Tissue engineering with meniscus cells derived from surgical debris

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Tissue engineering with meniscus cells derived from surgical debris B.M. Baker, B.S., A.S. Nathan, G. Russell Huffman, M.D., M.P.H., R.L. Mauck, Ph.D. Osteoarthritis and Cartilage Volume 17, Issue 3, Pages 336-345 (March 2009) DOI: 10.1016/j.joca.2008.08.001 Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 1 Morphological appearance of passage 2 human MDCs in monolayer and on fiber-aligned nanofibrous scaffolds. (A) Passage 2 MDCs in monolayer on tissue culture polystyrene demonstrate a fibroblast-like morphology. (B) Passage 2 MDC-seeded constructs cultured for 1 day reveal MDCs elongating in and aligning with the predominant fiber direction of the scaffold. Red: F-actin, white: fibers, blue: nuclei. Scale bar: 50μm. Osteoarthritis and Cartilage 2009 17, 336-345DOI: (10.1016/j.joca.2008.08.001) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 2 MDC-seeded scaffolds increase in mechanical properties with time in culture in a fibrocartilaginous medium. (A) Force–elongation plots of five scaffolds either seeded (MDC) or maintained as USCs on day 70 for Donor 8. (B) Maximum load of seeded scaffolds normalized to that of paired USC scaffolds on day 70 for all 10 donors. Donor # is indicated on the x-axis. Data represent the mean and standard deviation of five samples per donor. (C) Normalized stiffness (indicating percentage change) of MDC-seeded scaffolds from each donor compared to their paired USC scaffolds at each time point. Donor # (and age) is indicated on the x-axis. Osteoarthritis and Cartilage 2009 17, 336-345DOI: (10.1016/j.joca.2008.08.001) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 3 Donor-to-donor variation in time-dependent changes in biochemical composition of MDC-seeded engineered meniscus constructs. (A) Total collagen and (B) total s-GAG accumulation in engineered constructs with time in culture for each donor. Donor # (and age) is indicated on the x-axis. Data represent the mean and standard deviation of five samples per donor per time point. (C) Percent DW (% DW) collagen and (D) % DW s-GAG for samples from each of the 10 donors on day 70. Gray background in (C) and (D) indicates range of collagen and s-GAG found in five native tissue samples. Note the interrupted scale in (C) the % collagen by DW plot. USC constructs processed similarly showed no appreciable background content of s-GAG or collagen. Osteoarthritis and Cartilage 2009 17, 336-345DOI: (10.1016/j.joca.2008.08.001) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 4 Structure–function-composition correlations for MDC-seeded constructs with time in culture. (A) Total collagen content in constructs correlates well with change in stiffness for all donors at all time points (days 14, 42, and 70). (B) Total GAG content correlates poorly with change in stiffness for all donors at all time points (days 14, 42, and 70). (C) Donor age showed a weak correlation with change in stiffness of constructs on day 70. Osteoarthritis and Cartilage 2009 17, 336-345DOI: (10.1016/j.joca.2008.08.001) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions

Fig. 5 Histological analysis of constructs from best-case (Donor 8, A,C,E) and worst-case (Donor 6, B,D,F) samples on day 70. DAPI-staining of cell nuclei demonstrate infiltration into the outer two-thirds of constructs under best-case conditions (A), and limited infiltration at the periphery under worst-case conditions (B). Similar findings are observed for collagen (C,D) and proteoglycan (E,F) deposition as indicated by PSR and AB staining, respectively. Scale bar: 1mm. Osteoarthritis and Cartilage 2009 17, 336-345DOI: (10.1016/j.joca.2008.08.001) Copyright © 2008 Osteoarthritis Research Society International Terms and Conditions