The Effect of Crosslinking Agents on the Properties of Type II Collagen Biomaterials

2021 ◽  
Vol 57 (4) ◽  
pp. 166-180
Author(s):  
Maria-Minodora Marin ◽  
Madalina Georgiana Albu Kaya ◽  
George Mihail Vlasceanu ◽  
Jana Ghitman ◽  
Ionut Cristian Radu ◽  
...  

Type II collagen has been perceived as the indispensable element and plays a crucial role in cartilage tissue engineering. Thus, materials based on type II collagen have drawn farther attention in both academic and research for developing new systems for the cartilage regeneration. The disadvantage of using type II collagen as a biomaterial for tissue repairing is its reduced biomechanical properties. This can be solved by physical, enzymatic or chemical cross-linking processes, which provide biomaterials with the required mechanical properties for medical applications. To enhance type II collagen properties, crosslinked collagen scaffolds with different cross-linking agents were prepared by freeze-drying technique. The present research work studied the synthesis of type II collagen biomaterials with and without crosslinking agents. Scaffolds morphology was observed by MicroCT, showing in all cases an appropriate microstructure for biological applications, and the mechanical studies were performed using compressive tests. DSC showed an increase in denaturation temperature with an increase in cross-linking agent concentration. FTIR suggested that the secondary structure of collagen is not affected after the cross-linking; supplementary, to confirm the characteristic triple-helix conformation of collagen, the CD investigation was performed. The results showed that the physical-chemical properties of type II collagen were improved by cross-linking treatments.

Polymers ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 4199
Author(s):  
Mahshid Hafezi ◽  
Saied Nouri Khorasani ◽  
Mohadeseh Zare ◽  
Rasoul Esmaeely Neisiany ◽  
Pooya Davoodi

Cartilage is a tension- and load-bearing tissue and has a limited capacity for intrinsic self-healing. While microfracture and arthroplasty are the conventional methods for cartilage repair, these methods are unable to completely heal the damaged tissue. The need to overcome the restrictions of these therapies for cartilage regeneration has expanded the field of cartilage tissue engineering (CTE), in which novel engineering and biological approaches are introduced to accelerate the development of new biomimetic cartilage to replace the injured tissue. Until now, a wide range of hydrogels and cell sources have been employed for CTE to either recapitulate microenvironmental cues during a new tissue growth or to compel the recovery of cartilaginous structures via manipulating biochemical and biomechanical properties of the original tissue. Towards modifying current cartilage treatments, advanced hydrogels have been designed and synthesized in recent years to improve network crosslinking and self-recovery of implanted scaffolds after damage in vivo. This review focused on the recent advances in CTE, especially self-healing hydrogels. The article firstly presents the cartilage tissue, its defects, and treatments. Subsequently, introduces CTE and summarizes the polymeric hydrogels and their advances. Furthermore, characterizations, the advantages, and disadvantages of advanced hydrogels such as multi-materials, IPNs, nanomaterials, and supramolecular are discussed. Afterward, the self-healing hydrogels in CTE, mechanisms, and the physical and chemical methods for the synthesis of such hydrogels for improving the reformation of CTE are introduced. The article then briefly describes the fabrication methods in CTE. Finally, this review presents a conclusion of prevalent challenges and future outlooks for self-healing hydrogels in CTE applications.


2017 ◽  
Vol 23 (1-2) ◽  
pp. 55-68 ◽  
Author(s):  
Henrique V. Almeida ◽  
Binulal N. Sathy ◽  
Ivan Dudurych ◽  
Conor T. Buckley ◽  
Fergal J. O'Brien ◽  
...  

2007 ◽  
Vol 342-343 ◽  
pp. 89-92 ◽  
Author(s):  
Jae Ho Jeong ◽  
Y.M. Moon ◽  
S.O. Kim ◽  
S.S. Yun ◽  
Hong In Shin

Despite many outstanding research works on cartilage tissue engineering, actual clinical application is not quite successful because of the absorption and progressive distortion of tissue engineered cartilage. We have developed a new method of cartilage tissue engineering comprising chondrocyte mixed Pluronic F-127 and cultured chondrocyte cell sheet which entirely cover the cell-Pluronic complex. We believe the addition of cultured chondrocyte cell sheet enhances the efficacy of chondrogenesis in vivo. Human ear cartilage piece was enzymatically dissociated and chondrocyte suspension was acquired. Chondrocytes were cultured and expanded as the routine manner. Cultured chondrocytes were plated in high-density monolayer and cultured with Chondrogenic media in 5% CO2 incubator. After 3 weeks of culture, chondrocyte cell sheet was formed and complete single sheet of chondrocyte could be harvested by gentle manipulation of culture plate with a cell scraper. Chondrocyte-Pluronic complex was established by mixing 1x 106 cells with 0.5 of Pluronic F- 127. Chondrocyte-Pluronic complex was completely covered with a sheet of cultured chondrocyte. The completed tissue engineered constructs were implanted into the subcutaneous tissue pocket of nude mice on the back. Tissue engineered constructs without cultured cell sheet were used as control. Samples were harvested at 8 weeks postoperatively and they were subjected to histological analysis and assayed for glycosaminoglycan (GAG), and type II collagen. Grossly, the size of cartilage specimen of cultured chondrocyte cell sheet covered group was larger than that of the control. On histologic examination, the specimen of cultured chondrocyte cell sheet covered group showed lacunae-containing cells embedded in a basophilic matrix. The chondrocyte cell sheet covered group specimen resembled mature or immature cartilage. The result of measurement of GAG and type II collagen of cartilage specimen of cultured chondrocyte sheet covered group was higher than that of the control. In conclusion, the new method of cartilage tissue engineering using chondrocyte cell sheet seems to be an effective method providing higher cartilage tissue gain and reliable success rate for cartilage tissue engineering.


2008 ◽  
Vol 31 (11) ◽  
pp. 960-969 ◽  
Author(s):  
H.J. Pulkkinen ◽  
V. Tiitu ◽  
P. Valonen ◽  
E.-R. Hämäläinen ◽  
M.J. Lammi ◽  
...  

Purpose Collagen type II is the major component of cartilage and would be an optimal scaffold material for reconstruction of injured cartilage tissue. In this study, the feasibility of recombinant human type II collagen gel as a 3-dimensional culture system for bovine chondrocytes was evaluated in vitro. Methods Bovine chondrocytes (4x106 cells) were seeded within collagen gels and cultivated for up to 4 weeks. The gels were investigated with confocal microscopy, histology, and biochemical assays. Results Confocal microscopy revealed that the cells maintained their viability during the entire cultivation period. The chondrocytes were evenly distributed inside the gels, and the number of cells and the amount of the extracellular matrix increased during cultivation. The chondrocytes maintained their round phenotype during the 4-week cultivation period. The glycosaminoglycan levels of the tissue increased during the experiment. The relative levels of aggrecan and type II collagen mRNA measured with realtime polymerase chain reaction (PCR) showed an increase at 1 week. Conclusion Our results imply that recombinant human type II collagen is a promising biomaterial for cartilage tissue engineering, allowing homogeneous distribution in the gel and biosynthesis of extracellular matrix components.


2011 ◽  
Vol 11 (02) ◽  
pp. 433-443 ◽  
Author(s):  
BENJAMIN D. ELDER ◽  
ARVIND MOHAN ◽  
KYRIACOS A. ATHANASIOU

Background. As articular cartilage is unable to repair itself, there is a tremendous clinical need for a tissue engineered replacement tissue. Current tissue engineering efforts using the self-assembly process have demonstrated promising results, but the biomechanical properties remain at roughly 50% of native tissue. Methodology/Principal Findings. The objective of this study was to determine the feasibility of using exogenous crosslinking agents to enhance the biomechanical properties of a scaffoldless cartilage tissue engineering approach. Four crosslinking agents (glutaraldehyde, ribose, genipin, and methylglyoxal) were applied each at a single concentration and single application time. It was determined that ribose application resulted in a significant 69% increase in Young's modulus, a significant 47% increase in ultimate tensile strength, as well as a trend toward a significant increase in aggregate modulus. Additionally, methylglyoxal application resulted in a significant 58% increase in Young's modulus. No treatments altered the biochemical content of the tissue. Conclusions/Significance. To our knowledge, this is the first study to examine the use of exogenous crosslinking agents on any tissue formed using a scaffoldless tissue engineering approach. In particular, this study demonstrates that a one-time treatment with crosslinking agents can be employed effectively to enhance the biomechanical properties of tissue engineered articular cartilage. The results are exciting, as they demonstrate the feasibility of using exogenous crosslinking agents to enhance the biomechanical properties without the need for increased glycosaminoglycan (GAG) and collagen content.


2007 ◽  
Vol 15 ◽  
pp. B136
Author(s):  
H.J. Pulkkinen ◽  
V. Tiitu ◽  
P. Valonen ◽  
E. Hämäläinen ◽  
J. Koivurinta ◽  
...  

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Dechao Yuan ◽  
Zhu Chen ◽  
Tao Lin ◽  
Xuwei Luo ◽  
Hua Dong ◽  
...  

A novel chitosan hydrogel with high porosity was fabricated by a crosslinking method. Cartilage tissue engineering formed after mesenchymal stem cells was cultured on this hydrogel scaffold for 12 weeks. The immunohistochemistry tests demonstrated that the obtained cartilage had the specific histological properties of natural cartilage. And the qPCR tests also proved that the genes for type II collagen in the obtained cartilage were expressed the same as in the natural one.


Author(s):  
Robert J. Nims ◽  
Alexander D. Cigan ◽  
Michael B. Albro ◽  
Clark T. Hung ◽  
Gerard A. Ateshian

Cartilage tissue engineering (CTE) is a strategy of great interest and promise for the replacement of osteoarthritic (OA) cartilage. In CTE, chondrocytes are used to synthesize cartilage matrix products (predominantly glycosaminoglycans (GAG) and type II collagen). The aim for CTE is to develop engineered constructs with mechanical properties and biochemical composition comparable to native tissue, to reproduce its functional properties.


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