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Fulltext Collagen type I promotes osteogenic differentiation of amniotic membrane-derived mesenchymal stromal cells in basal and induction media

Akhir HM, Teoh PL

Biosci Rep, 2020 12 23;40(12).
PMID: 33245097 DOI: 10.1042/BSR20201325

Collagen has been widely shown to promote osteogenesis of bone marrow mesenchymal stromal cells (BM-MSCs). Due to the invasive procedure of obtaining BM-MSCs, MSCs from other tissues have emerged as a promising alternative for regenerative therapy. MSCs originated from different sources, exhibiting different differentiation potentials. Therefore, the applicability of collagen type I (COL), combining with amniotic membrane (AM)-MSCs was examined through proliferation and differentiation assays together with the expression of surface markers and genes associated with stemness and differentiation under basal or induction conditions. No increase in cell growth was observed because AM-MSCs might be directed toward spontaneous osteogenesis. This was evidenced by the calcium deposition and elevated expression of osteogenic genes when AM-MSCs were cultured in collagen plate with basal media. Under the osteogenic condition, reciprocal expression of OCN and CEBPA suggested a shift toward adipogenesis. Surprisingly, adipogenic genes were not elevated upon adipogenic induction, although oil droplets deposition was observed. In conclusion, our findings demonstrated that collagen causes spontaneous osteogenesis in AM-MSCs. However, the presence of exogenous inductors could shift the direction of adipo-osteogenic gene regulatory network modulated by collagen.
Fulltext WDR5, ASH2L, and RBBP5 control the efficiency of FOS transcript processing

Teoh PL, Sharrocks AD

Cell Mol Biol Lett, 2014 Jun;19(2):215-32.
PMID: 24715476 DOI: 10.2478/s11658-014-0190-8

H3K4 trimethylation is strongly associated with active transcription. The deposition of this mark is catalyzed by SET-domain methyltransferases, which consist of a subcomplex containing WDR5, ASH2L, and RBBP5 (the WAR subcomplex); a catalytic SET-domain protein; and additional complexspecific subunits. The ERK MAPK pathway also plays an important role in gene regulation via phosphorylation of transcription factors, co-regulators, or histone modifier complexes. However, the potential interactions between these two pathways remain largely unexplored. We investigated their potential interplay in terms of the regulation of the immediate early gene (IEG) regulatory network. We found that depletion of components of the WAR subcomplex led to increased levels of unspliced transcripts of IEGs that did not necessarily reflect changes in their mature transcripts. This occurs in a manner independent from changes in the H3K4me3 levels at the promoter region. We focused on FOS and found that the depletion of WAR subcomplex components affected the efficiency of FOS transcript processing. Our findings show a new aspect of WAR subcomplex function in coordinating active transcription with efficient pre-mRNA processing.
Differential gene expression of Wharton's jelly-derived mesenchymal cells mediated by graphene oxide in basal and osteo-induced media

Hiew VV, Teoh PL

Mol Biol Rep, 2024 Mar 03;51(1):383.
PMID: 38433142 DOI: 10.1007/s11033-024-09324-9

BACKGROUND: Graphene oxide (GO) is widespread in scaffold engineering owing to its extraordinary properties such as multiple oxygen functional groups, high hydrophilicity ability and biocompatibility. It is known to promote differentiation in mesenchymal stem cells, but concomitant comparison of its modulation on the expression profiles of Wharton's jelly (WJ)-MSC surface markers, lineage differentiation, and epigenetic regulatory genes in basal and induced condition are still lacking. Unraveling the fundamental mechanisms is essential for the effective utilization of WJ-MSCs incorporated with GO in therapy. This study aims to explore the unique gene expression profiles and epigenetic characteristics of WJ-MSCs influenced by GO.
METHODS AND RESULTS: The characterized GO-coated coverslip served as a substrate for culturing WJ-MSCs. In addition to investigating the impact of GO on cell proliferation and differentiation, we conducted a gene expression study using PCR array, while epigenetic control was assessed through bisulfite sequencing and Western blot analysis. Our findings indicate that the presence of GO maintained the proliferation and survival of WJ-MSCs. In the absence of induction, GO led to minor lipid and glycosaminoglycan deposition in WJ-MSCs. This was evidenced by the sustained expression of pluripotency and lineage-specific genes, demethylation at the OCT4 promoter, and a decrease in H3K9 methylation. In osteo-induced condition, the occurrence of osteogenesis appeared to be guided by BMP/TGF and ERK pathway activation, accompanied by the upregulation of osteogenic-related genes and downregulation of DNMT3b.
CONCLUSIONS: GO in osteo-induced condition create a favorable microenvironment that promotes the osteogenesis of WJ-MSCs by influencing genetic and epigenetic controls. This helps in advancing our knowledge on the use of GO as priming platform and WJ-MSCs an alternate source for bone repair and regeneration.
Phyla nodiflora L. Extracts Induce Apoptosis and Cell Cycle Arrest in Human Breast Cancer Cell Line, MCF-7

Teoh PL, Liau M, Cheong BE

Nutr Cancer, 2019;71(4):668-675.
PMID: 30663402 DOI: 10.1080/01635581.2018.1559942

Phyla nodiflora L. has been used as medicinal remedies for various ailments due to its antioxidant, anti-inflammatory, anti-bacterial, anti-tumor activity. Previously, we found that the plant extracts induced DNA fragmentation in MCF-7. This study was to investigate the modes of action of P. nodiflora in inhibiting breast cancer cells using leaf ethyl acetate (EA leaf), stem ethyl acetate (EA stem) and stem methanol (Met stem) extracts. The MTT assay showed that the anti-proliferative effects of P. nodiflora extracts were selective towards MCF-7 with a minimal effect on MCF10A. Morphological changes such as cell shrinkage and nuclear condensation were observed in treated cells. We found that induction of apoptosis by EA leaf and EA stem was mitochondrial-dependent while loss of mitochondrial membrane potential was not found in Met stem-treated cells. In addition, the expression levels of AIFM1, CASP9, CFLAR, and IGF1R were altered after treatment. Decreased BCL-2 expression was found in treated cells while BAX and caspases' expression was upregulated or maintained. All extracts caused perturbation of cell cycle at S phase by dysregulating the expression of cell cycle regulators such as CDKs and cyclins. Our findings indicate that P. nodiflora inhibits MCF-7 cells by inducing apoptosis and perturbing cell cycle.
Stem Hexane Extract of Strobilanthes crispus Induces Apoptosis in Triple-Negative Breast Cancer Cell Line

Gordani N, Cheong BE, Teoh PL

Nutr Cancer, 2022;74(1):299-305.
PMID: 33511882 DOI: 10.1080/01635581.2021.1880606

Strobilanthes crispus is known to possess multiple health beneficial effects and reported to be traditionally used as medicine in several countries. This study was to investigate the anti-proliferative effects of S. crispus leaves and stem extracts on MDA-MB-231 by examining their effects on apoptosis pathway. The chemical compounds were extracted from leaves and stems using methanol followed by solvent partitioning. Two extracts were found to prevent MDA-MB-231 cell growth at the IC50 of 45 μg/mL and 60 μg/mL, respectively, for leaf water (LW) and stem hexane (SH) extracts. Results showed that SH extract induces apoptosis by suppressing the protein expression of BCL-2 while the expression of pro-apoptotic proteins such as BAX and caspase nine were unchanged. Decrease of cyclin A2 in SH-treated cells suggested this effect was associated with the dysregulation of cell cycle. However, LW extract showed no effects on apoptosis and cell cycle arrest in the treated cells. Taken together, our results showed SH extract of S. crispus exhibiting their anti-proliferative activities by modulating apoptosis and cell cycle, but the underlying mechanisms exerted by LW extract requires further investigation.
The Advancement of Biomaterials in Regulating Stem Cell Fate

Hiew VV, Simat SFB, Teoh PL

Stem Cell Rev Rep, 2018 Feb;14(1):43-57.
PMID: 28884292 DOI: 10.1007/s12015-017-9764-y

Stem cells are well-known to have prominent roles in tissue engineering applications. Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) can differentiate into every cell type in the body while adult stem cells such as mesenchymal stem cells (MSCs) can be isolated from various sources. Nevertheless, an utmost limitation in harnessing stem cells for tissue engineering is the supply of cells. The advances in biomaterial technology allows the establishment of ex vivo expansion systems to overcome this bottleneck. The progress of various scaffold fabrication could direct stem cell fate decisions including cell proliferation and differentiation into specific lineages in vitro. Stem cell biology and biomaterial technology promote synergistic effect on stem cell-based regenerative therapies. Therefore, understanding the interaction of stem cell and biomaterials would allow the designation of new biomaterials for future clinical therapeutic applications for tissue regeneration. This review focuses mainly on the advances of natural and synthetic biomaterials in regulating stem cell fate decisions. We have also briefly discussed how biological and biophysical properties of biomaterials including wettability, chemical functionality, biodegradability and stiffness play their roles.
Chemical composition and cytotoxic properties of Clinacanthus nutans root extracts

Teoh PL, Cheng AY, Liau M, Lem FF, Kaling GP, Chua FN, et al.

Pharm Biol, 2017 Dec;55(1):394-401.
PMID: 27931178

CONTEXT: Clinacanthus nutans Lindau (Acanthaceae) is a medicinal plant that has been reported to have anti-inflammatory, antiviral, antimicrobial and antivenom activities. In Malaysia, it has been widely claimed to be effective in various cancer treatments but scientific evidence is lacking.
OBJECTIVE: This study investigates the chemical constituents, anti-proliferative, and apoptotic properties of C. nutans root extracts.
MATERIALS AND METHODS: The roots were subjected to solvent extraction using methanol and ethyl acetate. The anti-proliferative effects of root extracts were tested at the concentrations of 10 to 50 μg/mL on MCF-7 and HeLa by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay for 72 h. Morphological changes were observed under light microscope. Pro-apoptotic effects of root extracts were examined using flow cytometric analysis and RT-PCR. The chemical compositions of root extracts were detected using GC-MS.
RESULTS: The proliferation of MCF-7 cells was inhibited with the IC50 values of 35 and 30 μg/mL, respectively, for methanol and ethyl acetate root extracts. The average inhibition of HeLa cells was ∼25%. Induction of apoptosis in MCF-7 was supported by chromatin condensation, down-regulation of BCL2 and unaltered expression of BAX. However, only ethyl acetate extract caused the loss of mitochondrial membrane potential. GC-MS analysis revealed the roots extracts were rich with terpenoids and phytosterols.
DISCUSSION AND CONCLUSIONS: The results demonstrated that root extracts promote apoptosis by suppressing BCL2 via mitochondria-dependent or independent manner. The identified compounds might work solely or cooperatively in regulating apoptosis. However, further studies are required to address this.
Community pharmacists' knowledge, attitudes and practices towards oral healthcare and its management: A systematic review

Rajiah K, Lim WK, Madeline Teoh PL, Binti Mas'od MA, Lim WY, Poh Chou LL, et al.

Int J Clin Pract, 2021 Sep;75(9):e14096.
PMID: 33619786 DOI: 10.1111/ijcp.14096

BACKGROUND: Community pharmacists play a significant part in providing oral healthcare advice as there are significant connections between oral healthcare and body system healthcare.
OBJECTIVE: To systematically review the literature regarding the knowledge, attitudes and practice of community pharmacists in managing oral healthcare problems.
METHODS: A systematic review was conducted through Scopus, PubMed and Google Scholar databases. Studies regarding knowledge, attitudes and practice of management of dental care by community pharmacists between 1990 and 2019 were included.
RESULTS: Forest plot was performed to access knowledge, attitudes and practice of community pharmacist on oral healthcare. The results showed there were 44% of community pharmacists have a lack of knowledge on oral healthcare to provide an appropriate recommendation to patients with dental problems. Eighty-eight per cent of community pharmacists were willing to improve their knowledge of oral healthcare. There were 86% of them recognised that their role was significant in oral health. However, there were 59% of community pharmacists who had poor attitude in providing oral health information.
CONCLUSIONS: Community pharmacists were lacking knowledge on oral health mainly because of paucity in providing appropriate training courses. This has led to poor practices towards oral healthcare as they were unable to provide suitable products recommendations to the patients. This has led the community pharmacists into lack of attitudes in providing oral health information. However, most of the community pharmacists were conscious of their role in the oral healthcare system and were willing to improve their knowledge of oral healthcare.