a dense three dimensional network showing some degree of colocalization. The thickness and organization of this elastic network varied dramatically from region to region and was most extensive in the metacarpal palmar region. In the middle and deep zones, very few elastin fibres were observed but microfibrils formed a network in the inter‐territorial matrix and dense network around the cells. Our finding of a three dimensional network of dense, well organized elastin fibres and microfibrils in the surface zone of the articular cartilage matrix, and a dense network of microfibrils around the cells deeper into the tissue suggests the elastic network could play both a mechanical and a biological role in articular cartilage. 相似文献
11.
It is well-established that cartilage grows by a combination of matrix secretion, cell hypertrophy and cell proliferation. The extent to which this growth is by appositional, as opposed to interstitial mechanisms, however, remains unclear. Using the knee joints of the marsupial Monodelphis domestica to study cartilage growth, we have combined an immunohistochemical study of the TGF-β family of cartilage growth and differentiation factors between 30 days postpartum to 8 months, together with a stereological analysis of cartilage morphology during growth. Furthermore, to gain an insight into the generation of the characteristic zones within cartilage, we have examined the effects of intra-articular administration of bromodeoxyuridine, an agent that is incorporated into DNA during cell division and blocks further cell cycling. During early growth, TGF-β2 and -β3 were widely expressed but TGF-β1 was less so. After the formation of the secondary centre of ossification, all isoforms became more restricted to the upper half of the tissue depth and their distribution was similar to that previously described for IGFs, and PCNA-positive cells. Stereological analysis of tissue sections from the femoral condylar cartilage at 3 and 6 months showed that there was a 17% increase in total cartilage volume but a 31% decrease in cell density on a unit volume basis. Finally, cell-cycle perturbation with BrDU, which was injected into the knee joints of 3-month-old animals and analysed 1 and 4 months post-injection, revealed that the chondrocytes occupying the transitional zone were depleted 1 month post-injection, resulting in thinning of the articular cartilage. This effect was reversed 4 months post-injection. Immunohistochemical analysis revealed that BrDU-treatment altered the expression patterns of all TGF-β isoforms, with a marked reduction in labelling of TGF-β1 and -β3 isoforms in the upper half of the cartilage depth. Overall, the data lends further support to the notion of articular cartilage growing by apposition from the articular surface rather than by interstitial mechanisms. Accepted: 12 February 2001 相似文献
12.
Articular chondrocytes can synthesize new cartilaginous matrix in vivo that forms functional bonds with native cartilage. Other sources of chondrocytes may have a similar ability to form new cartilage with healing capacity. This study evaluates the ability of various chondrocyte sources to produce new cartilaginous matrix in vivo and to form functional bonds with native cartilage. Disks of articular cartilage and articular, auricular, and costal chondrocytes were harvested from swine. Articular, auricular, or costal chondrocytes suspended in fibrin glue (experimental), or fibrin glue alone (control), were placed between disks of articular cartilage, forming trilayer constructs, and implanted subcutaneously into nude mice for 6 and 12 weeks. Specimens were evaluated for neocartilage production and integration into native cartilage with histological and biomechanical analysis. New matrix was formed in all experimental samples, consisting mostly of neocartilage integrating with the cartilage disks. Control samples developed fibrous tissue without evidence of neocartilage. Ultimate tensile strength values for experimental samples were significantly increased (p < 0.05) from 6 to 12 weeks, and at 12 weeks they were significantly greater (p < 0.05) than those of controls. We conclude that articular, auricular, and costal chondrocytes have a similar ability to produce new cartilaginous matrix in vivo that forms mechanically functional bonds with native cartilage. 相似文献
13.
BACKGROUND: It has been reported that tissue-engineered acellular matrix can induce and promote epithelial cells and smooth muscle cells to evolve into a part of body in vivo compared with the normal extracellular matrix.
OBJECTIVE: To investigate the effect of tissue-engineered acellular matrix in articular cartilage repair.
METHODS: Totally 30 New Zealand rabbits were randomly allotted to fibroid tissue and acellular matrix groups (n=15 per group), and then articular cartilage defect models, 4 mm in diameter, were established at the white rabbit femoral condyle. Acellular cartilage matrix scaffold was prepared using bovine knee cartilage, and model rats in the acellular matrix group were repaired with acellular cartilage matrix scaffold and the others in the fibroid tissue group repaired with fibroid tissues. Finally, repair effects between two groups were compared.
RESULTS AND CONCLUSION: The dark blue and porous tissue-engineered acellular matrix material could be found, with a diameter of 5 mm and moderate hardness, and exhibited certain flexibility after cross-linking. Hematoxylin-eosin staining showed that cell debris, blue-stained nuclear materials and residual extracellular matrix disappeared. Toluidine blue staining found that the porosity of the blue scaffold was 90%, and the swelling ratio was (1 314±337)%. The absorbance value in the acellular matrix group was significantly higher than that in the fibroid tissue group at 1, 3, 5, 7 and 9 days ( P < 0.05). In the fibroid tissue group, defects filled with newborn fibrous scars were overt. By contrast, in the acellular matrix group, the white tissues covered the defect region with smooth surface, and the wound was basically healed, with an unclear boundary after 12 weeks. Moreover, blue-stained, small flattened cells appeared, subchondral bone structure was connected well with the cartilage, and the scaffold was directly degraded. In conclusion, the tissue-engineered acellular matrix material exhibits good biocompatibility, cell adsorption and hydrophilicity, and can promote the defect repair after articular cartilage injury. Therefore, it is a suitable substitute for articular cartilage repair. 相似文献
14.
Auricular reconstruction remains a major challenge facing reconstructive surgeons owing to the complexity of autogenous transplants. In this study, the development of a three-dimensional custom-made polyurethane (PU) auricular implant with hydroxyapatite (HA) coating is described. The PU implant was produced by computerized tomography (CT) scanning and indirect rapid prototyping. To improve the physiological response of the implant, the PU prototype was coated with a microrough, homogenous layer of HA by a novel solvent-compression coating method. Bioactivity of the HA coated PU substrates was confirmed by apatite formation on the HA coating after 9 days in revised simulated body fluid (pH 7.4). Adhesion strength of the HA coating to the PU surface using the tensile pull-off test revealed partial failure of the coating with an average tensile strength of 1.6 MPa. As an initial stage indication of cytocompatibility for a soft tissue application, in vitro cell culturing on the HA-coated PU substrates using Graham 293 fibroblast cells was performed. After 24 and 72 h, the HA coated surfaces displayed significantly higher cell numbers and metabolically active cells compared with the virgin uncoated PU surfaces. This indicates that HA coated PU surfaces are cytocompatible towards fibroblasts and could potentially be applied to auricular cartilage tissue replacement. 相似文献
15.
背景:将羟基磷灰石与聚乙烯醇水凝胶复合之后应用于软骨缺损修复中,可在软骨连接部位产生良好的生物活性,有效促进骨细胞的生长,提高植入材料的稳定性和生物活性。
目的:观察聚乙烯醇/羟基磷灰石复合水凝胶修复兔膝关节软骨缺损的组织相容性。
方法:取20只新西兰大白兔,随机分为空白对照组( n=6)与实验组( n=14),构建单侧膝关节软骨缺损模型,空白对照组不予以修复,实验组予以聚乙烯醇/羟基磷灰石复合水凝胶修复。术后4,8,12周获取膝关节标本进行大体观察及组织学观察。
结果与结论:空白对照组关节软骨面在术后12周内始终未得到修复,软骨下缺损存在肉芽组织充填现象,组织学也未见明显修复。实验组术后4周可见聚乙烯醇/羟基磷灰石复合水凝胶填充于缺损处,与周围软骨组织之间连接紧密,且存在清晰的界限,未出现细胞长入情况;至12周时,聚乙烯醇/羟基磷灰石复合水凝胶呈白色、半透明状,表面平坦,与周围软骨组织之间存在清晰界限,两者交界面存在软骨细胞大量增殖现象,与周围组织发生紧密结合,二者之间无间隙,底部与软骨下骨紧密连接,并存在骨样组织长入。表明聚乙烯醇羟基磷灰石复合水凝胶修复兔膝关节软骨缺损具有良好的组织相容性。
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 相似文献
16.
The aim of this study was to evaluate the potential for restoration of a large cartilage defect in the goat knee with hydroxyapatite (HA) loaded with chondrocytes. Isolated chondrocytes were suspended in fibrin glue, seeded on top of the HA, and then the composite graft was implanted in the defect. After transplantation, cell behaviour, newly synthesised matrix and the HA–glue interface were assessed histologically after 2, 4, 12, 26 and 52 weeks. Special attention was paid to the incorporation process of HA in the subchondral bone and interactions between this biomaterial and the fibrin-glue–chondrocyte suspension. Chondrocytes in the glue proved to survive the transplantation procedure and produced new metachromatically stained matrix two weeks after implantation. The glue–cell suspension had penetrated the superficial porous structure of the HA. Four weeks after surgery, islands of hyaline-like cartilage were observed at the HA–glue interface. A layer of fibrous tissue was formed surrounding the HA graft, resulting in a relatively instable fixation of the HA in the defect. This instability of the graft in the defect, possibly together with early weight bearing, resulted in a gradual loss of the newly formed hyaline cartilage-like repair tissue. Progressive resorption of the HA occurred without any sign of active bone remodelling from the host site. One year after surgery part of the defect which extended down to the cancellous bone had been predominantly restored with newly formed lamellar bone. Only small HA remnants were still present at the bottom of the original defect. Resurfacing of the joint had occurred with fibrocartilaginous repair tissue. The absence of adequate fixation capacity of the HA near the joint space resulted in a relative instability of the graft with progressive resorption. Therefore, HA is not a suitable biomaterial to facilitate the repair of large articular cartilage defects. 相似文献
17.
Recent studies have shown that adult human articular cartilage contains stem-like cells within the native structure. In this study, we aimed to determine the localization of putative stem cell markers such as CD90, STRO-1, OCT-3/4, CD105 and CD166 in adult human articular cartilage tissue sections and demonstrate the expression of these markers within the expanded surface zone colony-forming (CF) cells and evaluate their differentiation potential. Biopsy samples were either fixed immediately for immunohistochemical analyses or processed for in vitro cell culture. Immunohistochemical and flow cytometry analyses were performed by using CD90, STRO-1, OCT-3/4, CD105 and CD166 antibodies. Isolated colony-forming (CF) cells were further stimulated, by using the appropriate growth factors in their pellet culture, to obtain cartilage, bone and adipose lineages. We observed that the expression of the stem cell markers were in various zones of the human adult cartilage. Flow cytometry results showed that in CF cells the expression of CD90 and CD166 was high, while OCT-3/4 was low. We also determined that CF cells could be stimulated towards cartilage, bone and adipose lineages. The results of this research support the idea that the resident stem-like cells in adult human articular cartilage express these putative stem cell markers, but further experimental investigations are needed to determine the precise localization of these cells. 相似文献
18.
背景:不同生物材料制备的复合软骨支架其修复软骨缺损也各具特点。
目的:探讨不同生物材料制备复合支架的组织工程学特性及其修复关节软骨缺损的性能评价。
方法:以“软骨组织工程,生物材料,工程软骨,复合支架”为中文关键词,以“tissue enginneering,articular cartilage,scaffold material”为英文关键词,采用计算机检索中国期刊全文数据库、PubMed数据库(1993-01/2010-11)相关文章。纳入复合支架材料-细胞复合物修复关节软骨损伤等相关的文章,排除重复研究或Meta分析类文章。
结果与结论:复合支架是当前软骨组织工程中应用较多的支架,它是将具有互补特征的生物相容性可降解支架,按一定比例和方式组合,设计出结构与性能优化的复合支架。较单一支架材料具有更好的生物相容性和一定强度的韧性,较好的孔隙和机械强度。复合支架的制备不仅包括同一类生物材料的复合,还包括不同类别生物材料之间的交叉复合。可分为纯天然支架材料、纯人工支架材料以及天然与人工支架材料的复合等3类。复合支架使生物材料具有互补特性,一定程度上满足了理想生物材料支架应具的综合特点,但目前很多研究仍处于实验阶段,还有一些问题有待于解决,如不同材料的复合比例、复合工艺等。 相似文献
19.
目的:总结组织工程技术及生物材料在防治运动性关节软骨损伤中的应用特点。
方法:以“关节软骨,组织工程技术,生物材料”为中文关键词,以“tissue enginneering, articular cartilage, scaffold material”为英文关键词,采用计算机检索Pubmed数据库(http://www.ncbi.nlm.nih.gov/PubMed)及维普数据库(http://www.cqvip.com/)1993-01/2010-10的相关文章,排除重复研究或Meta分析类文章。以23篇文献为主,重点对修复运动性关节软骨损伤种子细胞、支架材料、细胞因子及其性能进行讨论。
结果:计算机初检得到104篇文献,根据纳入排除标准,对组织工程软骨的种子细胞、生物支架材料以及用于组织工程中的细胞因子进行总结与分析。种子细胞是制约组织工程软骨进一步临床应用的首要因素,目前常采用的种子细胞有软骨细胞、骨髓基质干细胞和胚胎干细胞等;生物支架材料包括天然材料和人工合成可降解聚合物等;用于软骨组织工程的生长因子主要包括转化生长因子、骨形成蛋白、成纤维细胞生长因子、胰岛素样生长因子等。
结论:迄今为止,无论是工程软骨的种子细胞、支架材料、培养环境等还没有任何一种材料被认为最理想,寻求一种具有良好性能的组织工程化关节软骨是未来研究的重点。但目前很多研究仍处于实验阶段,还有一些问题有待于解决,特别是组织工程细胞支架材料植入体内后,材料的降解与细胞功能发挥是否同步,会不会产生遗传物质改变、基因表达或基因突变等问题,将其应用于临床更需要相关学者专家不断的实践和探索。 相似文献
20.
PURPOSE: To determine the effect of polyglycolic acid (PGA)-polylactic acid (PLA) co-polymer pads with calcium alginate on chondrogenic gene expression for chondrocytes cultured in vitro. We also evaluated the ability of these absorbable pads with alginate to deliver chondrocytes and influence osteochondral defect repair in vivo in immature rabbit knees. METHODS: Rabbit rib chondrocytes were suspended in calcium alginate and co-polymer pads composed of either 47.5/52.5 PGA-PLA or 90/10 PGA-PLA at two different cell concentrations and cultured in vitro for 1, 3, and 5 days. Analysis was performed using RT-PCR for chondrogenic gene expression of aggrecan, type II collagen, and type I collagen. Cells labeled with a traceable green fluorescent protein (GFP) marker in vitro were suspended within the pads to analyze for dispersion and attachment to the pad. An in vivo study was performed in which full-thickness (3x4mm(2)) osteochondral defects were made in 60 rabbit knees. The study comprised four treatment groups based on the type of implant into the defect (empty, alginate alone, or either type of co-polymer pad) and harvested at either 6 or 12 weeks. Two independent blinded observers analyzed and scored the defects grossly and histologically. RESULTS: In vitro analysis of the chondrocytes after 1, 3, and 5 days in culture showed no statistical differences between the types of PGA/PLA co-polymer pad with regard to expression of aggrecan, type II collagen, or type I collagen. However, although statistically insignificant, the expression of aggrecan and type II collagen was found to be greater than that for type I collagen in both types of pads, confirming the chondrogenic effect of suspension culture for this system. The addition of alginate to polymer pads allowed costal chondrocytes to be implanted in vivo, as evidenced by the attachment of the cells to the fibers and the uniform dispersion of the GFP-labeled cells through the pad as seen on fluorescent microscopy. Histologic results showed improved scores for the 47.5/52.5 PGA-PLA group (21.3) and the 90/10 PGA-PLA group (18.3) when compared to empty (15.3) or alginate alone (15.1) defects at 12 weeks. CONCLUSION: The addition of calcium alginate to the co-polymer pads offers a new approach to deliver cells to an osteochondral defect and may enhance cartilage regeneration. Future application of this model may allow for an arthroscopic delivery system to assist the healing of cartilage defects. 相似文献
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