Combined Effect of Glycosaminoglycan and Mechanical Stimulation on the In Vitro Biomechanics of Tissue Engineered Tendon Constructs

2007 ◽  
Author(s):  
Kirsten R. C. Kinneberg ◽  
Victor S. Nirmalanandhan ◽  
Heather M. Powell ◽  
Steven T. Boyce ◽  
David L. Butler
Keyword(s):  
Tissue Engineering ◽  
Type I Collagen ◽  
Rabbit Model ◽  
The United States ◽  
Maximum Force ◽  
Attractive Alternative ◽  
Type I ◽  
Post Surgery ◽  
Scaffold Materials

Tissue engineering offers an attractive alternative to direct repair or reconstruction of injuries to tendons, ligaments and capsular structures that represent almost 45% of the 32 million musculoskeletal injuries that occur each year in the United States [1]. Mesenchymal stem cell (MSC)-seeded collagen constructs are currently being used by our group to repair tendon injuries in the rabbit model [2, 3]. Although these cell-assisted repairs exhibit 50% greater maximum force and stiffness at 12 weeks compared to values for natural repair, tissues often lack the maximum force sufficient to resist the peak in vivo forces acting on the repair site [3]. Our laboratory has previously demonstrated that in vitro construct stiffness and repair stiffness at 12 weeks post surgery are positively correlated [4]. Therefore, in an effort to further improve the repair outcome using tissue engineering, we continue our investigation of scaffold materials to create stiffer MSC-collagen constructs. Our group has recently evaluated two scaffold materials, type I collagen sponges fabricated within the Engineered Skin Lab (ESL, Shriners Hospitals for Children) by a freezing and lyophilization process with and without glycosaminoglycan (chondroitin-6-sulfate; GAG) [5] and found the ESL sponges to significantly improve biomechanical properties of the constructs compared to sponges we currently use in the lab (P1076, Kensey Nash Corporation, Exton, PA). This study also demonstrated that GAG significantly upregulates collagen type I, decorin, and fibronectin gene expression (unpublished results).

Evaluation of a Novel Scaffold Material for Tendon Tissue Engineering

2007 ◽  
Author(s):  
Victor S. Nirmalanandhan ◽  
Kirsten R. C. Kinneberg ◽  
Natalia Juncosa-Melvin ◽  
Heather M. Powell ◽  
Marepalli Rao ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cell ◽  
Type I Collagen ◽  
Tendon Repair ◽  
Maximum Force ◽  
Attractive Alternative ◽  
Mechanical Resistance ◽  
Type I ◽  
Novel Scaffold

Tissue engineering offers an attractive alternative to direct repair or reconstruction of soft tissue injuries. Tissue engineered constructs containing mesenchymal stem cells (MSCs) seeded in commercially available type I collagen sponges (P1076, Kensey Nash Corporation, Exton, PA) are currently being used within our laboratory to repair tendon injuries in rabbit models [1]. When introduced into the wound site, mechanically stimulated stem cell-collagen sponge constructs exhibit 50% greater maximum force and stiffness at 12 weeks compared to values for static controls [1]. However, these constructs often lack the maximum force sufficient to resist the peak in vivo forces acting on the repair site [2, 3]. Insufficient repair biomechanics can be attributed to the poor initial mechanical resistance provided by the collagen sponges to replace the function of the lost tendon before its degradation and replacement with new extracellular matrix. This current study seeks to identify a biologically-derived scaffold with improved mechanical integrity that could be used in stem cell-based tissue engineered constructs for tendon repair.

Application of Tissue Engineering Techniques for Rotator Cuff Regeneration Using a Chitosan-Based Hyaluronan Hybrid Fiber Scaffold

2005 ◽  
Vol 33 (8) ◽  
pp. 1193-1201 ◽  
Author(s):  
Tadanao Funakoshi ◽  
Tokifumi Majima ◽  
Norimasa Iwasaki ◽  
Naoki Suenaga ◽  
Naohiro Sawaguchi ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Rotator Cuff ◽  
Mechanical Strength ◽  
Type I Collagen ◽  
Tangent Modulus ◽  
Rotator Cuff Tears ◽  
Type I ◽  
Scaffold Materials ◽  
Novel Scaffold

Background The current surgical procedures for irreparable rotator cuff tears have considerable limitations. Tissue engineering techniques using novel scaffold materials offer potential alternatives for managing these conditions. Hypothesis A chitosan-based hyaluronan hybrid scaffold could enhance type I collagen products with seeded fibroblasts and thereby increase the mechanical strength of regenerated tendon in vivo. Study Design Controlled laboratory study. Methods The scaffolds were created from chitosan-based hyaluronan hybrid polymer fibers. Forty-eight rabbit infraspinatus tendons and their humeral insertions were removed to create defects. Each defect was covered with a fibroblast-seeded scaffold (n = 16) or a non-fibroblast-seeded scaffold (n = 16). In the other 16 shoulders, the rotator cuff defect was left free as the control. At 4 and 12 weeks after surgery, the engineered tendons were assessed by histological, immunohistochemical (n = 2), and biomechanical (n = 6) analyses. Results Type I collagen was only seen in the fibroblast-seeded scaffold and increased in the regenerated tissue. The tensile strength and tangent modulus in the fibroblast-seeded scaffold were significantly improved from 4 to 12 weeks postoperatively. The fibroblast-seeded scaffold had a significantly greater tangent modulus than did the non-fibroblast-seeded scaffold and the control at 12 weeks. Conclusion This scaffold material enhanced the production of type I collagen and led to improved mechanical strength in the regenerated tissues of the rotator cuff in vivo. Clinical Relevance Rotator cuff regeneration is feasible using this tissue engineering technique.

Spontaneous Calcium Deposition in the Scaffolds from Human Dermal Solutions

2012 ◽  
Vol 506 ◽  
pp. 138-141
Author(s):  
K. Theerakittayakorn ◽  
T. Bunprasert
Keyword(s):  
Tissue Engineering ◽  
Type I Collagen ◽  
Collagen Matrix ◽  
Raw Material ◽  
Calcium Deposition ◽  
Type I ◽  
Human Dermis ◽  
Calcium Deposits ◽  
Scaffold Materials ◽  
Bovine Type

Human dermis was used as a new source of raw material for tissue engineering scaffold fabrication. Three human dermal solutions were prepared from different fractions after centrifugation and denoted as DS-1, DS-2 and DS-3. Approximately, the ratios of sulfated GAGs to collagen were 0.03, 0.02 and 0.04 for DS-1, DS-2 and DS-3, respectively. Scaffolds from the human dermal solutions and the commercial bovine type I collagen (Sigma®, St. Louis, MO, USA) were fabricated. The scaffolds were submerged in the normal culture medium and the calcium depositions were determined at day 1, 7 and 21. The highest calcium deposit was found in the scaffolds from type I collagen, the second were the scaffolds from DS-2, the third were the scaffolds from DS-1 and the lowest were the scaffolds from DS-3 for all time points. Histological sections stained with von Kossa stain explicitly exhibit the calcium depositions in the scaffolds. The calcium deposited in a manner according to the sulfated GAGs/collagen ratios of the scaffold materials. Calcium deposits are naturally incoperated into the collagen matrix of the human dermal solution-derived scaffolds. In bone tissue engineering, interpretation of experimental results should be careful of the spontaneous calcium deposition in scaffolds from collagen.

Chondrocytes behaviors within type I collagen microspheres and bulk hydrogels: an in vitro study

RSC Advances ◽  
2015 ◽  
Vol 5 (67) ◽  
pp. 54446-54453 ◽  
Author(s):  
Jun Liu ◽  
Hai Lin ◽  
Xiupeng Li ◽  
Yujiang Fan ◽  
Xingdong Zhang
Keyword(s):  
Tissue Engineering ◽  
Type I Collagen ◽  
In Vitro Study ◽  
Vitro Study ◽  
Type I ◽  
Design And Development ◽  
Proliferation And Differentiation ◽  
Cell Niche

Cell niche, which is considered to be critical to the proliferation and differentiation of cells, is one of the most important aspects for the design and development of ideal scaffolds in tissue engineering.

scholarly journals Efficacy of collagen and alginate hydrogels for the prevention of rat chondrocyte dedifferentiation

2018 ◽  
Vol 9 ◽  
pp. 204173141880243 ◽  
Author(s):  
Guang-Zhen Jin ◽  
Hae-Won Kim
Keyword(s):  
Tissue Engineering ◽  
Type I Collagen ◽  
Cartilage Tissue Engineering ◽  
Collagen Gel ◽  
Type Ii Collagen ◽  
Cartilage Tissue ◽  
Type I ◽  
Type Ii ◽  
Chondrogenic Phenotype

Dedifferentiation of chondrocytes remains a major problem in cartilage tissue engineering. The development of hydrogels that can preserve chondrogenic phenotype and prevent chondrocyte dedifferentiation is a meaningful strategy to solve dedifferentiation problem of chondrocytes. In the present study, three gels were prepared (alginate gel (Alg gel), type I collagen gel (Col gel), and their combination gel (Alg/Col gel)), and the in vitro efficacy of chondrocytes culture while preserving their phenotypes was investigated. While Col gel became substantially contracted with time, the cells encapsulated in Alg gel preserved the shape over the culture period of 14 days. The mechanical and cell-associated contraction behaviors of Alg/Col gel were similar to those of Alg. The cells in Alg and Alg/Col gels exhibited round morphology, whereas those in Col gel became elongated (i.e. fibroblast-like) during cultures. The cells proliferated with time in all gels with the highest proliferation being attained in Col gel. The expression of chondrogenic genes, including SOX9, type II collagen, and aggrecan, was significantly up-regulated in Alg/Col gel and Col gel, particularly in Col gel. However, the chondrocyte dedifferentiation markers, type I collagen and alkaline phosphatase ( ALP), were also expressed at significant levels in Col gel, which being contrasted with the events in Alg and Alg/Col gels. The current results suggest the cells cultured in hydrogels can express chondrocyte dedifferentiation markers as well as chondrocyte markers, which draws attention to choose proper hydrogels for chondrocyte-based cartilage tissue engineering.

scholarly journals Three-Dimensional Seeding of Chondrocytes Encapsulated in Collagen Gel into PLLA Scaffolds

2002 ◽  
Vol 11 (5) ◽  
pp. 489-494 ◽  
Author(s):  
Takashi Ushida ◽  
Katsuko Furukawa ◽  
Kenshi Toita ◽  
Tetsuya Tateishi
Keyword(s):  
Tissue Engineering ◽  
Type I Collagen ◽  
Collagen Gel ◽  
Type I ◽  
Cell Seeding ◽  
3D Scaffold ◽  
3D Scaffolds ◽  
Plla Scaffold ◽  
Collagen Solution

Tissue engineering approaches have been clinically tried to repair damaged articular cartilages. It is an essential step to uniformly seed chondrocytes into 3D scaffolds in order to reconstruct tissue-engineered cartilages in vitro, but the tissue engineering could not have been provided with efficient cell seeding methods. Type I collagen is clinically used and known as a cytocompatible material, having recognition sites for integrins. Collagen gel encapsulating chondrocytes has been tried for making regenerated cartilages, but it is found difficult to have the gel keep its original shape after long-term culture, because of shrinking. On the other hand, 3D scaffolds, either of a nonwoven structure or a sponge-like structure, involve difficulty in that chondrocytes could not be uniformly seeded, although they have adequate initial mechanical properties. In this study, by combining collagen gelation with a nonwoven PLLA scaffold, we achieved uniform cell seeding into the 3D scaffold. Bovine articular chondrocytes were mixed with type I collagen solution, and the solution was poured into the nonwoven PLLA scaffold (1.5 mm thick, f 15 mm). The collagen–chondrocyte mixture was made into gel at 37°C for 1 h. The 0.39% collagen mixture was viscous enough to prevent cells from precipitating during gelation. Almost all chondrocytes were able to be incorporated into the PLLA scaffolds by mixing with collagen solution and subsequently making into gel, while 30–40% of the chondrocytes seeded as a cell suspension were not trapped into the PLLA scaffolds. The method presented, where chondrocytes were mixed with collagen solution, and the mixture was incorporated into a 3D scaffold, then made into gel in the scaffold, could serve as an alternative for in vitro cartilage regeneration, also simultaneously having the advantages of both materials.

Collagen fibrillogenesis in vitro: interaction of types I and V collagen

1986 ◽  
Vol 44 ◽  
pp. 210-211
Author(s):  
Arthur J. Wasserman ◽  
Kathy C. Kloos ◽  
David E. Birk
Keyword(s):  
Type I Collagen ◽  
Corneal Stroma ◽  
Minor Component ◽  
Homogeneous Distribution ◽  
Type I ◽  
Type V Collagen ◽  
Fibril Morphology ◽  
Type V ◽  
In Vitro Interaction

Type I collagen is the predominant collagen in the cornea with type V collagen being a quantitatively minor component. However, the content of type V collagen (10-20%) in the cornea is high when compared to other tissues containing predominantly type I collagen. The corneal stroma has a homogeneous distribution of these two collagens, however, immunochemical localization of type V collagen requires the disruption of type I collagen structure. This indicates that these collagens may be arranged as heterpolymeric fibrils. This arrangement may be responsible for the control of fibril diameter necessary for corneal transparency. The purpose of this work is to study the in vitro assembly of collagen type V and to determine whether the interactions of these collagens influence fibril morphology.

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.

scholarly journals Restoration of Osteogenesis by CRISPR/Cas9 Genome Editing of the Mutated COL1A1 Gene in Osteogenesis Imperfecta

2021 ◽  
Vol 10 (14) ◽  
pp. 3141
Author(s):  
Hyerin Jung ◽  
Yeri Alice Rim ◽  
Narae Park ◽  
Yoojun Nam ◽  
Ji Hyeon Ju
Keyword(s):  
Osteogenesis Imperfecta ◽  
Type I Collagen ◽  
Disease Modeling ◽  
Bone Fragility ◽  
Osteogenic Potential ◽  
Type I ◽  
Gene Correction ◽  
Col1a1 Gene ◽  
Collagen Formation

Osteogenesis imperfecta (OI) is a genetic disease characterized by bone fragility and repeated fractures. The bone fragility associated with OI is caused by a defect in collagen formation due to mutation of COL1A1 or COL1A2. Current strategies for treating OI are not curative. In this study, we generated induced pluripotent stem cells (iPSCs) from OI patient-derived blood cells harboring a mutation in the COL1A1 gene. Osteoblast (OB) differentiated from OI-iPSCs showed abnormally decreased levels of type I collagen and osteogenic differentiation ability. Gene correction of the COL1A1 gene using CRISPR/Cas9 recovered the decreased type I collagen expression in OBs differentiated from OI-iPSCs. The osteogenic potential of OI-iPSCs was also recovered by the gene correction. This study suggests a new possibility of treatment and in vitro disease modeling using patient-derived iPSCs and gene editing with CRISPR/Cas9.

scholarly journals Effects of chitosan-collagen dressing on wound healing in vitro and in vivo assays

2021 ◽  
Vol 19 ◽  
pp. 228080002198969
Author(s):  
Min-Xia Zhang ◽  
Wan-Yi Zhao ◽  
Qing-Qing Fang ◽  
Xiao-Feng Wang ◽  
Chun-Ye Chen ◽  
...  
Keyword(s):  
Wound Healing ◽  
Wound Dressing ◽  
Type I Collagen ◽  
Inhibition Zone ◽  
Negative Control ◽  
Type I ◽  
Nih3t3 Cells ◽  
Post Operation

The present study was designed to fabricate a new chitosan-collagen sponge (CCS) for potential wound dressing applications. CCS was fabricated by a 3.0% chitosan mixture with a 1.0% type I collagen (7:3(w/w)) through freeze-drying. Then the dressing was prepared to evaluate its properties through a series of tests. The new-made dressing demonstrated its safety toward NIH3T3 cells. Furthermore, the CCS showed the significant surround inhibition zone than empty controls inoculated by E. coli and S. aureus. Moreover, the moisture rates of CCS were increased more rapidly than the collagen and blank sponge groups. The results revealed that the CCS had the characteristics of nontoxicity, biocompatibility, good antibacterial activity, and water retention. We used a full-thickness excisional wound healing model to evaluate the in vivo efficacy of the new dressing. The results showed remarkable healing at 14th day post-operation compared with injuries treated with collagen only as a negative control in addition to chitosan only. Our results suggest that the chitosan-collagen wound dressing were identified as a new promising candidate for further wound application.

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