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Bone Tissue Engineering with Adipose-Derived Stem Cells in Bioactive Composites of Laser-Sintered Porous Polycaprolactone Scaffolds and Platelet-Rich Plasma

机译:脂肪烧结干多孔聚己内酯支架和富含血小板血浆的生物活性复合材料中脂肪组织干细胞的骨组织工程。

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摘要

Three-dimensional porous polycaprolactone (PCL) scaffolds with consistent inter-pore channels, 83% porosity and 300–400 μm pore size were fabricated via selective laser sintering. The PCL scaffold was combined with platelet-rich plasma (PRP) to form a bioactive composite and studied for potential application in bone tissue engineering using porcine adipose-derived stem cells (PASCs). The PCL/PRP/PASCs construct showed enhanced cell seeding efficiency and synergistically increased the differentiation capability of PASCs in osteogenic medium toward the osteoblast lineage, judging from elevated alkaline phosphatase activity and up-regulated osteogenic genes expression. For in vivo study, a 3 cm × 3 cm mandible defect was created in pigs and reconstructed by implanting acellular PCL scaffolds or PCL/PRP/PASCs constructs. Both groups showed new bone formation, however, the new bone volume was 5.1 times higher for PCL/PRP/PASCs 6 months post-operation. The bone density was less and loose in the acellular PCL group and the Young’s modulus was only 29% of normal bone. In contrast, continued and compact bone formation was found in PCL/PRP/PASCs and the Young’s modulus was 81% that of normal bone. Masson’s trichrome stain, immunohistochemical analysis of osteocalcin and collagen type I also confirmed new bone formation.
机译:通过选择性激光烧结制备了具有一致的孔间通道,83%孔隙率和300-400μm孔径的三维多孔聚己内酯(PCL)支架。将PCL支架与富血小板血浆(PRP)结合以形成具有生物活性的复合物,并研究了使用猪脂肪干细胞(PASC)在骨组织工程中的潜在应用。从碱性磷酸酶活性升高和成骨基因表达上调来看,PCL / PRP / PASCs构建体显示出更高的细胞接种效率,并协同增强了成骨培养基中PASCs向成骨细胞谱系的分化能力。为了进行体内研究,在猪中创建了一个3 cm×3 cm的下颌骨缺损,并通过植入脱细胞PCL支架或PCL / PRP / PASCs构建体进行了重建。两组均显示新的骨形成,但是,术后6个月PCL / PRP / PASC的新骨量高出5.1倍。无细胞PCL组的骨密度较小且疏松,杨氏模量仅为正常骨的29%。相反,在PCL / PRP / PASC中发现了连续而致密的骨形成,杨氏模量是正常骨的81%。 Masson的三色染色剂,骨钙素和I型胶原的免疫组织化学分析也证实了新的骨形成。

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