Sains Malaysiana 51(4)(2022): 1131-1142

http://doi.org/10.17576/jsm-2022-5104-15

The Senescence Effect of Zoledronate on Three-Dimensional Oral Mucosa Model 

(Kesan Senesens Zoledronat pada Model Mukosa Oral Tiga Dimensi)

NUR BASHIRA SHAHARUDDIN¹, DANIEL JONES² & WEN LIN CHAI^1,*

¹Department of Restorative Dentistry, Faculty of Dentistry, University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

²Division of Natural Sciences, Indiana Wesleyan University, 4201 S. Washington St., Marion, Indiana, 46953, USA

Received: 18 October 2020/Accepted: 1 September 2021

Abstract

Zoledronate (ZOL) is an antiresorptive bisphosphonate used to prevent bone loss in skeletal-related disorders, especially in patients with advanced cancer metastatic to bone. However, this medication led to an oral lesion known as medication-related osteonecrosis of the jaw (MRONJ) which also presents clinically as unhealing soft tissue in the jaw. Keratinocytes are thought to be a significant onset factor for MRONJ and recent findings have shown evidence of senescence in keratinocytes. However, little is known about the effect of senescence-associated inflammation that may cause age-associated tissue degeneration, which relates mainly to the expression of extracellular modulators including cytokines known as senescence-associated secretory phenotype (SASP). The aim of this experiment was to investigate the effect of ZOL treatment and senescence-associated secretory phenotype (SASP) if any, on a three-dimensional oral mucosa model (OMM) in which a novel modification was made to reflect the existence of basement membrane (BM) and lamina propria accurately. The ZOL dosage for the model was optimized by exposing the immortalized human oral keratinocyte line (OKF 6/TERT-2) and normal human oral fibroblasts (NHOF) to ZOL at increasing dosages and then histoarchitecture of OMM was examined. Analysis of the model's histology showed significant epithelial thinning upon ZOL treatment and breakage of the BM accompanied by downward proliferation towards the lamina propria. A significant release of SASP molecules (MMP-3 and IL-8) was also detected upon treatment. Therefore, this study suggests that ZOL may impair healing at multiple types of tissues, originally from keratinocytes, by the deleterious effects of the senescence-associated inflammatory response.

Keywords: Oral mucosa; osteonecrosis; senescence; tissue engineering; zoledronate

Abstrak

Zoledronat (ZOL) adalah ubat bifosfonat antiresorptif yang digunakan untuk mencegah kehilangan tulang pada gangguan yang berkaitan dengan rangka, terutama pada pesakit yang mengalami barah tulang. Walau bagaimanapun, ubat ini sering menyebabkan lesi pada mulut yang dikenali sebagai osteonekrosis rahang (MRONJ) yang juga dikenali secara klinikal sebagai tisu lembut di rahang yang tidak dapat disembuhkan. Keratinosit dianggap sebagai faktor permulaan yang signifikan untuk MRONJ dan penemuan baru-baru ini telah menunjukkan bukti penuaan pada keratinosit. Walau bagaimanapun, kesan keradangan yang berkaitan dengan kemerosotan tisu berkaitan dengan usia, terutamanya pelahiran modulator ekstrasel termasuk sitokin dikenali sebagai fenotip sekretori yang berkaitan dengan penuaan (SASP). Tujuan penyelidikan ini adalah untuk mengkaji kesan rawatan ZOL dan fenotip sekretori yang berkaitan dengan penuaan (SASP) jika ada, pada model mukosa oral tiga dimensi (OMM) dengan pengubahsuaian baru dibuat untuk mencerminkan adanya membran dasar (BM) dan lamina propria dengan tepat. Dos ZOL untuk model itu dioptimumkan dengan mendedahkan keratinosit oral manusia abadi (OKF 6/TERT-2) dan fibroblas oral manusia normal (NHOF) kepada ZOL pada peningkatan dos dan kemudian struktur tisu OMM diperiksa. Analisis histologi model menunjukkan penipisan epitelium yang signifikan pada rawatan ZOL dan kerosakan BM disertai dengan pertumbuhan keratinosit ke arah lamina propria. Pelepasan molekul SASP yang signifikan (MMP-3 dan IL-8) juga dikesan semasa rawatan. Oleh itu, kajian ini menunjukkan bahawa ZOL kemungkinan besar boleh mengganggu penyembuhan pelbagai jenis tisu, yang bermula dari keratinosit melalui tindak balas keradangan yang berkaitan dengan penuaan.

Kata kunci: Kejuruteraan tisu; mukosa mulut; osteonekrosis; senesens; zoledronat

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^*Corresponding author; email: chaiwl@um.edu.my