Inverted Colloidal Crystal Research Articles

Dual-sized inverted colloidal crystal scaffolds grafted with GDF-8 and Wnt3a for enhancing differentiation of iPS cells toward islet β-cells

BackgroundScaffolds containing two kinds of regular pore size that support differentiation and proliferation of induced pluripotent stem cells (iPSC) are important approaches to artificial pancreas. MethodsThis study aimed to differentiate iPSC in dual-sized gelatin (Gel)-alginate (Alg)-hyaluronic acid (HA) scaffolds containing inverted colloidal crystal (ICC) topography with 70-µm and 160-µm pore sizes for insulin production. The pore surface of dual-sized Gel-Alg-HA ICC scaffolds was grafted with growth differentiation factor 8 (GDF-8) and Wnt3a, and treated with retinoic acid (RA) and noggin to improve the differentiation of iPSC toward endodermic cells, and then islet cells. FindingsAn increasing weight percentage of Gel increased the adhesion efficiency of iPSC. Moreover, Alg and HA helped to promote the proliferation and survival rate of iPSC, which resulted in a higher availability of iPSC for differentiation into insulin-production cells. Immunofluorescence staining, flow cytometry and western blot evidenced that dual-sized Gel-Alg-HA ICC scaffolds grafted with GDF-8 and Wnt3a enhanced the differentiation of iPSC into endodermic cells. When subsequent treatment with RA and noggin, endodermic cells differentiation into insulin-production cells in the scaffolds were improved. Precisely controlling the physical and biomedical properties of dual-sized Gel-Alg-HA ICC scaffolds can guide iPSC to differentiate first into endodermic cells, and then into pancreatic β-cells for diabetic treatment.

IPSCs-laden GDF8-grafted aldehyde hyaluronic acid-polyacrylamide inverted colloidal crystal constructs with controlled release of CHIR99021 and retinoic acid to generate insulin-producing cells

Recent advances in attempting maturation of pancreatic β cells derived from induced pluripotent stem cells (iPSCs) have initiated a gateway to developing innovated materials and protocols for artificial pancreas implantation in diabetic management. In this study, we proposed a tissue-engineering scheme using iPSCs-laden aldehyde hyaluronic acid (AHA)-polyacrylamide (PAAM) inverted colloidal crystal constructs (ICCCs) associated with controlled release of induction molecules to generate insulin-producing cells (IPCs). Based on physicochemical properties, AHA-PAAM inverted colloidal crystal scaffolds (ICCSs) at a 1:1 weight ratio of AHA to PAAM were used, modified with growth and differentiation factor 8, and grafted with CHIR99021-loaded solid lipid nanoparticles (CHIR99021-SLNs-GDF8-AHA-PAAM ICCSs) to differentiate iPSCs into definitive endoderm (DE) cells. Induction with fibroblast growth factor 7-grafted retinoic acid-loaded SLNs (FGF7-RA-SLNs) further differentiated DE cells into insulin-producing cells (IPCs) in the constructs. An introduction of sustained delivery facilitated bioavailability of CHIR99021 and RA during DE and islet formation, and yielded a high quantity of secreted insulin from IPCs after stimulation of glucose. Loaded CHIR99021 and grafted FGF7 acted as supporters to promote the main function of grafted GDF8 and loaded RA in DE and islet growth. From the highest expression of pancreatic and duodenal homeobox 1, we have reached the appropriate concentration of GDF8, CHIR99021, RA and FGF7 for regenerating IPCs. These data showed that the 2-stage guided differentiation of iPSCs into IPCs in CHIR99021-SLNs-GDF8-AHA-PAAM ICCSs can be promising in ameliorating insulin secretion by β-cell regeneration.

Fabrication of Bioactive Inverted Colloidal Crystal Scaffolds Using Expanded Polystyrene Beads.

Inverted colloidal crystal (ICC) hydrogel scaffolds have emerged as a new class of three-dimensional cell culture matrix that represents a unique opportunity to reproduce lymphoid tissue microenvironments. ICC geometry promotes the formation of stromal cell networks and their interaction with hematopoietic cells, a core cellular process in lymphoid tissues. When subdermally implanted, ICC hydrogel scaffolds direct unique foreign body responses to form a vascularized stromal tissue with prolonged attraction of hematopoietic cells, which together resemble lymphoid tissue microenvironments. While conceptually simple, fabrication of ICC hydrogel scaffold requires multiple steps and laborious handling of delicate materials. Here, we introduce a facile route for ICC hydrogel scaffold fabrication using expanded polystyrene (EPS) beads. EPS beads shrink and fuse in a tunable manner under pressurized thermal conditions, which serves as colloidal crystal templates for ICC scaffold fabrication. Inclusion of collagen in the precursor solution greatly simplified preparation of bioactive hydrogel scaffolds. The resultant EPS-templated bioactive ICC hydrogel scaffolds demonstrate characteristic features required for lymphoid tissue modeling in both in vitro and in vivo settings. We envision that the presented method will facilitate widespread implementation of ICC hydrogel scaffolds for lymphoid tissue engineering and other emerging applications. Impact statement Inverted colloidal crystal (ICC) hydrogel scaffolds have emerged as a new class of three-dimensional cell culture matrix that represents a unique opportunity for lymphoid tissue modeling and other emerging novel bioengineering applications. While conceptually simple, fabrication of the ICC hydrogel scaffold requires multiple steps and laborious handling of delicate materials with highly toxic chemicals. The presented method for ICC hydrogel scaffold fabrication using expanded polystyrene (EPS) beads is simple, cost-effective, and involves less toxic chemicals than conventional methods, while retaining comparable biological significance. We envision that EPS bead-based hydrogel scaffold fabrication will greatly facilitate the widespread implementation of ICC hydrogel scaffolds and their practical applications.

Lymphoid Tissue Engineering: Thermoresponsive Inverted Colloidal Crystal Hydrogel Scaffolds for Lymphoid Tissue Engineering (Adv. Healthcare Mater. 6/2020)

Lymphoid Tissue Engineering: Thermoresponsive Inverted Colloidal Crystal Hydrogel Scaffolds for Lymphoid Tissue Engineering (Adv. Healthcare Mater. 6/2020)

Thermoresponsive Inverted Colloidal Crystal Hydrogel Scaffolds for Lymphoid Tissue Engineering.

Inverted colloidal crystal (ICC) hydrogel scaffolds represent unique opportunities in modeling lymphoid tissues and expanding hematopoietic-lymphoid cells. Fully interconnected spherical pore arrays direct the formation of stromal networks and facilitate interactions between stroma and hematopoietic-lymphoid cells. However, due to the intricate architecture of these materials, release of expanded cells is restricted and requires mechanical disruption or chemical dissolution of the hydrogel scaffold. One potent biomaterials strategy to release pore-entrapped hematopoietic-lymphoid cells without breaking the scaffolds apart is to transiently increase the dimensions of these materials using stimuli-responsive polymers. Having this mindset, thermoresponsive ICC scaffolds that undergo rapid (<1 min) and substantial (>300%) diameter change over a physiological temperature range (4-37 °C) by using poly(N-isopropylacrylamide) (PNIPAM) with nanogel crosslinkers is developed. For a proof-of-concept study, the stromal niche by creating osteospheroids, aggregates of osteoblasts, and bone chips is first replicated, and subsequently Nalm-6 model hematopoietic-lymphoid cells are introduced. A sixfold increase in cell count is harvested when ICC hydrogel scaffolds are expanded without termination of the established 3D stromal cell culture. It is envisioned that thermoresponsive ICC hydrogel scaffolds will enable for scalable and sustainable ex vivo expansion of hematopoietic-lymphoid cells.

Porcine hepatocytes culture on biofunctionalized 3D inverted colloidal crystal scaffolds as anin vitromodel for predicting drug hepatotoxicity

As drug-induced hepatotoxicity represents one of the most common causes of drug failure, three-dimensional (3D) in vitro liver platforms represent a fantastic toolbox to predict drug toxicity and thus reduce in vivo animal studies and lessen drug attrition rates. The aim of this study is to establish a functional porcine hepatocyte culture using a biofunctionalized 3D inverted colloidal crystal (ICC) hydrogel platform. The performances of non-adhesive bare poly(ethylene glycol)diacrylate (PEGDA) ICCs and PEGDA ICCs coated with either collagen type I or fibronectin have been investigated. Porcine hepatocytes viability, morphology, hepatic-specific functions and patterns of gene expression have been evaluated over a period of two weeks in culture to test diclofenac, a well-known hepatotoxic drug. Interestingly, cells in the fibronectin-functionalized scaffold exhibit different aggregation patterns and maintain better liver-specific function than those in bare ICCs and in collagen functionalized scaffold. We concluded that the 3D cell culture environment and the presence of extracellular matrix (ECM) proteins, especially fibronectin, facilitate hepatocyte viability and maintenance of the liver-specific phenotype in vitro, and enable us to predict hepatotoxicity.

Human iPS derived progenitors bioengineered into liver organoids using an inverted colloidal crystal poly (ethylene glycol) scaffold

Generation of human organoids from induced pluripotent stem cells (iPSCs) offers exciting possibilities for developmental biology, disease modelling and cell therapy. Significant advances towards those goals have been hampered by dependence on animal derived matrices (e.g. Matrigel), immortalized cell lines and resultant structures that are difficult to control or scale. To address these challenges, we aimed to develop a fully defined liver organoid platform using inverted colloid crystal (ICC) whose 3-dimensional mechanical properties could be engineered to recapitulate the extracellular niche sensed by hepatic progenitors during human development. iPSC derived hepatic progenitors (IH) formed organoids most optimally in ICC scaffolds constructed with 140 μm diameter pores coated with type I collagen in a two-step process mimicking liver bud formation. The resultant organoids were closer to adult tissue, compared to 2D and 3D controls, with respect to morphology, gene expression, protein secretion, drug metabolism and viral infection and could integrate, vascularise and function following implantation into livers of immune-deficient mice. Preliminary interrogation of the underpinning mechanisms highlighted the importance of TGFβ and hedgehog signalling pathways. The combination of functional relevance with tuneable mechanical properties leads us to propose this bioengineered platform to be ideally suited for a range of future mechanistic and clinical organoid related applications.

Colloidal templating of highly ordered gelatin methacryloyl-based hydrogel platforms for three-dimensional tissue analogues

Three-dimensional, protein-based hydrogel scaffolds that successfully mimic in vivo extracellular matrix microenvironments are desirable for tissue engineering and regenerative medicine applications, and can provide highly capable in vitro tissue analogues. However, the fabrication of protein-based scaffolds with uniform porosity, thin walls and durable mechanical properties remains a challenging prospect that might be overcome by integrating advances in microfabrication and protein functionalization. Towards this goal, herein, we report the successful fabrication of a highly ordered, gelatin-based inverted colloidal crystal (ICC) hydrogel platform that is robust and supports high levels of cell function. In particular, the utilization of colloidal templating microfabrication strategies together with highly substituted, photocrosslinkable gelatin methacryloyl (GelMA) allowed us to fabricate protein-based three-dimensional scaffolds with uniform pore interconnectivity, structural stability and tailorable degradation properties. The resulting GelMA ICC scaffolds provided cell attachment sites and promoted intercellular interaction of hepatocytes, which resulted in improved cell function compared to a flat 2D system. The results demonstrate the potential of GelMA ICC scaffolds to become an effective tissue engineering platform for drug screening and regenerative medicine.

Neuroregeneration of Induced Pluripotent Stem Cells in Polyacrylamide-Chitosan Inverted Colloidal Crystal Scaffolds with Poly(lactide-co-glycolide) Nanoparticles and Transactivator of Transcription von Hippel-Lindau Peptide.

Polyacrylamide (PAAM) and chitosan were fabricated by inverted colloidal crystal (ICC) method for scaffolds comprising regular pores. The hybrid PAAM-chitosan ICC scaffolds were grafted with poly(lactide-co-glycolide) (PLGA) nanoparticles (NPs) for a rougher pore surface and grafted with transactivator of transcription von Hippel-Lindau (TATVHL) peptide for a better differentiation of induced pluripotent stem (iPS) cells toward neural lineage. By scanning electron microscopy, we found that iPS cells cultured in PAAM-chitosan ICC scaffolds with PLGA NPs at 1.0 mg/mL and TATVHL peptide at 15 μg/mL elongated the axonal length to 15 μm. A combination of PLGA NPs and TATVHL peptide favored the adhesion of iPS cells, reduced the embryonic phenotype after cultivation, and guided the production of βIII tubulin-positive cells in PAAM-chitosan ICC scaffolds. In addition to the differentiation toward neurite-like cells, an increase in the content of TATVHL peptide in PAAM-chitosan ICC scaffolds inhibited the differentiation of iPS cells toward astrocytes. ICC scaffolds composed of PAAM, chitosan, PLGA NPs, and TATVHL peptide can be an efficacious matrix to differentiate iPS cells toward neurons and retard the glial formation for nerve regeneration.

Chitosan Inverted Colloidal Crystal scaffolds: Influence of molecular weight on structural stability

Chitosan with three different molecular weights (538±48, 229±45 and 13±3kDa) was used to develop biodegradable Inverted Colloidal Crystal (ICC) scaffolds with uniform pore size and interconnected pore network. Mass loss and compression modulus were analyzed after hydrolytic degradation in order to understand the influence of molecular weight on structural and mechanical degradation of chitosan ICC structures. Results show that medium molecular weight chitosan (229±45kDa) retains ICC structure and compression modulus for an extended period (4weeks) and is therefore the preferred one for the production of ICC for soft tissue engineering.

ECM proteins in a microporous scaffold influence hepatocyte morphology, function, and gene expression.

It is well known that a three-dimensional (3D) culture environment and the presence of extracellular matrix (ECM) proteins facilitate hepatocyte viability and maintenance of the liver-specific phenotype in vitro. However, it is not clear whether specific ECM components such as collagen or fibronectin differentially regulate such processes, especially in 3D scaffolds. In this study, a series of ECM-functionalized inverted colloidal crystal (ICC) microporous scaffolds were fabricated and their influence on Huh-7.5 cell proliferation, morphology, hepatic-specific functions, and patterns of gene expression were compared. Both collagen and fibronectin promoted albumin production and liver-specific gene expression of Huh-7.5 cells, compared with the bare ICC scaffold. Interestingly, cells in the fibronectin-functionalized scaffold exhibited different aggregation patterns to those in the collagen-functionalized scaffold, a variation that could be related to the distinct mRNA expression levels of cell adhesion-related genes. Based on these results, we can conclude that different ECM proteins, such as fibronectin and collagen, indeed play distinct roles in the phenotypic regulation of cells cultured in a 3D environment.

A simple sol-gel route to the construction of hydroxyapatite inverted colloidal crystals for bone tissue engineering

Hydroxyapatite (HAp) scaffolds with uniform pore size and interconnected pore network were constructed based on the inverted colloidal crystal (ICC) geometry and a simple sol-gel formulation. Monodisperse polystyrene microspheres were self-assembled and annealed into a hexagonal close packed structure. HAp sol-gel was infiltrated in this template followed by thermal treatment for simultaneous HAp matrix sintering and polymeric colloidal crystal calcination. The resultant ICC scaffolds exhibit an ordered architecture that was able to offer a favorable environment for human osteoblasts adhesion and proliferation, an essential feature for bone ingrowth in tissue engineering applications.

Fabrication of Inverted Colloidal Crystal Poly(ethylene glycol) Scaffold: A Three-dimensional Cell Culture Platform for Liver Tissue Engineering.

The ability to maintain hepatocyte function in vitro, for the purpose of testing xenobiotics' cytotoxicity, studying virus infection and developing drugs targeted at the liver, requires a platform in which cells receive proper biochemical and mechanical cues. Recent liver tissue engineering systems have employed three-dimensional (3D) scaffolds composed of synthetic or natural hydrogels, given their high water retention and their ability to provide the mechanical stimuli needed by the cells. There has been growing interest in the inverted colloidal crystal (ICC) scaffold, a recent development, which allows high spatial organization, homotypic and heterotypic cell interaction, as well as cell-extracellular matrix (ECM) interaction. Herein, we describe a protocol to fabricate the ICC scaffold using poly (ethylene glycol) diacrylate (PEGDA) and the particle leaching method. Briefly, a lattice is made from microsphere particles, after which a pre-polymer solution is added, properly polymerized, and the particles are then removed, or leached, using an organic solvent (e.g., tetrahydrofuran). The dissolution of the lattice results in a highly porous scaffold with controlled pore sizes and interconnectivities that allow media to reach cells more easily. This unique structure allows high surface area for the cells to adhere to as well as easy communication between pores, and the ability to coat the PEGDA ICC scaffold with proteins also shows a marked effect on cell performance. We analyze the morphology of the scaffold as well as the hepatocarcinoma cell (Huh-7.5) behavior in terms of viability and function to explore the effect of ICC structure and ECM coatings. Overall, this paper provides a detailed protocol of an emerging scaffold that has wide applications in tissue engineering, especially liver tissue engineering.

Interpenetrating polymer network hydrogel scaffolds for artificial cornea periphery.

Three-dimensional scaffolds based on inverted colloidal crystals (ICCs) were fabricated from sequentially polymerized interpenetrating polymer network (IPN) hydrogels of poly(ethyleneglycol) and poly(acrylic acid). This high-strength, high-water-content IPN hydrogel may be suitable for use in an artificial cornea application. Development of a highly porous, biointegrable region at the periphery of the artificial cornea device is critical to long-term retention of the implant. The ICC fabrication technique produced scaffolds with well-controlled, tunable pore and channel dimensions. When surface functionalized with extracellular matrix proteins, corneal fibroblasts were successfully cultured on IPN hydrogel scaffolds, demonstrating the feasibility of these gels as materials for the artificial cornea porous periphery. Porous hydrogels with and without cells were visualized non-invasively in the hydrated state using variable-pressure scanning electron microscopy.

MO‐G‐BRF‐07: Anomalously Fast Diffusion of Carbon Nanotubes Carriers in 3D Tissue Model

Purpose:We aim to investigate and understand diffusion process of carbon nanotubes (CNTs) and other nanoscale particles in tissue and organs.Methods:In this research, we utilized a 3D model tissue of hepatocellular carcinoma (HCC)cultured in inverted colloidal crystal (ICC) scaffolds to compare the diffusivity of CNTs with small molecules such as Rhodamine and FITC in vitro, and further investigated the transportation of CNTs with and without targeting ligand, TGFβ1. The real‐time permeation profiles of CNTs in HCC tissue model with high temporal and spatial resolution was demonstrated by using standard confocal microscopy. Quantitative analysis of the diffusion process in 3D was carried out using luminescence intensity in a series of Z‐stack images obtained for different time points of the diffusion process after initial addition of CNTs or small molecules to the cell culture and the image data was analyzed by software ImageJ and Mathematica.Results:CNTs display diffusion rate in model tissues substantially faster than small molecules of the similar charge such as FITC, and the diffusion rate of CNTs are significantly enhanced with targeting ligand, TGFβ1.Conclusion:In terms of the advantages of in‐vitro model, we were able to have access to measuring the rate of CNT penetration at designed conditions with variable parameters. And the findings by using this model, changed our understanding about advantages of CNTs as nanoscale drug carriers and provides design principles for making new drug carriers for both treatment and diagnostics. Additionally the fast diffusion opens the discussion of the best possible drug carriers to reach deep parts of cancerous tissues, which is often a prerequisite for successful cancer treatment.This work was supported by the Center for Photonic and Multiscale Nanomaterials funded by National Science Foundation Materials Research Science and Engineering Center program DMR 1120923. The work was also partially supported by NSF grant ECS‐0601345; EFRI‐BSBA 0938019; CBET 0933384; CBET 0932823; CBET 1036672, AFOSR MURI 444286‐P061716 and NIH 1R21CA121841‐01A2.

An Overview of Inverted Colloidal Crystal Systems for Tissue Engineering

Scaffolding is at the heart of tissue engineering but the number of techniques available for turning biomaterials into scaffolds displaying the features required for a tissue engineering application is somewhat limited. Inverted colloidal crystals (ICCs) are inverse replicas of an ordered array of monodisperse colloidal particles, which organize themselves in packed long-range crystals. The literature on ICC systems has grown enormously in the past 20 years, driven by the need to find organized macroporous structures. Although replicating the structure of packed colloidal crystals (CCs) into solid structures has produced a wide range of advanced materials (e.g., photonic crystals, catalysts, and membranes) only in recent years have ICCs been evaluated as devices for medical/pharmaceutical and tissue engineering applications. The geometry, size, pore density, and interconnectivity are features of the scaffold that strongly affect the cell environment with consequences on cell adhesion, proliferation, and differentiation. ICC scaffolds are highly geometrically ordered structures with increased porosity and connectivity, which enhances oxygen and nutrient diffusion, providing optimum cellular development. In comparison to other types of scaffolds, ICCs have three major unique features: the isotropic three-dimensional environment, comprising highly uniform and size-controllable pores, and the presence of windows connecting adjacent pores. Thus far, this is the only technique that guarantees these features with a long-range order, between a few nanometers and thousands of micrometers. In this review, we present the current development status of ICC scaffolds for tissue engineering applications.

Replication of Bone Marrow Differentiation Niche: Comparative Evaluation of Different Three‐Dimensional Matrices

The comparative evaluation of different 3D matrices-Matrigel, Puramatrix, and inverted colloidal crystal (ICC) scaffolds-provides a perspective for studying the pathology and potential cures for many blood and bone marrow diseases, and further proves the significance of 3D cultures with direct cell-cell contacts for in vitro mimicry of the human stem cell niche.

Accelerated nerve regeneration using induced pluripotent stem cells in chitin–chitosan–gelatin scaffolds with inverted colloidal crystal geometry

Polymeric biomaterials with regular pore structure can generate distinctive properties in various biomedical applications. This study presents scaffolds comprising chitin, chitosan, and gelatin with pore geometry of inverted colloidal crystals (ICC) for guiding differentiation of induced pluripotent stem (iPS) cells toward neurons. Chitin–chitosan–gelatin ICC scaffolds were fabricated by self-assembly, crosslinking, infiltration, dehydration, and particle leaching. The results revealed that ethanol as a dispersion medium yielded a higher regularity of colloidal template than ethylene glycol. For the adhesion of iPS cells, freeform constructs were more efficient than ICC constructs. The reverse was true for the viability of iPS cells. The quantity of stage-specific embryonic surface antigen-1 in cultured freeform construct was larger than that in cultured ICC construct, indicating that the former preserved more phenotypic characteristics of iPS cells than the latter. Moreover, β III tubulin-identified region in ICC construct was larger than that in freeform construct, demonstrating that the differentiation of iPS cells toward neurons in ICC construct was faster than that in freeform construct. ICC topography in chitin–chitosan–gelatin scaffolds can accelerate neuronal differentiation of iPS cells.

TATVHL peptide-grafted alginate/poly(γ-glutamic acid) scaffolds with inverted colloidal crystal topology for neuronal differentiation of iPS cells

The neuronal differentiation of induced pluripotent stem (iPS) cells in scaffolding biomaterials is an emerging issue in nervous regeneration and repair. This study presents the production of neuron-lineage cells from iPS cells in inverted colloidal crystal (ICC) scaffolds comprising alginate, poly(γ-glutamic acid) (γ-PGA), and TATVHL peptide. The ability of iPS cells to differentiate toward neurons in the constructs was demonstrated by flow-cytometeric sorting and immunochemical staining. The results revealed that hexagonally arrayed microspheres molded alginate/γ-PGA hydrogel into ICC topology with adequate interconnected pores. An increase in the quantity of surface TATVHL peptide enhanced the atomic ratio of nitrogen and the adhesion efficiency of iPS cells in constructs. However, the effect of TATVHL peptide on the viability of iPS cells was insignificant. The adhesion and viability of iPS cells in ICC constructs was higher than those in freeform ones. TATVHL peptide raised the percentage of β III tubulin-identified cells differentiating from iPS cells, indicating that TATVHL peptide stimulated the neuronal development in alginate/γ-PGA ICC constructs. TATVHL peptide-grafted alginate/γ-PGA ICC scaffolds can be promising for establishing nerve tissue from iPS cells.

Inverted colloidal crystal scaffolds with induced pluripotent stem cells for nerve tissue engineering

The development of biomaterials for regenerating neurons from induced pluripotent stem (iPS) cells is crucial to the potential therapy for traumatic injury to nervous system. This study aims to guide differentiation of iPS cells into neuron-lineage cells in inverted colloidal crystal (ICC) scaffolds containing alginate, poly(γ-glutamic acid), and surface CSRARKQAASIKVAVSADR (peptide). The differentiation of iPS cells in ICC constructs was characterized by staining of embryonic and neuronal markers. The results indicated that hexagonal crystals of polystyrene microspheres shaped hydrogels into ICC scaffolds with interconnected pores. CSRARKQAASIKVAVSADR slightly enhanced the adhesion of iPS cells in ICC constructs and yielded no variation in the viability of iPS cells. Cultured ICC constructs with CSRARKQAASIKVAVSADR reduced the expression of stage-specific embryonic surface antigen-1 and raised the expression of β III tubulin of differentiating iPS cells. The induction with CSRARKQAASIKVAVSADR in ICC topography can improve the differentiation of iPS cells toward neurons for nerve tissue engineering.