scholarly journals Maintenance of Ligament Homeostasis of Spheroid-Colonized Embroidered and Functionalized Scaffolds after 3D Stretch

2021 ◽  
Vol 22 (15) ◽  
pp. 8204
Author(s):  
Clemens Gögele ◽  
Jens Konrad ◽  
Judith Hahn ◽  
Annette Breier ◽  
Michaela Schröpfer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Suspension ◽  
Connexin 43 ◽  
Type I Collagen ◽  
Cruciate Ligament ◽  
Cyclic Stretch ◽  
Type I ◽  
Tenascin C ◽  
Gene Expression Control ◽  
Anterior Cruciate

Anterior cruciate ligament (ACL) ruptures are usually treated with autograft implantation to prevent knee instability. Tissue engineered ACL reconstruction is becoming promising to circumvent autograft limitations. The aim was to evaluate the influence of cyclic stretch on lapine (L) ACL fibroblasts on embroidered scaffolds with respect to adhesion, DNA and sulphated glycosaminoglycan (sGAG) contents, gene expression of ligament-associated extracellular matrix genes, such as type I collagen, decorin, tenascin C, tenomodulin, gap junctional connexin 43 and the transcription factor Mohawk. Control scaffolds and those functionalized by gas phase fluorination and cross-linked collagen foam were either pre-cultured with a suspension or with spheroids of LACL cells before being subjected to cyclic stretch (4%, 0.11 Hz, 3 days). Stretch increased significantly the scaffold area colonized with cells but impaired sGAGs and decorin gene expression (functionalized scaffolds seeded with cell suspension). Stretching increased tenascin C, connexin 43 and Mohawk but decreased decorin gene expression (control scaffolds seeded with cell suspension). Pre-cultivation of functionalized scaffolds with spheroids might be the more suitable method for maintaining ligamentogenesis in 3D scaffolds compared to using a cell suspension due to a significantly higher sGAG content in response to stretching and type I collagen gene expression in functionalized scaffolds.

scholarly journals Cruciate Ligament Cell Sheets Can Be Rapidly Produced on Thermoresponsive poly(glycidyl ether) Coating and Successfully Used for Colonization of Embroidered Scaffolds

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 877
Author(s):  
Ingrid Zahn ◽  
Daniel David Stöbener ◽  
Marie Weinhart ◽  
Clemens Gögele ◽  
Annette Breier ◽  
...  
Keyword(s):  
Gene Expression ◽  
Type I Collagen ◽  
Cruciate Ligament ◽  
Glycidyl Ether ◽  
Stable Gene ◽  
Type I ◽  
Cell Sheets ◽  
Thermoresponsive Polymers ◽  
Related Phenotype ◽  
Anterior Cruciate

Anterior cruciate ligament (ACL) cell sheets combined with biomechanically competent scaffolds might facilitate ACL tissue engineering. Since thermoresponsive polymers allow a rapid enzyme-free detachment of cell sheets, we evaluated the applicability of a thermoresponsive poly(glycidyl ether) (PGE) coating for cruciate ligamentocyte sheet formation and its influence on ligamentocyte phenotype during sheet-mediated colonization of embroidered scaffolds. Ligamentocytes were seeded on surfaces either coated with PGE or without coating. Detached ligamentocyte sheets were cultured separately or wrapped around an embroidered scaffold made of polylactide acid (PLA) and poly(lactic-co-ε-caprolactone) (P(LA-CL)) threads functionalized by gas-phase fluorination and with collagen foam. Ligamentocyte viability, protein and gene expression were determined in sheets detached from surfaces with or without PGE coating, scaffolds seeded with sheets from PGE-coated plates and the respective monolayers. Stable and vital ligamentocyte sheets could be produced within 24 h with both surfaces, but more rapidly with PGE coating. PGE did not affect ligamentocyte phenotype. Scaffolds could be colonized with sheets associated with high cell survival, stable gene expression of ligament-related type I collagen, decorin, tenascin C and Mohawk after 14 d and extracellular matrix (ECM) deposition. PGE coating facilitates ligamentocyte sheet formation, and sheets colonizing the scaffolds displayed a ligament-related phenotype.

scholarly journals Gene Expression of Type I and Type III Collagen by Mechanical Stretch in Anterior Cruciate Ligament Cells

2002 ◽  
Vol 27 (3) ◽  
pp. 139-144 ◽  
Author(s):  
Sung-Gon Kim ◽  
Toshihiro Akaike ◽  
Tadashi Sasagawa ◽  
Yoriko Atomi ◽  
Hisashi Kurosawa
Keyword(s):  
Gene Expression ◽  
Anterior Cruciate Ligament ◽  
Cruciate Ligament ◽  
Mechanical Stretch ◽  
Type Iii Collagen ◽  
Type I ◽  
Type Iii ◽  
Anterior Cruciate

scholarly journals Torn ACL: A New Bioengineered Substitute Brought from the Laboratory to the Knee Joint

2004 ◽  
Vol 1 (2) ◽  
pp. 115-121 ◽  
Author(s):  
Francine Goulet ◽  
Denis Rancourt ◽  
Réjean Cloutier ◽  
Pierrot Tremblay ◽  
Anne-Marie Belzil ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Cruciate Ligament ◽  
New Technology ◽  
Collagen Matrix ◽  
Type I ◽  
Acl Injuries ◽  
Fully Integrated ◽  
Post Surgery ◽  
The Usa ◽  
Anterior Cruciate

Anterior cruciate ligament (ACL) injuries occur at an annual rate of 120 000 in the USA, and many need reconstructive surgery. We report successful results at 1–13 months following implantation of bioengineered ACL (bACL) in goats. A bACL has been developed using autologous ACL cells, a collagen matrix and bone plugs. The extremities of the bACL were fully integrated into the femur and tibia of the host. Vascularisation of the grafts was extensive 1 month post-surgery and improved with time. At 6 months post-grafting, histological and ultrastructural observations demonstrated a highly organised ligamentous structure, rich in type I collagen fibres and fibroblasts. At the implants' insertion sites, characteristic fibrocartilage was observed having well aligned chondrocytes and collagen fibrils. After a year, mechanical rupture of the grafts demonstrated a major gain in strength. Eventual applications of this new technology in humans include multiple uses in orthopaedic, dental and reconstructive surgeries.

Induction of vascular smooth muscle cell tenascin-C gene expression by denatured type I collagen is dependent upon a beta3 integrin-mediated mitogen-activated protein kinase pathway and a 122-base pair promoter element

1999 ◽  
Vol 112 (4) ◽  
pp. 435-445 ◽  
Author(s):  
P.L. Jones ◽  
F.S. Jones ◽  
B. Zhou ◽  
M. Rabinovitch
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Smooth Muscle Cell ◽  
Muscle Cell ◽  
Type I Collagen ◽  
Type I ◽  
C Protein ◽  
Tenascin C ◽  
Beta3 Integrin

Tenascin-C is an extracellular matrix glycoprotein, the expression of which is upregulated in remodeling arteries. In previous studies we showed that the presence of tenascin-C alters vascular smooth muscle cell shape and amplifies their proliferative response by promoting growth factor receptor clustering and phosphorylation. Moreover, we demonstrated that denatured type I collagen induces smooth muscle cell tenascin-C protein production via beta3 integrins. In the present study, we examine the pathway by which beta3 integrins stimulate expression of tenascin-C, and define a promoter sequence that is critical for its induction. On native collagen, A10 smooth muscle cells adopt a stellate morphology and produce low levels of tenascin-C mRNA and protein, whereas on denatured collagen they spread extensively and produce high levels of tenascin-C mRNA and protein, which is incorporated into an elaborate extracellular matrix. Increased tenascin-C synthesis on denatured collagen is associated with elevated protein tyrosine phosphorylation, including activation of extracellular signal-regulated kinases 1 and 2 (ERK1 and ERK2). beta3 integrin function-blocking antibodies attenuate ERK1/2 activation and tenascin-C protein synthesis. Consistent with these findings, treatment with the specific MEK inhibitor, PD 98059, results in suppression of tenascin-C protein synthesis. To investigate whether beta3 integrin-dependent activation of ERK1/2 regulates the tenascin-C promoter, we transfected A10 cells with a full-length (approx. 4 kb) mouse tenascin-C gene promoter-chloramphenicol acetyltransferse reporter construct and showed that, relative to native collagen, its activity is increased on denatured collagen. Next, to identify regions of the promoter involved, we examined a series of tenascin-C promoter constructs with 5′ deletions and showed that denatured collagen-dependent promoter activity was retained by a 122-base pair element, located -43 to -165 bp upstream of the RNA start site. Activation of this element was suppressed either by blocking beta3 integrins, or by preventing ERK1/2 activation. These observations demonstrate that smooth muscle cell binding to beta3 integrins activates the mitogen activated protein kinase pathway, which is required for the induction of tenascin-C gene expression via a potential extracellular matrix response element in the tenascin-C gene promoter. Our data suggest a mechanism by which remodeling of type I collagen modulates tenascin-C gene expression via a beta3 integrin-mediated signaling pathway, and as such represents a paradigm for vascular development and disease whereby smooth muscle cells respond to perturbations in extracellular matrix composition by altering their phenotype and patterns of gene expression.

Gap junctions in IL-1β-mediated cell survival response to strain

2011 ◽  
Vol 110 (5) ◽  
pp. 1425-1431 ◽  
Author(s):  
Jie Qi ◽  
Liqun Chi ◽  
Donald Bynum ◽  
Albert J. Banes
Keyword(s):  
Gene Expression ◽  
Cell Death ◽  
Cell Survival ◽  
Connexin 43 ◽  
Type I Collagen ◽  
Cyclic Strain ◽  
Marker Genes ◽  
Type I ◽  
Mechanical Stimuli

Mechanical stimuli play important roles in proliferation and differentiation of connective tissue cells, and development and homeostatic maintenance of tissues. However, excessive mechanical loading to a tissue can injure cells and disrupt the matrix, as occurs in tendinopathy. Tendinopathy is a common clinical problem in athletes and in many occupational settings due to overuse of the tendon. Moreover, interleukin (IL)-1β is generally considered to be a “bad” cytokine, activating NF-κb and cell death and inducing matrix metalloproteinase (MMPs 1, 2, 3) expression and matrix destruction. However, activated NF-κB can also drive a cell survival pathway. We have reported that cyclic strain induced tenocyte death in three-dimensional (3D) cultures, and IL-1β could promote cell survival under strain. Therefore, it was hypothesized that 1) cyclic strain could induce cell death in tenocytes as observed in pathologic tendons in vivo; 2) a gene expression profile indicative of tendinopathy could be identified; and 3) low-dose IL-1β could protect cells from strain-induced, tendinopathy-like changes. Human tenocytes were cultured in 3D type I collagen hydrogels and subjected to 3.5% elongation at 1 Hz for 1 h/day for up to 5 days with or without IL-1β. Real-time RT-PCR data showed that cyclic strain regulated the expression of tendinopathy marker genes in a manner similar to that found in pathological tendons from patients and that addition of IL-1β reversed the gene expression changes to control levels. Results of further studies showed that IL-1β may modulate cell survival through upregulating the expression of connexin 43, which is involved in the modulation of cell death/survival in a variety of cells and tissues. The elucidation of the mechanisms underlying strain-induced cell death and recovery from strain injury will facilitate our understanding of the pathogenesis of tendinopathy and may lead to the discovery of new molecular targets for early diagnosis and treatment of tendinopathy.

Effects of Wavy Microgroove Structure on Ligament Fibroblast Cell and Nuclear Morphology

2013 ◽  
Author(s):  
Chen-Hsiu Sung ◽  
Pen-hsiu Grace Chao
Keyword(s):  
Type I Collagen ◽  
Cruciate Ligament ◽  
Fibroblast Cell ◽  
The Body ◽  
Type I ◽  
Fibrous Materials ◽  
Cell Behaviors ◽  
Substrate Structure ◽  
Wavy Structure ◽  
Anterior Cruciate

Type I collagen, the most abundant extracellular matrix component in the body, exists as fibers that are organized in wavy parallel fibers in many tissues [1, 2]. Cells embedded within this wavy pattern exhibit a 3D morphology that undulates with the fibers. The phenomenon that cell morphology follows substrate structure, such as cell elongation when attached to fibrous materials, is known as contact guidance [3]. As the cytoskeleton supports cellular structures and mediate numerous intracellular processes, changes to the cytoskeleton structures lead to modified cell behaviors and even fate [4, 5]. Studies have shown that fibroblasts express more ligament phenotypic markers when cultured on straight micropatterns or aligned fibers [6, 7]. However, few studies have investigated the influences of the wavy structures. To characterize the effects of the wavy structure, anterior cruciate ligament (ACL) fibroblasts were seeded in wavy microgrooves and their morphological responses were analyzed.

scholarly journals A Novel Approach to Augmenting Allograft Hamstring Anterior Cruciate Ligament Reconstructions Utilizing a Resorbable Type I Collagen Matrix with Platelet Rich Plasma

2021 ◽  
Vol 2021 ◽  
pp. 1-6
Author(s):  
Sean Mc Millan ◽  
Danielle Thorn ◽  
Elizabeth Ford
Keyword(s):  
Anterior Cruciate Ligament ◽  
Type I Collagen ◽  
Platelet Rich Plasma ◽  
Cruciate Ligament ◽  
Collagen Matrix ◽  
Type I ◽  
Biologic Graft ◽  
Graft Incorporation ◽  
Novel Approach ◽  
Anterior Cruciate

Background. Anterior cruciate ligament reconstruction (ACLR) is one of the most common lower extremity orthopedic surgeries performed in the United States. Annually, between 100,000 and 200,000 ACL tears affect 1 in 3,000 people. The selection of autograft versus allograft for ACLR has been widely discussed in terms of risk of graft failure. Allograft reconstructions have been shown to have higher rerupture rates. One factor contributing to this risk is delayed biologic graft incorporation. Methods. A retrospective review was performed examining 14 patients who underwent an augmented quadruple-stranded hamstring allograft ACLR with a type I resorbable collagen matrix impregnated with platelet-rich plasma (PRP). Results. Within our clinical practice, the augmentation of quadruple-stranded hamstring allograft ACLR with a type I resorbable matrix impregnated with PRP has yielded good early clinical success at 2-year outcomes ( N = 14 ). Zero ACLR failures have been reported to date in this series. Conclusion. This case series offers a novel approach for soft tissue allograft ACLR augmented with a type I collagen matrix impregnated with PRP. The authors theorize that this augmentation may improve biologic graft incorporation into the host bone tunnels.

scholarly journals Enhanced Growth of Lapine Anterior Cruciate Ligament-Derived Fibroblasts on Scaffolds Embroidered from Poly(l-lactide-co-ε-caprolactone) and Polylactic Acid Threads Functionalized by Fluorination and Hexamethylene Diisocyanate Cross-Linked Collagen Foams

2020 ◽  
Vol 21 (3) ◽  
pp. 1132 ◽  
Author(s):  
Clemens Gögele ◽  
Judith Hahn ◽  
Cindy Elschner ◽  
Annette Breier ◽  
Michaela Schröpfer ◽  
...  
Keyword(s):  
Gene Expression ◽  
Tissue Engineering ◽  
Polylactic Acid ◽  
Cruciate Ligament ◽  
Donor Site ◽  
Hexamethylene Diisocyanate ◽  
Type I ◽  
Gene Expressions ◽  
Extracellular Matrix Components ◽  
Anterior Cruciate

Reconstruction of ruptured anterior cruciate ligaments (ACLs) is limited by the availability and donor site morbidity of autografts. Hence, a tissue engineered graft could present an alternative in the future. This study was undertaken to determine the performance of lapine (L) ACL-derived fibroblasts on embroidered poly(l-lactide-co-ε-caprolactone) (P(LA-CL)) and polylactic acid (PLA) scaffolds in regard to a tissue engineering approach for ACL reconstruction. Surface modifications of P(LA-CL)/PLA by gas-phase fluorination and cross-linking of a collagen foam using either ethylcarbodiimide (EDC) or hexamethylene diisocyanate (HMDI) were tested regarding their influence on cell adhesion, growth and gene expression. The experiments were performed using embroidered P(LA-CL)/PLA scaffolds that were seeded dynamically or statically with LACL-derived fibroblasts. Scaffold cytocompatibility, cell survival, numbers, metabolic activity, ultrastructure and sulfated glycosaminoglycan (sGAG) synthesis were evaluated. Quantitative real-time polymerase chain reaction (QPCR) revealed gene expression of collagen type I (COL1A1), decorin (DCN), tenascin C (TNC), Mohawk (MKX) and tenomodulin (TNMD). All tested scaffolds were highly cytocompatible. A significantly higher cellularity and larger scaffold surface areas colonized by cells were detected in HMDI cross-linked and fluorinated scaffolds compared to those cross-linked with EDC or without any functionalization. By contrast, sGAG synthesis was higher in controls. Despite the fact that the significance level was not reached, gene expressions of ligament extracellular matrix components and differentiation markers were generally higher in fluorinated scaffolds with cross-linked collagen foams. LACL-derived fibroblasts maintained their differentiated phenotype on fluorinated scaffolds supplemented with a HMDI cross-linked collagen foam, making them a promising tool for ACL tissue engineering.

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