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Isolation and characterization of synovial fluid mesenchymal stem cells from patients with and without knee osteoarthritis
S406
Abstracts / Osteoarthritis and Cartilage 25 (2017) S76eS444
ing fibrosis, and elevated IL6 potentially driving cartilage catabolism and being associated with pain. The majority of the changes induced by Hymovis in the current study reflect beneficial/disease-modifying effects - decreasing fibrosis by reducing fibronectin and collagen synthesis and increasing turnover by lowering the TIMP/MMP ratio, reducing potential ADAMTS5-driven aggrecanolysis, and reducing synthesis of the proinflammatory and pro-algesic cytokine IL-6. In all cases, the effect of Hymovis was similar in the presence of Wnt3a, thus reversing the potentially negative impact of this molecule in the OA joint. It is unclear what drives the differential response of synovial fibroblasts from various patients, potentially reflecting differences in OA phenotype (e.g. degree of synovitis), and/or individual genetic differences between patients. 668 ISOLATION AND CHARACTERIZATION OF SYNOVIAL FLUID MESENCHYMAL STEM CELLS FROM PATIENTS WITH AND WITHOUT KNEE OSTEOARTHRITIS E.B. de Sousa y, z, R.S. da Costa y, M.L. Duarte y, V.M. Neto z, x, D.P. Aguiar y. y Natl. Inst. of Traumatology and Orthopedics, Rio de Janeiro, Brazil; z Inst. of BioMed. Sci., Federal Univ. of Rio de Janeiro, Rio de Janeiro, Brazil; x Brain State Inst. Paulo Niemeyer, Rio de Janeiro, Brazil Purpose: Characterize the cells of the synovial fluid (SF) in patients with and without osteoarthritis (OA) according to the International Society of Cell Therapy. Methods: Synovial fluid was collected from patients submitted to knee arthroscopy (n ¼ 23 K-L 0) or knee replacement (n ¼ 37, 9 K-L 3 and 28 K-L 4), and centrifuged to isolate cells from the fluid. Cells were cultured and identified with markers specific for MSC. Cultures were then exposed to specific cocktail to induce chondrogenic, osteogenic and adipogenic differentiation. Medium from induction protocols was submitted to ELISA assay in order to evaluate BMP4 concentration. Results: Samples from patients with OA presented higher number of CFU than the ones without OA (with OA 74,17 ± 1,07 vs. without OA 2,16 ± 0,36 CFU per dishes, p < 0,0001, n ¼ 12). Besides, the colony diameter was also higher in patients with OA (with OA 6,62 ± 0,28 mm, n ¼ 23, vs. without OA 1,51 ± 0,17 mm, p < 0,0001, n ¼ 12). After 21 days, the colony-forming cells from OA patients differentiated into the three mesenchymal cell lineages, after the appropriate induction protocols. t find any significantly differences when comparing groups Results didn in relation to percentage of cells positive for CD73 and CD90 (61,15 ± 11,35 % without OA versus 90,30 ± 6,46% with OA, p ¼ 0,09), CD73, CD90 and CD105 (0,15 ± 0,15% without OA versus 0,60 ± 0,45% with OA, p ¼ 0,50), CD73, CD90 and CD146 (3,60 ± 3,0% without OA versus 7,73 ± 6,09% with OA, p ¼ 0,64), and CD73, CD90, CD105 and CD146 (0,15 ± 0,15% without OA versus 0,36 ± 0,31% with OA, p ¼ 0,64). In samples from chondrogenic induction, BMP4 expression was higher in patients with OA group all over time when compared to patients without OA. In the osteogenic induction samples, BMP4 expression was higher in patients with OA group when compared to the samples from patients without OA only at the 7th and 21st days. However, in the adipogenic induction samples, BMP4 expression was higher in patients without OA all over time in comparison to patients with OA. BMP4 expression at day 21 in the samples from chondrogenic induction was the highest of all. Conclusions: SF contains a MSC population, which forms a higher number of colonies, also with higher diameter in patients with OA. Cells did not differ in relation to MSC markers and were not positive to haematopoietic markers. BMP4 expression is higher in SF MSC of patients with OA after three lineage induction. The highest levels of BMP4 were found after chondrogenic differentiation.
Therapy e Non-Pharmacologic 669 CLINICAL CHANGES USING LATERAL WEDGES WITH AND WITHOUT ARCH SUPPORT IN PEOPLE WITH MEDIAL KNEE OSTEOARTHRITIS AND FLAT FEET: A RANDOMIZED CROSSOVER STUDY M.A. Hunt y, J. Takacs y, N.M. Krowchuk y, G.L. Hatfield z, R.S. Hinman x, R. Chang k. y Univ. of British Columbia, Vancouver, BC, Canada; z Univ. of The Fraser Valley, Chilliwack, BC, Canada; x Univ. of Melbourne, Melbourne, Australia; k Reebok Intl., Boston, MA, USA Purpose: Shoe-worn insoles built up along the lateral border - termed lateral wedges - have received much attention in the knee osteoarthritis (OA) literature for their ability to redistribute dynamic knee joint loads. Specifically, they have been shown to reduce the external knee adduction moment (KAM) - an indirect measure of mediolateral tibial load distribution. However, there have been highly variable biomechanical outcomes amongst individuals, and little evidence to support clinical improvement questions their overall effectiveness. Further, use of lateral wedges in people with flat feet may be contraindicated due to their design and interaction with that specific foot anatomy. Instead, people with flat feet - regardless of the presence of knee OA - may be more likely to receive arch support insoles. Since foot pain is common in people with knee OA, it is important to assess the effect of shoe-worn insoles such as lateral wedges on not only knee symptoms, but foot symptoms as well. Therefore, the aim of this feasibility study was to compare changes in knee and foot symptoms in people with medial tibiofemoral knee osteoarthritis (OA) and concomitant flat feet with the use of lateral wedge insoles with and without arch support. Methods: Twenty-six people with medial compartment knee OA and flat feet (quantified as þ4 or greater on the Foot Posture Index scale) participated in an exploratory randomized crossover study comparing five degree lateral wedges with and without customized foot arch support. Each intervention was worn for two months, separated by a two-month washout period of no wear. Main outcomes included the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain and physical function subscales, the revised short-form Foot Function Index (FFI) pain and stiffness subscales, and the timed stair climb test. Adverse events, patient preference and self-reported foot comfort was also assessed. Regression modeling was conducted to examine treatment, period, and interaction effects for each outcome. Significance was assessed at an alpha level of 0.05. Results: Twenty-two participants completed the trial, and no carryover or interaction effects were observed for any outcome. Significant betweencondition effects were only observed for the timed stair climb test, with greater improvements seen with the lateral wedges with arch support. Within-condition significant improvements were observed for all main outcomes (WOMAC pain and physical function, FFI pain and stiffness, and timed stair climb) following lateral wedge with arch support use (Table 1). More adverse effects were reported with the lateral wedges alone, and more people preferred the lateral wedges with arch support overall (17 out of 22). Finally, though the lateral wedges with arch support were reported to be more comfortable (6.9 ± 2.5 out of 10 vs 6.4 ± 2.6 out of 10), the between-condition difference was not statistically significant. Conclusions: Our study showed that the addition of customized arch support to a standard lateral wedge insole may improve foot and knee symptoms in people with knee OA and concomitant flat feet. Importantly, it was found that these “supported lateral wedges” can be safely delivered to a subgroup of people with knee OA (those with flat feet) for whom lateral wedges may have previously not been advocated. These findings support addressing foot mechanics when designing shoe-worn insoles for treatment of knee OA.
Table 1 Outcome data. Mean (sd) baseline and follow-up outcome data for all variables, as well as model adjusted differences (follow-up minus baseline; beta coefficient (95% confidence intervals)) within each condition as well as between conditions. Model estimate values represent the within- and between-condition least-square mean differences adjusted for period and random subject effects from the regression modeling. Lateral Wedges
Lateral Wedges plus Arch Support
Between-conditions comparison
Variable
Baseline
Follow-up
Difference
Baseline
Follow-up
Difference
Difference
Stair climb (seconds) WOMAC Pain (0-20) WOMAC Function (0-68) FFI Pain(25-100%) FFI Stiffness (25-100%)
5.32 (1.68) 5.9 (3.0) 20.2 (11.0) 41.0 (17.4) 39.2 (15.7)
5.35 (1.54) 5.2 (2.9) 17.6 (8.9) 38.4 (11.9) 37.1 (13.7)
0.04 (0.55, 0.63) 0.7 (1.8, 0.5) 2.5 (6.2, 1.1) 2.7 (8.9, 3.5) 2.1 (7.3, 3.0)
6.57 (4.04) 6.3 (3.4) 21.4 (12.5) 43.6 (16.2) 42.8 (19.5)
5.58 (2.29) 4.0 (2.5) 14.0 (8.8) 33.0 (9.3) 32.3 (9.4)
0.69 (1.3, 0.08)* 1.9 (3.1, 0.6)* 6.1 (9.9, 2.3)* 7.0 (13.5,0.6)* 6.3 (11.7, 0.9)*
0.73 (1.36, 0.11)* 1.2 (2.9, 0.5) 3.6 (7.9, 0.8) 4.3 (12.0, 3.4) 4.1 (11.0, 2.7)
*p < 0.05
Abstracts / Osteoarthritis and Cartilage 25 (2017) S76eS444
ing fibrosis, and elevated IL6 potentially driving cartilage catabolism and being associated with pain. The majority of the changes induced by Hymovis in the current study reflect beneficial/disease-modifying effects - decreasing fibrosis by reducing fibronectin and collagen synthesis and increasing turnover by lowering the TIMP/MMP ratio, reducing potential ADAMTS5-driven aggrecanolysis, and reducing synthesis of the proinflammatory and pro-algesic cytokine IL-6. In all cases, the effect of Hymovis was similar in the presence of Wnt3a, thus reversing the potentially negative impact of this molecule in the OA joint. It is unclear what drives the differential response of synovial fibroblasts from various patients, potentially reflecting differences in OA phenotype (e.g. degree of synovitis), and/or individual genetic differences between patients. 668 ISOLATION AND CHARACTERIZATION OF SYNOVIAL FLUID MESENCHYMAL STEM CELLS FROM PATIENTS WITH AND WITHOUT KNEE OSTEOARTHRITIS E.B. de Sousa y, z, R.S. da Costa y, M.L. Duarte y, V.M. Neto z, x, D.P. Aguiar y. y Natl. Inst. of Traumatology and Orthopedics, Rio de Janeiro, Brazil; z Inst. of BioMed. Sci., Federal Univ. of Rio de Janeiro, Rio de Janeiro, Brazil; x Brain State Inst. Paulo Niemeyer, Rio de Janeiro, Brazil Purpose: Characterize the cells of the synovial fluid (SF) in patients with and without osteoarthritis (OA) according to the International Society of Cell Therapy. Methods: Synovial fluid was collected from patients submitted to knee arthroscopy (n ¼ 23 K-L 0) or knee replacement (n ¼ 37, 9 K-L 3 and 28 K-L 4), and centrifuged to isolate cells from the fluid. Cells were cultured and identified with markers specific for MSC. Cultures were then exposed to specific cocktail to induce chondrogenic, osteogenic and adipogenic differentiation. Medium from induction protocols was submitted to ELISA assay in order to evaluate BMP4 concentration. Results: Samples from patients with OA presented higher number of CFU than the ones without OA (with OA 74,17 ± 1,07 vs. without OA 2,16 ± 0,36 CFU per dishes, p < 0,0001, n ¼ 12). Besides, the colony diameter was also higher in patients with OA (with OA 6,62 ± 0,28 mm, n ¼ 23, vs. without OA 1,51 ± 0,17 mm, p < 0,0001, n ¼ 12). After 21 days, the colony-forming cells from OA patients differentiated into the three mesenchymal cell lineages, after the appropriate induction protocols. t find any significantly differences when comparing groups Results didn in relation to percentage of cells positive for CD73 and CD90 (61,15 ± 11,35 % without OA versus 90,30 ± 6,46% with OA, p ¼ 0,09), CD73, CD90 and CD105 (0,15 ± 0,15% without OA versus 0,60 ± 0,45% with OA, p ¼ 0,50), CD73, CD90 and CD146 (3,60 ± 3,0% without OA versus 7,73 ± 6,09% with OA, p ¼ 0,64), and CD73, CD90, CD105 and CD146 (0,15 ± 0,15% without OA versus 0,36 ± 0,31% with OA, p ¼ 0,64). In samples from chondrogenic induction, BMP4 expression was higher in patients with OA group all over time when compared to patients without OA. In the osteogenic induction samples, BMP4 expression was higher in patients with OA group when compared to the samples from patients without OA only at the 7th and 21st days. However, in the adipogenic induction samples, BMP4 expression was higher in patients without OA all over time in comparison to patients with OA. BMP4 expression at day 21 in the samples from chondrogenic induction was the highest of all. Conclusions: SF contains a MSC population, which forms a higher number of colonies, also with higher diameter in patients with OA. Cells did not differ in relation to MSC markers and were not positive to haematopoietic markers. BMP4 expression is higher in SF MSC of patients with OA after three lineage induction. The highest levels of BMP4 were found after chondrogenic differentiation.
Therapy e Non-Pharmacologic 669 CLINICAL CHANGES USING LATERAL WEDGES WITH AND WITHOUT ARCH SUPPORT IN PEOPLE WITH MEDIAL KNEE OSTEOARTHRITIS AND FLAT FEET: A RANDOMIZED CROSSOVER STUDY M.A. Hunt y, J. Takacs y, N.M. Krowchuk y, G.L. Hatfield z, R.S. Hinman x, R. Chang k. y Univ. of British Columbia, Vancouver, BC, Canada; z Univ. of The Fraser Valley, Chilliwack, BC, Canada; x Univ. of Melbourne, Melbourne, Australia; k Reebok Intl., Boston, MA, USA Purpose: Shoe-worn insoles built up along the lateral border - termed lateral wedges - have received much attention in the knee osteoarthritis (OA) literature for their ability to redistribute dynamic knee joint loads. Specifically, they have been shown to reduce the external knee adduction moment (KAM) - an indirect measure of mediolateral tibial load distribution. However, there have been highly variable biomechanical outcomes amongst individuals, and little evidence to support clinical improvement questions their overall effectiveness. Further, use of lateral wedges in people with flat feet may be contraindicated due to their design and interaction with that specific foot anatomy. Instead, people with flat feet - regardless of the presence of knee OA - may be more likely to receive arch support insoles. Since foot pain is common in people with knee OA, it is important to assess the effect of shoe-worn insoles such as lateral wedges on not only knee symptoms, but foot symptoms as well. Therefore, the aim of this feasibility study was to compare changes in knee and foot symptoms in people with medial tibiofemoral knee osteoarthritis (OA) and concomitant flat feet with the use of lateral wedge insoles with and without arch support. Methods: Twenty-six people with medial compartment knee OA and flat feet (quantified as þ4 or greater on the Foot Posture Index scale) participated in an exploratory randomized crossover study comparing five degree lateral wedges with and without customized foot arch support. Each intervention was worn for two months, separated by a two-month washout period of no wear. Main outcomes included the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain and physical function subscales, the revised short-form Foot Function Index (FFI) pain and stiffness subscales, and the timed stair climb test. Adverse events, patient preference and self-reported foot comfort was also assessed. Regression modeling was conducted to examine treatment, period, and interaction effects for each outcome. Significance was assessed at an alpha level of 0.05. Results: Twenty-two participants completed the trial, and no carryover or interaction effects were observed for any outcome. Significant betweencondition effects were only observed for the timed stair climb test, with greater improvements seen with the lateral wedges with arch support. Within-condition significant improvements were observed for all main outcomes (WOMAC pain and physical function, FFI pain and stiffness, and timed stair climb) following lateral wedge with arch support use (Table 1). More adverse effects were reported with the lateral wedges alone, and more people preferred the lateral wedges with arch support overall (17 out of 22). Finally, though the lateral wedges with arch support were reported to be more comfortable (6.9 ± 2.5 out of 10 vs 6.4 ± 2.6 out of 10), the between-condition difference was not statistically significant. Conclusions: Our study showed that the addition of customized arch support to a standard lateral wedge insole may improve foot and knee symptoms in people with knee OA and concomitant flat feet. Importantly, it was found that these “supported lateral wedges” can be safely delivered to a subgroup of people with knee OA (those with flat feet) for whom lateral wedges may have previously not been advocated. These findings support addressing foot mechanics when designing shoe-worn insoles for treatment of knee OA.
Table 1 Outcome data. Mean (sd) baseline and follow-up outcome data for all variables, as well as model adjusted differences (follow-up minus baseline; beta coefficient (95% confidence intervals)) within each condition as well as between conditions. Model estimate values represent the within- and between-condition least-square mean differences adjusted for period and random subject effects from the regression modeling. Lateral Wedges
Lateral Wedges plus Arch Support
Between-conditions comparison
Variable
Baseline
Follow-up
Difference
Baseline
Follow-up
Difference
Difference
Stair climb (seconds) WOMAC Pain (0-20) WOMAC Function (0-68) FFI Pain(25-100%) FFI Stiffness (25-100%)
5.32 (1.68) 5.9 (3.0) 20.2 (11.0) 41.0 (17.4) 39.2 (15.7)
5.35 (1.54) 5.2 (2.9) 17.6 (8.9) 38.4 (11.9) 37.1 (13.7)
0.04 (0.55, 0.63) 0.7 (1.8, 0.5) 2.5 (6.2, 1.1) 2.7 (8.9, 3.5) 2.1 (7.3, 3.0)
6.57 (4.04) 6.3 (3.4) 21.4 (12.5) 43.6 (16.2) 42.8 (19.5)
5.58 (2.29) 4.0 (2.5) 14.0 (8.8) 33.0 (9.3) 32.3 (9.4)
0.69 (1.3, 0.08)* 1.9 (3.1, 0.6)* 6.1 (9.9, 2.3)* 7.0 (13.5,0.6)* 6.3 (11.7, 0.9)*
0.73 (1.36, 0.11)* 1.2 (2.9, 0.5) 3.6 (7.9, 0.8) 4.3 (12.0, 3.4) 4.1 (11.0, 2.7)
*p < 0.05