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组织工程化壳聚糖-明胶复合支架负载人血小板凝胶促进大鼠临界尺寸桡骨骨缺损的再生。

Effectiveness of tissue engineered chitosan-gelatin composite scaffold loaded with human platelet gel in regeneration of critical sized radial bone defect in rat.

机构信息

Department of Pathology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran.

Department of Pathology, School of Veterinary Medicine, Shiraz University, Shiraz, Iran.

出版信息

J Control Release. 2017 May 28;254:65-74. doi: 10.1016/j.jconrel.2017.03.040. Epub 2017 Mar 29.

Abstract

Although many strategies have been utilized to accelerate bone regeneration, an appropriate treatment strategy to regenerate a new bone with optimum morphology and mechanical properties has not been invented as yet. This study investigated the healing potential of a composite scaffold consisting of chitosan (CS), gelatin (Gel) and platelet gel (PG), named CS-Gel-PG, on a bilateral critical sized radial bone defect in rat. Eighty radial bone defects were bilaterally created in 40 Sprague-Dawley rats and were randomly divided into eight groups including untreated, autograft, CS, Gel, CS-PG, Gel-PG, CS-Gel, and CS-Gel-PG treated defects. The bone defects were evaluated clinically and radiologically during the study and their bone samples were assessed by gross and histopathology, histomorphometry, CT-scan, scanning electron microscopy, and biomechanical testing after 8weeks of bone injury. The autograft and CS-Gel-PG groups showed significantly higher new bone formation, density of osseous and cartilaginous tissues, bone volume, and mechanical performance than the defect, CS and Gel-PG groups (P˂0.05). In addition, bone volume, density of osseous and cartilaginous tissues, and numbers of osteons in the CS-Gel-PG group were significantly superior to the CS-PG, CS-Gel and Gel groups (P˂0.05). Increased mRNA levels of alkaline phosphatase, runt-related transcription factor 2, osteocalcin, collagen type 1 and CD31, vascular endothelial growth factor as osteogenic and angiogenic differentiation markers were found with the CS-Gel-PG scaffold by quantitative real-time PCR in vitro after 30days of culturing on bone marrow-derived mesenchymal stem cells. In conclusion, the healing potential of CS-Gel scaffold embedded with PG was comparable to autografting and therefore, it can be offered as an appropriate scaffold in bone tissue engineering and regenerative applications.

摘要

虽然已经有许多策略被用于加速骨再生,但目前还没有发明出一种能够以最佳形态和机械性能再生新骨的适当治疗策略。本研究调查了壳聚糖 (CS)、明胶 (Gel) 和血小板凝胶 (PG) 组成的复合支架 CS-Gel-PG 在大鼠双侧临界尺寸桡骨缺损中的愈合潜力。在 40 只 Sprague-Dawley 大鼠的双侧共创建了 80 个桡骨骨缺损,并将其随机分为 8 组,包括未处理、自体移植物、CS、Gel、CS-PG、Gel-PG、CS-Gel 和 CS-Gel-PG 处理的缺损。在研究过程中,通过临床和放射学评估骨缺损,并在骨损伤 8 周后通过大体和组织病理学、组织形态计量学、CT 扫描、扫描电子显微镜和生物力学测试评估其骨样本。与缺损、CS 和 Gel-PG 组相比,自体移植物和 CS-Gel-PG 组的新骨形成、骨和软骨组织密度、骨体积和机械性能明显更高(P˂0.05)。此外,CS-Gel-PG 组的骨体积、骨和软骨组织密度以及骨单位数量均明显优于 CS-PG、CS-Gel 和 Gel 组(P˂0.05)。通过定量实时 PCR 在体外培养骨髓间充质干细胞 30 天后,在 CS-Gel-PG 支架上发现碱性磷酸酶、Runt 相关转录因子 2、骨钙素、胶原 I 型和 CD31、血管内皮生长因子等成骨和成血管分化标志物的 mRNA 水平升高。总之,PG 嵌入式 CS-Gel 支架的愈合潜力可与自体移植物相媲美,因此可作为骨组织工程和再生应用中的合适支架。

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