Enhanced Growth of Lapine Anterior Cruciate Ligament-Derived Fibroblasts on Scaffolds Embroidered from Poly(L-lactide-co-ε-caprolactone) and Polylactic Acid Threads Functionalized by Fluorination and Hexamethylene Diisocyanate Cross-Linked Collagen Foams
Enhanced Growth of Lapine Anterior Cruciate Ligament-Derived Fibroblasts on Scaffolds Embroidered from Poly(L-lactide-co-ε-caprolactone) and Polylactic Acid Threads Functionalized by Fluorination and Hexamethylene Diisocyanate Cross-Linked Collagen Foams
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DOI：
10.3390/ijms21031132
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发表时间：
2020-02-01
影响因子：
5.6
通讯作者：
Schulze-Tanzil, Gundula
中科院分区：
文献类型：
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作者：
Goegele, Clemens;Hahn, Judith;Schulze-Tanzil, Gundula
断裂前交叉韧带（ACLs）的重建受到自体移植物的可用性和供体部位发病率的限制。因此，组织工程移植在未来可能是一种替代方法。本研究旨在确定lapine (L) ACL衍生成纤维细胞在绣花聚（L -乳酸-co-epsilon-己内酯）（P(LA-CL)）和聚乳酸（PLA）支架上的性能，并采用组织工程方法重建ACL。通过气相氟化和用乙基碳二亚胺（EDC）或六亚甲基二异氰酸酯（HMDI）交联胶原泡沫对P(LA-CL)/PLA的表面修饰，测试了它们对细胞粘附、生长和基因表达的影响。实验采用刺绣P(LA-CL)/PLA支架进行，该支架动态或静态地植入lacl来源的成纤维细胞。观察支架细胞相容性、细胞存活率、细胞数量、代谢活性、超微结构和硫酸糖胺聚糖（sGAG）合成。定量实时聚合酶链反应（QPCR）显示I型胶原（COL1A1）、decorin （DCN）、tenascin C （TNC）、Mohawk （MKX）和tenomodulin （TNMD）基因的表达。所有测试的支架都具有高度的细胞相容性。与与EDC交联或没有任何功能化的支架相比，在HMDI交联和氟化支架中检测到更高的细胞密度和更大的支架表面积。相比之下，对照组的sGAG合成较高。虽然没有达到显著性水平，但在交联胶原泡沫的氟化支架中，韧带细胞外基质成分和分化标志物的基因表达普遍较高。lacl来源的成纤维细胞在添加了HMDI交联胶原泡沫的氟化支架上保持其分化表型，使其成为ACL组织工程的一个有前途的工具。
Reconstruction of ruptured anterior cruciate ligaments (ACLs) is limited by the availability and donor site morbidity of autografts. Hence, a tissue engineered graft could present an alternative in the future. This study was undertaken to determine the performance of lapine (L) ACL-derived fibroblasts on embroidered poly( L-lactide-co-epsilon-caprolactone) (P(LA-CL)) and polylactic acid (PLA) scaffolds in regard to a tissue engineering approach for ACL reconstruction. Surface modifications of P(LA-CL)/PLA by gas-phase fluorination and cross-linking of a collagen foam using either ethylcarbodiimide (EDC) or hexamethylene diisocyanate (HMDI) were tested regarding their influence on cell adhesion, growth and gene expression. The experiments were performed using embroidered P(LA-CL)/PLA scaffolds that were seeded dynamically or statically with LACL-derived fibroblasts. Scaffold cytocompatibility, cell survival, numbers, metabolic activity, ultrastructure and sulfated glycosaminoglycan (sGAG) synthesis were evaluated. Quantitative real-time polymerase chain reaction (QPCR) revealed gene expression of collagen type I (COL1A1), decorin (DCN), tenascin C (TNC), Mohawk (MKX) and tenomodulin (TNMD). All tested scaffolds were highly cytocompatible. A significantly higher cellularity and larger scaffold surface areas colonized by cells were detected in HMDI cross-linked and fluorinated scaffolds compared to those cross-linked with EDC or without any functionalization. By contrast, sGAG synthesis was higher in controls. Despite the fact that the significance level was not reached, gene expressions of ligament extracellular matrix components and differentiation markers were generally higher in fluorinated scaffolds with cross-linked collagen foams. LACL-derived fibroblasts maintained their differentiated phenotype on fluorinated scaffolds supplemented with a HMDI cross-linked collagen foam, making them a promising tool for ACL tissue engineering.