Lecture Note
The Effect of Magnesium on Dentinogesis of Human Dentalpulp Cells (Materials and Methods)
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University:
Boston University -
Course:
GMS BC 630 | Ethical & Policy Issues in Health and Medical Services Emergency Management Academic year:
2020
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Views:
419
Pages:
18
Author:
Clint W.
MATERIALS & METHODS Preparation of Magnesium Chloride
Magnesium
Chloride
Magnesium
Chloride Steps of Human Dental Pulp Cells (HDPCS)
Extraction
G
H Human Dental Pulp Cells Cultured with Magnesium Chloride Concentrations
Freshly Extracted Third Molar Teeth
Primary Passage of Human Dental Pulp Cell Culture
Secondary Passage of Human Dental Pulp Cell Culture
Magnesium Chloride
(MgCl2)
Control
Control
0.5 mM
MgCl2
1 mM
MgCl2
2 mM
MgCl2
4 mM
MgCl2
8 mM
MgCl2
Outcome measures
Attachment Efficiency
4, 8,12,16 and 20 hrs
Proliferation
7,10,14 days
Differentiation
7,10,14 days
ALP DSP DMP1 DSPP
Mineralization
7,10,14 days RESULTS
RESULTS 120
Cell Attachment Effeciency (%)
100
80
60
40
20
0
Control
0.5 mm
1mm
2mm
4mm
8mm
Concentrations of Supplemental Magnesium Chloride (MgCl2) mM
Histogram showing Cell Attachment Efficiency at 16 hours of all Magnesium Chloride (MgCl2) Concentrations Control
0.5 mM
1 mM
2 mM
4 mM
8 mM
50
b
b
45
b
40
b
35
b
Cell Proliferation (Folds).
30
b
b
b
c
c
25
20
c
c
15
a
10
5
0
a
a
a
a
a
Day 3
b
b
a
a
d
Day 7
c
d
Day 10
d
d
d
Day 14
Histogram showing Cell Proliferation Rate of Magnesium Chloride (MgCl2) at Different Time Intervals
d
d
Day 21
Time (Days) Control
0.5 mM
1 mM
2 mM
4 mM
8 mM
120
a
a
100
b
b
Cell Viability %
80
60
a
a
b c
c
a
b
b
b
b
b
b
c
c
d
d
e
40
f
e
f
e
f
c
d
d
20
0
16 hrs
Day 3
Day 7
Day 10
Day 14
Time (Days)
Histogram showing Cell Viability of Magnesium Chloride (MgCl2) at Different Time Intervals
d 1.4
Control
0.5 mM
1 mM
2 mM
4 mM
b
8 mM
1.2
b
Cell Proliferation (Folds)
1
0.8
b
0.6
b
0.4
b
a
b
a
a
a
c
a
a
d
a
a
e
d
0.2
0
Day 7
Day 10
Day 14
P<0.0001
Time (Days)
Histogram showing Cell Proliferation Rate of Magnesium Chloride (MgCl2) at Different Time Intervals
(Odontogenic Media) Control
3.5
0.5 mM
1 mM
2 mM
4 mM
8 mM
b
3
c
ALP Activity (mU/ml ) Per Million Cells
2.5
b
b
c
c
b
a
d
2
d
1.5
c
d
1
0.5
0
a
b
a
a
e
f
Day 7
Day 10
Day 14
P<0.0001
Time (Days)
Histogram showing Alkaline Phosphatase Activity of Magnesium Chloride (MgCl2) at Different Time
Intervals Dentin Sialoprotein Expression (ng/ml) Per Million Cells
Control
0.5 mM
1 mM
2 mM
4 mM
8 mM
500
b
450
c
d
400
350
300
b
250
c
d
a
e
e
f
f
200
150
100
50
0
c
a
b
Day 7
d
a
a
e
Day 10
Day 14
Time (Days)
Histogram showing Dentin Sialoprotein (DSP) Expression of Magnesium Chloride (MgCl2) at Different Time
Intervals Control
600000
0.5 mM
1 mM
2 mM
4 mM
8 mM
c
b
500000
Dentin Matrix Protein Expression (DMP-1)
(pg/ml) Per Million Cells
400000
b
c
a
a
b
c
d
100000
0
Day 7
e
e
a
d
f
e
300000
200000
d
f
f
Day 10
Day 14
P<0.0001
Time (Days)
Histogram showing Dentin matrix protein1(DMP-1) Expression of Magnesium Chloride (MgCl2)
at Different Time Intervals Dentin Sialo phospho (DSPP) Protein Expression (ng/ml) Per Million Cells
Control
0.5 mM
1 mM
2 mM
4 mM
8mM
500000000
c
450000000
b
400000000
d
350000000
300000000
c
250000000
b
200000000
50000000
0
a
b
d
e
a
150000000
100000000
a
c
d
e
e
Day 7
f
f
Day 10
Histogram showing Dentin Sialophosphoprotein (DSPP) Expression of Magnesium Chloride (MgCl2)
at Different Time Intervals
Day 14
P<0.0001
Time (Days)
f Minerlization Rate per million cells
Control
0.5 mM
1 mM
2 mM
4 mM
c
9
8
b
7
6
c
5
4
3
2
8 mM
a
b
c
c
c
d
b
d
d
e
d
e
a
f
a
1
0
Day 7
Day 10
Day 14
Time (Days)
Histogram showing Mineralization Rate of Magnesium Chloride (MgCl2) at Different Time Intervals CONCLUSIONS •
Conclusions
• This is the first report to investigate the optimal magnesium
concentration needed to induce human dental pulp cell
attachment, proliferation, differentiation and mineralization •
Conclusions
• This study showed that HDPCs with 0.5mM and 1 mM magnesium
supplemental groups elicited the highest stimulatory effect on cell attachment
efficiency, cell viability, proliferation rate, ALP activity, expression of
odontogenic-related proteins (DSP, DMP-1, DSPP) and mineralization at all
time points Conclusions
• However, 8mM magnesium group had an inhibitory effect on HDPCS
behavior showing lower ALP activity, expression of DSP, DMP-1, DSPP, and
mineralization compared to the control
The Effect of Magnesium on Dentinogesis of Human Dentalpulp Cells (Materials and Methods)
of 18
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