Sains Malaysiana 48(9)(2019): 1927–1935

http://dx.doi.org/10.17576/jsm-2019-4809-14

Evaluation of the Human Amniotic Membrane as a Scaffold for Periodontal Ligament Fibroblast Attachment and Proliferation

(Penilaian Membran Amnion Manusia sebagai Perancah untuk Pelekatan dan Pengembangbiakan Fibroblas Ligamen Periodontal)

ASRAR ELAHI¹, HASLINA TAIB²*, ZURAIRAH BERAHIM², AHMAD AZLINA³ & SUZINA SHEIKH AB HAMID⁴

¹Department of Periodontics, College of Dentistry, Lahore Medical and Dental College, 53400, Lahore, Pakistan

²Periodontics Unit, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan Darul Naim, Malaysia

³Molecular Biology, School of Dental Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan Darul Naim, Malaysia

⁴Tissue Bank, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan Darul Naim, Malaysia

Received: 20 November 2018/Accepted: 13 June 2019

ABSTRACT

This study was aimed at evaluating the ability of the human amniotic membrane (HAM) to act as a scaffold for the growth of the main cells in periodontal regeneration, human periodontal ligament fibroblasts (HPDLFs). The HAM has many biological properties that are suitable for periodontal tissue regeneration such as low immunogenicity, anti-fibrosis, anti-inflammation, and a rich extracellular matrix component. Commercially available HPDLFs were seeded onto the HAM, and the attachment and proliferation of the cells were observed through scanning electron microscopy (SEM) and histological analysis. Cell viability was assessed using the alamarBlue® proliferation assay at days 1, 3, 7, 14 and 21. Histologically, the HPDLFs showed a monolayer to multilayer attachment onto the HAM from day 1 to day 7. The SEM analysis demonstrated that the HPDLFs had attached appropriately onto the HAM surface at day 1 to day 3, and began overlapping at day 7, while maintaining their flat shape. However, by days 14 and 21, there was an alteration in the morphology of the cells, where they later became rounded. The proliferation assay showed that the viability of the HPDLFs on the HAM had increased significantly from day 1 to day 7 (p=0.012), but later showed significant reduction at day 14 (p=0.002) and day 21 (p=0.005). In conclusion, this study showed that the HAM was able to function well as a scaffold for HPDLFs within 7 days, and thus, it can be a promising scaffold for periodontal regeneration. However, the behaviour of the cells in relation to the membrane over longer culture duration warrants further investigation.

Keywords: Amniotic membrane; periodontal fibroblasts; periodontal regeneration; scaffold

ABSTRAK

Kajian ini bertujuan untuk menilai keupayaan membran amniotik manusia (HAM) untuk bertindak sebagai perancah bagi pertumbuhan sel-sel utama dalam regenerasi periodontal iaitu sel fibroblas ligamen periodontal manusia (HPDLFs). HAM mempunyai banyak ciri biologi yang sesuai untuk pertumbuhan semula tisu periodontal seperti keimunogenan yang rendah, anti-fibrosis, anti-keradangan dan kaya dengan komponen matriks luar sel. HPDLFs yang tersedia secara komersial telah dibiakkan ke atas HAM, dan pelekatan serta pengembangbiakan HPDLFs diperhatikan melalui pemeriksaan mikroskop imbasan elektron (SEM) dan analisis histologi. Daya tahan sel telah dinilai menggunakan ujian percambahan alamarBlue® pada hari 1, 3, 7, 14 dan 21. Secara histologi, HPDLFs menunjukkan pelekatan ekalapisan kepada beberapa lapisan ke atas HAM daripada hari pertama hingga ke-7. Analisis SEM menunjukkan bahawa HPDLFs telah melekat dengan baik pada permukaan HAM pada hari pertama hingga hari ke-3 dan mula bertindih pada hari ke-7, sambil mengekalkan bentuknya yang rata. Bagaimanapun, pada hari ke-14 dan ke-21, terdapat perubahan pada morfologi sel, dengan mereka kemudiannya menjadi bulat. Ujian percambahan menunjukkan bahawa daya tahan sel HPDLFs pada HAM meningkat dengan ketara daripada hari pertama hingga hari ke-7 (p=0.012) namun kemudiannya menunjukkan penurunan yang ketara pada hari ke-14 (p=0.002) dan hari ke-21 (p=0.005). Kesimpulannya, kajian ini menunjukkan bahawa HAM dapat berfungsi dengan baik sebagai perancah untuk HPDLFs dalam tempoh 7 hari, dengan itu mampu menjadi perancah untuk regenerasi periodontal. Walau bagaimanapun, tingkah laku sel berhubung dengan membran ini dalam tempoh yang lebih lama masih memerlukan kajian lanjut.

Kata kunci: Fibroblas periodontal; membran amnion; perancah; pertumbuhan semula periodontal

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*Corresponding author; email: haslinakk@usm.my