Osteoclast and Osteoblast Quantity Change in Guinea Pig’s Tooth

Movement

Erliera

, Aditya Rachmawati

, Veranyca Chiuman

Orthodonti Department, Faculty of Dentistry, Universitas Sumatera Utara,

Alumni Street No. 2 Universitas Sumatera Utara, Medan, Indonesia

Keywords: Remodelling, Osteoclast, Osteoblast.

Abstract: An orthodontic treatment is defined by the quantity and the quality of the remodelling process including bone

resorption by osteoclast, and deposition by osteoblast. The activities of osteoclast and osteoblast in

orthodontic tooth movement can be seen by quantitative analysis done to the alveolar bone histologic tissue

on guinea pig. The aim of this research is to observe when the cells’ activities in orthodontic tooth movement

reach its optimum to move the tooth. This is an experimental research which includes intervension and control

groups. Guinea pigs were divided into five groups by the time they be observed and were given new separator

on each session to produce continuous force. This research showed that osteoclast activities reach its

maximum at day 7, whereas the osteoblast activities is at day 21. The amount of osteoclast cells in five

timesperiod is not statistically different (p=0,061), while the amount of osteoblast cell is statistically different

(p=0,006). The difference of osteoclast cells’ amount in control groups is statistically different with the

intervention group at day 28 (p=0,028), whereas the osteoblast is statistically different at day 7 (p=0,015). In

conclusion, osteoclast cells reach its maximum amount at day 7, whereas the osteoblast at day 21. The amount

of osteoclast cells was highly increased when the intervention begins and decreased significantly through the

end of the research (day 28), whereas the amount of osteoblast cell in control group is decreased drastically

when the force was being applied (day 7), and had started to increase after day 14.

1 INTRODUCTION

Orthodontic treatment is one of the treatments which

is done in dentistry to achieve an aesthetic dentofacial

appearance by correcting the inter-incisal alignment,

producing a good occlusion, removing the

arrangement of crowded and rotated teeth (Ardhana,

2013). The movement of teeth during orthodontic

treatment causes an intercourse reaction between

periodontal tissue, alveolar bone, and causes the

release of certain substances from within the teeth and

the systems around it (Ariffin, 2011). Histologically,

the periodontal tissue receives forces which are

generally known in orthodontic as pressure area,

which is the area where the resorption of alveolar

bone happens due to the pressure on periodontal

ligament, and the tension area, which is the area

where the formation of the alveolar bone takes place.

Both of these forces are the ones that make the

movement of teeth possible (Harry and Sandy, 2004).

The basic orthodontic treatment can be done by

repositioning the teeth by either using fixed or

removable appliances (Hikmah, 2015). There are few

concepts describing the mechanism of orthodontic

tooth movement: pressure tension theory, blood flow

theory, bone bending and piezoelectric theory

(Bhalajhi, 2004). When an orthodontic force is

applied, the space of periodontal ligament in the

pressure area becomes smaller which causes vascular

constriction, reduction of cells replication and

collagen production, followed by resorption of the

bone (Meikle, 2006). Whereas in the tension area, the

space between periodontal ligament becomes wider,

vascularization increases, replication of cell and

production of collagen increases, thus causes

deposition of bone (Graber and Vanarsdall, 2012).

Orthodontic tooth movement are divided into

three phases: initial phase, lag phase, and post lag

phase (Krishnan and Davidovitch, 2006). Initial

phase is the phase which involves rapid movement of

the teeth. This phase causes teeth to move inside the

space of periodontal ligament and bending of alveolar

bone (Ariffin, 2011). In lag phase, there are no

movement of the teeth. But if it happens, it only

Erliera, ., Rachmawati, A. and Chiuman, V.

Osteoclast and Osteoblast Quantity Change in Guinea Pig’s Tooth Movement.

DOI: 10.5220/0010076405050508

In Proceedings of the International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches (ICOSTEERR 2018) - Research in Industry 4.0, pages

505-508

ISBN: 978-989-758-449-7

505

occurs in a short distance (Krishnan and Davidovitch,

2006). This movement is caused by the hyalinization

tissue process of the periodontal ligament that receive

force. The movement of teeth will not happen until

the cell completely resorb the whole necrotic tissue

area (Krishnan and Davidovitch, 2015). In post lag

phase, the speed of tooth movement increases

gradually. Tooth movement will reoccur after all the

hyaline area is eliminated and then the resorption will

then begin (Bhalajhi, 2004).

A successful orthodontic treatment is based on the

remodelling process which involves the process of

bone resorption by osteoclast, and bone deposition by

osteoblast (Proffit, 2007). These processes are based

on the quantity and quality of osteoclast and

osteoblast (Hikmah, 2016). Boulpaep and boron (cit

Kini and Nandeesh) stated that after putting the force

on ligament periodontal for a few hours, osteoclast

will be multiplied and resorption process will begin

on the bone surface, whereas for the involving tension

area, osteoblast will replicate and forms a new layer

of bone. Based on this situation, a successful

orthodontic treatment is based on the effectiveness of

remodelling process, which is based on the quality

and quantity of the osteoclast and osteoblast (Kini and

Nandeesh, 2012).

Activation of osteoblast and osteoclast cells can

be observed in a few ways: histology examination

through phatology anatomy with biomarker protein

coloring, or with the coloring of Hematoxylin Eosin

(HE). Samples which are going to be examined is

firstly fixated using formalin, then going through

decalcification, dehydration, clearing, and

embedding processes. Samples are then cut

longitudinally by using microtom to identify the

histologic image of osteoblast and osteoclast cell.

Generally, cells that are analyzed histologically is

done on guinea pig (Cavia cobaya) since it has high

similarity to human biologically (Legendre, 2016).

2 MATERIAL AND METHOD

This research was approved by the ethics committee

of Biology Department, University of Sumatera

Utara. Thirty guinea pigs weighing 250-400 grams

aged 2-4 month were kept under standard condition 3

days before the research begins and were all in good

condition during the experimental period. Guinea pig

which lost the separator or die before the observation

time is excluded in this research. Guinea pigs were

randomly divided into five groups of 6 animals each:

control group (no treatment), and four experimental

groups (observed on day 7, 14, 21, and 28) and were

then fitted with 0,5 mm Ormco elastomeric separator

in the left maxilla incisors on day 0. The fitted

separator is then removed and replaced with the new

separator in both maxilla incisors on day 7 to produce

continuous force. Six guinea pigs in each group were

euthanized using 75 mg/kg ketamine on days 0, 7, 14,

21, and 28 after the orthodontic force is applied. After

euthanasia, the anterior maxilla with two incisors

were cutted longitudinally and fixed in a 10%

buffered formalin solution for 24 hours. After

washing with water, the specimens were dehydrated

and embedded in paraffin. Slices measuring 4-6 μm

in thickness were obtained and stained with

haematoxylin- eosin. The quantitative analysis of

osteoclast and osteoblast cells is then done by

observing the histologic slide under microscope with

400x magnification. Cells count is done in five field

of view to get the average value in each pressure and

tension side and is done by two observers. Data were

expressed as mean and standard deviation values. The

data’s normality was tested with Saphiro-Wilk and

independent t test was done as an intern rater test. The

data were then analyzed using Repeated

Measurement Test of ANOVA.

Figure 1: Placement of 0,5 mm Ormco elastomeric

separator in the left maxilla incisor on day 0, and the

replacement of new separator in both maxilla incisors on

day 7, 14, and 21 to produce continuous force.

ICOSTEERR 2018 - International Conference of Science, Technology, Engineering, Environmental and Ramiﬁcation Researches

506

Figure 2: Microscopic view of osteoclast and osteoblast cells with 400x magnification.

3 RESULTS

Five guinea pigs were excluded from this study.

Three guinea pigs died and the other two has been lost

for the separator before observation time.

Saphiro Wilk is used as a normality test and

resulting p>0,05 indicates that the data were normally

distributed. Independent t test showed no significant

differences between the observers’ data (p>0,05).

Table 1: Osteoclast Cells Distribution in Five Timesperiod

Observation Time

Mean±SD

Day 0

0,64±0,26

Day 7

1,32±0,77

Day 14

1,20±1,03

Day 21

0,56±0,38

Day 28

0,32±0,36

Table 2: Osteoblast Cells Distribution in Five Timesperiod

Observation Time

Mean±SD

Day 0

7,04±0,36

Day 7

5,80±0,24

Day 14

6,04±0,90

Day 21

6,44±0,74

Day 28

5,04±0,93

Table 3: Osteoclast Quantity Differences of Five

Timesperiod

(Days)

Mean

Differences

Significance

value

0-7

+0,680

1,000

0,061

7-14

-0,120

1,000

14-21

-0,640

1,000

21-28

-0,240

1,000

Table 4: Osteoblast Quantity Differences of Five

Timesperiod

(Days)

Mean

Differences

Significance

value

0-7

-1,240

0,015

0,006

7-14

+0,240

1,000

14-21

+0,400

1,000

21-28

-1,400

0,220

Table 5: Quantity Differences of Osteoclast between

Control and Experimental Groups

Days

Mean

Differences

Significance

0 7

-0,680

1,000

-0,560

1,000

0,080

1,000

0,320

0,028

Table 6: Quantity Differences of Osteoblast between

Control and Experimental Groups

Days

Mean

Differences

Significance

0 7

1,240

0,015

1,000

0,601

0,600

1,000

2,000

0,232

Figure 3: Activities of Osteoclast and Osteoblast

Osteoclast and Osteoblast Quantity Change in Guinea Pig’s Tooth Movement

507

4 DISCUSSION

Figure 3 illustrate the activities of osteoclast and

osteoblast on the pressure and tension side. The

quantity of osteoblast cell found much more than the

osteoclast cells. This result is corresponding with

Sutantyo’s research that state out osteoclast reactivity

is not only based on the quantity, but also the quality

of the cell itself (Patil and Jayade, 2006).

Table 1 and table 2 illustrate the osteoclast and

osteoblast cells distribution in five times period. A

number of osteoclast and osteoblast cell was observed

on day 0, which prove that resorption and deposition

of bone as a remodelling process always happen

although no orthodontic force is given. A research by

Miyoshi et al., state that control samples with no

treatment undergone a physiologic bone remodelling

process showed by the movement of the tooth (Patil

and Jayade, 2006). Table 3 and table 4 illustrate the

osteoclast and osteoblast quantity differences in five

times period. Data analysis shows no significant

differences of osteoclast quantity in five times period

(p=0,061), but there were significant differences of

osteoblast quantity (p=0,006). Table 5 and table 6

illustrate the quantity differences of osteoclast and

osteoblast between control and experimental groups.

Post Hoc test with Bonferroni correction showing

significant differences of osteoclast cell quantity

between control group and the group observed on

days 28 (p=0,028), and significant differences of

osteoblast cell quantity between control group and

group observed on days 7 (p=0,015).

5 CONCLUSIONS

In conclusion, the finding of the present study

show that the activities of osteoclast and osteoblast

cells are not only depends on its quantity, but also its

quality. The present study also concludes that cells’

activities in orthodontic tooth movement reach its

optimum to move the tooth in a cycle of less than 28

days as the osteoclast and osteoblast cells quantity

will drop after days 28.

ACKNOWLEDGEMENTS

We are very extremely privileged this

completion of our projects addressed to our Research

Institution from Universitas Sumatera Utara at the

expense of this study from TALENTA funding in

year 2018 and our very great appreciation for

Hospital of University Sumatera Utara, Animal

House of Biology’s Department and Patology

Anatomy Department from our beloved Universitas

Sumatera Utara.

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