scholarly journals Self-assembly study of type I collagen extracted from male Wistar Hannover rat tail tendons

2020 ◽  
Vol 24 (1) ◽  
Author(s):  
Jeimmy González-Masís ◽  
Jorge M. Cubero-Sesin ◽  
Simón Guerrero ◽  
Sara González-Camacho ◽  
Yendry Regina Corrales-Ureña ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Self Assembly ◽  
Type I Collagen ◽  
Ph Effect ◽  
The Self ◽  
Titration Calorimetry ◽  
Type I ◽  
Self Assembling ◽  
Functional Features ◽  
Rat Tail

Abstract Background Collagen, the most abundant protein in the animal kingdom, represents a promising biomaterial for regenerative medicine applications due to its structural diversity and self-assembling complexity. Despite collagen’s widely known structural and functional features, the thermodynamics behind its fibrillogenic self-assembling process is still to be fully understood. In this work we report on a series of spectroscopic, mechanical, morphological and thermodynamic characterizations of high purity type I collagen (with a D-pattern of 65 nm) extracted from Wistar Hannover rat tail. Our herein reported results can be of help to elucidate differences in self-assembly states of proteins using ITC to improve the design of energy responsive and dynamic materials for applications in tissue engineering and regenerative medicine. Methods Herein we report the systematic study on the self-assembling fibrillogenesis mechanism of type I collagen, we provide morphological and thermodynamic evidence associated to different self-assembly events using ITC titrations. We provide thorough characterization of the effect of pH, effect of salts and protein conformation on self-assembled collagen samples via several complementary biophysical techniques, including circular dichroism (CD), Fourier Transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), atomic force microscopy (AFM), scanning electron microscopy (SEM), dynamic mechanical thermal analysis (DMTA) and thermogravimetric analysis (TGA). Results Emphasis was made on the use of isothermal titration calorimetry (ITC) for the thermodynamic monitoring of fibrillogenesis stages of the protein. An overall self-assembly enthalpy value of 3.27 ± 0.85 J/mol was found. Different stages of the self-assembly mechanism were identified, initial stages take place at pH values lower than the protein isoelectric point (pI), however, higher energy release events were recorded at collagen’s pI. Denatured collagen employed as a control exhibited higher energy absorption at its pI, suggesting different energy exchange mechanisms as a consequence of different aggregation routes. Graphical abstract

scholarly journals Self-assembly study of Type I collagen extracted from male Wistar Hannover rat tail tendons

2020 ◽  
Author(s):  
Jeimmy C. González-Masís ◽  
Jorge M. Cubero-Sesin ◽  
Simón Guerrero ◽  
Sara González-Camacho ◽  
Yendry Regina Corrales-Ureña ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Self Assembly ◽  
Type I Collagen ◽  
Ph Effect ◽  
The Self ◽  
Titration Calorimetry ◽  
Type I ◽  
Self Assembling ◽  
Functional Features ◽  
Rat Tail

Abstract BackgroundCollagen, the most abundant protein in the animal kingdom, represents a promising biomaterial for regenerative medicine applications due to its structural diversity and self-assembling complexity. Despite collagen’s widely known structural and functional features, the thermodynamics behind its fibrillogenic self-assembling process is still to be fully understood. In this work we report on a series of spectroscopic, mechanical, morphological and thermodynamic characterizations of high purity type I collagen (with a D-pattern of 65 nm) extracted from Wistar Hannover rat tail. Our herein reported results can be of help to elucidate differences in self-assembly states of proteins using ITC to improve the design of energy responsive and dynamic materials for applications in tissue engineering and regenerative medicine.MethodsHerein we report the systematic study on the self-assembling fibrillogenesis mechanism of type I collagen, we provide morphological and thermodynamic evidence associated to different self-assembly events using ITC titrations. We provide thorough characterization of the effect of pH, effect of salts and protein conformation on self-assembled collagen samples via several complementary biophysical techniques, including circular dichroism (CD), Fourier Transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), atomic force microscopy (AFM), scanning electron microscopy (SEM), dynamic mechanical thermal analysis (DMTA) and thermogravimetric analysis (TGA).ResultsEmphasis was made on the use of isothermal titration calorimetry (ITC) for the thermodynamic monitoring of fibrillogenesis stages of the protein. An overall self-assembly enthalpy value of 3.27 ± 0.85 J/mol was found. Different stages of the self-assembly mechanism were identified, initial stages take place at pH values lower than the protein isoelectric point (pI), however, higher energy release events were recorded at collagen's pI. Denatured collagen employed as a control exhibited higher energy absorption at its pI, suggesting different energy exchange mechanisms as a consequence of different aggregation routes.

scholarly journals Self-Assembly Study of Type I Collagen Extracted from Male Wistar Hannover Rat Tail Tendons

2020 ◽  
Author(s):  
Jeimmy C. González-Masís ◽  
Jorge M. Cubero-Sesin ◽  
Yendry Regina Corrales-Ureña ◽  
Simón Guerrero ◽  
Sara González-Camacho ◽  
...  
Keyword(s):  
Self Assembly ◽  
Type I Collagen ◽  
Collagen Type I ◽  
The Body ◽  
Titration Calorimetry ◽  
Type I ◽  
Structural Protein ◽  
Calorimetric Analysis ◽  
Main Component ◽  
Rat Tail

Abstract Background: Collagen is the main structural protein in the extracellular matrix in the numerous connective tissues in the body. As the main component of connective tissue, it is the most abundant protein in mammals. Method: Collagen Type I from Wistar Hannover rat tail was obtained and it allowed to describe the thermodynamic profile during self-assembly by isothermal titration calorimetry (Nano ITC) as a novel technique. Results: The enthalpy of self-assembly of the 0,01035 moles of collagen was 33,89 mJ; relevant data in tissue engineering and 3D collagen fibrils bio-impressions organs, with a periodicity of 65 nm were obtained and characterized. Conclusions: Calorimetric analysis shows higher energy release event at the isoelectric point of the protein, suggesting an overall exothermic process due to self-assembly of the collagen and an endothermic process due to aggregation of denatured collagen.

Surface Modification of Polydimethylsiloxane Using Low Pressure Chemical Vapour Deposition of Poly-Chloro-p-Xylene

2012 ◽  
Vol 20 ◽  
pp. 129-142 ◽  
Author(s):  
Paul Emile Poleni ◽  
Nazare Pereira-Rodrigues ◽  
Denis Guimard ◽  
Yasuhiko Arakawa ◽  
Yasuyuki Sakai ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Self Assembly ◽  
Vapour Deposition ◽  
Type I Collagen ◽  
Chemical Vapour ◽  
The Self ◽  
Type I ◽  
Liver Tissue Engineering ◽  
Pressure Chemical ◽  
Huvec Cells

The capability to understand and modulate accurately the self-assembly of the extracellular matrix (ECM) components still one of the major fundamental objectives in the field of liver tissue engineering. In the present study, we put in evidence the suitability of poly-chloro-p-xylene (Parylene-C, ParC) for modulating the self-assembly of ECM (type-I collagen) microenvironment and cellular topography of human hepatocarcinoma (HepG2) and Human umbilical vascular endothelial (HUVEC) cells while coated on a polydimethylsiloxane (PDMS) substratum. Our findings demonstrated that the wettability of PDMS and ParC/PDMS were identical, while ParC/PDMS was significantly rougher than PDMS before and after collagen coating. However, the roughness and the wettability of ParC/PDMS were comparable to those of polystyrene (PS), a substratum commonly used for in vitro biological-related investigations. Type-I collagen adsorbed on ParC/PDMS and PS exhibited a dense network of microstructures around ~1 nm high and ~30-50 nm wide, whereas collagen adsorbed on PDMS had a low surface density of elongated fibrils that were ~2 nm thick and ~200 nm wide. This disparity in ECM microarchitecture leaded to distinct culture topographies of HepG2 cells (3D and 2D for PDMS and ParC/PDMS, respectively) and viability of HUVEC (2D viable HUVEC cells and non attached dead cells on ParC/PDMS and PDMS, respectively). To conclude, the observed changes in cell morphology and viability between ParC/PDMS and PDMS alone were directly related to the nature of the material which may impact the supramolecular organization of adsorbed ECM. We strongly believe that Low Pressure Chemical Vapour deposition (LPCVD) of ParC will offer promising insights into how microscale ECM modifications directly impact cell morphology and activity, leading to the development of advanced micro/nanosized tissue-engineered ParC/PDMS patterns with applications for liver tissue engineering.

scholarly journals A thermoreversible, photocrosslinkable collagen bio-ink for free-form fabrication of scaffolds for regenerative medicine

TECHNOLOGY ◽  
2017 ◽  
Vol 05 (04) ◽  
pp. 185-195 ◽  
Author(s):  
Kathryn E. Drzewiecki ◽  
Juilee N. Malavade ◽  
Ijaz Ahmed ◽  
Christopher J. Lowe ◽  
David I. Shreiber
Keyword(s):  
Tissue Engineering ◽  
Regenerative Medicine ◽  
Self Assembly ◽  
Type I Collagen ◽  
The Body ◽  
Free Form ◽  
Type I ◽  
Freeze Dried ◽  
Good Pattern ◽  
Further Development

As a biomaterial, collagen has been used throughout tissue engineering and regenerative medicine. Collagen is native to the body, is highly biocompatible, and naturally promotes cell adhesion and regeneration. However, collagen fibers and the inherent weak mechanical properties of collagen hydrogels interfere with further development of collagen as a bio-ink. Herein, we demonstrate the use of a modified type-I collagen, collagen methacrylamide (CMA), as a fibril-forming bio-ink for free-form fabrication of scaffolds. Like collagen, CMA can self-assemble into a fibrillar hydrogel at physiological conditions. In contrast, CMA is photocrosslinkable and thermoreversible, and photocrosslinking eliminates thermoreversibility. Free-form fabrication of CMA was performed through self-assembly of the CMA hydrogel, photocrosslinking the structure of interest using a photomask, and cooling the entire hydrogel, which results in cold-melting of unphotocrosslinked regions. Printed hydrogels had a resolution on the order of [Formula: see text]350[Formula: see text][Formula: see text]m, and can be fabricated with or without cells and maintain viability or be further processed into freeze-dried sponges, all while retaining pattern fidelity. A subcutaneous implant study confirmed the biocompatibility of CMA in comparison to collagen. Free-form fabrication of CMA allows for printing of macroscale, customized scaffolds with good pattern fidelity and can be implemented with relative ease for continued research and development of collagen-based scaffolds in tissue engineering.

scholarly journals Marangoni flows drive the alignment of fibrillar cell-laden hydrogels

2020 ◽  
Vol 6 (24) ◽  
pp. eaaz7748 ◽  
Author(s):  
Bryan A. Nerger ◽  
P.-T. Brun ◽  
Celeste M. Nelson
Keyword(s):  
Self Assembly ◽  
Collagen Fiber ◽  
Type I Collagen ◽  
Type I ◽  
Sessile Droplet ◽  
Biomimetic Materials ◽  
Fiber Assembly ◽  
Self Assembling ◽  
Surrounding Environment ◽  
Marangoni Flows

When a sessile droplet containing a solute in a volatile solvent evaporates, flow in the droplet can transport and assemble solute particles into complex patterns. Transport in evaporating sessile droplets has largely been examined in solvents that undergo complete evaporation. Here, we demonstrate that flow in evaporating aqueous sessile droplets containing type I collagen—a self-assembling polymer—can be harnessed to engineer hydrated networks of aligned collagen fibers. We find that Marangoni flows direct collagen fiber assembly over millimeter-scale areas in a manner that depends on the rate of self-assembly, the relative humidity of the surrounding environment, and the geometry of the droplet. Skeletal muscle cells that are incorporated into and cultured within these evaporating droplets collectively orient and subsequently differentiate into myotubes in response to aligned networks of collagen. Our findings demonstrate a simple, tunable, and high-throughput approach to engineer aligned fibrillar hydrogels and cell-laden biomimetic materials.

Directive Effect of Chain Length in Modulating Peptide Nano-assemblies

2020 ◽  
Vol 27 (9) ◽  
pp. 923-929
Author(s):  
Gaurav Pandey ◽  
Prem Prakash Das ◽  
Vibin Ramakrishnan
Keyword(s):  
Electron Microscopy ◽  
Amino Acids ◽  
Light Scattering ◽  
Chain Length ◽  
Self Assembly ◽  
The Self ◽  
Ionic Charge ◽  
Self Assembling ◽  
Biophysical Techniques

Background: RADA-4 (Ac-RADARADARADARADA-NH2) is the most extensively studied and marketed self-assembling peptide, forming hydrogel, used to create defined threedimensional microenvironments for cell culture applications. Objectives: In this work, we use various biophysical techniques to investigate the length dependency of RADA aggregation and assembly. Methods: We synthesized a series of RADA-N peptides, N ranging from 1 to 4, resulting in four peptides having 4, 8, 12, and 16 amino acids in their sequence. Through a combination of various biophysical methods including thioflavin T fluorescence assay, static right angle light scattering assay, Dynamic Light Scattering (DLS), electron microscopy, CD, and IR spectroscopy, we have examined the role of chain-length on the self-assembly of RADA peptide. Results: Our observations show that the aggregation of ionic, charge-complementary RADA motifcontaining peptides is length-dependent, with N less than 3 are not forming spontaneous selfassemblies. Conclusion: The six biophysical experiments discussed in this paper validate the significance of chain-length on the epitaxial growth of RADA peptide self-assembly.

Self-assembling behaviour of a modified aromatic amino acid in competitive medium

Soft Matter ◽  
2020 ◽  
Vol 16 (28) ◽  
pp. 6599-6607 ◽  
Author(s):  
Pijush Singh ◽  
Souvik Misra ◽  
Nayim Sepay ◽  
Sanjoy Mondal ◽  
Debes Ray ◽  
...  
Keyword(s):  
Amino Acid ◽  
Self Assembly ◽  
Aromatic Amino Acid ◽  
Photophysical Properties ◽  
Solvent Mixture ◽  
The Self ◽  
Solvent Ratio ◽  
Self Assembling

The self-assembly and photophysical properties of 4-nitrophenylalanine (4NP) are changed with the alteration of solvent and final self-assembly state of 4NP in competitive solvent mixture and are dictated by the solvent ratio.

Sign in / Sign up

Export Citation Format

Share Document