Altered proliferative responses of dermal fibroblasts to TGF-β1 may contribute to chronic venous stasis ulcer1 Brajesh K Lal, MD, Satoshi Saito, MD,

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Altered proliferative responses of dermal fibroblasts to TGF-β1 may contribute to chronic venous stasis ulcer1 Brajesh K Lal, MD, Satoshi Saito, MD, Peter J Pappas, MD, Frank T Padberg, MD, Joaquim J Cerveira, MD, Robert W Hobson, MD, Walter N Durán, PhD Journal of Vascular Surgery Volume 37, Issue 6, Pages 1285-1293 (June 2003) DOI: 10.1016/S0741-5214(02)75295-6

Fig 1 Dermal fibroblasts were cultured from lower extremity skin biopsy specimens (see Methods). In 2 to 3 weeks, patient fibroblasts could be seen migrating from the explant and populating the culture dish. Journal of Vascular Surgery 2003 37, 1285-1293DOI: (10.1016/S0741-5214(02)75295-6)

Fig 2 Logarithmic growth phase kinetics. Proliferation response of fibroblasts to 5 ng/mL of TGF-β1 over 10 days. A, Response of HS68 fibroblasts. B, Response of fibroblasts from a patient with CEAP class 6 CVI. On linear regression analysis the logarithmic period of growth extended from days 2 to 7 in both cases. Journal of Vascular Surgery 2003 37, 1285-1293DOI: (10.1016/S0741-5214(02)75295-6)

Fig 3 Dose response. Proliferation response of HS68 fibroblasts to increasing concentrations of TGF-β1. Fibroblasts responded with increased proliferation at TGF-β1 concentrations of 2.5 ng/mL (P < .05), 5 ng/mL (P < .001), and 10 ng/mL (P < .01). Journal of Vascular Surgery 2003 37, 1285-1293DOI: (10.1016/S0741-5214(02)75295-6)

Fig 4 Baseline unstimulated proliferation of fibroblasts derived from HS68 cell line, NC skin, and patients with CVI. LT and LC fibroblasts from patients with CEAP 2-3 disease demonstrated proliferation similar to that of HS68 and NC adult skin fibroblasts. CEAP 4 LC fibroblasts proliferated less compared with LT cells (∗P < .001) and HS68 cells (♣P < .05). LC CEAP 5 fibroblasts proliferated less than LT cells did (♦P < .001). LC fibroblasts from patients with CEAP 4 and 5 CVI proliferated to the same degree as HS68 did. However, both LT and LC fibroblasts from CEAP 6 CVI proliferated less than did HS68 and NC adult skin cells (#P < .001 for both). Journal of Vascular Surgery 2003 37, 1285-1293DOI: (10.1016/S0741-5214(02)75295-6)

Fig 5 Proliferation response to TGF-β1 (5 ng/mL) of fibroblasts derived from HS68 cell line, NC skin, and patients with CVI. HS68 cells (∗P < .001), LT fibroblasts (#P < .001), and LC fibroblasts (+P = .007) from patients with CEAP 2-3 CVI and LC fibroblasts (♦P = .004) from patients with CEAP 4 CVI all demonstrated increased proliferation when exposed to TGF-β1. However, LC and LT fibroblasts from patients with CEAP 5 and 6 CVI did not exhibit increased proliferation in response to TGF-β1. Journal of Vascular Surgery 2003 37, 1285-1293DOI: (10.1016/S0741-5214(02)75295-6)

Fig 6 Proliferation response of fibroblasts grown on plates coated with polystyrene, collagen, and fibronectin. A, Without TGF-β1. HS68 fibroblasts demonstrated increased proliferation when grown on collagen (∗P < .01) and fibronectin (♣P < .001) compared witt polystyrene. Proliferation of NC, LC, and LT fibroblasts in any CVI class was not increased by these ECM proteins when compared with growth on polystyrene. B, With TGF-β1 (5 ng/mL). TGF-β1 increased proliferation of HS68, NC, and CEAP 2-3 CVI fibroblasts when cells were cultured on collagen and fibronectin to the same extent as when these cells were grown on polystyrene. TGF-β1 did not increase proliferation of CEAP 4-6 LC fibroblasts grown on collagen or fibronectin, but the response was similar to that of cells grown on polystyrene. LC CEAP 4 fibroblast proliferation was inhibited in the presence of fibronectin (∗P < .05) compared with cells cultured on polystyrene. Journal of Vascular Surgery 2003 37, 1285-1293DOI: (10.1016/S0741-5214(02)75295-6)