Measurement of Pterygium Tissue Dry Weight Using Two Different Tissue Preparation Techniques in Freeze-Dry Method

Mohd Radzi Hilmi  -  Department of Optometry and Visual Sciences, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia (IIUM)
Khairidzan Mohd Kamal*  -  Department of Ophthalmology, Kulliyyah of Medicine International Islamic University Malaysia (IIUM)
Mohd Zulfaezal Che Azemin  -  Department of Optometry and Visual Sciences, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia (IIUM)
Azrin Esmady Ariffin  -  Faculty of Optometry and Vision Science, SEGi University, Petaling Jaya, Selangor, Malaysia

(*) Corresponding Author

Due to pterygium tissue compression on the corneal surface, it has been hypothesised that the degree of corneal astigmatism caused by pterygium can vary. This study aims to develop a freeze-dried method for estimating net pterygium tissue mass (NPTM) as dry weight. A single surgeon (KMK) excised 60 primary pterygium using the controlled partial avulsion technique and divided them into two groups: formalin-fixed (n=30) and unfixed (n=30). After determining the weight of each sterile container, 5 mL of 5% buffered formaldehyde was added to the formalin-fixed group and stored for one week, while 5 mL of distilled water was added to the non-fixed pterygium group. Each container was pre-frozen for 12 hours before being freeze-dried (-24 hours at -80 degrees Celsius). The result is referred to as the net pterygium tissue mass (NPTM). Using an independent T-test, a comparison of wet and dry weight and percentage of NPTM was conducted between groups. Wet weights for formalin-fixed and non-fixed pterygium were 253.33 82.17 g and 255.17 63.52 g, respectively, while dry weights were 184.92 84.31 g and 179.54 72.85 g. Formalin-fixed pterygium tissue revealed a slightly higher percentage of NPTM than non-fixed pterygium tissue (69.39 13.29% vs. 67.75 13.29%, p = 0.792), but this difference was not statistically significant. The freeze-dried method can be utilised to quantify the NPTM of pterygium fibrovascular tissue and investigate the influence of pterygium translucency on predicting induced-corneal astigmatism.

Keywords: Pterygium; Freeze-dried; Dry weight; Corneal astigmatism

  1. Allen, C. L., Clare, G., Stewart, E. A., Branch, M. J., McIntosh, O. D., Dadhwal, M., Dua, H. S., & Hopkinson, A. (2013). Augmented Dried versus Cryopreserved Amniotic Membrane as an Ocular Surface Dressing. PLOS ONE, 8(10), e78441.
  2. Ang, Leonard PK., Chua, Jocelyn LL., Tan, Donald TH. (2007). Current concepts and techniques in pterygium treatment. Current Opinion in Ophthalmology 18(4): 308-313. DOI: 10.1097/ICU.0b013e3281a7ecbb
  3. Azemin, M.Z.C, Hilmi, M.R., Kamal, K.M. (2014). Supervised Pterygium Fibrovascular Redness Grading Using Generalized Regression Neural Network,
  4. Frontiers in Artificial Intelligence and Applications, 265: 650-656. DOI10.3233/978-1-61499-434-3-65
  5. Azemin, M.Z.C. et al., 2015. GLCM texture analysis on different color space for pterygium grading, ARPN Journal of Engineering and Applied Sciences, 10: 6410-6413.
  6. Azemin, MZC., Tamrin, Izzuddin M., Hilmi, MR., Kamal, KM. (2016). Inter-grader reliability of a supervised pterygium redness grading system, Advanced Science Letters, 22(10): 2885-2888.
  7. Benders, K.E., Malda, J., Saris, D.B.F., Dhert, W.J.A., Steck, R., Hutmacher, D.W., Klein, T.J. (2010). Formalin fixation affects equilibrium partitioning of an ionic contrast agent-microcomputed tomography (EPIC-μCT) imaging of osteochondral samples, Osteoarthritis Cartilage 18:1586-1591.
  8. Birkl, C., Langkammer, C., Golob-Schwarzl, N., Leoni, M., Haybaeck, J., Goessler, W., Fazekas, F., and Ropele, S. (2016). Effects of formalin fixation and temperature on MR relaxation times in the human brain. NMR in Biomedicine, 29: 458– 465. doi: 10.1002/nbm.3477.
  9. Chen, T., Ding, L., Shan, G., Ke, L., Ma, J., Zhong, Y. (2015) Prevalence and Racial Differences in Pterygium: A Cross-Sectional Study in Han and Uygur Adults in Xinjiang, China. Investigative Ophthalmology & Visual Science, 56(2):1109-1117. doi:
  10. Connon CJ., Doutch J., Chen B., Hopkinson, A., Nakamura, T., Kinoshita, S., Meek, KM. (2010). The variation in transparency of amniotic membrane used in ocular surface regeneration. British Journal of Ophthalmology 94:1057-1061.
  11. Džunić, B., Jovanović, P., Veselinović, D., Petrović, A., Stefanović, I., & KovaÄević, I. (2010). Analysis of pathohistological characteristics of pterygium. Bosnian Journal of Basic Medical Sciences 10(4): 307-303.
  12. Errais K, Bouden J, Mili-Boussen I, Anane R, Beltaif O, Meddeb Ouertani A. (2008). Effect of Pterygium Surgery on Corneal Topography. European Journal of Ophthalmology. 18(2):177-181. doi:10.1177/112067210801800203
  13. Frota, A. C. de A., Lima Filho, A. A. de S., Dias, A. B. T., Lourenço, A. C. dos S., Antecka, E., & Burnier Jr., M. N.. (2008). Freeze-drying as an alternative method of human sclera preservation. Arquivos Brasileiros De Oftalmologia, 71(2): 137–141.
  14. George, D. & Mallery, M., 2010, SPSS for Windows Step by Step, A Simple Guide and References, Boston: Pearson.
  15. Hilmi MR., Che Azemin MZ., Mohd Kamal K., Mohd Tamrin MI., Abdul Gaffur N., Tengku Sembok TM. (2017). Prediction of Changes in Visual Acuity and Contrast Sensitivity Function by Tissue Redness after Pterygium Surgery. Current Eye Research. 2017 42(6):852-856. doi: 10.1080/02713683.2016.1250277.
  16. Hilmi MR., Kamal, KM., Azemin, MZC., Ithin, MH., Mystafa, MMSM., Yusof, F., Ahmad, N. (2018). Corneal Curvature Measurements Utilizing a New Swept-source Optical Coherence Tomography Tomey OA-2000® and Comparison With IOL Master® 500 in Pterygium Patients. Sains Medika: Jurnal Kedokteran dan Kesehatan, 9(1):11-17. doi:
  17. Hilmi MR., Khairidzan MK., Azemin ZC., Azami MH., Ariffin, AE. (2018). Measurement of Contrast Sensitivity Using the M&S Smart System II Compared with the Standard Pelli–Robson Chart in Patients with Primary Pterygium. Makara Journal of Health Research 22(3): 169-171.
  18. Hilmi, M.R., Khairidzan, M. K., Mohd Zulfaezal, C. A., & Azrin, E. A. (2019). Corneo-pterygium total area measurements utilising image analysis method. Journal of Optometry 12(4): 272-277.
  19. Jiao, W., Chengchao Zhou, Ting Wang, Shaoyuan Yang, Hongsheng Bi, Liping Liu, Yan Li, Lihua Wang. (2014). Prevalence and Risk Factors for Pterygium in Rural Older Adults in Shandong Province of China: A Cross-Sectional Study. BioMed Research International, Vol. 2014, Article ID 658648: 1-8.
  20. Kumagai, A., Kondo, F., Sano, K., Inoue, M., Fujii, T., Hashimoto, M., Watanabe, M., Soejima, Y., Ishida, T., Tokairin, T., Saito, K., Sasajima, Y., Takahashi, Y., Uozaki, H., & Fukusato, T. (2016). Immunohistochemical study of hepatocyte, cholangiocyte and stem cell markers of hepatocellular carcinoma: the second report: relationship with tumor size and cell differentiation. Journal of Hepato-Biliary-Pancreatic Sciences, 23(7), 414-421.
  21. Libera, R. D., Melo, G. B. de ., Lima, A. de S., Haapalainen, E. F., Cristovam, P., & Gomes, J. Ã. P.. (2008). Assessment of the use of cryopreserved x freeze-dried amniotic membrane (AM) for reconstruction of ocular surface in rabbit model. Arquivos Brasileiros De Oftalmologia, 71(5): 669-673.
  22. Liu, T., Liu, Y., Xie, L., He, X. & Ji Bai. (2013). Progress in the Pathogenesis of Pterygium. Current Eye Research 38 (12): 1191-1197. DOI: 10.3109/02713683.2013.823212
  23. Mahar PS and Manzar N. (2013). Pterygium recurrence related to its size and corneal involvement. Journal of the College of Physicians and Surgeons Pakistan 23(2):120-3.
  24. Marmamula, S., Khanna, C.K., Rao, G.N. (2013). Population-Based Assessment of Prevalence and Risk Factors for Pterygium in the South Indian State of Andhra Pradesh: The Andhra Pradesh Eye Disease Study. Investigative Ophthalmology & Visual Science 54(8):5359-5366.
  25. Mohamed, A., Sangwan, V.S. & Augusteyn, R.C. (2012). Growth of the human lens in the Indian adult population: Preliminary observations, Indian Journal of Ophthalmology 60 (6): 511–515. DOI: 10.4103/0301-4738.103775
  26. Nangia, V., Jonas, J. B., Nair, D., Saini, N., Nangia, P., & Panda-Jonas, S. (2013). Prevalence and Associated Factors for Pterygium in Rural Agrarian Central India. The Central India Eye and Medical Study. PLoS ONE, 8(12): 1-19.
  27. Park, Y. M., Kim, C. D., & Lee, J. S. (2015). Effect of Bevacizumab on Human Tenon's Fibroblasts Cultured from Primary and Recurrent Pterygium. The Korean Journal of Physiology & Pharmacology: Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology 19(4): 357-363.
  28. Qi, X., Zhang, D., Yuan, J., Yang, Z., Sun, Y., Wang, T., Ye, H., & Ling, Q. (2012). Relationship between angiogenesis and lymphangiogenesis in recurrent pterygium. International Journal of Ophthalmology 5(6): 655-660.
  29. Ribatti, D., Nico, B., Perra, M. T., Maxia, C., Piras, F., Murtas, D., Crivellato, E., & Sirigu, P. (2009). Correlation between NGF/TrkA and microvascular density in human pterygium. International Journal of Experimental Pathology 90(6): 615-620.
  30. Rodríguez-Ares, M.T., López-Valladares, M.J., Touriño, R., Vieites, B., Gude, F., Silva, M.T. and Couceiro. J. (2009). Effects of lyophilization on human amniotic membrane. Acta Ophthalmologica 87: 396-403.
  31. Sahin, S., Karabey, Y., Kaynak, M.S., Hincal, A.A. (2006). Potential Use of Freeze-Drying Technique for Estimation of Tissue Water Content, Methods Findinding Experimental and Clinical Pharmacology, 28(4): 211–215. DOI: 10.1358/mf.2006.28.4.990200
  32. Sato, Y., Ojima, H., Onaya, H., Mori, T., Hiraoka, N., Kishi, Y., Nara, S., Esaki, M., Shimada, K., Kosuge, T., Sugihara, K. and Kanai, Y. (2014), Hypervascular cholangiocellular carcinoma. Hepatology Research. 44: 1119-1129.
  33. Takahiro Nakamura, Makoto Yoshitani, Helen Rigby, Nigel J. Fullwood, Wakana Ito, Tsutomu Inatomi, Chie Sotozono, Tatsuo Nakamura, Yasuhiko Shimizu, Shigeru Kinoshita. (2004). Sterilized, Freeze-Dried Amniotic Membrane: A Useful Substrate for Ocular Surface Reconstruction. Investigative Ophthalmology & Visual Science 45(1):93-99. doi:
  34. Tan, D.T., Chee, S.P., Dear, K.B. & Lim, A.S. (1997). Effect of pterygium morphology on pterygium recurrence in a controlled trial comparing conjunctival autografting with bare sclera excision, Archive Ophthalmology 115(10):1235–1240.
  35. Thavarajah, R., Mudimbaimannar, V. K., Elizabeth, J., Rao, U. K., & Ranganathan, K. (2012). Chemical and physical basics of routine formaldehyde fixation. Journal of Oral and Maxillofacial Pathology: JOMFP, 16(3), 400-405.

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