TRADITIONAL USES OF PLANTS FOR CURING SKIN AILMENTS AND FOR COSMETIC PURPOSES

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SKIN STRUCTURE

The skin is the largest and most extensive organ of our body by surface area [1-3]. It accounts for about 15% of the total body by weight and covers the entire surface of the body [1, 4]. It serves as a physical barrier between the internal and the external environment [4, 5]. Further, it is part of the integumentary system maintaining an internal homeostatic balance [6]. It is our first line of defence against pathogens, irritants, trauma and ultraviolet radiation [2, 4, 7]. The skin synthesises vitamin D, provides temperature regulation and immunological protection, it is pivotal in the reception of physical stimuli from the outside environment [7]. Further, it enhances our body image and augments outward beauty which leads to a higher self-esteem [4, 8]. A healthy and beautiful skin is a major factor in the perception of general health and well-being of the body [2, 8, 9]. Consequently, it greatly contributes to the comfort and psychological well-being of an individual [6, 8]. The skin is made up of three major layers, which are the epidermis, dermis and hypodermis [1, 5, 10]. There is considerable regional variations in thickness of the human skin according to function and anatomic location [4, 6]. The skin ranges from about 0.5 mm in the eyelids and 3-4 mm in the soles of the feet with an average thickness of 1-2 mm in most parts of the body [1, 6].

Epidermis

The epidermis is the external skin surface, it is the thinner outer layer and is mainly made up of keratinocytes which constitute (90-95%), the remaining 5-10% is made up of Langerhans cells, Melanocytes and Merkel cells [1, 2, 4]. Melanin the pigment which protects the inner skin from UV radiation is produced in the epidermis by Melanocytes [6, 10]. Langerhans cells participate in immune response and Merkel cells function in the sensation of touch [6]. The epidermis is made up of five layers which are the stratum basale, stratum spinosum, stratum granulosum, stratum lucidum and the stratum corneum with each layer representing a stage in the life of an epidermal cell [6]. The structure and thickness of the epidermis varies according to anatomic location [3]. Its average thickness is 0.1 mm, but in acral areas (peripheral skin parts including palms, soles, elbows, knuckles, knees, toes, heels) it can go up to 1.6 mm thick [1, 3, 11]. The normal epidermis continuously undergoes renewal with keratinocytes on the skin surface being replaced after about 30 days [1, 4]. The epidermis is joined to the dermis through the dermal-epidermal junction which provides mechanical support for adhesion of the dermis to the epidermis [4, 5]. The dermal-epidermal junction is synthesised by dermal fibroblasts and basal keratinocytes [1, 4].

Dermis (corium)

The dermis is mainly made up of connective tissue and blood vessels, it is about 1 to 3 mm thick [5, 11]. Like the epidermis, the thickness of the dermis varies with anatomic location with the thickest layers being located in soles and palms and the thinner layers found in the eyelids [1, 11]. It has two layers the papillary (superficial dermis) and the reticular (deep) dermis [4, 6]. The papillary dermis contains nerves and capillaries used to nourish the epidermis, it provides adhesion between the dermis and the epidermis [1, 6]. The dermis is constituted of many cells (fibroblasts, dermal dendrocytes, mast cells), vessels and nerve endings [1, 4]. Fibroblasts are the most important cells found in the dermis and in all connective tissues because they synthesise all fibres and the ground substance of the dermis [1, 4]. The space between fibres and dermal cells is filled up by a ground substance which is made up of macromolecules which include proteoglycans (hyaluronic acid, dermatan sulphate, chondroitin-4-sulphate, fibronectin, tenascin, epimorphin) and glycoproteins [1, 4]. The ground substance is abundant in the papillary dermis [4]. The dermis contains two main types of sweat glands which are eccrine and apocrine glands, these release sweat into hair follicles on the skin surface through pores [1, 6]. Further, the superficial dermis is constituted of collagen fibres in loose bundles and thin elastic fibres which stretch to the dermal-epidermal junction [1, 4]. In contrast, the reticular dermis has coarser collagen bundles arranged parallel to the skin surface and a thicker elastic network, additionally it contains the deep part of cutaneous appendages, vascular and nerve plexuses [1, 4].

Collagen and elastin fibres

Collagen fibers which are produced from fibroblasts in the skin dermis are the major constituent of the skin dermis constituting more than 90% wet weight and 98% of total mass of dried dermis [1, 10, 12]. The fibres are made up of mainly type I and type III collagen [4]. Collagen fibres are arranged in loose bundles in the papillary dermis and become thicker in the deep dermis [1]. The protein is the chief structural unit responsible for tensile strength, mechanical resistance and rigidity of the skin [1, 2, 13, 14]. It ensures cohesion and regeneration of the skin, cartilage and bone [15]. It is important in controlling cell shape and differentiation, migration, synthesis of some proteins and acts as a base on which other cells can proliferate [15]. Elastin fibres like collagen fibres are produced in the skin dermis by connective tissue cells known as fibroblasts [10]. Elastin is responsible for elasticity and retractile properties, in addition it provides resilience of the skin [8]. The protein is most abundant in organs providing elasticity to the connective tissues [10]. Elastin fibres are thin in the papillary dermis, they become thicker and are arranged horizontally in the reticular dermis [4].

The hypodermis (subcutaneous fat, panniculus adiposus)

The hypodermis is a connective tissue binding the skin to internal organs and is the thickest layer of the skin [3, 5]. It is an adipose tissue which is important in regulating temperature, insulation, protection from mechanical injuries and it acts as a store of energy [1, 3]. Adipocytes are the major cells in this tissue, they are large, rounded cells whose cytoplasm is composed of triglycerides and fatty acids [1]. These cells are arranged in lobules separated by connective tissue septae, the thickness of the hypodermis shows anatomic variation between individuals and is reflective of nutritional status of an individual [3].

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Table of contents :

Submission declaration
Plagiarism declaration
Acknowledgements
Summary
List of Figures
List of Tables
Abbreviations
Supplementary data
CHAPTER 1: INTRODUCTION
- 1.1 SKIN STRUCTURE
- 1.1.1 Epidermis
- 1.1.2 Dermis (corium)
- 1.1.2.1 Collagen and elastin fibres
- 1.1.3 The hypodermis (subcutaneous fat, panniculus adiposus)
- 1.2 SKIN AGING
- 1.2.1 Role of elastase and collagenase in skin aging
- 1.2.2 Existing anti-aging products and their side effects
- 1.2.2.1 Sunscreens
- 1.2.2.2 Hydroquinones
- 1.2.2.4 Hormones
- 1.2.2.5 Alpha hydroxy acids and alpha keto acids
- 1.3 NATURAL PRODUCTS AS ANTI-AGING INGREDIENTS
- 1.3.1 Natural antioxidants
- 1.3.2 Retinoids
- 1.3.3 Vitamin D
- 1.3.4 Vitamin C
- 1.4 TRADITIONAL USES OF PLANTS FOR CURING SKIN AILMENTS AND FOR COSMETIC PURPOSES
- 1.5 PHYTOCHEMICALS AS INHIBITORS OF COLLAGENASE AND ELASTASE
- 1.6 PHYTOCHEMICALS AS POTENTIAL GROWTH PROMOTERS OF ELASTIN AND COLLAGEN
- 1.7 PROBLEM STATEMENT AND JUSTIFICATION
- 1.8 GOALS AND OBJECTIVES
- 1.9 REFERENCES
CHAPTER 2: Sclerocarya birrea (MARULA)
- 2.1 BOTANY AND SPATIAL DISTRIBUTION
- 2.2 TRADITIONAL USES OF THE PLANT
- 2.3 COSMETIC APPLICATIONS
- 2.4 PREVIOUSLY REPORTED BIOASSAYING OF MARULA
- 2.5 PHYTOCHEMISTRY
- 2.5.1 Marula stems
- 2.5.2 Marula fruit and seed
- 2.5.3 Marula leaves
- 2.6 METHODOLOGY
- 2.6.1 Collection and extraction of Marula stems, leaves and fruits
- 2.6.2 Collection and extraction of Marula stems
- 2.6.3 Sequential extraction of Marula stems
- 2.6.4 Marula oil production
- 2.6.5 Bioassay of crude extracts and fractions
- 2.6.6 Chemical Profiling
- 2.6.7 Bioassay-guided fractionation
- 2.6.8 Defatting of Marula stem ethanol extract
- 2.6.9 Concentration of actives of Marula stem extract
- 2.6.10 Defatting-concentration of actives of Marula stem extract
- 2.7 RESULTS AND DISCUSSION
- 2.7.1 Selection of the most appropriate solvent
- 2.7.2 Sequential extraction using polar and non-polar solvents and bioassaying
- 2.7.3 UPLC-QTOF-MS analysis of Marula stems extracted sequentially using polar
- and non-polar solvents
- 2.7.4 Separate extraction using different polar solvents and their bioassaying
- 2.7.5 UPLC-QTOF-MS analysis of Marula stems extracted separately with different solvents
- 2.7.6 Chemical profiling and phytochemical analysis of the ethanol extract of Marula stems
- 2.7.7 Improving the quality of the ethanol extract as a cosmetic ingredient
- 2.7.8 Comparison of chemical profiles of the Marula extract, and fractions obtained through defatting and concentration
- 2.7.9 Bioassay-guided fractionation
- 2.7.10 Collagenase inhibition activity of fractions of S. birrea
- 2.7.11 UPLC-QTOF-MS analysis of active fractions
- 2.7.12 Collagenase inhibition assaying of pure compounds
- 2.7.13 Elastase inhibition activity of fractions of S. birrea
- 2.7.13.1 UPLC-QTOF-MS analysis of active fractions
- 2.7.14 Elastase inhibition of pure compounds
- 2.8 CONCLUSIONS
- 2.9 REFERENCES
Chapter 3: Ficus sycomorus (SYCAMORE)
- 3.1 BOTANY AND GEOGRAPHICAL DISTRIBUTION
- 3.2 TRADITIONAL USES OF THE PLANT
- 3.3 PREVIOUS RESEARCH ON F. SYCOMORUS
- 3.3.1 Leaves
- 3.3.2 Stem bark
- 3.3.3 Roots
- 3.3.4 Fruits
- 3.5 PHYTOCHEMISTRY
- 3.5.1 Stems
- 3.5.2 Leaves
- 3.5.3 Roots
- 3.6 METHODOLOGY
- 3.6.1 Collection and extraction of plant material from KwaZulu Natal
- 3.6.2 Collection of plant material from the University of Pretoria garden and experimental farm
- 3.6.3 Separate extraction of leaf material using polar solvents
- 3.6.4 Sequential extraction of leaf material
- 3.6.5 Bioassaying of crude extracts and fractions
- 3.6.6 Chemical profiling using UPLC MS and NMR
- 3.6.7 Bioassay guided fractionation
- 3.6.8 Defatting the crude ethanol extract of leaves of F. sycomorus
- 3.7 RESULTS AND DISCUSSION
- 3.7.1 Sequential extraction using polar and non-polar solvents and bio-assaying
- 3.7.2 UPLC-QTOF-MS analysis of F. sycomorus leaves extracted sequentially with polar and non-polar solvents
- 3.7.3 Separate extraction using different polar solvents and their bioassaying
- 3.7.4 UPLC-QTOF-MS analysis of F. sycomorus leaves extracted separately with polar solvents
- 3.7.5 Chemical marker identification and phytochemical analysis
- 3.7.6 Fractionation of ethanol extract of leaves of F. sycomorus
- 3.7.7 Structure elucidation using NMR
- 3.7.8 Improving the quality of the ethanol extract as a cosmetic ingredient
- 3.7.9 Comparison of chemical profiles of the F. sycomorus crude extract and its defatted fraction
- 3.7.10 Bioassay guided fractionation
- 3.7.11 Collagenase inhibition activity of fractions of F. sycomorus
- 3.7.12 UPLC-QTOF-MS analysis of active fractions
- 3.7.13 Collagenase inhibition assay of pure compounds
- 3.7.14 Elastase inhibition activity of fractions of F. sycomorus
- 3.7.15 UPLC-QTOF-MS analysis of active fractions
- 3.7.16 Elastase inhibition activity of pure compounds
- 3.8 CONCLUSIONS
- 3.9 REFERENCES
CHAPTER 4: COMBINATION AND FORMULATION STUDIES
- 4.1 INTRODUCTION
- 4.2 COMBINATION STUDIES
- 4.3 FORMULATION STUDIES
- 4.4 METHODOLOGY
- 4.4.1 Preparation of mixtures
- 4.4.2 Bioassay of the combinations/mixtures
- 4.4.3 Formulation of the anti-aging day cream at lab scale
- 4.4.4 Formulation of the nourishing night cream at lab scale
- 4.4.5 Formulation of the anti-aging day cream at kilogram scale
- 4.4.6 Formulation of nourishing night cream at kilogram scale
- 4.5 RESULTS AND DISCUSSION
- 4.5.1 Bioassay of the combinations/mixtures
- 4.5.1.1 Collagenase inhibition activity
- 4.5.1.2 Elastase inhibition activity
- 4.5.2 Anti-aging day cream-SPF
- 4.5.3 Nourishing night cream
- 4.6 CONCLUSION
- 4.7 REFERENCES
CHAPTER 5: METHODOLOGY
- 5.1 INTRODUCTION
- 5.2 CHAPTERS 2 AND
- 5.2.1 Chemicals and reagents
- 5.2.2 Extraction procedures
- 5.2.2.1 Extraction of plant material at CSIR
- 5.2.2.2 Separate extraction of plant material
- 5.2.2.3 Sequential extraction of plant material
- 5.2.3 Defatting of extracts
- 5.2.4 Concentration of actives
- 5.2.5 Defatting-concentration of actives
- 5.2.6 Determination of anti-elastase activity
- 5.2.7 Determination of anti-collagenase activity
- 5.2.8 Statistical Analysis
- 5.2.9 UPLC-Q-TOF-MS analysis
- 5.2.9.1 MS Conditions
- 5.2.9.2 Acquisition
- 5.2.10 Column and thin layer chromatography
- 5.2.11 Purification of compounds 1, 2 and 3 using preparative HPLC-MS
- 5.2.12 NMR
- 5.2.13 Chemical profiling
- 5.2.14 Semi Prep HPLC fractionation
- 5.3 CHAPTER
- 5.3.1 Combination studies
- 5.3.1.1 Combination of Marula and Sycamore extracts
- 5.3.1.2 Combination of Sycamore extract and active compounds from Marula
- 5.3.2 Formulation studies
- 5.3.2.1 Formulation of the antiaging day cream (200.00 g batch)
- 5.3.2 Formulation of the nourishing night cream (200.00 g batch)
- 5.3.3 Formulation of the antiaging day cream (2 kg batch)
- 5.3.4 Formulation of the nourishing night cream (2 kg batch)
- 5.4 REFERENCES
CHAPTER 6: CONCLUSIONS