Evaluation of Cartilage Specific Matrix Synthesis of Human Articular Chondrocytes after Extended Propagation on Microcarriers by Image Analysis

Goepfert, Christine; Lutz, Vivien; Lünse, Svenja; Kittel, Sabrina; Wiegandt, Katharina; Kammal, Michael; Püschel, Klaus; Pörtner, Ralf

doi:10.1177/039139881003300405

Cited by 16 publications

(9 citation statements)

References 49 publications

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“…5). Some growth factors included in PRP, such as TGF-b1, IGF-I, BMP-2 and -7, have already been reported to have an effect of promoting collagen synthesis 38,39 , which supports our findings that PRP enhances collagen synthesis. This increase of collagen synthesis could represent another mechanism by which PRP enhances AC healing.…”

Section: Discussionsupporting

confidence: 91%

The effect of platelet-rich plasma on the regenerative therapy of muscle derived stem cells for articular cartilage repair

Mifune

Matsumoto

Takayama

et al. 2013

Osteoarthritis and Cartilage

141

112

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Section: Discussionsupporting

confidence: 91%

The effect of platelet-rich plasma on the regenerative therapy of muscle derived stem cells for articular cartilage repair

Mifune

Matsumoto

Takayama

et al. 2013

Osteoarthritis and Cartilage

141

112

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“…These activated chondrocytes secrete not only the matrix molecules typical for chondrocytes but also start secreting the molecules similar to the osteogenic cells of bone. Goepfert et al (2010) also stated that proliferating chondrocytes are known to undergo dedifferentiation characterized by down-regulation of collagen type II and proteoglycan production, and by upregulation of collagen type I synthesis. Re-expression of cartilage specific matrix components by proliferating chondrocytes is therefore critical for successful cartilage repair.…”

Section: Discussionmentioning

confidence: 99%

Immunohistochemical analysis of ageing and osteoarthritic articular cartilage

et al. 2010

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Articular cartilage degeneration seen in osteoarthritis is primarily the consequence of events within the articular cartilage that leads to the production of proteases by chondrocytes. 22 osteoarthritic cartilage specimens were obtained from patients with primary osteoarthritis (46-81 years) undergoing total knee replacement. 12 age-matched (41-86 years) and 16 young (16-40 years) non-osteoarthritic control cartilage specimens were obtained from the cadavers in the department of Anatomy and from patients undergoing lower limb amputation in Trauma center of PGIMER, Chandigarh. 5 μ thick paraffin sections were stained for osteocalcin, osteopontin, osteonectin and alkaline phosphatase to analyze their expression in hypertrophied chondrocytes and osteoarthritic cartilage matrix and to compare the staining intensity with that of normal ageing articular cartilage. Immunohistochemical staining of tissue sections revealed moderate to strong cytoplasmic staining for all four stains in all the specimens of the osteoarthritic group compared to age-matched control. The immunohistochemical scores were significantly higher in the osteoarthritic group for all four stains. The features of the osteoarthritic articular cartilage were markedly different from the non-osteoarthritic age-matched articular cartilage suggesting that osteoarthritis is not an inevitable feature of aging.

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“…Based on all mentioned it is clear that chondrocyte isolation from an abundant source with a high yield, together with an effective and cheap preliminary phenotype confirmation method, would be greatly beneficial to boost the development of cartilage tissue engineering (Cetinkaya et al, 2011; Goepfert et al, 2010; Schrobback et al, 2011). This study was therefore designed to provide a relatively simple, yet effective procedure for isolation and culturing of human tissue derived primary chondrocytes up to the second passage.…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

Naranda

Gradišnik

Gorenjak

et al. 2017

PeerJ

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BackgroundCartilage tissue engineering is a fast-evolving field of biomedical engineering, in which the chondrocytes represent the most commonly used cell type. Since research in tissue engineering always consumes a lot of cells, simple and cheap isolation methods could form a powerful basis to boost such studies and enable their faster progress to the clinics. Isolated chondrocytes can be used for autologous chondrocyte implantation in cartilage repair, and are the base for valuable models to investigate cartilage phenotype preservation, as well as enable studies of molecular features, nature and scales of cellular responses to alterations in the cartilage tissue.MethodsIsolation and consequent cultivation of primary human adult articular chondrocytes from the surgical waste obtained during total knee arthroplasty (TKA) was performed. To evaluate the chondrogenic potential of the isolated cells, gene expression of collagen type 2 (COL2), collagen 1 (COL1) and aggrecan (ACAN) was evaluated. Immunocytochemical staining of all mentioned proteins was performed to evaluate chondrocyte specific production.ResultsCartilage specific gene expression of COL2 and ACAN has been shown that the proposed protocol leads to isolation of cells with a high chondrogenic potential, possibly even specific phenotype preservation up to the second passage. COL1 expression has confirmed the tendency of the isolated cells dedifferentiation into a fibroblast-like phenotype already in the second passage, which confirms previous findings that higher passages should be used with care in cartilage tissue engineering. To evaluate the effectiveness of our approach, immunocytochemical staining of the evaluated chondrocyte specific products was performed as well.DiscussionIn this study, we developed a protocol for isolation and consequent cultivation of primary human adult articular chondrocytes with the desired phenotype from the surgical waste obtained during TKA. TKA is a common and very frequently performed orthopaedic surgery during which both femoral condyles are removed. The latter present the ideal source for a simple and relatively cheap isolation of chondrocytes as was confirmed in our study.

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Evaluation of Cartilage Specific Matrix Synthesis of Human Articular Chondrocytes after Extended Propagation on Microcarriers by Image Analysis

Abstract: Expanding human articular chondrocytes on microcarriers omitting subcultivation steps leads to superior ratios of collagen type II to type I forming cells compared to the expansion in conventional monolayer culture.

Cited by 16 publications

References 49 publications

The effect of platelet-rich plasma on the regenerative therapy of muscle derived stem cells for articular cartilage repair

The effect of platelet-rich plasma on the regenerative therapy of muscle derived stem cells for articular cartilage repair

Immunohistochemical analysis of ageing and osteoarthritic articular cartilage

Isolation and characterization of human articular chondrocytes from surgical waste after total knee arthroplasty (TKA)

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