Chondrogenic phenotype in responses to poly(ɛ-caprolactone) scaffolds catalyzed by bioenzymes: effects of surface topography and chemistry

Abstract

Biodegradable poly(ε-caprolactone) (PCL) has been increasingly investigated as a promising scaffolding material for articular cartilage tissue repair. However, its use can be limited due to its surface hydrophobicity and topography. In this study, 3D porous PCL scaffolds fabricated by a fused deposition modeling (FDM) machine were enzymatically hydrolyzed using two different biocatalysts, namely Novozyme®435 and Amano lipase PS, at varied treatment conditions in a pH 8.0 phosphate buffer solution. The improved surface topography and chemistry of the PCL scaffolds were anticipated to ultimately boost the growth of porcine articular chondrocytes and promote the chondrogenic phenotype during cell culture. Alterations in surface roughness, wettability, and chemistry of the PCL scaffolds after enzymatic treatment were thoroughly investigated using several techniques, e.g., SEM, AFM, contact angle and surface energy measurement, and XPS. With increasing enzyme content, incubation time, and incubation temperature, the surfaces of the PCL scaffolds became rougher and more hydrophilic. In addition, Novozyme®435 was found to have a higher enzyme activity than Amano lipase PS when both were used in the same enzymatic treatment condition. Interestingly, the enzymatic degradation process rarely induced the deterioration of compressive strength of the bulk porous PCL material and slightly reduced the molecular weight of the material at the filament surface. After 28 days of culture, both porous PCL scaffolds catalyzed by Novozyme®435 and Amano lipase PS could facilitate the chondrocytes to not only proliferate properly, but also function more effectively, compared with the non-modified porous PCL scaffold. Furthermore, the enzymatic treatments with 50 mg of Novozyme®435 at 25 °C from 10 min to 60 min were evidently proven to provide the optimally enhanced surface roughness and hydrophilicity most significantly favorable for induction of chondrogenic phenotype, indicated by the greatest expression level of cartilage-specific gene and the largest production of total glycosaminoglycans.