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

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. 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, kaj...

Amniotic Membrane: An Approach to Periodontal Regeneration

“…Seven studies on GTR demonstrated that AM was an effective barrier able to enhance bone fill and improve periodontal parameters (probing pocket depth [PPD] and clinical attachment loss [CAL]). One laboratory study reported AM as a suitable scaffold for periodontal fibroblast cell growth [11]. The findings from all selected studies are summarized in Table 1.…”

“…Consequently, it is a readily available and cost-effective biomaterial for scaffolds in tissue engineering [10]. Scanning electron microscopy analysis of AM revealed rough surface architecture with the presence of microporosity, which may provide a suitable platform for cell attachment [11]. AM was used for wound treatment more than a century ago as a skin graft substitute for open wound for treating burnt and ulcerated skin surfaces by which it can accelerate epithelialization and reduce pain [12][13][14].…”

Amniotic Membrane as a Scaffold for Periodontal Tissue Engineering

“…40 Through the advancement in clinical and laboratory techniques, HAM is now being used as a membrane in guided tissue regenerative procedures, substrate for culturing stem cells and as scaffolds for delivering regenerative cells into oral defects (Table 4). 11,14…”

“…Nevertheless, with the current findings on the superiority of HAM histologically as well as clinically that are comparable with the current, available scaffolds for tissue engineering is a promising, alternative biomaterial in the management of periodontal defects. 14,22,36…”

“…13 The method for procurement and decellularization of HAM prior to seeding of specific cells has been reported. 14 This review is written with the intention of highlighting this biological membrane, which covers the histology of HAM, its medical and dental applications and the potential of HAM as a substrate or scaffold in periodontal tissue engineering. A computerized search was performed from databases; PubMed, Scopus, and Google Scholar, by using the keywords: "amniotic membrane", "human amniotic membrane", "periodontal regeneration", "periodontal tissue engineering", "scaffold" and "scaffold in periodontal regeneration".…”