Luca Lacitignola

Histological and Immunohistochemical Evaluation of Autologous Cultured Bone Marrow Mesenchymal Stem Cells and Bone Marrow Mononucleated Cells in Collagenase-Induced Tendinitis of Equine Superficial Digital Flexor Tendon

The aim of this study was to compare treatment with cultured bone marrow stromal cells (cBMSCs), bone marrow Mononucleated Cells (BMMNCs), and placebo to repair collagenase-induced tendinitis in horses. In six adult Standardbred horses, 4000 IU of collagenase were injected in the superficial digital flexor tendon (SDFT). Three weeks after collagenase treatment, an average of either 5.5 × 106 cBMSCs or 1.2 × 108 BMMNCs, fibrin glue, and saline solution was injected intralesionally in random order. In cBMSC- and BMMNCS-treated tendons, a high expression of cartilage oligomeric matrix protein (COMP) and type I collagen, but low levels of type III collagen were revealed by immunohistochemistry, with a normal longitudinally oriented fiber pattern. Placebo-treated tendons expressed very low quantities of COMP and type I collagen but large numbers of randomly oriented type III collagen fibers. Both cBMSC and BMMNCS grafts resulted in a qualitatively similar heling improvement of tendon extracellular matrix, in terms of the type I/III collagen ratio, fiber orientation, and COMP expression.

Cell Therapy for Tendon Repair in Horses: An Experimental Study

Crovace, A., Lacitignola, L., De siena, R., Rossi, G. and Francioso, E., 2007. Cell therapy for tendon repair in horses: An experimental study. Veterinary Research Communications, 31(Suppl. 1), 281–283

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.

Low respiratory rate plus minimally invasive extracorporeal Co2 removal decreases systemic and pulmonary inflammatory mediators in experimental Acute Respiratory Distress Syndrome.

Objective:The Acute Respiratory Distress Syndrome Network protocol recommends limiting tidal volume and plateau pressure; it also recommends increasing respiratory rate to prevent hypercapnia. We tested a strategy that combines the low tidal volume with lower respiratory rates and minimally invasive CO2 removal. Subjects:Ten lung-damaged pigs (instilled hydrochloride). Interventions:Two conditions randomly applied in a crossover fashion: the Acute Respiratory Distress Syndrome Network protocol and the Acute Respiratory Distress Syndrome Network protocol plus lower respiratory rate plus minimally invasive Co2 removal. A similar arterial Co2 partial pressure was targeted in the two conditions. Measurements and Main Results:Physiological parameters, computed tomography scans, plasma and bronchoalveolar lavage concentrations of interleukin-1&bgr;, interleukin-6, interleukin-8, interleukin-10, interleukin-18, and tumor necrosis factor-&agr;. During the lower respiratory rate condition, respiratory rate was reduced from 30.5 ± 3.8 to 14.2 ± 3.5 (p < 0.01) breaths/min and minute ventilation from 10.4 ± 1.6 to 4.9 ± 1.7 L/min (p < 0.01). The extracorporeal device removed 38.9% ± 6.1% (79.9 ± 18.4 mL/min) of CO2 production. During the lower respiratory rate condition, interleukin-6, interleukin-8, and tumor necrosis factor-&agr; concentrations were significantly lower in plasma; interleukin-6 and tumor necrosis factor-&agr; concentrations were lower in bronchoalveolar lavage, whereas the concentrations of the other cytokines remained unchanged. Conclusion:The strategy of lower respiratory rate plus minimally invasive extracorporeal CO2 removal was feasible and safe and, as compared with the Acute Respiratory Distress Syndrome Network protocol, reduced the concentrations of some, but not all, of the tested cytokines without affecting respiratory mechanics, gas exchange, and hemodynamics.

Survival of bone marrow mesenchymal stem cells labelled with red fluorescent protein in an ovine model of collagenase-induced tendinitis

OBJECTIVE The aim of this study was to track the survival and efficacy of allogeneic bone marrow mesenchymal stem cells (BM-MSC) marked with red fluorescent protein (BM-MSCRFP) in an ovine model of collagenase-induced tendinopathy. METHODS Bone marrow was harvested from one donor sheep and BM-MSC were isolated, cultivated and transfected with red fluorescent protein (BM-MSCRFP). Collagenase was injected into both Achilles tendons in the remaining nine sheep. After two weeks the left tendon was injected with a solution of 6 x 10⁶ BM-MSCRFP and fibrin glue, while only fibrin glue was administered to the contra-lateral tendon in each sheep. After three, four and six weeks the tendons were harvested and evaluated for morphology, collagen I deposition, presence of CD34+ cells, and fluorescent labelled BM-MSC. RESULTS We demonstrated that delivery of BM-MSC into tendon lesions had positive effects on the injured tendons. The BM-MSCRFP survived at three, four and six weeks after treatment, leading to better quality healing of tendons as compared to the controls, where no labelled cells were detected. Interestingly, we demonstrated high expression of CD34+ cells in tendons that had been treated with BM-MSCRFP. CLINICAL RELEVANCE Mesenchymal stem cell allografts have a positive effect on tendon healing and local injection of BM-MSC directly into the tendon allows the homing of BM-MSC for good efficiency of engraftment.

A comparison of subarachnoid buprenorphine or xylazine as an adjunct to lidocaine for analgesia in goats.

OBJECTIVE To test the hypothesis that subarachnoid administration of buprenorphine and lidocaine provides more intense and longer lasting perioperative analgesia with less side effects than xylazine and lidocaine in goats. STUDY DESIGN Randomized, blinded, controlled study. STUDY ANIMALS: Ten healthy female goats randomly assigned to two groups of five animals each. METHODS After sedation with acepromazine (0.1 mg kg(-1)) intravenously (i.v.), lidocaine 2% (0.1 mL kg(-1)) combined with either xylazine (0.05 mg kg(-1); Group X) or buprenorphine (0.005 mg kg(-1); Group B) were injected intrathecally at the lumbo-sacral junction prior to stifle surgery. Electrocardiogram, heart rate, direct systolic, mean, and diastolic arterial blood pressures, rectal temperature and arterial blood gases were recorded as were post-operative sedation and pain scores using a visual analogue and numeric rating scale, respectively. Data were analyzed with one-way ANOVA for repeated measures, one-way anova, Friedmans and Kruskal-Wallis tests as necessary (p < 0.05). RESULTS Surgery was successfully performed under both analgesia protocols. Total pain and sedation scores were significantly lower in the B as compared with X group from 3-24 hours and 30-120 minutes, respectively after subarachnoid drug administration (SDA). Heart rate and arterial blood pressures decreased post SDA and were consistently lower in X versus B (p < 0.05). In B arterial blood gas parameters did not change post SDA, but in group X PaCO(2) increased slightly within 15 minutes of SDA and remained elevated for at least 3 hours (p < 0.05). CONCLUSION In these goats intrathecal administration of buprenorphine and lidocaine produced more profound and longer lasting analgesia with less sedation and hemodynamic and respiratory impairment than xylazine with lidocaine. CLINICAL RELEVANCE In these goats undergoing hind limb surgery, subarachnoid buprenorphine/lidocaine offered more intense and longer lasting analgesia than a xylazine/lidocaine combination, with less sedation and impairment of cardiopulmonary function.

Plasma Processing of Scaffolds for Tissue Engineering and Regenerative Medicine

Abstract Plasma processes are largely employed in the biomedical field for different kind of materials. In particular, in tissue engineering, biomaterials need to be totally integrated with biological systems in order to be employed as substitutes of artificial prostheses. Since most materials do not allow a correct integration with the biological environment, plasma processes have been demonstrated to be very versatile in altering the material surface properties in order to improve the biocompatibility of materials. The challenge is to plasma modify 3D scaffolds in order to be used for in vivo regeneration of human tissues. The correct 3D biointegration inside living tissues is the crucial objective, towards which many aspects are directed, from the material engineering to its surface modification and affinity with the biological environment. In this paper, the advances in low pressure plasma processes, applied to both 2D rigid substrates and 3D porous structures, are discussed. Further an in vivo experiment in ovine animals using plasma processed 3D scaffolds is illustrated.

Computed Tomographic Tenography of Normal Equine Digital Flexor Tendon Sheath: An Ex Vivo Study.

Aim of this study was to document the normal computed tomographic tenography findings of digital flexor tendon sheath. Six ex vivo normal equine forelimbs were used. An axial approach was used to inject 185 mg/mL of iopamidol in a total volume of 60 mL into the digital flexor tendon sheaths. Single-slice helical scans, with 5 mm thickness, spaced every 3 mm, for a pitch of 0.6, and with bone algorithm reconstruction, were performed before and after injections of contrast medium. To obtain better image quality for multiplanar reconstruction and 3D reformatting, postprocessing retroreconstruction was performed to reduce the images to submillimetre thickness. Computed tomographic tenography of digital flexor tendon sheaths could visualize the following main tendon structures for every forelimb in contrast-enhanced images as low densities surrounded by high densities: superficial digital flexor tendon, deep digital flexor tendon, manica flexoria, mesotendons, and synovial recess. Results of this study suggest that computed tomographic tenography can be used with accuracy and sensitivity to evaluate the common disorders of the equine digital flexor tendon sheath and the intrathecal structures.

WHICH AIRWAY PRESSURE SHOULD BE APPLIED DURING BREATH-HOLD IN DOGS UNDERGOING THORACIC COMPUTED TOMOGRAPHY?

This randomized controlled trial study aimed to identify the optimal positive pressure (PP) level that can clear atelectasis while avoiding pulmonary hyperinflation during the breath-hold technique in dogs undergoing thoracic computed tomography (CT). Sixty dogs affected by mammary tumors undergoing thoracic CT for the screening of pulmonary metastases were randomly assigned to six groups with different levels of PP during the breath-hold technique: 0 (control), 5 (PP5), 8 (PP8), 10 (PP10), 12 (PP12), and 15 (PP15) cmH2 O. The percentage of atelectatic lung region was lower in the PP10 (3.7 ± 1.1%; P = 0.002), PP12 (3.4 ± 1.3%; P = 0.0001), and PP15 (2.8 ± 0.9%; P = 0.006) groups than in the control group (5.0 ± 2.3%), and the percentage of poorly aerated lung region was lower in the PP8 (15.1 ± 2.6%; P = 0.0009), PP10 (13.0 ± 2.0 %; P = 0.002), PP12 (13.0 ± 2.2 %; P = 0.0002), and PP15 (11.1 ± 1.9%; P = 0.0002) groups than in the control group (19.8 ± 5.0). The percentage of normally aerated lung region, however, was higher in the PP10 (79.7 ± 4.1%; P = 0.005), PP12 (79.8 ± 5.1%; P = 0.0002), and PP15 (80.2 ± 4.9%; P = 0.002) groups than in the control group (73.4 ± 6.6%). A PP of 10-12 cmH2 O during the breath-hold technique should be considered to improve lung aeration during a breath-hold technique in dogs undergoing thoracic CT.

Intraoperative contrast echocardiography to verify the surgical occlusion of a single extrahepatic portosystemic shunt in a dog.

This report details a bubble echocardiographic study carried out during the surgical treatment of a congenital single extrahepatic portosystemic shunt (PSS) in a Labrador Retriever. After celiotomy, agitated saline was injected through a jejunal vein and microbubbles appeared rapidly in the right cardiac chambers. The test confirmed the presence of a PSS, helping the surgeon to identify the vessel concerned and to rule out a second shunt. Successively, portography confirmed what the exploratory celiotomy had revealed before with the aid of the bubble study: a single shunt was located between the portal vein and the right renal vein. It was completely ligated, as all the criteria for this solution were met. Intraoperative contrast echocardiography (ICE) was easy to perform, helpful and undemanding. It is proposed here as an intraoperative ancillary test to diagnose all PSS and to confirm successful treatment when complete shunt closure is possible.