Pharmacokinetics and tolerability of single-dose enteral cannabidiol and cannabidiolic acid rich hemp in horses (Equus caballus).

The pharmacokinetics and tolerability of cannabinoids and their metabolites were determined in eight horses after enteral administration of a commercial CBD/CBDA-rich hemp oil product. Each horse was administered 2 mg/kg or 8 mg/kg CBD/CBDA or no treatment in a randomized cross-over design. Serial serum samples collected over 48 h were analyzed by high performance liquid chromatography with tandem mass spectrometry. Plasma chemistry analysis was performed at 0 h and 24 h. Vital parameters, pedometry, and blinded mentation and gait evaluations were recorded at intervals up to 24 h. Manure production and gastrointestinal transit time were tracked for 48 h after oil administration. The median maximal concentration of CBD and CBDA were 5.2 and 36.95 ng/mL in the 2 mg/kg group, respectively; and 40.35 and 353.56 ng/mL in the 8 mg/kg group. The median half-life of elimination was not calculated for the 2 mg/kg CBD treatment due to lack of time points above the lower quantifiable limit beyond the Cmax while it was 7.75 h in the 8 mg/kg group. CBDA absorption was biphasic. Pharmacokinetic parameters for tetrahydrocannabinol, tetrahydrocannabinolic acid, cannabigerolic acid, and 7-carboxy cannabidiol are also reported. No significant differences in any of the measured tolerability parameters were demonstrated between treatment groups. Single-dose enteral administration of CBD/CBDA-rich hemp extract up to 8 mg/kg does not appear to produce neurologic, behavioral, or gastrointestinal effects in horses.

Antimicrobial prophylaxis is not indicated for horses undergoing general anaesthesia for elective orthopaedic MRI.

BACKGROUND: Post-anaesthetic fever is a known complication of general anaesthesia, however, its incidence in horses undergoing elective magnetic resonance imaging (MRI) is unknown. OBJECTIVE: To determine the incidence of post-anaesthetic fever in horses undergoing elective orthopaedic MRI and determine whether prophylactic antimicrobial therapy would be associated with a reduction in the incidence of post-anaesthetic fever. We hypothesised that prophylactic antimicrobials would be associated with a reduction in the incidence of post-anaesthetic fever. STUDY DESIGN: Retrospective cross-sectional study. METHODS: This retrospective study included 791 elective orthopaedic MRIs in systemically healthy horses between June 2006 and March 2020 that recovered from general anaesthesia and did not undergo surgery or intensive medical therapy soon after recovery. Potential factors associated with post-anaesthetic fever were evaluated using multivariable logistic regression. Case signalment, travel time, preanaesthetic haematology and fibrinogen abnormalities, use of prophylactic antimicrobials, peri-anaesthetic nonsteroidal anti-inflammatories, anaesthesia time and recovery time were all evaluated for association with post-anaesthetic fever. RESULTS: Of 791 MRI cases, 44 (5.6%) developed a post-anaesthetic fever. Horses that received prophylactic antimicrobials were [odds ratio (OR) 3.8, 95% confidence interval (CI) 1.98-7.46; p ≤ 0.001] more likely to develop a post-anaesthetic fever than those that did not receive antimicrobials. Young horses (1-4 years of age) were (OR 2.8, 95% CI 1.26-6.17; p = 0.01) more likely to develop fever compared with adult horses (≥5 years of age). MAIN LIMITATIONS: Limitations of this study pertain to retrospective analysis including nonrandomised case selection and incomplete data records. CONCLUSIONS: While fever may indicate infection, the majority of early post-anaesthetic fevers resolved before discharge from the hospital with no identified cause. The use of prophylactic antimicrobials to reduce the risk of post-anaesthetic fever for elective MRI is not supported by this study.

CONTEXTO: Febre é uma complicação comum após anestesia geral. Contudo, a incidência de febre em cavalos submetidos à ressonância magnética (RM) é desconhecida. OBJETIVO: Determinar a incidência de febre pós­anestésica em cavalos submetidos à RM devido à lesões ortopédicas e determinar se terapia antimicrobiana é necessária para reduzir a incidência de febre pós­anestésica. Nossa hipótese é que o uso de antimicrobianos é associado com a redução da incidência de febre pós­anestésica. DELINEAMENTO DO ESTUDO: Estudo retrospectivo transversal. METODOLOGIA: Esse estudo retrospectivo incluiu 791 equinos submetidos à RM por motivos ortopédicos, entre Junho de 2006 e Março de 2020, que recuperaram de anestesia geral, e não foram submetidos à cirurgia ou terapia intensa logo após a recuperação. Fatores que potencialmente poderiam ser associados com febre pós­anestésica foram avaliados utilizando regressão logística multivariada. Informações do paciente, como sexo e idade, tempo de viagem, anormalidades nos exames de sangue (hemograma e bioquímico) pré­anestésico, uso profilático de antimicrobianos, uso de anti­inflamatório não­esteroidal no período peri­anestésico, tempo de anestesia, e tempo de recuperação foram avaliados para possível associação com febre pós­anestésica. RESULTADOS: Dos 791 casos de RM, 44 (5.6%) desenvolveram febre pós­anestésica. Cavalos que receberam terapia antimicrobiana profilática foram (OR 3.8, 95% CI 1.98­7.46; p ≤ 0.001) vezes mais prováveis de desenvolverem febre pós­anestésica do que aqueles que não receberam antimicrobianos. Cavalos jovens (1­4 anos de idade) foram OR 2.8, 95% CI 1.26­6.17; p = 0.01) vezes mais prováveis de desenvolverem febre comparado com cavalos adultos (≥5 anos de idade). PRINCIPAIS LIMITAÇÕES: As limitações deste estudo são aquelas de uma análise retrospectiva, incluindo a seleção não randomizada dos pacientes e prontuários incompletos. CONCLUSÕES: Enquanto febre pode indicar a presença de infecção, a maioria das febres no período logo após anestesia se resolveram antes da alta do hospital e não tiveram nenhuma causa identificada. O uso profilático de antimicrobianos para reduzir a possível chance de febre pós­anestésica em casos de RM eletiva não é suportada por este estudo.

Equine Platelet-Rich Plasma.

Platelet-rich plasma (PRP) is an orthobiologic therapy composed of platelets, leukocytes, red blood cells, and plasma proteins. PRP has been used for 20 years, but progress determining efficacy has been slow. The definitions and classification of PRP are reviewed, and the use of PRP for tendon, ligament, and joint disease is discussed with a focus on findings of basic science and clinical studies, platelet activation, concurrent administration of nonsteroidal anti-inflammatory drugs, and treatment complications. Finally, the advantages of platelet lysates and freeze-dried platelets are discussed. The promising results of a PRP lysate optimized for antibiofilm and antimicrobial properties are introduced.

Three-dimensional models of the equine larynx can be used to perform traditional measures of arytenoid abduction and permit the positioning of modeled implants to demonstrate the anatomic feasibility of placing a rigid implant across the cricoarytenoid joint.

OBJECTIVE: To develop 3D models of larynges to compare arytenoid abduction measurements between specimens and models, and to investigate the anatomic feasibility of placing an implant across the cricoarytenoid joint (CAJ) with or without arthrotomy. SAMPLES: Cadaveric equine larynges (n = 9). PROCEDURES: Equine larynges underwent sequential CT scans in a neutral position and with 2 arytenoid treatments: bilateral arytenoid abduction (ABD) and bilateral arytenoid abduction after left cricoarytenoid joint arthrotomy (ARTH). Soft tissue, cartilage, and luminal volume 3-dimensional models were generated. Rima glottidis cross-sectional area (CSA) and left-to-right quotient (LRQ) angles were measured on laryngeal specimens and models. Arytenoid translation, articular contact area, and length of modeled implants placed across the CAJ were measured on models. Data were analyzed using paired t test or ANOVA and Tukey's post hoc test or non-parametric equivalents (P < .05). RESULTS: ARTH CSA was larger for laryngeal specimens than models (P = .0096). There was no difference in all other measures of CSA and LRQ angle between treatment groups or between specimens and models. There was no difference between ABD and ARTH groups for arytenoid cartilage translation, contact area, and implant length. The articular contact area was sufficient for modeled implant placement across the CAJ with a narrow range of implant lengths (17.59 mm to 23.87 mm) across larynges with or without arthrotomy. CLINICAL RELEVANCE: These results support further investigation of a CT-guided, minimally invasive surgical procedure. Future studies will evaluate the outcomes of the new procedure for technical precision, biomechanical stability, and post-operative success rates for horses with recurrent laryngeal neuropathy (RLN).

Pharmacokinetics of aminocaproic acid in thoroughbred horses.

The antifibrinolytic agent aminocaproic acid (ACA) is occasionally used prior to episodes of intense training in racehorses suffering from exercise-induced pulmonary hemorrhage. Although a previous study indicated that the drug is cleared rapidly in horses, some racetrack practitioners claim that recent adverse analytical findings for ACA in postrace samples were from ACA administrations 5-7 days before the race. The purpose of this study was to re-examine the pharmacokinetics of ACA in horses to address this apparent conundrum. Eight exercise-conditioned thoroughbred horses were administered 5 g of ACA IV, and blood and urine samples were collected at pre-determined time points prior to drug administration and for up to 168 h after dosing. Concentrations of ACA in the serum and urine samples were determined by LC-MS/MS. The pharmacokinetics of ACA in serum were best described by a three-compartment model with a terminal elimination half-life of 24.2 ± 2.9 h. After dosing, ACA was above the lower limit of detection (1 ng/mL for serum and 10 ng/mL for urine) in all serum and urine samples at all time points. In a similar manner, ACA was above the lower limit of quantification (LLOQ; 10 ng/mL for serum and 100 ng/mL for urine) in all serum and urine samples collected from all horses from 0.5 to 120 h post dosing. In six of the eight horses, ACA was above the LLOQ 168 h after dosing in serum and urine samples. LC-MS/MS methodology is the industry standard for testing of samples collected from racehorses with the purpose of controlling the use of medications and performance altering substances. The improved sensitivity of the analytical procedure used in the present study allowed the detection of a prolonged terminal elimination phase of ACA in horses that had not previously been described. Currently, most racing jurisdictions have not adopted a permitted concentration or threshold for ACA in postrace samples, and therefore veterinarians need to allow for an extended withdrawal time of a minimum of 11 days after the administration of ACA to racehorses to substantially reduce the risk of adverse analytical findings of ACA in postrace samples.

Kinematics of the equine distal sesamoid (navicular) bone of the thoracic limb.

OBJECTIVE: To quantify the translation and angular rotation of the distal sesamoid bone (DSB) using computed tomography (CT) and medical modeling software. SAMPLE: 30 thoracic limbs from equine cadavers. PROCEDURES: Partial (n = 12), full (8), and matched full and subsequently transected (10) thoracic limbs were collected. Bone volume CT images were acquired in three positions: extension (200° metacarpophalangeal angle), neutral (180°), and maximal flexion (110°). Mean translation and angular rotation of each DSB were recorded. Differences were determined with two-way ANOVA and post hoc Tukey's tests for pairwise comparisons; P value was set at < 0.05. RESULTS: Dorsal translation was significant during extension (1.4 ± 0.4 mm full limbs and 1.3 ± 0.2 mm partial limbs, P < 0.001). Distal translation was significant during extension (1.9 ± 0.4 mm full and 1.1 ± 0.4 mm partial) and flexion (5.4 ± 0.7 mm full and 6.22 ± 0.6 mm partial, P < 0.001). Rotation was significant (P < 0.001) about the mediolateral axis during extension (17.1° ± 1.4°) and flexion (2.6° ± 1.3°). Translation and rotation of the DSB were significantly different (P < 0.001) between full and partial limbs. CLINICAL RELEVANCE: This study provides the first quantification of translation and angular rotation of the DSB within the equine hoof. Partial limbs had significantly reduced movement compared to full limbs, suggesting that transection of flexor tendons alters distal thoracic limb kinematics. Further studies are required to determine if pathologic changes in the podotrochlear apparatus have an impact in clinical lameness outcomes.

Novel Methods to Mobilize, Isolate, and Expand Mesenchymal Stem Cells.

Numerous studies demonstrate the essential role of mesenchymal stem cells (MSCs) in the treatment of metabolic and inflammatory diseases, as these cells are known to modulate humoral and cellular immune responses. In this manuscript, we efficiently present two novel approaches to obtain MSCs from equine or human sources. In our first approach, we used electro-acupuncture as previously described by our group to mobilize MSCs into the peripheral blood of horses. For equine MSC collection, culture, and expansion, we used the Miltenyi Biotec CliniMACS Prodigy system of automated cell manufacturing. Using this system, we were able to generate appoximately 100 MSC colonies that exhibit surface marker expression of CD105 (92%), CD90 (85%), and CD73 (88%) within seven days of blood collection. Our second approach utilized the iPSC embryoid bodies from healthy or diabetic subjects where the iPSCs were cultured in standard media (endothelial + mesoderm basal media). After 21 days, the cells were FACS sorted and exhibited surface marker expression of CD105, CD90, and CD73. Both the equine cells and the human iPSC-derived MSCs were able to differentiate into adipogenic, osteogenic, and chondrogenic lineages. Both methods described simple and highly efficient methods to produce cells with surface markers phenotypically considered as MSCs and may, in the future, facilitate rapid production of MSCs with therapeutic potential.

Three-dimensional printed guides for screw placement in equine navicular bones.

OBJECTIVE: To determine the influence of a custom 3D-printed guide for placement of cortical bone screws in the equine navicular bone. STUDY DESIGN: Ex vivo study. SAMPLE POPULATION: Eight pairs of normal adult equine forelimbs. METHODS: A 3.5 × 55 mm cortical screw was placed in the longitudinal axis of each intact navicular bone. Screws were placed with a 3D-printed guide (3D) in one bone and with a traditional aiming device (AD) in the contralateral bone within each pair. Duration of surgery and the number of fluoroscopy images were compared between techniques. Screw placement was subjectively evaluated by gross examination and scored by three boarded veterinary surgeons. RESULTS: The use of a 3D-printed guide reduced the duration of surgery by 6.6 min (±1.5 min) compared to traditional screw placement (20.7 min ± 4.8 min, p < .01). Fewer peri-operative fluoroscopic images were obtained when the 3D guide was used (18 images ± 2.6 images vs. 40 images ± 5.1, p < .01). No difference was detected in navicular screw placement. CONCLUSION: The use of a 3D guide decreased the time required to place screws and the number of intraoperative images taken without affecting screw placement in intact navicular bones. CLINICAL RELEVANCE: 3D-printed guides can aid in the study, practice, and execution of surgical procedures reducing surgical time and radiation exposure throughout the operative period achieving similar results to those obtained with a conventional approach.

Endoscopic Laser Ablation of a Midline Cyst of the Colliculus Seminalis as a Treatment for Anejaculation in a Stallion.

An 18-year-old Appaloosa stallion presented with a history of ejaculatory dysfunction, which had recently progressed to an inability to ejaculate (anejaculation). Transrectal ultrasound evaluation revealed the presence of a prominent midline cyst of the colliculus seminalis, which was compressing the most terminal parts of the deferent ducts. Both ducts were enlarged and filled with hyperechoic content. The stallion was diagnosed with a complete occlusion of the deferent ducts because of the compression from the midline cyst of the colliculus seminalis. To date, there are no established treatments for this condition in stallions, although several procedures have been successfully used in men affected by similar problems. Therefore, we proposed performing one of these procedures-a laser ablation of the cyst to collapse it and to open the ejaculatory path. The stallion was placed under general anesthesia, and an endoscopic ultrasound-guided laser ablation of the cyst was performed. No immediate or long-term complications were observed. After the expulsion of accumulated material, the stallion produced normal semen and successfully impregnated mares during the next breeding season.

The effect of four different freezing conditions and time in frozen storage on the concentration of commonly measured growth factors and enzymes in equine platelet-rich plasma over six months.

BACKGROUND: Platelet-rich plasma (PRP) is a therapeutic biologic that is used for treatment of musculoskeletal pathologies in equine athletes. Due to the expense of PRP kits, and the volumes obtained, freezing aliquots for future dosing is common. Aliquots of PRP are also commonly frozen for later analysis of growth factor concentrations in in vitro research. A variety of freezing methods are used and storage duration until analysis is often not reported. The optimal frozen storage conditions and duration to maintain concentrations of commonly measured growth factors and enzymes in PRP are unknown. Our objectives were two-fold. First, to determine the effect of a single freeze-thaw cycle on PRP protein concentrations and establish their baseline levels. Second, to evaluate the effect of storage in -20 °C automatic defrost freezer, - 20 °C manual defrost freezer, - 80 °C manual defrost freezer, and liquid nitrogen for 1, 3, and 6 months on PRP protein concentrations, compared to the established baseline concentrations. RESULTS: Fold-change between fresh activated and snap frozen PRP were analyzed using paired t-test. A snap frozen-thaw cycle resulted in increased MMP-9 (p = 0.0021), and a small significant decrease in TGF-ß1 (p = 0.0162), while IGF-1 and PDGF-BB were unchanged compared to fresh activated PRP. Fold-change over time within storage method were analyzed using repeated measures ANOVA and Tukey post-hoc test. IGF-1 decreased in all conditions (p < 0.0001). At all time-points at -20 °C (p < 0.0001), and at 3 and 6 months at -80 °C (p < 0.0070), PDGF-BB decreased. TGF- ß1 was unchanged or increased after 6 months (p < 0.0085). MMP-9 decreased at 3-months at -20 °C, and at all times at -80 °C and in liquid nitrogen compared to snap frozen (p < 0.0001). CONCLUSIONS: The protein profile of equine frozen-stored PRP differs from fresh PRP. For clinical applications equine PRP can be stored at -80 °C for 1 month or in liquid nitrogen for 6 months to maintain PDGF-BB and TGF-ß1 concentration, but IGF-1 concentrations will be reduced. The storage temperature and duration should be reported in studies measuring protein concentrations in PRP. To accurately measure IGF-1 concentrations, PRP samples should be analyzed immediately.

Peripheral blood-derived mesenchymal stem cells demonstrate immunomodulatory potential for therapeutic use in horses.

Previously, we showed that mesenchymal stem cells (MSC) can be mobilized into peripheral blood using electroacupuncture (EA) at acupoints, LI-4, LI-11, GV-14, and GV-20. The purpose of this study was to determine whether EA-mobilized MSC could be harvested and expanded in vitro to be used as an autologous cell therapy in horses. Peripheral blood mononuclear cells (PBMC) isolated from young and aged lame horses (n = 29) showed a marked enrichment for MSCs. MSC were expanded in vitro (n = 25) and administered intravenously at a dose of 50 x 106 (n = 24). Treatment resulted in significant improvement in lameness as assessed by the American Association of Equine Practitioners (AAEP) lameness scale (n = 23). MSCs exhibited immunomodulatory function by inhibition of lymphocyte proliferation and induction of IL-10. Intradermal testing showed no immediate or delayed immune reactions to MSC (1 x 106 to 1 x 104). In this study, we demonstrated an efficient, safe and reproducible method to mobilize and expand, in vitro, MSCs in sufficiently high concentrations for therapeutic administration. We confirm the immunomodulatory function of these cells in vitro. This non-pharmacological and non-surgical strategy for stem cell harvest has a broad range of biomedical applications and represents an improved clinically translatable and economical cell source for humans.

Minimally Manipulated Bone Marrow Concentrate Compared with Microfracture Treatment of Full-Thickness Chondral Defects: A One-Year Study in an Equine Model.

BACKGROUND: Microfracture is commonly performed for cartilage repair but usually results in fibrocartilage. Microfracture augmented by autologous bone marrow concentrate (BMC) was previously shown to yield structurally superior cartilage repairs in an equine model compared with microfracture alone. The current study was performed to test the hypothesis that autologous BMC without concomitant microfracture improves cartilage repair compared with microfracture alone. METHODS: Autologous sternal bone marrow aspirate (BMA) was concentrated using a commercial system. Cells from BMC were evaluated for chondrogenic potential in vitro and in vivo. Bilateral full-thickness chondral defects (15-mm diameter) were created on the midlateral trochlear ridge in 8 horses. Paired defects were randomly assigned to treatment with BMC without concomitant microfracture, or to microfracture alone. The repairs were evaluated at 1 year by in vitro assessment, arthroscopy, morphological magnetic resonance imaging (MRI), quantitative T2-weighted and ultrashort echo time enhanced T2* (UTE-T2*) MRI mapping, and histological assessment. RESULTS: Culture-expanded but not freshly isolated cells from BMA and BMC underwent cartilage differentiation in vitro. In vivo, cartilage repairs in both groups were fibrous to fibrocartilaginous at 1 year of follow-up, with no differences observed between BMC and microfracture by arthroscopy, T2 and UTE-T2* MRI values, and histological assessment (p > 0.05). Morphological MRI showed subchondral bone changes not observed by arthroscopy and improved overall outcomes for the BMC repairs (p = 0.03). Differences in repair tissue UTE-T2* texture features were observed between the treatment groups (p < 0.05). CONCLUSIONS: When BMC was applied directly to critical-sized, full-thickness chondral defects in an equine model, the cartilage repair results were similar to those of microfracture. Our data suggest that, given the few mesenchymal stem cells in minimally manipulated BMC, other mechanisms such as paracrine, anti-inflammatory, or immunomodulatory effects may have been responsible for tissue regeneration in a previous study in which BMC was applied to microfractured repairs. While our conclusions are limited by small numbers, the better MRI outcomes for the BMC repairs may have been related to reduced surgical trauma to the subchondral bone. CLINICAL RELEVANCE: MRI provides important information on chondral defect subsurface repair organization and subchondral bone structure that is not well assessed by arthroscopy.

Two-Year Evaluation of Osteochondral Repair with a Novel Biphasic Graft Saturated in Bone Marrow in an Equine Model.

Objective To evaluate a biphasic cartilage repair device (CRD) for feasibility of arthroscopic implantation, safety, biocompatibility, and efficacy for long-term repair of large osteochondral defects. Methods The CRD was press-fit into defects (10 mm diameter, 10 mm deep) created in the femoral trochlea of 12 horses. In the contralateral limb, 10 mm diameter full-thickness chondral defects were treated with microfracture (MFX). Radiographs were obtained pre- and postoperatively, and at 4, 12, and 24 months. Repeat arthroscopy was performed at 4 and 12 months. Gross assessment, histology, mechanical testing, and magnetic resonance imaging (MRI) were performed at 24 months. Results The CRD was easily placed arthroscopically. There was no evidence of joint infection, inflammation, or degeneration. CRD-treated defects had significantly more sclerosis compared to MFX early ( P = 0.0006), but was not different at 24 months. CRD had better arthroscopic scores at 4 months compared to MFX ( P = 0.0069). At 24 months, there was no difference in repair tissue on histology or mechanical testing. Based on MRI, CRD repair tissue had less proteoglycan (deep P = 0.027, superficial P = 0.015) and less organized collagen (deep P = 0.028) compared to MFX. Cartilage surrounding MFX defects had more fissures compared to CRD. Conclusion The repair tissue formed after CRD treatment of a large osteochondral lesion is fibrocartilage similar to that formed in simple chondral defects treated with MFX. The CRD can be easily placed arthroscopically, is safe, and biocompatible for 24 months. The CRD results in improved early arthroscopic repair scores and may limit fissure formation in adjacent cartilage.

Angular Limb Deformities: Growth Retardation.

Angular limb deformities are common in foals; however, the importance of the deformity and if treatment is required depend on the degree of deformity relative to normal conformation for stage of growth, the breed and discipline expectations, age, and response to conservative therapies. This article addresses the importance of the foal conformation examination to determine which foals need surgical intervention to correct an angular deformity and when. Techniques for surgical growth retardation include the transphyseal staple, screw and wire transphyseal bridge, and transphyseal screw. Appropriate timing for intervention for each location and complications associated with each procedure are discussed.

Update on laryngeal disorders and treatment.

Laryngeal disorders are relatively common in the horse, and thorough diagnostic evaluation is essential to make an accurate definitive diagnosis and selection of appropriate treatment. The value of exercising endoscopy must not be overlooked, and the recent development of dynamic (overground) endoscopy is providing new insights into dynamic laryngeal lesions. The focus of this article will be on recently described disorders and treatments or modifications to existing treatments. It summarizes the numerous investigations attempting to perfect the laryngoplasty procedure for treatment of laryngeal hemiplegia. The newly described conditions, bilateral dynamic laryngeal collapse, and dynamically flaccid epiglottis will also be discussed.

Considerations for the use of platelet-rich plasma in orthopedics.

The use of platelet-rich plasma (PRP) is expanding to numerous medical fields, including orthopedic surgery and sports medicine. The popularity of this new treatment option has prompted a rapid increase in research endeavors; however, the differences in application technique and the composition of PRP have made it difficult to compare results or make any firm conclusions regarding efficacy. The purpose of this article is twofold. First, to recommend details that should be provided in basic science and clinical PRP studies to allow meaningful comparisons between studies which may lead to a better understanding of efficacy. Second, to provide an understanding of the different PRP preparations and their clinical relevance. There are biochemical rationales for the use of PRP because it addresses several aspects of the healing process, including cell proliferation and tissue matrix regeneration, inflammation, nociception, infection, and hemostasis, all of which will be addressed. Given the current understanding of the importance the composition of PRP plays in tissue regeneration, it is likely that our future understanding of PRP will dictate 'customizing' the PRP preparation to the specific pathology of interest. The potential complications following PRP use are minor, and thus it appears to be a safe treatment option with a variety of potentially beneficial effects to injured musculoskeletal tissues.

Optimization of leukocyte concentration in platelet-rich plasma for the treatment of tendinopathy.

BACKGROUND: Numerous methods are available for platelet-rich plasma (PRP) generation, but evidence defining the optimum composition is lacking. We hypothesized that leukocyte-reduced PRP would result in lower inflammatory cytokine expression compared with concentrated-leukocyte PRP and that maintaining the platelet:white blood cell (WBC) ratio would compensate for the effect of increased WBC concentration. METHODS: Blood and flexor digitorum superficialis tendons were collected from young adult horses. Three PRP groups were generated with the same platelet concentration but different WBC concentrations: intermediate-concentration standard PRP, leukocyte-reduced PRP, and concentrated-leukocyte PRP. An additional high-concentration PRP group was generated with the same WBC concentration as the concentrated-leukocyte PRP group and the same platelet:WBC ratio as the standard PRP group. The PRP groups were used as media for flexor digitorum superficialis tendon explants in culture for seventy-two hours with 10% plasma in Dulbecco modified Eagle medium (DMEM) serving as control. Tendon gene expression for collagen types I (COL1A1) and III (COL3A1), cartilage oligomeric matrix protein (COMP), matrix metalloproteinase (MMP-13), interleukin-1ß (IL-1ß), and tumor necrosis factor-α (TNF-α) was performed. RESULTS: The desired PRP groups were successfully generated. The expression of COMP, the COL1A1:COL3A1 ratio, and the expression of MMP-13 in flexor digitorum superficialis tendon explants was not different between PRP groups. The expression of COMP (p = 0.0027) and the COL1A1:COL3A1 ratio (p < 0.0001) were increased in the PRP groups as compared with the control group, and the expression of MMP-13 was decreased in the PRP groups as compared with the control group (p < 0.0001). The expression of IL-1ß was lowest in leukocyte-reduced PRP and highest in concentrated-leukocyte PRP (p = 0.0001). The leukocyte-reduced PRP group and the control group had the lowest TNF-α expression, whereas the high-concentration PRP and concentrated-leukocyte PRP groups had the highest expression (p = 0.0224). CONCLUSIONS: A high absolute WBC concentration in PRP contributes to the expression of inflammatory cytokines in flexor digitorum superficialis tendon explants, and maintenance of the platelet:WBC ratio is not able to counteract this effect. CLINICAL RELEVANCE: The optimum composition of PRP for the treatment of tendinopathy has not been directly investigated. Persistent inflammation results in inferior repair with scar tissue. The present study indicates that in an animal model, WBC in PRP contributes to inflammatory cytokine production. Therefore, leukocyte-reduced PRP may be the optimum preparation to stimulate superior healing without scar tissue formation.

The role of growth factors in cartilage repair.

BACKGROUND: Full-thickness chondral defects and early osteoarthritis continue to present major challenges for the patient and the orthopaedic surgeon as a result of the limited healing potential of articular cartilage. The use of bioactive growth factors is under consideration as a potential therapy to enhance healing of chondral injuries and modify the arthritic disease process. QUESTIONS/PURPOSES: We reviewed the role of growth factors in articular cartilage repair and identified specific growth factors and combinations of growth factors that have the capacity to improve cartilage regeneration. Additionally, we discuss the potential use of platelet-rich plasma, autologous-conditioned serum, and bone marrow concentrate preparations as methods of combined growth factor delivery. METHODS: A PubMed search was performed using key words cartilage or chondrocyte alone and in combination with growth factor. The search was open for original manuscripts and review papers and open for all dates. From these searches we selected manuscripts investigating the effects of growth factors on extracellular matrix synthesis and excluded those investigating molecular mechanisms of action. RESULTS: By modulating the local microenvironment, the anabolic and anticatabolic effects of a variety of growth factors have demonstrated potential in both in vitro and animal studies of cartilage injury and repair. Members of the transforming growth factor-ß superfamily, fibroblast growth factor family, insulin-like growth factor-I, and platelet-derived growth factor have all been investigated as possible treatment augments in the management of chondral injuries and early arthritis. CONCLUSIONS: The application of growth factors in the treatment of local cartilage defects as well as osteoarthritis appears promising; however, further research is needed at both the basic science and clinical levels before routine application.