VITAMIN C AND SKIN INTERGRITY: MECHANISM OF ACTION AND CLINICAL

IMPLICATIONS

A SPECIAL SEMINAR TOPIC (BCM 4101)

WRITTEN BY:
IKPI, EMMANUEL EYONG
17/BCM/142

SUBMITTED TO
THE DEPARTMENT OF MEDICAL BIOCHEMISTRY
FACULTY OF BASIC MEDICAL SCIENCES
UNIVERSITY OF CROSS RIVER STATE (UNICROSS)
OKUKU CAMPUS

IN PARTIAL FULFILMENT OF THE AWARD OF BACHELOR OF SCIENCE

(B.Sc.) DEGREE IN MEDICAL BIOCHEMISTRY (BCM 4101)

OCTOBER, 2021.
 DEDICATION

This seminar is dedicated to God almighty for his guidance and protection throughout the course
of this study.
 ACKNOWLEDGEMENT

My profound gratitude goes to the Almighty God for divine power in guiding and protecting me
throughout the period of this seminar work.

My profound gratitude goes to my supervisor; Dr. Ugwu, Melvin Nnemeka for taking time to
correct this work and making it fruitful and correct. I also appreciate my humble Head of the
Department, Dr. Ugwu, Melvin Nnaemeka, for his tireless commitment and contribution
towards the excellence of the Department. I also wish to appreciate all the lecturers in the
Department of Medical Biochemistry; Dr. Ujong Peter U., Mr. Dasofunjo Kayode, Dr. Okafor
A.I, Prof. Edeogu, prof. Atangwho. J. I, Mr. Ati, Boniface, and all the staff of the Department.
God bless you all.

My utmost regard goes to my Parents; Mr./Mrs. Omini, Eyong Ikpi for their love and
immense support throughout the time of this seminar work. With all heart gratitude i say
thank you.

I won’t forget to acknowledge the love of my big brother, Brown, Eyong Ikpi and my friends
Andrew, Francis, Ezuma, Divine Ntui and others whose names might not appear here, I want to
say thank you all.
 SUMMARY

The primary function of the skin is to act as a barrier against insults from the environment, and
its unique structure reflects this. The skin is composed of two layers: the epidermal outer layer is
highly cellular and provides the barrier function, and the inner dermal layer ensures strength and
elasticity and gives nutritional support to the epidermis. Normal skin contains high
concentrations of vitamin C, which supports important and well-known functions, stimulating
collagen synthesis and assisting in antioxidant protection against UV-induced photo damage.
This knowledge is often used as a rationale for the addition of vitamin C to topical applications,
but the efficacy of such treatment, as opposed to optimizing dietary vitamin C intake, is poorly
understood. This review discusses the potential roles for vitamin C in skin health and
summarizes the in vitro and in vivo research to date. We compare the efficacy of nutritional
intake of vitamin C versus topical application, identify the areas where lack of evidence limits
our understanding of the potential benefits of vitamin C on skin health, and suggest which skin
properties are most likely to benefit from improved nutritional vitamin C intake.

Keywords: Ascorbate, dermis, epidermis, vitamin C, skin aging, collagen, UV protection, photo
damage.
 TABLE OF CONTENTS

Cover page - - - - - - - - - - i

Dedication - - - - - - - - - - ii

Acknowledgement - - - - - - - - - iii

Summary - - - - - - - - - - iv

Table of contents - - - - - - - - - v

CHAPTER ONE: INTRODUCTION

1.1 Background of study - - - - - - - - 1

CHAPTER TWO: GENERAL OVERVIEW OF VITAMINS

2.1 Vitamins - - - - - - - - - - 2

2.2 Classification of vitamins - - - - - - - - 2

2.2.1 Fat soluble vitamins - - - - - - - - 2

2.2.2 Water soluble - - - - - - - - - 3

2.3 Vitamin C - - - - - - - - - - 3

2.3.1 Dietary sources of Vitamin C - - - - - - - 5

2.3.2 Daily dietary recommendation for Vitamin C - - - - - 6

2.4 Functions of vitamin C - - - - - - - - 7

2.4.1 Role in immune system and inflammation - - - - - 7

2.4.2 Anti-oxidant property - - - - - - - - 8

2.4.3 Functions in cardiovascular diseases - - - - - - 9

2.4.4 Function of vitamin C in common cold - - - - - - 9

2.4.5 Vitamin C and bone formation - - - - - - - 10

2.5 Deficiencies of vitamin C - - - - - - - - 10

2.6 Side effects of Vitamin C - - - - - - - - 11

2.6.1 Oral route side effects - - - - - - - - 11

2.6.2 Parenteral route side effects - - - - - - - 12

CHAPTER THREE: METABOLISM

3.1 General metabolism - - - - - - - - 13

3.2 Absorption of vitamin C (Active transport) - - - - - 14

3.3 vitamin C distribution - - - - - - - - 15

3.4 Vitamin C excretion - - - - - - - - 15

3.5 Vitamin C storage - - - - - - - - - 16

CHAPTER FOUR: VITAMIN C AND THE SKIN

4.1 The role of vitamin C in the skin - - - - - - - 17

4.2 Bioavailability and uptake of Vitamin C in the skin - - - - 18

4.2.1 The Sodium-Dependent Vitamin C Transporters - - - - 18

4.3 Functions of vitamin C in the skin - - - - - - 19

4.3.1 The Promotion of Collagen Formation - - - - - - 19

4.3.2 The Ability to Scavenge Free Radicals and Dispose of Toxic Oxidants - 20

4.3.3 Inhibition of Melanogenesis - - - - - - - 20

4.3.4 Interaction with Cell Signalling Pathways - - - - - 20

4.3.5 Modulation of Epigenetic Pathways - - - - - - 21

4.4 Challenges to the Maintenance of Skin Health and Potential Protection by Vitamin C 22

4.4.1 Skin Aging - - - - - - - - - 22

4.4.2 Dry Skin - - - - - - - - - - 23

CHAPTER FIVE: CONCLUSION

5.1 Conclusion - - - - - - - - - 25

REFERENCES
 CHAPTER ONE

INTRODUCTION

1.2 Background of study

Vitamin C (Vit. C) Is one of the naturally occurring antioxidants in nature (Talakoub et al., 2009;
Traikovich 1999). Most plants and animals are able to synthesize Vitamin C in vivo from
glucose. Humans and certain other vertebrates lack the enzyme L-glucono-gamma lactone
oxidase required for in vivo synthesis of Vitamin C according to Farris (2009) hence, they must
acquire it from natural sources such as citrus fruits, green leafy vegetables, strawberries, papaya
and broccoli. The word “Ascorbus” means no Scurvy. Traditionally, Vitamin C rich foods like
lemons were carried by sailors on long journeys to avoid Scurvy, a disease of bleeding gums. In
1937, Dr. Albert Szent Goyrgi was awarded the Nobel Prize for his work in isolating the Vitamin
C molecule from red peppers and identifying its role in Scurvy (Wikepedia 2012).

L-ascorbic acid (LAA) is the chemically active form of Vitamin C. In nature, Vitamin C is found
in equal parts as LAA and D-ascorbic acid. These are essentially isomeric molecules and are
mutually interchangeable. However, only LAA is biologically active and thus useful in medical
practice (Talakoub et al., 2009). The absorption of Vitamin C in the gut is limited by an active
transport mechanism and hence a finite amount of the drug is absorbed despite high oral dosage.
Furthermore, bioavailability of Vitamin C in the skin is inadequate when it is administered
orally. The use of topical ascorbic acid is therefore favored in the practice of dermatology
(Matsuda et al., 2009).
 CHAPTER TWO

GENERAL OVERVIEW OF VITAMINS

2.1 Vitamins

The name “vitamin” comes from Casimir Funk, who in 1912 thought “vital amines” (similar to
amino acids) were responsible for preventing what we know now as vitamin deficiencies. He
coined the term “vitamines” to describe these organic substances that were recognized as
essential for life, yet unlike other organic nutrients (carbohydrates, protein, and fat), do not
provide energy to the body. Eventually, when scientists discovered that these compounds were
not amines, the ‘e’ was dropped to form the term “vitamins.”

Vitamins are essential, non-caloric, organic micronutrients. There is energy contained in the
chemical bonds of vitamin molecules, but our bodies don’t make the enzymes to break these
bonds and release their energy; instead, vitamins serve other essential functions in the body.

2.2 Classification of vitamins

2.2.1 Fat soluble vitamins

Vitamins can be classified as either a water-soluble or fat soluble vitamins. The fat-soluble
vitamins are very vital for the smooth functioning of the body. Their deficiencies have been
implicated in several health disorders. The recommended daily allowance (RDA) of the fat
soluble vitamins A, D, E, and K is 8000–1000 μg/day, 8000–5000 μg/day, 8–10 μg/day, and 70–
140 μg/day respectively (Kamangar & Emadi 2012). The RDA for water soluble vitamins
including thiamin, riboflavin, pyridoxine, niacin, biotin, ascorbic acid, and pantothenic acid are
1 mg/day, 1.2 mg/day, 2–2.2 mg/day, 13 mili-equivalents, 100–200 μg/day, 60 μg/day, and 4–
7 mg/day respectively (Kamangar & Emadi 2012).
2.2.2 Water soluble

The water-soluble vitamins consist of a mixed group of chemical compounds. Their

classification into specific chemical groups depends on both chemical characteristics and
functions.
The water-soluble vitamins consist of a mixed group of chemical compounds. Their
classification into specific chemical groups depends on both chemical characteristics and
functions. The letter designations (vitamins B1, B2, B3, etc., C) represent in part remnants from
the past as the discovery of given dietary growth or curative factors were given letter
designations. Vitamins are novel in their roles as “external” or dietary regulatory agents. They
have largely evolved to serve: 1) specific cofactor and/or co-substrate functions, 2) as regulatory
agents, or 3) as antioxidants. All of the vitamins undergo specific and metabolically controlled
modifications before activation or conversion into their functional forms. The most limiting
events that control function are often a specific step(s) in cofactor formation, e.g., a
phosphorylation reaction or ATP addition. For example, niacin, riboflavin, and ascorbic acid
serve primarily as redox cofactors. The roles of thiamin, pyridoxine (vitamin B-6) and
pantothenic acid (as a component of coenzyme A) are distinguished because of their importance
to carbohydrate, protein and amino acid, and acyl and acetyl transport, respectively. Folic acid,
vitamin B-12 (cobalamin), and biotin will be discussed in relationship to their roles in single-
carbon or CO2 transfer reactions.

2.3 Vitamin C

Vitamin C is a water-soluble vitamin that is necessary for normal growth and development. It is
an antioxidant that helps maintain the connective tissue protein collagen, protects against
infection, and helps iron absorption.

Vitamin C (ascorbic acid) is a water-soluble vitamin, which is necessary in the body to form
collagen in bones, cartilage, muscle, and blood vessels and aids in the absorption of iron. Dietary
sources of vitamin C include fruits and vegetables, particularly citrus fruits such as oranges.
Severe deficiency of vitamin C causes scurvy. Although rare, scurvy includes potentially severe
consequences, and can cause sudden death. Patients with scurvy are treated with vitamin C and
should be under medical supervision. Many uses for vitamin C have been proposed, but few have
been found to be beneficial in scientific studies. In particular, research in asthma, cancer, and
diabetes remains inconclusive, and no benefits have been found in the prevention of cataracts or
heart disease.

Although vitamin C is the generic name of l-ascorbic acid, it has many other chemical names as
ascorbate and antiscorbutic vitamin. L-Ascorbic acid molecule is formed of asymmetrical six-
carbon atoms (C6H8O6) which is structurally related to glucose (Velisek & Cejpek 2007). Its
molecular weight is 176 with a melting point of 190–192°C (with decomposition) and shows a
density of approximately 1.65 g/cm3. L-Ascorbic acid is freely soluble in water (300 g/L at
20°C), difficult in alcohol (20 g/L at 20°C) and insoluble in chloroform, ether and benzene. It
forms a clear colorless to slightly yellow solution. It has two pKa values: 4.2 and 11.6. The pH of
a 5% (w/v) solution in water is 2.2–2.5. The chemical structure of ascorbic acid determines its
physical and chemical properties. It is a weak, water soluble, unstable organic acid which can be
easily oxidized or destroyed in light, aerobic condition (oxygen), high temperature, alkali,
humidity, copper and heavy metals. Ascorbic acid is usually found in the form of white or
slightly yellowish crystalline powder. Its crystalline form is chemically stable in dryness.
However L-ascorbic acid is highly soluble in water, it shows great difficulty to be soluble in
alcohol, chloroform, ether and benzene. In water, it forms clear colorless slightly yellow solution
which is rapidly oxidized (Calder et al., 2009). There are many derivatives of ascorbic acid as
sodium l-ascorbate (sodium ascorbate), calcium l-ascorbate (calcium ascorbate), zinc-ascorbate,
6-palmityll-ascorbic acid (ascorbyl palmitate) and ascorbyl monophosphate calcium sodium salt
(sodium calcium ascorbyl phosphate).

Ascorbic acid is obtained from sodium ascorbate by cation exchange. While sodium ascorbate
results from reacting methyl-d-sorbosonate (or ketogulonic acid methyl ester) with sodium
carbonate. Calcium ascorbate is produced by the interaction of ascorbic acid with calcium
carbonate in water and ethanol, which it is then isolated and dried. Ascorbyl palmitate is
prepared by reaction of ascorbic acid with sulfuric acid followed by esterification with palmitic
acid. Sodium calcium ascorbyl phosphate resulted from the reaction of ascorbic acid (alone or in
combination with sodium ascorbate) with calcium hydroxide and sodium trimetaphosphate. The
previous ascorbic acid derivatives have superior properties in comparison to ascorbic acid as the
light resistance, skin irritation.
 Plate 2.1: structure of vitamin C (Acorbic acid)

2.3.1 Dietary sources of Vitamin C

Vitamin C is produced only in non-humans as primate species, guinea pigs, fishes and birds
(Kleszczewska et al., 2000). Although most of the animals have the ability to synthesis their
needs of vitamin C, humans suffer from mutation in the DNA coding of gulonolactone oxidase
which is the main enzyme responsible for ascorbic acid synthesis (Nishikimi et al., 1994). Due to
this mutation, the external supplement of vitamin C becomes a must (Garriguet 2010). The main
source of vitamin C for human beings is mainly found in fruits and vegetables. Citrus fruits and
other types are particularly rich sources of vitamin C as; cantaloupe, water melon, berries,
pineapple, strawberries, cherries, kiwi fruits, mangoes, and tomatoes. Furthermore, vegetables
are considered the main source of vitamin C due to its higher content and availability for longer
period throughout the year such as cabbage, broccoli, Brussels sprouts, bean sprouts, cauliflower,
mustard greens, peppers, peas and potatoes (Haytowitz 1995).
 Plate 2.2: Amount of vitamin C in different sources

2.3.2 Daily dietary recommendation for Vitamin C

Table 2.1: dietary intake of vitamin C

2.4 Functions of vitamin C

Vitamin C plays an important role in many physiological processes in humans. It is needed for
the repair of tissues in all parts of the body. The important functions of vitamin C include the
formation of protein used to make skin, tendons, ligaments, and blood vessels for healing
wounds and forming scar tissue, for repairing and maintaining cartilage, bones, and teeth and aid
in the absorption of iron. It can also act as a reducing and capping agent for metal nanoparticles.

Plate 2.3: Sources and multifunctional role of vitamin C in metabolic processes in human body

Role in immune system and inflammation

Vitamin C has an important role in the maintenance of a healthy immune system and its
deficiency causes immune insufficiency and multiple infections. The ascorbic acid level is
lowered in various body fluids during bacterial infections. Thus, it is commonly used as
adjunctive treatment in many infectious diseases such as hepatitis, HIV, influenza and
periodontal diseases (Kalokerinos et al., 2005).

Vitamin C administration modifies and enhances both the innate and adaptive immune response.
It neutralizes the bacterial toxins especially endotoxins by blocking the essential signal for
lipopolysaccharides (LPS) formation. On the other hand, LPS block the passage of ascorbic acid
through blood brain barrier and inhibits its uptake by various cells (Kalokerinos et al., 2005).
Ascorbic acid improves the phagocytic properties and activity of various immune cells including
neutrophils, natural killer cells, macrophages and lymphocytes. Vitamin C increases
lymphocytes proliferation and antibody production.

Anti-oxidant property

Oxidative stress/ROS have a main role in inflammatory diseases including periodontal diseases
(Weidinger & Kozlov 2015). The ROS are classified into 3 classes; the first are reactive free
radicals as oxygen related radicals (superoxide, hydroxyl radical or peroxyl radicals). The second
class is reactive species but not free radicals as hypochlorous acid. The third class is radicals
resulted from the reaction with ascorbic acid (Neuzil et al., 1997). Antioxidants are also
classified into enzymatic and non-enzymatic. The enzymatic antioxidants include catalase
enzyme, thiol-containing agents (cysteine, methionine, taurine), glutathione and lipoic acid
(Mansour et al., 2002). Vitamin C is one of the nutrient non-enzymatic anti-oxidants. Its
antioxidant effect is by electron donation process where vitamin C easily donates two electrons
(reduction reaction) to other compounds in order to prevent its oxidation. When ascorbic acid
donates the first electron, it is transformed into a free radical called ascorbyl radical (semi-
dehydroascorbic acid). It is a relatively stable, unreactive free radical with unbound electron in
its outer shell but it has a short life time (10–15 s). The unreactivity of this radical makes it
unharmful to the surrounding cells. This process is called free radical scavenging or quenching.
When it donates the second electron, it transformed into dehydroascorbic acid. Its stability may
only last for few minutes. As a general rule, it was detected that vitamin C acts as a pro-oxidant
at low doses and acts as an antioxidant in high doses. It was also detected that the level of
vitamin C in the skin usually exposed to ultraviolet radiation is lower than that exposed lesser.
The antioxidant activity of vitamin C enhances the epidermal turn over, and the movement of
young cells to the surface of the skin where they replace old cells. The study conducted by Frank
in (2000) showed that RNA improved the ability of the skin cells to utilize oxygen. Ascorbyl
radical and dehydroascorbic acid are reversible agents which can easily rebound into ascorbic
acid. These reversible agents can irreversibly transformed into 2,3-diketogulonic acid which is
further metabolized into xylose, xylonate, lyxonate and oxalate. Vitamin C is considered as a
strong anti-inflammatory agent as it inhibits many types of inflammatory mediators as tumor
necrosis factor alpha.

This property is commonly used in the treatment of postoperative erythema formed after CO2
laser in skin resurfacing. In 1987, Halliwell detected significant reduction of plasma levels of
ascorbic acid in association with elevated histamine in inflammatory diseases as ulcerative colitis
and rheumatoid arthritis. This was explained by the discovery of the anti-histaminic effect of
vitamin C. It was also found that the higher ascorbic acid content in joints, the higher protection
levels against damage which directed many physicians to use ascorbic acid in combination
therapy with drugs aiming to joint protection as glucosamine. It was discovered that vitamin C
has an efficient chemotherapeutic effect. The cytotoxic effect of vitamin C is dose and route
dependent. The tumor cells are more sensitive to high intravenous (cytotoxic) levels of vitamin C
than the normal ones. At the administration of 10 g of intravenous vitamin C, a marked elevation
of the extracellular concentration (1000 μmol/L) is detected which have a toxic effect on the
cancer cells due to the action of the ascorbyl radicals (free radical species) (] Sakagami et al.,
2000). On the contrary to the cancer cells, normal cells can compensate the damage occurred by
these oxidative species. It was also found that these mega doses of vitamin C are essential in
other diseases as diabetes, cataracts, glaucoma, macular degeneration, atherosclerosis, stroke and
heart diseases (Will et al., 1996).

Functions in cardiovascular diseases

The antioxidant property of vitamin C helps for the treatment of cardiovascular diseases. Vitamin
C has the capability for reducing monocyte adherence to the endothelium, improving
endothelium-dependent nitric oxide production and vasodilation and reducing vascular smooth-
muscle-cell apoptosis, which prevents plaque instability in atherosclerosis (Honarbakhsh &
Schachter 2008). The oxidative damage including the oxidative modification of low-density
lipoproteins is a major cause of cardiovascular disease. The antioxidant property of vitamin C
helps to reduce this to a certain extent (Willcox et al., 2008).

Function of vitamin C in common cold

Pauling in 1970 suggested that vitamin C can be used for the treatment of common cold (Pauling
1971). There are so many reports in Cochrane Database Syst. Review showing the use of
prophylactic vitamin C reduces the cold duration in adults and children (Douglas et al., 2007).
The use of vitamin C might reduce the duration of common cold due to its anti-histamine effect
of high dose of vitamin C.

However, the results are inconsistent and still research is undergoing in this field. There are
Database Syst. Review showing the use of prophylactic vitamin C reduces the cold duration in
adults and children. The use of vitamin C might reduce the duration of common cold due to its
anti-histamine effect of high dose of vitamin C.

Vitamin C and bone formation

As previously mentioned, vitamin C increases the production of collagen type I and X needed for
matrix formation, activation of osteoblast growth and differentiation (Yilmaz et al., 2001). It is
also needed in order to maintain adequate bone density. Stimulation and higher expression of
osteocalcin and osteonectin on the osteoblasts was also reported. In postmenopausal women,
higher levels of vitamin C are needed in order to protect against bone abnormalities as it is
considered as delaying osteoporosis factor. Furthermore, in scurvy, lower bone density with
marked bone abnormalities commonly reported (Otsuka et al., 2000). In case of deficiency
occurs in young individuals, bone fragility, cartilage resorption and fracture of growth plates.
The detected abnormalities were attributed to reduced activity of osteocytes and chondrocytes. It
also maintains and preserves the balance between osteoblasts and osteoclasts. In order to achieve
optimum proliferation of the osteoblasts and fibroblasts, 200 μg/ml is the maximum dose needed.
Apoptosis occurs when exceeding such dose. In 2004, an in vitro study used vitamin C with
scaffolds in tissue engineering in order to regenerate bone. The sustained released vitamin C
stimulates the formation of type I collagen and alkaline phosphatase (Zhang et al., 2003). In
2013, Fu et al., (2013) used isotonic irrigation of ascorbic acid derivative during grafting of the
anterior tendon of the knee joint. Significant reduction of the inflammatory response in the
surgical site due to lack of toxicity, irritation, watery consistency and potent anti-oxidant effect.

2.5 Deficiencies of vitamin C

In scurvy, absence of wound healing and failure of fractured bones to heal. This was explained
by the deficiency of collagen formation due to the vitamin C deficiency. Scurvy could be
produced if reduction of the body reservoir of vitamin C into its fifth. The required body
reservoir and the needed dosage are determinate according to the body weight (Rathee 2013).

In scurvy, body weakness, legs and arms edema, nose, skin and gums hemorrhage, infections,
bone and cartilage damage (osteoporosis), vasculitis and cardiomegaly. Many forms of bleeding
found as petechiae, subperiosteal hemorrhage, ecchymoses, purpura, bleeding gums,
hemarthrosis.

2.6 Side effects of Vitamin C

Oral route side effects

Side effects of vitamin C could be only detected with large doses exceeding the ULs for each
individual especially on a single intake. Most of the vitamin C drawbacks were reported during
oral uptake. They include diarrhea, abdominal pain, renal stones and enamel erosion during
chewing.

GIT disturbance

Because of the poor oral bioavailability of vitamin C, toxic signs and symptoms may appear with
large doses as a single dose. Diarrhea can occur. To avoid diarrhea occurrence, 2 g is the
maximum permissible single dose (National Academy of Sciences, 2000). Diarrhea and
abdominal pain may occur due to the excretion of large amount of un-metabolized vitamin C.
Such manifestations are dose related. It can be controlled by either, reducing the total daily dose,
dividing the total dose into multiple small doses, administrating the vitamin with food to
decrease its absorption or to take the buffered form of vitamin C as sodium ascorbate or calcium
ascorbate (National Academy of Sciences, 2000). Even the usage of encapsulated vitamin C
could not protect against the gastric upset. The gastrointestinal symptoms usually disappear
within 1–2 weeks.

Renal stones

Vitamin C metabolism results in calcium oxalate salts. Formation of renal stones (oxalate salts)
and oxaluria are resulted in overdoses of vitamin C (Urivetsky et al., 1992). Later on, it was
detected that this oxaluria is usually due to laboratory artifact occurs in the urine collection tube
(ex vivo). It was also detected that vitamin C counteracting the formation of calcium oxalate
crystals because of its ability to bind to calcium found in urine. Vitamin C also has the ability to
increase the solubility of the calcium oxalate due to its mild acidity. It also triggers the normal
urination process and prevents urine retention. All the previous actions decrease the incidence of
kidney stones formation. Also, it was found that vitamin C increases the urinary execration of
uric acid and decreasing the plasma level of uric acid. But others demand the uricosuric effects
of vitamin C as the rapid migration of uric acid from tissues. It was detected that 1 or 2 g per day
increases the urinary oxalate stones were detected. Some studies detect lowering of urine pH
after vitamin C intake. The renal stones incidence of accumulation occurs in an average
concentration of 60 g (IV) and more than 5 g (oral) (Mashour et al., 2000). In case of renal
insufficiency, 1 g/day for 3 months is enough to produce renal stones (Alkhunaizi and Chan,
1996).

Metabolism side effects

It accelerates the absorption of other heavy metals as lead and mercury which increases its
toxicity (Wyngaarded, 1987). In patients with high iron stores, vitamin C worsens the state. It
also increases the iron-induced oxidative damage (Slivaka et al., 1986).

Parenteral route side effects

During injection, transited mild soreness occurs during intramuscular, subcutaneous routes.
Faintness or dizziness was reported on rapid intravenous administration. The renal stones
incidence of accumulation occurs in an average concentration of 60 g (IV) and more than 5 g
(oral) [163]. In case of renal insufficiency, 1 g/day for 3 months is enough to produce renal
stones (Alkhunaizi and Chan, 1996).
 CHAPTER THREE

METABOLISM

3.1 General metabolism

Vitamin C functions depend mainly on its main character as a reducing agent and the results of
its oxidation mechanisms either reversible or irreversible. These reactions depend only on the pH
changes and not on the presence of air or oxidizing agents.

Ascorbic acid undergoes a 3-step oxidation process. In the beginning, ascorbic acid can
reversibly oxidize into dehydroascorbic acid on the exposure to copper, low alkaline media and
heat (Thurnham 2000). Dehydroascorbic acid is a very short half-life (few minutes) product
which can either reversibly or irreversibly oxidize in the tissues. In pH 4.0, ordinary
temperatures and aqueous media, dehydroascorbic acid can be oxidized irreversibly into 2,3-
diketol-glutonic acid (diketogulonic acid). However, the dehydroascorbic acid oxidation begins
in mild acidic media (pH 4.0), it requires a neutral or alkaline media to progress more rapidly.
The resultant diketogulonic acid is a stronger reducing agent, not reduced by glutathione or H2S
and not an anti-ascorbutic agent. It was found that below pH 4.0, diketogulonic acid losses its
reducing property. In acidic media and the presence of H2S, dehydroascorbic acid can also
reversibly change into ascorbic acid. Ascorbic acid and dehydroascorbic acid have the same anti-
ascorbutic effect. The third oxidation product is l-threonic acid and oxalic acid which proceed
only in alkaline media (pH 7–9) (Thurnham 2000). All reversible changes can be done in the
presence of H2S and glutathione in neutral or alkaline media. Sometimes, carbon dioxide may be
the result of vitamin C oxidation at high doses. In human beings, ascorbic acid is reversibly
oxidized into dehydroascorbic acid, which can be reduced back to ascorbic acid or hydrolyzed to
diketogulonic acid and then oxidized into oxalic acid, threonic acid, xylose, xylonic acid and
lyxonic acid. Further oxidation (decomposition) may occur by the oxidizing agents in food.
According to the oxidation-reduction reactions, ascorbic acid is the reduced form of vitamin C
while dehydroascorbic acid is the oxidized form of vitamin C. The l-isomer of ascorbic acid is
the only active form. Other isomers as d-ascorbic acid, d-isoascorbic acid and l-isoascorbic acid
are present. These stereoisomers have no effect in the treatment of scurvy. The absorbed and the
unabsorbed forms of ascorbic acid can be excreted in conjugated or non-conjugated pattern.
Ascorbic acid may undergo limited conjugation with sulfate to form ascorbate-2-sulphate, which
is excreted in the urine. Unchanged ascorbic acid and its metabolites are excreted in the urine. In
the presence of intestinal flora, high doses of ascorbic acid (unabsorbed part) can oxidized into
carbon dioxide which is the main excretory mechanism of vitamin C in guinea pigs, rats and
rabbits. There exists equilibrium between ascorbic acid and dehydro-ascorbic acid, dependent on
the redox status of the cells.

Plate 3.1: metabolism of ascorbic acid

3.2 Absorption of vitamin C (Active transport)

The hydrophilic nature of ascorbic acid facilitates its absorption through buccal mucosa, stomach
and small intestine. Its absorption depends mainly on passive diffusion through the buccal
mucosa (Stevenson 1974). Vitamin C absorption occurs through small intestine (distal intestine)
by active transport mechanism. Sodium electrochemical gradient is the process by which active
transport of ascorbic acid occurs. This process proceeds by the help of sodium vitamin C
transporter type 1 (SVCT1). This is the same transporter responsible for vitamin C transport in
retina. SVCT2 is responsible for transporting vitamin C into brain, lung, liver, heart and skeletal
muscles. The absorption process is usually inhibited by glucose.

The majority of ascorbate is transported by SVCT1 in epithelial cells (e.g., intestine, kidney and
liver), and the remaining is transported by SVCT2 in specialized cells (e.g., brain and eye). The
main concentrations of vitamin C are located in brain and adrenal cells. The oxidative products
of vitamin C as dehydroascorbic acid are transported faster into cells than the pure form. While
the absorption of low doses (15–30 mg) is very high (up to 98%), ascorbic acid absorption
decreases (50%) with larger doses (1000–1250 mg) which is commonly administrated in acute
illness. In human blood, ascorbic acid is always found in the reduced form (ascorbic acid). It was
also found that the red blood corpuscles are not permeable for ascorbic acid and also to glucose.
It oxidized very slowly in blood than in plasma (no oxidation reactions occur). Its normal plasma
level ranges between 50 and 100 μM according to the diet intake in healthy non-smoker
individuals. Increasing the plasma level and the intracellular level is not a dose dependent. Its
intracellular level is higher than the plasma level. The plasma level does not increase above the
normal range even by increasing the intake into 500 mg because of its excellent excretion from
kidneys through urine.

3.3 vitamin C distribution

Vitamin C is widely distributed in all the body tissues. Its level is high in adrenal gland, pituitary
gland, and retina. Its level decreases in kidneys and muscles.

3.4 Vitamin C excretion

Vitamin C metabolites (oxalate salts) and unmetabolized vitamin C are excreted by kidneys. Few
percentage of vitamin C is excreted through feces. The urinary excretion of vitamin C is dose
dependent. Less than 100 mg/day, no vitamin C was detected in urine. At 100 mg/day, 25% of its
amount was excreted in urine. The latter percentage is doubled with the administration of
200 mg/day. At high doses, large amount of unmetabolized vitamin C is excreted. The higher
doses of vitamin C intake, the higher vitamin C concentration in blood and tissues occurs. As a
response for high doses, vitamin C excretion from kidneys and sweat occur. The antiviral and
anti-bacterial effect of vitamin C protects skin and kidneys from infection (Pauling 1970). Also
in extra doses, the oxidation components were used as an anticancer effect more the vitamin C
itself. It was found that the excretion of ascorbic acid when administrating 400 mg ascorbic acid
ranges between 30 and 50% in healthy individuals. This percent decreases in diseased patients
due to higher consumption. Repeated low doses (about 200 mg) are highly recommended in
diseased individuals due to theses low doses saturate the body. Extremely low dosages
(90 mg/day) could result in inability of the immune system to respond to diseases as
degenerative diseases. Therefore, limited renal clearance of ascorbic acid is usually detected. The
plasma saturation of ascorbic acid at 70 μM (0.123 mg/dl). This level controls the excretion of
the ascorbic acid through kidneys. At Higher plasma levels (above 70 μM), higher excretion
levels are usually detected. The intravenous route exerts 30–70 folds of vitamin C plasma levels
than the oral route. The rapid excretion due to its water soluble nature limits its harmful effect
and makes it totally safe product in normal doses. It also found that the upper tolerable limit
(UL) is 2 g. Depending on the depletion-repletion study; it was found that the RDA is 75 mg for
women and 90 mg for men. It was modified by Levine et al. in 2001 into the administration of
90 mg to both sexes. The maximum bioavailability and absorption of vitamin C achieved at
500 mg (Pacier & Martirosyan 2015).

3.5 Vitamin C storage

In 1936, Marinesco et al., detected the lower levels of ascorbic acid in others organs as pancreas,
spleen and thymus gland. Plaut and Billow detected the ascorbic acid lowering not only in the
organs but also in body fluids as CSF, blood and urine. They also detected this deficiency in
neural diseases and alcoholism. Many reasons were thought to be the cause of vitamin C
deficiency in old people. Decreased intestinal absorption and dietary deficiency are the main
causes. In human blood, ascorbic acid is always found in the reduced form (ascorbic acid). It was
also found that the red blood corpuscles are not permeable for ascorbic acid and also to glucose.
It oxidized very slowly in blood than in plasma (no oxidation reactions occur). Its normal plasma
level ranges between 50 and 100 μM according to the diet intake in healthy non-smoker
individuals. Increasing the plasma level and the intracellular level is not a dose dependent. Its
intracellular level is higher than the plasma level. The plasma level does not increase above the
normal range even by increasing the intake into 500 mg because of its excellent excretion from
kidneys through urine. In 1934, Yavorsky et al., (1934) analyzed the ascorbic acid amount found
in the different body organs at different ages.
 CHAPTER FOUR

VITAMIN C AND THE SKIN

4.1 The role of vitamin C in the skin

The skin is a multi-functional organ, the largest in the body, and its appearance generally reflects
the health and efficacy of its underlying structures. It has many functions, but its fundamental
role is to provide a protective interface between the external environment and an individual’s
tissues, providing shielding from mechanical and chemical threats, pathogens, ultraviolet
radiation and even dehydration (Weller et al., 2008). Being in constant contact with the external
environment, the skin is subject to more insults than most of our other organs, and is where the
first visible signs of aging occur.

It is accepted that the nutritional status of with respect to both macronutrients and micronutrients
is important for skin health and appearance. Evidence of this is provided by the many vitamin
deficiency diseases that result in significant disorders of the skin (Boelsma et al., 2003).
Dermatological signs of B vitamin deficiency, for example, include a patchy red rash,
seborrhoeic dermatitis and fungal skin and nail infections. The vitamin C deficiency disease
scurvy is characterized by skin fragility, bleeding gums and corkscrew hairs as well as impaired
wound healing. Nutritional status is vital for maintaining normal functioning of the skin during
collagen synthesis and keratinocyte differentiation. Additionally, many of the components of our
antioxidant defenses such as vitamins C and E and selenium are obtained from the diet, and these
are likely to be important for protection against UV induced damages.

Normal skin contains high concentrations of vitamin C with levels comparable to other body
tissues and well above plasma concentrations, suggesting active accumulation from the
circulation. Most of the vitamin C in the skin appears to be in intracellular compartments, with
concentrations likely to be in the millimolar range [25–27]. It is transported into cells from the
blood vessels present in the dermal layer. Skin vitamin C levels have not often been reported and
there is considerable variation in the published levels, with a 10-fold range across a number of
independent studies. Levels are similar to that found in numerous other body organs. The
variation in reported levels most likely reflects the difficulty in handling skin tissue, which is
very resilient to degradation and solubilisation, but may also be due to the location of the skin
sample and the age of the donor.

Several reports have indicated that vitamin C levels are lower in aged or photo-damaged skin.
Whether this association reflects cause or effect is unknown, but it has also been reported that
excessive exposure to oxidant stress via pollutants or UV irradiation is associated with depleted
vitamin C levels in the epidermal layer (Shindo et al., 1994). Indeed, more vitamin C is found in
the epidermal layer than in the dermis, with differences of 2–5-fold between the two layers being
consistently reported (Rhie et al., 2001). Levels of vitamin C in skin are similar to the levels of
other water soluble antioxidants such as glutathione. There is a suggestion that vitamin C in the
stratum corneum layer of the epidermis exists in a concentration gradient. The lowest vitamin C
concentration was present at the outer surface of the epidermis of the SKH-1 hairless mouse, a
model of human skin, with a sharp increase in concentration in the deeper layers of the stratum
corneum, possibly reflecting depletion in the outer cells due to chronic exposure to the
environment.

4.2 Bioavailability and uptake of Vitamin C in the skin

4.2.1 The Sodium-Dependent Vitamin C Transporters

Vitamin C uptake from the plasma and transport across the skin layers is mediated by specific
sodium-dependent vitamin C transporters (SVCTs) that are present throughout the body and are
also responsible for transport into other tissues. Interestingly, cells in the epidermis express both
types of vitamin C transporter, SVCT1 and SVCT2 (Steiling et al., 2007). This contrasts with
most other tissues, which express SVCT2 only. SVCT1 expression in the body is largely
confined to the epithelial cells in the small intestine and the kidney and is associated with active
inter-cellular transport of the vitamin. The specific localization of SVCT1 in the epidermis is of
interest due to the lack of vasculature in this tissue, and suggests that the combined expression of
both transporters 1 and 2 ensures effective uptake and intracellular accumulation of the vitamin.
Together with the high levels of vitamin C measured in the epidermal layer, the dual expression
of the SVCTs suggests a high dependency on vitamin C in this tissue (May 2011). Both
transporters are hydrophobic membrane proteins that co-transport sodium, driving the uptake of
vitamin C into cells. Replacement of sodium with other positively charged ions completely
abolishes transport. SVCT1 and SVCT2 have quite different uptake kinetics reflecting their
different physiological functions. SVCT1 transports vitamin C with a low affinity but with a high
capacity (Km of 65–237 µmol/L) mediating uptake of vitamin C from the diet and re-uptake in
the tubule cells in the kidney (Savini et al., 2008). SVCT2, which is present in almost every cell
in the body, is thought to be a high-affinity, low capacity transporter, with a Km of ~20 µM
meaning it can function at low concentrations of vitamin C (Savini et al., 2008). As well as
transporter affinity, vitamin C transport is regulated by the availability of the SVCT proteins on
the plasma membrane.

4.3 Functions of vitamin C in the skin

The high concentration of vitamin C in the skin indicates that it has a number of important
biological functions that are relevant to skin health. Based on what we know about vitamin C
function, attention has been focused on collagen formation and antioxidant protection; however,
evidence is emerging for other activities.

The Promotion of Collagen Formation

Vitamin C acts as a co-factor for the proline and lysine hydroxylases that stabilize the collagen
molecule tertiary structure, and it also promotes collagen gene expression. In the skin, collagen
formation is carried out mostly by the fibroblasts in the dermis, resulting in the generation of the
basement membrane and dermal collagen matrix. The dependence of the collagen hydroxylase
enzymes on vitamin C has been demonstrated in a number of studies with fibroblast cells in vitro
(Hinek et al., 2014), with both decreased total synthesis and decreased cross-linking when
vitamin C is absent. The activity of the hydroxylases is much more difficult to measure in vivo,
as the amount of collagen synthesized may vary only a little. Rather, animal studies with the
vitamin-C-deficient GULO mouse indicate that the stability of the synthesized collagen varies
with vitamin C availability, reflecting the stabilizing function of the collagen cross-links formed
by the hydroxylases. In addition to stabilizing the collagen molecule by hydroxylation, vitamin C
also stimulates collagen mRNA production by fibroblasts.
The Ability to Scavenge Free Radicals and Dispose of Toxic Oxidants

Vitamin C is a potent antioxidant that can neutralize and remove oxidants, such as those found in
environmental pollutants and after exposure to ultraviolet radiation. This activity appears to be of
particular importance in the epidermis, where vitamin C is concentrated in the skin. However,
vitamin C is only one player in the antioxidant arsenal that includes enzymatic defenses
(catalase, glutathione peroxidase and superoxide dismutase) as well as other non-enzymatic
defenses (vitamin E, glutathione, uric acid and other putative antioxidants such as carotenoids)
(Agrawal et al., 2014). Most intervention studies carried out to determine the capacity of
antioxidants to prevent oxidative damage to skin have used a cocktail of these compounds.
Vitamin C is particularly effective at reducing oxidative damage to the skin when it is used in
conjunction with vitamin E. This is in accord with its known function as a regenerator of
oxidized vitamin E, thereby effectively recycling this important lipid-soluble radical scavenger
and limiting oxidative damage to cell membrane structures.

Inhibition of Melanogenesis

Vitamin C derivatives, including the magnesium phosophate ascorbyl derivative, have been
shown to decrease melanin synthesis both in cultured melanocytes and in vivo. This activity has
been proposed to be due to its ability to interfere with the action of tyrosinase, the rate-limiting
enzyme in melanogenesis. Tyrosinase catalyses the hydroxylation of tyrosine to
dihydroxyphenylalanine (DOPA), and the oxidation of DOPA to its corresponding ortho-
quinone. The inhibition in melanin production by vitamin C is thought to be due to the vitamin’s
ability to reduce the ortho-quinones generated by tyrosinase, although other mechanisms are also
possible (Ebanks et al., 2009). Agents that decrease melanogenesis are used to treat skin hyper-
pigmentation in conditions such as melisma or age spots.

Interaction with Cell Signalling Pathways

In vitro studies clearly show that vitamin C can play a role in the differentiation of keratinocytes.
For example, vitamin C enhanced the differentiation of rat epidermal keratinocytes cells in an
organotypic culture model, with markedly improved ultrastructural organization of the stratum
corneum, accompanied by enhanced barrier function. Vitamin C also increased numbers of
keratohyalin granules and levels of the late differentiation marker filaggrin, which appeared to be
due to altered gene expression. Others have also shown that vitamin C promotes synthesis and
organization of barrier lipids and increased cornfield envelope formation during differentiation.
The mechanism(s) by which vitamin C modulates keratinocyte differentiation is not yet
elucidated; however, it has been hypothesized to be under the control of protein kinase C and
AP-1.

In addition to vitamin C’s ability to promote collagen synthesis, there is evidence to suggest that
vitamin C increases proliferation and migration of dermal fibroblasts, functions vital for effective
wound healing, although the underlying mechanisms driving this activity are not yet known
(Duarte et al., 2009). Through the stimulation of regulatory hydroxylases, vitamin C also
regulates the stabilization and activation of the hypoxia-inducible factor (HIF)-1, a metabolic
sensor that controls the expression of hundreds of genes involved with cell survival and tissue
remodeling, including collagenases. Vitamin C has been shown to both stimulate and inhibit
elastin synthesis in cultured fibroblasts (Davidson et al., 1997). Glycosaminoglycan synthesis as
part of extracellular matrix formation is also increased by vitamin C treatment, and it may also
influence gene expression of antioxidant enzymes, including those involved in DNA repair. As
such, vitamin C has been shown to increase the repair of oxidatively damaged bases. The
modulation of gene expression may be important for its ability to protect during UV exposure via
its inhibition of pro-inflammatory cytokine secretion and apoptosis.

Modulation of Epigenetic Pathways

In addition to the gene regulatory activities listed above, vitamin C has a role in epigenetic
regulation of gene expression by functioning as a co-factor for the ten-eleven translocation
(TET) family of enzymes, which catalyse the removal of methylated cytosine through its
hydroxylation to 5-hydroxymethylcytosine (5 hmC). As well as being a DNA demethylation
intermediate, it appears that 5 hmC is an epigenetic mark in its own right, with transcriptional
regulatory activity (Song & He 2013). Aberrant epigenetic alterations are thought to have a role
in cancer progression, and there is data to suggest that a loss of 5 hmC occurs during the early
development and progression of melanoma. Interestingly, vitamin C treatment has been shown to
increase 5 hmC content in melanoma cell lines, also causing a consequent alteration in the
transcriptome and a decrease in malignant phenotype (Gustafson et al., 2015). Because TETs
have a specific requirement for vitamin C to maintain enzyme activity, this provides a further
mechanism by which the vitamin may affect gene expression and cell function. For example, Lin
and co-workers showed that vitamin C protected against UV-induced apoptosis of an epidermal
cell line via a TET-dependent mechanism, which involved increases in p21 and p16 gene
expression (Lin et al., 2014).

4.3 Challenges to the Maintenance of Skin Health and Potential Protection by Vitamin C

During the course of a normal lifetime, the skin is exposed to a number of challenges that can
affect structure, function and appearance, including:

• Deterioration due to normal aging, contributing to loss of elasticity and wrinkle formation.

• Exposure to the elements, leading to discoloration, dryness and accelerated wrinkling.

• Chemical insults including exposure to oxidizing beauty and cleansing products (hair dyes,
soaps, detergents, bleaches).

• Direct injury, as in wounding and burning.

Vitamin C may provide significant protection against these changes and regeneration of healthy
skin following insult and injury is a goal for most of us. The following sections review the
available evidence of a role for vitamin C in the maintenance of healthy skin and the prevention
of damage.

Skin Aging

Like the rest of the human body, the skin is subject to changes caused by the process of natural
aging. All skin layers show age-related changes in structure and functional capacity (Blume-
Peytavi et al., 2016) and, as occurs in other body systems, this may result in increased
susceptibility to a variety of disorders and diseases, such as the development of dermatoses and
skin cancer (Blume-Peytavi et al., 2016). As well as this, changes in the appearance of skin are
often the first visible signs of aging and this can have implications for our emotional and mental
wellbeing. Aging of skin can be thought of as two distinct processes—natural or ‘intrinsic’
aging, caused simply by the passage of time, and environmental aging. Lifestyle factors such as
smoking and exposure to environmental pollutants increase the rate of environmental aging, and
can have a marked impact on the function and appearance of skin. Exposure to chronic
ultraviolet radiation from sunlight is also a major environmental factor that prematurely damages
our skin. The changes due to environmental aging are usually superimposed on those that occur
naturally, often making it difficult to distinguish between the two.

The ability of vitamin C to limit natural aging is difficult to distinguish from its ability to prevent
the additional insults due to excessive sun exposure, smoking or environmental stress and there
is very limited information available concerning a relationship between vitamin C levels and
general skin deterioration. The most compelling argument for a role of vitamin C in protecting
skin function comes from observations that deficiency causes obvious skin problems—early
signs of scurvy, for example, include skin fragility, corkscrew hairs and poor wound healing
(Ellinger & Stehle 2009).

Because vitamin C deficiency results in impaired function, it is assumed that increasing intake
will be beneficial. However, there are no studies that have measured vitamin C levels or intake
and associated aging changes (Rinnerthaler et al., 2015). Vitamin C is almost never measured in
the skin and this information is needed before we can improve our understanding of what level of
intake might be beneficial for skin health and protection against aging-related changes.

Dry Skin

Dry skin is a common condition typically experienced by most people at some stage in their
lives. It can occur in response to a particular skin care regime, illness, medications, or due to
environmental changes in temperature, air flow and humidity. The prevalence of dry skin also
increases with age; this was originally believed to be due to decreased water content or sebum
production in the skin as we get older. However, it is now considered likely to be due to
alterations in the keratinisation process and lipid content of the stratum corneum (White-Chu &
Reddy 2011).

The pathogenesis of dry skin is becoming clearer and three contributing deficiencies have been
identified.

• A deficiency in the skin barrier lipids, the ceramides, has been identified. These lipids are the
main intercellular lipids in the stratum corneum, accounting for 40 to 50 percent of total lipids.
• A reduction in substances known as the natural moisturizing factor (NMF) is also thought to be
involved in dry skin. These substances are found in the stratum corneum within the corneocytes,
where they bind water, allowing the corneocyte to remain hydrated despite the drying effects of
the environment.

• More recently, a deficiency of the skin’s own moisture network in the epidermis, mediated by
the newly discovered aquaporin water channels, has been suggested to play a role.

Cell culture studies have shown that the addition of vitamin C enhances the production of barrier
lipids and induces differentiation of keratinocytes, and from these observations it has been
proposed that vitamin C may be instrumental in the formation of the stratum corneum and may
thereby influence the ability of the skin to protect itself from water loss (Boyce et al., 2002).
Some studies have indicated that topical application of vitamin C may result in decreased
roughness, although this may depend more on the formulation of the cream than on the vitamin
C content. Because most studies in this area involve topical application, the complex and
variable effects (pH and additional compounds) of topical formulations make it difficult to come
to any firm conclusion as to whether vitamin C affects skin dryness.
 CHAPTER FIVE

CONCLUSION

5.1 Conclusion

The role of vitamin C in skin health has been under discussion since its discovery in the 1930s as
the remedy for scurvy. The co-factor role for collagen hydroxylases was the first vitamin C
function that was closely tied to the symptoms of scurvy and the realization of the importance of
this function for the maintenance of skin health throughout the human lifespan led to the
hypothesized skin health benefit of vitamin C. In addition, the antioxidant activity of vitamin C
made it an excellent candidate as a protective factor against UV irradiation. These two
hypotheses have driven most of the research into the role of vitamin C and skin health to date.
The following information is available as a result of research into the role of vitamin C in skin
health, and Tables 2 and 4 list a sample of key studies:

• Skin fibroblasts have an absolute dependence on vitamin C for the synthesis of collagen, and
for the regulation of the collagen/elastin balance in the dermis. There is ample in vitro data with
cultured cells demonstrating this dependency. In addition, vitamin C supplementation of animals
has shown improved collagen synthesis in vivo.

• Skin keratinocytes have the capacity to accumulate high concentrations of vitamin C, and this
in association with vitamin E affords protection against UV irradiation. This information is
available from in vitro studies with cultured cells, with supportive information from animal and
human studies.

• Analysis of keratinocytes in culture has shown that vitamin C influences gene expression of
antioxidant enzymes, the organization and accumulation of phospholipids, and promotes the
formation of the stratum corneum and the differentiation of the epithelium in general.

• Delivery of vitamin C into the skin via topical application remains challenging. Although some
human studies have suggested a beneficial effect with respect to UV irradiation protection, most
effective formulations contain both vitamins C and E, plus a delivery vehicle.
• Good skin health is positively associated with fruit and vegetable intake in a number of well-
executed intervention studies. The active component in the fruit and vegetables responsible for
the observed benefit is unidentified, and the effect is likely to be multi-factorial, although
vitamin C status is closely aligned with fruit and vegetable intake.

• Signs of aging in human skin can be ameliorated through the provision of vitamin C. A number
of studies support this, although measurement of skin changes is difficult. Some studies include
objective measures of collagen deposition and wrinkle depth.

• The provision of vitamin C to the skin greatly assists wound healing and minimises raised scar
formation. This has been demonstrated in numerous clinical studies in humans and animals.
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