Massachusetts Institute of Technology RT Harvard medical school Brigham and Womens/Massachusetts General Hosp VA Boston Healthcare System 2.79J/3.96J/BE.441/HST522J CELL-MATRIX INTERACTIONS M. Spector, Ph. D
Massachusetts In Massachusetts Institute of Technology stitute of Technology Harvard Medical School Harvard Medical School Brigham and Wo Brigham and Women’s/Massachusetts General Hosp. men’s/Massachusetts General Hosp. VA Boston Healthcare System VA Boston Healthcare System 2.79J/3.96J/BE.441/HST522J 2.79J/3.96J/BE.441/HST522J CELL-MATRIX INTERACTIONS MATRIX INTERACTIONS M. Spector, Ph.D. M. Spector, Ph.D
Articular Cartilage Extracellular Matrix Cell 4 mm e 10 um
Cell Extracellular Matrix 4 mm 10 µm Articular Cartilage
Chondrocytes(p2 canine) in a Type I Collagen-GAG Scaffold Image removed due to copyright considerations “ Control volume B. Kinner
Chondrocytes (P2 Canine) in a Type I Collagen-GAG Scaffold Image removed due to copyright consi derati ons. “Control Volume” B. Kinner
UNIT CELL PROCESSES Concept of a“ Control volume” around a cell Soluble Regulator Cell Matrix Product+ Soluble (Regulator) Regulator b “ Control volume
UNIT CELL PROCESSES UNIT CELL PROCESSES Concept of a “Control Volume” around a Cell Concept of a “Control Volume” around a Cell Cell + Matrix Cell + Matrix Product + Soluble Product + Soluble (Regulator) (Regulator) Regulator B Regulator B “Control Volume” “Control Volume” Soluble Soluble Regulator Regulator A
COLLAGEN-GAG MATRICES: MODEL BIOMATERIALS (ANALOGS OF EXTRACELLULAR MATRIX) Investigation of cell interactions (UCPs)in vitro Type I(bovine and porcine) Type II(porcine) Image removed due to Chondroitin 6-sulfate copyright considerations Imm Image removed due to copyright considerations Freeze-dried Dehydrothermally cross-linked Additional cross-linking 500um IV Yannas, et aL. PNas. 1989
COLLAGEN-GAG MATRICES: MODEL BIOMATERIALS (ANALOGS OF EXTRACELLULAR MATRIX) Investigation of cell interactions (UCPs) in vitro • Type I (bovine and porcine) • Type II (porcine) • Chondroitin 6-sulfate Image removed due to copyright consi derati ons. 1mm Image removed due to copyright consi derati ons. • Freeze-dried • Dehydrothermally cross-linked • Additional cross-linking 500 µ m IV Yannas, et al. PNAS, 1989
CELL-MATRIX INTERACTIONS WITH COLLAGEN-GAG MATRICES IN VITRO Cell interactions with the scaffold Mitosis, migration, synthesis, and contraction, and their interrelationships Can provide insights into scaffold composition and structure for improved performance in regenerative medicine Can provide insights into cell behavior
CELL –MATRIX INTERACTIONS WITH MATRIX INTERACTIONS WITH COLLAGEN COLLAGEN-GAG MATRICES GAG MATRICES IN VITRO IN VITRO • Cell interactions with the scaffold Cell interactions with the scaffold – –Mitosis, migration, synthesis, and Mitosis, migration, synthesis, and contraction, and their interrelationships contraction, and their interrelationships • Can provide insights into scaffold Can provide insights into scaffold composition and structure for improved composition and structure for improved performance in regenerative medicine performance in regenerative medicine • Can provide insights into cell behavior Can provide insights into cell behavior in vivo in vivo
Chondrocytes(Passage 2 Canine) in a Type I Collagen-GAG Matrix Live cell imaging for a period of 5 hours Image removed due to copyright considerations J. Cheng
Chondrocytes (Passage 2 Canine) in a Type I Collagen-GAG Matrix Live cell imaging for a period of 5 hours. Image removed due to copyright consi derati ons. J. Cheng
CELL-MATRIX INTERACTIONS Mitosis Migration ynthesis Contraction
CELL –MATRIX INTERACTIONS MATRIX INTERACTIONS • Mitosis Mitosis • Migration Migration • Synthesis Synthesis • Contraction Contraction
Chondrocyte(p2 Canine) in a Type I Collagen-GAG Matrix: Mitosis Image removed due to copyright considerations J. Cheng
Chondrocyte (P2 Canine) in a Type I Collagen-GAG Matrix: Mitosis Image removed due to copyright consi derati ons. J. Cheng
Effects of Cross- Linking on Chondrocyte proliferation in Type I Collagen-GAG Matrices 60 Mean±SEM 50 Inc. X-lInking 40 -DHT UV 30 -GTA EDAC 20 10 0 10 20 30 Day CR Lee, et al. Biomat. 2001: 22 3145
DNA Content 0 10 20 30 40 50 60 0 10 20 30 Day DNA ( u g ) DHT UV GTA EDAC CR Lee, et al., Biomat. 2001;22:3145 Mean ± SEM Effects of Cross Effects of Cross-Linking on Chondrocyte Proliferation Linking on Chondrocyte Proliferation in Type I Collagen in Type I Collagen-GAG Matrices GAG Matrices Inc. x-linking linking