Gögele Clemens, Hahn Judith, Elschner Cindy, Breier Annette, Schröpfer Michaela, Prade Ina, Meyer Michael, Schulze-Tanzil Gundula
Institute of Anatomy and Cell Biology, Paracelsus Medical University, Nuremberg and Salzburg, Prof. Ernst Nathan Str. 1, 90419 Nuremberg, Germany.
Department of Biosciences, Paris Lodron University of Salzburg, Hellbrunnerstr. 34, 5020 Salzburg, Austria.
Int J Mol Sci. 2020 Feb 8;21(3):1132. doi: 10.3390/ijms21031132.
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---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.
前交叉韧带(ACL)断裂的重建受到自体移植物可用性和供体部位发病率的限制。因此,组织工程化移植物可能在未来成为一种替代方案。本研究旨在确定兔(L)ACL来源的成纤维细胞在绣有聚(L-丙交酯-己内酯)(P(LA-CL))和聚乳酸(PLA)支架上的性能,以探讨ACL重建的组织工程方法。测试了通过气相氟化对P(LA-CL)/PLA进行表面改性以及使用1-乙基-3-(3-二甲基氨基丙基)碳二亚胺(EDC)或六亚甲基二异氰酸酯(HMDI)对胶原泡沫进行交联对细胞粘附、生长和基因表达的影响。实验使用了动态或静态接种LACL来源成纤维细胞的绣有P(LA-CL)/PLA的支架。评估了支架的细胞相容性、细胞存活率、数量、代谢活性、超微结构和硫酸化糖胺聚糖(sGAG)合成。定量实时聚合酶链反应(QPCR)显示了I型胶原(COL1A1)、核心蛋白聚糖(DCN)、腱生蛋白C(TNC)、莫霍克蛋白(MKX)和腱调节蛋白(TNMD)的基因表达。所有测试的支架都具有高度的细胞相容性。与用EDC交联或未进行任何功能化处理的支架相比,在HMDI交联和氟化的支架中检测到细胞数量显著更高,且细胞定植的支架表面积更大。相比之下,对照组中的sGAG合成更高。尽管未达到显著水平,但在含有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总体上更高。LACL来源的成纤维细胞在补充有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。