Improving fluorescence imaging of biological cells on biomedical polymers

Jaafar, Israd Hakim; LeBlon, Courtney; Wei, Ming-Tzo; Ou-Yang, Daniel; Coulter, John P.; Jedlicka, Sabrina S.

doi:10.1016/j.actbio.2010.12.007

Cited by 40 publications

(34 citation statements)

References 35 publications

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“…Thus though, optical detection of NFC hydrogel with light microscopy may be limited due to light scattering. Lack of autofluorescence allows fluorescence spectroscopy based imaging with low background unlike with other biomaterial scaffolds [28].…”

Section: Optical Propertiesmentioning

confidence: 99%

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Bhattacharya

Malinen

Laurén

et al. 2012

Journal of Controlled Release

368

290

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Over the recent years, various materials have been introduced as potential 3D cell culture scaffolds. These include protein extracts, peptide amphiphiles, and synthetic polymers. Hydrogel scaffolds without human or animal borne components or added bioactive components are preferred from the immunological point of view. Here we demonstrate that native nanofibrillar cellulose (NFC) hydrogels derived from the abundant plant sources provide the desired functionalities. We show 1) rheological properties that allow formation of a 3D scaffold in-situ after facile injection, 2) cellular biocompatibility without added growth factors, 3) cellular polarization, and 4) differentiation of human hepatic cell lines HepaRG and HepG2. At high shear stress, the aqueous NFC has small viscosity that supports injectability, whereas at low shear stress conditions the material is converted to an elastic gel. Due to the inherent biocompatibility without any additives, we conclude that NFC generates a feasible and sustained microenvironment for 3D cell culture for potential applications, such as drug and chemical testing, tissue engineering, and cell therapy.

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Section: Optical Propertiesmentioning

confidence: 99%

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Bhattacharya

Malinen

Laurén

et al. 2012

Journal of Controlled Release

368

290

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“…The PEMs coated specimens with chitosan as the outer layer show similar affinity of cell adhesion as bare metal substrates, which is in coherence with the cell viability result. Polymeric materials are known to hinder fluorescence imaging due to either autofluorescence or light scattering property of polymers [34]; thus, it is not possible to clearly identify the cells on PEMs coated specimens with PGA as the finishing layer in this experiment. A more direct method to observe the cell adhesion and affinity on a material is to observe cell attachment and morphology using fluorescence microscopy.…”

Section: Biocompatibilitymentioning

confidence: 77%

Preparation of Chitosan/Poly‐γ‐Glutamic Acid Polyelectrolyte Multilayers on Biomedical Metals for Local Antibiotic Delivery

Liu

Wang

et al. 2017

Metals

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Abstract:Polyelectrolyte multilayer assembly is one of the most widely applied biomaterial coatings for applications from surface modification, drug delivery, tissue engineering to biomimetic extracellular environment. In this research, we propose a simple layer-wise spin coating technique to prepare chitosan/poly-γ-glutamic acid (C/PGA) polyelectrolyte multilayers (PEMs) on two different biomedical metals, 316L stainless steel (316LSS) and titanium alloy (Ti6Al4V). The multilayer coating was fabricated using oppositely charged chitosan and poly-γ-glutamic acid to deposit a total of 10, 20, or 30 multilayered films. Afterward, tetracycline was loaded by soaking the coated metals for 12 hours. The microstructure, mechanical properties, biocompatibility and drug release rate were investigated by scanning electron microscopy, contact angle measurement, MG63 cell viability and inhibition of Escherichia coli (E. coli) growth. Lastly, MG63 cell attachment was detected by fluorescence microscopy after staining with Hoechst 33258. This coating technique can prepare a layer of 2.2-6.9 µm C/PGA PEMs favoring cell attachment and growth. Moreover, tetracycline was released from C/PGA PEMs and inhibited the growth of E. coli. The results suggest that C/PGA PEMs provide a useful platform for modulating the micro-environment for better cell adhesion and antibiotic delivery, which hold great potential for surface modification and drug loading for biomimetic materials.

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“…Some of the original polymer nanolattices were used for fluorescence studies, which revealed the need to treat the polymer nanolattices with Sudan Black to suppress autofluorescence according to the protocol developed by Jaafar et al 30 (supplementary information, Fig. S1).…”

Section: Methodsmentioning

confidence: 99%

Three-dimensional nano-architected scaffolds with tunable stiffness for efficient bone tissue growth

Maggi

Greer

2017

Acta Biomaterialia

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The precise mechanisms that lead to orthopedic implant failure are not well understood; it is believed that the micromechanical environment at the bone-implant interface regulates structural stability of an implant. In this work, we seek to understand how the 3D mechanical environment of an implant affects bone formation during early osteointegration. We employed two-photon lithography (TPL) direct laser writing to fabricate 3-dimensional rigid polymer scaffolds with tetrakaidecahedral periodic geometry, herewith referred to as nanolattices, whose strut dimensions were on the same order as osteoblasts’ focal adhesions (~2μm) and pore sizes on the order of cell size, ~10μm. Some of these nanolattices were subsequently coated with thin conformal layers of Ti or W, and a final outer layer of 18nm-thick TiO2 was deposited on all samples to ensure biocompatibility. Nanomechanical experiments on each type of nanolattice revealed the range of stiffnesses of 0.7–100 MPa. Osteoblast-like cells (SAOS-2) were seeded on each nanolattice, and their mechanosensitve response was explored by tracking mineral secretions and intracellular f-actin and vinculin concentrations after 2, 8 and 12 days of cell culture in mineralization media. Experiments revealed that the most compliant nanolattices had ~20% more intracellular f-actin and ~40% more Ca and P secreted onto them than the stiffer nanolattices, where such cellular response was virtually indistinguishable. We constructed a simple phenomenological model that appears to capture the observed relation between scaffold stiffness and f-actin concentration. This model predicts a range of optimal scaffold stiffnesses for maximum f-actin concentration, which appears to be directly correlated with osteoblast-driven mineral deposition. This work suggests that three-dimensional scaffolds with titania-coated surfaces may provide an optimal microenvironment for cell growth when their stiffness is similar to that of cartilage (~0.5–3MPa). These findings help provide a greater understanding of osteoblast mechanosensitivity and may have profound implications in developing more effective and safer bone prostheses.

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Improving fluorescence imaging of biological cells on biomedical polymers

Cited by 40 publications

References 35 publications

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Nanofibrillar cellulose hydrogel promotes three-dimensional liver cell culture

Preparation of Chitosan/Poly‐γ‐Glutamic Acid Polyelectrolyte Multilayers on Biomedical Metals for Local Antibiotic Delivery

Three-dimensional nano-architected scaffolds with tunable stiffness for efficient bone tissue growth

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