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. Author manuscript; available in PMC: 2020 Dec 14.
Published in final edited form as: J Orthop Res. 2018 Jul 13;36(11):2901–2910. doi: 10.1002/jor.24061

Chondrocyte and Mesenchymal Stem Cell Derived Engineered Cartilage Exhibits Differential Sensitivity to Pro-Inflammatory Cytokines

Bhavana Mohanraj 1,2, Alice H Huang 1,2, Meira J Yeger-McKeever 1, Megan J Schmidt 1, George R Dodge 1,2,3, Robert L Mauck 1,2,3,*
PMCID: PMC7735382  NIHMSID: NIHMS1612958  PMID: 29809295

Abstract

Tissue engineering is a promising approach for the repair of articular cartilage defects, with engineered constructs emerging that match native tissue properties. However, the inflammatory environment of the damaged joint might compromise outcomes, and this may be impacted by the choice of cell source in terms of their ability to operate anabolically in an inflamed environment. Here, we compared the response of engineered cartilage derived from native chondrocytes and mesenchymal stem cells (MSCs) to challenge by TNFα and IL-1β in order to determine if either cell type possessed an inherent advantage. Compositional (extracellular matrix) and functional (mechanical) characteristics, as well as the release of catabolic mediators (matrix metalloproteinases (MMPs), nitric oxide (NO)) were assessed to determine cell- and tissue- level changes following exposure to IL-1β or TNF-α. Results demonstrated that MSC-derived constructs were more sensitive to inflammatory mediators than chondrocyte-derived constructs, exhibiting a greater loss of proteoglycans and functional properties at lower cytokine concentrations. While MSCs and chondrocytes both have the capacity to form functional engineered cartilage in vitro, this study suggests that the presence of an inflammatory environment is more likely to impair the in vivo success of MSC-derived cartilage repair.

Introduction

Osteoarthritis (OA) is a progressive, degenerative joint disease characterized by articular cartilage fibrillation and erosion, leading to the loss of load-bearing function [1, 2]. Likewise, focal injuries to the cartilage surface do not heal, and can progress to larger sizes and instigate inflammation in the joint. Tissue engineering aims to address these issues via the combination of biomaterials, cells, and exogenous cues (e.g. growth factors, mechanical stimulation) to fabricate cartilage analogs for their in vivo application. Decades of work have culminated in the ability to engineer cartilage tissue that recapitulates the native tissue phenotype and structure-function properties [3–7]. Despite this achievement, these tissues are typically realized in vitro, under ‘optimal’ growth conditions, which are not representative of the implantation milieu. Indeed, inflammatory mediators in the injured and OA joint environment [1] will challenge the survival and growth of these constructs when they are implanted in vivo. Interleukin-1 (IL-1α and IL-1β) and tumor necrosis factor-alpha (TNF-α) are the primary cytokines that induce matrix catabolism and are detected at elevated levels in cartilage, synovial fluid, and synovium of OA joints [8–11]. While TNF-α is considered an early marker of OA, IL-1β is present at both early and late stages of degeneration [12]. In human OA cartilage, both cytokines co-localize with the expression and activity of matrix metalloproteinases (MMPs), which mediate degradation of the collagen network [13].

To investigate the mechanisms by which IL-1β and TNF-α induce cartilage degradation, a number of in vitro explant and engineered cartilage models have been developed. Treatment of newborn and adult bovine explants with IL-1β and TNF-α results in a dose-dependent increase in proteoglycan release, nitric oxide production, MMP synthesis, and cell death, as well as a decrease in collagen content and tissue mechanical properties [14–17]. In engineered cartilage fabricated using articular chondrocytes, sensitivity to cytokine-mediate changes has been evaluated as a function of construct maturity and in a biomaterial system. Constructs allowed to mature prior to IL-1β (or IL-1α) exposure were less susceptible to matrix degradation and showed attenuated loss of mechanical properties compared to those exposed at earlier time points [18–20]. Kwon et al. showed that biomaterial selection can also influence chondrocyte response to inflammation, where silk-based scaffolds appeared to be chondro-protective compared to PLA-based systems [21]. In addition, it has been noted that the ability of chondrocytes to robustly produce matrix and integrate with native cartilage is limited in an inflammatory environment [22].

While chondrocytes synthesize cartilage matrix components as an inherent function of their differentiated phenotype, autologous cells are of limited supply for regenerative medicine applications, particularly in patients with advanced OA. Mesenchymal stem cells (MSCs) are an alternative cell source that may be appropriate for cartilage repair. Bone marrow-derived MSCs can differentiate along chondrogenic, osteogenic, and adipogenic lineages when cultured in the appropriate scaffold environment and with defined chemical stimuli [23]. A number of studies have described MSC chondrogenesis in the presence of TGF-β to induce the production of proteoglycans and collagen type II, and some have reported the gradual development of near-native functional properties [24–28]. In addition, MSCs may also exhibit immunomodulatory functions [29, 30]. Despite the potential of this cell source, MSCs typically produce a less functional matrix compared to chondrocytes [31, 32], have a genome-wide expression profile that is distinct from chondrocytes even after differentiation [33], and are more susceptible to nutrient deprivation-induced cell death [34]. Based on these findings, it is likely that the response of MSC-based engineered cartilage will be distinct from that of chondrocyte-based engineered cartilage with cytokine challenge. Data supports that MSC-based engineered cartilage, similar to chondrocyte-based constructs, is affected by the presence of inflammatory cytokines. IL-1β and TNF-α inhibited MSC chondrogenesis in pellet and 3D constructs by down-regulating SOX9, COMP, collagen type II, and aggrecan gene expression, despite the continued presence of chondrogenic growth factors [35, 36]. These results suggest that MSCs are sensitive to cytokines during and after chondrogenesis.

To determine how cell source impacts the response of engineered cartilage to inflammatory stimuli, this study directly compared the effect of IL-1β and TNF-α on chondrocyte- and MSC-based constructs. Cellular- and tissue-level response to cytokine exposure was evaluated by measuring release of catabolic mediators, including nitric oxide and active MMP, as well as changes in matrix composition and mechanical properties. Findings from this study may have implications for the clinical use of chondrocyte- or MSC-derived engineered cartilage in the inflammatory environment of a damaged or osteoarthritic joint.

Methods

Engineered Cartilage Fabrication and Culture with Inflammatory Cytokines

Articular cartilage was harvested from the trochlear groove and femoral condyles of juvenile bovine knees (aged 2 to 6 months; Research 87, MA). Chondrocytes were isolated as previously described [37, 38]. Briefly, cartilage was minced, digested in collagenase for 18 to 24 hours at 37°C (type 2 collagenase, 298 U/mg, Worthington Biochemical Corp, NJ). The cell suspension was filtered through a 70 μm cell strainer and washed (2% penicillin/ streptomycin/ fungizone (PSF) in phosphate buffered saline, 1750 rpm for 15 minutes) to collect chondrocytes. Mesenchymal stem cells (MSCs) were isolated from bone marrow harvested from the tibia and femur of juvenile bovine knees and expanded in monolayer (P2 or P3), representing approximately 2 population doublings per passage after the cells reached initial confluence at passage 0 [31]. Each cell type was suspended in chemically defined medium (CM-, 40 million cells/mL) and combined with 4% w/v agarose (Type VII in PBS, Sigma, MO) in equal volumes to form a cell-agarose solution at a final cell concentration of 20 million cells/mL in 2% w/v agarose. The cell slurry was cast between glass plates, gelled at room temperature for 10 minutes, and biopsy punched to form uniform cylindrical constructs (Ø: 4mm, H: 2.25mm). Constructs were pre-cultured for 21 days in chemically defined medium containing 10ng/mL TGF-β (CM+, DMEM, 1% PSF, 1% ITS+ premix, 40 μg/mL L-proline, 50 μg/mL ascorbic acid, 0.1 μM dexamethasone, 0.5% v/v bovine serum albumin, and 100μg/mL sodium pyruvate [39]). Following this pre-culture period, chondrocyte- and MSC-seeded constructs were transferred to CM- (medium lacking TGF-β) and exposed to either IL-1β or TNF-α for 6 days. To evaluate dose-response, cytokines were added to the medium at increasing concentrations (1, 5, and 10 ng/mL). Medium and cytokines were refreshed on day 3, with medium harvested on days 3 and 6.

Assays to Measure Engineered Cartilage Response to Inflammatory Cytokines

Engineered cartilage mechanical properties were evaluated by unconfined compression testing (N=4 biological replicates per condition) after 6 days of cytokine exposure, as previously described [31]. Constructs were subjected to a 2 gram creep load, followed by a stress relaxation protocol (10% compressive strain applied at 0.05 strain/s held for 1000s) and dynamic loading (1% strain, 1Hz, 10 cycles) to measure the equilibrium and dynamic modulus [40]. For biochemical assays, constructs were subsequently lyophilized and papain digested overnight. Glycosaminoglycan (GAG) content was measured using the dimethylmethylene blue (DMMB) assay [41] and collagen content via the OHP assay [42]. GAG and OHP were normalized by construct wet weight. Medium (N=2 biological replicates per condition, for 5 constructs cultured together) was analyzed for GAG release, as well as established catabolic mediators, nitric oxide (NO, Griess assay, Promega, WI [43]) and MMP activity after APMA activation (Generic MMP 520 Fluorescence kit, Anaspec, CA). At the terminal time point, constructs were also fixed (4% paraformaldehyde), ethanol dehydrated, and paraffin embedded (N=1 per condition). Constructs were sectioned across the cross-sectional face (8μm sections) and stained with Alcian Blue to qualitatively assess changes in proteoglycan content. Immunohistochemistry (Universal Elite ABC Kit, R.T.U. VectaStain Kit, Vector Laboratories, CA) was also conducted to determine the spatial distribution of chondroitin sulfate (10μg/mL, II-II63B, DSHB, IA) and type II collagen (10μg/mL, 9BA12, DSHB, IA).

Statistical Analysis

The dose-dependent effects of IL-1β and TNF-α on mechanical properties and matrix content were analyzed using a one-way ANOVA with Tukey’s post-hoc test (p<0.05) for each cell type. GAG release, and NO and MMP production were evaluated using a two-way ANOVA with Tukey’s post-hoc test (p<0.05) for chondrocytes and MSCs individually, with time in culture (day 3 or day 6) and dose serving as independent variables. Difference in the magnitude of change between chondrocyte and MSC response, for each cytokine concentration, was also assessed using a two-way ANOVA with Bonferroni’s post-hoc test (p<0.05). Equilibrium modulus data was also fit to a single-phase exponential decay curve in order to test the hypothesis that chondrocyte and MSC responses could be fit with the same parameters. Rejection of this hypothesis would indicate differences in the rate of decay in mechanical properties as a function of cytokine concentration between the two cell types. Results shown are representative of two (chondrocytes) or three (MSCs) independent experiments. All statistical analysis was conducted using GraphPad Prism 6 (CA) and SYSTAT 13 (CA).

Results

Effect of Cytokines on Mechanical Properties and Matrix Composition

Exposure to either IL-1β or TNF-α significantly reduced mechanical properties in a dose-dependent manner in chondrocyte- and MSC-derived constructs. Although exposure to IL-1β or TNF-α at 1ng/mL induced a moderate decrease in equilibrium (Figure 1, A–B) and dynamic (Figure 1, C–D) moduli, a greater reduction in properties was observed at both 5 and 10ng/mL (no differences were found between these two groups). MSCs were more sensitive than chondrocytes to cytokine exposure for a given dose (e.g. EY, 5ng/mL, IL-1β: −79% CH, −97% MSCs, TNF-α: − 71% CH, −99% MSCs vs. control), with nearly complete loss of mechanical integrity at the highest concentrations of IL-1β and TNF-α assayed. A single-phase exponential decay curve fit to equilibrium modulus data showed that, for both cytokines, there was a significant increase in the decay rate constant (K) for MSCs compared to chondrocytes (Figure 1, E–F). ‘Half-life’ was also calculated for this response; that is, the cytokine concentration at which a 50% reduction in mechanical properties would be expected. For both IL-1β and TNF-α, chondrocytes required higher cytokine concentrations in order to elicit the same decrease as was observed for MSC-derived engineered cartilage (IL-1β, CH: 0.58 vs MSCs: 0.33 ng/mL and TNF-α, CH: 1.83 vs. MSCs 1.18 ng/mL).

Figure 1. Mechanical properties of MSC-derived engineered cartilage are more sensitive to cytokine challenge compared to chondrocyte-derived constructs.

Figure 1.

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Construct equilibrium (EY) and dynamic (G*) modulus show a dose-dependent effect for (A-C) IL-1β and (B-D) TNF-α. Cytokine challenge of MSC-derived cartilage shows a near complete loss of mechanical integrity at the highest cytokine concentrations (5 and 10ng/mL). (E-F) Fit of an exponential decay curve indicated an accelerated rate of decline in mechanical properties (K) for MSC- compared to chondrocyte-derived constructs. p<0.05 for * vs. control, + vs. 1ng/mL and control, S vs. MSC-derived constructs (based on fold change values) at the same cytokine concentration and time point.

GAG content in chondrocyte- and MSC-derived constructs was consistent with the dose-dependent effect of cytokines on mechanical properties (Figure 2, A–B). That is, MSC-derived constructs showed a greater magnitude of GAG loss compared to chondrocytes across all IL-1β or TNF-α concentrations (e.g. GAG %WW, 5ng/mL, IL-1β: −47% CH, −64% MSCs, TNF-α: −37% CH, −74% MSCs vs. control). In contrast, collagen content was relatively stable with cytokine challenge (Figure 2, C–D). Chondrocyte-derived constructs exposed to either IL-1β or TNF-α consistently showed a modest decrease in collagen content at concentrations of 5 and 10ng/mL. No significant differences in collagen content were observed in MSC-derived constructs in response to either cytokine. Alcian blue staining and immunohistochemistry (IHC) for proteoglycans and chondroitin sulfate, respectively, visually confirmed the quantitative changes in GAG content (Figure 3 and 4). Compared to chondrocyte-derived constructs, MSC-derived constructs showed a marked loss in staining intensity for proteoglycans with the greatest changes seen at the highest concentrations of IL-1β and TNF-α. In contrast, IHC for type II collagen showed little change with exposure to either cytokine, except at the highest concentration of TNF-α.

Figure 2. Matrix loss from engineered constructs with exposure to IL-1β or TNF-α.

Figure 2.

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(A-B) GAG content in MSC-derived constructs decreased to a greater extent than chondrocyte-derived constructs, and in a dose-dependent manner. Conversely, collagen content (C-D) was largely unaffected by cytokine exposure. Chondrocyte-derived constructs showed a moderate decrease in response to IL-1β or TNF-α exposure; no significant differences were found for MSCs. p<0.05 for * vs. control, + vs. 1ng/mL and control, S vs. MSC-derived constructs (based on fold change values) at the same cytokine concentration and time point.

Figure 3. Histological assessment of engineered constructs confirmed biochemical measurements following exposure to IL-1β.

Figure 3.

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Compared to (A) chondrocyte-derived constructs, IL-1β induced a greater progressive loss of staining intensity for proteoglycans (Alcian blue) and chondroitin sulfate (CS) in (B) MSC-derived constructs, most notably at 5 and 10ng/mL. Collagen Type II staining was not markedly affected by cytokine exposure.

Figure 4. Histological assessment of engineered constructs confirmed biochemical measurements following exposure to TNF-α.

Figure 4.

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Staining for proteoglycans (Alcian blue) and chondroitin sulfate (CS) was modestly reduced in (A) chondrocyte-derived constructs as compared to the marked decrease observed in (B) MSC-derived constructs. Collagen Type II staining was minimally affected by cytokine exposure, with changes only observed at the highest cytokine concentration.

Cytokine-mediated Release of GAG, NO and MMPs

Catabolic mediators transiently increased in response to cytokine challenge, as measured by nitric oxide release to the medium and detection of MMPs. Nitric oxide production (measured as nitrite, a byproduct of NO breakdown) was dose-dependent and was consistently greater on day 3 compared to day 6, and for chondrocyte- compared to MSC-derived constructs at higher concentrations (Figure 5, A–B). In comparison, the pattern of MMP production was both cytokine-specific and cell type-dependent. Although absolute MMP levels between chondrocyte- and MSC-derived constructs was similar, differences relative to un-treated controls were observed with IL-1β treatment. Chondrocyte-derived constructs showed an increase in relative production compared to MSC-derived cartilage on both day 3 at 10ng/mL (CH: 27.6x CH, MSCs: 3.5x vs. control) and day 6 (CH: 26.2x, MSCs: 6x vs. control) at the highest concentrations (Figure 5C). However, MSC-derived constructs were sensitive to IL-1β only at the lowest concentration (1ng/mL) on day 6 and showed an increase in overall levels compared to day 3. A markedly different response was observed in constructs following exposure to TNF-α (Figure 5D). By day 6, activated MMP levels were significantly increased for MSC-derived constructs compared to control and chondrocytes for all TNF-α concentrations (e.g. 10ng/mL, CH: 0.6x, MSCs: 6.4x vs. control).

Figure 5. Differential release of catabolic mediators and matrix components by chondrocyte- and MSC-derived engineered cartilage in response to cytokine challenge.

Figure 5.

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(A-B) Nitric oxide production (nitrite) was higher for chondrocyte-derived constructs compared to MSCs, and on day 3 compared to day 6, for both cytokines and across all doses. (C-D) In response to IL-1β, MMP activity increased from day 3 to day 6, in a dose-dependent manner. Although absolute measurement of activated MMPs was higher for MSC-derived constructs, the relative increase compared to control was larger for chondrocytes due to a lower baseline level. In contrast, TNF-α exposure resulted in only low levels of MMP activity on day 3 for both chondrocytes and MSCs; however, by day 6 MMP activity for MSC-derived constructs increased markedly and matched that resulting from IL-1β exposure. (E-F) GAG release to the media was consistently lower for chondrocyte-derived constructs than MSCs on day 3 for both cytokines. However, by day 6, minimal differences in matrix loss were observed between both cell types. p<0.05 for * vs. control, + vs. 1ng/mL and control, ∂ vs. 5ng/mL, 1ng/mL, and control, # vs. day 6 for the same cell type, S vs. MSC-derived constructs (based on fold change values for activated MMP and released GAG measurements) at the same cytokine concentration and time point.

To determine how these catabolic mediators effect matrix degradation, GAG release to the medium was also measured (Figure 5, E–F). In response to either cytokine, GAG release from chondrocyte-derived constructs was lower on day 3 compared to day 6, at concentrations of 5 and 10ng/mL. No differences were observed between time points for MSC-derived constructs, with GAG release maintained at an elevated level throughout the culture period. For IL-1β, matrix release was also higher for MSC- compared to chondrocyte-derived constructs on day 3 (e.g. 10ng/mL, CH: 1.9x, MSCs: 3.3x vs. control), but by day 6 the chondrocyte response was equivalent (e.g. 10ng/mL, CH: 5.5x, MSCs: 4x vs. control) (Figure 5E). Following exposure to TNF-α, there was a difference in release between cell types on day 3 (e.g. 10ng/mL, CH: 1.9x, MSCs: 4.2x vs. control). However, by day 6 GAG release from chondrocyte-derived constructs matched that of MSCs (e.g. 10ng/mL, CH: 4x, MSCs: 5x vs. control; Figure 5F).

Discussion

The inflammatory environment of an injured or osteoarthritic joint presents a significant challenge for the success of tissue engineered cartilage strategies. Here, we show that the inflammatory cytokines, IL-1β and TNF-α, stimulate the release of catabolic mediators (including NO and MMPs) that likely initiate matrix loss, which directly impacts construct functional properties. These factors, in combination with cytokine-mediated inhibition of matrix biosynthesis, may prevent cell-based constructs from effectively restoring load-bearing capacity in a diseased environment. Two of the primary cell sources utilized for cartilage tissue engineering are native chondrocytes and mesenchymal stem cells. Although both cell types have been successfully used to engineer cartilage in vitro [3–7], challenges remain in repairing cartilage within the in vivo joint environment [44–49]. To determine whether inflammation-induced degeneration is dependent on cell source in this context, we directly compared the response of chondrocyte- and MSC-derived constructs to IL-1β and TNF-α in an in vitro model system. While both cell types showed a dose-dependent reduction in mechanical properties, MSC-derived constructs were more sensitive to inflammatory challenge, with complete loss of structural integrity measured at higher cytokine concentrations. Consistent with these observations, construct GAG content and histological staining intensity were markedly reduced; however, collagen content was only minimally affected by cytokine exposure. Although collagen contributes to both equilibrium and dynamic mechanical properties [50, 51], the loss of GAG may play a larger role in engineered cartilage due to the immaturity of the collagen network (<1%WW vs. 15–20%WW in native tissue [52]).

Our results complement previous studies describing matrix depletion in response to IL-1β and TNF-α in engineered tissues using chondrocytes [18–20, 53] and MSCs [35, 54], as well as iPSCs [55]. Similarities across cell types include ‘maturation-dependent’ effects of cytokines that have previously been characterized for chondrocytes constructs [18–20], and more recently for MSCs. Ousema et al. showed that IL-α significantly impaired MSC chondrogenesis in woven PCL scaffolds when present in the media from the onset of culture (with TGF-β3) as compared to constructs pre-cultured for 2 weeks prior to cytokine treatment [54]. This inhibition of chondrogenic (and osteogenic) differentiation of MSCs (29, 30, 47) may be due to inhibition of Sox-9 transcriptional activity, which is critical for collagen and aggrecan synthesis [56].

To determine if cell source affects the production of catabolic factors, nitric oxide and MMPs were measured in culture medium during exposure of constructs to IL-1β and TNF-α. NO promotes cartilage degradation through inhibition of aggrecan and collagen synthesis, activation of MMPs, and increased susceptibility to other oxidants that cause apoptosis [57]. Activated MMPs further contribute to destruction of the collagen network [13, 58] and, together with NO, may mediate the loss of construct functional properties and biochemical composition seen in this work. Higher NO concentrations were consistently found in the media of chondrocyte-derived constructs cultured with either IL-1β or TNF-α, as compared to MSCs at both time points (days 3 and 6). In comparison, MMP production showed a differential response that depended on both the cytokine and cell type. Although the absolute measurements of MMP levels were higher for MSC-derived constructs than chondrocytes following IL-1β treatment, the baseline level of activity in naïve constructs was also higher. As a result, the relative increase in MMP production (vs. control) was greater for chondrocyte-derived constructs than MSCs, though these findings suggest that MSC-derived constructs are degrading formed matrix at a higher rate in naïve (control) constructs. Despite the overall increase in the activity of catabolic mediators for chondrocyte-derived constructs, GAG released to the media was initially higher for MSC-derived constructs on day 3.

The exacerbated matrix loss from MSC-derived constructs supported the observed changes in mechanical and biochemical properties, and suggests that MSCs may exhibit greater sensitivity to IL-1β than chondrocytes. These results contrast with the response to TNF-α treatment, where only MSC-derived constructs showed a marked increase in activated MMPs on day 6 of culture. Although NO production was elevated for chondrocytes throughout, and MMP levels were only elevated at the later time point for MSCs, GAG released to the media was still higher for MSC-derived constructs than chondrocytes on day 3. Similar to the effect of IL-1β, MSCs seem to be more sensitive to TNF-α than chondrocytes in the context of engineered cartilage. Notably, the temporal patterns of NO and MMP production were different for each cytokine and cell type, highlighting the need for additional studies to investigate the regulation of relevant pro-inflammatory signaling pathways. The NF-κB pathway is of particular interest, since IL-1β and TNF-α induce catabolic activity through associated signaling cascades [35, 59]. This pathway is a primary target for therapeutic intervention in OA, and studies have focused on the development of in vitro, engineered cartilage models to investigate the effect of NF-κB inhibition (e.g., small molecules (SC-514 [55]) and dominant negative expression of IκB [35]). Recent work by Brunger et al. also demonstrated that genome engineering (CRISPR/Cas9) can be used to create stem cells that produce antagonists in response to IL-1 and TNF-α treatment in an auto-regulated manner to protect against cartilage degradation [60]. In this context, the choice of cell source for cartilage tissue engineering may not only influence the likelihood of repair in vivo, but also the response to candidate therapeutics for the treatment of OA.

Overall, this study demonstrated that cell source (native chondrocytes vs. chondrogenically differentiated MSCs) influences the response of engineered cartilage to pro-inflammatory cytokines. MSC-derived constructs were more responsive than chondrocytes to IL-1β and TNF-α, with greater loss of matrix and functional properties at lower doses of cytokine challenge. While elevated levels of NO and MMPs were generally observed for chondrocyte-derived constructs, exacerbated construct degeneration was observed for MSCs, indicating an increased sensitivity of MSCs to catabolic mediators. Although MSCs and chondrocytes both have the capacity to produce matrix in engineered constructs, fundamental differences exist between the two cell types that impact their potential regenerative capacity upon in vivo implantation. Our findings illustrate these differences in the context of a cytokine challenge, and further support the notion that choice of cell source in tissue engineering will influence the likelihood of successful repair within the inflammatory environment of OA.

Supplementary Material

Supplemental Table 1

Acknowledgements

This work was supported by the Department of Veterans Affairs (I01 RX001213), the National Institutes of Health (R01 EB008722), and a National Science Foundation Research Experience for Undergraduates (REU) program (SUNFEST, EEC 1359107). The authors have no conflicts of interest.

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Supplementary Materials

Supplemental Table 1
NIHMS1612958-supplement-Supplemental_Table_1.docx (16.2KB, docx)

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