Stimulation of bone formation in the expanding inter-premaxillary suture by vitamin E, in rat

Tancan Uysal; Mihri Amasyali; Huseyin Olmez; Yildirim Karslioglu; Omer Gunhan

doi:10.4041/kjod.2009.39.5.337

Journal List > Korean J Orthod > v.39(5) > 1043603

Go to TopGo to Top Go to BottomGo to Bottom

TOOLS

Uysal, Amasyali, Olmez, Karslioglu, and Gunhan: Stimulation of bone formation in the expanding inter-premaxillary suture by vitamin E, in rat

Original Article

Korean J Orthod 2009;39(5):337-347.

Published online: 21 January 2009

DOI: https://doi.org/10.4041/kjod.2009.39.5.337

Stimulation of bone formation in the expanding inter-premaxillary suture by vitamin E, in rat

Tancan Uysal, DDS PhD^a, Mihri Amasyali, DDS MS^b, Huseyin Olmez, DDS PhD^c, Yildirim Karslioglu, MD^d, Omer Gunhan, DDS, PhD^e

^aAssociate Professor and Chair, Department of Orthodontics, Faculty of Dentistry, Erciyes University, Kayseri, Turkey.

^bResearch Assistant, Associate Professor, Department of Orthodontics, Centre of Dental Sciences, Gülhane Military Medical Academy, Ankara, Turkey.

^cAssociate Professor, Department of Orthodontics, Centre of Dental Sciences, Gülhane Military Medical Academy, Ankara, Turkey.

^dAssistant Professor, Professor, Department of Pathology, Faculty of Medicine, Gülhane Military Medical Academy, Ankara, Turkey.

^eProfessor, Department of Pathology, Faculty of Medicine, Gülhane Military Medical Academy, Ankara, Turkey.

Corresponding author: Tancan Uysal. Erciyes Universitesi, Dis Hekimligi Fak. Ortodonti AD, Melikgazi, Kayseri, Turkey 38039. ＋903524380657; e-mail, tancanuysal@yahoo.com.

^* This work was supported by a research grant from Gulhane Military Medical Academy Research Scientific Committee (AR-2009-04).

Received 24 June 200921 September 2009 Accepted 23 September 2009

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Objective

The aim of this study was to evaluate the effects of vitamin E (α-tocopherol) administration on bone formation in response to expansion of the inter-premaxillary suture, in rats, histomorphometrically.

Methods

Thirty 50 - 60 day old Wistar rats were separated into five equal groups (one control and four experimental). All groups were subjected to inter-premaxilla expansion with 50-gram of force. Six control animals received saline solution (Group I) and three experimental groups were treated with a single dose of α-tocopherol injected into the inter-premaxillary suture after one day after appliance placement (Group II: 2 mg/kg; Group III: 10 mg/kg; and Group IV: 50 mg/kg). A further group of six animals received three injections of 10 mg/kg α-tocopherol, one each on days 3, 6, and 9 (Group V). Bone formation in the suture was evaluated by bone histomorphometry. Kruskal-Wallis rank and Mann-Whitney U tests were used for statistical evaluation at p ＜ 0.05 level.

Results

New bone area, bone perimeter, feret’s diameter and newly formed bone measurements were significantly higher in the experimental groups than the control (p ＜ 0.001). Bone architecture in α-tocopherol administrated groups was improved, and bone formation during the expansion period was stimulated significantly, in a dose-dependent manner.

Conclusions

The application of α-tocopherol during the early stages to orthopedically expanded inter-premaxillary suture areas may stimulate bone formation and shorten the retention period, in rats.

Keywords: Vitamin E, α-tocopherol, Expansion, Histomorphometry, Rat

REFERENCES

1.Saito S., Shimizu N. Stimulatory effects of low-power laser irradiation on bone regeneration in midpalatal suture during expansion in the rat. Am J Orthod Dentofacial Orthop. 1997. 111:525–32.

2.Sawada M., Shimizu N. Stimulation of bone formation in the expanding mid-palatal suture by transforming growth factor-β1 in the rat. Eur J Orthod. 1996. 18:169–79.

3.Uysal T., Ustdal A., Sonmez MF., Ozturk F. Stimulation of bone formation by dietary boron in an orthopedically expanded suture in rabbits. Angle Orthod. 2009. 79:984–90.

4.Uysal T., Amasyali M., Enhos S., Sonmez MF., Sagdic D. Effect of ED-71, a new active vitamin D analog, on bone formation in an orthopedically expanded suture in rats. a histomorphometric study. Eur J Dent. 2009. 3:165–72.

5.Ten Cate AR., Freeman E., Dickinson JB. Sutural development: structure and its response to rapid expansion. Am J Orthod. 1977. 71:622–36.

6.Mundy GR. Cytokines and growth factors in the regulation of bone remodeling. J Bone Min Res. 1993. 8:S505–10.

7.Key LL., Ries WL., Taylor RG., Hays BD., Pitzer BL. Oxygen derived free radicals in osteoclasts: the specificity and location of the nitroblue tetrazolium reaction. Bone. 1990. 11:115–9.

8.Göktürk E., Turgut A., Bayçu C., Günal I., Seber S., Gülbas Z. Oxygen-free radicals impair fracture healing in rats. Acta Orthop Scand. 1995. 66:473–5.

9.Turk C., Halici M., Guney A., Akgun H., Sahin V., Muhtaroglu S. Promotion of fracture healing by vitamin E in rats. J Int Med Res. 2004. 32:507–12.

10.Durak K., Sonmez G., Sarisozen B., Ozkan S., Kaya M., Ozturk C. Histological assessment of the effect of alpha-tocopherol on fracture healing in rabbits. J Int Med Res. 2003. 31:26–30.

11.Durak K., Bilgen OF., Kaleli T., Tuncel P., Ozbek R., Turan K. Antioxidant effect of alpha-tocopherol on fracture haematoma in rabbits. J Int Med Res. 1996. 24:419–24.

12.Yilmaz C., Erdemli E., Selek H., Kinik H., Arikan M., Erdemli B. The contribution of vitamin C to healing of experimental fractures. Arch Orthop Trauma Surg. 2001. 121:426–8.

13.Burton GW., Ingold KU. Vitamin E as an in vitro and in vivo antioxidant. Ann N Y Acad Sci. 1989. 570:7–22.

14.Burton GW., Traber MG. Vitamin E: antioxidant activity, biokinetics, and bioavailability. Annu Rev Nutr. 1990. 10:357–82.

15.van Tits LJ., Demacker PN., de Graaf J., Hak-Lemmers HL., Stalenhoef AF. Alpha-tocopherol supplementation decreases production of superoxide and cytokines by leukocytes ex vivo in both normolipidemic and hypertriglyceridemic individuals. Am J Clin Nutr. 2000. 71:458–64.

16.Manolagas SC., Jilka RL. Bone marrow, cytokines, and bone remodeling. Emerging insights into the pathophysiology of osteoporosis. N Engl J Med. 1995. 332:305–11.

17.Turan B., Can B., Delilbasi E. Selenium combined with vitamin E and vitamin C restores structural alterations of bones in heparin-induced osteoporosis. Clin Rheumatol. 2003. 22:432–6.

18.Turan B., Balcik C., Akkas N. Effect of dietary selenium and vitamin E on the biomechanical properties of rabbit bones. Clin Rheumatol. 1997. 16:441–9.

19.Rasband WS. Image-J. U.S. National Institutes of Health, Bethesda, Maryland, USA. http://rsb.info.nih.gov/ij/,1997-2008.

20.Graber TM., Vanarsdall RL., Vig KWL. Orthodontics: current principles and techniques. 4th ed.St. Louis: Elsevier Mosby;2005. p. 257–62.

21.Haas AJ. The treatment of maxillary deficiency by opening the midpalatal suture. Angle Orthod. 1965. 35:200–17.

22.Arjmandi BH., Juma S., Beharka A., Bapna MS., Akhter M., Meydani SN. Vitamin E improves bone quality in the aged but not in young adult male mice. J Nutr Biochem. 2002. 13:543–9.

23.Arjmandi BH., Akhter MP., Chakkalakal D., Khalil DA., Lucas EA., Juma S, et al. Effects of isoflavones, vitamin E, and their combination on bone in an aged rat model of osteopenia. J Bone Min Res. 2001. 16:S533.

24.Swennen G., Dempf R., Schliephake H. Cranio-facial distraction osteogenesis: a review of the literature. Part II: experimental studies. Int J Oral Maxillofac Surg. 2002. 31:123–35.

25.Storey E. Tissue response to the movement of bones. Am J Orthod. 1973. 64:229–47.

26.Hagiwara T., Bell WH. Effect of electrical stimulation on mandibular distraction osteogenesis. J Craniomaxillofac Surg. 2000. 28:12–9.

27.Eriksen EF., Axelrod DW., Melson F. Bone histology and histomorphometry. In: Bone histomorphometry. New York: Raven Press;1994. p. 33–8.

28.Burstone CJ., Shafer WG. Sutural expansion by controlled mechanical stress in the rat. J Dent Res. 1959. 38:534–40.

29.Cope JB., Samchukov ML. Mineralization dynamics of regenerate bone during mandibular osteodistraction. Int J Oral Maxillofac Surg. 2001. 30:234–42.

30.Sencimen M., Aydintug YS., Ortakoglu K., Karslioglu Y., Gunhan O., Gunaydin Y. Histomorphometrical analysis of new bone obtained by distraction osteogenesis and osteogenesis by periosteal distraction in rabbits. Int J Oral Maxillofac Surg. 2007. 36:235–42.

31.Garrett IR., Boyce BF., Oreffo RO., Bonewald L., Poser J., Mundy GR. Oxygen-derived free radicals stimulate osteoclastic bone resorption in rodent bone in vitro and in vivo. J Clin Invest. 1990. 85:632–9.

Fig 1.

Appliance in situ.

Fig 2.

Occlusal radiographs. A, Baseline; B, end of expansion; C, end of retention.

Fig 3.

Haematoxylin-eosin (HE) staining of the histological sections prior to optical-microscope examination (× 40 magnification).

Fig 4.

Histomorphometric measurements of newly formed bone area (μm²). Left, Image enhancement and outlining of the area of interest, which is newly formed bone. Histomorphometric measurements of newly formed bone (percentage): Right, marking the intersections of the grid just over the newly formed bone and non-osseous connective tissue by mouse-clicking. At the end of this macro, the program gives a percent indicating the ratio of bone to the connective tissue.

Fig 5.

Photomicrograph of a section in the expansion area of the control group showing that it appears to be occupied by fibrous tissue, but not the trabecular bone. Large connective tissues indicates beginning stages of bone formation (HE 200 × magnification) (bar = 30μm).

Fig 6.

Photomicrograph of a section in the expansion area of the experimental group (Group IV) showing larger masses of new bone trabeculae. New bone became attached to old bone at the site of expansion. Large amounts of new bone forming area indicates later stages of bone formation (HE 200 × magnification) (bar = 30μm).

Table 1.

Body weight changes (kg) between groups during expansion and retention periods

Groups	N	T1-T0		T2-T0		Significance
Groups	N	Mean S	Standard deviation	Mean Staandard	deviation	T1-T0	T2-T0
Group I	6	0.319	0.195	0.337	0.122	NS	NS
Group II	6	0.303	0.148	0.322	0.179	NS	NS
Group III	6	0.289	0.137	0.325	0.146	NS	NS
Group IV	6	0.291	0.127	0.312	0.139	NS	NS
Group V	6	0.275	0.156	0.305	0.123	NS	NS

T0, baseline weight; T1, end of expansion (5-day) weight; T2, end of retention (10-day) weight; NS, not significant.

Table 2.

The amount of expansion of each group and biometric analysis (unit: μm)

Groups	N	Mean	Std. deviation	Minimum	Maximum	Significance
Group I	6	338.23	19.79	308.87	365.63
Group II	6	330.59	22.73	314.71	360.95
Group III	6	317.35	24.79	297.17	355.38	NS
Group IV	6	312.78	22.27	287.35	346.98
Group V	6	318.11	21.14	300.72	342.65

Group I, Control; Group II, 2 mg/kg α - tocopherol; Group III,10 mg/kg α - tocopherol; Group IV, 50 mg/kg α - tocopherol; Group V, 10 mg/kg α - tocopherol at days of 3, 6, and 9.

Table 3.

Descriptive values, Kruskal Wallis and Mann-Whitney U results of histomorphometric measurements (unit: μm)

Parameters	Groups	N	Mean	SD	Min	Max	F value	Significaance	Multiple comparisons (Mann-Whitney U test)
Parameters	Groups	N	Mean	SD	Min	Max	F value	Significaance	Group II	Group III	Group IV	Group V
Area (μm²)	Group I	6	66,755	13,449	48,719	89,570			†	‡	‡	‡
	Group II	6	84,725	9,148	69,876	95,674				‡	‡	‡
	Group III	6	125,687	12,009	110,545	140,400	136,327	‡			‡	NS
	Group IV	6	139,117	10,322	123,435	154,173						*
	Group V	6	127,324	10,620 1	112,545	144,005
Perimeter (μm)	Group I	6	134,649	13,362	115,723	154,773			†	‡	‡	‡
	Group II	6	154,653	9,120	139,841	165,545				‡	‡	‡
	Group III	6	187,090	8,116	175,178	195,907	126,239	‡			‡	NS
	Group IV	6	254,474	7,522	235,478	252,796						‡
	Group V	6	185,424	5,911	175,178	191,866
Feret's diameter (μm)	Group I	6	14,933	8,128	12,083	17,148			‡	‡	‡	‡
	Group II	6	57,212	14,332	42,865	76,111				NS	‡	NS
	Group III	6	74,840	11,423	59,690	88,988	29,612	‡			NS	NS
	Group IV	6	94,840	14,283	88,988	99,690						NS
	Group V	6	73,173	9,608	59,690	97,680
Newly formed bone(%)	Group I	6	35,365	12,947	20,000	55,550			‡	‡	‡	‡
	Group II	6	62,165	8,439	53.33	76,340				NS	*	NS
	Group III	6	72,055	7,086	63,740	80,840	22,256	‡			NS	NS
	Group IV	6	77,721	4,923	73.74	86,660						NS
	Group V	6	72,721	8,664	63,740	84,840

SD, standard deviation; Min, minimum; Max, maximum; NS, not significant; * p＜ 0.05, † p ＜ 0.01, ‡ p＜ 0.001; Group I, control; Group II, 2mg/kg α-tocopherol; Group III, 10mg/kg α-tocopherol; Group IV, 50mg/kg α-tocopherol; Group V, 10mg/kg α-tocopherol at days of 3, 6, and 9.

TOOLS

Similar articles