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.

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 Minispheroids as a Tool for Ligament Tissue Engineering: Do the Self-Assembly Techniques and Spheroid Dimensions Influence the Cruciate Ligamentocyte Phenotype?

2021 ◽  
Vol 22 (20) ◽  
pp. 11011
Author(s):  
Ingrid Zahn ◽  
Tobias Braun ◽  
Clemens Gögele ◽  
Gundula Schulze-Tanzil
Keyword(s):  
Gene Expression ◽  
Tissue Engineering ◽  
Self Assembly ◽  
Cruciate Ligament ◽  
Methyl Cellulose ◽  
Cell Viability Assay ◽  
Viability Assay ◽  
Extracellular Matrix Components ◽  
Collagen Type 1 ◽  
Ligament Tissue Engineering

Spheroid culture might stabilize the ligamentocyte phenotype. Therefore, the phenotype of lapine cruciate ligamentocyte (L-CLs) minispheroids prepared either by hanging drop (HD) method or by using a novel spheroid plate (SP) and the option of methyl cellulose (MC) for tuning spheroid formation was tested. A total of 250 and 1000 L-CLs per spheroid were seeded as HDs or on an SP before performing cell viability assay, morphometry, gene expression (qRT-PCR) and protein immunolocalization after 7 (HD/SP) and 14 (SP) days. Stable and viable spheroids of both sizes could be produced with both methods, but more rapidly with SP. MC accelerated the formation of round spheroids (HD). Their circular areas decreased significantly during culturing. After 7 days, the diameters of HD-derived spheroids were significantly larger compared to those harvested from the SP, with a tendency of lower circularity suggesting an ellipsoid shape. Gene expression of decorin increased significantly after 7 days (HD, similar trend in SP), tenascin C tended to increase after 7 (HD/SP) and 14 days (SP), whereas collagen type 1 decreased (HD/SP) compared to the monolayer control. The cruciate ligament extracellular matrix components could be localized in all mini-spheroids, confirming their conserved expression profile and their suitability for ligament tissue engineering.

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

Long-term hydrolytic degradation study on polymer-based embroidered scaffolds for ligament tissue engineering

2017 ◽  
Vol 47 (6) ◽  
pp. 1305-1320 ◽  
Author(s):  
Judith Hahn ◽  
Annette Breier ◽  
Harald Brünig ◽  
Gert Heinrich
Keyword(s):  
Tissue Engineering ◽  
Anterior Cruciate Ligament ◽  
Polylactic Acid ◽  
Cruciate Ligament ◽  
Hydrolytic Degradation ◽  
Ligament Injury ◽  
Degradation Behaviour ◽  
Native Anterior Cruciate Ligament ◽  
Anterior Cruciate ◽  
Ligament Tissue Engineering

Following anterior cruciate ligament injury, a mechanically stable tissue replacement is required for knee stability and to avoid subsequent damages. Tissue engineering of the anterior cruciate ligament demands a biocompatible scaffold with a controllable degradation profile to provide mechanical support for 3 to 6 months. It has been argued that embroidered textile scaffolds made of polylactic acid and poly(lactic-co-ɛ-caprolactone) fibres are a promising approach for the ligament tissue engineering with an adapted functionalization and cell seeding strategy. Therefore, the hydrolytic degradation behaviour of embroidered scaffolds made of polylactic acid and a combination of polylactic acid and poly(lactic-co-ɛ-caprolactone) fibres was investigated under physiological conditions for 168 days. The changes in the mechanical behaviour, the molecular weights as well as the surface structures were analysed. Sufficient mechanical properties comparable to native anterior cruciate ligament tissue could be demonstrated for scaffolds made of polylactic acid fibres after 6 months under hydrolysis. These results clarify the potential of using embroidered scaffolds for ligament tissue engineering.

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 Histochemical Characterisation and Gene Expression Analysis of Skeletal Muscles from Maremmana and Aubrac Steers Reared on Grazing and Feedlot Systems

Animals ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 656
Author(s):  
Giulia Foggi ◽  
Francesca Ciucci ◽  
Maria Conte ◽  
Laura Casarosa ◽  
Andrea Serra ◽  
...  
Keyword(s):  
Gene Expression ◽  
Physical Activity ◽  
Muscle Fibre ◽  
Skeletal Muscles ◽  
Gene Expression Analysis ◽  
Type I ◽  
Triceps Brachii ◽  
Gene Expressions ◽  
Muscle Properties ◽  
Voluntary Physical Activity

This study aimed to characterise the fibre composition of Triceps brachii (TB) and Semimembranosus (SM) muscles from 20 Maremmana (MA) and 20 Aubrac (AU) steers, and the effect of grazing activity in comparison with feedlot system. The histochemical method was performed with the m-ATPase method with an acid pre-incubation, thus allowing to distinguish type I, IIA, and IIB fibres. Additionally, on total RNA extracted from SM muscle, the expressions of atp1a1, mt-atp6, and capn1 genes were evaluated, in order to find potential associations with muscle fibre histochemical characteristics. In SM muscle, the MA steers had the greater frequency of oxidative fibres (type I and IIA) and the higher atp1a1 expression, in comparison to AU steers. Conversely, AU steers had a greater frequency of type IIB fibres, and the higher capn1 expression. A similar histochemical pattern was observed in TB muscle. The grazing activity was probably insufficient to determine differences both for fibre proportion and size, and gene expressions, except for mt-atp6 expression that was surprisingly highest in feedlot MA in comparison to other steers. These findings further the knowledge of muscle properties belonging to these breeds, and the effect of voluntary physical activity since few studies were available in this regard.

scholarly journals Arthroscopic Primary Repair of Proximal Anterior Cruciate Ligament Tears: With or Without Additional Suture Augmentation?

2018 ◽  
Vol 6 (7_suppl4) ◽  
pp. 2325967118S0006 ◽  
Author(s):  
Anne Jonkergouw ◽  
Jelle P. van der List ◽  
Gregory S. DiFelice
Keyword(s):  
Range Of Motion ◽  
Patient Reported Outcomes ◽  
Suture Anchor ◽  
Primary Repair ◽  
Cruciate Ligament ◽  
Type I ◽  
Acl Repair ◽  
Patient Reported ◽  
Anterior Cruciate

Objectives: Over the last years, arthroscopic primary repair of anterior cruciate ligament (ACL) tears has shown excellent results owing to appropriate patient selection (only repairing proximal ACL tears and good tissue quality), minimal invasive surgery (arthroscopy) and focus on early range of motion. Some surgeons have repaired proximal ACL tears without suture augmentation while others have used internal suture augmentation to reinforce and thus protect the repaired ligament during range of motion. No studies have yet compared the two surgical techniques. The objective of this study was to compare failure rates, reoperation rates and patient-reported outcomes of arthroscopic primary repair with versus without suture augmentation. Methods: A retrospective search for all patients treated with suture anchor arthroscopic primary ACL repair between April 2008 and June 2016 was performed. All patients with isolated proximal ACL tears (type I) were included. Since the development of internal suture augmentation, this reinforcement was added to the repaired ACLs. Minimum follow-up length was 1.0 years. Results: A total of 56 patients were included (mean age 33 years (range: 14 - 57), 59% male) of which 28 (50%) patients received additional suture augmentation. Mean follow-up was 2.3 years (range: 1.0-9.2). Six of all patients had reruptured their repaired ACL (10.7%), of which four underwent uncomplicated ACL reconstruction and two were treated conservatively. Four reruptures were initially treated with primary repair only (4/28, 14.3%) and two patients with additional suture augmentation (2/28, 7.1%; p = 0.431). During follow-up, three patients underwent reoperation (5.4%; two for medial meniscus tear (one in each group) and one for tibial suture anchor removal of the suture augmentation). Patient-reported outcomes have so far been collected in 20 patients without reruptures (currently collecting), with mean Lysholm score of 96, modified Cincinnati 94, SANE 93, pre-injury Tegner 6.7, postoperative Tegner 6.3 and subjective IKDC 91. Objective IKDC was A in 90%, B in 5%, C in 5%. Conclusion: In this study, the total failure rate of arthroscopic primary ACL repair was 10.7% and was lower with additional suture augmentation (7.1%) than primary repair alone (14.3%). Patients with failed ACL repair underwent uncomplicated primary ACL reconstruction. We recommend adding suture augmentation in high-risk patients (i.e. adolescents, ones with hyperlaxity, high contact sports), to protect the repaired ligament, especially during early range of motion. These data support treating type I proximal ACL tears with arthroscopic primary repair.

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