Nicholas P. Ziats, PhD, James M. Anderson, MD, PhD

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Presentation transcript:

Human vascular endothelial cell attachment and growth inhibition by type V collagen Nicholas P. Ziats, PhD, James M. Anderson, MD, PhD Journal of Vascular Surgery Volume 17, Issue 4, Pages 710-718 (April 1993) DOI: 10.1016/0741-5214(93)90115-3 Copyright © 1993 Society for Vascular Surgery and International society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 1 Effects of human collagen type V or fibronectin coated on ePTFE and attachment of HSVECs. Human type V collagen (+V) or fibronectin (+FN) at 10 μg/ml (0.35 ml/polymer) was coated for 1 hour at 37° C and rinsed, and cells were added. FN > V refers to coating with fibronectin first, then type V collagen. V > FN refers to coating with type V collagen first, then fibronectin. FN + V refers to type V and fibronectin mixed in solution and then added to the surface. HSVECs were initially plated (solid circle) at 95,000 cells/polymer, and attachment was evaluated after 18 to 24 hours. NONE refers to no protein coating. Bar graphs represent mean value of triplicate wells ± SD. Fibronectin coating (+FN)—enhanced attachment (p < 0.05) compared with no coating (NONE); type V collagen attachment was significantly reduced (p < 0.025) compared with fibronectin. Journal of Vascular Surgery 1993 17, 710-718DOI: (10.1016/0741-5214(93)90115-3) Copyright © 1993 Society for Vascular Surgery and International society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 2 Effect of human type V collagen (+ TYPE V) or fibronectin (+ FN) on HSVEC (A) or HUVEC (B) attachment and growth on ePTFE. A, HSVECs were initially plated (solid circle) at 95,000 cells/polymer, and attachment (day 0) was evaluated after 18 to 24 hours. Cell growth (days 4 and 7) was evaluated after 24 hours of adherence; cells were rinsed and fed fresh medium every 2 to 3 days. Bar graphs represent mean value of triplicate wells ± SD. NONE refers to no coating or no addition. Tissue culture-treated polystyrene coated with fibronectin (TCPS + FN) is positive control. For HSVECs, type V collagen growth is significantly inhibited at days 4 and 7 when compared with fibronectin (p < 0.05 ) or tissue culture polystyrene (day 4, p < 0.05; day 7, p < 0.025). B, HUVECs were plated (solid circle) at density of 75,000 cells/polymer, and attachment and growth were evaluated as in A. Type V collagen significantly inhibited cell growth compared with fibronectin coating or TCPS + FN at day 7 (p < 0.025). Journal of Vascular Surgery 1993 17, 710-718DOI: (10.1016/0741-5214(93)90115-3) Copyright © 1993 Society for Vascular Surgery and International society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 3 Effect of human type V (+ TYPE V) or IV (+ TYPE IV) collagen and fibronectin (+ FN) on HSVEC attachment and growth on tissue culture-treated polystyrene. HSVECs were initially plated (solid circle) at 70,000 cells/polymer, and attachment (day 0) was evaluated after 18 to 24 hours. Cell growth (days 4 and 7) was evaluated after 24 hours of adherence; cells were rinsed and fed fresh medium every 2 to 3 days. Bar graphs represent mean value of triplicate wells ± SD. NONE refers to no coating or no addition. Type V collagen growth is significantly inhibited at day 4 (p < 0.01) and day 7 (p < 0.05) when compared with fibronectin or tissue culture polystyrene (day 7, p < 0.05). V collagen growth is significantly inhibited at days 4 (p < 0.05) and day 7 (p < 0.025) when compared with type IV collagen. Journal of Vascular Surgery 1993 17, 710-718DOI: (10.1016/0741-5214(93)90115-3) Copyright © 1993 Society for Vascular Surgery and International society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 4 Morphology of HSVECs on fibronectin (A) or type V collagen (B) coated at 10 μg/ml on tissue culture polystyrene plates. Photographs were taken at day 7 (see Fig. 3). (Original magnification × 100.) Journal of Vascular Surgery 1993 17, 710-718DOI: (10.1016/0741-5214(93)90115-3) Copyright © 1993 Society for Vascular Surgery and International society for Cardiovascular Surgery, North American Chapter Terms and Conditions

Fig. 5 Effect of human collagen type V or human fibronectin on human fibroblast attachment and growth on tissue culture-treated polystyrene (A) or ePTFE (B). Attachment (day 0) and growth of human fibrobasts (days 4 and 7) initially plated (solid circle) at 25,000 cells/well on human fibronectin (+ FN) or type V collagen (coated at 10 μg/ml for 1 hour at 37° C). Cells were fed every 2 to 3 days with DMEM with 5% FBS. Journal of Vascular Surgery 1993 17, 710-718DOI: (10.1016/0741-5214(93)90115-3) Copyright © 1993 Society for Vascular Surgery and International society for Cardiovascular Surgery, North American Chapter Terms and Conditions