The synovial microenvironment of osteoarthritic joints alters RNA-seq expression profiles of human primary articular chondrocytes

Lewallen, Eric A.; Bonin, Carolina A.; Li, Xin; Smith, Jay; Karperien, Marcel; Larson, A. Noelle; Lewallen, David G.; Cool, Simon M.; Krych, Aaron J.; Leontovich, Alexey A.; Im, Hee Jeong; Wijnen, André J. van

doi:10.1016/j.gene.2016.06.063

Cited by 17 publications

(19 citation statements)

References 51 publications

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“…However, RNA-seq has the unique ability to determine the relative expression levels of all genes within a cell or tissue. This technique has not yet been widely used on articular cartilage, [8][9][10][11][12][13][14] and to the best of our knowledge, the transcriptome-wide profile of articular cartilage obtained from healthy knee joints has not been reported.…”

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confidence: 99%

Transcriptional profiling of articular cartilage in a porcine model of early post‐traumatic osteoarthritis

Sieker

Proffen

Waller

et al. 2017

Journal Orthopaedic Research

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To identify the molecular pathophysiology present in early post-traumatic osteoarthritis (PTOA), the transcriptional profile of articular cartilage and its response to surgical PTOA induction were determined. Thirty six Yucatan minipigs underwent anterior cruciate ligament (ACL) transection and were randomly assigned in equal numbers to no further treatment, reconstruction or ligament repair. Cartilage was harvested at 1 and 4 weeks post-operatively and histology and RNA-sequencing were performed and compared to controls. Microscopic cartilage scores significantly worsened at 1 (p = 0.028) and 4 weeks (p = 0.001) post-surgery relative to controls, but did not differ between untreated, reconstruction or repair groups. Gene expression after ACL reconstruction and ACL transection were similar, with only 0.03% (including SERPINB7 and CR2) and 0.2% of transcripts (including INHBA) differentially expressed at 1 and 4 weeks respectively. COL2A1, COMP, SPARC, CHAD, and EF1ALPHA were the most highly expressed non ribosomal, non mitochondrial genes in the controls and remained abundant after surgery. A total of 1,275 genes were differentially expressed between 1 and 4 weeks post-surgery. With the treatment groups pooled, 682 genes were differentially expressed at both time-points, with the most significant changes observed in MMP1, COCH, POSTN, CYTL1, and PTGFR. This study confirmed the development of a microscopic PTOA stage after ACL surgery in the porcine model. Upregulation of multiple proteases (including MMP1 and ADAMTS4) were found; however, the level of expression remained orders of magnitude below that of extracellular matrix protein-coding genes (including COL2A1 and ACAN). In summary, genes with established roles in PTOA as well as novel targets for specific intervention were identified. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:318-329, 2018.

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confidence: 99%

Transcriptional profiling of articular cartilage in a porcine model of early post‐traumatic osteoarthritis

Sieker

Proffen

Waller

et al. 2017

Journal Orthopaedic Research

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“…Furthermore, considering alginate beads model, we demonstrated that GAGs production was significantly higher in NC cultures compared to ACs and we conducted the cell culture until obtaining a quantifiable cartilage matrix (seven weeks). Different authors confirmed that NCs show superior and more reproducible in vitro chondrogenic potential [7,18,19,20]; this is possibly due to the pathological condition of articular tissues that are commonly harvested in patients with osteoarthritis or after sport trauma (during joint replacement surgery) [21,22]. These patients were typically treated with cortisone and other anti-inflammatory drugs, which are able to partially suppress the inflammatory cascade but, at the same time, could have an inhibition effect on articular chondrocyte proliferation.…”

Section: Discussionmentioning

confidence: 99%

Human Engineered Cartilage and Decellularized Matrix as an Alternative to Animal Osteoarthritis Model

et al. 2018

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(1) Objective: to obtain a reproducible, robust, well-defined, and cost-affordable in vitro model of human cartilage degeneration, suitable for drug screening; (2) Methods: we proposed 3D models of engineered cartilage, considering two human chondrocyte sources (articular/nasal) and five culture methods (pellet, alginate beads, silk/alginate microcarriers, and decellularized cartilage). Engineered cartilages were treated with pro-inflammatory cytokine IL-1β to promote cartilage degradation; (3) Results: articular chondrocytes have been rejected since they exhibit low cellular doubling with respect to nasal cells, with longer culture time for cell expansion; furthermore, pellet and alginate bead cultures lead to insufficient cartilage matrix production. Decellularized cartilage resulted as good support for degeneration model, but long culture time and high cell amount are required to obtain the adequate scaffold colonization. Here, we proposed, for the first time, the combined use of decellularized cartilage, as aggrecanase substrate, with pellet, alginate beads, or silk/alginate microcarriers, as polymeric scaffolds for chondrocyte cultures. This approach enables the development of suitable models of cartilaginous pathology. The results obtained after cryopreservation also demonstrated that beads and microcarriers are able to preserve chondrocyte functionality and metabolic activity; (4) Conclusions: alginate and silk/alginate-based scaffolds can be easily produced and cryopreserved to obtain a cost-affordable and ready-to-use polymer-based product for the subsequent screening of anti-inflammatory drugs for cartilage diseases.

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“…Because human adipose tissue is a readily accessible source of such multipotent adult stem cells, human adipose‐deriveds MSCs have been well characterized and are being considered for stem cell treatments of musculoskeletal tissues disorders . Our research group has extensively studied a number of joint and cartilage disorders that could benefit from MSC therapy as well as examined environmental factors that may impact the effective therapeutic dose of stem cells …”

Section: Introductionmentioning

confidence: 99%

“…18,19 Because human adipose tissue is a readily accessible source of such multipotent adult stem cells, human adipose-deriveds MSCs have been well characterized and are being considered for stem cell treatments of musculoskeletal tissues disorders. 20,21 Our research group has extensively studied a number of joint and cartilage disorders that could benefit from MSC therapy [22][23][24][25][26][27] as well as examined environmental factors that may impact the effective therapeutic dose of stem cells. [28][29][30][31][32][33][34] Injected local anesthetics are commonly applied for both diagnostic and therapeutic purposes in perioperative and ambulatory settings in orthopedic practices 35 and for the treatment of musculoskeletal pain.…”

Section: Introductionmentioning

confidence: 99%

Effect of Lidocaine on Viability and Gene Expression of Human Adipose–derived Mesenchymal Stem Cells: An in vitro Study

Nie

Kubrova

et al. 2019

PM&R

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Objective To assess the biologic effects of lidocaine on the viability, proliferation, and function of human adipose tissue–derived mesenchymal stromal/stem cells (MSCs) in vitro. Methods Adipose‐derived MSCs from three donors were exposed to lidocaine at various dilutions (2 mg/mL to 8 mg/mL) and exposure times (0.5 to 4 hours). Cell number and viability, mitochondrial activity, and real‐time reverse‐transcriptase quantitative polymerase chain reaction (RT‐qPCR) were analyzed at 0 (immediate effects) or 24 and 48 hours (recovery effects) after treatment with lidocaine. Results Trypan blue staining showed that increasing concentrations of lidocaine decreased the number of observable viable cells. 3‐[4,5,dimethylthiazol‐2‐yl]‐5‐[3‐carboxymethoxy‐phenyl]‐2‐[4‐sulfophenyl]‐2H‐tetrazolium (MTS) assays revealed a concentration‐ and time‐ dependent decline of mitochondrial activity and proliferative ability. Gene expression analysis by RT‐qPCR revealed that adipose‐derived MSCs exposed to lidocaine express robust levels of stress response/cytoprotective genes. However, higher concentrations of lidocaine caused a significant downregulation of these genes. No significant differences were observed in expression of extracellular matrix (ECM) markers COL1A1 and DCN except for COL3A1 (P < .05). Levels of messenger RNA (mRNA) for proliferation markers (CCNB2, HIST2H4A, P < .001) and MKI67 (P < .001) increased at 24 and 48 hours. Expression levels of several transcription factors— including SP1, PRRX1, and ATF1—were modulated in the same manner. MSC surface markers CD44 and CD105 demonstrated decreased expression immediately after treatment, but at 24 and 48 hours postexposure, the MSC markers showed no significant difference among groups. Conclusion Lidocaine is toxic to MSCs in a dose‐ and time‐ dependent manner. MSC exposure to high (4‐8 mg/mL) concentrations of lidocaine for prolonged periods can affect their biologic functions. Although the exposure time in vivo is short, it is essential to choose safe concentrations when applying lidocaine along with MSCs to avoid compromising the viability and potency of the stem cell therapy.

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The synovial microenvironment of osteoarthritic joints alters RNA-seq expression profiles of human primary articular chondrocytes

Cited by 17 publications

References 51 publications

Transcriptional profiling of articular cartilage in a porcine model of early post‐traumatic osteoarthritis

Transcriptional profiling of articular cartilage in a porcine model of early post‐traumatic osteoarthritis

Human Engineered Cartilage and Decellularized Matrix as an Alternative to Animal Osteoarthritis Model

Effect of Lidocaine on Viability and Gene Expression of Human Adipose–derived Mesenchymal Stem Cells: An in vitro Study

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