AGING IN

EXTRACELLULAR MATRIX
(ECM)
DR. TAN FEI FAN, M.BIOMED
DEPT. HISTOLOGY
 OVERVIEW
CONNECTIVE TISSUE & ECM

CONNECTIVE TISSUE
• 1OF HUMAN BASIC TISSUE ORGANS
• LIES ON EVERY SINGLE OF HUMAN BODY SYSTEMS
• FUNCTION : AS A ‘BINDING” BETWEEN BASIC TISSUE ORGANS
• COMPRISES OF :
• CELLULAR COMPONENT
• NON-CELLULAR COMPONENTS  ECM
 WHAT IS ECM ?
• IT’S A NON CELLULAR COMPONENT OF CONNECTIVE TISSUE &
MAJOR CONSTITUENTS OF THE CONNECTIVE TISSUE
• EACH TISSUE HAS AN ECM WITH A UNIQUE COMPOSITION &
TOPOLOGY THAT IS GENERATED DURING TISSUE DEVELOPMENT
• THROUGH A DYNAMIC & RECIPROCAL, BIOCHEMICAL & BIOPHYSICAL
DIALOGUE BETWEEN THE VARIOUS CELLULAR COMPONENTS
• PROVIDES NOT ONLY ESSENTIAL PHYSICAL SCAFFOLDING FOR THE
CELLULAR CONSTITUENTS BUT ALSO INITIATES CRUCIAL BIOCHEMICAL
& BIOMECHANICAL CUES THAT ARE REQUIRED FOR TISSUE
MORPHOGENESIS, DIFFERENTIATION AND HOMEOSTASIS
 WHAT IS ECM ?
• COMPRISES OF PROTEIN :
• STRUCTURAL  FIBRIL/FIBROUS COMPONENT : COLLAGENS, ELASTIN
• NON STRUCTURAL : FIBRONECTIN, LAMININ, TENASCIN
• OTHERS : INTEGRINS, GROWTH FACTORS (GF) & GROUP OF MATRIX
METALLOPROTEINASE (MMP), WATER
• ANOTHER CLASSIFICATION, ECM COMPRISE OF :
• FIBRIL/FIBROUS COMPONENT : COLLAGENS, ELASTIN
• GROUND SUBSTANCES : GLYCOSAMINOGLYCAN (GAG), PROTEOGLYCAN &
GLYCOPROTEIN
• PLAY A PART IN NUMEROUS CELLULAR PROCESSES INCLUDING CELL
PROLIFERATION, DIFFERENTIATION & MIGRATION
• TO ‘GLUE’ THAT BINDS CELLS TOGETHER IN CONNECTIVE TISSUES
 HEALTHY ECM
• THE CONDITION THAT A FUNCTIONALLY COMPETENT NORMAL TISSUE
CAN EASILY RESIST COMPRESSIVE STRESSES BECAUSE OF THE
BINDING OF THE HYDRATED GAG NETWORK TO THE FIBROUS ECM
MOLECULES

HOW IT WORKS ?
• NON-ACTIVATED TISSUE FIBROBLASTS SECRETE & ORGANIZE TYPE I &
III COLLAGENS, ELASTIN, FIBRONECTIN, TENASCIN & A REPERTOIRE OF
PGS  TO MAINTAIN THE STRUCTURAL & FUNCTIONAL INTEGRITY OF
THE INTERSTITIAL ECM
 HEALTHY ECM (2)
WHY THIS IS POSSIBLE ?
• THE RELAXED NETWORK OF COLLAGEN & ELASTIN FIBERS ALLOW
HEALTHY ECM TO RESIST A WIDE RANGE OF TENSILE STRESSES
• A FUNCTIONALLY COMPETENT NORMAL TISSUE CAN ALSO EASILY
RESIST COMPRESSIVE STRESSES BECAUSE OF THE BINDING OF THE
HYDRATED GAG NETWORK TO THE FIBROUS ECM MOLECULES
• TISSUE ECM IS A HIGHLY DYNAMIC ENTITY THAT CONTINUOUSLY
UNDERGOES REGULATED REMODELING, WHOSE PRECISE
ORCHESTRATION IS CRUCIAL TO THE MAINTENANCE OF NORMAL
FUNCTION
 HEALTHY ECM (3)

TISSUE HOMEOSTASIS
• MEDIATED BY THE COORDINATED SECRETION OF FIBROBLAST MMPS,
THIS IS COUNTERBALANCED BY TIMPS & THE CONTROLLED ACTIVITY
OF OTHER ENZYMES SUCH AS LOX, TRANSGLUTAMINASES THAT
CROSSLINK & STIFFEN THE ECM
• ECM BOUND GFS DIFFERENTIALLY MODULATE CELL GROWTH &
MIGRATION, AND WHEN RELEASED, COMPRISE PART OF TIGHTLY
CONTROLLED FEEDBACK CIRCUIT  ESSENTIAL FOR NORMAL TISSUE
HOMEOSTASIS
 AGING IN ECM
CHARACTERIZED OF AGING ECM
• THINNING OF THE BM, PROBABLY BECAUSE OF ELEVATED MMP MEDIATED
DEGRADATION & REDUCED BM PROTEIN SYNTHESIS
• GROWTH ARRESTED OF RESIDENT FIBROBLASTS IN AGED TISSUE
(TELOMERE SHORTENING) & RESISTANT TO APOPTOTIC CUES  AS
INDICATOR OF SENESCENCE
• A CHRONIC INFLAMMATION
• DISORGANIZED OF FIBROUS COMPONENT OF ECM
• LOSS IN ELASTICITY
• LACK ADAPTIVE RESPONSE TO ENVIRONMENTAL CHANGES
 AGING IN ECM (2)

COMPROMISE THE INTEGRITY OF THE BM

• CHRONIC INFLAMMATION, ELEVATED MMPS, PAI & ROS DESTROY THE
INTEGRITY OF THE ELASTIN NETWORK & MODIFY THE COLLAGEN
FIBER NETWORK  REDUCED LEVEL OF TISSUE-ASSOCIATED GAGS
CHRONIC INFLAMMATION IN AGING
• SENESCENT FIBROBLASTS TYPICALLY EXPRESS ELEVATED LEVEL OF FN,
MMPS, GFS, INTERLEUKIN (IL) & CYTOKINES, AS WELL AS HIGH LEVELS
OF PLASMINOGEN ACTIVATOR INHIBITOR (PAI) & MITOCHONDRIAL
RELATED REACTIVE OXYGEN SPECIES (ROS)
 AGING IN ECM (3)

DISORGANIZED OF FIBROUS COMPONENT OF ECM

• COLLAGEN FIBERS ARE FREQUENTLY INAPPROPRIATELY CROSSLINKED
THROUGH GLYCATION BY PRODUCTS OF LIPID OXIDATION &
THROUGH EXPOSURE TO UV LIGHT
• COLLAGEN TYPE I FIBRES BECOME LESS ORGANIZED, LOOSER &
FRAGMENTED
• CHRONIC INFLAMMATION
 AGING IN ECM (4)

LOSS OF ELASTICITY
• THE ACTION OF MMPS DEGRADING THE MAIN CONSTITUENTS OF
ECM SUCH AS TYPE I & III COLLAGEN, PROTEOGLYCANS & GAGS
• TOGETHER WITH INCREASED CROSSLINKING OF COLLAGEN 
REDUCE BIOMECHANICAL PROPERTIES OF THE TISSUE
LACK ADAPTIVE TO ENVIRONMENTAL CHANGES
• ALTERATION OF FN EXPRESSION HAS A DETRIMENTAL CHANGE,
ESPECIALLY WITH AGE
 AGING IN ECM (5)

FINAL RESULT
• TISSUE STIFFENING
• MECHANICALLY WEAKER
• LESS ELASTIC
• RIGIDITY
• COMPROMISE ECM ORGANIZATION
• MODIFY EPITHELIAL ORGANIZATION & FUNCTION
• POTENTIALLY PROMOTING AGE-RELATED DISEASE, SUCH AS CANCER
 FIBROSIS
• USUALLY COME AFTER INJURY
• TISSUE STIFFNESS
HOW THIS HAPPENED?
• UPON BINDING TO ECM DEGRADATION PRODUCTS & CYTOKINES, MONOCYTES
RAPIDLY DIFFERENTIATE INTO MACROPHAGES
• THIS ACTIVATED MACROPHAGE SECRETE & RELEASE MULTIPLE GFS, MMPS &
CYTOKINES THAT PROMOTE ANGIOGENESIS & STIMULATE FIBROBLAST
MIGRATION & PROLIFERATION
• RECRUITED FIBROBLAST BEGIN TO SYNTHESIZE & DEPOSIT LARGE QUANTITIES OF
ECM PROTEINS INCLUDING COLLAGEN TYPE I & III, FN, HYALURONIC ACID (HA)
• ELEVATED MECHANICAL STRESS ASSOCIATED WITH THIS PROFOUND ECM
DEPOSITION CAN INDUCE TRANSDIFFERENTIATION OF FIBROBLASTS & OTHER
TISSUE RESIDENT CELLS (EX : EPITHELIAL TO MESENCHYMAL TRANSITION ;
CIRCULATING BONE MARROW DERIVED MESENCHYMAL STEM CELL TO
MYOFIBROBLAST)
 SCARRED TISSUE

• BESIDE OF FIBROSIS IT SELF, MYOFIBROBLAST PLAYS ROLE

• MYOFIBROBLAST PROMOTE LARGE, RIGID COLLAGEN BUNDLES THAT
CROSSLINKED BY LOX ENZYMES  STREGHTEN & STIFFEN THE TISSUE
• IMBALANCE OF TIMPS & MMP SYNTHESIS CAN PROMOTE CHRONIC
VASCULAR REMODELING & ENHANCE ECM CROSSLINKING 
ABBERANT FIBROSIS & INSTABILITY MECHANICAL PROPERTIES
(ELASTICITY)
 TUMOR & CANCER
• STIFFENING OF TUMORS IS INDUCED BY ECM DEPOSITION & REMODELING BY RESIDENT
FIBROBLASTS, ALSO INCREASED CONTRACTILITY THE TRANSFORMED EPITHELIUM
• CHEMOKINES & GFS INDUCED INFLAMMATION & MODIFIED REPERTOIRE OF
INFILTRATING T LYMPHOCYTES
• THIS INFLAMMATION CHANGE FIBROBLAST INTO MYOFIBROBLAST  EXACERBATING &
PROMOTING TISSUE DESMOPLASIA
• MMPS ARE SECRETED & ACTIVATED BY MYOFIBROBLAST & TUMOR CELLS
• REMODELLING OF BM SURROUNDING THE TUMOR  RELEASE & ACTIVATED ECM
EMBEDDED – GFS
• RELEASED OF VEGF ENHANCES VASCULAR PERMEABILITY & PROMOTE NEW BLOOD
VESSEL GROWTH
• INDUCE ANGIOGENESIS, INVASION & EVENTUALLY FOSTER MESTASTASIS.
 DISEASES RELATED TO AGING ECM

• DIABETIC NEPHROPATHY
• ATHEROSCLEROSIS
• HYPERTENSION
• SKIN AGING & WOUND HEALING
 DIABETIC NEPHROPATHY (DN)
• A SERIOUS COMPLICATION IN DIABETES.
• MAJOR TYPICAL MORPHOLOGICAL CHANGES ARE THE RESULT OF
CHANGES IN THE EXTRACELLULAR MATRIX (ECM).
• THICKENED OF BM
• THE GLOMERULAR MESANGIAL MATRIX AND THE TUBULOINTERSTITIAL SPACE
ARE EXPANDED, DUE TO INCREASED AMOUNTS OF ECM.
• ONE IMPORTANT ECM COMPONENT, THE PROTEOGLYCANS (PGS), SHOWS
A MORE COMPLEX PATTERN OF CHANGES IN DN. PGS IN BASEMENT
MEMBRANES ARE DECREASED BUT INCREASED IN THE MESANGIUM AND THE
TUBULOINTERSTITIAL SPACE.
• THE AMOUNTS AND STRUCTURES OF HEPARAN SULFATE CHAINS ARE
CHANGED, AND SUCH CHANGES AFFECT LEVELS OF GROWTH FACTORS
REGULATING CELL PROLIFERATION AND ECM SYNTHESIS, WITH CELL
ATTACHMENT AFFECTING ENDOTHELIAL CELLS AND PODOCYTES.
 DIABETIC NEPHROPATHY (DN) (2)

• ENZYMES MODULATING HEPARAN SULFATE STRUCTURES, SUCH AS

HEPARANASE AND SULFATASES, ARE IMPLICATED IN DN.
• OTHER ENZYME CLASSES ALSO MODULATE ECM PROTEINS AND PGS,
SUCH AS MMPS & SERINE PROTEASES, SUCH AS PLASMINOGEN
ACTIVATOR, AS WELL AS THEIR CORRESPONDING INHIBITORS. THE
LEVELS OF THESE ENZYMES AND INHIBITORS ARE CHANGED IN PLASMA
AND IN THE KIDNEYS IN DN.
• SEVERAL GROWTH FACTORS, SIGNALING PATHWAYS, AND
HYPERGLYCEMIA PER SE AFFECT ECM SYNTHESIS AND TURNOVER IN DN.
 ATHEROSCLEROSIS
• RECENT STUDIES SUGGEST THAT STIFFENING IS AN IMPORTANT CLINICAL TARGET
NOT ONLY BECAUSE OF POTENTIAL DELETERIOUS EFFECTS ON THE HEART BUT
ALSO BECAUSE IT PROMOTES CELLULAR LEVEL DYSFUNCTION IN THE VESSEL WALL,
CONTRIBUTING TO A PATHOLOGICAL ATHEROSCLEROTIC STATE.
• ARTERIAL STIFFENING OCCURS WITH AGE AND IS CLOSELY ASSOCIATED WITH
THE PROGRESSION OF CARDIOVASCULAR DISEASE.  HEART FAILURE
• RECENT EVIDENCE SUGGESTS THAT THE IMPACT OF INCREASED VESSEL
STIFFENING EXTENDS BEYOND THE TISSUE AND CAN ALSO HAVE DELETERIOUS
EFFECTS ON CELLULAR FUNCTION.
• ALTERED EXTRACELLULAR MATRIX (ECM) ARCHITECTURE HAS BEEN RECOGNIZED AS
A KEY COMPONENT OF THE PRE-ATHEROGENIC STATE.
• THE UNDERLYING CAUSES OF AGE-RELATED VESSEL STIFFENING ARE AGE-RELATED
CROSSLINKING OF THE ECM PROTEINS AS WELL AS THROUGH INCREASED MATRIX
DEPOSITION.
 ATHEROSCLEROSIS (2)
• THE COLLAGEN CONCENTRATION IN ALL THREE LAYERS OF THE ARTERIAL WALL
INCREASES WITH AGE, SHIFTING THE ELASTIN:COLLAGEN BALANCE THAT GOVERNS
HEALTHY ARTERIAL MECHANICS.
• MEDIAL FIBROSIS OCCURS AS A CONSEQUENCE OF COLLAGEN FIBERS REPLACING
VASCULAR SMOOTH MUSCLE CELL (VSMCS)
• ELASTIC FIBERS HAVE AN EXTREMELY LOW TURNOVER RATE IN VIVO, AND THIS
LONGEVITY ALLOWS FOR THE ACCUMULATION OF AGE-RELATED CHANGES CAUSED BY
FRAGMENTATION, CALCIFICATION, AND MMP-DEGRADATION
• AS ELASTIN FIBERS DECAY, THEY LOSE FUNCTIONALITY AND SHIFT LOAD BEARING
ONTO STIFFER COLLAGEN FIBRILS, WHICH DIRECTLY CONTRIBUTES TO SIGNIFICANT
INCREASES IN ARTERIAL STIFFNESS.
• CALCIUM IN THE ARTERIAL WALL ALSO INCREASES WITH AGE FACILITATING THE DIRECT
BINDING OF CALCIUM IONS TO ELASTIN FIBERS CAUSING CALCIFICATION
 ATHEROSCLEROSIS (3)

• INCREASED ACTIVITY OF THE ELASTASES MT1-MMP AND MMP-2 HAS

BEEN OBSERVED, AND MMP-2 HAS BEEN FOUND NEAR FRAGMENTED
ELASTIN FIBERS WITHIN THE AORTA
• THE DYSREGULATION OF MMPS IS ALREADY KNOWN TO PLAY A ROLE IN
THE CARDIOVASCULAR PATHOLOGIES HYPERTENSION AND ANEURYSM
• MMP-2 IS FOUND NEAR FRAGMENTED ELASTIN FIBERS WITHIN THE
AORTA. NOTABLY, EVEN THOUGH THE ABSOLUTE ELASTIN CONTENT IN
THE AORTA REMAINS RELATIVELY STABLE WITH AGE, THE ELASTIN
CONCENTRATION DECREASES AND IS ACCOMPANIED BY A SUBSTANTIAL
INCREASE IN COLLAGEN CONCENTRATION
 ATHEROSCLEROSIS (4)

• THE CONCENTRATIONS OF DESMOSINE AND ISODESMOSINE AND

THEIR CROSSLINKS DECREASE WITH AGE. THE COMPOUNDS
DESMOSINE AND ISODESMOSINE ARE FORMED FROM 4 LYSINE
AMINO ACIDS AND ARE CRITICAL FOR CROSSLINKING ELASTIN FIBERS
TO GIVE THEM THEIR ELASTIC PROPERTIES
 ATHEROSCLEROSIS (5)

• AGED ENDOTHELIAL CELL (ECS) HAVE MORPHOLOGICAL CHANGES RESEMBLING A

VSMC PHENOTYPE INDICATING DEPOSIT COLLAGEN THAT CONTRIBUTES TO
INTIMAL THICKENING.
• WITHIN THE ADVENTITIA, COLLAGEN I AND III DEPOSITION BY FIBROBLASTS
INCREASES WITH AGE AND IS ACCOMPANIED BY VESSEL STIFFENING.
• IN CONCERT WITH INCREASED COLLAGEN CONCENTRATIONS, COLLAGEN
CROSSLINKING BY NON-ENZYMATIC GLYCATION INCREASES ARTERIAL STIFFNESS
WITH AGE. GLYCATION IS A REACTION BETWEEN REDUCING SUGARS AND
PROTEINS, AND DIRECTLY STIFFENS TISSUES IN ADDITION TO PRODUCING
DELETERIOUS END PRODUCTS ADVANCED GLYCATION END PRODUCTS (AGES)
 ATHEROSCLEROSIS (6)

• AGES ARE HARMFUL TO VASCULAR HEALTH, BY REDUCING NITRIC

OXIDE (VASODILATOR). NITRIC OXIDE (NO) CAN MAINTAIN VASCULAR
TONE & HAS ANTI-INFLAMMATORY EFFECTORS ON THE ENDOTHELIUM
• AGES INTERACT WITH THE RECEPTOR FOR ADVANCED GLYCATION
END PRODUCTS (RAGE) TO HAVE DOWNSTREAM EFFECTS THAT
INCLUDE THE PRODUCTION OF REACTIVE OXYGEN SPECIES, NF-ΚB
INFLAMMATORY SIGNALING, AND ENDOTHELIAL HYPERPERMEABILITY
THE END