Haemato-biochemical variations during mandibular fracture with bone defect fixation in dogs using deproteinized microcrystalline hydroxyapatite complex granules along with titanium miniplates

Introduction

Fracture is a break in the continuity of bone or hard tissue (like cartilage). It can be partial or complete, and may involve one or multiple bones. Occurrence of mandibular fracture is 10%, in that miniature, less than one year old and males are most commonly involved (Minar et al. 2013) [6] . Titanium mini-plates were successfully used to repair maxillary and mandibular fractures in dogs and cats, achieving rigid fixation, maximum immobilisation, and early return to eating within six to nine weeks (Bilgili and Kurum, 2003) [2] . Hydroxyapatite, being chemically identical to the inorganic matrix of natural bone, offers synthetic processability, favorable biocompatibility, and osteoconductive properties suitable for filling bone voids and defects, with minimal resorption and rapid osseous integration (Mamun et al. 2009) [5] . Preoperative physiological and haemato-biochemical evaluations are vital for assessing surgical fitness, while post-operative monitoring aids in understanding fracture healing and complications. This study examined changes in these parameters during mandibular fracture repair in six dogs using deproteinized microcrystalline hydroxyapatite complex granules along with Titanium miniplates.

Materials and Methods

The present study was carried out among the six clinical cases of dogs presented to the Department of Veterinary Surgery and Radiology, Veterinary College Hospital, Hebbal, Bengaluru. Mandibular fracture with bone defect cases were selected for the study with irrespective of age, breed and sex. For hematological and biochemical assessment, blood samples were obtained from the cephalic or saphenous veins of all six dogs. Hematological parameters-including Total Erythrocyte Count (TEC, 10 6 cells/cmm), Total Leukocyte Count (TLC, 10 3 cells/cmm), Hemoglobin (Hb, g/dL), and Differential Leukocyte Count (DLC,%)-were analyzed using the Mindray BC-2800 Vet automated hematology analyzer. Biochemical indices such as serum calcium (mg/dL), serum phosphorus (mg/dL), alanine aminotransferase (IU/L), alkaline phosphatase (IU/L), and serum creatinine (mg/dL) were evaluated using micro lab instruments RX-50 Biochemical semi-auto analyzer using diluent reagents as per the manufacturer guidelines. The resulting hematological and biochemical data were subjected to statistical evaluation through oneway Analysis of Variance (ANOVA) with the computerbased statistical program GraphPad Prism.

Results and Discussion

3.1 Haematological analysis 3.1.1 Total erythrocyte count (10 6 /cmm) The pre-operative Mean ± SE of total erythrocyte count was 5.67 ± 0.34 and post-operatively varied from 5.50 ± 0.43 to 6.51 ± 0.27. The values were gradually increased from 7 th post-operative day to 60 th post-operative day, this might be due to normal erythropoiesis. All recorded values were within normal range and statistically non-significant (p > 0.05). These findings were in agreement with Srinivasa Murthy (2015) [13] , and Reddy et al. (2020) [11] .

Haemoglobin (gm/dL)

The pre-operative Mean ± SE of haemoglobin level was 12.55 ± 0.86 gm/dL. In post-operative days ranged from 12.03 ± 0.83 to 12.54 ± 0.63. From post-operative day 7 to day 60 haemoglobin values gradually increased, yet remained within the normal expected range and statistically non-significant (p > 0.05). This subtle upward trend might be attributed to natural erythropoietic activity. These findings were in agreement with the findings of Patil et al. (2017) [7] and Reddy et al. (2020) [11] .

Total leucocyte count (10 3 /cmm)

The pre-operative Mean ± SE of total leucocyte count was 15.33 ± 1.88. Post-operatively varied from 12.86 ± 0.80 to 16.15 ± 1.96. During the early post-operative days slight increase in the total leucocyte count was noticed and thereafter values were reduced gradually up to 60 th day. This transient leucocytosis might be attributed to corticosteroid release influenced by factors such as pain, anaesthesia, trauma, surgical intervention, and localized inflammation. All obtained values were within the normal range and statistically non-significant (p > 0.05). Similar observations were documented by Mahesh (2009) [4] , and Reddy et al. (2020) [11] . Rashmitha (2022) [10] found a statistically significant difference in the total leucocyte count in their study on mandibular plating in dogs.

Differential leucocyte count

Mean ± SE value of neutrophils (%), lymphocytes (%), monocytes (%) and eosinophils (%) counts post-operatively varied from 70.36 ± 3.08 to 74.32 ± 1.85, 18.85 ± 1.82 to 21.90 ± 2.38, 4.10 ± 0.61 to 5.08 ± 0.03, and 2.73 ± 0.30 to 3.10 ± 0.24. During the study period all observed values were within the normal range and statistically nonsignificant (p > 0.05). The neutrophil count was higher on initial post-operative days and thereafter reduced gradually. This could be attributed to a diminished inflammatory response, facilitating continuous fracture healing in the absence of exudate formation. Similar findings were reported by Madhura (2013) [3] , and Srinivasa Murthy (2015) [13] .

The pre-operatively noticed Mean ± SE value of the serum creatinine was 0.88 ± 0.07. Post-operatively up to 60 th day, ranged from 0.77 ± 0.06 to 0.97 ± 0.06. Throughout the study period, the values were fell within the normal range and statistically non-significant (p > 0.05). The findings indicate that the anaesthetic protocol, antibiotic and analgesic administration, and MCHC granules utilized for this study had no adverse impact on renal function. These findings were in accordance with Rashmitha (2022) [10] and Yelmule et al. (2025) [15] .

Serum Alanine aminotransferase (IU/L)

The Mean ± SE value of pre-operative serum alanine aminotransferase was 43.53 ± 5.70. Post-operatively up to 60 th day, Mean ± SE of serum alanine aminotransferase level ranged from 33.14 ± 2.62 to 45.62 ± 2.32. All observed values were within the normal range during the study period and statistically non-significant (p > 0.05). Similar findings were reported by Pooja et al. (2023) [9] and Yelmule et al. (2025) [15] .

Serum Alkaline phosphatase (IU/L)

The Mean ± SE value of pre-operative serum alkaline phosphatase was 127.90 ± 12.19. Post-operatively up to 60 th day, Mean ± SE of serum alkaline phosphatase level ranged from 132.20 ± 7.07 to 160.94 ± 11.84. The mean serum alkaline phosphatase (ALP) levels exhibited a gradual rise up to the 30 th post-operative day, then gradually decreased up to 60 th day. Although the increase in ALP was statistically non-significant (p > 0.05), it likely reflects enhanced chondroblastic activity contributing to bone regeneration. The findings were in concurrence with Rashmitha (2022) [10] and Yelmule et al. (2025) [15] . Whereas Phaneendra et al. (2016) [8] found a significantly increase in the serum alkaline phosphatase values up to 14 th postoperative day and values were reached the normal range by 21 th post-operative day.

The Mean ± SE value of pre-operative serum calcium was 9.82 ± 0.17. Post-operatively values were varied from 9.41 ± 0.32 to 10.48 ± 0.34. The decrease of serum calcium was noticed from pre-operative day to post-operative 7 th day. Thereafter it gradually increased, with a peak was noted on 30 th day. The initially reduced serum calcium concentration was likely due to its increased utilization at the fracture site, where it was deposited as calcium salts. In later postoperative days serum calcium values were gradually increased within normal limits and statistically nonsignificant (p > 0.05) might reflect hormonal regulation of calcium metabolism. The observed results were in agreement with Yashvanth (2023) [14] and Yelmule et al. (2025) [15] . On the contrary, Srinivas (2022) [12] observed that serum calcium levels were increased up to 15 th postoperative day and thereafter decreased gradually up to 60 th day in their study.

The pre-operative Mean ± SE value of serum phosphorus was 4.07 ± 0.47. It varied from 3.96 ± 0.27 to 4.35 ± 0.26 post-operatively up to 60 th day. The gradual increase in mean serum phosphorus levels occurred up to 7 th post-operative day, thereafter decreased gradually. Statistically non-significant (p > 0.05) fluctuations were noted at various points and all values remained within expected normal range. The increase in the serum phosphorus levels indicated the formation of new bone during the mineralisation of extra cellular matrix. The progressive decline in serum phosphorus concentration might be attributed to osteoclastic activity at the fracture site, resulting in the resorption of necrotic bone tissue. The observed results were in agreement with Pooja et al. (2023) [9] and Yelmule et al. (2025) [15] . However, Bidari et al. (2023) [1] in their study reported that serum phosphorous level was significantly increased on 30 th post-operative day thereafter it decreased by 60 th post-operative day.

Conclusion

The haematological and biochemical evaluations revealed that all parameters remained within normal physiological limits throughout the study period, with only minor, nonsignificant fluctuations. Transient leucocytosis and elevated neutrophil counts in the early days indicated short-lived inflammatory responses that subsided with healing. Biochemical markers such as serum creatinine and alanine aminotransferase confirmed that anaesthetic, antibiotic, analgesic protocols, and MCHC granules did not adversely affect renal or hepatic function. Serum alkaline phosphatase exhibited a non-significant rise, suggestive of chondroblastic activity and bone regeneration during fracture healing. Serum calcium and phosphorus levels showed minor fluctuations, reflecting their utilization in mineral deposition and remodeling at the fracture site. Overall, haematological and biochemical findings demonstrated stable systemic health, with observed trends supporting effective fracture healing and biocompatibility of MCHC granules.

Acknowledgement

We are thankful to Department of Veterinary Clinical Complex, Veterinary College, Hebbal, Bengaluru for the facilities provided and Fredun Pharmaceutical Ltd., Mumbai, for providing FREOSSI® granules for this research work.

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