Influence of nanofibers on growth and gene expression of human tendon derived fibroblast

Abstract

BackgroundRotator cuff tears are a common and frequent lesion especially in older patients. The mechanisms of tendon repair are not fully understood. Common therapy options for tendon repair include mini-open or arthroscopic surgery. The use of growth factors in experimental studies is mentioned in the literature. Nanofiber scaffolds, which provide several criteria for the healing process, might be a suitable therapy option for operative treatment. The aim of this study was to explore the effects of nanofiber scaffolds on human tendon derived fibroblasts (TDF's), as well as the gene expression and matrix deposition of these fibroblasts.MethodsNanofibers composed of PLLA and PLLA/Col-I were seeded with human tendon derived fibroblasts and cultivated over a period of 22 days under growth-inductive conditions, and analyzed during the course of culture, with respect to gene expression of different extra cellular matrix components such as collagens, bigylcan and decorin. Furthermore, we measured cell densities and proliferation by using fluorescene microscopy.ResultsPLLA nanofibers possessed a growth inhibitory effect on TDF's. Furthermore, no meaningful influence on the gene expression of collagen I, collagen III and decorin could be observed, while the expression of collagen X increased during the course of cultivation. On the other hand, PLLA/Col-I blend nanofibers had no negative influence on the growth of TDF's. Furthermore, blending PLLA nanofibers with collagen had a positive effect on the gene expression of collagen I, III, X and decorin. Here, gene expression indicated that focal adherence kinases might be involved.ConclusionThis study indicates that the use of nanofibers influence expression of genes associated with the extra cellular matrix formation. The composition of the nanofibers plays a critical role. While PLLA/Col-I blend nanofibers enhance the collagen I and III formation, their expression on PLLA nanofibers was more comparable to controls. However, irrespective of the chemical composition of the fibres, the collagen deposition was altered, an effect which might be associated with a decreased expression of biglycanes.

The past, present and future in scaffold-based tendon treatments

“…Using a wet extrusion system, micro-scale collagen fibres (Figure 3) were first produced in late 1970's and since then numerous papers have demonstrated that this process gives rise to fibres with ultrastructure characteristics, physical and mechanical properties similar to native tendon [257,259,[357][358][359]. Further in vitro analysis demonstrated that such materials, largely attributed to their surface features, not only facilitate bidirectional cell alignment, but also maintain tenogenic phenotype in vitro [360]. Further, preclinical experimentation in peripheral nerve repair and in tendon small and large animal models enhanced the clinical potential of these fibres, as judged by improved structural alignment and biomechanics [361][362][363].…”

“…Despite the huge progress that has been achieved to-date, synthetic materials are yet to meet the requirements for functional tendon tissue remodelling [426]. The use of PLGA and PCL has been problematic, with in vitro data showing poor tendon cell adhesion, reduced proliferation rate and phenotypic drift [360,427]; with preclinical in vivo data showing poor cellular infiltration and mechanical properties compared to controls [428]; and with clinical evaluations showing problems with cell lysis, anchor failure, macrophage stimulation and allergic responses [429,430]. This high failure rate has been attributed to their hydrophobic nature and their lack of cell recognition signals that prevents cell attachment [391,[431][432][433][434].…”

A novel Bruch's membrane-mimetic electrospun substrate scaffold for human retinal pigment epithelium cells

“…After EMT, RPE cells lose many of the RPE-specific characteristics, including expression of key genes. Additionally, according to Theisen's results, gene expression in human cells can also be altered by nanofibers [59]. We tested for the expression of 9 marker genes, and ARPE-19 cells cultured on the different membranes showed equivalent levels of expression to those cultured on TCP, suggesting that our electrospun membranes did not disturb normal RPE-cell gene expression.…”

A Nanofibrillar Surface Promotes Superior Growth Characteristics in Cultured Human Retinal Pigment Epithelium

“…Reports on proliferation rates altered by nanofiber topographies vary; both increased [11] as well as decreased growth kinetics have been reported [19] . Our study, however, clearly demonstrates faster RPE proliferation on nanofiber surfaces.…”