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Combined effects of melatonin and FGF-2 on mouse preosteoblast behavior within interconnected porous hydroxyapatite ceramics - in vitro analysis.

Rahman MZ, Shigeishi H, Sasaki K, Ota A, Ohta K, Takechi M - J Appl Oral Sci (2016)

Bottom Line: The present in vitro study was undertaken to examine the relationship between these two bioactive agents and their combinatory effects on osteoblastic activity and mineralization in vitro.The synergy of the proliferative potential of FGF-2 and the differentiation potential of MEL showed increased osteogenic activity in MC3T3-E1 cells cultured within IP-CHA constructs.Conclusion These findings indicate that the combination of FGF-2 and MEL may be utilized with biocompatible materials to attain augmented osteogenic activity and mineralization.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Maxillofacial Surgery, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

ABSTRACT
Objective Biocompatible materials such as interconnected porous hydroxyapatite ceramics (IP-CHA) loaded with osteogenic cells and bioactive agents are part of an evolving concept for overcoming craniofacial defects by use of artificial bone tissue regeneration. Amongst the bioactive agents, melatonin (MEL) and basic fibroblast growth factor (FGF-2) have been independently reported to induce osteoblastic activity. The present in vitro study was undertaken to examine the relationship between these two bioactive agents and their combinatory effects on osteoblastic activity and mineralization in vitro. Material and Methods Mouse preosteoblast cells (MC3T3-E1) were seeded and cultured within cylindrical type of IP-CHA block (ø 4x7 mm) by vacuum-assisted method. The IP-CHA/MC3T3 composites were subjected to FGF-2 and/or MEL. The proliferation assay, alkaline phosphatase enzyme activity (ALP), mRNA expressions of late bone markers, namely Osteocalcin (OCN) and Osteopontin (OPN), and Alizarin Red staining were examined over a period of 7 days. Results FGF-2 mainly enhanced the proliferation of MC3T3-E1 cells within the IP-CHA constructs. MEL mainly induced the mRNA expression of late bone markers (OCN and OPN) and showed increased ALP activity of MC3T3 cells cultured within IP-CHA construct. Moreover, the combination of FGF-2 and MEL showed increased osteogenic activity within the IP-CHA construct in terms of cell proliferation, upregulated expressions of OCN and OPN, increased ALP activity and mineralization with Alizarin Red. The synergy of the proliferative potential of FGF-2 and the differentiation potential of MEL showed increased osteogenic activity in MC3T3-E1 cells cultured within IP-CHA constructs. Conclusion These findings indicate that the combination of FGF-2 and MEL may be utilized with biocompatible materials to attain augmented osteogenic activity and mineralization.

No MeSH data available.


Related in: MedlinePlus

Alizarin Red Staining of MC3T3-E1 cells under the influence of FGF-2 and/or melatonin (MEL). a- Alizarin Red staining of monolayer cultures showed increased mineralization after 2 weeks of treatment with MEL or FGF-2 in osteogenic induction medium compared to the control. The combination of MEL and FGF-2 resulted in more intense staining; b- Quantified values obtained from cells cultured within IP-CHA constructs suggested a role for MEL in mineralization, while combined FGF-2 and MEL induced increased mineralization. Statistical significance shown by p<0.05 is indicated by *
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f07: Alizarin Red Staining of MC3T3-E1 cells under the influence of FGF-2 and/or melatonin (MEL). a- Alizarin Red staining of monolayer cultures showed increased mineralization after 2 weeks of treatment with MEL or FGF-2 in osteogenic induction medium compared to the control. The combination of MEL and FGF-2 resulted in more intense staining; b- Quantified values obtained from cells cultured within IP-CHA constructs suggested a role for MEL in mineralization, while combined FGF-2 and MEL induced increased mineralization. Statistical significance shown by p<0.05 is indicated by *

Mentions: Figure 7a shows Alizarin Red Staining after MC3T3-E1 cells were treated for 2 weeks with MEL and/or FGF-2 in osteogenic induction medium. MEL and FGF-2 independently stimulated extracellular calcium deposition in both monolayer cultures and treated IP-CHA/MC3T3 composites (Figures 7a and 7b). Those in combination resulted in more intense mineralization staining and higher quantified values (Figures 7a and 7b). These results indicate that the combination of FGF-2 and MEL is involved in augmentation of mineralization.


Combined effects of melatonin and FGF-2 on mouse preosteoblast behavior within interconnected porous hydroxyapatite ceramics - in vitro analysis.

Rahman MZ, Shigeishi H, Sasaki K, Ota A, Ohta K, Takechi M - J Appl Oral Sci (2016)

Alizarin Red Staining of MC3T3-E1 cells under the influence of FGF-2 and/or melatonin (MEL). a- Alizarin Red staining of monolayer cultures showed increased mineralization after 2 weeks of treatment with MEL or FGF-2 in osteogenic induction medium compared to the control. The combination of MEL and FGF-2 resulted in more intense staining; b- Quantified values obtained from cells cultured within IP-CHA constructs suggested a role for MEL in mineralization, while combined FGF-2 and MEL induced increased mineralization. Statistical significance shown by p<0.05 is indicated by *
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4836923&req=5

f07: Alizarin Red Staining of MC3T3-E1 cells under the influence of FGF-2 and/or melatonin (MEL). a- Alizarin Red staining of monolayer cultures showed increased mineralization after 2 weeks of treatment with MEL or FGF-2 in osteogenic induction medium compared to the control. The combination of MEL and FGF-2 resulted in more intense staining; b- Quantified values obtained from cells cultured within IP-CHA constructs suggested a role for MEL in mineralization, while combined FGF-2 and MEL induced increased mineralization. Statistical significance shown by p<0.05 is indicated by *
Mentions: Figure 7a shows Alizarin Red Staining after MC3T3-E1 cells were treated for 2 weeks with MEL and/or FGF-2 in osteogenic induction medium. MEL and FGF-2 independently stimulated extracellular calcium deposition in both monolayer cultures and treated IP-CHA/MC3T3 composites (Figures 7a and 7b). Those in combination resulted in more intense mineralization staining and higher quantified values (Figures 7a and 7b). These results indicate that the combination of FGF-2 and MEL is involved in augmentation of mineralization.

Bottom Line: The present in vitro study was undertaken to examine the relationship between these two bioactive agents and their combinatory effects on osteoblastic activity and mineralization in vitro.The synergy of the proliferative potential of FGF-2 and the differentiation potential of MEL showed increased osteogenic activity in MC3T3-E1 cells cultured within IP-CHA constructs.Conclusion These findings indicate that the combination of FGF-2 and MEL may be utilized with biocompatible materials to attain augmented osteogenic activity and mineralization.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral and Maxillofacial Surgery, Institute of Biomedical and Health Sciences, Hiroshima University, Hiroshima, Japan.

ABSTRACT
Objective Biocompatible materials such as interconnected porous hydroxyapatite ceramics (IP-CHA) loaded with osteogenic cells and bioactive agents are part of an evolving concept for overcoming craniofacial defects by use of artificial bone tissue regeneration. Amongst the bioactive agents, melatonin (MEL) and basic fibroblast growth factor (FGF-2) have been independently reported to induce osteoblastic activity. The present in vitro study was undertaken to examine the relationship between these two bioactive agents and their combinatory effects on osteoblastic activity and mineralization in vitro. Material and Methods Mouse preosteoblast cells (MC3T3-E1) were seeded and cultured within cylindrical type of IP-CHA block (ø 4x7 mm) by vacuum-assisted method. The IP-CHA/MC3T3 composites were subjected to FGF-2 and/or MEL. The proliferation assay, alkaline phosphatase enzyme activity (ALP), mRNA expressions of late bone markers, namely Osteocalcin (OCN) and Osteopontin (OPN), and Alizarin Red staining were examined over a period of 7 days. Results FGF-2 mainly enhanced the proliferation of MC3T3-E1 cells within the IP-CHA constructs. MEL mainly induced the mRNA expression of late bone markers (OCN and OPN) and showed increased ALP activity of MC3T3 cells cultured within IP-CHA construct. Moreover, the combination of FGF-2 and MEL showed increased osteogenic activity within the IP-CHA construct in terms of cell proliferation, upregulated expressions of OCN and OPN, increased ALP activity and mineralization with Alizarin Red. The synergy of the proliferative potential of FGF-2 and the differentiation potential of MEL showed increased osteogenic activity in MC3T3-E1 cells cultured within IP-CHA constructs. Conclusion These findings indicate that the combination of FGF-2 and MEL may be utilized with biocompatible materials to attain augmented osteogenic activity and mineralization.

No MeSH data available.


Related in: MedlinePlus