Acellular and glutaraldehyde-preserved tendon allografts for reconstruction of superficial digital flexor tendon in bovines: Part I--Clinical, radiological and angiographical observations

Sixteen tenorrhaphies were performed at the mid-metatarsal region in eight buffalo calves under lignocaine epidural analgesia. A 2 cm long gap was created in the superficial digital flexor (SDF) tendon and immediately repaired with acellular grafts in animals of group I, 1% glutaraldehyde-preserved tendon allografts in group II. In group III, the defect was repaired with autografts. This group served as control. The contralateral limb in each animal was operated after an interval of 60 days and the animals underwent the same procedure according to the designed groups. Diclofenac sodium and Enrofloxacin was given post-operatively for 5 days. Clinical examination revealed significant increase (P < 0.05) in rectal temperature, heart and respiratory rate for 3-4 postoperative days in all the animals. Mild pain and exudation as well as early restoration of tendon gliding movements and weight-bearing were observed earlier in group I in comparison with group II. Air-tendograms revealed early organization, minimal adhesion formation and lesser thickening of tendon at the reconstructive site in the acellular group whereas in the glutaraldehyde group dense homogenous swelling with adhesions was seen along the flexors. Angiography on day 30 showed that the area of proximal and distal host tendon graft junction appeared hypervascularized, whereas the area occupied by the graft appeared relatively less vascularized. Normal vascularization was observed on day 90 in all the three groups.     

Carbon fibres and plasma-preserved tendon allografts for gap repair of flexor tendon in bovines: gross,microscopic and scanning electron microscopic observations

The efficacy of carbon fibres and plasma-preserved tendon allografts for gap repair in the superficial digital flexor tendon in the mid-metatarsal region was evaluated in 12 crossbred calves. Experimental tenectomies were performed, followed by implantation of carbon fibres in group I (12 legs) and plasma-preserved tendon allografts in group II (12 legs). Gross observations in group I showed filling of the defect with granulation tissue with more vascularity on day 7, which was less prominent at day 14. On day 30, the neotendon formed was slightly thicker and comparable to normal tendon in appearance and texture. On day 90, it exhibited all the characteristics of a fully developed tendon. Whereas, in group II increased vascularity at the site and encapsulation of the graft with connective tissue in early periods was observed. The gap between graft and host was filled with fibrous connective tissue. Peritendinous adhesions were maximum on day 7 which were gradually reduced in both groups. Microscopically, an acute inflammatory reaction in the periphery of carbon fibres was observed on day 7. Immature fibroblasts were arranged in a haphazard pattern at this stage. By day 14, numerous newly formed capillaries and comparatively more mature fibroblasts were present in between and around the carbon fibres which were aligning parallel to the longitudinal axis of the tendon. By day 30 the healing tissue exhibited longitudinal orientation of collagen fibres and was at a more advance stage of maturation. By day 90, the neotendon formed simulated the picture of normal tendon. In the grafted tendon group, there was normal healing tissue at the functional sites between host and grafted tendon. The fibroblastic activity appeared to be both extrinsic and intrinsic in origin. The connective tissue had invaded the graft to a variable distance and there was resorption of graft which was replaced by newly formed connective tissue on day 90. Scanning electron microscopic observation revealed formation of neotendon between carbon fibre strands, resulting in thickening of the implant. In later stages parallel collagen fibres resembling normal tendon were observed in both groups.     

Acellular dermal graft for repair of abdominal wall defects in rabbits

Sixteen clinically healthy New Zealand white rabbits of either sex were divided into 2 equal groups (I and II) of 8 animals each. Under thiopental sodium (2.5%) anaesthesia a 2 x 3 cm full-thickness abdominal wall defect in the mid-ventral abdominal wall was created and repaired with an acellular dermal graft (ADG) in all the animals of group I (test group). In animals of group II (control group) a full-thickness linear midline abdominal muscular wall incision was made and repaired with a continuous suture pattern using 2-0 nylon.     

Carbon fibres and plasma-preserved tendon allografts for gap repair of flexor tendon in bovines: clinical,radiological and angiographical observations

Twenty-four tenorrhaphies were performed at the mid-metatarsal region in 12 crossbred calves under xylazine-ketamine spinal analgesia. A 2.5-cm long gap was created in the superficial digital flexor (SDF) tendon and immediately repaired with carbon fibres in the animals of group I and with plasma-preserved tendon allografts in group II. Clinical examination revealed a slight increase (P > 0.05) in rectal temperature, heart and respiratory rate for 2-4 days postoperation in both groups. Milder pain and exudation as well as earlier restoration of tendon gliding movements and weight bearing were observed in group I as compared to group II. Air-tendogram in the carbon fibre group on day 30 revealed restoration of continuity across the defect of the tendon. whereas, in the allograft group, a dense homogeneous swelling was seen along the flexors. Regression of peritendinous adhesions and swelling at the reconstructed site at later stages was seen in both groups. Angiography showed hypervascularization at the reconstructed site on day 14 in the carbon fibre group, however, in the allograft group the site appeared to be relatively avascular. On days 30 and 90, blood vessels were normally organized in both groups.     

Gross and histomorphological observations following homologous deep frozen tendon grafting in equines

Experimental tenectomies were performed in the mid metacarpal region of the superficial digital flexor (SDF) tendon while the deep digital flexor (DDF) tendon was traumatized in 10 donkeys. In group I (n=8) the deep frozen tendons were used for grafting where as in group II (n=2) no grafting was done. Amniotic membrane casing was performed around grafted SDF and traumatized DDF tendons in group I only. Gross and histomorphological observations were recorded on the last day of the experiment, i.e., on the 15th (n=1), 30th (n=2), 60th (n=2), 90th (n=2) and 120th (n=1), post-operative days (POD) in group I and on the 60th (n=1) and 120th (n=1) POD in group II. Gross intraoperative observations suggested the presence of only thin filamentous adhesions in all but three animals at 30 (two animals) and 60 (one animal) days which were present only in the limited area of grafted tendon as compared to tenectomized SDF tendon that showed thick fibrous adhesion and thickening at both the cut ends. However, DDF tendon showed no adhesions in all except one animal of group I as compared to group II where adhesions were noticed in both the animals. Histological examination suggested an increase in fibroblastic activity at 2 weeks after grafting. Extrinsic and intrinsic healing response was observed in grafted tendons. Inflammatory reaction was reabsorbed by one month. The bundle formation was observed from two to three months. The histological structure appeared near normal by 4 months. Observation suggested that homologous tendon grafts remain viable by all histological criteria and healing was at a comparatively more advanced stage than tenectomized tendon. However, the traumatized DDF tendon healed completely by two months. It is concluded that the tendon healing was more rapid in animals where deep frozen grafts had been used.     

A preliminary study on the effect of ultrasound therapy on the healing of surgically severed achilles tendons in five dogs

This study was conducted on the left Achilles tendon in five clinically normal dogs. The Achilles tendon was surgically exposed and severed 3-4 cm proximal to the point of its insertion. Tenorrhaphy was undertaken by the application of three sutures on the various tendon units of the Achilles tendon using single locking-loop sutures with polyamide no. 1-0. The superficial digital flexor tendon was sutured with catgut using two horizontal mattress sutures. No ultrasound therapy was used in the animals of group I (control). Ultrasound therapy was given to the animals of group II (treated) starting from the third day post-operatively at 0.5 W/cm2 for 10 min daily for 10 days. A cortical screw was used for immobilization of the tibiotarsal joint which was removed 4 weeks after tenorrhaphy. Post-operatively, healing of the Achilles tendon was monitored using clinical observations, ultrasonography, gross and histomorphological observations at various intervals up to 120 days in both groups. Clinically, the dogs showed significant lameness for the first 4-5 days, which disappeared earlier in the ultrasound-treated (group II) animals than the controls (group I). Extension and flexion of the hock joint were found to be near normal at 6 weeks after the repair of the Achilles tendon. Ultrasonography showed anechoic to hypo-echoic echo-texture on days 3 and 7 after repair. By day 40, the echo-texture started to improve to hypo-echoic in group II, but in group I anechoic areas were still observed. However, the tendon showed near normal mottled hypo- to hyper-echoic texture in both groups by day 120. Gross observations suggested that the Achilles tendon in group II showed comparatively fewer adhesions than in group I animals. Histologically, in group II (treated), on day 40, the union was comparatively better without any inflammatory reaction. Bundle formation had begun in the ultrasound-treated animals which was not observed in the control animals. By day 90, more compact parallel bundle formation had taken place with minimum cellularity. Bundle formation was in its advanced stage in the treated animals. By day 120, the tendon tissue was comparatively acellular and looking like a normal tendon. The use of the cortical screw provided good immobilization and ultrasound therapy at 0.5 W/cm2 enhanced the Achilles tendon healing in dogs.     

Treatment of chronic complete rupture of achilles tendon in three dogs with locking loop suture and own suture technique

The present case report describes three heavy dogs, Caucasian Shepherd breed, with chronic complete traumatic rupture of Achilles tendon, which was repaired with two types of sutures. Surgical treatment involved shortening the Achilles tendon, and using a locking loop suture technique and own suture technique with four buttons. Additionally, positional screw and cast supported the sutures. Twelve months postoperatively no discomfort and lameness were observed on clinical examination. This paper shows that treatment of chronic complete rupture of Achilles tendon with locking loop suture and own suture technique gives good results.     

Immunohistochemical and scanning electron microscopic comparison of the collagen network constructions between pig, goat and chicken livers

The distribution and three-dimensional architecture of collagen fibers were compared between pig, goat and chicken livers. Immunohistochemical staining revealed that collagen type I was identified in the interlobular connective tissue region and intralobular areas in pigs and goats. Type III collagen was also identified in the interlobular connective tissue region and intralobular sinusoidal walls. In the chicken liver, only the circumference region of the vessels was immunostained with collagen type I and III antibodies and the interlobular connective tissue wall could not be distinguished clearly. In the intralobular region, collagen type I antibody immunoreacted around the hepatic cells but collagen type III antibody immunoreacted weakly. In the NaOH macerated specimen, well-developed collagen bundles formed the prominent interlobular walls in pigs. In contrast, the wall in the goat liver comprised a thin layer of the bundles. In the chicken liver, there were no notable collagen septa between lobules. The intralobular collagen construction was quite different between the animals, indicating a fragile collagen fibril networks in pigs, a robust framework in goats and dense fabric-like septa in chickens. These results indicate that the distinct collagen frameworks may contribute to the histological strength of the livers in each of the animal species.     

Effect of preservation methods on the performance of bovine pericardium graft in a rat model

This study investigates the effect of preservation methods on the performance of bovine parietal pericardium grafts in a rat model. Mid-ventral full thickness abdominal wall defects of 3 x 2.5 cm in size were created in 90 male Sprague-Dawley rats (300-400 g), which were divided into three groups of 30 rats each. The abdominal defects of group one and two were repaired with lyophilized and glycerolized bovine pericardium grafts, while the defects of group three were repaired with expanded polytetrafluoroethylene (ePTFE) Mycro Mesh as a positive control. Another group of 30 rats underwent sham operation and was used for comparison as negative control. Each group of rats (n = 30) was divided into five subgroups (n = 6) and killed at 1, 3, 6, 9 and 18 weeks post-surgery for gross and morphological evaluations. The rats tolerated the surgical procedure well with a total mortality of 0.05%. No serious post-operative clinical complications or signs of rejection were encountered. Adhesions between the grafts and the underlying visceral organs observed in the study were mostly results of post-surgical complications. Glycerol preservation delayed degradation and replacement of the grafts, whereas lyophilization caused early resorption and replacement of the grafts. The glycerolized grafts were replaced with thick dense fibrous tissue, and the lyophilized grafts were replaced with thin loose fibrous tissue. The healing characteristic of the bovine pericardium grafts was similar to those of the sham-operated group, and quite different from those of the ePTFE Mycro Mesh. The outcome of the present study confirmed the superiority of glycerolized bovine pericardium grafts over its lyophilized counter part.     

A light and scanning electron microscopic study of the epididymis active state of the endemic Mexican rodent Peromyscus winkelmanni (Carleton) (Rodentia: Muridae)

The macroscopic and microscopic anatomy of the epididymis of the sexually mature Peromyscus winkelmanni (carleton) was examined using light and scanning electron microscopy. The epididymis was divided into three regions: caput, corpus and cauda. The epididymal duct was lined with columnar and cubic epithelium with stereocilia and covered by a muscular connective tissue sheath. Capillaries appear to penetrate directly into the epithelium from the underlying connective tissue in the initial segment. The epididymal epithelium presents four cell types: principal, basal, apical and clear cells. Based on morphological differences (height of epithelial cells, length of the stereocilia, luminal area, larger diameter and spermatic index), the epididymis of P. winkelmanni, presents seven zones. The stereocilia of the epididymal ducts of zones I, II, IV and V are thick and tall, while in zone III they are thin and short. The stereocilia in zone VI are thin, while in zone VII they are short but thick. The secretory products observed in the lumen of the epididymal ducts have vesicular, granular and fibrous form in the seven zones. This study contributes to an understanding of the morphofunctional features of the epididymis in sperm maturation in a species that shows seasonal reproductive activity.     

Cervical vertebral interbody fusion in the horse: a comparative study of bovine xenografts and autografts supported by stainless steel baskets

A modified form of the Cloward technique for anterior cervical fusion in human beings was used in the application of different grafts for evaluation of their effectiveness in stabilizing equine cervical vertebrae. Results of bovine xenograft implants in 8 horses were compared with results of stainless steel baskets (SSB) packed with cancellous autogenous bone in 8 horses. Graft material was incorporated in all cases. Evidence of graft rejection was not present. Both forms of implants decreased the mobility of the intervertebral space in which they were implanted. A fibrous connective tissue union was prevalent after xenograft implantation. Osseous union was prevalent after implantation of the SSB. Reduction in range of motion was greatest at those sites with the SSB.     

An evaluation of two autologous tendon grafting techniques in ponies

OBJECTIVE: To compare the healing and mechanical strength of a multiple split autologous tendon graft (MG) to a whole autologous tendon graft (WG) in the deep digital flexor tendon of ponies. STUDY DESIGN: In vitro evaluation of two different tendon-grafting techniques. ANIMALS OR SAMPLE POPULATION: Six ponies of mixed gender and age. METHODS: Tenotomies performed in forelimb deep digital flexor tendons (DDFT) distal to the insertion of the accessory ligament (AL-DDFT) were repaired with free autologous grafts from the hindlimb lateral digital extensor tendon (LDET). Grafts were either whole (WG) or split into three longitudinal strips (multiple graft, MG). Tendons and graft sites were collected and loaded (2.54 cm/s) to failure at either 4 or 8 weeks after surgery. Cross-sectional area was determined by both impression cast (IC) and an inkblot (IB) method. Tissue maturity and inflammation were evaluated by microscopy. RESULTS: Gap formation was a consistent finding in all repair sites. No statistical differences were found in healing or mechanical variables between MG and WG techniques. The failure stress for the 8-week repairs (15.51 +/- 3.1 MPa IB and 11.73 +/- .77 MPa IC, 16.13 +/- 2.2 MPa IB and 10.22 +/- .76 MPa IC for MG and WG, respectively) were significantly greater (P <.0005) than for 4-week repairs (3.71 +/- 1.7 MPa IB and 2.68 +/- 1.44 MPa IC, 2.81 +/- 1.46 MPa IB and 2.3 +/- 1.7 MPa IC for MG and WG, respectively). The repair tissue was more mature (P <.05) at 8 weeks than at 4 weeks, but there was no significant difference in inflammatory responses at 4 and 8 weeks. CONCLUSIONS: There was a sixfold increase in strength between 4 and 8 weeks of healing, but no significant difference in healing or strength between the MG and WG techniques. CLINICAL RELEVANCE: In ponies, autologous tendon grafting contributes to a strong repair during the early convalescent period, but splitting a tendon graft seemingly offers no appreciable advantage over use of a whole graft.     

Histology and immunohistochemistry study of ovine tendon grafted with cBMSCs and BMMNCs after collagenase-induced tendinitis

OBJECTIVES: The aim of this study was to compare the regeneration abilities of cultured bone marrow mesenchymal cells (cBMSC) and bone marrow mononuclear cells (BMMNC) with fibrin glue, saline solution and sham control in collagenase-induced tendinitis of the Achilles tendon in sheep. METHODS: Six sheep were recruited randomly to each group: cBMSC, BMMNC, fibrin, saline and sham control. Each group received the relative treatment two weeks after inducing lesions (T(0)). After eight weeks (T(8)) of treatment, the tendons were harvested and evaluated for histomorphology, Collagen type I, III, Cartilage Oligomeric Matrix Protein (COMP) and CD34 positive cells expression. RESULTS: Histology and immunohistochemistry showed similar capabilities of cBMSC and BMMNC to restore the architecture of fibres and Extra Cellular Matrix (ECM), with a high expression of collagen type I and COMP and a very low expression of collagen type III in treated tendons. The complete architectural disruption of fibres, dramatic reduction of collagen Type I and COMP expression and increase collagen type III expression were commonly observed in tendons treated with fibrin or saline only. The presence of CD34 positive cells was appreciable in the BMMNC group while few cBMSC showed this cluster of differentiation, not expressed in tendons treated with fibrin or saline. CLINICAL SIGNIFICANCE: The data in this study show the efficacy of cBMSC and BMMNC in regenerating tendon tissue after collagenase-induced tendinitis.     

End-to-End Colonic Anastomosis A Comparison of Techniques in Normal Dogs

Colonic resections were repaired in normal dogs by crushing suture, inverting suture, or staple anastomosis techniques. Dogs were sacrificed 2, 4, 7, 10, 14, 21, and 28 days following surgery. Bacterial cultures, breaking strength measurement, adhesion formation, and gross and histopathologic examinations were made. Anastomoses closed by staples showed the least tissue reaction, least neutrophilic infiltration, most mature fibrous connective tissue, most normal arrangement of muscle layers, and fewest numbers of mucoceles and necrotic areas. The staple and crushing anastomosis techniques caused significantly less reduction in luminal radius than did the inverting anastomosis technique. Bacteria were isolated least frequently and adhesions were least severe following staple anastomosis, though these results were not statistically significant. Stapled anastomosis appeared to result in a higher tensile strength of the wounds at the anastomotic sites compared with the other techniques studied.     

Experimental rickets in broilers: gross, microscopic, and radiographic lesions. III. Vitamin D deficiency

Day-old broiler chicks were fed a vitamin D-deficient diet for 30 days. Gross, microscopic, and radiographic examinations of the proximal tibiotarsus were done at 2, 3, and 4 weeks of age. In 2-week-old chicks, there was variable lengthening and disorganization of the proliferating-prehypertrophied zone (P-PHZ), resorption of cartilage spicules in the degenerating hypertrophied zone and primary spongiosa, and lengthening of bone spicules in the secondary spongiosa; many bone surfaces were lined with increased osteoid. In 3-week-old chicks, there was a tendency for the P-PHZ to decrease in relative length and for cartilage spicules to increase in length. In 4-week-old chicks, however, there was marked lengthening of the P-PHZ, resorption of cartilage spicules, and replacement of the metaphysis with irregularly oriented islands of woven bone, osteoid, and loose fibrous connective tissue.     

Healing of surgically created defects in the equine superficial digital flexor tendon: collagen-type transformation and tissue morphologic reorganization

Full-thickness defects were surgically created in the superficial digital flexor tendons of the front limbs of 20 horses. Tissues formed within the defect were evaluated histologically, and the collagen composition of the tissue was determined by immunofluorescence. Transformation occurred from loose fibrillar areas of types I and III collagen and pericellular types IV and V collagen to dense bundles of type I collagen fibers. Loose fibrillar areas of types I and III collagen were present after 24 weeks. Histologically, in horses killed after 2 weeks, the tissue within the defect was a randomly oriented mass of fibrovascular tissue. In horses killed after 24 weeks the tissue within the defect resembled normal tendon tissue.     

A Comparison of Repair Methods for Gap Healing in Equine Flexor Tendon

In nine horses (18 forelimbs), a 3 cm section of superficial digital flexor tendon was removed and the tendons were repaired with immobilization for 6 weeks and (1) no suture (n = 6); (2) a double locking loop tenorrhaphy with carbon fiber (n = 6); or (3) a double locking loop tenorrhaphy with size 2 nylon suture (n = 6). Clinical assessment, gross evaluation, morphometry, histology, and mechanical testing were performed on two limbs from each treatment group at weeks 6, 12, and 24. At weeks 6 and 12, the unsutured tissue was less mature than the tissue sutured with nylon. By week 24, the carbon fiber repair had breaking stress (mean, 12.5 M Pa) similar to the unsutured repair (mean, 10.6 M Pa). There was necrosis and a granulomatous foreign body reaction around the carbon fiber. The nylon suture repair had significantly greater strength per unit area (mean breaking stress, 20.4 M Pa) because of a smaller area of repair tissue in the gap (mean, 3.6 cm2). At week 24, the nylon suture repair tissue was the most mature with the least inflammation of the three repair methods. A nylon double locking loop suture was the preferred method for equine flexor tenorrhaphy of a tendon gap because of greater breaking stress, histologic maturity, biocompatibility, and the adequate functional and cosmetic outcome.     

Comparison of different bronchial closure techniques following pneumonectomy in dogs

The comparison of the histologic healing and bronchopleural fistula (BPF) complications encountered with three different BS closure techniques (manual suture, stapler and manual suture plus tissue flab) after pneumonectomy in dogs was investigated for a one-month period. The dogs were separated into two groups: group I (GI) (n = 9) and group II (GII) (n = 9). Right and left pneumonectomies were performed on the animals in GI and GII, respectively. Each group was further divided into three subgroups according to BS closure technique: subgroup I (SGI) (n = 3), manual suture; subgroup II (SGII) (n = 3), stapler; and subgroup III (SGIII) (n = 3), manual suture plus tissue flab. The dogs were sacrificed after one month of observation, and the bronchial stumps were removed for histological examination. The complications observed during a one-month period following pneumonectomy in nine dogs (n = 9) were: BPF (n = 5), peri-operative cardiac arrest (n = 1), post-operative respiratory arrest (n = 1), post-operative cardiac failure (n = 1) and cardio-pulmonary failure (n = 1). Histological healing was classified as complete or incomplete healing. Histological healing and BPF complications in the subgroups were analyzed statistically. There was no significant difference in histological healing between SGI and SGIII (p = 1.00; p > 0.05), nor between SGII and SGIII (p = 1.00; p > 0.05). Similarly, no significant difference was observed between the subgroups in terms of BPF (p = 0.945; p > 0.05). The results of the statistical analysis indicated that manual suture, stapler or manual suture plus tissue flab could be alternative methods for BS closure following pneumonectomy in dogs.     

Histology and ultrastructure of the developing superficial digital flexor tendon in rabbits

This study was designed to investigate the developmental changes in the superficial digital flexor tendon (SDFT) of the white New Zealand rabbit, from 5 to 7 days pre-natal to 112 days post-natal (PN), at histological and ultrastructural levels. The tendons changed from being highly cellular with 13079 ± 2538 cell/mm² and little directional organization to a longitudinally oriented, predominantly connective tissue structure with relatively few, mature tenocytes (2384 ± 365 cell/mm²). Fibrillogenesis was seen in isolated vacuoles of fibroblast cytoplasm during fetal life as well as after birth, but less frequently with increasing age. At the ultrastructural level, there was a progressive increase in the mean diameter of collagen fibrils with age throughout the population , until PN day 28. A bimodal distribution of collagen fibril size was first observed on PN day 56 while at day 112, the fibrils were fully differentiated and showed a multimodal size distribution. The development of elastic fibres preceded that of collagen fibres and accompanied progressively more marked sinusoidal crimping in collagen fibres on PN days 7 and 14. The tendons of older animals became less tightly crimped. The cells of the epi- and endotenon were less mature and relatively undifferentiated compared with the cells in the body of the tendon of the same age, which might explain their ability to initiate healing of tendon injuries in older animals. In conclusion, extensive changes were observed in tendons during growth and maturation, with diameters of collagen fibrils, tissue orientation and cellularity being the parameters affected, probably due largely to increased physical activity.