Publications by authors named "Laurie Goodrich"

63 Publications

Intra-articular administration of antibiotics in horses: Justifications, risks, reconsideration of use and outcomes.

Equine Vet J 2021 Aug 30. Epub 2021 Aug 30.

Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado, USA.

Antibiotics have been injected intra-articularly by equine veterinarians for decades, either prophylactically when other drugs are administered for osteoarthritis or therapeutically to treat septic arthritis. This route of administration has also more recently gained attention in human orthopaedic clinical practice, particularly as an alternative to systemic antibiotic administration to treat infections following prosthetic arthroplasty. While the rationale for injecting antibiotics intra-articularly has been largely focused on achieving high local drug concentrations, there has been relatively little focus on pharmacokinetic parameters of antibiotics administered by this route, or on the potential for local toxicity. The increasing incidence of antibiotic resistance in veterinary and human medicine prompts reconsideration of off-label antibiotic usage and evaluation of evidence-based dosing strategies. The purpose of this review was to summarise the current literature describing intra-articular antibiotic usage, including specific studies where pharmacokinetics, potential safety and toxicity have been evaluated. This review will advance practitioners' understanding of the use of intra-articularly administered antibiotics, including the overall pros and cons of the approach.
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http://dx.doi.org/10.1111/evj.13502DOI Listing
August 2021

Evaluation of Intra-Articular Amikacin Administration in an Equine Non-inflammatory Joint Model to Identify Effective Bactericidal Concentrations While Minimizing Cytotoxicity.

Front Vet Sci 2021 21;8:676774. Epub 2021 May 21.

Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, United States.

Septic arthritis causes significant morbidity and mortality in veterinary and human clinical practice and is increasingly complicated by multidrug-resistant infections. Intra-articular (IA) antibiotic administration achieves high local drug concentrations but is considered off-label usage, and appropriate doses have not been defined. Using an equine joint model, we investigated the effects of amikacin injected at three different doses (500, 125, and 31.25 mg) on the immune and cartilage responses in tibiotarsal joints. Synovial fluid (SF) was sampled at multiple time points over 24 h, the cell counts determined, and amikacin concentrations measured by liquid chromatography-mass spectrometry. Cytokine concentrations and collagen degradation products in SF were measured by ELISA and multiplex immunoassays. The mean amikacin concentrations in SF were greater than or equal to the minimum inhibitory concentration (MIC) (0.004 mg/ml) for most common equine joint pathogens at all time points tested to 24 h for all three amikacin doses evaluated. The inflammatory cytokines tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) increased significantly in SF in the highest amikacin dose group, despite the fact that increases in SF cell counts were not observed. Similarly, the biomarkers of cartilage type II collagen cleavage (C2C and C12C) were increased in SF following amikacin injection. Mechanistically, we further demonstrated using studies that chondrocytes and synoviocytes killed by exposure to amikacin underwent apoptotic cell death and were phagocytosed by macrophages in a non-inflammatory process resembling efferocytosis. Neutrophils and T cells were susceptible to amikacin cytotoxicity at clinically relevant doses, which may result in blunting of cellular inflammatory responses in SF and account for the lack of increase in total nucleated cell counts following amikacin injection. In summary, decisions on whether to inject cytotoxic antibiotics such as aminoglycosides intra-articularly and what doses to use should take into account the potential harm that antibiotics may cause and consider lower doses than those previously reported in equine practice.
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http://dx.doi.org/10.3389/fvets.2021.676774DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8175670PMC
May 2021

Impact of Three Different Serum Sources on Functional Properties of Equine Mesenchymal Stromal Cells.

Front Vet Sci 2021 30;8:634064. Epub 2021 Apr 30.

Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, United States.

Culture and expansion of equine mesenchymal stromal cells (MSCs) are routinely performed using fetal bovine serum (FBS) as a source of growth factors, nutrients, and extracellular matrix proteins. However, the desire to minimize introduction of xenogeneic bovine proteins or pathogens and to standardize cellular products intended for clinical application has driven evaluation of alternatives to FBS. Replacement of FBS in culture for several days before administration has been proposed to reduce antigenicity and potentially prolong survival after injection. However, the functional consequences of MSC culture in different serum types have not been fully evaluated. The objective of this study was to compare the immunomodulatory and antibacterial properties of MSCs cultured in three serum sources: FBS or autologous or allogeneic equine serum. We hypothesized that continuous culture in FBS would generate MSCs with improved functionality compared to equine serum and that there would not be important differences between MSCs cultured in autologous vs. allogeneic equine serum. To address these questions, MSCs from three healthy donor horses were expanded in medium with FBS and then switched to culture in FBS or autologous or allogeneic equine serum for 72 h. The impact of this 72-h culture period in different sera on cell viability, cell doubling time, cell morphology, bactericidal capability, chondrogenic differentiation, and production of cytokines and antimicrobial peptides was assessed. Altering serum source did not affect cell viability or morphology. However, cells cultured in FBS had shorter cell doubling times and secreted more interleukin 4 (IL-4), IL-5, IL-17, RANTES, granulocyte-macrophage colony-stimulating factor, fibroblast growth factor 2, eotaxin, and antimicrobial peptide cathelicidin/LL-37 than cells cultured in either source of equine serum. Cells cultured in FBS also exhibited greater spontaneous bactericidal activity. Notably, significant differences in any of these parameters were not observed when autologous vs. allogeneic equine serum was used for cell culture. Chondrogenic differentiation was not different between different serum sources. These results indicate that MSC culture in FBS will generate more functional cells based on a number of parameters and that the theoretical risks of FBS use in MSC culture should be weighed against the loss of MSC function likely to be incurred from culture in equine serum.
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http://dx.doi.org/10.3389/fvets.2021.634064DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8119767PMC
April 2021

USE OF INTRACARPAL INTERLEUKIN RECEPTOR ANTAGONIST PROTEIN (IRAP) AND HYALURONIC ACID IN A MULTIMODAL THERAPEUTIC REGIME FOR OSTEOARTHRITIS IN AN ASIAN ELEPHANT ().

J Zoo Wildl Med 2021 Apr;52(1):401-405

Department of Clinical Sciences and Orthopedic Research Center, College of Veterinary Medicine and Biomedical Sciences, Fort Collins, CO 80523, USA.

An approximately 41-yr-old female Asian elephant ( experiencing forelimb stiffness and decreased range of motion was diagnosed with bilateral carpal osteoarthritis (OA). Standing sedation combined with local anesthesia was used to deliver ultrasound-guided carpal articular injections of an autologous conditioned serum product, interleukin receptor antagonist protein, combined with hyaluronic acid. Within 2 mo of completing therapy, improved range and speed of motion were evident. Reduced inflammation was suggested by decreased carpal articular prostaglandin E2 levels. Subjectively improved clinical signs lasted approximately 5-6 mo, at which point carpal articular injections were repeated. Joint inflammatory markers were useful in gauging response to treatment and may provide guidance in the diagnostic and therapeutic approach to elephant OA. On the basis of the positive response noted, interarticular autologous therapy combined with hyaluronic acid should be considered for carpal OA in elephants.
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http://dx.doi.org/10.1638/2019-0113DOI Listing
April 2021

Toll-like receptor activation of equine mesenchymal stromal cells to enhance antibacterial activity and immunomodulatory cytokine secretion.

Vet Surg 2021 May 2;50(4):858-871. Epub 2021 Apr 2.

Department of Clinical Sciences, College of Veterinary Medicine, Colorado State University, Colorado, USA.

Objective: To evaluate effects of Toll-like and nucleotide-binding oligomerization domain (NOD)-like receptor (TLR, NLR) ligand stimulation of equine mesenchymal stromal cells (MSCs) on antibacterial and immunomodulatory properties in vitro.

Study Design: Controlled laboratory study.

Sample Population: Equine bone-marrow-derived MSCs (three horses).

Methods: MSCs were stimulated with TLR (polyinosinic:polycytidylic acid [pIC] and lipopolysaccharide [LPS]) and NLR agonists (γ-d-Glu-mDAP [IE-DAP]) for 2 h, and plated at 1 × 10 cells/well 24 h. MSC-conditioned media (MSC-CM) were collected and assessed for antimicrobial peptide cathelicidin/LL-37 production, bactericidal action against multidrug-resistant planktonic and biofilm Staphylococcus aureus and neutrophil phagocytosis. Bacterial growth was measured by plating bacteria and counting viable colonies, reading culture absorbance, and live-dead staining with confocal microscopy imaging. Following initial comparison of activating stimuli, TLR3-agonist pIC protocols (cell density during activation and plating, culture time, %serum) were further optimized for bactericidal activity and secretion of interleukin-8 (IL-8), monocyte-chemoattractant-protein (MCP-1), and cathelicidin/LL37.

Results: MSCs stimulation with pIC (p = .004) and IE-DAP (p = .03) promoted increased bactericidal activity, evidenced by reduced viable planktonic colony counts. PIC stimulation (2 × 10 cells/ml, 2 h, 10 μg/ml) further suppressed biofilm formation (p = .001), enhanced neutrophil bacterial phagocytosis (p = .009), increased MCP-1 secretion (p < .0001), and enhanced cathelicidin/LL-37 production, which was apparent when serum concentration in media was reduced to 1% (p = .01) and 2.5% (p = .05).

Conclusion: TLR-3 pIC MSCs activation was most effective to enhance antibacterial and cytokine responses, which were affected by serum reduction.

Clinical Significance: In vitro TLR-3 activation of equine MSCs tested here may be a strategy to improve antibacterial properties of MSCs to treat antibiotic-resistant infections.
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http://dx.doi.org/10.1111/vsu.13628DOI Listing
May 2021

Quantitative Evaluation of Equine Articular Cartilage Using Cationic Contrast-Enhanced Computed Tomography.

Cartilage 2021 04 2;12(2):211-221. Epub 2018 Dec 2.

Boston University, Boston, MA, USA.

Objective: To investigate the diffusion trajectory of a cationic contrast medium (CA4+) into equine articular cartilage, and to assess normal and degenerative equine articular cartilage using cationic contrast-enhanced computed tomography (CECT).

Design: In the first experiment (Exp1), equine osteochondral specimens were serially imaged with cationic CECT to establish the diffusion time constant and time to reach equilibrium in healthy articular cartilage. In a separate experiment (Exp2), articular cartilage defects were created on the femoral trochlea (defect joint) in a juvenile horse, while the opposite joint was a sham-operated control. After 7 weeks, osteochondral biopsies were collected throughout the articular surfaces of both joints. Biopsies were analyzed for cationic CECT attenuation, glycosaminoglycan (GAG) content, mechanical stiffness (E), and histology. Imaging, biochemical and mechanical data were compared between defect and control joints.

Results: Exp1: The mean diffusion time constant was longer for medial condyle cartilage (3.05 ± 0.1 hours) than lateral condyle cartilage (1.54 ± 0.3 hours, = 0.04). Exp2: Cationic CECT attenuation was lower in the defect joint than the control joint ( = 0.005) and also varied by anatomic location ( = 0.045). Mean cationic CECT attenuation from the lateral trochlear ridge was lower in the defect joint than in the control joint (2223 ± 329 HU and 2667 ± 540 HU, respectively; = 0.02). Cationic CECT attenuation was strongly correlated with both GAG (ρ = 0.79, < 0.0001) and E (ρ = 0.61, < 0.0001).

Conclusions: The equilibration time of CA4+ into equine articular cartilage is affected by tissue volume. Quantitative cationic CECT imaging reflects the biochemical, biomechanical and histological state of normal and degenerative equine articular cartilage.
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http://dx.doi.org/10.1177/1947603518812562DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7970376PMC
April 2021

Susceptibility of canine chondrocytes and synoviocytes to antibiotic cytotoxicity in vitro.

Vet Surg 2021 Apr 19;50(3):650-658. Epub 2021 Feb 19.

Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Colorado.

Objective: To evaluate relative cytotoxicity of antibiotics to normal canine joint tissues in vitro.

Study Design: Experimental in vitro study.

Sample Population: Chondrocytes and synoviocytes (three dogs); cartilage explants (three dogs); six dogs total.

Methods: Chondrocytes and synoviocytes from normal femoropatellar joints of three dogs were plated on 24-well plates (50 000 cells/cm , triplicate, 48 hours) and exposed to antibiotics (ampicillin sulbactam, vancomycin, cefazolin, ceftazidime, amikacin, enrofloxacin; 0.39-25 mg/mL, 24 hours). Viability was assessed by using trypan blue dye exclusion. Antibiotic concentrations at which 50% cell death occurred (half-maximal inhibitory concentration) were determined to rank antibiotics for relative cytotoxicity. Occurrence of caspase-3 expression after antibiotic exposure was assessed as an indication of apoptosis induction. Cartilage explants from three different dogs were minced and exposed to antibiotics (amikacin, ceftazidime, cefazolin, enrofloxacin; 5 mg/mL, 72 hours). Live/dead staining was performed, and fluorescence was visualized by using confocal microscopy. Percentage of live vs dead cells was quantitated.

Results: Viability of chondrocytes and synoviocytes decreased with increasing antibiotic concentrations. Half-maximal inhibitory concentrations were determined for synoviocytes (vancomycin 13.77, ampicillin sulbactam 3.07, amikacin 2.26, ceftazidime 1.62, cefazolin 1.48, enrofloxacin 1.25 mg/mL) and chondrocytes (vancomycin 8.65, ampicillin sulbactam 8.63, ceftazidime 3.16, amikacin 2.74, cefazolin 1.67, enrofloxacin 0.78 mg/mL). Caspase-3 expression was upregulated, providing evidence that apoptotic pathways were active in cell death.

Conclusion: Half-maximal inhibitory concentration data provided evidence of lower toxicity of vancomycin and ampicillin sulbactam to joint tissues in vitro.

Clinical Significance: These results provide evidence to justify future in vitro work with osteoarthritic joint tissues and in vivo clinical trials to evaluate safety and efficacy of intra-articular antibiotics to treat dogs with septic arthritis.
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http://dx.doi.org/10.1111/vsu.13591DOI Listing
April 2021

Cationic contrast-enhanced computed tomography distinguishes between reparative, degenerative, and healthy equine articular cartilage.

J Orthop Res 2021 Aug 2;39(8):1647-1657. Epub 2020 Nov 2.

Orthopaedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Colorado State University, Fort Collins, Colorado, USA.

Cationic contrast-enhanced computed tomography (CECT) is a quantitative imaging technique that characterizes articular cartilage, though its efficacy in differentiating repair tissue from other disease states is undetermined. We hypothesized that cationic CECT attenuation will distinguish between reparative, degenerative, and healthy equine articular cartilage and will reflect biochemical, mechanical, and histologic properties. Chondral defects were created in vivo on equine femoropatellar joint surfaces. Within defects, calcified cartilage was retained (Repair 1) or removed (Repair 2). At sacrifice, plugs were collected from within defects, and at locations bordering (adjacent site) and remote to defects along with site-matched controls. Articular cartilage was analyzed via CECT using CA4+ to assess glycosaminoglycan (GAG) content, compressive modulus (E ), and International Cartilage Repair Society (ICRS) II histologic score. Comparisons of variables were made between sites using mixed model analysis and between variables with correlations. Cationic CECT attenuation was significantly lower in Repair 1 (1478 ± 333 Hounsfield units [HUs]), Repair 2 (1229 ± 191 HUs), and adjacent (2139 ± 336 HUs) sites when compared with site-matched controls (2587 ± 298, 2505 ± 184, and 2563 ± 538 HUs, respectively; all p < .0001). Cationic CECT attenuation was significantly higher at remote sites (2928 ± 420 HUs) compared with Repair 1, Repair 2, and adjacent sites (all p < .0001). Cationic CECT attenuation correlated with ICRS II score (r = .79), GAG (r = .76), and E (r = .71; all p < .0001). Cationic CECT distinguishes between reparative, degenerative, and healthy articular cartilage and highly correlates with biochemical, mechanical, and histological tissue properties.
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http://dx.doi.org/10.1002/jor.24894DOI Listing
August 2021

One health in regenerative medicine: report on the second Havemeyer symposium on regenerative medicine in horses.

Regen Med 2020 06 18;15(6):1775-1787. Epub 2020 Aug 18.

Department of Clinical Sciences & Services, The Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Herts. AL9 7TA, UK.

Regenerative medicine is commonly used in human and equine athletes. Potential therapies include culture expanded stem cells, stromal vascular fraction of adipose tissue, platelet-rich plasma, bone marrow concentrate, or autologous conditioned serum. The purpose of this manuscript is to disseminate findings from a workshop on the development of translational regenerative medicine in the equine field. Five themes emerged: stem cell characterization and tenogenic differentiation; interactions between mesenchymal stem cells, other cells and the environment; scaffolds and cell packaging; blood- and bone marrow-based regenerative medicines; clinical use of regenerative therapies. Evidence gained through the use of regenerative medicine applications in the horse should continue to translate to the human patient, bringing novel regenerative therapies to both humans and horses.
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http://dx.doi.org/10.2217/rme-2019-0143DOI Listing
June 2020

The platelet-rich plasma and mesenchymal stem cell milieu: A review of therapeutic effects on bone healing.

J Orthop Res 2020 12 17;38(12):2539-2550. Epub 2020 Jul 17.

Department of Clinical Sciences, Orthopaedic Research Center, Translational Medicine Institute, College of Veterinary Medicine, Colorado State University, Fort Collins, Colorado.

Platelet-rich plasma is autologous plasma that contains concentrated platelets compared to whole blood. It is relatively inexpensive to produce, can be easily isolated from whole blood, and can be administered while the patient is in the operating room. Further, because platelet-rich plasma is an autologous therapy, there is minimal risk for adverse reactions to the patient. Platelet-rich plasma has been used to promote bone regeneration due to its abundance of concentrated growth factors that are essential to wound healing. In this review, we summarize the methods for producing platelet-rich plasma and the history of its use in bone regeneration. We also summarize the growth factor profiles derived from platelet-rich plasma, with emphasis on those factors that play a direct role in promoting bone repair within the local fracture environment. In addition, we discuss the potential advantages of combining platelet-rich plasma with mesenchymal stem cells, a multipotent cell type often obtained from bone marrow or fat, to improve craniofacial and long bone regeneration. We detail what is currently known about how platelet-rich plasma influences mesenchymal stem cells in vitro, and then highlight the clinical outcomes of administering platelet-rich plasma and mesenchymal stem cells as a combination therapy to promote bone regeneration in vivo.
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http://dx.doi.org/10.1002/jor.24786DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8354210PMC
December 2020

Use of in vitro assays to identify antibiotics that are cytotoxic to normal equine chondrocytes and synovial cells.

Equine Vet J 2021 May 3;53(3):579-589. Epub 2020 Jul 3.

Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.

Background: Intra-articular (IA) antibiotic usage is prevalent in equine practice. However, recent emergence of antimicrobial resistance prompts re-evaluation of antibiotic selection, particularly when used prophylactically. Furthermore, many commonly used antibiotics exert direct cytotoxicity to equine cells, and appropriate IA doses have not been defined.

Objectives: To screen antibiotics in vitro as an initial assessment of cytotoxicity against normal equine joint cells in monolayer culture and explant tissues.

Study Design: In vitro experimental study.

Methods: Chondrocytes and synovial cells were harvested from three horses and plated on 24-well plates (100 000 cells/wells in triplicate) for 48 hours prior to addition of antibiotics. Joint cells were exposed to antibiotics (n = 15) at various doses (25-0.39 mg/mL in complete DMEM media) for 24 hours and viability was assessed by trypan blue dye exclusion. The half maximal inhibitory concentration (IC50) was determined for each antibiotic. Cartilage explants were obtained from 3 horses, minced and exposed to antibiotics (n = 5) for 72 hours. Live/dead staining was performed, and fluorescence was visualised using Olympus IX83 spinning disk confocal microscope. Percentage of live vs dead cells was quantified.

Results: Antibiotics from different antimicrobial classes expressed dose-dependent but variable cytotoxicity to equine joint cells in vitro. Aminoglycosides and doxycycline had the lowest IC50 (most toxic). Ampicillin sulbactam, imipenem, tobramycin, ceftiofur sodium and amoxicillin had IC50 > 25 mg/mL for at least one cell line, representing potentially less cytotoxic alternatives.

Main Limitations: Further studies are necessary to extrapolate these in vitro data results to the in vivo joint environment.

Conclusions: Targeted IA antibiotic therapy would involve selection of the safest antibiotics (highest IC50) with efficacy based on bacterial culture/sensitivity. Antimicrobial selection and evidence-based dosing may minimise damage to native articular cartilage and synovial cells and development of antimicrobial resistance when IA antibiotics are used in equine practice.
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http://dx.doi.org/10.1111/evj.13314DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7738387PMC
May 2021

What Is Your Diagnosis?

J Am Vet Med Assoc 2020 Jun;256(11):1205-1207

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http://dx.doi.org/10.2460/javma.256.11.1205DOI Listing
June 2020

Adult ovine chondrocytes in expansion culture adopt progenitor cell properties that are favorable for cartilage tissue engineering.

J Orthop Res 2020 09 6;38(9):1996-2005. Epub 2020 Apr 6.

Department of Clinical Sciences, Orthopaedic Reserch Center, C. Wayne McIlwraith Translational Medicine Institute, Fort Collins, Colorado.

Human chondrocytes in expansion culture can become progenitor-like in their ability to proliferate extensively and secrete neocartilage in chondrogenic culture. Sheep are used as a large animal model for cartilage tissue engineering, although for testing progenitor-like chondrocytes it is important that ovine chondrocytes resemble human in the ability to adopt progenitor properties. Here, we investigate whether ovine chondrocytes can adopt progenitor properties as indicated by rapid proliferation in a colony-forming fashion, and high levels of neocartilage secretion in chondrogenic culture. In conditions known to promote expansion of mesenchymal stromal cells, ovine chondrocytes proliferated through approximately 12 population doublings in 10 days. Time-lapse imaging indicated rapid proliferation in a colony-forming pattern. Expanded ovine chondrocytes that were seeded into agarose and cultured in chondrogenic medium accumulated neocartilage over 2 weeks, to a greater extent than primary chondrocytes. These data confirm that ovine chondrocytes resemble human chondrocytes in their ability to acquire progenitor properties that are important for cartilage tissue engineering. Given the broad interest in using progenitor cells to heal connective tissues, next we compared proliferation and trilineage differentiation of ovine chondrocytes, meniscus cells, and tenocytes. Meniscus cells and tenocytes experienced more than 13 population doublings in 10 days. In chondrogenic culture, cartilage matrix accumulation, and gene expression were largely similar among the cell types. All cell types resisted osteogenesis, while expanded tenocytes and meniscal cells were capable of adipogenesis. While ovine connective tissue cells demonstrated limited lineage plasticity, these data support the potential to promote certain progenitor properties with expansion.
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http://dx.doi.org/10.1002/jor.24671DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8442064PMC
September 2020

Amikacin induces rapid dose-dependent apoptotic cell death in equine chondrocytes and synovial cells in vitro.

Equine Vet J 2020 Sep 21;52(5):715-724. Epub 2020 Feb 21.

Translational Medicine Institute, College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO, USA.

Background: Equine veterinarians frequently inject aminoglycoside antibiotics intra-articularly, either to treat septic arthritis or for prophylaxis with other medications when injecting joints for osteoarthritis. Although aminoglycosides have been demonstrated to be toxic to equine mesenchymal stem cells (MSC), their effects on resident joint cells have not been previously investigated. Moreover, safe and effective intra-articular doses have not been defined.

Objectives: To determine effects of concentration, duration of exposure, pH and the presence of synovial fluid on the cytotoxic effects of amikacin on equine chondrocytes, synoviocytes and bone marrow- and adipose-derived MSC.

Study Design: In vitro experimental study.

Methods: Four cell types were harvested from three donor horses and plated in triplicate wells for 48 hours prior to the addition of amikacin. The effects of amikacin on cell viability were assessed for different exposure times, concentrations and with pH buffered or unbuffered in media, as well as in the presence of synovial fluid. Cell metabolism/viability was assessed by colorimetric MTT assay. Cell proliferation was assessed by live cell imaging. Cell viability was assessed using trypan blue and dimeric cyanine nucleic acid stain (yoyo-1). To determine the mechanism of cell death, apoptosis was evaluated using Annexin V and 7AAD staining with flow cytometric quantification. Induction of apoptotic cell death pathways was assessed using caspase-3 expression.

Results: Amikacin is cytotoxic to equine joint cells and MSC in a rapid, dose-dependent, pH-independent manner, which occurs primarily by apoptosis. Amikacin cytotoxicity was not mitigated by the addition of synovial fluid in vitro.

Main Limitations: Further studies are necessary to determine whether these in vitro results predict joint injury in live animal models.

Conclusions: Amikacin at clinically applied doses induces rapid, pronounced cell death of equine joint cells. These findings suggest that amikacin doses currently used intra-articularly should be reconsidered pending in vivo joint titration studies.
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http://dx.doi.org/10.1111/evj.13243DOI Listing
September 2020

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

Equine Vet J 2020 Sep 19;52(5):654-663. Epub 2020 Feb 19.

C. Wayne McIlwraith Translational Medicine Institute, Colorado State University College of Veterinary Medicine, Fort Collins, CO, USA.

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.
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http://dx.doi.org/10.1111/evj.13233DOI Listing
September 2020

Single and repeated intra-articular injections in the tarsocrural joint with allogeneic and autologous equine bone marrow-derived mesenchymal stem cells are safe, but did not reduce acute inflammation in an experimental interleukin-1β model of synovitis.

Equine Vet J 2020 Jul 14;52(4):601-612. Epub 2020 Feb 14.

Orthopedic Research Center, C. Wayne McIlwraith Translational Medicine Institute, Department of Clinical Sciences, College of Veterinary Medicine, Colorado State University, Fort Collins, Colorado, USA.

Background: Allogeneic and autologous bone marrow-derived mesenchymal stem cells (BMDMSCs) have been administered in equine joints for their anti-inflammatory effects. However, allogeneic BMDMSC offer multiple clinical and practical advantages. Therefore, it is important to determine the relative effectiveness of allogeneic vs autologous BMDMSCs.

Objectives: The objective of the study was to compare the inflamed joint response to autologous vs allogeneic BMDMSCs injections, and to determine if either treatment generated an anti-inflammatory effect.

Study Design: Randomised controlled study.

Method: Bone marrow was harvested from eight horses. Autologous BMDMSCs and pooled allogeneic BMDMSCs were culture expanded, cryopreserved and thawed immediately prior to administration. Ten million autologous BMDMSCs were administered with 75 ng rIL-1β into one tarsocrural joint and the contralateral tarsocrural joint received allogeneic BMDMSC plus 75 ng rIL-1β. Repeat injections were performed with the same treatment administered into the same joint. Four additional horses received 75 ng rIL-1β alone in a single tarsocrural joint. Clinical parameters (lameness, joint circumference and joint effusion) and synovial fluid parameters, including nucleated cell count (NCC), differential cell count, total protein (TP), prostaglandin E (PGE ) and C-reactive protein (CRP), were measured at baseline, 6, 12, 24, 72, 168 and 336 hours post-injection.

Results: No difference was detected between autologous and allogeneic treatment groups with respect to subjective lameness, joint effusion, joint circumference, NCC, TP, differential cell count, CRP or PGE . Neither autologous nor allogeneic treatments resulted in an improvement in clinical or cytological parameters over that elicited by rIL-1β alone.

Main Limitations: A single dose of rIL-1β was evaluated and resulted in a severe synovitis which may have been too severe to observe a BMDMSC-mediated effect.

Conclusions: This study revealed that allogeneic and autologous BMDMSCs resulted in an equivalent clinical and cytological response. Allogeneic and autologous BMDMSCs were equally ineffective in reducing the inflammatory response from acute rIL-1β-induced joint inflammation in horses.
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http://dx.doi.org/10.1111/evj.13222DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7283005PMC
July 2020

Can Extracorporeal Shockwave Promote Osteogenesis of Equine Bone Marrow-Derived Mesenchymal Stem Cells In Vitro

Stem Cells Dev 2020 01 17;29(2):110-118. Epub 2019 Dec 17.

Orthopedic Research Center at the C. Wayne McIlwraith Translational Medicine Institute, Department of Clinical Sciences, College of Veterinary Medicine, Colorado State University, Fort Collins, Colorado.

Both bone marrow-derived mesenchymal stem cells (BMDMSCs) and extracorporeal shockwave (ESW) have shown promise for enhancing fracture repair. If exposure of BMDMSCs to ESW enhances osteogenic differentiation, these therapies may be combined in vivo or used as a method for preconditioning BMDMSCs. The objective of this study was to determine the effect of ESW on the osteogenic ability of equine BMDMSCs. We hypothesized that ESW would promote osteogenesis evidenced by increased gene expression, alkaline phosphatase (ALPL) expression, slide morphologic score, and protein expression. BMDMSCs were evaluated from six horses. BMDMSCs were culture expanded to passage 3, dissociated, then placed in conical tubes. Treatment cells ("shocked") were exposed to 500 pulses at 0.16 mJ/mm energy. Cells were then reseeded and grown in either growth medium or osteogenic medium. Cellular proliferation and trilineage potential were determined. Cellular morphology was scored and cells were harvested at 1, 3, 7, 14, and 21 days for rtPCR gene expression of osteogenic markers [osteonectin (), osteocalcin (), ALPL, collagen type 3 (), and runt-related transcription factor 2 ()]. Media supernatants were evaluated for secretion of BMP-2, VEGF, TGFβ, and PGE and cellular lysates were evaluated for ALPL production. There was no difference between the proliferative ability of shocked cells versus unshocked cells in either growth medium or osteogenic medium. ALPL production was greater in shocked cells maintained in osteogenic medium versus unshocked cells in osteogenic medium at day 3 ( < 0.005). Independent of media type, ESW caused a decrease in VEGF and TGFβ production at day 3. No significant increases in gene expression were identified by rtPCR. Exposure of BMDMSCs to ESW does not result in negative effects. An initial significant increase in ALPL was detected but no persistent osteogenic effect was observed with cell expansion.
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http://dx.doi.org/10.1089/scd.2019.0202DOI Listing
January 2020

Long-term outcome after stifle arthroscopy in 82 Western performance horses (2003-2010).

Vet Surg 2019 Aug 14;48(6):956-965. Epub 2019 Jun 14.

Department of Clinical Sciences, Colorado State University, Fort Collins, Colorado.

Objective: To report the outcome of horses engaged in Western performance disciplines after stifle arthroscopy and identify prognostic factors for return to performance.

Study Design: Retrospective case series.

Sample Population: Eighty-two Western performance horses undergoing stifle arthroscopy.

Methods: Medical records were reviewed for horses involved in athletic performance/training for various Western performance disciplines and undergoing arthroscopy for lameness localized to the stifle. Follow-up was obtained ≥2 years postoperatively by telephone interviews with the owners. Preoperative and intraoperative findings as well as postoperative treatment were analyzed for their association with return to athletic performance as the primary outcome of interest.

Results: The most common disciplines represented were cutting (n = 38), Western pleasure (n = 13), and reining (n = 13). Approximately 40% (32/82) of horses returned to intended use after surgery. Increased age, higher degree of lameness, longer duration of lameness, and the presence of partial-thickness cartilage lesions decreased the odds of returning to athletic performance. Postoperative therapies (intra-articular: stem cells, corticosteroids, interleukin-1 receptor antagonist protein, hyaluronic acid/polysulfated glycosaminoglycans; systemic: nonsteroid anti-inflammatory drugs, hyaluronic acid/polysulfated glycosaminoglycans, oral joint supplements) did not affect the odds of returning to intended use.

Conclusion: Less than half of the Western performance horses that underwent stifle arthroscopy returned to intended use. Older age, longer duration of lameness, and presence of partial-thickness cartilage lesions affected the odds of a horse returning to intended use. Postoperative therapies did not affect the outcome in this population.

Clinical Significance: The prognosis of Western performance horses undergoing stifle arthroscopy is as guarded as that previously reported in horses of other disciplines.
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http://dx.doi.org/10.1111/vsu.13241DOI Listing
August 2019

Contrast-Enhanced Computed Tomography Scoring System for Distinguishing Early Osteoarthritis Disease States: A Feasibility Study.

J Orthop Res 2019 10 21;37(10):2138-2148. Epub 2019 Jun 21.

Department of Biomedical Engineering, Boston University, 590 Commonwealth Avenue, Boston, Massachusetts, 02215.

Early detection of osteoarthritis (OA) remains a diagnostic challenge owing to insensitive diagnostic techniques currently available. Herein a new semiquantitative scoring system, based upon contrast-enhanced computed tomographic (CECT) imaging, is described for further refinement of early OA disease staging. Trochlear ridge cartilage defects were surgically created in the femoropatellar joint of an adult horse (ACUC approved protocols). Seven weeks post-surgery, CECT imaging was performed on a clinical scanner after intra-articular injection of a cationic iodinated contrast agent, CA4+, into both injured and control femoropatellar joint compartments. The femoral cartilage surface was densely biopsied, and specimens were assessed for visual (Outerbridge score), functional (equilibrium compressive modulus), and biochemical (glycosaminoglycan content) measures of cartilage quality. Cartilage CECT attenuation was compared with cartilage quality measures using receiver operating characteristic curve analysis to establish attenuation thresholds for distinguishing among cartilage quality levels. CECT imaging identifies macroscopically damaged cartilage regions and in morphologically identical tissue provides moderately sensitive and specific semiquantitative segregation of cartilage quality based upon CECT attenuation, reflecting both glycosaminoglycan content and compressive stiffness of cartilage area under the curve (AUC = 0.83 [95% confidence interval [CI]: 0.72-0.93] for distinguishing poor quality and AUC = 0.76 [95% CI: 0.65-0.90] for distinguishing healthy quality cartilage). A semiquantitative 6-point scoring system-the Osteoarthritis Attenuation and Morphological Assessment (OAMA) score-is proposed as a tool for assessing cartilage quality from CECT images. The OAMA scoring system expands the current disease staging capability of early OA by inclusion of morphological, biochemical, and biomechanical assessments. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2138-2148, 2019.
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http://dx.doi.org/10.1002/jor.24382DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6739126PMC
October 2019

Usefulness of caudomedial-craniolateral oblique radiographic views for the diagnosis of injury to the origin of the cranial cruciate ligament in two horses.

J Am Vet Med Assoc 2019 Feb;254(4):508-511

CASE DESCRIPTION A 12-year-old mixed-breed mare (horse 1) and 6-year-old Friesian gelding (horse 2) were examined for chronic lameness associated with the stifle joint. CLINICAL FINDINGS Lameness examination revealed effusion of the right (horse 1) or left (horse 2) femoropatellar and medial femorotibial joints and grade 3/5 (horse 1) or 4/5 (horse 2) lameness. A diagnosis of cranial cruciate ligament (CCL) injury with associated mineralization and avulsion (horse 1) or mineralization alone (horse 2) was facilitated in both horses with a caudomedial-craniolateral oblique radiographic view obtained 45° medial to the caudocranial line, which highlighted the origin of the ligament on the caudoaxial aspect of the lateral femoral condyle within the intercondylar fossa. These lesions were subsequently confirmed via CT. TREATMENT AND OUTCOME Arthroscopy of the medial and lateral femorotibial joints was performed for horse 1 and revealed the osseous fragment associated with the CCL, but the fragment could not be removed. Horse 2 was euthanized while anesthetized following CT owing to the poor prognosis. CONCLUSIONS AND CLINICAL RELEVANCE Radiography is typically the first imaging modality attempted for horses with CCL injury, particularly outside the hospital setting. A 45° caudomedial-craniolateral oblique radiographic view may aid in diagnosis of CCL injury when avulsion or mineralization is present. Although this view is not commonly included in the typical radiographic series for imaging of the stifle joint in horses, it should be considered when CCL injury is suspected.
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http://dx.doi.org/10.2460/javma.254.4.508DOI Listing
February 2019

Genetic modification of scAAV-equine-BMP-2 transduced bone-marrow-derived mesenchymal stem cells before and after cryopreservation: An "off-the-shelf" option for fracture repair.

J Orthop Res 2019 06 21;37(6):1310-1317. Epub 2019 Feb 21.

Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, Fort Collins, CO 80523, USA.

Optimizing the environment of complex bone healing and improving treatment of catastrophic bone fractures and segmental bone defects remains an unmet clinical need both human and equine veterinary medical orthopaedics. The objective of this study was to determine whether scAAV-equine-BMP-2 transduced cells would induce osteogenesis in equine bone marrow derived mesenchymal stem cells (BMDMSCs) in vitro, and if these cells could be cryopreserved in an effort to osteogenically prime them as an "off-the-shelf" gene therapeutic approach for fracture repair. Our study found that transgene expression is altered by cell expansion, as would be expected by a transduction resulting in episomal transgene expression, and that osteoinductive levels could still be achieved 5 days after recovery, and protein expression would continue up to 14 days after transduction. This is the first evidence that cryopreservation of genetically modified BMDMSCs would not alter the osteoinductive potential or clinical use of allogeneic donor cells in cases of equine fracture repair. © 2018 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1310-1317, 2019.
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http://dx.doi.org/10.1002/jor.24209DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8366205PMC
June 2019

Photopolymerizable Injectable Cartilage Mimetic Hydrogel for the Treatment of Focal Chondral Lesions: A Proof of Concept Study in a Rabbit Animal Model.

Am J Sports Med 2019 01 27;47(1):212-221. Epub 2018 Nov 27.

Department of Clinical Sciences and Orthopaedic Research Center, Colorado State University, Fort Collins, Colorado, USA.

Background: In this study, we investigate the in vitro and in vivo chondrogenic capacity of a novel photopolymerizable cartilage mimetic hydrogel, enhanced with extracellular matrix analogs, for cartilage regeneration.

Purpose: To (1) determine whether mesenchymal stem cells (MSCs) embedded in a novel cartilage mimetic hydrogel support in vitro chondrogenesis, (2) demonstrate that the proposed hydrogel can be delivered in situ in a critical chondral defect in a rabbit model, and (3) determine whether the hydrogel with or without MSCs supports in vivo chondrogenesis in a critical chondral defect.

Study Design: Controlled laboratory study.

Methods: Rabbit bone marrow-derived MSCs were isolated, expanded, encapsulated in the hydrogel, and cultured in chondrogenic differentiation medium for 9 weeks. Compressive modulus was evaluated at day 1 and at weeks 3, 6, and 9. Chondrogenic differentiation was investigated via quantitative polymerase reaction, safranin-O staining, and immunofluorescence. In vivo, a 3 mm-wide × 2-mm-deep chondral defect was created bilaterally on the knee trochlea of 10 rabbits. Each animal had 1 defect randomly assigned to be treated with hydrogel with or without MSCs, and the contralateral knee was left untreated. Hence, each rabbit served as its own matched control. Three groups were established: group A, hydrogel (n = 5); group B, hydrogel with MSCs (n = 5); and group C, control (n = 10). Repair tissue was evaluated at 6 months after intervention.

Results: In vitro, chondrogenesis and the degradable behavior of the hydrogel by MSCs were confirmed. In vivo, the hydrogel could be delivered intraoperatively in a sterile manner. Overall, the hydrogel group had the highest scores on the modified O'Driscoll scoring system (group A, 17.4 ± 4.7; group B, 13 ± 3; group C, 16.7 ± 2.9) ( P = .11) and showed higher safranin-O staining (group A, 49.4% ± 20%; group B, 25.8% ± 16.4%; group C, 36.9% ± 25.2%) ( P = .27), although significance was not detected for either parameter.

Conclusion: This study provides the first evidence of the ability to photopolymerize this novel hydrogel in situ and assess its ability to provide chondrogenic cues for cartilage repair in a small animal model. In vitro chondrogenesis was evident when MSCs were encapsulated in the hydrogel.

Clinical Relevance: Cartilage mimetic hydrogel may offer a tissue engineering approach for the treatment of osteochondral lesions.
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http://dx.doi.org/10.1177/0363546518808012DOI Listing
January 2019

Use of a locking compression plate for equine proximal interphalangeal joint arthrodesis: 29 cases (2008-2014).

J Am Vet Med Assoc 2018 Dec;253(11):1460-1466

OBJECTIVE To describe clinical use of a locking compression plate (LCP) for proximal interphalangeal joint (PIPJ) arthrodesis in horses and compare outcomes for horses that underwent the procedure as treatment for fracture of the middle phalanx (P2) versus other causes. DESIGN Retrospective case series. ANIMALS 29 client-owned horses. PROCEDURES Medical records of 2 veterinary teaching hospitals from 2008 through 2014 were reviewed to identify horses that underwent PIPJ arthrodesis of 1 limb. Signalment, surgical, and outcome-related variables were recorded. Owners were contacted from 1 to 6 years after surgery to determine rehabilitation time, current use of the horse, and overall owner satisfaction with the procedure. Success was determined on the basis of owner satisfaction and outcome for intended use. Variables of interest were compared statistically between horses that underwent surgery for P2 fracture versus other reasons. RESULTS 14 horses underwent surgery for treatment of P2 fracture, and 15 had surgery because of osteoarthritis, subluxation, or osteochondrosis. Median convalescent time after surgery (with no riding or unrestricted exercise) was 7 months. Four horses were euthanized; of 23 known alive at follow-up, 22 were not lame, and 18 had returned to their intended use (8 and 10 at higher and lower owner-reported levels of work, respectively). Horses undergoing arthrodesis for reasons other than fracture were significantly more likely to return to their previous level of work. Twenty-two of 24 owners contacted indicated satisfaction with the procedure. CONCLUSIONS AND CLINICAL RELEVANCE Surgical arthrodesis of the PIPJ was successful in most horses of the study population. Various nuances of the system for fracture repair need to be understood prior to its use.
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http://dx.doi.org/10.2460/javma.253.11.1460DOI Listing
December 2018

Proceedings of the signature series symposium "cellular therapies for orthopaedics and musculoskeletal disease proven and unproven therapies-promise, facts and fantasy," international society for cellular therapies, montreal, canada, may 2, 2018.

Cytotherapy 2018 11 10;20(11):1381-1400. Epub 2018 Oct 10.

Department of Orthopedic Surgery and Biomedical Engineering Cleveland Clinic, Cleveland, Ohio, USA. Electronic address:

The Signature Series Symposium "Cellular Therapies for Orthopaedics and Musculoskeletal Disease Proven and Unproven Therapies-Promise, Facts and Fantasy" was held as a pre-meeting of the 26 International Society for Cellular Therapy (ISCT) annual congress in Montreal, Canada, May 2, 2018. This was the first ISCT program that was entirely dedicated to the advancement of cell-based therapies for musculoskeletal diseases. Cellular therapies in musculoskeletal medicine are a source of great promise and opportunity. They are also the source of public controversy, confusion and misinformation. Patients, clinicians, scientists, industry and government share a commitment to clear communication and responsible development of the field. Therefore, this symposium convened thought leaders from around the world in a forum designed to catalyze communication and collaboration to bring the greatest possible innovation and value to patients with musculoskeletal conditions.
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http://dx.doi.org/10.1016/j.jcyt.2018.09.001DOI Listing
November 2018

Optimizing Clinical Use of Biologics in Orthopaedic Surgery: Consensus Recommendations From the 2018 AAOS/NIH U-13 Conference.

J Am Acad Orthop Surg 2019 Jan;27(2):e50-e63

Concern that misinformation from direct-to-consumer marketing of largely unproven "biologic" treatments such as platelet-rich plasma and cell-based therapies may erode the public trust and the responsible investment needed to bring legitimate biological therapies to patients have resulted in calls to action from professional organizations and governing bodies. In response to substantial patient demand for biologic treatment of orthopaedic conditions, the American Academy of Orthopaedic Surgeons convened a collaborative symposium and established a consensus framework for improving and accelerating the clinical evaluation, use, and optimization of biologic therapies for musculoskeletal diseases. The economic and disease burden of musculoskeletal conditions is high. Of the various conditions discussed, knee osteoarthritis was identified as a "serious condition" associated with substantial and progressive morbidity and emerged as the condition with the most urgent need for clinical trial development. It was also recognized that stem cells have unique characteristics that are not met by minimally manipulated mixed cell preparations. The work group recommended that minimally manipulated cell products be referred to as cell therapy and that the untested and uncharacterized nature of these treatments be clearly communicated within the profession, to patients, and to the public. Minimum standards for product characterization and clinical research should also be followed. A framework for developing clinical trials related to knee OA was agreed upon. In addition to recommendations for development of high-quality multicenter clinical trials, another important recommendation was that physicians and institutions offering biologic therapies commit to establishing high-quality patient registries and biorepository-linked registries that can be used for postmarket surveillance and quality assessments.
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http://dx.doi.org/10.5435/JAAOS-D-18-00305DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6314629PMC
January 2019

Induction of Synovitis Using Interleukin-1 Beta: Are There Differences in the Response of Middle Carpal Joint Compared to the Tibiotarsal Joint?

Front Vet Sci 2018 31;5:208. Epub 2018 Aug 31.

Department of Clinical Sciences, Orthopaedic Research Center, Colorado State University, Fort Collins, CO, United States.

The effects of recombinant interleukin-1β (rIL-1β) have been described for the middle carpal joint (MCJ). However, we are unaware of any studies that have described the cytological response of the tibiotarsal joint (TTJ) to rIL-1β or compared the clinical and cytological responses of the MCJ to the TTJ following the administration of intra-articular rIL-1β. Such information is critical for researchers planning to use rIL-1β to create acute synovitis models in horses. To compare the clinical and cytological responses of the MCJ to the TTJ following administration of intra-articular rIL-1β. Twelve horses were used for the study. Eight horses received 75 ng of rIL-1β into the MCJ and four horses received 75 ng of rIL-1β into the TTJ. Clinical and cytological outcome parameters including lameness, joint circumference, joint effusion score, total nucleated cell count, cellular differentials, C-reactive protein, and prostaglandin-E2 concentrations were determined at baseline and multiple post-treatment time points over a 336 h period (2 weeks). Recombinant IL-1β administered into the TTJ resulted in a significantly greater respiratory rate at 24 h and heart rate at 12 h when compared to rIL-1β administered into the MCJ. In addition, the TTJ had a significantly greater increase in joint circumference at 24 post-injection hour (PIH) and subjective effusion grade at 24 PIH and 336 PIH. The MCJ had significantly higher total protein concentration at 6 PIH, and a significantly higher NCC at 24 and 72 PIH when compared to the TTJ. Conversely, the TTJ had significantly higher neutrophilic infiltration than the MCJ at 6 PIH and 168 PIH. This study establishes that the same intra-articular dose of rIL-1 β elicits significantly different clinical and cytological responses in the MCJ compared to the TTJ in the equine model of intra-articular synovitis. In addition, clinical and cytological evidence of synovitis may persist up to or >1 week following intra-articular administration of rIL-1 β.
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http://dx.doi.org/10.3389/fvets.2018.00208DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6127273PMC
August 2018

Pancarpal and partial carpal arthrodesis with 3 locking compression plates in 6 horses.

Vet Surg 2018 Jul;47(5):692-704

Department of Clinical Studies, New Bolton Center, University of Pennsylvania, Kennett Square, Pennsylvania.

Objective: To report the outcome of horses after pancarpal or partial carpal arthrodesis with 3 locking compression plates (LCP).

Study Design: Case series.

Animals: Six horses ranging in age from 8 months to 16 years and weighing 227-580 kg with severe carpal pathology including acute fractures, chronic osteoarthritis, and chronic angular limb deformity.

Methods: Pancarpal or partial carpal arthrodesis was performed with 3 LCP. Autologous cancellous bone grafts were used in 5 of 6 cases to facilitate joint arthrodesis.

Results: External coaptation was maintained for 4 to 6 weeks after surgery. Radiographic follow-up was available in all 6 cases, all of which reached arthrodesis and pasture soundness by 4-5 months postoperatively. One case required implant removal at 6 months because of implant exposure through the skin but returned to pasture soundness after removal.

Conclusion: Carpal instability due to acute fractures or chronic disease was successfully stabilized with 3 short LCP, leading to pasture soundness in all 6 horses.

Clinical Significance: The use of 3 short LCP should be considered as a strategy to facilitate pancarpal or partial carpal arthrodesis by providing superior stability without placement of implants in the diaphysis of the radius and third metacarpus.
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http://dx.doi.org/10.1111/vsu.12916DOI Listing
July 2018

Culture Conditions that Support Expansion and Chondrogenesis of Middle-Aged Rat Mesenchymal Stem Cells.

Cartilage 2020 07 28;11(3):364-373. Epub 2018 Jul 28.

The Feinstein Institute for Medical Research, North Shore-LIJ Health System, Manhasset, NY, USA.

Objective: Rats are an early preclinical model for cartilage tissue engineering, and a practical species for investigating the effects of aging. However, rats may be a poor aging model for mesenchymal stem cells (MSCs) based on laboratory reports of a severe decline in chondrogenesis beyond young adulthood. Such testing has not been conducted with MSCs seeded in a scaffold, which can improve the propensity of MSCs to undergo chondrogenesis. Therefore, the objective of this study was to evaluate chondrogenesis of middle-aged rat MSCs encapsulated in agarose.

Design: MSCs from 14- to 15-month-old rats were expanded, seeded into agarose, and cultured in chondrogenic medium with or without 5% serum for 15 days. Samples were evaluated for cell viability and cartilaginous extracellular matrix (ECM) accumulation. Experiments were repeated using MSCs from 6-week-old rats.

Results: During expansion, middle-aged rat MSCs demonstrated a diminishing proliferation rate that was improved ~2-fold in part by transient exposure to chondrogenic medium. In agarose culture in defined medium, middle-aged rat MSCs accumulated ECM to a much greater extent than negative controls. Serum supplementation improved cell survival ~2-fold, and increased ECM accumulation ~3-fold. Histological analysis indicated that defined medium supported chondrogenesis in a subset of cells, while serum-supplementation increased the frequency of chondrogenic cells. In contrast, young rat MSCs experienced robust chondrogenesis in defined medium that was not improved with serum-supplementation.

Conclusions: These data demonstrate a previously-unreported propensity of middle-aged rat MSCs to undergo chondrogenesis, and the potential of serum to enhance chondrogenesis of aging MSCs.
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http://dx.doi.org/10.1177/1947603518790047DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7298599PMC
July 2020

The challenges of promoting osteogenesis in segmental bone defects and osteoporosis.

J Orthop Res 2018 06 6;36(6):1559-1572. Epub 2018 Mar 6.

Orthopaedic Research Center, College of Veterinary Medicine, Colorado State University, 1678 Campus Delivery, Fort Collins, Colorado, 80523-1678.

Conventional clinical management of complex bone healing scenarios continues to result in 5-10% of fractures forming non-unions. Additionally, the aging population and prevalence of osteoporosis-related fractures necessitate the further exploration of novel ways to augment osteogenesis in this special population. This review focuses on the current clinical modalities available, and the ongoing clinical and pre-clinical research to promote osteogenesis in segmental bone defects, delayed unions, and osteoporosis. In summary, animal models of fracture repair are often small animals as historically significant large animal models, like the dog, continue to gain favor as companion animals. Small rodents have well-documented limitations in comparing to fracture repair in humans, and few similarities exist. Study design, number of studies, and availability of funding continue to limit large animal studies. Osteoinduction with rhBMP-2 results in robust bone formation, although long-term quality is scrutinized due to poor bone mineral quality. PTH 1-34 is the only FDA approved osteo-anabolic treatment to prevent osteoporotic fractures. Limited to 2 years of clinical use, PTH 1-34 has further been plagued by dose-related ambiguities and inconsistent results when applied to pathologic fractures in systematic human clinical studies. There is limited animal data of PTH 1-34 applied locally to bone defects. Gene therapy continues to gain popularity among researchers to augment bone healing. Non-integrating viral vectors and targeted apoptosis of genetically modified therapeutic cells is an ongoing area of research. Finally, progenitor cell therapies and the content variation of patient-side treatments (e.g., PRP and BMAC) are being studied. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:1559-1572, 2018.
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http://dx.doi.org/10.1002/jor.23845DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8354209PMC
June 2018

Use of Platelet-Rich Plasma Immediately After an Injury Did Not Improve Ligament Healing, and Increasing Platelet Concentrations Was Detrimental in an In Vivo Animal Model.

Am J Sports Med 2018 03 6;46(3):702-712. Epub 2017 Dec 6.

Steadman Philippon Research Institute, Vail, Colorado, USA.

Background: Limited information in basic science and clinical trials exists to determine if ligament healing may be accelerated with the use of biological adjuvants, such as platelet-rich plasma (PRP). However, there has been widespread acceptance of PRP for use in clinical practice, despite an inadequate understanding of its biological mechanism of action.

Purpose: To determine whether a single dose of PRP could accelerate ligament healing and correspondingly improve histological characteristics and biomechanical properties when injected immediately postoperatively into the injured medial collateral ligament (MCL) of New Zealand White rabbits.

Study Design: Controlled laboratory study.

Methods: Eighty skeletally mature New Zealand White rabbits (160 knees) were used. The MCL was torn midbody to simulate a grade 3 tear. After an acute injury of the MCL, the administration of autologous PRP at 3 different platelet concentrations (0 million/uL, platelet-poor plasma [PPP]; 0.6 million/uL, 2 times the baseline [2× PRP]; and 1.2 million/uL, 4 times the baseline [4× PRP]) was performed and compared with a saline injection control in the contralateral knee. Histological analysis and a biomechanical endpoint characterization were utilized to assess ligamentous healing and compare it to a sham surgery group.

Results: The PPP ( P = .001) and 4× PRP ( P = .002) groups had a significantly lower collagen subscore than the sham surgery group. No other differences were observed among the treatment groups, including the vascularity subscore and overall ligament tissue maturity index score. Compared with saline-injected contralateral knees, the maximum load for PPP and 2× PRP was not significantly different ( P = .788 and .325, respectively). The maximum load and stiffness for knees treated with 4× PRP were significantly less than for the saline-treated contralateral knees ( P = .006 and .001, respectively).

Conclusion: One single dose of PPP or 2× PRP at the time of injury did not improve ligament healing. In addition, 4× PRP negatively affected ligament strength and histological characteristics at 6 weeks after the injury.

Clinical Relevance: The current practice of treating knee ligament injuries with PRP may not improve healing at low doses of PRP. The decreased mechanical properties and histological appearance of the torn MCL suggest that high doses of PRP decrease the quality of repair tissue. Further in vivo studies are necessary to determine the dosing and timing of PRP administration after a ligament injury before the widespread use of PRP to treat ligament injuries is recommended.
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http://dx.doi.org/10.1177/0363546517741135DOI Listing
March 2018
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