Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
51
EFFECT OF LOW LEVEL LEASER THERAPY ON THE
CHRONIC DEFECT OF TIBIAL BONES IN RABBITS
Humam H.Nazht ,Sinan A.N. Al-khazrajii, Raffal A.Omar
Department of Surgery and obstetric ,College of veterinary medicine, University of
Baghdad,Baghdad ,Iraq.
Keys words: LLLT , Chronic bone defect,Tibial bone..
Corresponding Author;[email protected]
ABSTRACT
This project was designed to evaluate the effect of low level leaser therapy
(LLLT) on the induced chronic defect in the tibial bone of rabbits radiographically ,
and physically,eight adults local breed rabbits of both sexes were employed to
induced 2.5 mm in diameter circular defect in the tibial bone surgically under general
anesthesia and highly aseptic technique ,the induced hole was implanted with 2.5 mm
stainless steel screw ,that removed from the bone after one and half month post
operation , the experimental animals randomly divided into two equal groups four
animas of each, the control group was lifted for normal healing process of the bone
defect without any treatment ,while the treatment group was exposed daily with single
dosage of continuous diode laser for five minutes of 850 nm and 148.4 j/cm2 at 72
hours interval for 14 days post operation (p.o.) ,the radiographic results in the
treatment group showed increase in the bone density around the induced defect with
new bone formation fill the cavity at the end of 2nd week p.o. then increase in the
bone density and volume at the end of 4th week p.o. compare with the control group
that the density and the new bone formation less than treatment group ,the physical
analysis revealed increase in the bone density and hardiness with significant increase
in fractures tolerance in the treatment group compare with the control group .In
conclusion that the LLLT successfully used for repairing the critical ( chronic) defect
in tibial bones in rabbits .
INTRODUCTION
Chronic bone defect can be induced by removing the stainless steel screw
implantation one month or more p.o. especially in the dentist researches, or after
removing the internal and external fixation methods of bones fractures, these circular
bone defect may be more or less than 2.5 mm ( authors opinion), one of the best
treatment methods of these defect are by bone grafts/implantation (1- 4) ,or even by
laser therapy (5). laser can be used successfully for the repair of bone defect (6 the
author himself ) in which I refer in my work that laser therapy has appositive effect on
the treatment of the bone defect of the lower mandible of rabbits ,that laser stimulated
osteoblast cells. laser can be used strongly and successfully to promote fracture
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
52
healing (7-9),in which the mechanisms of laser can be done by increase bone
metabolism ,stimulation of collagen synthesis , angiogenesis ,and it increase cells
proliferation and differentiation of the mesynchamel cells , the increase osteoblast
stimulation leads to osteon production and deposition with increase calcium and
minerals deposition (10 and 11).
Low-power lasers therapy which (then called laser biostimulation) appeared
before 1970s when Ender Mester trying to study the carcinogenic effect of laser on
mice (12 and 13) . Several in vitro and in vivo studies was done to evaluate the effect
of laser on different cells (13).The aims of Laser therapy were to restore the normal
biological function of injured or stressed cells (14). Many workers refer that laser can
be used in acceleration of wound healing, acceleration remodeling and bone repair,
restoration of normal neural function following injury, normalization of abnormal
hormonal function, pain attenuation, stimulation of endorphin release, and modulation
of the immune system (15) ,The indications of laser therapy are presented on both soft
and hard tissues (16).Laser can be used for medical and Non- medical purposes, the
most important applications are in the areas of major medical surgery, cosmetic and
plastic surgery, dentistry and ophthalmology (17 ).
The mechanism of mode of action of laser as reported by (14) the laser photonic
energy is absorbed by the mitochondria in the cell in which is converted to chemical
kinetic energy and finally leads to more production of ATP. ATP is the source of
energy in the cell which is necessary for cell activities such as synthesis of DNA,
RNA and proteins that are important in cellular proliferation.
MATERIALS AND METHODS
Eight adults' local breed rabbits of both sexes were employed for induced hole
of 2.5 mm diameter in the medial aspect of tibial bone ,using the electrical drill under
general anesthesia by intramuscular injection 17 mg /Kg .B.W. 2% xylaxine
hydrochloride after 10 minutes 37mg /Kg .B.W. 10% ketamine hydrochloride
respectively, and to avoid thermal reactions by cooling the bone with dripping locally
sterile cold normal saline during the operation ,the induced pore were implanted with
suitable size of stainless steel screw for 1 month and half p.o.( Fig.1 A) , then the
screw removed surgically and the animals divided to two equal group each has 4
rabbits ( Fig.1 B) ,the control group lifted without any treatment ,while the treatment
group exposed to single daily dose of continuous diode laser at the medial side with a
dose of (850 nm /148.4 J/cm for 5 minutes at 72 hours interval for 14 days).Daily
observation for both group for 1 month after removing the stainless devices (two
months and half p.o.). All the animals were radiographically examined at 15th and 30th
days after removing the stainless steel screw, they scarified by high dose of
anesthesia, the bone specimen sent for the physical lab analysis to estimate the bone
density, bone hardiness, and fractures tolerance (physical laboratory of the al-nahreen
university).
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
53
RESULTS AND DISCUSSION
The radiographic finding of the treatment group 15th days after laser therapy
revealed increase in bone density around the induced chronic defect (Fig.2 A)
,compare with the control group which lifted without laser irradiation ,the area around
the induced defect shown less density ( Fig.2 B),and at the end of the one month's
post irradiation the treatment group shown that the cavity filled with the new bone
formation with the increase of the bone density around the induced hole( Fig.3 A),
while in the control group the new bone formation and the bony density less than
treatment group ( Fig.3 B),and this is due to osteoblast stimulation by laser therapy
which lead to osteon production and deposition ,as well as increase mineral and
calcium deposition (6,10,and 11).and this knots is consider with (18),that the a
sclerotic area which appears high radiographic density is due to osteoblast stimulation
and osteod formation and increase mineralization.
the high radiographic density around the induced critical defect with the new bone
formation in the treatment group which appears more clear than the control group is
due to effect of Laser therapy, which can used in accelerate fracture healing by
increase in callus volume and mineralization due to osteoblast activity especially in
early stages of healing process ( 10,19, and 20).
All the physical analysis shows increase in the bone density ( Fig. 4) and
hardiness ( Fig. 5)of the exposed group with LLLT ,compare to the control group and
this due to positive effect of laser therapy on osteoblast cells stimulation with osteon
production and deposition and all these mentioned by (6,10, and 11). The increase
resistance of bone fracture, in the treatment group compare with the control group
which lifted without irradiation (Fig.6). Was because of the laser therapy ,which result
in increase hardiness due to calcium and minerals deposition (10 and 11),and these
statement agree with others authors which they mentioned that exposed to laser
therapy can increase the amount of new bone formation and increase mineralized at
the new bone tissue because of the laser stimulation on the osteblast cells (20) ,and
these finding also agree with (18),that the hardness of the bone is due to the osteoblast
stimulation and calcium deposition .
Laser accelerate, bone fracture healing by accelerating callus tissues formation and
bone mineralize, due to osteoblastic activity, it enhance callus development in the
early stage of the healing process, with improvement in biomechanical properties of
the healing bone (10, 19, and 20).Many researchers declare that low intensity Laser
irradiation in the early stage stimulating mineralization of new trabecular bone
formation in surgically created bony defects (21 and 22).
CONCLUSION
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
54
LLLT can be used for the treatment of tibial critical (chronic) defect or cavities.
Fig. 1 A. implantation of the stainless steel screw remain for one and half month p.o.
B. Remove the stainless steel screw after one and half month's p.o.
Fig. 2 after 15th days post laser irradiation. Shown increase of bone density with new
bone formation inside the bone defect more obvious in the treatment group A.
than the control group B.
A B
A B
A B
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
55
Fig.3 after one month after irradiation. A sclerotic area around the induced hole
showed high radiographic density and fill the empty defect with new bone
formation in the treatment group A. compare with the control group B.
Fig. 4 increase bone density of the treatment group compare with the control group
less and unstable.
Fig.5 increase of bone density of the treatment group compare with the control group
which show less and unstable
Fig .6 fracture tolerance in the treatment group more the control group
2.3
2.5
2.7
2.9
3.1
3.3
3.5
3.7
1000 1050 1100 1150 1200 1250 1300
Bulk Density
Temperature (oC)
before
after
0
2
4
6
8
1000 1050 1100 1150 1200 1250 1300
Vickers Hardness
Temperature (oC)
before (Vickers Hardness)
0
0.2
0.4
0.6
0.8
1
1.2
1000 1050 1100 1150 1200 1250 1300
Fracture Toughness Temperature (
oC)
before (Fracture Toughness)
after
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
56
تاثیر اللیزر الواطئ الطاقة على الاصابة المزمنة فی عظام القصبة فی الارانب
ھمام حسام الدین محمد نزھت، سنان عدنان محمد نوری الخزرجی،رفل عبد الدایم عمر
فرع الجراحة والتولید البیطری, کلیة الطب البیطری ,جامعة بغداد,العراق
الخلاصة
صمم ھذا البحث لدراسة تاثیر استخدام اللیزر واطئ الطاقة على الاذى المحدث فی عظم القصبة فی
الارانب.استخدمت ثمانیة ارانب ناضجة محلیة من کلا الجنسین لاحداث ثقب دائری بقطر ٢.٥ ملم فی عظم
القصبة جراحیا تحت التخدیر العام وباستخدام شروط التطھیر والتعقیم المتعارف علیھا ،وذلک بزرع قطعة من (
بقطر ٢.٥ ملم ثم تم رفعھا بعد شھر ونصف من العملیة .تم تقسیم حیوانات التجربة الى (stainless steel
مجموعتین متساویة کل مجموعة تحوی اربع ارانب . مجموعة السیطرة ترکت للمراقبة بدون ایة تعریض ،اما
مجموعة العلاج فقد تم تعریض منطقة العملیة الى جرعة واحدة یومیا من اللیزر واطئ الطاقة نوع اللیزر
المستمر لمدة خمسة دقائق بطاقة ١٤٨.٤ جول /سم ٢ بطول موجی ٨٥٠ نانو میتر کل ٧٢ ساعة لمدة ١٤ یوم من
التعریض.واستمرت المتابعة لمدة شھرین ونصف من العملیة واستخدمت الفحوصات التالیة .الفحوصات
الاشعاعیة للفترات اسبوعین واربع اسابیع بعد التعریض.والفحوصات الفیزیائیة لقیاس مستوى الکثافة والصلادة
وقابلیة تحمل الکسر لمنطقة التعریض باللیزر .واظھرت نتائج الفحوصات الاشعاعیة بظھور کثافة عظمیة فی
منطقة التعریض حول الثقب للفترة اسبوعین بعد التعریض واستمرت کثافة العظم المتکون حول وداخل الثقب
مقارنة مع مجموعة السیطرة الذی لم یکن بالکثافة والوضوح فی مجموعة العلاج .
اما نتائج الفحوصات الفیزیائیة فقد بینت وجود زیادة فی کثافة العظم والصلادة مع زیادة واضحة فی قابلیة
التحمل للکسر فی منطقة التعریض فی مجموعة العلاج مقارنة مع مجموعة السیطرة .
نستنتج من ھذة الدراسة ھو ان اللیزر واطئ الطاقة ممکن استخدامة فی علاج الاصابات المزمنة المستحدثة فی
عظام القصبة فی الارانب
REFERENCES
1. Matsumine, A.; Myoui, A.; Kusuzaki, K.; Araki, N.; Seto,M. and Yoshikawa,H.et
al., (2004).Calcium hydroxyapatite ceramic implants in 5.bone tumour surgery.A
long-term follow-up study. J Bone Joint Surg Br; 86:719–25.
2. Ogose, A.; Hotta, T.; Kawashima, H.; Kondo, N.and Gu W, Kamura T, et al.
(2005) .Comparison of hydroxyapatite and beta tricalcium phosphate as bone
substitutes after excision of bone tumors .J Biomed Mater Res B Appl
Biomater;72:94–101.
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
57
3. Walsh, WR.; Vizesi, F.; Michael, D.; Auld, J.; Langdown, A. and Oliver, R. et al.
(2008) .Beta-TCP bone graft substitutes in a bilateral rabbit tibial defect model.
Biomaterials; 29:266–71.
4. Nazht, H.H.; Omar, R.A. and Ahmed, H.K.( 2018a): Sheep rib xenograft femoral
bone implant in rabbits OJVR .Volum22 (4) : 290,299.
5. Hallman, M. and Thor, A. (2008). “Bone substitutes and growth factors as an
alternative/complement to autogenously bone for grafting in implant dentistry,”
Periodontology. 47, (1): 172–192.
6. Nazht, H.H.(2013).Histopathological study of the effect of the laser on the
osteoblast cells during mandible defect in rabbits, AL-Anbar J.Vet. Sci.,Vol.6
No. (2).
7. Nazht, H. H. ; Faleh I. B.and Hamed N.A. (2016). Histopathological Study of the
Distal Third Fractures of Radius in Dogs, Treated with LLLL,
Bas.J.Vet.Res.Vol.15, No.1.
8. Nazht ,H. H. and Hamed,N. A. (2017) .Radiological Effect of the Low Level
Laser Therapy on Fracture Healing in the Distal Third of Dogs. Elixir Hor. & Sig.
107 42209-42211
9. Nazht, H.H.; Omar, R.A.; AlDahan, R.A;and Ahmed H.K.( 2018b). Effect of
Low Level Laser Therapy on the Sheep Ribs Xeno Graft in the Treatment of
Rabbits Long Bone Fractures. Global Proceedings Repository, American
Research Foundation. Available online at http://proceedings.sriweb.org . The
Ninth International Scientific Academic Conference. Under the Title
“Contemporary trends in social, human, and natural sciences”
10. Shakouri, S K.;Soleimanpour, J.;Salekzamani, Y.and Oskuie,M. R.(2010). Effect
of low-level laser therapy on the fracture healing process. Lasers, Volume
25, Issue 1, pp 73-77.
11. Merli, L.A.; V. P. de Medeiros, V.P. and Toma, L. (2012). “The low level laser
therapy effect on the remodeling of bone extracellular matrix,”Photochemistry
and Photobiology.88, (5):1293–1301.
12. Michael, R.; Hamblin, Tatiana N.;and Demidova. (2006) Mechanisms of low
level light therapy. proc. Of SPIE; 6140,614001.
Basrah Journal of Veterinary Research,Vol.17, No.3,2018
Proceeding of 6th International Scientific Conference,College of Veterinary Medicine
University of Basrah,Iraq
58
13. Blaya, D. S.;Pozza,D.H.;João Batista Blessmann Weber,J.B.B.; de Oliveria,M.G.
(2008) Histological study of the effect of laser therapy on bone repair. The
journal of contemporary dental practice, 9 (6)
14. Hawkins. and Abrahamse.(2007).Phototherapy-a treatment modality for wound
healing and pain relief. African Journal of Biomedical Research. 10:99-109.
15. Walsh L.J. (1997) A current status of low level laser therapy in dentistry. Part I
soft tissue applications. Australian Dental Journal; 42:4
16. Carmen D. M. Todea. (2004).Laser application in conservative dentistry. TMJ;
54:(4)
17. Jan Bone. (2008).Opportunities in laser technology careers, ch.3 pp. 34-46 Mc
Graw Hill.
18. Marsell, R.and Einhorn, TA. (2011).The biology of fracture healing, Injury;
42:551–555.
19. Prado-Filho, J.C. and Sterman, F.A. (2004). “Evaluation of mineral bone
density in thoroughbred horses under an initial trainningprogramme,” Brazilian
Journal of Veterinary Research Animal Science, 41(6): 384-388.
20. Lirani-Galvão, AP.; Jorgetti, V.' da Silva, OL. (2006).Comparative study of how
low-level laser therapy and low-intensity pulsed ultrasound affect bone repair in
rats. Photomed Laser Surg.;24(6):735-40.
21. Dortbudak, O.; Haas, R.; and Mallath-Pokorny G.(2000).Biostimulation of bone
marrow cells with a diode soft laser. Clin Oral Implants Res.Dec;11(6):540-545.
22. . Cerqueira, A. ; Silveira, RL. ; Oliveira, MG. de. ;and
Sant’anaFilho M and Heitz C: (2007). Bone tissue microscopic findings related to
the use of diode laser (830 nm) in ovine mandible submitted to distraction
osteogenesis. Acta Cir Bras 22: 92-97..
