Erken Osteojenik Farklılaşma (In Vitro) Süresince Wistar Sıçanlarda (Rattus novergicus) Gingival Medicinal Signaling Hücrelere Post Trombositten Zengin Fibrin Uygulamasının Agrekan Ekspresyonuna Etkisi
Trombositten Zengin Fibrin (TZF) büyüme faktörlerince zengin olup Gingival Medicinal Signaling Hücreleri (GMSH)’nin (GMSH) osteojenik farklılaşmasını geliştirebilir. Agrekan, kondrojenik farklılaşma markırı olup GMSH’nin osteojenik farklılaşmasının erken evrelerinde önemli bir rol oynamaktadır. Bu çalışma, TZF uygulaması sonrasında in vitro osteojenik farklılaşmada Agrekan ekspresyonunu araştırmayı amaçlamaktadır. Bu araştırma, post-test sadece kontrol grup dizayn kullanan rastgele örneklemeli gerçek bir deneysel çalışmadır. Yaklaşık 250 gram gelen bir aylık sağlıklı erkek Wistar sıçanların (Rattus novergicus) (n=4) alt gingival dokularından GMSH izole edildi, sonrasında 2 hafta kültüre edildi ve 4-5 gün pasajlandı. 3-5. pasajlarda GMSH, beş M24 besiyeri içerisinde 7, 14 ve 21 gün boyunca üç farklı kültür medyumunda (α Modified Eagle Medyum içeren negatif kontrol grubu, osteojenik medyum katkılı High Glucose-Dulbecco’s Modified Eagle Medyum (DMEM-HG) içeren pozitif kontrol grubu, hem osteojenik medyum hem de TZF katkılı DMEM-HG içeren uygulama grubu) kültüre edildi. Tek yönlü varyans analizi (ANOVA) testi uygulandı (P
The Aggrecan Expression Post Platelet Rich Fibrin Administration in Gingival Medicinal Signaling Cells in Wistar Rats (Rattus novergicus) During the Early Osteogenic Diffrentiation (In Vitro)
Platelet Rich Fibrin (PRF) is rich for growth factors which can improve the Gingival Medicinal Signaling Cells’ (GMSCs) osteogenic diffrentiation.Aggrecan is chondrogenic diffrentiation marker which has a significant role in the early stage of GMSCs’ osteogenic diffrentiation. This study aimed to analyze the expression of Aggrecan post PRF administration on the osteogenic diffrentiation in vitro. This research is a true experimental study using the post-test only control group design with a simple random sampling. GMSCs were isolated from the lower gingival tissue of healthy male Wistar rats (Rattus novergicus) (n=4), weighted around 250 g, a month old, then cultured for 2 weeks and passaged for 4-5 days. GMSCs in the passage 3-5 were cultured in five M24 plates (N=54; n=6/group) for 7 days, 14 days, and 21 days in three diffrent culture mediums, they were negative control group which included α Modified Eagle Medium; positive control group which contained High Glucose-Dulbecco’s Modified Eagle Medium (DMEMHG) combined with osteogenic medium; and at last, treatment group which were DMEM-HG combined with both osteogenic medium and PRF. A one-way Analysis of Variance (ANOVA) test (P
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- 1. Nugraha AP, Rantam FA, Ernawati DS, Narmada IB, Widodo ADW,
Lestari P, Dinaryanti A, Hendrianto E, Ihsan IS, Susilowati H, Karsari
D: Gingival mesenchymal stem cells from wistar rat’s gingiva (rattus
novergicus) isolation and characterization (in vitro study). J Int Dent Med
Res, 11 (2): 694-699, 2018.
- 2. Fitzsimmons REB, Mazurek MS, Soos A, and Simmons CA:
Mesenchymal stromal/stem cells in regenerative medicine and tissue
engineering. Stem Cells Int, 2018:8031718, 2018 DOI: 10.1155/2018/8031718
- 3. Caplan AI: Mesenchymal stem cells: Time to change the name! Stem
Cells Transl Med, 6 (6): 1445-1451, 2017. DOI: 10.1002/sctm.17-0051
- 4. Egusa H, Sonoyama W, Nishimura M, Atsuta I, Akiyama K: Stem cells
in dentisry - Part I: Stem cells sources, J Prosthodont Res, 56 (3): 151-165,
2012. DOI: 10.1016/j.jpor.2012.06.001
- 5. Nugraha AP, Narmada IB, Ernawati DS, Dinaryanti A, Hendrianto
E, Riawan W, Rantam FA: Bone alkaline phosphatase and osteocalcin
expression of rat’s gingival mesenchymal stem cells cultured in plateletrich fibrin for bone remodeling (in vitro study). Eur J Dent, 12 (4): 566-573,
2018. DOI: 10.4103/ejd.ejd_261_18
- 6. Nugraha AP, Narmada IB, Ernawati DS, Dinaryanti A, Hendrianto E,
Ihsan IS, Rantam FA: Osteogenic potential of gingival stromal progenitor
cells cultured in platelet rich fibrin is predicted by core-binding factor
subunit-α1/sox9 expression ratio (in vitro). F1000Res, 7:1134, 2018. DOI:
10.12688/f1000research.15423.1
- 7. Jing Y, Jing J, Ye L, Liu X, Harris SE, Hinton RJ, Feng JQ: Chondrogenesis and osteogenesis are one continuous developmental and
lineage defined biological process. Sci Rep, 7:10020, 2017. DOI: 10.1038/
s41598-017-10048-z
- 8. Qi H, Aguiar DJ, Williams SM, LA Pean A, Pan W, Verfaillie CM:
Identification of genes responsible for osteoblast differentiation from
human mesodermal progenitor cells. Proc Natl Acad Sci U S A, 100 (6):
3305-3310, 2003. DOI: 10.1073/pnas.0532693100
- 9. Namkoong B, Güven S, Ramesan S, Liaudanskaya V, Abzhanov A,
Demirci U: Recapitulating cranial osteogenesis with neural crest cells in
3-d microenvironments. Acta Biomater, 31, 301-311, 2016 DOI: 10.1016/j.
actbio.2015.12.004
- 10. Lauing KL, Cortes M, Domowicz MS, Henry JG, Baria AT, Schwartz NB:
Aggrecan is required for growth plate cytoarchitecture and diffrentiation.
Dev Biol, 396 (2): 224-236, 2014. DOI: 10.1016/j.ydbio.2014.10.005
- 11. Gibson BG, Briggs MD: The aggrecanopathies; an evolving phenotypic spectrum of human genetic skeletal diseases. Orphanet J Rare Dis,
11:86, 2016. DOI: 10.1186/s13023-016-0459-2
- 12. Sato T, Kudo T, Ikehara Y, Ogawa H, Hirano T, Kiyohara K, Hagiwara
K, Togayachi A, Ema M, Takahashi S, Kimata K, Watanabe H, Narimatsu
H: Chondroitin sulfate N-acetylgalactosaminyltransferase 1 is necessary
for normal endochondral ossification and aggrecan metabolism. J Biol
Chem, 286 (7): 5803-5812, 2011. DOI: 10.1074/jbc.M110.159244
- 13. Brunetti G, Mori G, D’Amelio P, Faccio R: The crosstalk between
the bone and the immune system: Osteoimmunology. Clin Dev Immunol,
2013:617319, 2013. DOI: 10.1155/2013/617319
- 14. Sumarta NPM, Pramono DC, Hendrianto E, Susilowati H, Karsari
D, Rantam FA: Chondrogenic diffrentiation capacity of human umbilical
cord mesenchymal stem cells with platelet rich fibrin scaffld in cartilage
regeneration (in vitro study). Bali Med J, 5 (3): 420-426, 2016. DOI:
10.15562/bmj.v5i3.295
- 15. Nugraha AP, Narmada IB, Ernawati DS, Dinaryanti A, Hendrianto
E, Ihsan IS, Riawan W, Rantam FA: In vitro bone sialoprotein-I expression
in combined gingival stromal cells and platelet rich fibrin during
osteogenic diffrentiation. Trop J Pharm Res, 17 (12): 2341-2345, 2018.
- 16. Sakata R, Iwakura T, Reddi AH: Regeneration of articular cartilage
surface: Morphogens, cells, and extracellular matrix scafflds. Tissue Eng
Part B Rev, 21 (5): 461-473, 2015. DOI: 10.1089/ten.TEB.2014.0661
- 17. Utomo DN, Mahyudin F, Hernugrahantoa KD, Suroto H, Chilmia
MZ, Rantam FA: Implantation of platelet rich fibrin and allogenic
mesenchymal stem cells facilitate the healing of muscle injury: An
experimental study on animal. Int J Surg, 11, 4-9, 2018. DOI: 10.1016/j.
ijso.2018.03.001