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 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

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 The Effect of Collagen-I Coatings of 3D Printed PCL Scaffolds for Bone Replacement on Three Different Cell Types

2021 ◽  
Vol 11 (22) ◽  
pp. 11063
Author(s):  
Lucas Weingärtner ◽  
Sergio H. Latorre ◽  
Dirk Velten ◽  
Anke Bernstein ◽  
Hagen Schmal ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Type I Collagen ◽  
Testing Machine ◽  
Cell Types ◽  
The Body ◽  
Type I ◽  
Human Tissues ◽  
Cytotoxicity Studies ◽  
Cell Scaffold ◽  
Different Cell Types

Introduction The use of scaffolds in tissue engineering is becoming increasingly important as solutions need to be found to preserve human tissues such as bone or cartilage. Various factors, including cells, biomaterials, cell and tissue culture conditions, play a crucial role in tissue engineering. The in vivo environment of the cells exerts complex stimuli on the cells, thereby directly influencing cell behavior, including proliferation and differentiation. Therefore, to create suitable replacement or regeneration procedures for human tissues, the conditions of the cells’ natural environment should be well mimicked. Therefore, current research is trying to develop 3-dimensional scaffolds (scaffolds) that can elicit appropriate cellular responses and thus help the body regenerate or replace tissues. In this work, scaffolds were printed from the biomaterial polycaprolactone (PCL) on a 3D bioplotter. Biocompatibility testing was used to determine whether the printed scaffolds were suitable for use in tissue engineering. Material and Methods An Envisiontec 3D bioplotter was used to fabricate the scaffolds. For better cell-scaffold interaction, the printed polycaprolactone scaffolds were coated with type-I collagen. Three different cell types were then cultured on the scaffolds and various tests were used to investigate the biocompatibility of the scaffolds. Results Reproducible scaffolds could be printed from polycaprolactone. In addition, a coating process with collagen was developed, which significantly improved the cell-scaffold interaction. Biocompatibility tests showed that the PCL-collagen scaffolds are suitable for use with cells. The cells adhered to the surface of the scaffolds and as a result extensive cell growth was observed on the scaffolds. The inner part of the scaffolds, however, remained largely uninhabited. In the cytotoxicity studies, it was found that toxicity below 20% was present in some experimental runs. The determination of the compressive strength by means of the universal testing machine Z005 by ZWICK according to DIN EN ISO 604 of the scaffolds resulted in a value of 68.49 ± 0.47 MPa.

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.

scholarly journals Collagen-Based Electrospun Materials for Tissue Engineering: A Systematic Review

2021 ◽  
Vol 8 (3) ◽  
pp. 39
Author(s):  
Britani N. Blackstone ◽  
Summer C. Gallentine ◽  
Heather M. Powell
Keyword(s):  
Systematic Review ◽  
Tissue Engineering ◽  
Collagen Type I ◽  
The Body ◽  
Pool Data ◽  
Type I ◽  
High Concentrations ◽  
Increased Strength

Collagen is a key component of the extracellular matrix (ECM) in organs and tissues throughout the body and is used for many tissue engineering applications. Electrospinning of collagen can produce scaffolds in a wide variety of shapes, fiber diameters and porosities to match that of the native ECM. This systematic review aims to pool data from available manuscripts on electrospun collagen and tissue engineering to provide insight into the connection between source material, solvent, crosslinking method and functional outcomes. D-banding was most often observed in electrospun collagen formed using collagen type I isolated from calfskin, often isolated within the laboratory, with short solution solubilization times. All physical and chemical methods of crosslinking utilized imparted resistance to degradation and increased strength. Cytotoxicity was observed at high concentrations of crosslinking agents and when abbreviated rinsing protocols were utilized. Collagen and collagen-based scaffolds were capable of forming engineered tissues in vitro and in vivo with high similarity to the native structures.

Tissue Engineering of Bone by Osteoinductive Biomaterials

MRS Bulletin ◽  
1996 ◽  
Vol 21 (11) ◽  
pp. 36-39 ◽  
Author(s):  
Ugo Ripamonti ◽  
Nicolaas Duneas
Keyword(s):  
Tissue Engineering ◽  
Bone Formation ◽  
Type I Collagen ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Mesenchymal Cells ◽  
Tissue Response ◽  
Type I ◽  
New Bone Formation ◽  
Demineralized Bone

Recent advances in materials science and biotechnology have given birth to the new and exciting field of tissue engineering, in which the two normally disparate fields are merging into a profitable matrimony. In particular the use of biomaterials capable of initiating new bone formation via a process called osteoinduction is leading to quantum leaps for the tissue engineering of bone.The classic work of Marshall R. Urist and A. Hari Reddi opened the field of osteoinductive biomaterials. Urist discovered that, upon implantation of devitalized, demineralized bone matrix in the muscle of experimental animals, new bone formation occurs within two weeks, a phenomenon he described as bone formation by induction. The tissue response elicited by implantation of demineralized bone matrix in muscle or under the skin includes activation and migration of undifferentiated mesenchymal cells by chemotaxis, anchoragedependent cell attachment to the matrix, mitosis and proliferation of mesenchymal cells, differentiation of cartilage, mineralization of the cartilage, vascular invasion of the cartilage, differentiation of osteoblasts and deposition of bone matrix, and finally mineralization of bone and differentiation of marrow in the newly developed ossicle.The osteoinductive ability of the extracellular matrix of bone is abolished by the dissociative extraction of the demineralized matrix, but is recovered when the extracted component, itself inactive, is reconstituted with the inactive residue—mainly insoluble collagenous bone matrix. This important experiment showed that the osteoinductive signal resides in the solubilized component but needs to be reconstituted with an appropriate carrier to restore the osteoinductive activity. In this case, the carrier is the insoluble collagenous bone matrix—mainly crosslinked type I collagen.

Macromolecular Crowding Meets Tissue Engineering by Self-Assembly: A Paradigm Shift in Regenerative Medicine

2014 ◽  
Vol 26 (19) ◽  
pp. 3024-3034 ◽  
Author(s):  
Abhigyan Satyam ◽  
Pramod Kumar ◽  
Xingliang Fan ◽  
Alexander Gorelov ◽  
Yury Rochev ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Regenerative Medicine ◽  
Self Assembly ◽  
Paradigm Shift ◽  
Macromolecular Crowding

Tissue engineering of dental pulp on type I collagen

2004 ◽  
Vol 29 (4) ◽  
pp. 370
Author(s):  
Gwang-Hee Lee ◽  
Sung-Yoon Huh ◽  
Sang-Hyuk Park
Keyword(s):  
Tissue Engineering ◽  
Dental Pulp ◽  
Type I Collagen ◽  
Type I

Regulation of collagen I gene expression by ras

1992 ◽  
Vol 12 (10) ◽  
pp. 4714-4723
Author(s):  
J L Slack ◽  
M I Parker ◽  
V R Robinson ◽  
P Bornstein
Keyword(s):  
Type I Collagen ◽  
The Body ◽  
Transformed Cells ◽  
Type I ◽  
Mrna Levels ◽  
Oncogenic Ras ◽  
Genes Encoding ◽  
Flanking Region ◽  
Collagen Genes ◽  
I Gene

Although transformation of rodent fibroblasts can lead to dramatic changes in expression of extracellular matrix genes, the molecular basis and physiological significance of these changes remain poorly understood. In this study, we have investigated the mechanism(s) by which ras affects expression of the genes encoding type I collagen. Levels of both alpha 1(I) and alpha 2(I) collagen mRNAs were markedly reduced in Rat 1 fibroblasts overexpressing either the N-rasLys-61 or the Ha-rasVal-12 oncogene. In fibroblasts conditionally transformed with N-rasLys-61, alpha 1(I) transcript levels began to decline within 8 h of ras induction and reached 1 to 5% of control levels after 96 h. In contrast, overexpression of normal ras p21 had no effect on alpha 1(I) or alpha 2(I) mRNA levels. Nuclear run-on experiments demonstrated that the transcription rates of both the alpha 1(I) and alpha 2(I) genes were significantly reduced in ras-transformed cells compared with those in parental cells. In addition, the alpha 1(I) transcript was less stable in transformed cells. Chimeric plasmids containing up to 3.6 kb of alpha 1(I) 5'-flanking DNA and up to 2.3 kb of the 3'-flanking region were expressed at equivalent levels in both normal and ras-transformed fibroblasts. However, a cosmid clone containing the entire mouse alpha 1(I) gene, including 3.7 kb of 5'- and 4 kb of 3'-flanking DNA, was expressed at reduced levels in fibroblasts overexpressing oncogenic ras. We conclude that oncogenic ras regulates the type I collagen genes at both transcriptional and posttranscriptional levels and that this effect, at least for the alpha 1(I) gene, may be mediated by sequences located either within the body of the gene itself or in the distal 3'-flanking region.

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