Advanced Regenerative Medicine

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Ageing

With continuously increasing life expectancy, the impact of chronic diseases and dependencies, for which no specific cures are available, also increases. This leads to an enormous burden to individuals and societies alike, which threatens to overwhelm conventional curative and care models in medicine. Unfortunately, healthy life expectancy, the number of years spent free of disabilities, has grown slower than life expectancy in a number of countries. Within the European Union, for instance, life expectancy at age 50 has grown by 1.2 years between 2005 and 2010, but healthy life expectancy has only grown slightly (0.5 years in men, 0.4 years in women) in the same time frame (Fouweather et al., 2015).

Aging is a process of progressive decline in the physiological capacity of an organism, manifested by accumulated alteration and destabilization at the whole system level. Generally, oxidative stress is a cause of aging in lipid peroxidation, protein carbonyl content increase, and DNA damage. Aging is also associated with a differential gene expression pattern indicative of a marked or lower stress response of metabolic and biosynthetic genes. Additionally, cellular senescence, stem cell exhaustion and chronic low-grade inflammation known as “inflammaging” have been proposed as limitations to age span and health span.

Mesenchymal stem cells and their secretome

Mesenchymal stem cells, or MSCs, are multipotent stromal cells that can differentiate into a variety of cell types, including: osteoblasts, chondrocytes, neurons, muscle cells and adipocytes. This phenomenon has been documented in specific cells and tissues in vivo and in vitro. MSCs are distributed all over the body and are responsible for regeneration. Commonly used tissues for the isolation of MSC are bone marrow, umbilical cord, cord lining and, increasingly, adipose tissue, which has a superior amount of MSCs.

All cells communicate and exchange information by different ways, including the secretion of soluble factors, the cell-to-cell adhesion contact, and the intercellular exchange of organelles. The secretome of mesenchymal stem cells includes paracrine substances, exosomes and microvesicles. Over 150 paracrine substances, also called cytokines and chemokines, can be released by mesenchymal stem cells. Two distinct populations of vesicles with peculiar membrane structure, mechanism of production, pathophysiological relevance, and different size

have been described: exosomes and microvesicles. Microvesicles and exosomes contain biomolecules, including messenger RNA and micro RNA. The secretome of adipose derived stem cells has a predominantly anti-inflammatory and anti-apoptic effect and target tissues.

Saliva biomarkers for anti-ageing

BACE 1 (Beta site amyloid precursor protein cleaving enzyme number one)

enzyme responsible for the production of amyloid beta deposits in Alzheimer’s disease, Age related macular degeneration, Diabetes type 2, ALS, Parkinson Disease. High levels are bad for your health.

UCH-L1 (Ubiquitin carboxyl-terminal hydrolase L1):

main enzyme in the clearance of toxic waste products such as amyloid beta, improves memory deficits. Low levels means your internal waste disposal system is not working. However, very high levels may be associated with brain injury!

Phosphorylated α-synuclein

misfolding and abnormal accumulation of α-synuclein play a significant role in the pathogenesis of Parkinson Disease (PD) and leads to death of neurons. High levels of α-synuclein are signs of PD or risk of developing PD.

GPNMB (glycoprotein nonmetastatic melanoma protein B)

Because of its various involvement in different physiological conditions, GPNMB has been implicated in many diseases: Alzheimer's Disease, Parkinson's Disease, Cancer formation, Accumulation of senescent cells and Ageing.

08-OHdG DNA damage (8-hydroxy-2’deoxy-Guanosine):

8-OHdG is a marker of oxidative stress and is associated with DNA damage (shortening of telomeres). High levels mean high risk of DNA damage.

Regenerative proteins secreted by mesenchymal stem cells

The secretome of MSCs contains hundreds of active regenerative proteins with antiageing properties, such as neprilysin (NEP), galectin-1 and Ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1).

NEP is member of the M13 zinc metallopeptidase family. NEP is primarily expressed in the kidneys; however, it occurs at much lower levels in many other tissues, including brain. Neprilysin occurs naturally in the secretome of the ad-MSC, which is produced in vitro and in vivo as a means of communication from cell to cells and as a reaction to external influences. NEP binds to amyloid beta oligomers and preventing them from being deposited.

Galectin 1 is produced naturally by mesenchymal stem cells. Galectin 1 is a galactose binding lectin. It blocks alfa synuclein at the cell membrane and prevents it from being internalized. This will prevent the main pathological pathway in Parkinson’s disease.

Ubiquitin carboxyl-terminal hydrolase L1 is a member of the deubiquitinating enzymes DUB. UCH-L1 removes ubiquitin from ubiquinated substances thus enabling degradation of the target enzyme, as well as recycling and stabilizing free ubiquitin. UCH-L1 accelerates BACE1 degradation, regulates Amyloid Precursor Protein and reduces Amyloid beta production in Alzheimer Disease and Spinal Cord injury, inhibits neuritic plaque formation and improves memory deficits (in rodent model). High concentrations are related to increased death of brain cells.

Clearance of aggregated and oxidized proteins

Generally, MSCs and the secretome have a regenerative mode of action, influencing all body systems via activation of systems responsible for the clearance of deposits of aggregated and oxidized proteins which accumulate over the lifespan of cells. This accumulation results both from increased occurrence of damage through oxidative stress and the decrease of clearance through degradation processes. The main clearance (70-80%) is via the ubiquitin proteasome system UPS.

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