The animals were euthanized by overdose inhalation of 5% isoflurane after 12 days of tooth movement. After dissection of the maxillary region, the distance between the first and second molars was measured. Impressions of the teeth and the maxilla were obtained with the use of individual trays containing hydrophilic vinyl polysiloxane impression material (EXAMIXFINE Injection Type, GC Co., Tokyo, Japan). After an impression was obtained, the samples were fixed in 4% paraformaldehyde. Tooth movement was assessed by measuring the shortest distance (black double arrow) between the distal marginal ridge of the first molar and the mesial marginal ridge of the second molar (dotted line) under a stereoscopic microscope (VH-7000; Keyence, Osaka, Japan) as described previously [28 (link), 40 (link), 41 (link)] (S4 Fig ). The left side of the maxilla was used for evaluation in each of the groups.
Vh 7000
Manufactured by Keyence
Sourced in Japan
The VH-7000 is a high-performance digital microscope designed for laboratory and industrial applications. It features a high-resolution camera and advanced image processing capabilities to capture detailed images and video of small-scale samples.
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Lab products found in correlation
8 protocols using vh 7000
1
Measurement of Tooth Movement in Rodents
2
Analyzing Microscopic Surface Topography
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The contact surface of the tips was observed by optical stereomicroscopy (VH-7000 ® , Keyence, NJ, USA) and scanning electron microscopy (SEM) (S-4500, Hitachi, Tokyo, Japan). Prior to evaluation, the tip surface was rinsed with water spray and dried by an air syringe without any other cleaning or treatment. Then, following sputter coating with osmium plasma at a thickness of 4 nm, the SEM secondary electron image of the tip contact surface was taken at an accelerating voltage of 15 kV.
3
Measuring Tooth Movement in Mice
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The mice were anaesthetised after 12 days of OTM. We measured the space between the first and second molars using a tray containing hydrophilic vinylpolysiloxane (EXAFAST Injection Type, GC Co., Tokyo, Japan) to obtain an impression of the maxillary teeth. We used stereoscopic microscopy (VH-7000; Keyence, Osaka, Japan) to evaluate tooth movement with the closest distance between the distal marginal ridge of the first molar and the mesial marginal ridge of the second molar (Figure 1(b) ).
4
Dental Tissue Analysis Protocol
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On day 14, all experimental animals were sacrificed using intraperitoneal injection of 250 mg/kg pentobarbital sodium. Furthermore, maxilla along with the teeth was resected to measure the central incisors distal movement gap and to prepare the specimens for HSP-70 and MMP-8 expression immunohistochemistry (IHC) examination.
The impression of maxillary central incisors was made to measure the distal movement gap with the use of individual trays containing hydrophilic vinyl polysiloxane impression material (EXAFAST Injection Type, GC Co., Tokyo, Japan). The samples were fixed in 4% paraformaldehyde after the impressions were obtained. The amount of teeth movement was evaluated by measuring the closing distance between the central incisors in the impression under a stereoscopic microscope [VH-7000; Keyence, Osaka, Japan;Figure 2b ]. For each mouse, the measurement was taken four times, and the mean value was used.
For IHC test, collected tissues were blocked with paraffin before being cut and fixed to object glass. The samples were analyzed by IHC staining, using monoclonal-antibody (MoAb) anti-HSP-70 and MoAb anti-MMP-8 (BioRad, Hercules, USA). The results were examined under digital microscope at ×400, equipped with a Nikon microscope OPTIPHOT (Nikon, Tokyo, Japan).
The impression of maxillary central incisors was made to measure the distal movement gap with the use of individual trays containing hydrophilic vinyl polysiloxane impression material (EXAFAST Injection Type, GC Co., Tokyo, Japan). The samples were fixed in 4% paraformaldehyde after the impressions were obtained. The amount of teeth movement was evaluated by measuring the closing distance between the central incisors in the impression under a stereoscopic microscope [VH-7000; Keyence, Osaka, Japan;
For IHC test, collected tissues were blocked with paraffin before being cut and fixed to object glass. The samples were analyzed by IHC staining, using monoclonal-antibody (MoAb) anti-HSP-70 and MoAb anti-MMP-8 (BioRad, Hercules, USA). The results were examined under digital microscope at ×400, equipped with a Nikon microscope OPTIPHOT (Nikon, Tokyo, Japan).
5
Orthodontic Tooth Movement and Space Retention in Mice
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Mice were fit with an orthodontic appliance, as described previously [6 (link), 7 (link)]. Briefly, under anesthesia, a nickel titanium closed coil spring (Tomy; Fukushima, Japan) was fixed between the upper incisors and the upper-left first molar. A 0.1-mm stainless steel wire was then used to move the first molar in a mesial direction. According to the database of the manufacturer, the force level of the appliance after activation is approximately 10 g (S1 Fig ). Orthodontic tooth movement (OTM) was achieved after forced loading for 12 days. The distance between first molar (M1) and second molar (M2) was measured. A tray containing hydrophilic vinylpolysiloxane (EXAFAST Injection Type, GC Co., Tokyo, Japan) was placed onto the maxillary teeth to obtain an impression. The distance between the distal marginal ridge of the first molar and the mesial marginal ridge of the second molar (dotted line) was measured to assess tooth movement (red double arrow) by stereoscopic microscopy (VH-7000; Keyence, Osaka, Japan) (Zaki et al., 2015). Space retention was considered successful when movement was less than 10 μm of the original OTM (S1 Fig ).
6
Tooth Movement Measurement in Rats
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The rats were killed by injecting an overdose of anesthetic (pentobarbital > 25 mg/kg) on the 1st, 3rd, 7th, and 14th days. Three rats were harvested at each point in time. After dissection of the maxillary region, impressions of the teeth and maxillae were obtained with the use of individual trays containing hydrophilic vinyl polysiloxane impression material (EXAFAST Injection Type, GC Co., Tokyo, Japan). The amount of tooth movement was evaluated by measuring the closest distance between the first and the second molars in the impression under a stereoscopic microscope (VH-7000; Keyence, Osaka, Japan).
7
Measuring Orthodontic Tooth Movement
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After 12 days of force loading, the tooth movement space between the first and second molars was determined by taking an impression of the maxillary teeth with hydrophilic vinyl polysiloxane (EXAFAST Injection Type; GC Co., Tokyo, Japan) under deep anesthesia. The distance of tooth movement was measured as the distance between the distal marginal ridge of the left first molar and the mesial marginal ridge of the left second molar as the OTM side, and between the distal marginal ridge of the right first molar and the mesial marginal ridge of the right second molar as control side by stereoscopic micro-scopy (VH-7000; Keyence, Osaka, Japan) as described previously.17 (link)
8
Measuring Tooth Movement in Maxilla
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After tooth movement, tissues from the maxillae were fixed in 4% formaldehyde overnight, and then observed using μ-CT (R_mCT; Rigaku, Tokyo, Japan). The image acquisition conditions were as follows: radiograph source voltage, 90 kV; current, 150 μA; scanning time, 2 min; resolution, 20 μm/pixel. Tooth movement was evaluated by measuring the shortest distance between the first and second molars in the impression under a stereoscopic microscope (VH-7000; Keyence, Osaka, Japan) as in previous reports (n = 7).12 , 13 (link), 14 (link), 15 (link), 16 (link),20 (link),21 (link)
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