scholarly journals Increased Fibroblast Metabolic Activity of Collagen Scaffolds via the Addition of Propolis Nanoparticles

Materials ◽  
2020 ◽  
Vol 13 (14) ◽  
pp. 3118
Author(s):  
Jeimmy González-Masís ◽  
Jorge M. Cubero-Sesin ◽  
Yendry R. Corrales-Ureña ◽  
Sara González-Camacho ◽  
Nohelia Mora-Ugalde ◽  
...  
Keyword(s):  
Metabolic Activity ◽  
Self Assembly ◽  
Type I Collagen ◽  
Fibroblast Cell ◽  
Antimicrobial Activities ◽  
Hydrodynamic Diameter ◽  
Type I ◽  
Collagen Scaffolds ◽  
Simple Method ◽  
Socially Relevant

Propolis natural extracts have been used since ancient times due to their antioxidant, anti-inflammatory, antiviral, and antimicrobial activities. In this study, we produced scaffolds of type I collagen, extracted from Wistar Hanover rat tail tendons, and impregnated them with propolis nanoparticles (NPs) for applications in regenerative medicine. Our results show that the impregnation of propolis NPs to collagen scaffolds affected the collagen denaturation temperature and tensile strength. The changes in structural collagen self-assembly due to contact with organic nanoparticles were shown for the first time. The fibril collagen secondary structure was preserved, and the D-pattern gap increased to 135 ± 28 nm, without losing the microfiber structure. We also show that the properties of the collagen scaffolds depended on the concentration of propolis NPs. A concentration of 100 μg/mL of propolis NPs with 1 mg of collagen, with a hydrodynamic diameter of 173 nm, was found to be an optimal concentration to enhance 3T3 fibroblast cell metabolic activity and cell proliferation. The expected outcome from this research is both scientifically and socially relevant since the home scaffold using natural nanoparticles can be produced using a simple method and could be widely used for local medical care in developing communities.

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 Experimental and Modeling Study of Collagen Scaffolds with the Effects of Crosslinking and Fiber Alignment

2011 ◽  
Vol 2011 ◽  
pp. 1-12 ◽  
Author(s):  
Bin Xu ◽  
Ming-Jay Chow ◽  
Yanhang Zhang
Keyword(s):  
Magnetic Particles ◽  
Type I Collagen ◽  
Collagen Gel ◽  
Collagen Type I ◽  
Mechanical Tests ◽  
Type I ◽  
Collagen Scaffolds ◽  
Fiber Alignment ◽  
Biaxial Tensile ◽  
Anisotropic Mechanical Properties

Collagen type I scaffolds are commonly used due to its abundance, biocompatibility, and ubiquity. Most applications require the scaffolds to operate under mechanical stresses. Therefore understanding and being able to control the structural-functional integrity of collagen scaffolds becomes crucial. Using a combined experimental and modeling approach, we studied the structure and function of Type I collagen gel with the effects of spatial fiber alignment and crosslinking. Aligned collagen scaffolds were created through the flow of magnetic particles enmeshed in collagen fibrils to mimic the anisotropy seen in native tissue. Inter- and intra- molecular crosslinking was modified chemically with Genipin to further improve the stiffness of collagen scaffolds. The anisotropic mechanical properties of collagen scaffolds were characterized using a planar biaxial tensile tester and parallel plate rheometer. The tangent stiffness from biaxial tensile test is two to three orders of magnitude higher than the storage moduli from rheological measurements. The biphasic nature of collagen gel was discussed and used to explain the mechanical behavior of collagen scaffolds under different types of mechanical tests. An anisotropic hyperelastic constitutive model was used to capture the characteristics of the stress-strain behavior exhibited by collagen scaffolds.

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.

Modifying the Properties of Collagen Scaffolds with Microfluidics

2005 ◽  
Vol 897 ◽  
Author(s):  
David I Shreiber ◽  
Harini G Sundararaghavan ◽  
Minjung Song ◽  
Vikram Munikoti ◽  
Kathryn E Uhrich
Keyword(s):  
Mechanical Properties ◽  
Cell Adhesion ◽  
Cellular Response ◽  
Type I Collagen ◽  
Crosslinking Agent ◽  
Force Balance ◽  
Adherent Cell ◽  
Type I ◽  
Spinal Cord Regeneration ◽  
Collagen Scaffolds

AbstractIt is now well accepted that the mechanical properties and cell adhesion profile of 2D and 3D extracellular matrix molecules combine to dictate cellular fate processes, such as differentiation, migration, proliferation, and apoptosis, through a process generally known as 'mechanotransduction', or the conversion of mechanical signals into a cellular response. The stiffness and adhesion density combine to affect the force balance that exists between an adherent cell and the surrounding substrate. We have established BioMEMS, microfluidic technology to alter the mechanical properties and cell adhesion profile of collagen scaffolds. Using soft lithography, we fabricate elastomeric networks that serve as conduits for the controlled mixing of type I collagen solutions. Our technology enables us to generate reproducible, controlled homogeneous and inhomogeneous microenvironments for 3D cell culture, assays of cell behavior in 3D, and the development of bioartificial tissue equivalents for regenerative and reparative therapies. The adhesivity of collagen is modulated by covalently grafting peptides (such as RGD) or proteins (such as albumin) to soluble collagen molecules with 1- ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC), a hetero-bifunctional coupling agent. EDC activates the carboxylic group of collagen and forms an amine bond with the grafting molecule. The grafted collagen self-assembles into a fibrillar gel at physiological temperature and pH with no measurable changes in rheological properties compared to controls. A solution of peptide-grafted collagen is then mixed in microfluidic networks with unaltered collagen to form controlled gradients or other patterns of the two solutions, which immobilize upon self-assembly. Separately or in the same network, the mechanical properties of the collagen gel can be altered regionally by the microfluidic delivery a solution of a cell-tolerated crosslinking agent. We use genipin, which has the unique property of generating crosslinks that autofluoresce. The intensity of the fluorescence correlates with the degree of crosslinking (and thus the mechanical properties) enabling us to monitor and measure changes in mechanical properties dynamically and non-invasively. Lastly, though it requires constant delivery or recirculation, the same networks can be used to impose gradients of soluble factors, such as growth factors and cytokines. Thus, we have developed a platform to examine the response of cells to simultaneous chemotactic, haptotactic, and durotactic gradients in a 3D environment. We are employing this technology to examine the response of neural cells to gradients of biomaterial properties to optimize cues for spinal cord regeneration.

Mechanics and Cytocompatibility of Genipin Crosslinked Type I Collagen Nanofibrous Scaffolds

2008 ◽  
Author(s):  
Albert O. Gee ◽  
Brendon M. Baker ◽  
Robert L. Mauck
Keyword(s):  
Type I Collagen ◽  
Cell Attachment ◽  
Type I ◽  
Primary Role ◽  
Fiber Reinforced ◽  
Collagen Scaffolds ◽  
Major Drawback ◽  
Aligned Arrays ◽  
Musculoskeletal Tissues ◽  
Nanofibrous Scaffolds

Collagen is a principal constituent of the extracellular matrix (ECM) and as such, defines the microenvironmental milieu in which cells reside. In fiber-reinforced musculoskeletal tissues, collagen fibers are highly organized and generate the direction-dependent mechanical properties critical to the function of these structures. Given its primary role, collagen is particularly attractive for tissue engineering (TE) applications where scaffolds are coupled with cells to repair or regenerate damaged tissues. One method for producing collagen-based scaffolds is through electrospinning. This technique yields nano- to micron-scale fibers similar in diameter to those of the native ECM. Towards engineering orthopaedic tissues, methods have been devised to electrospin fibers into aligned arrays that can recapitulate the anisotropy of fiber-reinforced tissues [1]. While a number of polymers have been electrospun, collagen-based scaffolds are especially promising as they provide a biomimetic interface for cell attachment [2]. Numerous investigators have electrospun collagen [3], one major drawback is their inherent instability in aqueous environments. To address this, various crosslinking agents including glutaraldehyde (GA), 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride, and N-hydroxysuccinimide chemistries have been used, but these chemicals often prove cytotoxic or excessively laborious in application [4]. Even with crosslinking, dry as-formed nanofibrous collagen scaffolds with moduli greater than 50MPa diminish by 100-fold with rehydration [5].

scholarly journals Treatment with type-I collagen scaffolds in patients with venous ulcers. Case report

Case reports ◽  
2020 ◽  
Vol 6 (2) ◽  
pp. 128-136
Author(s):  
Martha Isabel González-Duque ◽  
Julián Daniel Hernández-Martínez ◽  
Marta Raquel Fontanilla ◽  
Sofía Elizabeth Muñoz-Medina
Keyword(s):  
Type I Collagen ◽  
Low Cost ◽  
Adult Population ◽  
Lower Limbs ◽  
Collagen Membrane ◽  
Type I ◽  
Dermal Substitute ◽  
Collagen Scaffolds ◽  
Venous Ulcers

Introduction: Chronic venous insufficiency affects about 5% of the global adult population. Venous leg ulcers are one of the most frequent complications of this pathology, with a global prevalence of 2%. This disease affects both the quality of life of patients and, due to the high cost of the treatment, the health system. Compressive therapy and moist wound healing have been the gold standard treatment. However, when complications occur, they may not be effective.Case report: This is the case of a 66-year-old female patient with venous ulcers on her lower limbs and symptoms of fever and local pain that did not respond to conventional therapies. The patient was treated with a new dermal substitute made of an acellular type-I collagen membrane, which promotes the closure of the ulcer by stimulating the replacement of injured tissue with tissue similar to the healthy one. The condition of the patient improved at 16 weeks, and after 8 months of treatment there was no recurrence of the lesions.Conclusions: Acellular type-I collagen membrane developed by the Tissue Engineering Working Group of the Department of Pharmacy of the Universidad Nacional de Colombia is effective in treating venous ulcers of the lower limbs. Its low cost facilitates the access of the whole population to therapies based on its application.

Influence of Mechanical Load on the Degradation of Corneal Collagen

2008 ◽  
Author(s):  
Ramin Zareian ◽  
Kelli P. Church ◽  
Jeffrey W. Ruberti
Keyword(s):  
Self Assembly ◽  
Type I Collagen ◽  
Mechanical Load ◽  
Structural Performance ◽  
Type I ◽  
Connective Tissues ◽  
Bacterial Collagenase ◽  
Collagen Triple Helix ◽  
Different Types ◽  
Corneal Collagen

Collagen is one of the most important structural proteins in vertebrate animals. Over 25 different types of collagen have been identified, but type I collagen is the most abundant fibril forming collagen and contributes to the structural performance numerous connective tissues including ligaments, tendons and cornea [1]. In addition to collagen self-assembly, collagen degradation is an important step in the development, remodeling, homeostasis and pathology of load-bearing ECM. Matrix Metalloproteinase (MMP) types I and VIII, bacterial collagenase and cathepsin are the best known enzymes capable of directly degrading the collagen triple helix [2, 3]. Several researchers have hypothesized that straining collagen fibrils makes them less susceptible to enzymatic degradation [4, 5]. This concept, which we refer to as “strain-stabilization” has important implications for our understanding of collagen as an engineering material.

scholarly journals The Characteristics of Intrinsic Fluorescence of Type I Collagen Influenced by Collagenase I

2018 ◽  
Vol 8 (10) ◽  
pp. 1947 ◽  
Author(s):  
Yiming Shen ◽  
Deyi Zhu ◽  
Wenhui Lu ◽  
Bing Liu ◽  
Yanchun Li ◽  
...  
Keyword(s):  
Fluorescence Intensity ◽  
Self Assembly ◽  
Triple Helix ◽  
Type I Collagen ◽  
Intrinsic Fluorescence ◽  
Type I ◽  
Food And Agriculture ◽  
Mechanism Research ◽  
Helix Structure ◽  
Assembly Behavior

The triple helix structure of collagen can be degraded by collagenase. In this study, we explored how the intrinsic fluorescence of type I collagen was influenced by collagenase I. We found that tyrosine was the main factor that could successfully excite the collagen fluorescence. Initially, self-assembly behavior of collagen resulted in a large amount of tyrosine wrapped with collagen, which decreased the fluorescence intensity of type I collagen. After collagenase cleavage, some wrapped-tyrosine could be exposed and thereby the intrinsic fluorescence intensity of collagen increased. By observation and analysis, the influence of collagenase to intrinsic fluorescence of collagen was investigated and elaborated. Furthermore, collagenase cleavage to the special triple helix structure of collagen would result in a slight improvement of collagen thermostability, which was explained by the increasing amount of terminal peptides. These results are helpful and effective for reaction mechanism research related to collagen, which can be observed by fluorescent technology. Meantime, the reaction behaviors of both collagenase and collagenolytic proteases can also be analyzed by fluorescent technology. In conclusion, this research provides a foundation for the further investigation of collagen reactions in different areas, such as medicine, nutrition, food and agriculture.

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