Keratocyte matrix interactions and thrombospondin 2

[摘要] Purpose: To determine whether human keratocytes synthesizethrombospondins 2 and 3 (TSP-2, TSP-3) in a collagen matrix and theeffect of addition of antibodies to TSP-2 and TSP-3 onkeratocyte-populated collagen matrix contraction.Methods: Keratocyte-populated collagen matrices were evaluated forTSP-2 and TSP-3 mRNA. Sections of matrices were stained byimmunohistochemistry for TSP-2 and TSP-3. Keratocyte populated collagenmatrices were treated with antibodies specific for TSP-2 and TSP-3 andthese preparations were evaluated for contraction and keratocytemorphology.Results: Keratocyte derived fibroblasts in collagen matricescontained TSP-2 and TSP-3 mRNA, and the cells wereimmunoreactive for both proteins. Compared to controls, an antibodyspecific to the N-terminal domain of TSP-2 significantly inhibitedmatrix contraction for up to 10 days at a concentration of 20 μg/mlantibody. At 2 μg/ml TSP-2 antibody concentration significantinhibition occurred up to 3 days. Removal of the antibody from the mediareversed the inhibitory effect. Cultured keratocytes in TSP-2 treatedcollagen matrices appeared more rounded than keratocytes in controlmatrices. An antibody specific to TSP-3 had no effect on matrixcontraction or keratocyte morphology.Conclusions: Keratocyte derived fibroblasts synthesize TSP-2 andTSP-3 when seeded in collagen matrices. Antibody specific to TSP-2reversibly inhibits matrix contraction. TSP-2 may play a key role inkeratocyte/collagen matrix interactions, as may occur during cornealstromal repair.

[学科分类] 生物化学/生物物理