ABSTRACT
Two plant extracts: aqs. Lawsoniainermis and aqs.Cnestisferruginea were investigated as substitute dyes against hematoxylin and eosin staining technique. Two methods were employed which are aqs. Lawsoniainermis& eosin staining technique and hematoxylin&aqs.Cnestisferruginea staining technique, they were used firstly to determine their staining potentials. Two groups of liver and kidney tissue sections were used in this investigation , hence section showed distinctive features after being stained with direct comparison with being stained with the widely known H&E staining technique. This shows that aqs. Lawsoniainermis can be used as a suitable substitute for hematoxylin stain while aqs. Cnestisferruginea can also be used as a replacement for the eosin stain.
CHAPTER ONE
1.0 INTRODUCTION
The human body is made up of various tissues with different functions. Structure is closely related to function and much can be deduced about the function of cells and tissues by careful examination of their component part (Young et al ., 2014). In case of abnormality, the tissues can be studied and prompt diagnosis carried out. Stains help differentiate the various structures present in both normal and abnormal tissues thus, accurate diagnosis can be achievable. Abnormal tissues are known as pathological tissues and can be significantly differentiated from the normal tissues by the use of stains Staining is a process by which we give color to a section.Generally the stains are classified as:Acid stains, Basic stainsandNeutral stains (Talukder, 2008).Staining techniques originated from the second half of the last century. Stains have been used to enhance accurate descriptions of the microscopic structure of tissues, which is necessary for histopathologic diagnosis. Plant and insect parts have found place in histological staining due to their colouring and dying effect. Examples of plant and insect parts that have found place in histological staining as natural dyes are Haematoxyloncampechiaumn, from which haematoxylin is obtained and Dactylopiuscacti, from which carmine stain is obtained. Although most of the dyes in current use in histopathology laboratories are of synthetic origin, natural dyes still hold promise as a potential source cheaper dyes and consequently providing employment opportunities in developing countries. (Egbujoet al ., 2008).
Several reports declared that lawsone is the major constituent of Lawsoniainermis and can treat tuberculosis. From Pakistan such information has been compiled and reported showing that lawsone is useful against oral Candida albicansseperated from patients with HIV/AIDS among human. Growth and replication of retroviruses (HIV) were found to be inhibited through naphthoquinones. demonstrated anticancer trait of Henna’s using an extract of Lawsoniainermis in chloroform by the culture tetrazollium salt (MTT) test on the breast of human, liver and colon normal cell lines and carcinogenic liver cell lines .Lawsoniainermis(Lythraceae) is a perennial plant commonly called as henna,L. inermis is widely distributed throughout the Sahel and into Central Africa. It grows mainly along watercourses and in semi-arid regions and is adapted to a wide range of conditions. It can withstand low air humidity and drought. Henna requires high temperatures for germination, growth and development. Henna leaves are very popular natural dye to color hand, finger, nails and hair. The dye molecule, Lawson is the chief constituents of the plant; its highest concentration is detected in the petioles (0.5-1.5%). In folk medicines, henna has been used as astringent, antihemorrhagic, intestinal antineoplastic, cardio-inhibitory, hypotensive, sedative and also as therapeutic against amoebiasis, headache, jaundice and leprosy (Kannahi and Vinotha, 2013).Lawsoniainermis,is a medicinal plant. According to a research conducted by Azubike et al. (2010), the staining ability of the lime juice extract [LJE] and Methanol extract [ME] of Lawsoniainermis leaves on animal histological sections was demonstrated. When compared with the control. L. inermis extracts [LJE and ME] could stain tissues effectively and may be used in the future to replace eosin in H & E staining technique (Azubikeet al.,2010).
Cnestisferrugineaon the other hand is widely used as a medicine in Africa. There has been little research into its constituents, but the leaves, bark and roots have been recorded as containing tannins, whilst the presence of glycoside may also be suspected, leaf-sap is placed on the eyelids and instilled into the eyes in the treatment of eye-troubles The leaves, or the roots, are used for treating dysmenorrhoea A paste of the root-bark is rubbed on the forehead for treating headaches. The roots are used as a purgative in Nigeria and some african countries. The roots enter into remedies for treating skin-infections, often applied as an ointment, from studies by Akpo (2013), has shown that Cnestisferrugineahas action against Sarcinalutea and Staphylococcus aureus, but no action against Gram -ve organisms, nor fungi. The fruit pulp is taken as a tonic, and is used to treat bronchial affections, especially whooping-cough and tuberculosis. The fruit pulp is rubbed on the skin and is used as a medicine for the throat. The juice is used as an eye-instillation for various eye-complaints, principally conjunctivitis. The juice is applied to wounds. The fruit, together with the seeds, is ground up with alcohol or boiled in wine to produce a remedy for snake-biteThe bitter fruits are used to clean the teeth The fruit contains a soft, juicy, somewhat bitter and acid pulp. This is widely used in many parts of Africa to rub on the teeth to clean and whiten them. It leaves a refreshing taste in the mouth. The bark yields a red dye which is used for dyeing clothing (Fern, 2014).Staining of pathological samples can be carried out using these natural stains since most of the stains used on pathological samples are costly, and not readily available. Some are toxic and not environmental friendly (James, 2005).
1.2 JUSTIFICATION OF STUDY
Most synthetic stains are costly and toxic (James, 2005). This research tries to compare the routine haematoxylin and eosin stain and extracts from Lawsoniainermisandcnestisferrugineaasstains for pathological tissues, Evaluating their cost effectiveness and non-toxicity.
1.3 AIM AND OBJECTIVES
- To determine the effect of Lawsoniainermis and cnestisferrugineaon pathological tissues
- To compare and evaluate routine haematoxylin and eosin stain and Lawsoniainermis and
- To investigate the possibilities of replacement of routine stains with Lawsoniainermisand
1.4 RESEARCH OBJECTIVES
The main objectives of this study is to
* Determine if the extracts have staining potentials
* Know the color pickups of components of the organs stained
* Determine if the extracts are suitable substitute for hematoxylin and eosin stain
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