In vitro comparison of human fibroblasts from intact and ruptured ACL for use in tissue engineering

Institut de Biologie et Chimie des Protéines, Centre National de la Recherche Scientifique - Université Claude Bernard Lyon 1, Lyon, France.
European cells & materials (Impact Factor: 4.89). 02/2007; 14:78-90; discussion 90-1.
Source: PubMed


The present study compares fibroblasts extracted from intact and ruptured human anterior cruciate ligaments (ACL) for creation of a tissue engineered ACL-construct, made of porcine small intestinal submucosal extracellular matrix (SIS-ECM) seeded with these ACL cells. The comparison is based on histological, immunohistochemical and RT-PCR analyses. Differences were observed between cells in a ruptured ACL (rACL) and cells in an intact ACL (iACL), particularly with regard to the expression of integrin subunits and smooth muscle actin (SMA). Despite these differences in the cell source, both cell populations behaved similarly when seeded on an SIS-ECM scaffold, with similar cell morphology, connective tissue organization and composition, SMA and integrin expression. This study shows the usefulness of naturally occurring scaffolds such as SIS-ECM for the study of cell behaviour in vitro, and illustrates the possibility to use autologous cells extracted from ruptured ACL biopsies as a source for tissue engineered ACL constructs.

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    • "ACLs contain a small subpopulation of fibroblasts positive for α-smooth muscle actin (α-SMA) [9], as well as cells that express certain markers of mesenchymal progenitor cells, including nucleostemin, SSEA-4, STRO-1, and Oct-4 [10-12]. "
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    ABSTRACT: Introduction Anterior cruciate ligament (ACL) degeneration is observed in most osteoarthritis (OA)-affected knee joints. However, the specific spatial and temporal relations of these changes and their association with extracellular matrix (ECM) degeneration are not well understood. The objective of this study was to characterize the patterns and relations of aging-related and OA-associated changes in ACL cells and the ECM. Methods Human knee joints from 80 donors (age 23 through 94) were obtained at autopsy. ACL degeneration was assessed histologically by using a quantitative scoring system. Tissue sections were analyzed for cell density, cell organization, ECM components, ECM-degrading enzymes and markers of differentiation, proliferation, and stem cells. Results Total cell number in normal ACL decreased with aging but increased in degenerated ACL, because of the formation of perivascular cell aggregates and islands of chondrocyte-like cells. Matrix metalloproteinase (MMP)-1, -3, and -13 expression was reduced in aging ACL but increased in degenerated ACL, mainly in the chondrocyte-like cells. Collagen I was expressed throughout normal and degenerated ACL. Collagen II and X were detected only in the areas with chondroid metaplasia, which also expressed collagen III. Sox9, Runt-related transcription factor 2 (Runx2), and scleraxis expression was increased in the chondrocyte-like cells in degenerated ACL. Alpha-smooth muscle actin (α-SMA), a marker of myofibroblasts and the progenitor cell marker STRO-1, decreased with aging in normal ACL. In degenerated ACL, the new cell aggregates were positive for α-SMA and STRO-1. Conclusions ACL aging is characterized by reduced cell density and activation. In contrast, ACL degeneration is associated with cell recruitment or proliferation, including progenitor cells or myofibroblasts. Abnormally differentiated chondrocyte-like cell aggregates in degenerated ACL produce abnormal ECM and may predispose to mechanical failure.
    Full-text · Article · Feb 2013 · Arthritis research & therapy
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    • "Another study compared the performance of fibroblasts extracted from both intact and ruptured human ACLs. They observed that cells extracted from the ruptured ACL were more useful in ligament tissue engineering [17]. Fibroblasts from other sources such as the skin are also being tested for their use in ligament tissue engineering. "
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    ABSTRACT: Tissue engineering is an emerging discipline that combines the principle of science and engineering. It offers an unlimited source of natural tissue substitutes and by using appropriate cells, biomimetic scaffolds, and advanced bioreactors, it is possible that tissue engineering could be implemented in the repair and regeneration of tissue such as bone, cartilage, tendon, and ligament. Whilst repair and regeneration of ligament tissue has been demonstrated in animal studies, further research is needed to improve the biomechanical properties of the engineered ligament if it is to play an important part in the future of human ligament reconstruction surgery. We evaluate the current literature on ligament tissue engineering and its role in anterior cruciate ligament reconstruction.
    Full-text · Article · Jan 2012
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    • "Brune et al. analyzed tendon cells from intact vs. torn ligament and observed differences in expression patterns but similar behaviour when seeded for tissue engineering (Brune et al., 2007). Other authors suggested fibroblast cultures from healing instead of intact tendon to be more appropriate for studying cellular tendon regeneration (Fu et al., 2008). "
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    ABSTRACT: Rotator cuff tears are common soft tissue injuries of the musculoskeletal system that heal by formation of repair tissue and may lead to high retear rates and joint dysfunction. In particular, tissue from chronic, large tendon tears is of such degenerative nature that it may be prone to retear after surgical repair. Besides several biomechanical approaches, biologically based strategies such as application of growth factors may be promising for increasing cell activity and production of extracellular tendon matrix at the tendon-to-bone unit. As a precondition for subsequent experimental growth factor application, the aim of the present study was to establish and characterize a human rotator cuff tendon cell culture. Long head biceps (LHB)- and supraspinatus muscle (SSP)- tendon samples from donor patients undergoing shoulder surgery were cultivated and examined at the RNA level for expression of collagen type-I, -II and -III, biglycan, decorin, tenascin-C, aggrecan, osteocalcin, tenomodulin and scleraxis (by Real-time PCR). Finally, results were compared to chondrocytes and osteoblasts as control cells. An expression pattern was found which may reflect a human rotator cuff tenocyte-like cell culture. Both SSP and LHB tenocyte-like cells differed from chondrocyte cell cultures in terms of reduced expression of collagen type-II (p<or=0.05) and decorin while higher levels of collagen type-I were seen (p<or=0.05). With respect to osteoblasts, tenocyte-like cells expressed lower levels of osteocalcin (p<or=0.05) as well as tenascin C, biglycan and collagen type-III. Expression of scleraxis, tenomodulin and aggrecan was similar between all cell types. This study represents a characterization of tenocyte-like cells from the human rotator cuff as close as possible. It helps analyzing their biological properties and allows further studies to improve production of tendon matrix and osteofibroblastic integration at the tendon-bone unit following tendon repair.
    Full-text · Article · Jul 2010 · European cells & materials
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