Studi in Vitro Uji Sitotoksisitas Gel Asap Cair Tempurung Kelapa terhadap Sel Ligamen Periodontal sebagai Bahan Perawatan Saluran Akar dengan Metode MTT Assay
DOI:
https://doi.org/10.36312/mjketr61Keywords:
Gel Asap Cair Tempurung Kelapa, Sitotoksisitas, Sel Ligamen Periodontal, MTT Assay, BiokompatibilitasAbstract
Bahan irigasi yang saat ini banyak digunakan, seperti natrium hipoklorit (NaOCl), klorheksidin, dan EDTA, terbukti efektif menghilangkan bakteri namun memiliki risiko toksisitas terhadap sel dan jaringan sekitarnya. Paparan NaOCl pada konsentrasi tinggi dapat menyebabkan iritasi jaringan, nekrosis, hingga reaksi ekstrusi yang serius pada jaringan periapical. Penelitian ini bertujuan untuk mengevaluasi sitotoksisitas gel asap cair tempurung kelapa (CS-LS) terhadap sel ligamen periodontal (PDL cells) sebagai kandidat bahan perawatan saluran akar menggunakan metode MTT assay. Penelitian eksperimental in vitro dengan rancangan pre-test dan post-test group design melibatkan tujuh kelompok perlakuan (kontrol media, kontrol sel, kontrol positif NaOCl 5,25%, serta gel CS-LS pada konsentrasi 6,25%, 12,5%, 50%, 75%, dan 100%) dengan lima replikasi per kelompok. Viabilitas sel dihitung berdasarkan nilai absorbansi MTT (= 570 nm) dan dianalisis secara statistik menggunakan uji Kruskal–Wallis diikuti uji Mann–Whitney bila diperlukan. Hasil menunjukkan perbedaan bermakna secara statistik pada nilai absorbansi antar kelompok (p<0,05). Persentase viabilitas sel tertinggi tercatat pada gel CS-LS 6,25% (76,30%), sehingga memenuhi kriteria biokompatibilitas (viabilitas >70% menurut ISO10993-5), sedangkan pada konsentrasi 12,5% viabilitas menurun menjadi sekitar 50,25% (mendekati nilai IC50). Konsentrasi tinggi (50–100%) menghasilkan penurunan viabilitas yang substansial (31–33%), meskipun masih menunjukkan viabilitas relatif lebih baik dibandingkan NaOCl 5,25%. Berdasarkan temuan ini, gel CS-LS pada konsentrasi di bawah 12,5% tampak aman dan berpotensi dikembangkan sebagai alternatif bahan irigasi endodontik; namun penelitian lanjutan yang memperluas rentang konsentrasi, menambahkan parameter biologis (misalnya: proliferasi, ekspresi gen osteogenik, marker inflamasi), serta studi in vivo dan uji biokompatibilitas jangka panjang direkomendasikan untuk mengonfirmasi keamanan dan potensi aplikasi klinis.
In Vitro Study of Cytotoxicity Test of Liquid Coconut Shell Smoke Gel on Periodontal Ligament Cells as Root Canal Treatment Material Using MTT Assay Method
Abstract
Currently widely used irrigants, such as sodium hypochlorite (NaOCl), chlorhexidine, and EDTA, have been shown to be effective in eliminating bacteria but carry the risk of toxicity to surrounding cells and tissues. Exposure to NaOCl at high concentrations can cause tissue irritation, necrosis, and even serious extrusion reactions in periapical tissues. This study aimed to evaluate the cytotoxicity of coconut shell liquid smoke gel (CS-LS) on periodontal ligament (PDL) cells as a potential root canal treatment material using the MTT assay method.
This research was an in vitro experimental study with a pre-test and post-test group design, involving seven treatment groups: media control, cell control, positive control (5.25% NaOCl), and CS-LS gel at concentrations of 6.25%, 12.5%, 50%, 75%, and 100%, with five replicates in each group. Cell viability was determined based on MTT absorbance values (= 570 nm) and statistically analyzed using the Kruskal–Wallis test followed by the Mann–Whitney post hoc test when appropriate. The results demonstrated a statistically significant difference in absorbance values among the treatment groups (p < 0.05). The highest cell viability was observed in the 6.25% CS-LS gel group (76.30%), meeting the biocompatibility criteria (cell viability > 70% according to ISO 10993-5), whereas the 12.5% concentration showed a reduced viability of approximately 50.25%, approaching the IC50 value. Higher concentrations (50–100%) resulted in a substantial decrease in cell viability (approximately 31–33%), although these values remained relatively higher than those observed with 5.25% NaOCl. In conclusion, CS-LS gel at concentrations below 12.5% exhibits low cytotoxicity and potential for development as an alternative endodontic irrigant; however, further studies incorporating a wider range of concentrations, additional biological parameters (such as cell proliferation, osteogenic gene expression, and inflammatory markers), as well as in vivo and long-term biocompatibility assessments, are necessary to confirm its safety and clinical applicability.
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