Sains Malaysiana 54(2)(2025): 403-414

http://doi.org/10.17576/jsm-2025-5402-07

 

Pengesahan Kertas Penapis Whatman Gred 595 sebagai Kertas Serapan untuk Persampelan Cecair Krevis Gingiva

(Validation of Whatman Filter Paper Grade 595 as Absorption Paper for Gingival Crevicular Fluid Sampling)

 

LEE CHIE HOW¹, LEE XIAO QING², ROHAYA MEGAT ABDUL WAHAB³, FARINAWATI YAZID³, NUR ATMALIYA LUCHMAN⁴ & SHAHRUL HISHAM ZAINAL ARIFFIN^5,*

 

¹Klinik Pergigian Beserah, Jalan Beserah, Kampung Pelindung, 26100 Kuantan, Pahang, Malaysia
²Klinik Pergigian Setapak, Tingkat 1, Wisma Rampai, Blok Pasifik, Jalan 34/26, Taman Sri Rampai, 53300 Kuala Lumpur, Malaysia
³Jabatan Kesihatan Pergigian Keluarga, Fakulti Pergigian, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia
⁴CytoMed Therapeutics (Malaysia) Sdn. Bhd., Bandar Baru Permas Jaya, 81750 Johor Bahru, Johor, Malaysia
⁵Jabatan Sains Biologi dan Bioteknologi, Fakulti Sains dan Teknologi, Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

 

Received: 28 July 2024/Accepted: 22 October 2024

 

Abstrak

Pengesanan penanda biologi sebagai alat diagnostik telah dikaji secara meluas. Dentin Sialofosfoprotein (DSPP) yang biasanya ditemui di dalam cecair krevis gingiva (GCF) merupakan penanda biologi awal untuk pengesanan penyerapan akar gigi. Pada masa kini, kertas serapan Periopaper® adalah satu-satunya pilihan berkos tinggi yang membolehkan persampelan GCF. Antara pengganti Periopaper berkos rendah adalah kertas penapis Whatman® Gred 595 (W595). Oleh itu, kajian ini bertujuan mengesahkan kertas W595 sebagai pengganti bagi kertas Periopaper® dengan kos yang lebih rendah dan mudah diperoleh. Struktur kertas penapis W595 dikaji pada peringkat mikroskopik menggunakan Mikroskop Imbasan Elektron (SEM), diikuti dengan kualiti penyerapan menggunakan Periotron® 8000 dalam tiga selang masa iaitu T1 (0 s), T2 (15 s) dan T3 (30 s). Seterusnya, penentuan kuantiti penyerapan DSPP ditentukan oleh ELISA manakala ujian sitotoksik titisan sel MC-3T3 melalui pendekatan ujian MTT dan morfologi sel. Hasil menunjukkan bahawa struktur kedua-dua kertas adalah hampir serupa di bawah pemerhatian SEM sehingga pembesaran 200×, tetapi kertas penapis W595 sahaja menunjukkan struktur mikroporositi di bawah pembesaran 1000×. Kualiti penyerapan pula menunjukkan kedua-dua kertas berupaya menyerap GCF pada isi padu yang sama (p>0.05), serta tiada perbezaan yang signifikan (p>0.05) pada kuantifikasi jumlah DSPP melalui pendekatan ELISA. Ujian sitotoksisiti menggunakan titisan sel MC-3T3 menunjukkan kedua-dua kertas mempunyai populasi peratus sel hidup melebihi 50% (IC₅₀). Sel juga didapati mengekalkan morfologi yang sama serta tiada perbezaan yang signifikan (p>0.05) terhadap ketumpatan sel apabila kedua-dua kertas dikulturkan selama 72 jam. Kesimpulannya, kertas penapis W595 mempunyai mikrostruktur, kesan penyerapan GCF, potensi penyerapan molekul DSPP dan kesitotoksikan terhadap sel yang hampir sama dengan kertas Periopaper® dan boleh menggantikannya untuk pengambilan sampel GCF.

 

Kata kunci: Cecair krevis gingiva; Periopaper®; Whatman® Gred 595

 

Abstract

The detection of biomarkers as a diagnostic tool has been widely studied. Dentin sialophosphoprotein (DSPP), commonly found in gingival crevicular fluid (GCF), is an early biological marker for the detection of root resorption. Currently, Periopaper® is the only absorption paper at a high cost that allows the collection of GCF samples. Therefore, this study aims to validate Whatman® Filter Paper Grade 595 (W595) as a suitable substitution of Periopaper® for lower cost and easy access. The structure of W595 was tested by microscopic observation under a scanning electron microscope (SEM), followed by its absorbent quality using Periotron® 8000 over three different time intervals, T1 (0 s), T2 (15 s) and T3 (30 s). The DSPP absorption quantity was determined by enzyme-linked immunosorbent assay (ELISA) technique while the cytotoxic test of MC-3T3 cells was done by MTT and cell morphology test. Results showed that the structure of both papers were similar under 200× magnification but W595 paper had a higher microporosity under 1000× magnification. The absorption quality showed that both papers absorbed similar volumes of GCF (p>0.05), and there was no significant difference (p>0,05) in the amount of DSPP quantified from the sample. Cytotoxicity test using drops of MC-3T3 cells showed that both papers had a percentage population of living cells above 50% (IC₅₀). Cells maintained the same morphology and there was no significant difference (p>0.05) in cell density when cultured for 72 h. In conclusion, W595 paper has characteristics similar to those of Periopaper® paper and can be used as a substitute to collect samples of GCF. The W595 paper exhibits a microstructure, GCF absorption effects, DSPP molecule absorption potential, and cytotoxicity nearly identical to Periopaper®. Therefore, W595 paper can be a suitable replacement for Periopaper® in GCF sampling.

 

Keywords: Gingival crevicular fluid; Periopaper®; Whatman® Grade-595

 

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*Corresponding author; email: hisham@ukm.edu.my