Intravenous injection of allogenic canine mesenchymal stem cells in 40 client-owned dogs: a safety assessment in veterinary clinical trials.

BACKGROUND: The aim of this study was to evaluate the adverse effects of allogeneic mesenchymal stem cells (MSCs) transplanted via intravenous infusion in dogs and examine their safety. We performed a retrospective analysis of various clinical assessments, including physical examination, blood tests, and radiographs, and monitored the formation of neoplasms during a 6-month follow-up period in 40 client-owned dogs that received intravenous infusion of adipose tissue-derived MSCs (AT-MSCs) for the treatment of various underlying diseases between 2012 and 2018. RESULTS: No significant adverse effects of MSC therapy were detected by clinical assessment, blood tests, or radiographic examination in the 6-month follow-up period after the first MSC treatment. Additionally no new neoplasms were observed during this period. CONCLUSIONS: To our knowledge, this study is the first to evaluate the safety aspects (≥ 6 months) associated with intravenous allogeneic AT-MSC infusion. These results suggest that allogenic AT-MSC infusion could be a useful and relatively safe therapeutic approach in canines.

Testicular subcutaneous allografting followed by immunosuppressive treatment promotes maintenance of spermatogonial cells in rainbow trout (Oncorhynchus mykiss).

Germ cell transplantation and testis graft represent promising biotechnologies that can be applied for the reproduction of commercial or endangered species. However, mechanisms of rejection from the host immune system might remove the transplanted donor cells/tissues and limit the surrogate production of gametes. In this work, we administered emulsion containing-immunosuppressants to verify whether they are capable to prevent immune rejection and promote survival of testis allografts in rainbow trout. In the first part of this study, we demonstrated in vitro that tacrolimus and cyclosporine were able to affect viability, inhibit leucocyte proliferation, and suppress il2 expression in vitro. In in vivo experiments, both doses of tacrolimus (0.5 and 1.5 mg/kg) and the lower dose of cyclosporine (20 mg/kg) significantly inhibited the expression of il2 in head kidney, three days post-injection. A higher dose of cyclosporine (40 mg/kg) was able to inhibit il2 expression for up to seven days post-injection. In the second part, testis allografts were conducted in fish treated weekly with emulsion containing-tacrolimus. Immunohistochemical, conventional histology, and qRT-PCR (vasa) analysis demonstrated the presence of spermatogonial cells by the fifth week, in animals treated with 0.5 mg/kg of tacrolimus similar as found in autografted group. In the group treated with the highest tacrolimus dose (1.5 mg/kg) and in the non-treated group (without immunosuppressant), no germ cells or their respective markers were detected. il2 expression in head kidney was also suppressed in grafted animals treated with tacrolimus compared to non-treated group. These results suggest that tacrolimus may be a promising immunosuppressant for testis allografts or germ cell transplantation in rainbow trout. Co-administration combining tacrolimus (at lower dose) with other immunosuppressive drugs for inhibiting other activation pathways of the immune system, as performed in human organ transplantation, could be an alternative approach to optimize the immunosuppressive effects in host organisms.

Comparison of diameter and length of subclavian arteries to external jugular veins in variably sized dogs: A cadaveric study.

OBJECTIVE: To evaluate the length and diameter of a left external jugular vein graft as a substitute for the left subclavian artery in the modified Blalock-Thomas-Taussig shunt (mBTTS) in differently sized dogs. STUDY DESIGN: Cadaveric study. ANIMALS: Dog cadavers of three weight categories (10/group): <9.5 kg, 9.5 to 27 kg, and > 27 kg. METHODS: The length and infused external diameters of harvested vessels were measured with vernier calipers and recorded. A matched-pairs t test was used to test the difference in vessel lengths. The agreement in vessel diameters was assessed by using Lin's concordance correlation coefficient (CCC). Pearson's correlation coefficients (CC) were determined for vessel diameter to weight category and vessel length to weight category. RESULTS: The external jugular vein measured longer than the subclavian artery in all dogs (52.0 ± 20.8 mm and 23.0 ± 8.9 mm, respectively), with a mean difference of 28 ± 14.3 mm (P < .001). The mean external infused subclavian and external jugular diameters measured 7.8 ± 2.2 mm and 8.0 ± 2.5 mm, respectively (P = .32). Lin's CCC was 0.87. Pearson's CC were 0.74 in both vessel diameters (P < .001); they were 0.36 and 0.43, respectively, for subclavian artery and external juglar vein length (P < .001). CONCLUSION: Autologous external jugular vein grafts had an external diameter similar to subclavian artery and a significantly longer length in variably sized dogs. CLINICAL SIGNIFICANCE: External jugular vein grafts may be an acceptable graft choice for mBTTS.

Clinical safety of intratesticular transplantation of allogeneic bone marrow multipotent stromal cells in stallions.

Although stem cell therapy is a promising alternative for treatment of degenerative diseases, there are just few reports on the use of stem cells therapy in horse's reproductive system. This study aims to evaluate the effect of intratesticular injection of bone marrow mesenchymal stromal/stem cells (MSCs) in healthy stallions, and its outcome on seminal parameters and fertility. In Experiment 1, 24 stallions were divided into treatment group (TG) and control group (CG). In the TG, an intratesticular application of MSC was performed, and in the CG, only PBS was used. Measurements of testicular volume, surface temperature and Doppler ultrasonography were performed 24 and 48 hr after treatments. Fifteen days after application, the testicles were removed and submitted to histological analysis. In Experiment 2, 3 fertile stallions received similarly treatment with MSCs. Physical examination and sperm analysis were performed weekly during 60 days after treatment, and at the end, semen from one of them was used for artificial inseminations of 6 healthy mares. In Experiment 1, clinical examinations showed no signals of acute inflammation on both groups according to the analysed variables (p > .05). Also, no signal of chronic inflammation was observed on histological evaluation. In Experiment 2, stallions presented no physical alterations or changes in sperm parameters, and a satisfactory fertility rate (83%; 5/6) was observed after AI. The results support the hypothesis that intratesticular application of bone marrow MSCs is a safe procedure, and this could be a promising alternative to treat testicular degenerative conditions.

Production of donor-derived offspring by allogeneic transplantation of spermatogonia in Chinese rosy bitterling†.

Many bitterling species are facing extinction because of habitat destruction. Since cryopreservation of fish eggs is still not available to date due to their large size and high yolk content, long-term and stable storage of bitterling genetic resources is currently not possible. We recently discovered that cryopreservation of early-stage germ cells is possible in several fish species and that functional gametes derived from the frozen materials can be produced through their transplantation to embryonic recipients. However, bitterlings have uniquely shaped eggs and their embryos are extremely fragile, making it difficult to perform germ cell transplantation. Therefore, as a first step, we conducted intra-species spermatogonial transplantation using recessive albino Chinese rosy bitterling as donors and wild-type Chinese rosy bitterling as recipients to develop a system to convert freezable early-stage germ cells into functional gametes, particularly eggs. Approximately 3000 testicular cells were transplanted into the peritoneal cavity of 4-day-old germ cell-less recipient embryos produced by dead end (dnd)-knockdown. At 6 months, ten male recipients and nine female recipients produced gametes. Mating studies with the opposite sex of recessive albino control fish revealed that six males and three females produced only albino offspring, suggesting that these recipients' endogenous germ cells were completely removed by dnd-knockdown and they produced only donor-derived gametes. Thus, we successfully established a germ cell transplantation system in an iconic endangered teleost, bitterling. The technology established in this study can be directly applied to produce functional gametes of endangered bitterlings using cryopreserved donor cells.

Genome and transcriptome analyses providing insight into the immune response of pearl oysters after allograft and xenograft transplantations.

The immune response after allograft or xenograft transplantation in the pearl oyster is a major factor that cause its nucleus rejection and death. To determine the mechanism underlying the immune response after allograft and xenograft transplantations in the pearl oyster Pinctada fucata martensii, we constructed two sets of transcriptomes of hemocytes at different times (6 and 12 h; 1, 3, 6, 12, and 30 d) after allograft and xenograft transplantations, in which the xenografted mantle tissue was from Pinctada maxima. The transcriptomic analysis reveals many genes are involved in the immune response to transplantation, such as transient receptor potential cation channel (TRP), calmodulin (CaM), DNA replication-related genes, and sugar and lipid metabolism-related genes. The expression of these identified genes was higher in the host pearl oyster transplanted with xenograft than that by allograft. The histological analysis of the pearl sac also confirmed that many hemocytes were still gathered around the transplanted nucleus, and no pearl sac was formed in the host pearl oysters at 30 d after xenograft transplantation. The genomic analysis indicated that pearl oysters evolved many copies of genes, such as TRP, CaM, and GST, to sense and cope with the immune response after transplantation. "Ribosome" and "Cytosolic DNA-sensing pathway" were specifically induced in the xenograft group, whereas "Notch signaling pathway" specifically responded to the allograft transplantation. These results can improve our understanding of the mechanism underlying the immune response of pearl oysters after allograft and xenograft transplantations.

Surgical management of long bone fractures in cats using cortical bone allografts preserved in honey.

This report describes comminuted fractures in 2 cats repaired by stabilization with cortical bone allografts that had been preserved in honey. Both cats exhibited appropriate post-operative weight-bearing. Preservation of bone in honey is a simple and widely available alternative for bone banking. This is the first report of a bone defect in cats repaired with bone preserved in honey.

Gestion chirurgicale de fractures osseuses longues chez les chats à l'aide d'une allogreffe de la corticale préservée dans du miel. Ce rapport décrit deux fractures comminutives réparées par la stabilisation à l'aide de l'allogreffe de la corticale qui avait été préservée dans du miel. Les deux chats ont manifesté une mise en charge postopératoire appropriée. La préservation de l'os dans du miel est une méthode de remplacement simple et facilement disponible comme ostéothèque. Il s'agit du premier rapport d'un défaut osseux chez des chats qui a été réparé à l'aide d'un os préservé dans du miel.(Traduit par Isabelle Vallières).

Corneal grafting for the treatment of full-thickness corneal defects in dogs: a review of 50 cases.

OBJECTIVE: To describe corneal grafting for the treatment of full-thickness corneal defects in dogs and to determine its effectiveness in preserving vision. METHODS: A review of the medical records of dogs that underwent corneal grafting following corneal perforations (≥3 mm) at the VTH-UAB from 2002 to 2012 was carried out. RESULTS: Fifty dogs of different breed, age and gender were included. Brachycephalic breeds were overrepresented (37/50;74%). All cases were unilateral, with euryblepharon being the most common concurrent ocular abnormality (20/50;40%). Full-thickness penetrating keratoplasties (FTPK) were performed in 21/50 eyes (42%) and lamellar keratoplasties (LK) in 29/50 eyes (58%). Frozen grafts (FroG) were used in 43/50 eyes (86%) and fresh homologous grafts (FreHoG) in 7/50 (14%). Of the former group, 26 were homologous (FroHoG:60%) and 17 heterologous (FroHeG:40%). A combination of topical medication (antibiotics, corticosteroids, cycloplegics, and 0.2% cyclosporine A) and systemic mycophenolate mofetil was administered. Median follow-up time was 200 days. Postsurgical complications included wound dehiscence (6/50;12%) and glaucoma (4/50;8%). Clinical signs of graft rejection were diagnosed as follows: FroHoG (13/26;50%), FroHeG (11/17;65%), FreHoG (4/7;57%), FTPK (12/21;57%), and LK (16/29;55%). Medical treatment successfully controlled graft rejection in 11/28 eyes (39%). Good anatomical outcome was achieved in 86% (43/50), of which 95% (41/43) were visual at last examination, with moderate opacification to complete transparency of the graft present in 48.2%. CONCLUSIONS: Corneal grafting is an effective surgical treatment for full-thickness corneal defects in dogs. If graft rejection is present, additional medical or surgical therapy may be necessary, achieving a highly satisfactory visual outcome.

Investigating the efficacy of amnion-derived compared with bone marrow-derived mesenchymal stromal cells in equine tendon and ligament injuries.

BACKGROUND AIMS: This is the first study to compare the treatment of horse tendon and ligament injuries with the use of mesenchymal stromal cells (MSCs) obtained from two different sources: amniotic membrane (AMSCs) and bone marrow (BM-MSCs). The objective was to prove the ability of AMSCs to exert beneficial effects in vivo. METHODS: Five million allogeneic frozen-thawed AMSCs or autologous fresh BM-MSCs were injected intralesionally in horses belonging to group A (51 horses) and group B (44 horses). The interval lesion/implantation was of 6-15 days for the AMSCs and 16-35 days for the BM-MSCs. Healing was assessed clinically and ultrasonographically. Follow-up was monitored for 2 further years from return to full work. RESULTS: No significant adverse effects after MSCs treatment were seen in any of the horses studied, independent of the type of stromal cell implanted. All animals belonging to group A resumed their activities between 4-5 months after treatment, whereas animals of group B resumed their activities after 4-12 months. The rate of re-injury in horses treated with AMSCs is lower (4.00%) compared with the average observed when horses were treated with BM-MSCs (23.08%). CONCLUSIONS: The possibility to inject allogeneic AMSCs in real time, before any ultrasonographic change occurs within the injured tendon and ligament, together with the higher plasticity and proliferative capacity of these cells compared with BM-MSCs, represents the main features of interest for this novel approach for the treatment of equine tendon diseases. An obvious active proliferative healing in the area injected with AMSCs makes these cells more effective than BM-MSCs.

A model for cryobanking female germplasm in Japanese quail (Coturnix japonica).

Cryopreservation and transplantation of ovarian tissue can be used for cryobanking female germplasm of avian species. Here we report a model of cryobanking ovarian tissue of Japanese quail. Ovaries of 1-wk-old quail chicks from the recessive white-breasted (WB) line were cryopreserved using a vitrification method. The cryopreserved tissue was warmed and transplanted orthotopically into ovariectomized, 1-wk-old female quail chicks from the homozygous wild-type QO line. At sexual maturation, recipients were mated with WB males and the plumage coloration was used as a marker to determine the origin of their ovaries. Seven of the 15 recipients laid eggs and produced donor-derived offspring, and 5 produced 100% donor-derived offspring. No difference was seen in egg production between recipients and untreated birds. The high efficiency of the vitrification and transplantation procedures in preserving and recovering quail ovarian tissue shows that this model can be used for maintaining commercial and experimental quail strains and may benefit other poultry species and endangered avian species.

The biomechanical and biological effect of supercooling on cortical bone allograft.

BACKGROUND: The need for a storage method capable of preserving the intrinsic properties of bones without using toxic substances has always been raised. Supercooling is a relatively recently introduced preservation method that meets this need. Supercooling refers to the phenomenon of liquid in which the temperature drops below its freezing point without solidifying or crystallizing. OBJECTIVES: The purpose of this study was to identify the preservation efficiency and applicability of the supercooling technique as a cortical bone allograft storage modality. METHODS: The biomechanical effects of various storage methods, including deep freezing, cryopreservation, lyophilization, glycerol preservation, and supercooling, were evaluated with the three-point banding test, axial compression test, and electron microscopy. Additionally, cortical bone allografts were applied to the radial bone defect in New Zealand White rabbits to determine the biological effects. The degree of bone union was assessed with postoperative clinical signs, radiography, micro-computed tomography, and biomechanical analysis. RESULTS: The biomechanical properties of cortical bone grafts preserved using glycerol and supercooling method were found to be comparable to those of normal bone while also significantly stronger than deep-frozen, cryopreserved, and lyophilized bone grafts. Preclinical research performed in rabbit radial defect models revealed that supercooled and glycerol-preserved bone allografts exhibited significantly better bone union than other groups. CONCLUSIONS: Considering the biomechanical and biological superiority, the supercooling technique could be one of the optimal preservation methods for cortical bone allografts. This study will form the basis for a novel application of supercooling as a bone material preservation technique.

Allograft theory: transmission of devil facial-tumour disease.

The Tasmanian devil, a large carnivorous Australian marsupial, is under threat from a widespread fatal disease in which a malignant oral-facial tumour obstructs the animal's ability to feed. Here we show that the chromosomes in these tumours have undergone a complex rearrangement that is identical for every animal studied. In light of this remarkable finding and of the known fighting behaviour of the devils, we propose that the disease is transmitted by allograft, whereby an infectious cell line is passed directly between the animals through bites they inflict on one another.

Allogeneic peripheral blood haematopoietic stem cell transplantation for the treatment of dogs with high-grade B-cell lymphoma.

Autologous peripheral blood haematopoietic stem cell transplantation (HCT) cures 33%-40% of dogs with high-grade B-cell lymphoma. We hypothesized, based on human allogeneic bone marrow transplantation literature, that transplanting dogs using canine donor leukocyte-matched CD34+ cells would lead to fewer relapses and increased cure rates. We retrospectively reviewed medical records of dogs diagnosed with high-grade B-cell lymphoma who received an identical allogeneic HCT. A total of 15 dogs transplanted at four facilities were identified. Five of fifteen dogs relapsed before transplant. The mean number of donor CD34+ cells/kg harvested and infused into recipient dogs was 8.0 × 106 /kg (range: 2.08 × 106 /kg-2.9 × 107 /kg). The median disease-free interval and overall survival of all dogs was 1095 days (range: 9-2920 days) and 1115 days (range: 9-2920 days), respectively. Two of five dogs, not in remission at transplant, died in the hospital. The median disease-free interval and overall survival of the remaining three dogs was 25 days (range: 15-250 days) and 1100 days (range: 66-1902 days), respectively. The median disease-free interval and overall survival of the 10 dogs who had not relapsed was 1235 days (range: 19-2920 days) and 1235 days (range: 19-2920 days), respectively. One dog died soon after discharge of presumed gastric-dilatation-volvulus. Eight of nine remaining dogs lived >4 yrs post-alloHCT, leading to a cure rate of 89%. Acute graft versus host disease was seen in three dogs. These results suggest that allogeneic HCT can cure ~50% more dogs than those treated with autologous HCT.

Novel Achilles tendon repair technique utilizing an allograft and hybrid external fixator in dogs.

Background: Achilles mechanism rupture is a surgical condition involving primary tenorrhaphy with various described means of surgical augmentation and bolstering. Aim: To report complications and outcomes with a novel Achilles repair technique in dogs using a superficial digital flexor tendon (SDFT) or deep digital flexor tendon (DDFT) allograft. Methods: Medical records were reviewed for dogs with chronic rupture or deterioration of the Achilles mechanism. Fibrous tissue was excised and either primary tenorrhaphy or reattachment of the tendon(s) to the calcaneus was performed. The surgical repair was supplemented by an SDFT or DDFT allograft, and postoperative immobilization was provided using a transarticular hybrid external skeletal fixator. Complications were classified as minor, major, or catastrophic, and function was classified as full, acceptable, or unacceptable, based on established guidelines. Results: Complications occurred with 6 out of 12 repairs, including 1 minor, 6 major, and 2 catastrophic complications. The two catastrophic complications were the recurrence of tarsal hyperflexion and lameness at 20 weeks and 18 months following surgery. Of the 12 surgeries performed, 2 resulted in full function, 8 with acceptable function, and 2 with unacceptable function at last follow-up 17-98 weeks postsurgery (mean = 45 weeks) for a success rate of 10/12 cases. Conclusion: The use of SDFT or DDFT allografts, coupled with an external fixator, can provide a moderate rate of full or acceptable functional outcomes and appears a viable treatment. However, complications were frequent and without a comparison group no conclusions can be drawn about the inferiority or superiority of this technique to other techniques for Achilles mechanism repair in dogs.

The efficacy of allogeneic mesenchymal precursor cells for the repair of an ovine tibial segmental defect.

INTRODUCTION: Synthetic void-fillers offer an alternative to autograft or allograft bone in the repair of segmental defects. However, the reparative process is delayed as only osteoconductive elements are present. The inclusion of pluripotential cells may resolve this limitation, and the use of allogeneic tissue provides the opportunity for an off-the-shelf remedy. The current study evaluated the utilisation of mesenchymal precursor cells (MPC) for the repair of an ovine critical-size tibial segmental defect. METHODS: Twenty-four, mature female sheep underwent surgery for the creation of a 3 cm tibial diaphyseal defect. In one group of 12 sheep the scaffold was used alone, and in the second group the scaffold was seeded with MPC. The defect was stabilised using a locking intramedullary nail and allowed to heal over a nine-month-period. Outcome assessments of healing included radiology of callus formation, computed tomography, assessment of new-bone volume, mechanical attributes, and histological evaluation of linear bone apposition rate and tissue response. RESULTS: The MPC-treated group displayed a significantly greater level of callus formation and rate of bone apposition in the defect. DISCUSSION: The incorporation of allogeneic MPC to a synthetic void filler stimulated early repair of critical-size diaphyseal segmental defects and holds potential as an off-the-shelf therapy for augmenting bone regeneration.

Fetal allogeneic umbilical cord cell transplantation improves motor function in spinal cord-injured rats.

AIM: The objective of this study was to investigate whether the transplantation of fetal umbilical cord tissue cells as a source of stem cells into the acutely injured spinal cord would produce some regenerations and/or functional recovery in a rat model of spinal cord injury. MATERIAL AND METHODS: Five pregnant albino Wistar rats of 12 days gestation were used for obtaining an umbilical cord cell graft. At the second stage of the experiment only Th8-Th9 laminectomy was performed in Group A animals while Group B animals underwent spinal cord hemitransection. The cultured fetal umbilical cord cells coated with Alginate Gel were placed into the lesion cavity immediately after surgery in Group C animals. Group D animals received only Alginate gel sponges into the injured area. All experiment groups were analyzed histologically and immunohistochemically (GFAP, Ki-67, and Pan cadherin) and for motor function after surgery. RESULTS: The umbilical cord cell transplanted animals showed a significant motor recovery compared to non-transplanted animals at 8 and 21 days after spinal cord injury (p=0.008). Significant GFAP and Ki-67 expressions were noted in transplanted animals (p=0.048) suggesting astroglial proliferation. CONCLUSION: Our findings support the possibility of some functional recovery after umbilical cord cell transplantation following spinal cord injury.

Improvement of the application of gonadal tissue allotransplantation in the in vitro conservation of chicken genetic lines.

In avian species, the surgical technique for ovarian allotransplantation has been developed for domestic chickens; however, not all genotypes can be effectively used as recipients. The aims of the present study were to ascertain donor/recipient combinations for production of offspring from frozen/thawed ovarian tissues. The development of the technique is important because domestic chicken offspring have only been produced from fresh (never frozen) ovarian and from frozen-thawed testicular tissues. Information obtained from evaluating genetic differences of intensively selected lines in which there was successful pairing was compared in the indigenous breeds. Results indicate donor/recipient combinations were created which could be effectively used for gonadal tissue allotransplantations. Gonadal tissues of Yellow, Speckled and Partridge-color Hungarian, Black and Speckled Transylvanian Naked Neck chicken breeds were allotransplanted into White Leghorn or Novogen White breeds for offspring production. The gonadal tissues of these indigenous breeds were cryopreserved using vitrification procedures. There was successful allografting of frozen/thawed gonadal tissues at a rate between 20 % and 100 % depending on the genotype and sex, and histological examination and microsatellite marker analysis provided evidence that the donor ovarian and testicular tissues had the capacity for producing gametes. The hens of Speckled Transylvanian Naked Neck/White Leghorn combination using frozen/thawed ovarian tissues were produced for progeny tests. Of these, 58 % produced eggs and 9.1 % produced donor-derived offspring, based on data for both feather color markers and genetic analysis.

Mesenchymal stem cells for treatment of musculoskeletal disease in horses: Relative merits of allogeneic versus autologous stem cells.

Mesenchymal stem cells (MSCs) are widely used for treatment of musculoskeletal diseases in horses, but there is ongoing debate regarding the relative safety and efficacy of allogeneic MSCs, compared with autologous equine MSCs. This review summarises the currently available published data regarding the therapeutic use of autologous and allogeneic MSCs in horses. Arguments that have been advanced against the use of allogeneic MSCs include higher risk of immunological reactions and shorter cell survival times following injection. Arguments favouring the use of allogeneic MSCs include the ability to bank cells and reduce the time to treatment, to collect MSCs from younger donor animals and the ability to manipulate banked cells prior to administration. In vitro studies and a limited set of experimental in vivo studies have indicated that adverse immunological reactions may occur when allogeneic MSCs are administered to horses. However, newer studies lack evidence of inflammatory reactions or adverse clinical responses when allogeneic MSCs are administered and compared with autologous MSCs. Thus, while the relative merits of allogeneic vs autologous MSCs for treatment of musculoskeletal injuries in horses have not been fully established, accumulating evidence from studies in horses suggests that allogeneic MSCs maybe a safe alternative to autologous MSCs. Large, properly designed, randomised trials in addition to careful immunological evaluation of short-term and long-term, local and systemic immune responses are needed to more fully resolve the issue.

The utility of DLA typing for transplantation medicine in canine models.

Transplantation medicine is used for the treatment of severe canine diseases, and the dog leukocyte antigen (DLA) is considered to be important in graft rejection. However, the utility of direct sequencing of both DLA classes I and II has not been assessed thoroughly. Eight healthy beagles with identified DLA genes were divided into two sets of four dogs, each including one donor and three recipients for skin transplantation. The following recipients were selected: one dog with a complete match, one with a haploidentical match, and one with a complete mismatch of the DLA gene with the donor. Full-thickness skin segments were obtained from each donor and transplanted to the recipients. A mixed lymphocyte reaction (MLR) assay was performed and analyzed by flow cytometry. Skin grafts of DLA haploidentical and mismatched pairs were grossly rejected within 14 days, whereas in fully matched DLA pairs, survival was as long as 21 days. Histopathological evaluation also showed moderate to severe lymphocytic infiltration and necrosis in DLA mismatched pairs. As seen in the MLR assay, the stimulation index of DLA mismatched pairs was significantly higher than that of fully matched DLA pairs in both sets (P<0.001). The allogeneic transplantation results suggested that it is possible to prolong transplant engraftment by completely matching the DLA genotype between the donor and recipient. Additionally, the MLR assay may be used as a simplified in vitro method to select donors.

Treatment of canine osteoarthritis with allogeneic platelet-rich plasma: review of five cases.

Background: Osteoarthritis (OA) is a major cause of chronic pain and lameness in dogs. Platelet-rich plasma (PRP) is a concentrate of growth and differentiation factors from the blood, which can be used in regenerative medicine strategies. Aim: The main aim of this study was to evaluate the effect of allogeneic PRP on the treatment of canine OA. Methods: Five dogs from several breeds, between 6 and 12 years old, and from both genders were studied. Clinical and imageological examinations diagnosed OA in the knee, tibiotarsal, elbow, and intercarpal joints. These dogs were refractory to medical therapy and to physical rehabilitation protocols that included shockwave therapy, laser therapy, electrostimulation, hydrotherapy, and diathermy.Animals were treated with allogeneic PRP obtained from the blood of the five dogs, which was processed in a pool. Echoguided intra-articular PRP injection was administered under sedation and after aseptic field preparation. Lameness at walk and trot (five grades) and pain (five scores) were evaluated before treatment and 30, 60, and 90 days post-treatment. Results: All animals presented improvements at 30 and 60 days in both parameters. Four dogs showed a decrease of three grades of lameness after 90 days and there was complete absence of lameness in 2 days. Pain was reduced from severe and moderate to mild in all the dogs after 30 days, and among them, three revealed no pain after 90 days. Conclusion: This study sheds light on the applicability and safety of a single administration of allogeneic PRP in osteoarthritic dogs.