Establishment and comparison of fibroblast cell lines from the medial collateral and anterior cruciate ligaments of the rabbit

Ross, Stephen M.; Joshi, Rohit; Frank, Cyril B.

doi:10.1007/bf02624206

Cited by 52 publications

(42 citation statements)

References 10 publications

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“…ADMEM containing 5% FBS supported significant increases in collagen type I transcript expression for 6 of 8 groups tested versus their DMEM/10% FBS counterparts, attaining as much as a statistically significant 2-fold increase in (i) E > T and (ii) control test groups (p < 0.05, Table 2 fibroblasts in culture [21]; low collagen type 111 expression was therefore considered to be a favorable outcome in this study. The optimal expression by cells in culture with a mitogen followed by TGF-P placed [3] b > T in contention for further study.…”

Section: Discussionmentioning

confidence: 65%

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Growth factor induced fibroblast differentiation from human bone marrow stromal cells in vitro

Moreau

Chen

Bramono

et al. 2005

Journal Orthopaedic Research

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Utilizing a two-dimensional tissue culture plastic screening system and a fractional factorial design, specific media formulations and growth factor combinations were determined that support human bone marrow stromal cell (BMSC) differentiation toward fibroblast characteristics for utilization in tissue engineering, specifically cell morphology and alignment, metabolic activity, abundant expression of collagen types I and 111, and negligible expression of other tissue-specific markers. BMSCs were cultured for up to 14 days on tissue culture plastic, supplemented with Dulbecco's Minimal Essential Medium (DMEM)/IO% FBS or Advanced DMEM(ADMEM)/5% FBS. Each medium base was supplemented with one of nine possible growth factor combinations and ascorbate-2-phosphate (Asc-2-P) for the duration of culture. ADMEM supported comparable cell viability with half the serum content of the DMEM formulation. Asc-2-P was potent in promoting BMSC proliferation, in the absence of a mitogen, supporting significant increases in cell activity over 14 days of culture. DMEM promoted significant increases in cell viability for 7 of 9 growth factor groups when compared to their ADMEM counterparts. ADMEM, however, promoted increased cell transcript and protein expression, as 5 of 9 growth factor combinations induced a 200% increase in collagen type I versus equivalent DMEM cultures. Cell morphology and collagen type I immunostaining, when assessed in context of MTT and RNA results, identified 3 growth factor and medium combinations that supported fibroblast differentiation for future development of ligament tissue in vitro.

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Section: Discussionmentioning

confidence: 65%

“…The paper provides the first exhaustive screening of relevant biochemical factors alone, in combination and in sequential application to adult human BMSCs that optimally induce ligament fibroblast lineage. The results presented offer guidance to more complex future studies [10,12,21].…”

Section: J E Morivu Et Ui / Journul O F Orthopuerlicmentioning

confidence: 63%

Growth factor induced fibroblast differentiation from human bone marrow stromal cells in vitro

Moreau

Chen

Bramono

et al. 2005

Journal Orthopaedic Research

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“…Previous authors have reported minimal outgrowth ( G O cells in 6 days) from rabbit ACL explants [ 191 in serum-free media, and previous studies studying ACL explants and growth factors have been conducted in culture conditions of either 10% serum [33], or 0.5% serum [30]. It has been suggested that the addition of a small amount of serum to the explant culture supplies both the competence and the progression activity required to assay individual growth factors for ACL cells [28,30]. Although we attempted to demonstrate cell outgrowth from ACL explants in serum-free media in a pilot study for this experiment, we were unable to do so, and thus elected to supplement the media with 2% serum ~ a condition which may be more similar to that found inside the joint than a serum-free environment.…”

Section: Discussionmentioning

confidence: 99%

Cell outgrowth from the human ACL in vitro: regional variation and response to TGF‐β1

Murray

Bennett

Zhang

et al. 2002

Journal Orthopaedic Research

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One of the new methods being developed to stimulate healing of the human anterior cruciate ligament (ACL) after rupture is the implantation of a biodegradable scaffold which the host cells invade, populate and remodel. One of the cellular behaviors critical to the success of this method is cell outgrowth from the ligament remnants onto an adjacent scaffold. As morphological differences have been previously reported in the proximal and distal human ACL, the primary aim of this study was to determine if the cells from the proximal and distal ACL had different outgrowth behaviors as well. A second aim was to determine whether TGF-PI, reported to be a mitogen for fibroblasts, would affect the outgrowth behaviors from the proximal and distal ACL. To achieve these aims, explants from both the proximal and distal human ACL were placed into culture under various conditions and outgrowth measured for 42 days. In explants cultured with lo'%, fetal bovine serum (high serum group) the explants from the proximal ACL had an earlier start of outgrowth than the distal explants 0, < 0.015), and outgrowth rates were similar in the two groups. In explants cultured with 2% fetal bovine serum (low serum group), the explants from the proximal ACL had an earlier start to outgrowth 0, < 0.003) as well as a faster rate of outgrowth (p < 0.004) than the distal explants. The addition of TGF-PI to the low serum cultures significantly slowed the rate of outgrowth from both groups of ACL explants 0, < 0.025). These results suggest that outgrowth behaviors are different in the proximal and distal human ACL, and that TGB-PI has an inhibitory effect on cell outgrowth from ACL explants. However, this study is only a first step, and additional experiments are needed to further optimize tissue engineering parameters for enhancement of the repair of the ACL.

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“…12,16 Given similar shapes, larger ligaments with similar collagen composition and fibril size would be expected to be stronger than smaller ones. The ratio of T1C and T3C isoforms is one variable that contributes to differences in ligament scarring, healing, 9,11,13,17 and mechanical strength 15,17 among individuals and among different connective tissues (Figure 1).Collagen synthesis and remodeling follow the transcription of T1C and T3C messenger RNA (mRNA) and the conversion of procollagen into collagen fibers. 9,18-20 Extracellular matrix metalloproteinases and their inhibitors help to regulate collagen remodeling and play an important role in determining the composition and mechanical properties of the ACL.…”

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confidence: 99%

Sex, Collagen Expression, and Anterior Cruciate Ligament Strength in Rats

Romani

Langenberg

Belkoff

2010

Journal of Athletic Training

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Context: Sex-specific responses to steroid sex hormones have been suggested as a potential cause for the disparate anterior cruciate ligament (ACL) injury rates between male and female athletes. Type 1 collagen (T1C) and type 3 collagen (T3C) are crucial structural components that define the ligament's ability to withstand tensile loads. Messenger RNA (mRNA) is an important mediator of downstream collagen synthesis and remodeling, but the sex-specific mechanisms of collagen mRNA expression and ACL strength are unknown.Objective: To examine the influence of sex on T1C and T3C mRNA expression and mass-normalized stiffness and peak failure load in the ACLs of skeletally mature rats.Design: Observational study. Setting: Basic sciences and biomechanical testing laboratories.Patients or Other Participants: Nineteen 12-week-old male (n 5 9) and female (n 5 10) Sprague Dawley rats.Main Outcome Measure(s): We used real-time polymerase chain reaction to determine T1C and T3C mRNA expression and a hydraulic materials testing device to measure ACL stiffness and failure load. Nonparametric Wilcoxon rank sum tests were used to compare the groups.Results: Female rats had lower amounts of T3C mRNA expression and higher normalized ACL tangent stiffness and failure load than male rats.Conclusions: These findings suggest that sex-specific differences in T1C and T3C mRNA expression may play an important role in the downstream mechanical properties of the ACL.Key Words: knee injuries, women's health, real-time polymerase chain reaction Key Points N Compared with male rats, female rats demonstrated less type 3 collagen messenger RNA (mRNA) expression and greater normalized anterior cruciate ligament stiffness and load to failure.N Sex-specific differences in type 1 and type 3 collagen mRNA expression may influence the mechanical properties of the anterior cruciate ligament.T he anterior cruciate ligament (ACL) provides as much as 86% of the passive anterior restraint of the tibia on the femur. 1 As a result, ACL injuries often require surgical reconstruction and extensive rehabilitation to restore knee stability and ensure a full functional recovery. The long-term consequences of ACL injuries include accelerated knee joint degeneration; laxity; osteophyte formation; and type 2 collagen degradation, which may lead to early onset of osteoarthritis. [2][3][4][5] Most ACL injuries result from high tensile loads applied to the ligament during sudden changes of direction and jumping. 6 Whereas sex differences in landing and running biomechanics may place the ACL at risk, 7,8 the structure's ability to withstand those tensile loads likely depends upon several factors, including ligament size, rate of collagen remodeling, and collagen isoform. 9-11 Type 1 collagen (T1C) accounts for up to 87% 9,12 of the ligament's collagen content 9,12-14 and, along with type 3 collagen (T3C), acts to resist the loads applied to the ACL. 9,12 Type 3 collagen is also believed to play an important role in healing. [13][14][15] The extracellular matrix (ECM), ...

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Establishment and comparison of fibroblast cell lines from the medial collateral and anterior cruciate ligaments of the rabbit

Cited by 52 publications

References 10 publications

Growth factor induced fibroblast differentiation from human bone marrow stromal cells in vitro

Growth factor induced fibroblast differentiation from human bone marrow stromal cells in vitro

Cell outgrowth from the human ACL in vitro: regional variation and response to TGF‐β1

Sex, Collagen Expression, and Anterior Cruciate Ligament Strength in Rats

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