Potential differentiation of three types of primitive cells originated from different proliferation terms of mouse blood

The aim of this study was to differentiate central blood system mononucleated cells in vitro into osteoblast and osteoclast cells for three different proliferation terms of cells. The mononucleated cells were cultured in a selective proliferation medium for three different proliferation terms, i.e., short (5 days), medium (15 days) and long term (30 days) prior to analysis of osteoblast and osteoclast cells’ differentiation potentialities. The proliferation medium was then supplemented with differentiation factors, i.e., ascorbic acid and β-glycerophosphate to differentiate mononucleated cells into osteoblast cells. For osteoclast assay, RANKL and M-CSF were added into proliferation medium. For control, the same cells were used without supplementation of respective differentiation factors. The viability of differentiated cells from short, medium and long types of cells showed that they were able to survive until 10 to 14 days in the presence of respective differentiation factors without significant increased in the specific differentiation medium. Biochemical analyses on both alkaline phosphatase (ALP) and tartrate resistant acid phosphatase (TRAP) activities were significantly increased (p<0.05) once cultured in their respective differentiation medium. In conclusion, the three types of primitive cells have the same potentiality to differentiate into mature osteoblast and osteoclast cells even though the proliferation rates are different, i.e. 0.37, 0.55 and 0.72 division/day for short, medium and long term cells respectively. Mononucleated cells isolated from peripheral blood are primitive enough to differentiate into two distinct types of mature cells which originated from two different stem cells lineage hence can be categorized as multipotent stem cells

Stem Cell Heterogeneity of Mononucleated Cells from Murine Peripheral Blood: Molecular Analysis

The main purpose of this paper was to determine the heterogeneity of primary isolated mononucleated cells that originated from the peripheral blood system by observing molecular markers. The isolated cells were cultured in complete medium for 4 to 7 days prior to the separation of different cell types, that is, adherent and suspension. Following a total culture time of 14 days, adherent cells activated the Cd105 gene while suspension cells activated the Sca-1 gene. Both progenitor markers, Cbfa-1 and Ostf-1, were inactivated in both suspension and adherent cells after 14-day culture compared to cells cultured 3 days in designated differentiation medium. In conclusion, molecular analyses showed that primary mononucleated cells are heterogeneous, consisting of hematopoietic stem cells (suspension) and mesenchymal stem cells (adherent) while both cells contained no progenitor cells

Pengkulturan dan pencirian penanda molekul sel stem manusia daripada pulpa gigi susu (SHED) dan gigi kekal (DPSC)

Sel stem pulpa gigi manusia yang dipencilkan daripada tisu pulpa gigi merupakan sel stem dewasa bersifat multipoten. Objektif kajian ini adalah untuk mengenal pasti teknik pengkulturan in vitro sel stem pulpa gigi susu (SHED) dan gigi kekal (DPSC) melalui penentuan pasaj, kesan eraman tripsin-EDTA dan potensi proliferasi serta untuk mencirikan kedua-dua sel ini melalui profil penanda molekul. Kaedah pencernaan enzim digunakan pada tisu pulpa gigi susu dan gigi kekal masing-masing untuk pemencilan sel SHED dan DPSC. Kedua-dua sel dikulturkan daripada pasaj 1 hingga 5 dan pewarnaan tripan biru digunakan untuk memperoleh lengkuk pertumbuhan bagi menentukan masa penggandaan populasi sel (PDT) pada setiap pasaj. Kesan tripsin-EDTA terhadap kedua-dua sel dikaji menggunakan pewarnaan Alamar biru untuk menentukan masa eraman yang optimum semasa proses pengsubkulturan. Potensi proliferasi in vitro bagi kedua-dua sel selama 21 hari ditentukan melalui asai 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolium bromida (MTT). Pencirian kedua-dua sel melalui pengekspresan penanda biologi molekul ditentukan melalui pendekatan RT-PCR. Morfologi kedua-dua sel didapati menyerupai sel fibroblas pada kesemua pasaj. Sel SHED dan DPSC pada pasaj 3 menunjukkan PDT terendah iaitu masing-masing 43 ± 2.3 dan 63 ± 3.1 jam. Pendedahan SHED terhadap tripsin-EDTA menunjukkan penurunan peratus sel viabel berbanding DPSC. Pertumbuhan sel SHED didapati ~2.3 kali ganda lebih tinggi berbanding DPSC. Pencirian molekul kedua-dua sel menunjukkan pengekspresan penanda sel stem mesekima dan bukannya penanda sel stem hematopoietik. Kesimpulannya, morfologi sel yang homogenus dan nilai PDT terendah yang ditunjukkan oleh sel daripada pasaj 3 menjadikannya pasaj terbaik untuk menentukan potensi proliferasi sel dan pengekpresan penanda molekul. SHED didapati mampu berproliferasi dengan lebih baik berbanding DPSC. Walau bagaimanapun, DPSC lebih rentan terhadap tripsin-EDTA berbanding SHED. Pencirian molekul pula mendapati kedua-dua sel merupakan sel stem jenis mesenkima

Cellular and Molecular Changes in Orthodontic Tooth Movement

Tooth movement induced by orthodontic treatment can cause sequential reactions involving the periodontal tissue and alveolar bone, resulting in the release of numerous substances from the dental tissues and surrounding structures. To better understand the biological processes involved in orthodontic treatment, improve treatment, and reduce adverse side effects, several of these substances have been proposed as biomarkers. Potential biological markers can be collected from different tissue samples, and suitable sampling is important to accurately reflect biological processes. This paper covers the tissue changes that are involved during orthodontic tooth movement such as at compression region (involving osteoblasts), tension region (involving osteoclasts), dental root, and pulp tissues. Besides, the involvement of stem cells and their development towards osteoblasts and osteoclasts during orthodontic treatment have also been explained. Several possible biomarkers representing these biological changes during specific phenomenon, that is, bone remodelling (formation and resorption), inflammation, and root resorption have also been proposed. The knowledge of these biomarkers could be used in accelerating orthodontic treatment

Intrinsic anticarcinogenic effects of Piper sarmentosum ethanolic extract on a human hepatoma cell line

<p>Abstract</p> <p>Background</p> <p><it>Piper sarmentosum</it>, locally known as kaduk is belonging to the family of Piperaceae. It is our interest to evaluate their effect on human hepatoma cell line (HepG2) for the potential of anticarcinogenic activity.</p> <p>Results</p> <p>The anticarcinogenic activity of an ethanolic extract from <it>Piper sarmentosum </it>in HepG2 and non-malignant Chang's liver cell lines has been previously determined using (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide) (MTT) assays, where the IC₅₀value was used as a parameter for cytotoxicity. The ethanolic extract that showed anticarcinogenic properties in HepG2 cells had an IC₅₀of 12.5 μg mL^-1, while IC₅₀values in the non-malignant Chang's liver cell line were greater than 30 μg mL^-1. Apoptotic morphological changes in HepG2 cells were observed using an inverted microscope and showed chromatin condensation, cell shrinkage and apoptotic bodies following May-Grunwald-Giemsa's staining. The percentage of apoptotic cells in the overall population (apoptotic index) showed a continuously significant increase (p < 0.05) in 12.5 μg mL^-1ethanolic extract-treated cells at 24, 48 and 72 hours compared to controls (untreated cells). Following acridine orange and ethidium bromide staining, treatment with 10, 12 and 14 μg mL^-1of ethanolic extracts caused typical apoptotic morphological changes in HepG2 cells. Molecular analysis of DNA fragmentation was used to examine intrinsic apoptosis induced by the ethanolic extracts. These results showed a typical intrinsic apoptotic characterisation, which included fragmentation of nuclear DNA in ethanolic extract-treated HepG2 cells. However, the non-malignant Chang's liver cell line produced no DNA fragmentation. In addition, the DNA genome was similarly intact for both the untreated non-malignant Chang's liver and HepG2 cell lines.</p> <p>Conclusion</p> <p>Therefore, our results suggest that the ethanolic extract from <it>P. sarmentosum </it>induced anticarcinogenic activity through an intrinsic apoptosis pathway in HepG2 cells <it>in vitro</it>.</p

Differentiation analyses of adult suspension mononucleated peripheral blood cells of Mus musculus

<p>Abstract</p> <p>Background</p> <p>The purpose of this study is to determine whether isolated suspension mouse peripheral mononucleated blood cells have the potential to differentiate into two distinct types of cells, i.e., osteoblasts and osteoclasts.</p> <p>Results</p> <p>Differentiation into osteoblast cells was concomitant with the activation of the <it>Opn </it>gene, increment of alkaline phosphatase (ALP) activity and the existence of bone nodules, whereas osteoclast cells activated the <it>Catk </it>gene, increment of tartrate resistant acid phosphatase (TRAP) activity and showed resorption activities via resorption pits. Morphology analyses showed the morphology of osteoblast and osteoclast cells after von Kossa and May-Grunwald-Giemsa staining respectively.</p> <p>Conclusions</p> <p>In conclusion, suspension mononucleated cells have the potentiality to differentiate into mature osteoblasts and osteoclasts, and hence can be categorized as multipotent stem cells.</p

In Vitro Chondrogenesis Transformation Study of Mouse Dental Pulp Stem Cells

A major challenge in the application of mesenchymal stem cells in cartilage reconstruction is that whether the cells are able to differentiate into fully mature chondrocytes before grafting. The aim of this study was to isolate mouse dental pulp stem cells (DPSC) and differentiate them into chondrocytes. For this investigation, morphological, molecular, and biochemical analyses for differentiated cells were used. To induce the chondrocyte differentiation, DPSC were cultured in chondrogenic medium (ZenBio, Inc.). Based on morphological analyses using toluidine blue staining, proteoglycan products appear in DPSC after 21 days of chondrocyte induction. Biochemical analyses in differentiated group showed that alkaline phosphatase activity was significantly increased at day 14 as compared to control (P &lt; 0.05). Cell viability analyses during the differentiation to chondrocytes also showed that these cells were viable during differentiation. However, after the 14th day of differentiation, there was a significant decrease (P &lt; 0.05) in the viability proportion among differentiated cells as compared to the control cells. In RT-PCR molecular analyses, mouse DPSC expressed Cd146 and Cd166 which indicated that these cells belong to mesenchymal stem cells. Coll I and Coll II markers showed high expression after 14 and 21 days, respectively. In conclusion, this study showed that DPSC successfully differentiated into chondrocytes

In Vitro

A major challenge in the application of mesenchymal stem cells in cartilage reconstruction is that whether the cells are able to differentiate into fully mature chondrocytes before grafting. The aim of this study was to isolate mouse dental pulp stem cells (DPSC) and differentiate them into chondrocytes. For this investigation, morphological, molecular, and biochemical analyses for differentiated cells were used. To induce the chondrocyte differentiation, DPSC were cultured in chondrogenic medium (Zen-Bio, Inc.). Based on morphological analyses using toluidine blue staining, proteoglycan products appear in DPSC after 21 days of chondrocyte induction. Biochemical analyses in differentiated group showed that alkaline phosphatase activity was significantly increased at day 14 as compared to control (P<0.05). Cell viability analyses during the differentiation to chondrocytes also showed that these cells were viable during differentiation. However, after the 14th day of differentiation, there was a significant decrease (P<0.05) in the viability proportion among differentiated cells as compared to the control cells. In RT-PCR molecular analyses, mouse DPSC expressed Cd146 and Cd166 which indicated that these cells belong to mesenchymal stem cells. Coll I and Coll II markers showed high expression after 14 and 21 days, respectively. In conclusion, this study showed that DPSC successfully differentiated into chondrocytes

Kesan penyimpanan sampel air liur terhadap kualiti DNA genom

Air liur berpotensi menjadi punca DNA yang mudah diambil bagi kajian klinikal kerana tidak invasif berbanding sampel darah. Kajian ini dijalankan untuk memencilkan dan menulenkan DNA genom daripada sampel air liur manusia serta mengkaji kesan penyimpanan terhadap kualiti DNA genom. Sampel air liur (n=5) disimpan dalam penimbal Tris-NaCl EDTA (TNE) pada suhu bilik (25°C) mengikut tempoh masa yang ditetapkan iaitu, segar (tanpa penyimpanan), 1, 2, 3 dan 4 bulan. Pemencilan dan penulenan DNA dilakukan menggunakan kaedah fenol-kloroform. Seterusnya, PCR telah dijalankan untuk mengetahui ketulenan DNA yang diekstrak menggunakan amplifikasi pada kawasan jujukan beta-globin dan mengenal pasti kehadiran bakteria melalui jujukan yang mengekod 16S rDNA. Keputusan menunjukkan fragmen DNA gen beta-globin manusia hanya berjaya diamplifikasi daripada sampel segar. Sampel air liur yang disimpan dalam penimbal TNE pada suhu bilik tidak mampu menstabilkan DNA genom manusia untuk jangka masa lama dan hanya berkesan untuk tempoh yang singkat iaitu, kurang daripada 1 bulan. Kesimpulannya, hanya sampel air liur segar sahaja yang berupaya memencil DNA genom

Scaffold selection for tissue engineering in dentistry

Tissue engineering aims to restore lost, damaged, diseased or defective tissues in the human body using engineered or regenerated products. The advancement of tissue engineering has given a promising opportunity for better clinical practice in treating dental patients especially in the fields of endodontic, bone and periodontal tissue as well as whole tooth regeneration. In this review, we briefly summarise the possible selection criteria of scaffolds for potential tissue engineering applications in dentistry. Biochemical and physical properties, as well as scaffolding approaches involved in the selection of an ideal scaffold for dental tissue engineering, are also discussed in this review. This review also discussed major applications of tissue engineering in the dentistry field, which can create a paradigm for future studies of tissue regeneration by using selected cells and scaffolds as an alternative treatment in dentistry