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Fulltext Effects of different processing methods of human amniotic membrane on the quality of extracted RNA

Rusidah Mat Yatim, Kannan, Thirumulu Ponnuraj, Suzina Sheikh Ab Hamid, Shazana Hilda Shamsudin

Archives of Orofacial Sciences, 2013;8(2):47-53.
MyJurnal

The aim of this study was to determine the efficiency of different human amniotic membrane (HAM) processing methods on the concentration, purity and integrity of RNA. Two different techniques (Technique1 andTechnique2) were employed for the processing of HAM, which differed in terms of washing solution, sample storage conditions and processing time. Based on preservation of HAM, three groups were formed under each technique. In Technique 1, the groups were fresh frozen 1 (F1), glycerol preserved (GP) and gamma irradiated glycerol preserved (IGP); where else in Technique 2, the groups were fresh frozen 2 (F2), 50%glycerol/Dulbecco’s modified Eagle medium (DMEM) cryopre served HAM diluted with phosphate buffered saline(GB) and 50% glycerol/DMEM cryop reserved HAM diluted with diethyl procarbonate water (GD). Total RNA was extracted from the samples and their concentration, purity and integrity were examined. The F2 sample of which there was no pre-washing step and involved direct sample storage at-80oC, shorter processing time and chilled processing conditions had yielded better quality of RNA compared to the others.
Fulltext In vitro cytotoxicity evaluation of dental porcelain using human cell lines

Hazem Yousef Abu Sharbeh, Kannan, Thirumulu Ponnuraj, Raja Azman Raja Awang, Adam Husein

Archives of Orofacial Sciences, 2012;7(2):68-74.
MyJurnal

The in vitro cytotoxic potential of locally produced dental porcelain was evaluated in this study. The cellular response of human osteoblast and fibroblast cell lines were assessed using MTT assay by incubating with the fluid extract of dental porcelain powder and dental porcelain discs (direct test). Aging process was carried out by submerging the discs into 3% bovine serum albumin (BSA) solution. Tests on extracts showed that dental porcelain was significantly different from the control at a concentration of 250 mg/ml. Direct test showed that dental porcelain after aging was not significantly different from the control with a mean (SD) of 89.2 (13.4)%, whereas, it was significantly different from the control before conditioning of BSA with a mean (SD) of 88.5 (12.1)%. However, the dental porcelain caused mild suppression of succinate dehydrogenase activity (
Fulltext Relevance of BRCA1 and BRCA2 variants in treating triple negative breast cancer patients

Engku Fatimah Syairah Engku Safruddin, Wan Zainira Wan Zain, Bhavaraju, Venkata Murali Krishna, Kannan, Thirumulu Ponnuraj

Archives of Orofacial Sciences, 2017;12(2):69-76.
MyJurnal

Given that the germline mutations of BRCA1 and BRCA2 confer genetic susceptibility to cancer, the
genetic variations, polymorphisms or mutations are widely analyzed in Western countries. However, in Asian
population, the prevalence of BRCA1 and BRCA2 polymorphisms is very limited. In Asia, breast cancer occurs in
women early with an age of onset under 50 years. This review comprises the incidence of BRCA1 and BRCA2
polymorphisms in the Japanese, Korean and Malaysian population. Founder mutations of BRCA1 and BRCA2
were also compared to mark the genetic difference in these populations. The mutational analysis performed to
analyze the entire coding region of BRCA1 and BRCA2 include the next generation sequencing and full
sequencing of all exons and intron-exon junctions. From the diagnosis of triple negative breast cancer (TNBC)
patients, TNBC is associated with the lack of tailored therapies and the treatment option available for TNBC
patients is mainly chemotherapy. The poor prognosis of TNBC leads to determine the predictive biomarkers in
order to develop treatment efficacy. This review will address the current clinical therapies available to treat TNBC
patients.
Fulltext Genotoxic evaluation of synthetic hydroxyapatite using mammalian bone marrow chromosome aberration test

Kannan, Thirumulu Ponnuraj, Nik Ahmad Shah Nik Lah, Azlina Ahmad, Siti Fatimah Ramli, Narazah Mohd Yusof, Ab Rani Samsudin

Archives of Orofacial Sciences, 2014;9(1):1-7.
MyJurnal

Some of the beneficial bio compatible properties of hydroxyapatite [Ca10(PO4)6(OH)2]; the major componentand an essential ingredient of normal bone and teeth, are that it is rapidly integrated into the human body and will bondto bone forming in distinguishable unions. But, before new materials are approved for medical use, mutagenesis systems to exclude cytotoxic, mutagenic or carcinogenic properties are applied worldwide. This study aimed to detectany chromosomal aberrations induced by the synthetic hydroxyapatite granules [Manufactured by Universiti Sains
Malaysia, (USM) Penang, Malaysia] in the bone marrow cells of mice. The mitotic indices of the groups treated with synthetic hydroxya patite granules did not show any significant difference as compared to the negative control group treated with distilled water. Also the groups of mice treated with synthetic hydroxyapatite granules and distilled waterdid not induce significant change in chromosome aberrations as compared to the positive control group treated with Mitomycin C. The mitotic indices and chromosomal analyses indicate that under the present test conditions, synthetichydroxya patite granules (manufactured by USM) are non cytotoxic and do not induce chromosome aberrations in the bone marrow cells of mice.
Fulltext Genotoxicity assessment of locally produced dental nanocomposite using Comet assay

Siti Robayah Mohd Zakri, Kannan, Thirumulu Ponnuraj, Nora Aziz, Siti Fadilah Abdullah, Dasmawati Mohamad, Ismail Ab Rahman, et al.

Archives of Orofacial Sciences, 2011;6(1):15-20.
MyJurnal

The aim of this study was to determine the genotoxicity of a locally produced nanocomposite by Universiti Sains Malaysia, Malaysia using Comet assay. Stem cells from human exfoliated deciduous teeth (SHED) were treated with the nanocomposite at five different concentrations (0.006, 0.0125, 0.025, 0.05, and 0.1 mg/ml) along with concurrent negative (medium alone) and positive control (zinc sulfate heptahydrate) and incubated at 37°C for 24 hours in an incubator at 5% CO2. The tail moment was used to assess the extent of DNA damage. The tail moment for the group of SHED treated with nanocomposite (for all the five different concentrations) was not statistically significant as compared to the negative control, suggesting that the locally produced dental nanocomposite did not induce any DNA damage. Hence, it can be concluded that the locally produced nanocomposite is non-genotoxic on stem cells from human exfoliated deciduous teeth.
Chemical constituents of Malaysian apis mellifera propolis

Tuan Nadrah Naim Tuan Ismail, Kannan Thirumulu Ponnuraj, Siti Amrah Sulaiman, Kannan Thirumulu Ponnuraj, Che Nin Man, Norul Badriah Hassan

Sains Malaysiana, 2018;47:117-122.

Propolis is a resinous material collected by honey bees from various plant sources. The chemical constituents differ depending on the source of vegetation area. To date, no study of the chemical constituents of Apis mellifera propolis from Malaysia has been reported. Thus, the aim of this study was to determine the chemical constituents of Malaysian Apis mellifera propolis. Raw propolis samples were collected from Acacia mangium apiary from northern and southern regions of Peninsular Malaysia and were extracted using 70% ethanol. Then, the ethanolic extracts were evaporated and were lyophilized to dryness. The silylated compounds were identified by gas chromatography with mass spectrometry analysis (GC-MS). The compounds were characterised by comparison with NIST02 and Wiley275 libraries. Twenty-six individual compounds were identified and β-panasinsene was identified for the first time in propolis. The main compounds identified from both southern and northern regions of Peninsular Malaysia were terpenoids. Terpenoids belong to an important class in the propolis compounds due to its valuable biological properties. The amount of terpenoids ranged from 7.19 to 11.46% indicating that although the main plant source for both regions was Acacia mangium, other shrubs and fruit orchards surrounding the two apiaries may differ which may contribute to the variations.
Fulltext Cytogenetics: past, present and future

Kannan TP, Zilfalil BA

Malays J Med Sci, 2009 Apr;16(2):4-9.
PMID: 22589651 MyJurnal

Fifty years have elapsed since the discovery of the number of human chromosomes in 1956. Newer techniques have been developed since then, ranging from the initial conventional banding techniques to the currently used molecular array comparative genomic hybridisation. With a combination of these conventional and molecular techniques, cytogenetics has become an indispensable tool for the diagnosis of various genetic disorders, paving the way for possible treatment and management. This paper traces the history and evolution of cytogenetics leading up to the current state of technology.
Fulltext Keloid scarring: understanding the genetic basis, advances, and prospects

Halim AS, Emami A, Salahshourifar I, Kannan TP

Arch Plast Surg, 2012 May;39(3):184-9.
PMID: 22783524 DOI: 10.5999/aps.2012.39.3.184

Keloid disease is a fibroproliferative dermal tumor with an unknown etiology that occurs after a skin injury in genetically susceptible individuals. Increased familial aggregation, a higher prevalence in certain races, parallelism in identical twins, and alteration in gene expression all favor a remarkable genetic contribution to keloid pathology. It seems that the environment triggers the disease in genetically susceptible individuals. Several genes have been implicated in the etiology of keloid disease, but no single gene mutation has thus far been found to be responsible. Therefore, a combination of methods such as association, gene-gene interaction, epigenetics, linkage, gene expression, and protein analysis should be applied to determine keloid etiology.
Genotoxicity evaluation of locally produced dental porcelain--an in vitro study using the Ames and Comet assays

Noushad M, Kannan TP, Husein A, Abdullah H, Ismail AR

Toxicol In Vitro, 2009 Sep;23(6):1145-50.
PMID: 19505568 DOI: 10.1016/j.tiv.2009.05.025

The aim of this study was to determine the genotoxicity of a locally produced dental porcelain (Universiti Sains Malaysia, Malaysia) using the Ames and Comet assays. In the Ames assay, four genotypic variants of the Salmonella strains (TA98, TA100, TA1537 and TA1535) carrying mutations in several genes were used. The dental porcelain was incubated with these four strains in five different doses both in the presence and absence of metabolic activation (S9) and the result was assessed based on the number of revertant colonies. Concurrently, appropriate positive controls were used so as to validate the test. The average number of revertant colonies per plate treated with locally produced dental porcelain was less than double as compared to that of negative control. In the Comet assay, L929 (CCL-1 ATCC, USA) mouse fibroblast cells were treated with the dental porcelain in three different concentrations along with concurrent negative and positive controls. The tail moment which was used as a measurement of DNA damage was almost equal to that of the negative control, suggesting that the locally produced dental porcelain did not induce any DNA damage. The results indicated that the locally produced dental porcelain is non-genotoxic under the present test conditions.
In vitro genotoxicity tests for polyhydroxybutyrate--a synthetic biomaterial

Ali AQ, Kannan TP, Ahmad A, Samsudin AR

Toxicol In Vitro, 2008 Feb;22(1):57-67.
PMID: 17892925

The aims of this study are to determine the mutagenicity of a locally produced polyhydroxybutyrate (PHB) using Salmonella mutagenicity test and to find out if PHB altered the expression of p53 and c-myc proto-oncogenes and bcl-xl and bcl-xs anti-apoptotic genes in the human fibroblast cell line, MRC-5. Different concentrations of PHB were incubated with special genotypic variants of Salmonella strains (TA1535, TA1537, TA1538, TA98 and TA100) carrying mutations in several genes both with and without metabolic activation (S9) and the test was assessed based on the number of revertant colonies. The average number of revertant colonies per plate treated with PHB was less than double as compared to that of negative control. For the gene expression analyses, fibroblast cell lines were treated with PHB at different concentrations and incubated for 1, 12, 24 and 48 h separately. The total RNA was isolated and analysed for the expression of p53, c-myc, bcl-xl and bcl-xs genes. The PHB did not show over or under expression of the genes studied. The above tests indicate that the locally produced PHB is non-genotoxic and does not alter the expression of the proto-oncogenes and anti-apoptotic genes considered in this study.
Fulltext Biological Interaction Between Human Gingival Fibroblasts and Vascular Endothelial Cells for Angiogenesis: A Co-culture Perspective

Um Min Allah N, Berahim Z, Ahmad A, Kannan TP

Tissue Eng Regen Med, 2017 Oct;14(5):495-505.
PMID: 30603504 DOI: 10.1007/s13770-017-0065-y

Advancement in cell culture protocols, multidisciplinary research approach, and the need of clinical implication to reconstruct damaged or diseased tissues has led to the establishment of three-dimensional (3D) test systems for regeneration and repair. Regenerative therapies, including dental tissue engineering, have been pursued as a new prospect to repair and rebuild the diseased/lost oral tissues. Interactions between the different cell types, growth factors, and extracellular matrix components involved in angiogenesis are vital in the mechanisms of new vessel formation for tissue regeneration. In vitro pre-vascularization is one of the leading scopes in the tissue-engineering field. Vascularization strategies that are associated with co-culture systems have proved that there is communication between different cell types with mutual beneficial effects in vascularization and tissue regeneration in two-dimensional or 3D cultures. Endothelial cells with different cell populations, including osteoblasts, smooth muscle cells, and fibroblasts in a co-culture have shown their ability to advocate pre-vascularization. In this review, a co-culture perspective of human gingival fibroblasts and vascular endothelial cells is discussed with the main focus on vascularization and future perspective of this model in regeneration and repair.
Evaluation of Tualang honey as a supplement to fetal bovine serum in cell culture

Kannan TP, Ali AQ, Abdullah SF, Ahmad A

Food Chem Toxicol, 2009 Jul;47(7):1696-702.
PMID: 19394390 DOI: 10.1016/j.fct.2009.04.020

The aim of this study was to evaluate Tualang honey as a supplement to fetal bovine serum in cell cultures using MTT assay, chromosome aberration test and gene expression analyses. The MTT assay showed the highest percentage of cell proliferation (105.3% increment than control) of human osteoblast cell line (CRL 1543) in 0.0195% honey in Dulbecco's modified eagle medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. There was enhanced cell proliferation corresponding to the decrease in concentrations of honey as indicated by the mitotic index values when the osteoblast cell line was incubated at 37 degrees C for 48 hours. There were no chromosome aberrations both in the honey treated as well as distilled water treated (negative control) cell lines. In the case of gene expression analyses, fibroblast cell lines (CCL 171) were treated with honey (0.0195%) for 24 and 48 hours separately. Though there was over expression for the bcl-xl gene at both 24 and 48 hours, under expression for bcl-xs gene at 24 hours and over expression at 48 hours and under expression for both c-myc and p53 genes at both 24 and 48 hours, none of them were statistically significant in altering the expression of mRNA.
Identification of Copy Number Variation Among Nonsyndromic Cleft Lip and or Without Cleft Palate With Hypodontia: A Genome-Wide Association Study

Ghazali N, Abd Rahman N, Ahmad A, Sulong S, Kannan TP

Front Physiol, 2021;12:637306.
PMID: 33732167 DOI: 10.3389/fphys.2021.637306

Nonsyndromic cleft lip and or without cleft palate (NSCL/P) with the hypodontia is a common developmental abnormality in humans and animals. This study identified the genetic aberration involved in both NSCL/P and hypodontia pathogenesis. A cross-sectional study using genome-wide study copy number variation-targeted CytoScan 750K array carried out on salivary samples from 61 NSCL/P and 20 noncleft with and without hypodontia Malay subjects aged 7-13 years old. Copy number variations (CNVs) of SKI and fragile histidine triad (FHIT) were identified in NSCL/P and noncleft children using quantitative polymerase chain reaction (qPCR) as a validation analysis. Copy number calculated (CNC) for each gene determined with Applied Biosystems CopyCaller Software v2.0. The six significant CNVs included gains (12q14.3, 15q26.3, 1p36.32, and 1p36.33) and losses (3p14.2 and 4q13.2) in NSCL/P with hypodontia patients compared with the NSCL/P only. The genes located in these regions encoded LEMD3, IGF1R, TP73, SKI, FHIT, and UGT2β15. There were a significant gain and loss of both SKI and FHIT copy number in NSCL/P with hypodontia compared with the noncleft group (p < 0.05). The results supported that CNVs significantly furnish to the development of NSCL/P with hypodontia.
Screening of Transforming Growth Factor Beta 3 and Jagged2 Genes in the Malay Population With Nonsyndromic Cleft Lip With or Without Cleft Palate

Ghazali N, Rahman NA, Kannan TP, Jaafar S

Cleft Palate Craniofac J, 2015 07;52(4):e88-94.
PMID: 26151095 DOI: 10.1597/14-024

OBJECTIVE: To determine the prevalence of mutations in transforming growth factor beta 3 (TGFβ3) and Jagged2 genes and their association with nonsyndromic cleft lip with or without cleft palate (CL±P) patients.

DESIGN: Cross-sectional study on nonsyndromic CL±P and noncleft patients.

SETTING: Reconstructive clinic and outpatient dental clinic, Hospital Universiti Sains Malaysia.

PATIENTS: Blood samples of 96 nonsyndromic CL±P and 96 noncleft subjects.

MAIN OUTCOME MEASURE: Prevalence and association of mutations in TGFβ3 and Jagged2 genes with nonsyndromic CL±P.

RESULTS: Most of the nonsyndromic CL±P patients (53.1%) had left unilateral CLP. There were slightly more females (56.6%) compared with males. The prevalence of the mutations in the TGFβ3 gene was 17.7% (95% confidence interval [CI]: 9.5, 24.5) and in the Jagged2 gene was 12.5% (95% CI: 5.5, 18.5), which was higher compared with the noncleft group. For the TGFβ3 gene, there was no mutation in the coding region in either of the groups. All variants were single nucleotide polymorphisms located within the intronic flanking region. Two variants were identified (g.15812T>G and g.15966A>G) in both nonsyndromic CL±P and noncleft patients. However, the association was not significant (P > .05). Three variants (g.19779C>T, g.19547G>A, and g.19712C>T) were identified in the Jagged2 gene among nonsyndromic CL±P and noncleft patients. Only g.19712C>T showed a significant association with nonsyndromic CL±P patients (P = .039).

CONCLUSION: g.19712C>T might play a crucial role in the development of cleft lip and palate. To the best of our knowledge, this is the first report of the mutation found within intron 13 of the Jagged2 gene among nonsyndromic CL±P Malay patients.

Study site:Reconstructive and outpatient dental clinic, Hospital Universiti Sains Malaysia (HUSM)
Effect of gamma radiation on the expression of mRNA growth factors in glycerol cryopreserved human amniotic membrane

Yatim RM, Kannan TP, Ab Hamid SS

Cell Tissue Bank, 2016 Dec;17(4):643-651.
PMID: 27535136

Human amniotic membrane (HAM) due to its high biocompatibility, low immunogenicity, anti-microbial, anti-viral properties as well as the presence of growth factors has been used in various clinical applications. The growth factors play an important role in wound healing. The current study aimed to explore the effect of 15 kGy gamma radiation dose on selected growth factors and receptors mRNA present in HAM. Eight growth factors, namely, EGF, HGF, KGF, TGF-α, TGF-β1, TGF-β2, TGF-β3 and bFGF and two growth factor receptors, HGFR and KGFR were evaluated in this study. The total RNA was extracted and converted to complimentary DNA using commercial kits. Subsequently, the mRNA expressions of these growth factors were evaluated using real-time PCR and the results were statistically analyzed using REST-MCS software. This study confirmed the presence of these mRNA growth factors and receptors in fresh, glycerol cryopreserved and irradiated glycerol cryopreserved HAM. In glycerol cryopreserved HAM, the results showed up-regulation of HGF and bFGF and down-regulation of EGF, HGFR, KGF, KGFR, TGF-α, TGF-β1, TGF-β2 and TGF-β3 relative to the fresh HAM which acted as the control, whereas in irradiated glycerol cryopreserved HAM, the results showed up-regulation of EGF, HGF, KGF, KGFR, TGF-β1, TGF-β2 and TGF-β3 and down-regulation of HGFR, TGF-α and bFGF relative to the glycerol cryopreserved HAM which acted as the control. However, these mRNA expressions did not show any statistical significant difference compared to the control groups. This study concluded that a dose of 15 kGy of gamma radiation did not affect the mRNA expression for the growth factors' and receptors' in the glycerol cryopreserved HAM.
Properties of Cell Sources in Tissue-Engineered Three-dimensional Oral Mucosa Model: A Review

Mohd Nor NH, Berahim Z, Ahmad A, Kannan TP

Curr Stem Cell Res Ther, 2017;12(1):52-60.
PMID: 27538403

Oral mucosa is a mucous membrane lining the oral cavity. Its main function is to protect the deeper structures against the external factors; thermal, chemical, mechanical and biological stimuli. Apart from that, it also plays a significant role during mastication, deglutition and speech. Some oral diseases or injuries to oral mucosa lead to impairment of the oral functions and aesthetics which eventually result in permanent defect of oral mucosa. In order to overcome this defect, different approaches for the development of reconstructed oral mucosa models have been employed including skin/autologous grafts, guided tissue replacement, vestibuloplasty etc. However, the finding of an acceptable source for the transplantations or autologous grafts seems a bit challenging. To overcome this problem, the development of oral mucosa using tissue engineering approach has been widely studied involving various cell lines from different sources. This paper aims to highlight various cell sources used in the development of tissueengineered oral mucosa models based on articles retrieved from PubMed and MEDLINE databases using the search terms "oral mucosa tissue engineering", regardless of time when published.
The synergistic effects of IL-6/IL-17A promote osteogenic differentiation by improving OPG/RANKL ratio and adhesion of MC3T3-E1 cells on hydroxyapatite

Sritharan S, Kannan TP, Norazmi MN, Nurul AA

J Craniomaxillofac Surg, 2018 Aug;46(8):1361-1367.
PMID: 29805067 DOI: 10.1016/j.jcms.2018.05.002

OBJECTIVE: In this study, we evaluated the potential role of IL-6 and/or IL-17A in regulating the OPG/RANKL (osteoprotegerin/receptor activator of nuclear factor kappa b ligand) system of murine osteoblast cell line (MC3T3-E1) cultured on hydroxyapatite (HA).
METHODS: MC3T3-E1 cells were seeded on HA and treated with recombinant IL-6 or rIL-17A or combination of the two cytokines. Cell proliferation and differentiation activity were measured by MTS and alkaline phosphatase assays respectively. Observation of cell adhesion and proliferation was examined by scanning electron microscopy. Gene and protein expressions were performed on RANKL and OPG using qPCR, Western blot and ELISA.
RESULTS: We demonstrated that treatment with recombinant IL-17A (rIL-17A) and the combination rIL-6/rIL-17A promoted better adhesion and higher proliferation of cells on HA. Cells treated with rIL-17A and the combination cytokines showed a significant increase in differentiation activity on day 7, 10 and 14 as indicated by ALP activity (p
Interleukin-17A promotes osteogenic differentiation by increasing OPG/RANKL ratio in stem cells from human exfoliated deciduous teeth (SHED)

Sebastian AA, Kannan TP, Norazmi MN, Nurul AA

J Tissue Eng Regen Med, 2018 08;12(8):1856-1866.
PMID: 29774992 DOI: 10.1002/term.2706

Stem cells derived from human exfoliated deciduous teeth (SHED) represent a promising cell source for bone tissue regeneration. This study evaluated the effects of interleukin-17A (IL-17A) on the osteogenic differentiation of SHED. SHED were cultured in complete alpha minimum essential medium supplemented with osteoinducing reagents and treated with recombinant IL-17A. The cells were quantitatively analysed for proliferative activity by MTS assay, cell markers expression, and apoptotic activity by flow cytometry. For osteogenic differentiation, alkaline phosphatase (ALP) activity was quantified; mineralization assays were carried out using von Kossa and Alizarin red, and expression of osteogenic markers were analysed by real-time polymerase chain reaction and Western blot. The results showed that treatment with IL-17A increased proliferative activity in a dose-dependent manner, but reduced the expression of stem cell markers (c-Myc and Nanog) as the days progressed. IL-17A induced osteogenic differentiation in SHED as evidenced by high ALP activity, increased matrix mineralization, and upregulation of the mRNA expression of the osteogenic markers ALP, alpha 1 type 1 collagen (Col1A1), runt-related transcription factor 2 (RUNX2), osteopontin (OPN), osteocalcin (OCN), and osteoprotegerin (OPG) but downregulation of receptor activator of nuclear factor κB ligand (RANKL) as well as altering the OPG/RANKL ratio. Findings from our study indicate that IL-17A enhances proliferation and osteogenic differentiation of SHED by regulating OPG/RANKL mechanism thus suggests therapeutic potential of IL-17A in bone regeneration.
Fulltext Study on flaxseed crude extract on stem cells from human exfoliated deciduous teeth (SHED)

Nur Sazwi Nordin, Lokman Mohammad Isa, Syed Zahir Idid, Widya Lestari, Basma Ezzat Mustafa, Solachuddin Jauhari Arief Ikhwan, et al.

International Medical Journal Malaysia, 2017;16(21):48-48.
MyJurnal

Flaxseeds offer a wide range of pharmacological properties including antioxidant,
antibacterial and anticancer. However its effect on mesenchymal stem cells has not been
elucidated. Thus, this study aimed to determine the effects of flaxseed crude extract on stem cell
from human exfoliated deciduous teeth (SHED) in terms of cell viability, morphology and
proliferation activity. (Copied from article).
Identification of novel fibroblast-like cells from stem cells from human exfoliated deciduous teeth

Mohd Nor NH, Berahim Z, Azlina A, Kannan TP

Clin Oral Investig, 2019 Nov;23(11):3959-3966.
PMID: 30847574 DOI: 10.1007/s00784-019-02827-x

OBJECTIVES: This study aimed to differentiate and characterize fibroblast-like cells from stem cells from human exfoliated deciduous teeth (SHED).
MATERIALS AND METHODS: The differentiation of fibroblast-like cells from SHED was carried out by using specific human recombinant connective tissue growth factor (CTGF). To characterize fibroblastic differentiation, the induced cells were subjected to morphological changes, proliferation rate, gene expression analysis using quantitative reverse transcription-polymerase chain reaction (qRT-PCR), flow cytometry, and immunofluorescence staining. The commercial primary human gingival fibroblasts served as positive control in this study.
RESULTS: The results from characterization analysis were compared with that of commercial cells to ensure that the cells differentiated from SHED were fibroblast-like cells. The results showed the inductive effect of CTGF for fibroblastic differentiation in SHED. SHED-derived fibroblasts were successfully characterized despite having similar morphological appearance, i.e., (i) significant proliferation rate between fibroblast-like cells and SHED, (ii) high expression of fibroblast-associated markers in qRT-PCR analysis, and (iii) positive staining against collagen type 1, fibroblast-specific protein 1, and human thymic fibroblasts in flow cytometry analysis and immunofluorescence staining. The same expression patterns were found in primary human gingival fibroblasts, respectively. SHED as negative control showed lower expression or no signal, thus confirming the cells differentiated from SHED were fibroblast-like cells.
CONCLUSIONS: Taken together, the protocol adopted in this study suggests CTGF to be an appropriate inducer in the differentiation of SHED into fibroblast-like cells.
CLINICAL RELEVANCE: The fibroblast-like cells differentiated from SHED could be used in future in vitro and in vivo dental tissue regeneration studies as well as in clinical applications where these cells are needed.

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