Engineered substrates incapable of induction of chondrogenic differentiation compared to the chondrocyte imprinted substrates

Abstract

It is well established that surface topography can affect cell functions. However, finding a reproducible and reliable method for regulating stem cell behavior is still under investigation. It has been shown that cell imprinted substrates contain micro- and nanoscale structures of the cell membrane that serve as hierarchical substrates, can successfully alter stem cell fate. This study investigated the effect of the overall cell shape by fabricating silicon wafers containing pit structure in the average size of spherical-like chondrocytes using photolithography technique. We also used chondrocyte cell line (C28/I2) with spindle-like shape to produce cell imprinted substrates. The effect of all substrates on the differentiation of adipose-derived mesenchymal stem cells (ADSCs) has been studied. The AFM and SEM images of the prepared substrates demonstrated that the desired shapes were successfully transferred to the substrates. Differentiation of ADSCs was investigated by immunostaining for mature chondrocyte marker, collagen II, and gene expression of collagen II, Sox9, and aggrecan markers. C28/I2 imprinted substrate could effectively enhanced chondrogenic differentiation compared to regular pit patterns on the wafer. It can be concluded that cell imprinted substrates can induce differentiation signals better than engineered lithographic substrates. The nanostructures on the cell-imprinted patterns play a crucial role in harnessing cell fate. Therefore, the patterns must include the nano-topographies to have reliable and reproducible engineered substrates.

It Takes Two to Tango: Controlling Human Mesenchymal Stromal Cell Response via Substrate Stiffness and Surface Topography

“…The silicone waffle pattern did not enhance chondrogenic differentiation of AD-hMSC. In comparison, the patterned PDMS imitating the shape of a chondrocyte promoted the expression of chondrogenic markers including collagen II, Sox9, and aggrecan [260] Chondroitin sulfate coated PCL film, nanopillar, 250 nm diameter and 500 nm pitch; nanoholes, 225 nm diameter and 400 nm pitch; nanogrill, 250 nm line and 250 nm space (nanoimprinting), BM-hMSC…”

Improving Efficiency of Direct Pro-Neural Reprogramming: Much-Needed Aid for Neuroregeneration in Spinal Cord Injury

“…Briefly, the authors used polydimethylsiloxane silicone substrates to capture and recapitulate the topology of the target human NPCs and, subsequently, cultured adipose-derived stem cells on these substrates, which led to changes in the cell morphology and the upregulation of several markers of neural SCs as well as early neuronal markers [118]. Later, another work confirmed this observation: that cell-imprinted patterns may harness SCs toward a particular cell fate [119]. The sustained delivery of neurotrophic factors (brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3), nerve growth factor (NGF), and others) is another approach to SCI management (as reviewed in [120]).…”