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ThesisItem Open Access Osteosynthesis in induced tibial segmental defect using co-precipitated hydroxyapatite gel and titanium mesh cage in rabbits (Oryctolagus cuniculus)(G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-09) Karki, Nikita; Karki, Nikita; Das, Arup Kumar; Das, Arup KumarThe present study was conducted in (12) clinically healthy adult New Zealand white rabbits (1.5-2.0 years old), of either sex weighing 1.5-2.0 kg and they were subjected to induction of segmental defect in tibia under general anaesthesia. Thereafter, insertion of intramedullary titanium pin along with cylindrical titanium mesh cage and injectable co-precipitated HA gel used for reconstruction of segmental defect in tibia. All rabbits were divided randomly in two groups (A and B) with 6 animals in each group. In rabbits of Group A fracture fragments were stabilized with titanium mesh and titanium screws. While in rabbits of Group B, fracture fragments were stabilized with titanium mesh cage, reinforced with intramedullary pin and injectable co- precipitated HA ladened Mg-gelatin-collagen gel was used. The common features included in each rabbits of both the groups were anaesthetic regimen – premedication with glycopyrrolate, xylazine and induction of ketamine anaesthesia was pursued. Maintenance of anaesthesia was done with Isoflurane @ 1-1.5 percent concentration of with 100 percent oxygen saturation. On the basis of evaluation of parameters, better wound healing was observed in animals of Group B as compared to animals of Group A. On the basis of biochemical parameters, faster and better wound healing was observed in rabbits of Group B. Estimation of haematological and biochemical parameters were done at day 0, 7, 15, 30 and 60. There was a significant increase in serum values of ALP, calcium, phosphorus, blood glucose and decrease in magnesium, total protein. Radiological findings showed faster rate of healing and earlier reduction in gap (1.00 cm) and moderately higher radio-denser area at induced segmental defect in tibia of animals of Group B. On gross observation of test bones, negligible gap was observed at induced segmental defect in animals of Group B depicting faster healing, mineralization of developed callus and bridging of segmental defect as compared to animals of Group A. There was new bone formation found around the external circumference of the cage which was bridging the proximal and distal bone cage junction. Exuberant callus formation was found around the cage in animals of Group B. The implant remained completely stable, in correct alignment and found in pre-experimental state in animals of Group B. Titanium mesh cage used here act as scaffold for bony restoration, allows vascular supply both to the mucosa and bone during regeneration phase. The construct used here for osteosynthesis of induced segmental defect at tibia provided adequate mechanical strength, augment limb stability and early bone continuity. Conclusively a better fracture healing was observed in rabbits of Group B as compared to rabbits of Group A and it reveals that the co-precipitated HA gel used act as scaffold for bony restoration and hence it possess attributes of biodegradability, biocompatibility. The above mentioned technique can be use for treatment of long bone segmental defect as it has got wide scope to be used in clinical veterinary practice. In the present study the outcome of surgical management of large segmental defects in tibia using cylindrical titanium mesh cage was found to be effective for treatment of LBSD.ThesisItem Open Access Evaluation of healing efficacy of topical herbal formulations on experimental goat model(G. B. Pant University of Agriculture and Technology, Pantnagar, 2022-09) Ghalaut, Bharat; Das, Arup KumarThe present study was conducted on six male goats. Six full thickness incisional wound of 1 inch length and six excisional wounds of 1 inch × 1 inch dimension were created on both side of the back with an objective to evaluate the healing efficacy of four different topical polyherbal formulations. One positive control was treated with Soframycin ointment from day of wound creation till recovery. The other four test sites were treated with Charmil Plus gel, Charmil Spray, AV/CPS/23 lotion and Brand A respectively. One wound site was left untreated as negative control. The wounds were evaluated on the basis of clinical, macroscopical and haematobiochemical studies. Clinical and haematobiochemical parameters did not reveal any significant change between the different treatments and were variable within normal physiological limits. For clinical evaluation- quantity of exudation, wound contraction and rate of wound healing was observed. The result revealed less exudation with better healing rate and wound contraction in test product Brand A followed by Charmil spray, AV/CPS/23, and Charmil ointment in decreasing order; and all the wounds that got treatment were found to have better healing than the negative control woundThesisItem Open Access Studies on the effects of tiletamine-zolazepam, propofol and sevoflurane anaesthesia in dexmedetomidine and butorphanol premedicated dogs(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-07) Tiwari, Kanika; Jadon, Narendra SinghThirty-six adult dogs used in the study were randomly divided into 6 subgroups (A-1, A-2, A-3, B-1, B-2 and B-3). All the dogs were administered with atropine sulphate at the dose rate of 0.04 mg/kg body weight subcutaneously. After five minutes, animals of group A-2 and B-2 were premedicated with dexmedetomidine whereas animals of group A-3 and B-3 were premedicated with dexmedetomidine and butorphanol at the dose rate of 5 μg/kg body weight and 0.1 mg/kg body weight intravenously. The animals of groups A-1, A-2, A-3, B-1, B-2 and B-3 induced with tiletamine-zolazepam and propofol at the dose rate of 6.66±0.06 mg/kg iv, 1.86±0.11 mg/kg iv, 1.26±0.02 mg/kg iv, 5±0.06 mg/kg iv, 2.56±0.01 mg/kg iv and 2.16±0.01 mg/kg iv respectively. Anaesthesia was maintained with sevoflurane and MAC of sevoflurane was determined by tail clamp method in all the animals. The level of anaesthesia was determined by observing various clinical (induction time, duration of anaesthesia, muscle relaxation, pedal reflex, palpebral reflex, analgesia, recovery time, sternal recumbency time, standing time, complete recovery time, required doses of induction agents and reduction in the % MAC of sevoflurane, physiological and haemodynamic (rectal temperature, heart rate, respiration rate, blood pressure, haemoglobin oxygen saturation (SpO2), electrocardiography), haematological (haemoglobin, total erythrocyte count, total leucocyte count, differential leucocyte count) and biochemical (serum glucose, total protein, serum albumin, serum urea nitrogen, serum creatinine, alanine amino transferase, aspartate amino transferase) parameters. Clinical efficacy of the anaesthetic combination was determined by performing various surgical operations.The mean induction time reduced in the animals of groups A-2, A-3, B-2 and B-3 after the administration of preanaesthetic drugs whereas longer duration of anaesthesia, recovery time, sterna recumbency time, standing time and complete recovery time were observed in the animals of group A-2, A-3, B-2 and B-3 after the administration of preanaesthetic drugs. Adequate muscle relaxation and excellent analgesia was observed in the post-induction period in all the animals of different groups. Pedal and palpebral reflexes were very weak after the induction with tiletamine-zolazepam in group A-1, A-2 and A-3 whereas it was completely lost after the induction with propofol in group B-1, B-2 and B-3. Preanaesthetic administration of dexmedetomidine and butorphanol reduced the induction dose of drugs (propofol and tiletamine-zolazepam) and MAC of sevoflurane. The rectal temperature, heart rate and respiration rate showed significant decrease at various time interval and reached to the baseline values by 24 hours post-anaesthesia. SAP, DAP and MAP was decreased non-significantly in group A-1 and B-1 whereas it showed a significant increase in group A-2, A-3, B-2 and B-3 at respective time interval. Significant decrease in haemoglobin oxygen saturation was observed after administration of induction agents. Significant reduction in PR interval was observed in animals of group A-1 whereas non-significant changes were observed in QRS and QT interval in all the groups of animals. Inverted T-wave and sinus dysrhythmia were observed after administration of dexmedetomidine whereas sinus tachycardia was observed after induction with tiletamine-zolazepam. Transient changes within physiological limits were observed in haemato-biochemical parameters which reached to baseline values by 24 hours. On the basis of above mentioned parameters, it was observed that the anaesthetic combinations used in the study have least deleterious effects on different body systems and the most efficacious anaesthetic combination was atropine sulphate-dexmedetomidine-butorphanol-tiletamine-zolazepam-sevofluraneThesisItem Open Access Anaesthetic evaluation of various combinations of xylazine, ketamine and tiletaminezolazepam in dogs(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-07) Sulekha; Kandpal, ManjulEighteen dogs of either sex, different breed, body weight and age (requiring various surgical corrections) were randomly divided into three groups namely A, B and C. Atropine sulphate (0.025mg/kg SC) was administered in all the groups followed by tiletamine-zolazepam (till effect IV) in group A, xylazine (0.5mg/kg IM) and tiletamine-zolazepam (till effect IV) in group B and xylazine (0.5mg/kg), ketamine (2.5mg/kg) and tiletamine-zolazepam (till effect IV) in group C. The induction dose of tiletamine-zolazepam observed in group A, B and C were 6.5, 5.5 and 4 mg/kg body weight respectively on intravenous administration. The efficacy of the anaesthesia was evaluated by observing clinical (induction dose, induction time, duration of anaesthesia, sternal recumbency time, complete recovery time, jaw relaxation score, pedal reflex score and palpebral reflex score), physiological (heart rate, respiration rate, rectal temperature, capillary refill time, systolic arterial pressure, diastolic arterial pressure, mean arterial pressure and haemoglobin oxygen saturation), haematological (haemoglobin, total erythrocyte count, packed cell volume, total leucocyte count and differential leucocyte count) and biochemical (total protein, plasma glucose, blood urea nitrogen, serum creatinine, total bilirubin, alanine aminotransferase and aspartate aminotransferase) parameters before and after administration of anaesthetic drugs. Anaesthetic effects like analgesia and muscle relaxation was observed to be slightly better in group C in comparison to group A and B. Heart rate increased and respiration rate decreased post induction in all three groups. Haematological parameters like haemoglobin, total erythrocyte count, packed cell volume, total leucocyte count, lymphocytes, monocytes, eosinophils and basophils showed transient decrease whereas neutrophils showed transient increase in all the groups at 30 and 60 minutes post induction and approximately approached baseline value by 24 hours. Biochemical parameter like total protein showed transitory decrease whereas plasma glucose, blood urea nitrogen, serum creatinine, total bilirubin, alanine aminotransferase and aspartate aminotransferase reported an increase in all the groups at 30 and 60 minutes post induction and approached baseline values by 24 hours. These changes in haematological and biochemical parameters were within normal physiologic range. Tiletaminezolazepam with its fast and smooth induction, intermediate duration of action, excellent muscle relaxation and good compatibility with xylazine and ketamine, was found to be an effective general anaesthetic either alone or in combination. Group C with xylazine, ketamine and tiletamine-zolazepam combination required significantly lower induction dose and provided longer duration of anaesthesia in comparison to group A and group B.ThesisItem Open Access Studies on echostructural pattern of urinary system in adult female goats(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2022-07) Singh, Shivani; Das, Arup KumarB-mode ultrasonographic study of urinary system was performed in 20 healthy adult female Jamunapari goats. Mean±S.E. values of body weight and age of selected animals were 27.49±0.79kg and 2.77+0.10 years, respectively. All twenty animals were subjected to sonographic examination after clipping the hairs from desired sites to scan the various organs of urinary system in standing position under physical restraint without using any anesthetic agent. The values of various clinico-physiological parameters including rectal temperature, heart rate, respiration rate, colour of mucous membranes were recorded to examine the health status prior to sonography. The left and right kidneys were scanned from right dorsal flank by paravertebral approach using a transcutaneous micro-convex probe at 5 MHz and a convex probe at 2.8-6 MHz frequency. A dual frequency (5-9 MHz) micro-curved array cavity probe was used to scan bladder and urethra per rectally. The renal parenchyma in healthy goats was distinctly divided into hypoechoic outer cortex and anechoic inner medulla. The hyperechoic renal sinus complex (fat, vessels, lymphatics and fibrous tissue) was present in centre of the kidney and renal pelvis was anechoic. The renal capsule was seen as hyperechoic fine line surrounding the renal parenchyma. Pulsed wave renal color doppler ultrasound was performed in 3 goats out of the 20 with the aim to generate reference images of renal vasculature in healthy goats. The mean±S.E. values of renal length, renal height, horizontal diameter of renal sinus, vertical diameter of renal sinus and diameter of renal pelvis for right kidney were 6.11±0.09, 3.26±0.08, 3.01±0.11, 1.32±0.03, 1.05±0.03, respectively while for left kidney these values were 6.02±0.09 and 3.35±0.07, 2.67±0.12, 1.14±0.06, 1.10±0.07 cm. The ureters could not be visualized in any of the goats. The distended bladder was seen as anechoic circular structure surrounded by a smooth echogenic wall with a mean thickness of 1.65+0.06 mm. No significant difference was observed in the measurements of both kidneys except in dimensions of renal sinus. Moderately positive correlation was found between renal size and body weight of goats. It was concluded that ultrasound is an easy, rapid and non-invasive method to evaluate echotexture and morphometry of structures of urinary system.ThesisItem Open Access Comparative evaluation of anaesthetic effects of various combinations of acepromazine, butorphanol, dexmedetomidine and ketamine in glycopyrrolate premedicated dogs maintained with Sevoflurane(G.B. Pant University of Agriculture and Technology, Pantnagar, District Udham Singh Nagar, Uttarakhand. PIN - 263145, 2021-02) Kundu, Saikat Kumar; Kandpal, ManjulEighteen adult dogs (requiring various clinical procedures) used in this study were divided randomly into three subgroups (A, B and C). All the dogs were subjected to the administration glycopyrrolate at the dose rate of 0.01 mg/kg intramuscularly followed by acepromazine given as pre anaesthetic at the dose rate of 0.04 mg/kg IM in group A, butorphanol at the dose rate of 0.4 mg/kg IV in group B and dexmedetomidine 10 μg/kg IV in group C. Ten minutes after premedication, all the animals of each group were induced with intravenous ketamine. The induction doses were found to be 7.62±0.09 mg/kg in group A, 7.12±0.09 mg/kg in group B and 6.69±0.08 mg/kg in the animals of group C respectively. Intubation was easy and free of any major complications in the animals of group A and group C, however in the animals of group B, they showed strong coughing reflex, chewing motion and strong tongue with-drawl during the time of intubation. Anaesthesia was maintained in all the groups of animals with sevoflurane. The minimum concentration of inhaled sevoflurane for maintenance of anaesthesia was noted in all the animals by the tail clamp method. The level of anaesthesia was determined by observing various clinical (Induction time, intubation score, duration of anaesthesia, muscle relaxation, pedal reflex, recovery time, complete recovery time, recovery score, required doses of different drugs and minimum concentration of inhaled sevoflurane, physiological (rectal temperature, heart rate, respiration rate, blood pressure, capillary refill time, haemoglobin oxygen saturation-SpO2, electrocardiography), haematological (haemoglobin, total erythrocyte count, erythrocyte sedimentation rate, total leucocyte count, differential leucocyte count, packed cell volume, and biochemical (serum glucose, total protein, serum urea nitrogen, serum creatinine, alanine amino transferase ,aspartate amino transferase and total bilirubin) parameters. The animals of group C showed quicker induction. All anaesthetic combinations tested in the study were able to provide adequate muscle relaxation but group C showed better result. The pedal reflex was totally lost after administration of the induction agents in all the groups. The recovery time, complete recovery time were highest in group B followed by group C and group A. The minimum concentration of inhaled sevoflurane for maintenance of anaesthesia were found to be (1.68 ± 0.47)% for group A, (2.00 ± 0.25)% for group B and (1.85 ± 0.54)% for group C. The rectal temperature, heart rate and respiration rate showed significant decrease at various time intervals. The systolic, diastolic and mean arterial pressures showed significant decrease in its values in the groups A but not in the group B and group C, and returned to their base values by 90 minutes post anaesthesia. In all the groups of animals the capillary refill time (CRT) and haemoglobin oxygen saturation (SpO2) were always less than 2 seconds and more than 95% respectively. The haemoglobin, PCV, TLC and TEC values of the animals in various groups showed significant decrease in its values whereas the ESR showed a significant increase. Significant changes were observed in differential leucocyte count. Serum glucose, Serum urea nitrogen, creatinine, ALT, AST and total bilirubin levels increased significantly in all the groups of animals however there was significant decrease in total protein levels. On the basis of above mentioned parameters, it was concluded that the most efficacious anaesthetic combination was glycopyrrolate (0.01 mg/kg IM) dexmedetomidine (10 μg/kg IV)- ketamine (6.69±0.08 mg/kg IV) and sevoflurane (1.85 ± 0.54) % in the animals of group C and it was found suitable for surgeries because of its smooth induction, less induction time, excellent muscle relaxation and analgesia properties, maximum dose sparing effect on ketamine and smooth and uneventful recovery.ThesisItem Open Access Studies on applicability of titanium mesh cage for osteosynthesis of induced long bone segmental defects in rabbits (Oryctolagus cuniculus )(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-08) Raj Kumari; Das, Arup KumarThe present study was conducted in two phases. In the first phase suitable anaesthetic protocol, for the development of orthopaedic model in rabbits, was explored. It was conducted in twenty (20) clinically healthy adult New Zealand white rabbits (~1.5-2.0 years old), of either sex, weighing 1.5 to 2.0 kg and were arbitrarily allocated for anaesthetic protocol-1 and anaesthetic protocol-2 containing 10 rabbits in each protocol. The common features included in these two anaesthetic protocols that premedication and induction agents along with dose rate were kept similar and an interval of five minutes was kept between each of these injections. Afterwards anaesthesia was maintained by sevoflurane in rabbits of protocol-1 and with isoflurane in rabbits of protocol-2. On the basis of evaluation of anaesthesia related parameters - sevoflurane was found relatively a better maintenance anaesthetic agent. Thereafter sevoflorane was used during the model development and surgical repair of left tibial midshaft segmental defects (LBSDs). In the second phase of study sixteen (16) adult New Zealand white rabbits (~1.5-2.0 years old) of either sex, weighing 1.5 to 2.5 kg were selected and were divided randomly into two groups (A and B) with eight animals in each group. In these rabbits LBSDs were created and cylindrical Ti-mesh cages and full length intramedullary Ti K-wire were used as implants for osteosynthesis. In rabbits of Group-B Ti-mesh cages were filled with HA ladened Mg-gelatin-collagen-spacer. On the basis of evaluation of parameters, better wound healing was observed in rabbits of Group-B compared to Group-A rabbits. Evaluation of biochemical parameters also showed greater intensity in healing of LBSDs of Group-B. The radiological parameters showed faster rate of new bone formation, reduction in gap (1.00 cm) and remodeling of segmental defect in rabbits of Group- B. On gross observations of segmental defect at the test limb it was found that a clear visible new bone bridging the entire defect and was closely integrated with the proximal and distal host bone-cage junction was observed in Group-B rabbits. However a slightly visible gap was observed in Group -A animals of this study. Although in both the groups the implant materials remained completely stable and in correct alignment that might have adequately aided in process fracture healing. The findings of histopathological and tetracycline labeling were also corroborating the findings of clinical and radiological parameters. Conclusively a better fracture healing and bridging of critical size defect was observed in rabbits of Group B as compared to Group -A rabbits and it can be stated that spacer used in this group might have aided for it and possessing attributes of osteogenicity, biodegradibity and biocompatability. In summated form it can be stated that the outcome of surgical management of large segmental defects of the tibia using cylindrical titanium mesh cages has advantages of comparative simplicity and versatility. So it has the scope to be used for the treatment of long segmental bone defects in clinical veterinary practice. However, further clinical studies are required in order to evaluate of its efficacy.ThesisItem Open Access Clinical and radiographic study on surgical management of distal diaphyseal fracture in long bones by end threaded positive profile intramedullary pins and titanium elastic nails in dogs(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-08) Manish Kumar; Kandpal, ManjulThe present study was conducted on 12 clinical cases of dogs of either sex with distal diaphyseal long bone fractures. The animals were randomly divided into two groups having equal numbers viz. Group A in which the fracture was stabilized with End threaded positive profile intramedullary pins and Group B by Titanium elastic nails. Diagnosis of the distal diaphyseal fracture done by preoperative clinical evaluation and radiography. Various parameters like clinic-physiological parameters like rectal temperature, heart rate, respiration rate, inflammation, pain score, lameness and weight bearing score, Haematobiochemical parameters like Hb, TEC, TLC,DLC & ALP were recorded in all the animals at various time interval. Clinico-physiological parameters did not show any significant difference between the both groups and were fluctuating within normal physiological range. Intra operative observation like duration of surgery, radiation exposure time, extent of manipulation and soft tissue damage, degree of technical difficulties, status of fracture reduction and fracture fixation was also recorded. The mean duration of surgery and intraoperative radiation exposure time significantly less in Group A compare to Group B. Lameness score was significantly less in group B compare to group A. The weight bearing score was significantly more in Group B as compare to Group A. Post-operative radiograph revealed intact implants with evidence of significant bridging callus and better radiographic healing grade in Group B as compare to Group A. Functional outcome score was significantly more in Group B as compare to Group A. The post-operative complication comprised of loosening and distal migration of pin in one case of Group A. The haematobiochemical parameters were within the physiological range and change in the mean values between the groups were statistically non-significant. For the physiological and anatomical repair of the fracture site, the titanium elastic nailing was found to be superior method than End threaded positive profile intramedullary pinning.Titanium elastic nailing resulted in early weight bearing and limb use along with better ability to withstand the stresses of weight bearing.ThesisItem Open Access Studies on regenerative potential of bone marrow derived differentiated osteoblasts and their gene expression in fracture repair in rabbit model(G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand), 2021-03) Bhatt, Jyotsana; Jadon, Narendra S.The present study was conducted to evaluate regenerative potential of bone marrow derived differentiated osteoblast along with β-TCP scaffold in augmenting fracture healing in rabbits and to delineate expression profile of various genes involved in fracture repair as molecular markers of healing via Real time-PCR. During first phase of study isolation and propagation of bone marrow derived MSCs, its differentiation into osteoblasts and their characterization utilizing specific techniques was done followed by second phase of clinical evaluation of therapeutic potential of undifferentiated MSCs and differentiated osteoblasts in fracture repair in rabbit model. Thirty six rabbits used in this study were randomly divided into four equal groups (A, B, C and D) having 9 animals each. Animals were anaesthetized with xylazine @ 6mg/kg followed by ketamine @ 60mg/kg, intramuscularly and a 7mm segmental defect was created in mid diaphysis of left radius bone. Animals of different groups were subjected to different treatment protocols. Animals of group I kept as negative control and only external splint was applied. In animals of group II, only β- TCP implant was used to bridge the segmental defect. Animals of group III were treated with undifferentiated MSCs along with β- TCP implant and animals of group IV were treated with differentiated osteoblasts along with β- TCP implant. Quality of fracture repair was assessed by clinical parameters, radiography, estimation of biochemical parameters, gross evaluation of test bone, histopathology, histochemistry using Van Gieson’s staining, flourochrome labelling, scanning electron microscopy and molecular assessment of bone turnover markers via Real time-PCR. Radiographic findings, gross evaluation of test bone and histopathology revealed that the key criteria for evaluating bone healing like level of callus formation, bony union between cut ends and bone remodelling were significantly (P<0.05) prominent in animals of group IV treated with differentiated osteoblasts along with β-TCP implant closely followed by animals of group III treated with undifferentiated MSCs along with β-TCP implant. Biochemical parameters showed significant (P<0.05) increase in its level on day 30 postoperatively in all the groups, however these values returned to normal level at day 45 in group III and IV signifying faster pace of healing in stem cell treated groups. Tetracycline labelling and scanning electron microscopic study of regenerated tissue from different groups of animals revealed significantly (P<0.05) superior quality of healing in animals of group IV. Gene expression profiling showed upregulation of osteopontin and osteocalcin genes, associated with remodelling, in animals of group IV whereas genes like alkaline phosphatise and collagen type-1, associated with proliferative stage of fracture healing, were upregulated in animals of group I. Results of this study reveals that animals treated with osteogenically predifferentiated MSCs (osteoblasts) showed rapid and superior quality healing of critical sized segmental bone defects as compared to animals treated with undifferentiated MSCs, β-TCP implant alone and control animals.
