(NOVA) as a Substitute for Routine Fixatives in Cervical Smear. J

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1 JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL SCIENCES Idoko J, Enemari JO, Okolo SC, Yaro DJ, Akpulu P, Mairiga AA et al.. Hairspray (NOVA) as a Substitute for Routine Fixatives in Cervical Smear. J Pharm Biomed Sci 214; 4(4): The online version of this article, along with updated information and services, is located on the World Wide Web at: Journal of Pharmaceutical and Biomedical Sciences (J Pharm Biomed Sci.), Member journal. Committee of Publication ethics (COPE) and Journal donation project (JDP).

2 ISSN NO Research article Hairspray (NOVA) as a Substitute for Routine Fixatives in Cervical Smear Idoko J 1,*, Enemari JO 1, Okolo SC 1, Yaro DJ 1, Akpulu, P 2, Mairiga AA 1, Aghenumu L 1, Owolagba G 1, Dasi KR 3,Anyebe EE 4, Ameh EA 5, Ogboi SJ 3 Affiliation:- 1 Department of Pathology Ahmadu Bello University Teaching Hospital Zaria 2 Department of Human Anatomy Ahmadu Bello University Zaria 3 Research and Training Unit, Department of Nurse Education, ABU Teaching Hospital Zaria. 4 Paediatric Surgery Unit, Department of Surgery, A.B.U. Teaching Hospital Zaria, Nigeria 5 Public Health Lab. Department of Med. comm. ABU Zaria The name of the department(s) and institution(s) to which the work should be attributed: 1.Department of Pathology Ahmadu Bello University Teaching Hospital Zaria. 2.Department of Human Anatomy Ahmadu Bello University Zaria. 3.Research and Training Unit, Department of Nurse Education, ABU Teaching Hospital Zaria. 4.Paediatric Surgery Unit, Department of Surgery, A.B.U. Teaching Hospital Zaria, Nigeria 5.Public Health Lab. Dept of Med. comm. ABU Zaria. *To whom it corresponds:- Idoko J. Department of Pathology Ahmadu Bello University Teaching Hospital Zaria. Contact number: jonidos21@yahoo.com Abstract: High incidence of death has been associated with cervical cancer in the developing countries in the recent times, and so it is imperative to advance its detection. Hairspray (NOVA) was compared with routine 95% ethanol to assess the efficiency in cyto diagnosis of cervical smear. Two hundred (2) cervical smeared samples in duplicate were collected from patients, carefully labeled and were fixed immediately, using routine 95% ethanol and with hairspray (Nova) respectively. The slides were stained by papanicolaou technique and were evaluated. There was no degeneration of cell morphology and no significant toxic effect of the Hairspray (NOVA) when compared with 95% alcohol. The cells appeared normal, without shrinkage and with a clear background and good nuclear outline. Hairspray (NOVA) is cost effective, efficient, handy and easy to use. It is shown that, hairspray (Nova) is effective for cervical smear. Keywords: Hairspray; routine fixative; cervical smear. Article citation: Idoko J, Enemari JO, Okolo SC, Yaro DJ, Akpulu P, Mairiga AA et al. Hairspray (NOVA) as a substitute for Routine fixatives in cervical smear. J Pharm Biomed Sci 214; 4(4): Available at INTRODUCTION Cervical cytological screening among women has become an acceptable phenomenon of detecting precancerous and cancerous disease and this have gone a long way to reduce death resulting from cervical cancer 7,5. The costs of such cytological screening services are high in most developing nation 8. As a standard, most laboratory adopt fixation of smears immediately in 95% alcohol, for 15minutes to 1 hour.95% ethyl alcohol is satisfactorily, but addition of 3% glacial ascetic acid increase the nucleo-protein fixing properties and gives excellent nuclear and cytoplasmic morphology 2. Fixatives of different types including aerosol 327

3 ISSN NO sprays could be used to preserve the cells structure and architecture. Most aerosol fixatives as in the case of Hairspray (NOVA) is alcohol base containing mixtures of alcohols, acetic acid and propylene or ethylene glycols for the preservation and transportation of smear, and increases the diagnostic utility of cervical smear 1. These spray fixatives provides a waxy coat on the slide, thus maintaining cellular integrity and the architecture 6.Transportation and mailing fixed slides using spray aerosol is easier and convenient. Nevertheless the routine fixative is mostly used in most of the laboratory because of its availability, though Hairspray a type of aerosol has been noted to be effective and cheap cytological fixative 11. In correct fixations are common due to inappropriate training of workers, or heavy workload. That leads to repeating smears, and increasing the workload or missing the patient s results. One alternative method to overcome this problem is to introduce hairspray as a fixative. Therefore, the aim of this study is to compare fixative properties of hairspray (NOVA) with 95% alcohol and to determine the cost effectiveness of hairspray. MATERIALS AND METHODS STUDY POPULATION Two hundred (2) smeared samples were collected from patients in duplicate, they were labeled and immediately fixed as A and B, where A is fixation in hairspray NOVA and B is fixationin routine 95% ethanol. The slides were stained by papanicolaou technique and were screened by nine Medical Laboratory Scientists using X 2 and x 4Leica microscope as shown in table 1. The Medical Laboratory scientists completed a form stating whether fixation was good or bad, considering the intactness of the cells organelle, attachment to the slide, folding, over or under stain, cloudiness. METHODS OF COLLECTION Sexually active women samples were used, vaginal mucosa examined and speculum inserted to dilate and expose the cervix. Ayre s spatula was used to scrape the cervical mucosa and entire surface of the external and internal OS. The scrapping was then smeared on the two labeled slides. The slides were immediately sprayed with Hairspray Nova and the other dropped into the jar containing 95% ethanol. Fixed smeared were left in the fixative for at least 15minutes before staining to avoid drying. STAINING Papanicolaou staining method is employed after fixation through descending grades of ethanol as cited by Bancroft 21. Stained in Harris Haematoxylin for 5 minutes and afterwards rinse in tap running water, then blue in ammoniacal water. Rinse in tap running water and transfer to different changes of alcohol, stain in OG6 for two minutes. Rinse in changes of alcohol and stain in EA36 for 2minutes. Rinse in 95% alcohol then absolute alcohol and clear in xylene and mount using DPX. RESULTS The cellular structure observed for hairspray (Nova) and the routine 95% alcohol. CELL MORPHOLOGY There is no cytoplasmic shrinkage, all the cells were preserved, the nuclear are intact, the nuclear detail is clear and good as evidence in Fig 1, 2 and table of result. SUPERFICIAL CELLS The slides viewed were all normal. Well outlined nuclear appearance, no cytoplasmic shrinkage as shown in fig.1, 2 and table 1. INTERMEDIATE CELLS slides viewed were normal, with no shrinkages, well outlined nuclear appearance and % clarity as shown in fig.1, 2 and table 1. PARABASAL CELLS slides viewed were all normal, no shrinkage and % nuclear clarity as shown on fig.1, 2 and table1. STAINABILITY It turns out good as exemplified in pap staining method as shown in fig.1 and

4 ISSN NO Table 1. Comparison between routine fixatives and hairspray. Status Routine fixatives (No of Slides = ) Hairspray (No of Slides = ) Superficial Cells Inter-mediate Parabasal Cells Superficial Cells Inter-mediate Parabasal Cells Good (+) Bad (-) Figure 1. Photomicrograph of cervical smear fixed with Hairspray. Note the well outlined nuclear appearance (N),Papanicolaou Stain MgX 2.Leica microscope 4. Figure 2. Photomicrograph of Hairspray fixed cervical smear, showing normal morphology(superficial S, intermediate I, parabasal P) and having a normal background. Papanicolaou Stain MgX 2.Leica microscope 4. Figure 3. Photomicrograph of a section of showing same sample of cervical smear as in figure 1 and 2 but fixed in 95% ethyl alcohol. Note the preserved RBC s (R) obscuring epithelial cells. (EC).Papanicolaou Stain, MgX 2.Leica microscope 4. DISCUSSION The use of commonly available women s Hairspray such as Fencot spray as a fixative for cytodiagnosis is reported in the year 1969 by freeman. Hairspray has been confirmed to be as effective as fixative for cytodiagnosis as a commercially available aerosol fixative 11. It also gives a prerequisite for convenience of transport or mailing of slides. Though aerosol causes shrinkage of cell morphology which was noted by Utian Wulf H. 11 but in this work no significant shrinkage was noted in the nucleus or cytoplasm. The hairspray was extremely effective in maintaining the morphology of cells as seen in fig.2. Hairspray obviated the necessity of any repeat smear, commonly resulting from cells being washed off the slide during staining after 95% ethanol fixation. The use of hairspray will eliminate the issue of contamination due to fixing in the same jar and reusing of alcohol. From the result above, hairspray (Nova) which constitutes 329

5 ISSN NO includes butane, lsobutane, propane, Aqua, VA/crotonates/vinyl codecanoatecopolymer, Aminomethylpropanol, PEG-12, Dimethicone, parfum,benzophenone-3, panthenol,linaloel is as effective as routine fixative and this tally with the work done by Freeman in It fixed the superficial, intermediate and parabasal cells excellently, with good nuclear-cytoplasmic staining. It does not constitute any ingredients that will destroy red blood cells. Much more it increases the visualization of diagnostic cellular material. Thus, it will bring to the barest minimum the incidence and mortality of invasive disease. The nuclear and cytoplasm staining of smear production is clear and in good contrast as attained in furtherance of other advance histological procedures as stated by Innovatek 23. Hairspray (Nova) gives clear background well outlined nuclear appearance as seen in fig. 1. Fig.3 is the smear fixed with (95% alcohol) routine fixative. In fig. 1, 2 & 3 shows that further subsequent treatments after fixation did not affected the cell morphology and this clearly shows that it has competitive property of a good fixative. The cost of Ethanol (Routine 95%) is two thousand five naira per 625 liters and sufficiently serves 31 samples that are 2mls to each specimen container. The cost of the Hair spray fixatives (NOVA) is three hundred and fifty naira per 35ml and sufficiently use for 2 cervical samples. This is relatively cheap as compared with the routine 95% alcohol as per the use of one slide in single chamber and much more causing floaters when multiple slides in one chamber. Floater which is the transfer of one particle from sample to other has been a menace in histopathological diagnosis. Aerosol fixative is characterized to cause shrinkage of the cell morphology and particularly of the nucleus, in other words it results in inadequate and in accuracy of Hormonal evaluation 1. It has been noted that cytology fixative spray preserved smears for the detection of HPV, HIV, genomic mutations and other viral and bacterial pathogen. By extension, it also stabilizes good antigenic preservation for routine staining and other advanced test like immunohistochemistry, FISH (Innovatek Medical Inc.,) 4. The hairspray Aerosol spray fixatives (NOVA) used for this work favours cervical and vaginal pool smear it can be advocated to be use in non gynaecological smear or buccal smears for nuclear chromatic assessment, urine, pericardial effusion, cerebrospinal fluid, joint fluid and peritoneal fluid. CONCLUSION Hairspray (Nova) is an effective, efficient, handy and easy to use fixative for cervical smear. It is very good when transporting smear samples. RECOMMENDATIONS The hairspray Aerosol spray fixative (NOVA) is recommended to be incorporated into routine cytopathological procedure. REFERENCES 1. Baharom Anisah, Ismail Maimunah. Case Studies on Decision for Cervical Cancer Screening among Working Women The Journal of Human Resource and Adult Learning,28:4(2): Bancroft J D and Stevens (21) Theory and Practice of Histopathological Techniques 3 rd ed P 466. Churchhill Livingstone London, New York. 3. Freeman, J. A. (1969): Fixation of smears using cytofixative Acta Cytol.(philad.),13, Innovatek Medical Inc. ( 23) #3-16 Derwent way, Delta Canada. V3MS6M5. 5. Kumar, Vinar; Abbas, Abul k, Fausto, Nelson; Mitchell, Richard N. (27). Robbins Basic Pathology (8 th ed.) sauders Elsevier. London. 6. Kumarasinghe M P, Constantine T S R, Hemarnali T R LC. Methanol as an alternative fixative for cytological smears, Malaysian J Patlzol,1997;19(2): Lim G.C.C. Halimah Y. (24) Second Report of the National Cancer Registry. Cancer Incidence in Malaysia 23. National Cancer Registrt. Kuala Lumpur. 8. Markovic, M., Kesic V., Topic L., Matejic, B. Barriers to cervical cancer screening: a qualitative study with women in Serbia. Social science and medicine,25; 61: Ramzi SC Stanley RL, Vinary K (1999) Disease of The Female Genital Tract in pathologic basis of disease 6 th ed. Published by Saunders philadephia London Toronto Mexico city. 1. Shamsi M, Abdali K, Montazer NR, Kumar PV, Tabatabaee HR. Comparison of Carnoy's solution and 96% ethyl alcohol fixation in bloody Pap smears.acta Cytol. 23;47(4): Utian, Wulf H. Hairspray as an Inexpensive Fixative for Papanicolaou Smears South Africa medical journal, 1972;

6 ISSN NO Source of support: None Competing interest / Conflict of interest The author(s) have no competing interests for financial support, publication of this research, patents and royalties through this collaborative research. All authors were equally involved in discussed research work. There is no financial conflict with the subject matter discussed in the manuscript. Copyright 214 Idoko J, Enemari JO, Okolo SC, Yaro DJ, Akpulu P, Mairiga AA et al.. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 331