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Low Level Laser Therapy for Healing of Aural Haematoma in Dogs.

Abstract

The study was conducted to evaluate the efficacy of low level laser therapy on healing of aural haematoma in twelve (12) dogs. These dogs were subjected to various clinical examinations viz. degree of inflammation, exudation, embedding of sutures and gross appearance of suture line and recorded on 1st, 3rd, 5th, 7th and 9th day post-operatively using visual score card method. Haemato-biochemical estimations were performed prior to surgery (day 0) and subsequently on 7th day post-operatively. Laser treated dogs revealed less inflammation, exudation, minimum embedding of sutures along with faster healing. The haematological and biochemical parameters remained within normal range by laser treatment, suggestive of no adverse effect on vital body functions, making it a safer therapeutic modality free from any side effects.

Keywords: Aural haematoma; dog; low level laser therapy

Introduction

Aural haematoma occurs most frequently in dogs having pendulous ears. Haematoma is generally recorded on concave surface of ear, however occasionally it can be observed on convex surface or on both sides (Kumar, 2008). Primary healing of sutured wounds takes place in 8-12 days, however complete wound healing takes place in 3 weeks.To reduce this period of healing of surgical wounds and to avoid post-operative complications like excessive exudation, inflammation, oedema, embedding of suturesoccasional scarring and pinnal distortion (Joyce, 2000), low level laser therapy was evaluated post-operatively in the present study.

Materials and Methods

The present study was undertaken on 12 dogs brought for treatment of aural haematoma. The dogs were randomly divided into three groups, each group consisting of four dogs each. Before starting surgical procedure, complete physical examination of all dogs was carried out to ensure their physical status and complete history of each patient was recorded including age, sex, breed, involvement of area of haematoma, duration of haematoma and probable etiological factors.

Routine operation of haematoma was carried out in each dog under premedication with Atropine sulphate @ 0.04 mg/kg and Xylazine hydrochloride @1.5 mg/kg b.wt. I/M followed 10 min later by Ketamine hydrochloride @ 6 mg/kg b.wt. I/M as general anesthetic. Surgery was performed by giving longitudinal incision on inner aspect of ear pinna in centre of swelling, depending on site of haematoma. After proper curetting of both the edges of ear pinna, series of interrupted mattress sutures were applied using mersilk as suture material with knots on outer aspect of ear. Daily post-operative dressing and laser therapy was done as described for different group till healing of the wound.

The group I was treated as control group and antiseptic dressing of surgical wound was carried out with Povidone iodine and local application of Polymixin B sulphate, Neomycin sulphate and Zinc Bacitracin ointment, Inj. Cefotaxim @ 25 mg/kg b.wt. I/M bid for 7 days and Inj. Meloxicam @ 0.3mg/kg b.wt. I/M od was administered for first 3 days.

In group II, dressing of wound + antibiotic and analgesic were as in group I, followed by treatment with low level laser therapy (LLLT) for 7 days continuously for 4 min on convex surface of ear pinna with frequency of 30 Hz and 2.4J of energy.

In group III, dressing of wound + antibiotic and analgesic were as in group I and low level laser therapy was carried out for 7 days continuously for 2 min each on convex and concave surfaces of ear pinna with frequency of 30 Hz and energy of 2.4J. After surgical procedure, these dogs were subjected for various clinical examinations viz. degree of inflammation, exudation, embedding of sutures and gross appearance of suture line, which were recorded on 1st, 3rd, 5th, 7th and 9th day postoperatively using visual score card method in control and laser treated groups.

For estimation of haemato-biochemical parameters, approximately 5 ml of blood was collected aseptically from saphenous or cephalic vein from animals of all three groups. Out of 5 ml, 3ml was collected in EDTA coated vaccutainer vials and 2ml was collected without anticoagulant prior to the surgery (day 0) and subsequently on 7th day post-operatively. Various haemato-biochemical attributes namely total erythrocyte count (million/[micro]l), total leukocyte count (thousand/[micro]l), differential leukocyte count (%), haemoglobin (g/dl), packed cell volume (%), total serum protein (g/dl), serum albumin (g/dl), serum globulin (g/dl), plasma fibrinogen (g/dl), blood glucose (mg/dl) and serum creatinine kinase (U/L) were recorded prior to surgery (day 0) and subsequently on 7th day post-operatively. The haematological attributes were analyzed using haematological analyzer, while serum was used for estimation of biochemical attributes using biochemical analyzer. The data was analyzed by analysis of variance using hierarchical design.

Results and Discussion

Higher incidence was observed in German shepherd breed followed by non descript breeds. The higher incidence of aural haematoma in German shepherds may be because of more number of German shepherds recorded and their more population in and around Jabalpur city. Larsen (1968) also reported higher incidence of aural haematoma in German shepherd and Poodle breed of dogs.

In the present study, higher incidence was observed in dogs between 4-6 years of age followed by 7-10 years. This may be because of large population of this age group of dogs as compared to other age groups. Joyce (2000) also reported higher incidence of aural haematoma in dogs having age between 6-8 years.

In the present study, higher incidence was observed in male as compared to female dogs, which may also be due to more number of male as compared to female and also because of their more activeness. Larsen (1968), also reported higher incidence in male than females.

Incidence of aural haematoma depending upon etiological factor showed ectoparasitic infestation as a main cause followed by otitis externa. In the present study, the higher incidence of aural haematoma because of ectoparasitic infestation may be because of excessive pawing and rubbing of head against the ground due to irritation caused by presence of ectoparasites on body, thereby causing injury to blood vessels of ear resulting into development of haematoma.

In most of cases of aural haematoma, involvement of full ear was observed followed by 3/4th portion of ear. Once the injury take place to blood vessels of ear, collection of fluid starts between cartilage and skin resulting into gradual separation between cartilage and skin and development of haematoma. This further causes discomfort to animal, due to which shaking and pawing of ear increases, resulting in further injury and accumulation of fluid involvement full ear.

In dogs of all three groups, moderate to marked inflammation was observed on 1st day after operation. On 3rd day, mild to moderate inflammation was observed in group I dogs (Fig.1) and in group II and group III, in some dogs it was mild, while in others no inflammation was observed. From 5th day onwards, no inflammation was observed in any of the dog of group II and group III (Fig. 2 and 3), whereas in group I inflammation subsided from 7th day onwards. Less inflammation was observed in laser irradiated group II and III as compared to group I dogs (Table 1), this may be due to anti-inflammatory effect of laser therapy, as also reported by many workers and decrease in plasma level of pro-inflammatory cytokines. The decrease in inflammation by use of laser therapy was also reported by Singh et al. (2011) in dogs, Jann et al. (2012) in equines, Bhowmick et al., (2013) in calves and Hodjati et al. (2014) in rabbits.

The dogs of laser treated group II on 3rd day showed mild to moderate exudation while only mild exudation was observed in group III whereas moderate to marked exudation was recorded in dogs of group I. No exudation was observed in group II and III dogs from 5th day onwards, whereas it was mild till 7th day in group I. More exudation was observed in group I dogs followed by group II and III (Table 1). The decrease in exudation in dogs of groups II and III in ourstudy may be attributed to increased phagocytosis and neo-vascularization at sites of wound because of laser treatment. Decrease in exudation in wounds by use of laser was also reported by Calin et al. (2010) in rabbits, Jovanovi et al. (2010) in equines, Singh et al. (2011) in dogs and Bhowmick et al. (2013) in calves.

In group I dogs on 5th and 7th days, mild to moderate embedding was observed which increased to moderate to marked on 9th day, whereas in laser irradiated II and III groups, no embedding was observed up to 5th day, while on 7th and 9th day, only mild embedding was observed (Fig. 2 and 3). More embedding of sutures was observed in group I dogs as compared to group II and III (Table 1), which may be because of more inflammation for longer period resulting into cutting of skin edges and embedding of sutures (Fig. 1). In the laser treated group II and III, less inflammation was observed at different time intervals, resulting into less cutting or no cutting of skin edges due to which only mild or no embedding was observed.

Overall gross observation revealed that in laser treated group II and III dogs along with less inflammation, exudation and embedding of sutures, showed faster healing, less distortion of ear along with less pendulous nature of operated ear. These findings in our study may be because of less formation of fibrous tissue in group II and III dogs treated with laser therapy, resulting into less distortion of ear. Due to less formation of fibrous tissue, erection of some portion of ear was observed in some dogs of group II and III. Similarly, in laser treated group II and III, no scar formation was observed, while it was present in group I dogs. Faster healing with no scar formation was observed in group II and III dogs, may also be due to influence of laser on healing of wounds. The similar finding with no formation of scar in healing of laser treated wounds was also reported by Souil et al. (2001), Singh et al. (2011) and Bhowmick et al. (2013) in rats, dogs and calves, respectively.

In our study, comparatively high values of total erythrocyte count was recorded on 7th post-operative day, in all three groups (Table-2), which may be due to higher demand of oxygen during tissue regeneration process, as also reported by Calin et al. (2010) in rabbits. The increase in total erythrocyte count also be due to generalized activation of sympatho adernal mechanism, resulting into release of catecholamines which causes spleenic contraction, thereby causing outflux of RBC's in blood circulation, which may be responsible for causing increase in number of total erythrocyte count. Rocha et al. (2009) in rats, Singh et al., (2011) in dogs and Bhowmick et al., (2013) in calves also reported increase in total erythrocyte count in laser treated groups.

The values of total leukocyte count were within normal range at different time intervals in all three groups of dogs (Table-2), as well as between groups, suggestive of no influence of laser therapy on total leukocyte count. Similar findings were also observed by Pallotta et al. (2012) in rats and Bhowmick et al. (2013) in calves.

Haemoglobin concentration showed non-significant increase from 0-7th days interval, however values fluctuated within normal range (Table-2). The higher values of haemoglobin in laser treated group II and group III dogs may be due to higher demand of oxygen during tissue healing process, as faster healing was also observed during repair of haematoma in dogs, irradiated with low level laser therapy. Similar findings were reported by Calin et al. (2010), Singh et al. (2011) and Bhowmick et al. (2013) in rabbits, dogs and calves respectively.

The values of packed cell volume increased non-significantly from 0-7th days interval in all three groups of dogs, but values were within normal range (Table-2), which may be due to no influence of laser therapy on packed cell volume. Similar findings were also reported by Singh et al. (2011) in dogs and Bhowmick et al. (2013) in calves.

Non-significant increase was observed in neutrophil count in group I dogs on 7th day interval, whereas non-significant decrease was observed in group II animals. In group III, dogs significant decrease was observed on 7th day in comparison to 0 day value (Table-3), this may be because of anti-inflammatory property of LLLT, which may be responsible for causing reduction in neutrophil infiltrate. Anvari et al. (2011) in mice also reported significant decrease in value of neutrophil in laser treated wounds as compared to control.

In group I dogs, no change in values of lymphocyte count was observed, while in dogs of group II and group III treated with laser, non-significant increase from 0-7th days was observed (Table-3). In laser treated groups of dogs, this may be due to stimulation of immune system and enhancement of regenerative process by application of laser. Similar findings were also reported by Calin et al. in rabbits (2010) and Singh et al. (2011) in dogs.

Monocyte and eosinophil percent showed nonsignificant change within intervals and between three groups of dogs and values fluctuated within normal range (Table-3). Similar findings were also reported by Singh et al. (2011) in dogs and Bhowmick et al. (2013) in calves.

Biochemical estimation revealed non-significant variation in blood glucose concentration in all three groups of dogs at different time intervals and between the groups and values were within normal range (Table-4). Non-significant change in values of blood glucose with in group and between groups, suggestive of no influence of low level laser therapy on blood glucose. Singh et al. (2011) in dogs and Bhowmick et al. (2013) in calves, also reported non-significant change in blood glucose concentration by treatment of low level laser therapy.

Serum creatinine kinase showed significant increase from 0 day to 7th day in all three groups of dogs, while comparison between three groups of dogs, revealed non-significant difference between three groups (Table-4). Significant increase in values of serum creatinine kinase may be because of release of certain factors during wound healing responsible for causing release of CK in blood. This may also be, to fulfill high energy demands in wounded cells due to faster healing in laser treated wounds, as also observed in present study. Avni et al. (2005) also reported higher content of creatinine phosphokinase activity in LLLT treated muscle wounds as compared to non-irradiated ones.

A significantly high values of total protein in laser treated dogs was observed as compared to control group of dogs. Comparative evaluation between three groups of dogs revealed significantly high values of total protein in laser treated dogs as compared to control group of dogs, although values fluctuated within normal range (Table-5). This may be due more loss of protein in exudate, as more exudaation was observed in group I. However no exudation was observed at 7th day interval in group II and III dogs, due to which there may be no loss of protein in exudate, resulting in increased protein concentration. Siposan and Lukacs (2001) and Bhowmick et al. (2013) also reported significant increase in total proteins in low level laser treated wounds in humans and calves, respectively.

Non-significant increase in serum albumin was observed from 0 day to 7th day interval in all three groups of dogs. Comparison between three groups of dogs, revealed non-significant variation in values of serum albumin (Table-5). Increase in serum albumin may be because of decrease exudation in laser treated wounds, which resulted in to decrease loss of serum albumin in exudate, thereby resulting in increase in serum albumin in blood. Similar findings were also reported by Singh et al. (2011) in dogs and Bhowmick et al. (2013) in calves.

Non-significant variation in values of globulin was observed at different time intervals in all three groups of dogs. However, comparison between three groups of dogs showed significantly high values of globulin in laser treated group II and III, as compared to group I dogs (Table-5). This may be due to decreased exudation in laser treated wounds resulting in decreased loss of total protein in exudates, thereby resulting in increase in amount of globulin in blood.

Plasma fibrinogen showed non-significant decrease from 0-7th day interval in all three groups of dogs. Similarly, comparison between three groups of dogs, revealed non-significant differences between them (Table-5). As formation of clots as well as fibrin network was observed in haematoma due to accumulation of blood, this might have activated fibrinogen synthesis. After operation because of removal of blood clots and synthesis of fibrinogen may have got decreased, resulting in decrease concentration of fibrinogen in blood. Similar findings were also recorded by Kim and Kim (2011) in rats.

Thus on the basis of present study, it was concluded that, low level laser therapy is effective for healing of aural heamatoma, as it decreases inflammation, exudation and embedding of sutures with clinically no scar at wound site after healing, suggestive of anti-inflammatory and cosmetic properties of low level laser therapy. Haematological and biochemical parameters remained within the normal range by laser treatment, suggestive of no adverse effect of laser therapy on vital functions of the body.

References

Anvari, M., Tafti, M.A. and Yazdi, M. (2011). Effect of low level laser therapy on experimental wounds of hard palate mucosa in mice. Indian J. Experiml. Biology 49: 357-61.

Avni, D., Levkovitz, S., Maltz, L. and Oron, U. (2005). Protection of skeletal muscles from ischemic injury: Low level laser therapy increases antioxidant activity. Photomedicine and Laser Surg. 23: 273-77.

Bhowmick, D., Bhargava, M.K., Jaware, S., Shahi, A. and Singh, R. (2013). Efficacy of low level laser therapy on healing of clinical open wounds in calves. Indian J. Vet. Surg. 34: 13-15.

Calin, M.A., Coman, T. and Calin, M.R. (2010). The effect of low level laser therapy on surgical wound healing. Romanian Reports in Physics 62: 617-27.

Hodjati, H., Rakei, S., Johari, H.G., Geramizedeh, B., Sabet, B. and Zeraatian, S. (2014). Low level laser therapy: An experimental design for wound management. J. Cutaneous and Aesthetic Surg. 7: 14-17.

Jann, H.W., Bartels, K., Ritchey, J.W., Payton, M. and Bennett, J.M. (2012). Equine wound healing: influence of low level laser therapy on an equine metacarpal wound healing model. Photonics and Lasers in Med. 1 : 117-22.

Jovanovi, G., Buri, N. and Tijanic, M. (2010). Stimulation of mucoperiostal slice epithelization by small power laser after the primary plastic of oroantral communication. Medicinski Pregled 63: 188-93.

Joyce, J.A. (2000). Canine aural haematoma. Waltham Focus 10: 4-9.

Kim, Y.E. and Kim, E.J. (2011). Anti-inflammation Effect of Low Intensity Laser Therapy in Collagen-induced Arthritis in Rats. Korean J. Oriental Physiol. Pathol. 25: 870-75.

Kumar, A. (2008). Veterinary SurgicalTechniques, 1st Edn., Vikas Publishing Co., New Delhi, p. 208.

Larsen, S. (1968). Intrachondral rupture and haematoma formation in the external ear of dogs. Vet. Pathol. 5: 442-50.

Pallota, R.C., Bjordal, J.M., Frigo, L., Leal Junior, E.C., Tiexeira, S., Marcos, R.L., Ramos, L., Messias, F.de M. and Lopes-Martins, R.A. (2012). Infrared (810-nm) low-level laser therapy on rat experimental knee inflammation. Lasers in Med. Sci. 27 : 71-78.

Rocha, A.M., Vieira, B.J., Andrade, L.C.F. and Aarestrup, F.M. (2009). Low level laser therapy increases transforming growth factor-2 expression and induces apoptosis of epithelial cells during the tissue repair process. Photomedicine and Laser Surg. 27: 303-07.

Singh, M., Bhargava, M.K., Shahi, A., Singh, A.P., Jawre, S., Singh, R., Chandrapuria, V.P. and Kocchar, G. (2011). Efficacy of low level laser therapy on wound healing in dogs. Indian J. Vet. Surg. 32: 103-06.

Siposan, D. and Lukacs, A. (2001). Relative variation to received dose of some erythrocytic and leukocytic indices of human blood as a result of low-level laser radiation: an in vitro study. J. Clin. Laser Med. Surg. 19: 89-103.

Souil, E., Gauthier, B., Sumian, C., Bachelet, M., Buys, B., Polla, B.S. and Mordon, S. (2001). Laser assisted skin closure (LASC) by using a 815-nm diode-laser system accelerates and improves wound healing. Lasers in Surg. Med. 28: 168-75.

Keerti Nema (1), M.K. Bhargava (2), Madhu Swamy (3), Dharmendra Kumar (4) and D. Bhowmick (1)

Department of Veterinary Surgery and Radiology College of Veterinary Science and Animal Husbandry Nanaji Deshmukh Veterinary Science University (NDVSU) Jabalpur - 482001 (Madhya Pradesh)

(1.) Veterinary Assistant Surgeon

(2.) Director (Instruction)/Professor and Corresponding author. E-mail: dr.bhargava@yahoo.co.in

(3.) Professor and Head, Veterinary Pathology

(4.) Assistant Professor
Table -1 : Clinical evaluation at different time intervals

Parameter                Interval    Group I     Group II    Group III
                         (days)

Degree of inflammation      1         ++ to +++   ++ to +++   ++ to +++
                            3         + to ++     - to+       - to+
                            5         - to +      -           -
                            7         -           -           -
                            9         -           -           -
Degree of exudation         1         ++ to +++   ++ to +++   ++ to +++
                            3         ++ to +++   + to ++     +
                            5         + to ++     -           -
                            7         +           -           -
                            9         -           -           -
Degree of embedding         1         -           -           -
of sutures                  3         -           -           -
                            5         + to ++     -           -
                            7         + to ++     +           +
                            9         ++ to +++   +           +

Table-2: Mean values ([+ or -]SE) of haematological parameters

Parameter                 Intervals (days)      Group I

Total erythrocyte count     0                  06.74[+ or -]0.38
(million/[micro]l)             7                  06.81[+ or -]0.20
Total leukocyte count       0                  12.58[+ or -]1.54
(thousand/[micro]l)            7                  14.86[+ or -]0.79
Heamoglobin (g%)            0                  12.28[+ or -]1.72
                            7                  12.75[+ or -]1.47
Packed cell volume (%)      0                  42.53[+ or -]1.05
                            7                  43.78[+ or -]0.45

Parameter                     Group II            Group III

Total erythrocyte count   06.70[+ or -]0.24   06.36[+ or -]0.41
(million/[micro]l)           06.80[+ or -]0.25   06.54[+ or -]0.43
Total leukocyte count     15.25[+ or -]0.28   14.98[+ or -]1.48
(thousand/[micro]l)          16.84[+ or -]0.22   16.57[+ or -]1.25
Heamoglobin (g%)          13.30[+ or -]0.86   13.68[+ or -]1.21
                          14.03[+ or -]1.09   14.45[+ or -]1.29
Packed cell volume (%)    43.35[+ or -]3.40   43.20[+ or -]4.71
                          45.00[+ or -]2.92   44.88[+ or -]4.60

Table-3: Mean values ([+ or -]SE) of differential leukocytes count

Parameter         Intervals (days)     Group I

Neutrophils (%)      0                 68.50[+ or -]1.04
                     7                 70.25[+ or -]0.48A
Lymphocytes (%)      0                 25.00[+ or -]1.08
                     7                 25.00[+ or -]1.00
Monocytes (%)        0                 03.25[+ or -]0.25
                     7                 02.25[+ or -]0.48
Eosinophils (%)      0                 03.25[+ or -]0.25
                     7                 02.50[+ or -]0.29

Parameter            Group II             Group III

Neutrophils (%)   69.25[+ or -]0.85    69.50[+ or -]0.29ab
                  67.75[+ or -]0.75B   66.50[+ or -]0.64cB
Lymphocytes (%)   25.00[+ or -]0.91    24.75[+ or -]0.48
                  26.25[+ or -]0.75    27.50[+ or -]0.65
Monocytes (%)     02.75[+ or -]0.25    02.75[+ or -]0.48
                  03.50[+ or -]0.29    03.50[+ or -]0.50
Eosinophils (%)   03.00[+ or -]0.41    03.00[+ or -]0.41
                  02.50[+ or -]0.29    02.50[+ or -]0.29

Table-4: Mean values ([+ or -]SE) of blood glucose and serum creatinine
kinase

Parameter               Intervals (days)      Group I

Blood glucose (mg/dl)     0                   65.36[+ or -]01.25
                          7                   70.76[+ or -]06.17
Serum creatinine          0                   72.51[+ or -]10.28b
Kinase (U/L)              7                  141.80[+ or -]17.48a

Parameter                    Group II               Group III

Blood glucose (mg/dl)   69.61[+ or -]03.53     69.61[+ or -]00.75
                        71.84[+ or -]03.21     71.86[+ or -]00.71
Serum creatinine        71.76[+ or -]05.52b    73.39[+ or -]03.15b
Kinase (U/L)           134.54[+ or -]32.19a   134.66[+ or -]06.41a

Table-5: Mean values ([+ or -]SE) of total serum protein, serum
albumin, globulin and plasma fibrinogen

Parameter                    Intervals (days)     Group I

Total Serum protein (g/dl)     0                05.53[+ or -]0.29
                               7                04.98[+ or -]0.50A
Serum albumin (g/dl)           0                02.36[+ or -]0.20
                               7                02.57[+ or -]0.24
Globulin (g/dl)                0                03.02[+ or -]0.16
                               7                02.30[+ or -]0.17A
Plasma fibrinogen (g/dl)       0                00.15[+ or -]0.07
                               7                00.11[+ or -]0.16

Parameter                       Group II             Group III

Total Serum protein (g/dl)   05.95[+ or -]0.30    06.47[+ or -]0.32
                             06.20[+ or -]0.23B   06.96[+ or -]0.25B
Serum albumin (g/dl)         02.22[+ or -]0.11    02.27[+ or -]0.04
                             02.68[+ or -]0.17    02.80[+ or -]0.08
Globulin (g/dl)              03.54[+ or -]0.18    03.98[+ or -]0.17
                             03.36[+ or -]0.14B   03.97[+ or -]0.16B
Plasma fibrinogen (g/dl)      0.19[+ or -]0.10     0.22[+ or -]0.15
                              0.16[+ or -]0.10     0.19[+ or -]0.07
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Title Annotation:Research Article
Author:Nema, Keerti; Bhargava, M.K.; Swamy, Madhu; Kumar, Dharmendra; Bhowmick, D.
Publication:Intas Polivet
Article Type:Clinical report
Date:Jul 1, 2016
Words:4093
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