Measuring and mapping biophysical properties and their influence on mesenchymal stem cell fate

Mesenchymal stem cells (MSCs) as adherent stem cells are being increasingly used as a therapeutic cell source, thus developing a better understanding of how to control MSC fate choice is an increasingly important task. Unlike the well known chemical cues, the biophysical properties of the surrounding extracellular environment, such as the appropriate spatial display of adhesion sites and environment stiffness should be clarified. The atomic force microscopy (AFM)-based technique called force spectroscopy mapping (FSM) was applied for examination biophysical properties such as elasticity and adhesion site size and spacing. Using the indentation, the spatial changes in the mechanics of poly(vinyl pyrrolidone) (PVP) and poly(acrylamide) (PAam) hydrogels were detected. PVP hydrogels had very heterogeneous elastic moduli as a function of hydrogel position as well as the amount of crosslinker. PAam hydrogels were a much more homogeneous substrate, showing little spatial variation in moduli. Coupled with chemical-functionalized probe, the technique of chemical FSM (CFSM) was shown the capability of recognizing sub-micron adhesive sites from probe retraction studies. By functionalizing the probe to recognize the charged surface of copolymers, the spatial display of adhesion sites in diblock copolymer foams was investiaged. Prepared by high internal phase emulsion templating using amphiphilic copolymers of polystyrene- block-polyacrylic acid and polystyrene-block-polyethylene oxide, the porous foams have been synthesized and characterized. With nanoscopic domains of cell inert and active chemistries mimicking native matrix, the nanodomains of adhesive sites were detected by CFSM. Protein adsorption on surfaces was examined in spatially macro-, micro-, and nanoscopic level. MSCs from different cell sources of human-embryonic stem-mesoderm progenitors (hES-MPs) and human bone marrow derived-mesenchymal stem cells (hBMSCs) were examined by quantitative PCR to assess their expression of myogenic, adipogenic, and osteogenic genotypes as a result of their interaction with the foams of varying composition. Interestingly, without the induction media, hBMSCs expressed adipogenic genes and proteins on 'patchy' matrices where adhesive nano-domains were present. More interestingly, and their expression depends on stem cell origin: marrow-derived and mesenchymal progenitor cells exhibit fundamentally different differentiation patterns, adipo- and osteogenic, respectively. Together these data for the first time implicate adhesion as a complex regulator of cell fat

Measuring and mapping biophysical properties and their influence on mesenchymal stem cell fate

Mesenchymal stem cells (MSCs) as adherent stem cells are being increasingly used as a therapeutic cell source, thus developing a better understanding of how to control MSC fate choice is an increasingly important task. Unlike the well known chemical cues, the biophysical properties of the surrounding extracellular environment, such as the appropriate spatial display of adhesion sites and environment stiffness should be clarified. The atomic force microscopy (AFM)-based technique called force spectroscopy mapping (FSM) was applied for examination biophysical properties such as elasticity and adhesion site size and spacing. Using the indentation, the spatial changes in the mechanics of poly(vinyl pyrrolidone) (PVP) and poly(acrylamide) (PAam) hydrogels were detected. PVP hydrogels had very heterogeneous elastic moduli as a function of hydrogel position as well as the amount of crosslinker. PAam hydrogels were a much more homogeneous substrate, showing little spatial variation in moduli. Coupled with chemical-functionalized probe, the technique of chemical FSM (CFSM) was shown the capability of recognizing sub-micron adhesive sites from probe retraction studies. By functionalizing the probe to recognize the charged surface of copolymers, the spatial display of adhesion sites in diblock copolymer foams was investiaged. Prepared by high internal phase emulsion templating using amphiphilic copolymers of polystyrene- block-polyacrylic acid and polystyrene-block-polyethylene oxide, the porous foams have been synthesized and characterized. With nanoscopic domains of cell inert and active chemistries mimicking native matrix, the nanodomains of adhesive sites were detected by CFSM. Protein adsorption on surfaces was examined in spatially macro-, micro-, and nanoscopic level. MSCs from different cell sources of human-embryonic stem-mesoderm progenitors (hES-MPs) and human bone marrow derived-mesenchymal stem cells (hBMSCs) were examined by quantitative PCR to assess their expression of myogenic, adipogenic, and osteogenic genotypes as a result of their interaction with the foams of varying composition. Interestingly, without the induction media, hBMSCs expressed adipogenic genes and proteins on 'patchy' matrices where adhesive nano-domains were present. More interestingly, and their expression depends on stem cell origin: marrow-derived and mesenchymal progenitor cells exhibit fundamentally different differentiation patterns, adipo- and osteogenic, respectively. Together these data for the first time implicate adhesion as a complex regulator of cell fat

Hemagglutination Detection with Paper–Plastic Hybrid Passive Microfluidic Chip

Hemagglutination is a critical reaction that occurs when antigens expressed on red blood cells (RBCs) react with the antibodies used for blood typing. Even though blood typing devices have been introduced to the market, they continue to face several limitations in terms of observation by the eye alone, blood manipulation difficulties, and the need for large-scale equipment, particularly process automated machines. Thus, this study aimed to design, fabricate, and test a novel hybrid passive microfluidic chip made of filter paper and polymer using a cost-effective xurography manufacturing technique. This chip is referred to as the microfluidic paper–plastic hybrid passive device (PPHD). A passive PPHD does not require external sources, such as a syringe pump. It is composed of a paper-based component that contains dried antibodies within its porous paper and a polymer component that serves as the detection zone. A single blood sample was injected into the chip’s inlet, and classification was determined using the mean intensity image. The results indicated that embedded antibodies were capable of causing RBC agglutination without a saline washing step and that the results could be classified as obviously agglutination or nonagglutination for blood typing using both the naked eye and a mean intensity image. As a proof-of-concept, this study demonstrated efficiency in quantitative hemagglutination measurement within a passive PPHD for blood typing, which could be used to simplify blood biomarker analysis

Cell Instructive Microporous Scaffolds Through Interface Engineering

The design of novel biomaterials for regenerative medicine requires incorporation of well-defined physical and chemical properties that mimic the native extracellular matrix (ECM). Here, we report the synthesis and characterisation of porous foams prepared by high internal phase emulsion (HIPE) templating using amphiphilic copolymers that act as surfactants during the HIPE process. We combine different copolymers exploiting oil-water interface confined phase separation to engineer the surface topology of foam pores with nanoscopic domains of cell inert and active chemistries mimicking native matrix. We further demonstrate how proteins and hMSCs adhere in a domain specific manner

Cell Instructive Microporous Scaffolds through Interface Engineering

The design of novel biomaterials for regenerative medicine requires incorporation of well-defined physical and chemical properties that mimic the native extracellular matrix (ECM). Here, we report the synthesis and characterization of porous foams prepared by high internal phase emulsion (HIPE) templating using amphiphilic copolymers that act as surfactants during the HIPE process. We combine different copolymers exploiting oil–water interface confined phase separation to engineer the surface topology of foam pores with nanoscopic domains of cell inert and active chemistries mimicking native matrix. We further demonstrate how proteins and hMSCs adhere in a domain specific manner

Cell Instructive Microporous Scaffolds through Interface Engineering

The design of novel biomaterials for regenerative medicine requires incorporation of well-defined physical and chemical properties that mimic the native extracellular matrix (ECM). Here, we report the synthesis and characterization of porous foams prepared by high internal phase emulsion (HIPE) templating using amphiphilic copolymers that act as surfactants during the HIPE process. We combine different copolymers exploiting oil–water interface confined phase separation to engineer the surface topology of foam pores with nanoscopic domains of cell inert and active chemistries mimicking native matrix. We further demonstrate how proteins and hMSCs adhere in a domain specific manner

3D surface topology guides stem cell adhesion and differentiation.

Polymerized high internal phase emulsion (polyHIPE) foams are extremely versatile materials for investigating cell-substrate interactions in vitro. Foam morphologies can be controlled by polymerization conditions to result in either open or closed pore structures with different levels of connectivity, consequently enabling the comparison between 2D and 3D matrices using the same substrate with identical surface chemistry conditions. Additionally, here we achieve the control of pore surface topology (i.e. how different ligands are clustered together) using amphiphilic block copolymers as emulsion stabilizers. We demonstrate that adhesion of human mesenchymal progenitor (hES-MP) cells cultured on polyHIPE foams is dependent on foam surface topology and chemistry but is independent of porosity and interconnectivity. We also demonstrate that the interconnectivity, architecture and surface topology of the foams has an effect on the osteogenic differentiation potential of hES-MP cells. Together these data demonstrate that the adhesive heterogeneity of a 3D scaffold could regulate not only mesenchymal stem cell attachment but also cell behavior in the absence of soluble growth factors