Non-linear optical microscopy as a novel quantitative and label-free imaging modality to improve the assessment of tissue-engineered cartilage

Islam, Ashraful; Romijn, Elisabeth I.; Lilledahl, Magnus B.; Martínez-Zubiaurre, Iñigo

doi:10.1016/j.joca.2017.06.008

Cited by 8 publications

(5 citation statements)

References 34 publications

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“…Macroscopically-normal-looking cartilage, corresponding to cartilage areas looking intact to the eye and presenting no signs of abnormalities, such as tears, scratches, rugose or eroded tissue, was collected from femoral condyles during total knee replacements and was used to isolate human chondrocytes. All cell types were isolated using a mixed enzymatic-explant method, as previously described (Islam et al, 2016;Islam et al, 2017). Briefly, all tissue specimens were washed three times with sterile Dulbecco's phosphate buffered saline (PBS; D8537, Sigma-Aldrich) and minced into small pieces for subsequent enzymatic digestion in collagenase XI solution (≥ 800 units/mg solid; C9407, Sigma-Aldrich) at a final concentration of 1.25 mg/mL at 37 °C on a shaker at 400 rpm.…”

Section: Isolation and Culture Of Human Stromal Cellsmentioning

confidence: 99%

“…Liechty et al (2000) propose that MSCs promote tissue regeneration by engraftment of cells in damaged areas and transdifferentiation into tissue-forming cells to promote repair. Recently, paracrine signalling and release of potent bioactive factors have emerged as modulators of the microenvironment during tissue healing (Gnecchi et al, 2016;Iso et al, 2007;Prockop, 2009). In the field of cartilage repair and OA, the fate of implanted cells during biological repair procedures and their contribution to rebuilding the damaged tissue is mostly unknown.…”

Section: Introductionmentioning

confidence: 99%

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Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells

Islam¹,

Urbárová²,

Bruun³

et al. 2019

eCM

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Mesenchymal stromal cells (MSCs), given their regenerative potential, are being investigated as a potential therapeutic tool for cartilage lesions. MSCs express several bioactive molecules which act in a paracrine fashion to modulate the tissue microenvironment. Yet, little is known about the divergence of these signalling molecules in different MSC populations. The present study investigated secretomes of stromal cells harvested from Hoffa's fat pad (HFPSCs), synovial membrane (SMSCs), umbilical cord (UCSCs) and cartilage (ACs) by quantitative liquid chromatography-mass spectrometry (LC-MS/MS) proteomics. Also, multiplex protein arrays and functional assays were performed to compare the constitutive immunomodulatory capabilities of different MSCs. Proteins involved in extracellular matrix degradation and inflammation, such as matrix metalloproteinases (MMPs), interleukin (IL)-17 and complement factors, were downregulated in UCSCs as compared to adult cell sources. Additionally, secretion of transforming growth factor (TGF)-β1 and prostaglandin E2 (PGE2) was enhanced in UCSC supernatants. UCSCs were superior in inhibiting peripheral blood mononuclear cell (PBMC) proliferation, migration and cytokine secretion as compared to adult stromal cells. SMSCs significantly suppressed the proliferation of PBMCs only if they were primed with pro-inflammatory cytokines. Although all cell types repressed human leukocyte antigen-DR isotype (HLA-DR) surface expression and cytokine release by activated macrophages, only UCSCs significantly blocked IL-6 and IL-12 production. Furthermore, UCSCs supernatants increased aggrecan gene expression in twodimensional chondrocyte cultures. The data demonstrated that UCSCs displayed superior anti-inflammatory and immunosuppressive properties than stromal cells from adult tissues. This allogeneic cell source could potentially be considered as an adjuvant therapy for articular cartilage repair.

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Section: Isolation and Culture Of Human Stromal Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells

Islam¹,

Urbárová²,

Bruun³

et al. 2019

eCM

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“…The feasibility of SHM for the assessment of articular cartilage in animals [11,28,30,56], humans [12,33], and tissue-engineered [44,48,57,58] cartilage constructs have been demonstrated several times. SHM can provide a wealth of information at several levels, including, but not limited to, micron-level surface morphology, changes in collagen architecture, detection of collagen types in regenerated cartilage, label-free analysis, and 3D imaging capability (optical biopsy) within the limit of laser penetration depth.…”

Section: Discussionmentioning

confidence: 99%

“…An initial effort has been undertaken by Hayley et al, which demonstrated that cartilage damage is associated with collagen redistribution, and collagen disorganization could be assigned an independent score [10]. A similar effort associated with SHM scoring system for the assessment of tissue-engineered cartilage has been also reported by Islam et al [48].…”

Section: Challenges To Shm For Cartilage Assessmentmentioning

confidence: 95%

Assessment of Articular Cartilage by Second Harmonic Microscopy: Challenges and Opportunities

Kumar

2019

Front. Phys.

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Second harmonic microscopy (SHM) has emerged as a powerful non-destructive optical imaging modality that has high potential to perform the advanced structural characterization of intact articular cartilage. This article provides the current status, opportunities, and challenges of SHM for the use in cartilage evaluation. A novel perspective on SHM is being addressed that includes the reason, need and the importance of SHM in the assessment of articular cartilage. By taking the advantage of collagen specificity and high-resolution imaging, SHM can detect the structural and morphological changes in early stage cartilage damage that would otherwise be overlooked by standard clinical imaging methods such as radiography and arthroscopy. The current status of SHM for articular cartilage analysis and the major progress reported in recent literature are briefly described herein. We summarize the status of scientific efforts and the challenges that need to overcome before moving toward clinical applications of SHM in the context of cartilage evaluation.

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“…Sections were imaged by bright field light microscopy (Leica DMI6000B). To quantify the in vitro chondrogenic potential, a visual semi-quantitative scoring of tissue sections (Bern score) was applied independently by three different observers [26]. The chondrogenic potential was classified into two groups according to histological outcomes: “Group A” with high chondrogenic potential (Bern score 6–9) and “Group B” with low chondrogenic potential (Bern score < 6) (Table 1).…”

Section: Methodsmentioning

confidence: 99%

In vitro chondrogenic potency of surplus chondrocytes from autologous transplantation procedures does not predict short-term clinical outcomes

Islam

Fossum

Hansen

et al. 2019

BMC Musculoskelet Disord

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BackgroundAutologous chondrocyte implantation (ACI) has been used over the last two decades to treat focal cartilage lesions aiming to delay or prevent the onset of osteoarthritis; however, some patients do not respond adequately to the procedure. A number of biomarkers that can forecast the clinical potency of the cells have been proposed, but evidence for the relationship between in vitro chondrogenic potential and clinical outcomes is missing. In this study, we explored if the ability of cells to make cartilage in vitro correlates with ACI clinical outcomes. Additionally, we evaluated previously proposed chondrogenic biomarkers and searched for new biomarkers in the chondrocyte proteome capable of predicting clinical success or failure after ACI.MethodsThe chondrogenic capacity of chondrocytes derived from 14 different donors was defined based on proteoglycans staining and visual histological grading of tissues generated using the pellet culture system. A Lysholm score of 65 two years post-ACI was used as a cut-off to categorise “success” and “failure” clinical groups. A set of predefined biomarkers were investigated in the chondrogenic and clinical outcomes groups using flow cytometry and qPCR. High-throughput proteomics of cell lysates was used to search for putative biomarkers to predict chondrogenesis and clinical outcomes.ResultsVisual histological grading of pellets categorised donors into “high” and “low” chondrogenic groups. Direct comparison between donor-matched in vitro chondrogenic potential and clinical outcomes revealed no significant associations. Comparative analyses of selected biomarkers revealed that expression of CD106 and TGF-β-receptor-3 was enhanced in the low chondrogenic group, while expression of integrin-α1 and integrin-β1 was significantly upregulated in the high chondrogenic group. Additionally, increased surface expression of CD166 was observed in the clinical success group, while the gene expression of cartilage oligomeric matrix protein was downregulated. High throughput proteomics revealed no differentially expressed proteins from success and failure clinical groups, whereas seven proteins including prolyl-4-hydroxylase 1 were differentially expressed when comparing chondrogenic groups.ConclusionIn our limited material, we found no correlation between in vitro cartilage-forming capacity and clinical outcomes, and argue on the limitations of using the chondrogenic potential of cells or markers for chondrogenesis as predictors of clinical outcomes.Electronic supplementary materialThe online version of this article (10.1186/s12891-018-2380-4) contains supplementary material, which is available to authorized users.

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Non-linear optical microscopy as a novel quantitative and label-free imaging modality to improve the assessment of tissue-engineered cartilage

Abstract: SHG represents a reliable quantitative approach to assess collagen deposition in laboratory engineered cartilage, and may be applied to improve currently established scoring systems.

Cited by 8 publications

References 34 publications

Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells

Large-scale secretome analyses unveil the superior immunosuppressive phenotype of umbilical cord stromal cells as compared to other adult mesenchymal stromal cells

Assessment of Articular Cartilage by Second Harmonic Microscopy: Challenges and Opportunities

In vitro chondrogenic potency of surplus chondrocytes from autologous transplantation procedures does not predict short-term clinical outcomes

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