Chang Nai-Jen, Jhung Yi-Ru, Issariyakul Nontapot, Yao Chih-Kai, Yeh Ming-Long
a Institute of Biomedical Engineering, National Cheng Kung University , 1 University Road , Tainan City , 701 , Taiwan.
J Biomater Sci Polym Ed. 2012;23(17):2133-51. doi: 10.1163/092050611X611648. Epub 2012 May 8.
Scaffold surface properties and mechanical stimuli have been known to have a critical influence on cell proliferation and extracellular matrix synthesis in cultured cells for tissue engineering. Hydrophilic surface and hydrodynamic pressure (HP) stimulation have been shown to have a beneficial effect on chondrocyte activity. The aim of this study was, thus, to assess the synergic influences of HP and hydrophilic coating on cell activity using primary porcine chondrocytes inoculated in hydrophilic-coated poly(lactide-co-glycolide) (PLGA) sponge scaffolds. The natural materials hyaluronic acid (HA), chitosan and HA/chitosan were cross-linked on porous PLGA as a hydrophilic surface modification. HP was applied to scaffolds at an amplitude of 2.24 MPa and a frequency of 0.1 Hz for 30 min per day, twice a week, over a period of 28 days. Cell activities were determined by the MTS assay and the dimethylmethylene blue assay for glycosaminoglycan (GAG) quantification. Our results displayed that PLGA coated with both HA and chitosan had the best hydrophilicity (contact angle 49.46°) and initial compressive modulus (1.10 ± 0.13 MPa) among the tested scaffold groups. Additionally, HP stimulation enhanced cell proliferation as well as GAG production (up to 3-fold in culture medium and 15-fold in scaffolds at 28 days compared to static culture of PLGA alone in the scaffold group) in the hydrophilic-coated scaffold groups. The synergistic benefit from hydrophilic coating and HP stimulation may be imperative in regenerating engineered cartilage in the long-term.
已知支架表面特性和机械刺激对组织工程中培养细胞的细胞增殖和细胞外基质合成具有关键影响。亲水性表面和流体动力压力(HP)刺激已被证明对软骨细胞活性有有益影响。因此,本研究的目的是使用接种在亲水性涂层聚(丙交酯-共-乙交酯)(PLGA)海绵支架中的原代猪软骨细胞,评估HP和亲水性涂层对细胞活性的协同影响。天然材料透明质酸(HA)、壳聚糖和HA/壳聚糖在多孔PLGA上交联作为亲水性表面改性。以2.24 MPa的振幅和0.1 Hz的频率对支架施加HP,每天30分钟,每周两次,持续28天。通过MTS测定法和用于糖胺聚糖(GAG)定量的二甲基亚甲基蓝测定法测定细胞活性。我们的结果显示,在测试的支架组中,同时涂覆有HA和壳聚糖的PLGA具有最佳的亲水性(接触角49.46°)和初始压缩模量(1.10±0.13 MPa)。此外,在亲水性涂层支架组中,HP刺激增强了细胞增殖以及GAG的产生(与支架组中单独PLGA的静态培养相比,在28天时培养基中高达3倍,支架中高达15倍)。亲水性涂层和HP刺激的协同益处可能对长期再生工程软骨至关重要。