Azalea Nur-Qistina Azami1, Khairani Idah Mokhtar2*, Basma Ezzat Mustafa2, Norzaity Mohd. Kenali3, Munirah Sha’ban1, Azlina Ahmad4, Ismail Zainol5
1. Department of Physical Rehabilitation Sciences, Kulliyyah of Allied Health Sciences, International Islamic University Malaysia, Kuantan Campus, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia
2. Department of Fundamental Dental and Medical Sciences, Kulliyyah of Dentistry, International Islamic University Malaysia, Kuantan Campus, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia
3. Department of Pediatrics Dentistry, Kulliyyah of Dentistry, International Islamic University Malaysia, Kuantan Campus, Bandar Indera Mahkota, 25200 Kuantan, Pahang, Malaysia
4. Basic and Medical Sciences Unit, School of Dental Sciences, Universiti Sains Malaysia, Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia
5. Faculty of Science and Mathematics, Universiti Pendidikan Sultan Idris, 35900 Tanjong Malim, Perak, Malaysia
*Corresponding author: Professor Dr Khairani Idah Mokhtar.
ABSTRACT
INTRODUCTION: Tanduk rusa, or deer velvet antlers (DVA), has long been utilised in traditional Asian medicine for its various health benefits, including bone health maintenance and regeneration. Nonetheless, the fundamentals of scientific merit are still limited.
OBJECTIVE(S): This study investigates the effects of DVA extract on stem cells derived from human exfoliated deciduous teeth (SHED); capable of differentiating into osteoblasts, which are essential for bone formation and healing. To achieve this, the effect of DVA on the biological activity of SHED and its potential to differentiate into osteoblast-like cells were determined accordingly.
MATERIALS & METHODS: Powdered form of DVA obtained from male Malayan deer (Cervus timorensis) was extracted using sterile water, and its impact on SHED was assessed at different concentrations (0 to 200 µg/ml). Cell viability was determined using MTT assay, cell proliferation using AlamarBlue assay while population doubling time (PDT) was determined according to the available protocol, and morphology was observed under the inverted microscope. Osteoblast differentiation was analyzed by measuring alkaline phosphatase (ALP) activity, calcium deposition via Alizarin Red-S staining, and expression of osteoblast markers (RUNX2, OSX, and OCN) in SHED cultured in osteoblast differentiation medium (ODM) were analysed using RT-qPCR. Statistical analyses were performed using One-way ANOVA and post hoc Tukey’s test with p<0.05 is considered as significant.
RESULTS: DVA extract was non-toxic to SHED at all tested concentrations, with 3.125 µg/ml identified as the optimal concentration for promoting proliferation and reducing PDT. Morphologically, SHED maintained a fibroblast-like shape at all concentrations analysed. When cultured in ODM, addition of DVA resulted in an increase of ALP activity at Day 21 (P<0.05), strong calcium mineralization at Day 14 onwards, and higher expression of osteoblast markers (RUNX2, OCN). Changes in morphology into osteoblast-like was observed.
CONCLUSION: DVA stimulates osteoblast differentiation of SHED, particularly during mineralization phases, highlighting its potential as a natural bone development stimulant.
KEYWORDS: Deer Velvet Antler, Tanduk Rusa, SHED, Osteoblast Differentiation
