Evaluation of physiological, hematological, and biochemical changes in long bone fracture repair with external skeletal fixators in bovine

Introduction

A fracture is a break in the continuity of hard tissue. Bone heals through regeneration, unlike muscle, skin, and heart tissue, which form scars. The management of long bone fractures remains a significant challenge in large animals. The objective of fracture treatment is anatomic reduction and immobilization of fractured bone by external coaptation methods (Anderson et al., 1994) [3] , internal fixators and external fixator applications (Martens et al., 1998, Vogel and Anderson, 2014) [8,12] . External fixation effectively treats open and closed fractures, limb deformities, non-unions, and arthrodesis (Vogel and Anderson, 2014) [12] . The various changes viz., physiological, hematological and biological changes occurs during the fracture repair in preoperative and post-operative periods. The present study carried out to know the physiological, hematological and biological changes occurs in bovine calf that underwent long bone fracture repair by various external skeletal fixators.

Materials and Methods

The 18 bovine calves with long bone fracture were selected for the present study and divided in three groups. The each groups having six cases which treated with linear external skeletal fixation (group-I), circular external skeletal fixation (group-II) and free form external skeletal fixation (group-III). The physiological parameters assessed preoperatively and post operatively. Blood samples were obtained from the jugular vein using K 3 EDTA vials both before and after the surgery at intervals of 15 th , 30 th , and 45 th days and various parameters including hemoglobin (gm/dL), packed cell volume (%), total leukocyte count (10 3 cells/cumm), total erythrocyte count (10 6 cells/cumm), and the components of the Differential Leukocyte Count (DLC) such as neutrophils (%), lymphocytes (%), eosinophils (%), and monocytes (%) were studied. A 7ml blood sample was drawn from the jugular vein into a clot activator tube. The serum was then separated, and biochemical parameters including serum calcium (mg/dL), serum phosphorus (mg/dL), and serum alkaline phosphatase (IU/L) were assessed both pre-and post-operatively on the 15 th , 30 th , and 45 th days.

Results and Discussion

Physiological Parameters

In Group-I, mean of respiration rate per minutes was 21.00±1.24 pre operatively while post-operative it was 24.17±2.59 and in Group-II, mean of respiration rate per minutes was 27.17±1.85 pre operatively while post-operative it was 27.83±1.80,whereas,in Group-III, mean of respiration rate per minutes was 24.67±3.82 pre operatively while postoperative it was 20.17±1.42.The current study found no statistically significant alteration (P > 0.05) within groups during both the pre-operative and post-operative periods. While, in between groups significant (P < 0.05) difference was observed post-operatively in group-II and III whereas, no significant (P > 0.05) difference observed in group-I. This might be due to excitement during the recovery period from general anesthesia in calves. Similar to the study, Yadav et al. (2021a) [13] found a non-significant increase in respiratory rates on the first post-operative day in all animals, followed by a decrease from the third day in bovines treated for tibial fractures The mean of heart rate per minutes was 77.83±8.39 pre operatively while post-operative it was 75.17±9.20 in Group-I. In Group-II, mean of heart rate per minutes was 68.33±3.74 pre operatively while post-operative it was 68.00±4.48, whereas Group-III, mean of heart rate per minutes was 71.33±4.90pre operatively while post-operative it was 67.00±6.81. The present study found no noteworthy (P > 0.05) alteration in mean value of heart rate within the groups or between the groups at pre and post-operative period. The present findings were in accordance with Das et al. (2020) [6] and Ajjodi et al. (2022) [2] . The mean value of rectal temperature in 0 F was 101.33±0.71pre operatively while post-operative it was 100.63±0.44 in group-I. In Group-II, mean value of rectal temperature in 0 F was 101.70±0.52 pre operatively while post-operative it was 100.63±0.57, whereas in Group-III, mean value of rectal temperature in 0 F was 101.57±0.53 pre operatively while post-operative it was 100.33±0.21. The present study found no substantial (P > 0.05) difference in mean value of rectal temperature within the groups or between the groups at pre and post-operative periods. These might be due to administration of a non-steroidal antiinflammatory drug at the time of preoperative preparation which reduce the temperature. The similar findings were observed by Prasad (2013) [10] and Das et al. (2020) [6] .

Hematological Parameters

There were no significant difference observed in mean value of haemoglobin, packed cell volume, total erythrocytes count, and total leucocyte count on day 0, 15 th 30 th and 45 th post days, but in differential leucocytes count significant differences was observed in neutrophils and lymphocytes. The mean value of neutrophils on 0 day was 47.17±0.75 and on 15 th , 30 th and 45 th post-operative days the values were 43.50±0.62, 40.83±0.17and 38.83±0.48, respectively in group-I. The mean value of neutrophils on 0 day was 47.50±1.18 and on 15 th , 30 th and 45 th post-operative days the values were 43.17±1.14, 40.50±0.62 and 37.83±0.75, respectively in group-II, whereas, mean value of neutrophils on 0 day was 48.67±0.76 and on 15 th , 30 th and 45 th post-operative days the values were 43.33±0.42, 40.67±0.56 and 36.83±0.75, respectively in group-III. In comparison between groups, there was no substantial (P > 0.05) differences in mean value of neutrophils at different time interval. While, in within the groups, mean value of neutrophils was significantly (P < 0.05) higher at 0 day as compared to 15 th , 30 th and 45 th post-operative days in group-I and III. However, the mean value of neutrophils was significantly (P < 0.05) lower at 15 th , 30 th and 45 th post-operative days in group-II as compared to 0 day. The higher the neutrophils count at the time of presentation of case might be due to extent of trauma that may leads the inflammation and decreasing counts indicating a reduction in inflammatory response allowing progressive fracture healing. These findings were in concurrence with the findings of Yadav et al. (2021a) [13] and Sravanti et al. (2022) [11] . On the 0 day, mean value of lymphocytes was 48.17±0.70 whereas, on the 15 th , 30 th and 45 th post-operative days values were 51.83±0.48, 56.33±0.49 and 58.50±0.56, respectively in group-I. On the 0 day, mean value of lymphocytes was 47.83±1.08 whereas, on the 15 th , 30 th and 45 th post-operative days values were 51.67±0.88, 55.33±0.8 and 58.17±0.83, respectively in group-II. The mean value of lymphocytes was 46.83±0.79 on day 0 whereas, on the 15 th , 30 th and 45 th postoperative days values were 51.67±0.42, 55.50±0.76 and 59.83±0.60, respectively in group-III. In present study, no significant (P > 0.05) differences was found in mean value of lymphocytes between the groups at different time interval. The mean lymphocytes in within the groups was significantly (P < 0.05) higher at 15 th , 30 th and 45 th post-operative days as compared to 0 days in group-I and III. While, nonsignificantly (P < 0.05) higher at 15 th and 30 th day and significantly (P < 0.05) higher at 45 th post-operative days as compared to 0 day in group-II. This could be attributed to the fluctuation in neutrophil count, which initially increases after surgical intervention, prompting the production of immuneregulatory cytokines by macrophages. These cytokines are involved in activating adrenal axes and stimulating the production of glucocorticoids, which may result in the breakdown of lymphoid tissue and a rise in circulating lymphocytes. These findings were in accordance with those observed by Avasthi (2012) [5] and Sravanti et al. (2022) [11] . The no significant difference was observed in mean value of monocytes and eosinophils.

Biochemical Parameters Serum Calcium

In group-I, the average serum calcium level on day 0 was 8.67±0.11, while on the 15 th , 30 th , and 45 th postoperative days, the values were 9.38±0.32, 9.17±0.18, and 8.98±0.15, respectively. In group-II, the mean serum calcium level on day 0 was 8.62±0.25, and on the 15 th , 30 th , and 45 th postoperative days, the values were 9.38±0.30, 9.07±0.18, and 8.72±0.24, respectively. In group-III, the average serum calcium on day 0 was 9.72±0.55whereas, on the 15 th , 30 th and 45 th post-operative days values were 10.32±0.55, 9.47±0.24 and 9.4±0.30, respectively. The current study found no significant (P > 0.05) differences in the mean serum calcium between the groups at different time intervals. The comparisons within the groups found level of serum calcium was non-significantly (P > 0.05) increased at 15 th , 30 th and 45 th days as compared to 0 day in all the groups. The decreasing the serum calcium level on 30 th post-operative days might be due to excessive calcium deposition at fracture site and increasing the level on 45 th day as comparison to 0 day it may be due to remodeling phase. These findings were in concurrence with the findings Kumar et al. (2018) [7] and Prajapati (2021) [9] .

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