Mechanistic insights into cancer cell killing through interaction of phosphodiesterase 3A and schlafen family member 12.

Cytotoxic molecules can kill cancer cells by disrupting critical cellular processes or by inducing novel activities. 6-(4-(Diethylamino)-3-nitrophenyl)-5-methyl-4,5-dihydropyridazin-3(2H)-one (DNMDP) is a small molecule that kills cancer cells by generation of novel activity. DNMDP induces complex formation between phosphodiesterase 3A (PDE3A) and schlafen family member 12 (SLFN12) and specifically kills cancer cells expressing elevated levels of these two proteins. Here, we examined the characteristics and covariates of the cancer cell response to DNMDP. On average, the sensitivity of human cancer cell lines to DNMDP is correlated with PDE3A expression levels. However, DNMDP could also bind the related protein, PDE3B, and PDE3B supported DNMDP sensitivity in the absence of PDE3A expression. Although inhibition of PDE3A catalytic activity did not account for DNMDP sensitivity, we found that expression of the catalytic domain of PDE3A in cancer cells lacking PDE3A is sufficient to confer sensitivity to DNMDP, and substitutions in the PDE3A active site abolish compound binding. Moreover, a genome-wide CRISPR screen identified the aryl hydrocarbon receptor-interacting protein (AIP), a co-chaperone protein, as required for response to DNMDP. We determined that AIP is also required for PDE3A-SLFN12 complex formation. Our results provide mechanistic insights into how DNMDP induces PDE3A-SLFN12 complex formation, thereby killing cancer cells with high levels of PDE3A and SLFN12 expression.

Inter-rater agreement of rotator cuff tendon and muscle magnetic resonance imaging parameters evaluated preoperatively and during the first postoperative year following rotator cuff repair.

BACKGROUND: Magnetic resonance imaging (MRI) is standard of care for rotator cuff evaluation, with clinical interpretation usually limited to qualitative judgments. The reliability of MRI-based measurements and scoring systems has been evaluated only preoperatively or ≥6 months following rotator cuff repair, when repairs are in the later stages of healing. This study describes the MRI assessments and inter-rater agreement of various rotator cuff tendon and muscle parameters evaluated preoperatively and 4 times during the first postoperative year. METHODS: Two musculoskeletal radiologists independently assessed MRI scans of 42 patients preoperatively and 3, 12, 26, and 52 weeks after rotator cuff repair. Using standardized reading rules, readers assessed tendon integrity (5-point Sugaya classification), tear dimensions, muscle fat (5-point Goutallier classification) and atrophy (4-point Warner classification), muscle cross-sectional areas, and myotendinous junction distance. Raw exact agreement proportions, κ statistics, and correlation coefficients were used to quantify inter-rater agreement. RESULTS: Readers showed moderate to substantial above-chance agreement in scoring rotator cuff tendon integrity and supraspinatus muscle atrophy and good to excellent agreement on tear dimensions and muscle cross-sectional areas but only fair to moderate agreement for fatty infiltration and myotendinous junction distance. Only fatty infiltration grades evidenced observer bias. Inter-rater agreement did not appear time dependent. CONCLUSION: By use of defined reading rules in a research setting, MRI evaluations of rotator cuff tendon integrity, tear dimensions, muscle atrophy, and cross-sectional areas have reasonable reliability at all time points in the first postoperative year. However, the presence of clinically significant disagreements, even in such favorable circumstances, indicates the need for improved imaging tools for precise rotator cuff evaluation.

Development of a critical-sized ventral hernia model in the pig.

BACKGROUND: The pig is commonly used as a preclinical model for ventral hernia repair. However, no study has verified that an unrepaired surgically induced hernia (control) in the pig does not heal spontaneously but rather develops a persistent hernia. Without such verification in any given model, one cannot draw conclusions on the efficacy of the repair technique investigated. MATERIALS AND METHODS: Three surgically induced hernia models with increasing severity were created in eight pigs. These included 10-cm retrorectus partial-thickness (model 1) and 15-cm preperitoneal full-thickness (model 2) incisional defects and an 8 × 8 cm preperitoneal full-thickness excisional defect (model 3). Postoperative management included use of an abdominal binder, and in some cases, suction drainage, for 2 wk to support the repair and prevent seroma. Models were evaluated for persistence of hernia at 5 wk using clinical and radiographic assessments. RESULTS: All pigs developed clinical hernias after 2 wk of defect creation, but only models 1 and 3 had clinically persistent hernias at 5 wk. At 5 wk, the average defect area was 97 cm2 in model 1, 66 cm2 in model 2, and 245 cm2 in model 3. Dense fibrotic scarring was observed in the models with resolved hernias. CONCLUSIONS: Our results highlight the need to verify an unrepaired hernia injury model does not heal spontaneously prior to using it for hernia repair studies. The partial-thickness incisional model 1 and full-thickness excisional model 3 formed persistent hernias in pigs at 5 wk and should be further explored as models for investigating hernia repair strategies.

Does augmentation with a reinforced fascia patch improve rotator cuff repair outcomes?

BACKGROUND: Scaffold devices are used to augment rotator cuff repairs in humans. While the strength of a novel poly-L-lactic acid-reinforced (human) fascia patch has been documented, it is unclear whether such patches will enhance the strength or likelihood of healing of rotator cuff repairs. QUESTIONS/PURPOSES: In a canine shoulder model, we asked: Do tendon repairs augmented with a reinforced fascia patch have (1) increased biomechanical properties at Time 0 and (2) less tendon retraction and increased cross-sectional area and biomechanical properties after 12 weeks of healing compared to repairs without augmentation? (3) Do the biomechanical properties of tendon repairs reach normal values by 12 weeks of healing? And (4) is the host response associated with use of the reinforced fascia patch biocompatible? METHODS: Eleven dogs underwent bilateral shoulder surgery with partial release and acute repair of the infraspinatus tendon, one shoulder with augmentation and one without augmentation. Repair retraction, cross-sectional area, biomechanical properties, and biocompatibility were assessed at 12 weeks. RESULTS: At Time 0, the mean ± SD ultimate load of augmented repairs was 296 ± 130 N (46% ± 25%) more than nonaugmented repairs, with no difference in stiffness between groups. At 12 weeks, the ultimate load of augmented repairs averaged 192 ± 213 N (15% ± 16%) less than nonaugmented repairs, with no difference in stiffness between groups. At the tendon repair site at 12 weeks, the fascia patch showed a biocompatible host tissue response. CONCLUSIONS: The biomechanical properties of repairs augmented with a reinforced fascia patch demonstrated greater ultimate load at Time 0 than nonaugmented repairs but remained essentially unchanged after 12 weeks of healing, despite improvements in the ultimate load of nonaugmented controls in the same time frame.

Germline Variants in DNA Damage Repair Genes: An Emerging Role in the Era of Precision Medicine in Pancreatic Adenocarcinoma.

Pancreatic adenocarcinoma is a lethal disease that is projected to become the second most common cause of cancer deaths by 2030. The role of adjuvant therapy after surgical resection has been established by several clinical trials to prolong survival and improve outcomes. Multiagent chemotherapy seems to be the most promising approach to counteract early recurrence and improve survival; however, in the era of precision medicine, patient selection and individualized therapy seems to hold the key to desirable superior outcomes. Several cancer susceptibility genes have been proven to be associated with an increased risk of pancreatic cancer, both familial and sporadic cases. The role of genomic profiling for germline variants has been extensive and of limited clinical value, considering their low prevalence in pancreatic ductal adenocarcinoma (PDAC). However, an accumulating body of evidence from several studies in the past decade have successfully shown a recognizable value of germline variants in risk assessment and patient stratification. Recently, anti-PD-1 therapy (pembrolizumab) has been FDA-approved for use in solid malignancies with a Mismatch repair deficiency or high Microsatellite instability. Several trials have evaluated the role of poly (ADP-ribose) polymerase (PARP) inhibitors in patients harboring germline BRCA1/2 mutations. Finally, germline variants in DNA damage response genes and particularly deleterious ones have the potential to guide therapy after surgical resection and serve as biomarkers to predict survival. The dire need to address challenges for applying precision medicine in real-life clinical settings for PDAC patients lies in further characterizing the genetic and molecular processes through translational research.

Agreement between intraoperative and magnetic resonance imaging assessments of rotator cuff pathology and 2 magnetic resonance imaging-based assessments of supraspinatus muscle atrophy.

Background: Magnetic resonance imaging (MRI)-based rotator cuff assessment is often qualitative and subjective; few studies have tried to validate such preoperative assessments. This study investigates relationships of preoperative MRI assessments made by conventional approaches to intraoperative findings of tear type, location, and size or MRI-assessed muscle occupation ratio. Methods: Intraoperatively, surgeons assessed tear type, location, anterior-posterior (AP) width, and medial-lateral length in 102 rotator cuff repair patients. Two musculoskeletal radiologists independently assessed the preoperative MRI scans for these same parameters and supraspinatus muscle atrophy by both Warner classification and quantitative occupation ratio. Exact agreement proportions, kappa statistics, and correlation coefficients were used to quantify agreement relationships. Results: Agreement between MRI readers' and surgeons' observations of tear status averaged 93% with κ = 0.38, and that of tear location averaged 77% with κ = 0.50. Concordance correlations of MRI and intraoperative measures of anterior-posterior and medial-lateral tear length averaged 0.59 and 0.56 across readers, respectively. Despite excellent interrater agreement on Warner classification (exact agreement proportion 0.91) and occupation ratio (concordance correlation 0.93) separately, correlations between these 2 measures were -0.54 and -0.64 for the 2 readers, respectively. Patients with Warner grade 0 had occupation ratios ranging from 0.5 to 1.5. Conclusion: Correlations of preoperative MRI tear dimensions and muscle atrophy assessed by conventional approaches with intraoperatively measured tear dimensions and quantitative occupation ratio, respectively, were only fair. Since tear size and muscle atrophy are known strong predictors of outcomes following rotator cuff repair that may influence treatment decisions, surgeons need to be aware of the limitations of MRI methods. Continued development and validation of quantitative preoperative imaging methods to accurately assess these parameters are needed to improve surgical planning and prognosis.

Consequences of Cryopreservation in Diverse Natural Isolates of Saccharomyces cerevisiae.

Experimental evolution allows the observation of change over time as laboratory populations evolve in response to novel, controlled environments. Microbial evolution experiments take advantage of cryopreservation to archive experimental populations in glycerol media, creating a frozen, living "fossil" record. Prior research with Escherichia coli has shown that cryopreservation conditions can affect cell viability and that allele frequencies across the genome can change in response to a freeze-thaw event. We expand on these observations by characterizing fitness and genomic consequences of multiple freeze-thaw cycles in diploid yeast populations. Our study system is a highly recombinant Saccharomyces cerevisiae population (SGRP-4X) that harbors standing genetic variation that cryopreservation may threaten. We also investigate the four parental isogenic strains crossed to create the SGRP-4X. We measure cell viability over five consecutive freeze-thaw cycles; whereas we find that viability increases over time in the evolved recombinant populations, we observe no such viability improvements in the parental strains. We also collect genome-wide sequence data from experimental populations initially, after one freeze-thaw, and after five freeze-thaw cycles. In the recombinant evolved populations, we find a region of significant allele frequency change on chromosome 15 containing the ALR1 gene. In the parental strains, we find little evidence for new mutations. We conclude that cryopreserving yeast populations with standing genetic variation may have both phenotypic and genomic consequences, though the same cryopreservation practices may have only small impacts on populations with little or no initial variation.

Variability of glenohumeral positioning and bone-to-tendon marker length measurements in repaired rotator cuffs from longitudinal computed tomographic imaging.

BACKGROUND: To address the need for more objective and quantitative measures of tendon healing in research studies, we intend to use computed tomography (CT) with implanted radiopaque markers on the repaired tendon to measure tendon retraction following rotator cuff repair. In our small prior study, retraction at 1-year follow-up averaged 16.1± 5.3 mm and exceeded 10.0 mm in 12 of 13 patients, and thus tendon retraction appears to be a common clinical phenomenon. This study's objectives were to assess, using 5 longitudinal CT scans obtained over 1 year following rotator cuff repair, the variability in glenohumeral positioning because of pragmatic variations in achieving perfect arm repositioning and to estimate the associated measurement variability in bone-to-tendon marker length measurements. METHODS: Forty-eight patients underwent rotator cuff repair with intraoperative placement of radiopaque tendon markers at the repair site. All patients had a CT scan with their arms at the side on the day of surgery and at 3, 12, 26, and 52 weeks postoperatively. Glenohumeral position (defined by the orientation and distance of the humerus with respect to the scapula) and bone-to-tendon marker lengths were measured from each scan. Within-patient variation in glenohumeral position measurements was described by their pooled within-patient standard deviations (SDs), and variation in bone-to-tendon marker lengths by their standard errors of measurement (SEMs) and 95% confidence level minimally detectable distances (MDD95) and changes (MDC95). RESULTS: The mean glenohumeral orientation from the 5 longitudinal CT scans averaged across the 48 patients was 12.6° abduction, 0.4° flexion, and -0.1° internal rotation. Within-patient SDs (95% confidence intervals) of glenohumeral orientation were 3.0° (2.7°-3.4°) in extension/flexion, 5.2° (4.6°-5.8°) in abduction/adduction, and 8.2° (7.3°-9.2°) in internal/external rotation. The SDs of glenohumeral distances were less than 1 mm in any direction. The estimated SEMs of bone-to-tendon lengths were consistent with a common value of 2.4 mm for any of the tendon markers placed across the repair, with MDD95 of 4.7 mm and MDC95 of 6.7 mm. CONCLUSION: Apparent tendon retraction of 5 mm or more, when measured as the distance from a tendon marker's day of surgery location to its new location on a volumetrically registered longitudinal CT scan, may be considered above the usual range of measurement variation. Tendon retraction measured using implanted radiopaque tendon markers offers an objective and sufficiently reliable means for quantifying the commonly expected changes in structural healing following rotator cuff repair.

Biodegradable hyaluronan hydrogel coatings on acellular dermis grafts-A potential strategy to improve biologic graft durability in hernia repair application.

Biologic grafts used in hernia repair undergo rapid cellular infiltration and remodeling, but their premature degradation often results in hernia recurrence. We hypothesize that a temporary barrier that prevents infiltration of acute inflammatory cells into the graft during the initial 4 weeks of implantation could mitigate graft degradation. The purpose of this study is to design tyramine-substituted hyaluronan (THA) hydrogel coatings with tunable degradation properties, as a means to develop a resorbable barrier for human acellular dermis grafts (HADM). THA plugs prepared at different cross-linking densities, by varying cross-linking agent concentration (0.0001-0.0075% H2 O2 ), demonstrated varying rates of in vitro degradation (25 U/mL hyaluronidase, 48 h). Based on these results, HADM grafts were coated with THA at three cross-linking densities (0.0001%, 0.00075%, and 0.003% H2 O2 ) and THA coating degradation was evaluated in vitro (25 U/mL hyaluronidase, 48 h) and in vivo (rat intraperitoneal implantation, 1-4 weeks). THA coatings degraded in vitro and in vivo with the lowest cross-linking density (0.0001% H2 O2 ), generally showing greater degradation as evidenced by significant decrease in coating cross-sectional area. However, all three coatings remained partially degraded after 4 weeks of in vivo implantation. Alternate strategies to accelerate in vivo degradation of THA coatings are required to allow investigation of the study hypothesis. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 107B:2664-2672, 2019.

A novel radiopaque tissue marker for soft tissue localization and in vivo length and area measurements.

PURPOSE: The purpose of the study was to describe the characteristics and demonstrate proof-of-concept and clinical use of a barium sulfate infused polypropylene radiopaque tissue marker for soft tissue localization and in vivo measurement of lengths and areas. METHODS: Marker mechanical properties were evaluated by tensile tests. Biocompatibility was evaluated following 8-12 weeks' implantation in a pig model. Proof-of-concept of marker application was performed in a human cadaveric shoulder model, and methods for CT imaging and measurement of dimensions were established. Lastly, the method of clinical use of the markers was described in one patient undergoing arthroscopic rotator cuff repair (RCR). RESULTS: The radiopaque markers had a tensile strength of 28 ±4.7 N and were associated with minimal to mild inflammatory tissue reaction similar to polypropylene control. CT-based measurements showed relatively high precisions for lengths (0.66 mm), areas (6.97 mm2), and humeral orientation angles (2.1°) in the cadaveric model, and demonstrated 19 ±3 mm medio-lateral tendon retraction and 227 ±3 mm2 increase in tendon area in the patient during 26 weeks following RCR. No radiographic leaching, calcification or local adverse events were observed. CONCLUSIONS: The radiopaque tissue marker was biocompatible and had adequate strength for handling and affixation to soft tissues using standard suturing techniques. The marker could be used with low-dose, sequential CT imaging to quantitatively measure rotator cuff tendon retractions with clinically acceptable accuracy. We envision the radiopaque tissue marker to be useful for soft tissue localization and in vivo measurement of tissue and organ dimensions following surgery.

Regional variability, processing methods, and biophysical properties of human fascia lata extracellular matrix.

This study aims to assess the regional variability, processing methods, mechanical, biochemical, and cellular properties of human fascia lata as a scaffold for soft tissue repair and tissue engineering applications. Ten pairs of fascia lata (donor age 18-55) were used. One fascia patch from each pair was used to assess the geometric and biomechanical variability of fresh fascia. The other from each pair was subjected to 1 of 2 allograft processing methods: antibiotic soak alone or acellularization plus antibiotic soak. Stiffness, modulus, hydroxyproline, chondroitin/dermatan sulfate glycosaminoglycan (CSDS GAG), and DNA content were quantified in fascia from fresh and treated groups. The effect of location was not significant for thickness or stiffness within a 6 x 12 cm2 region of the iliotibial tract of fresh human fascia lata. Processing did not significantly change the stiffness, modulus, or CSDS GAG content of fascia ECM. However, hydroxyproline (collagen) content is significantly reduced in acellularized fascia, probably reflecting a removal of soluble collagen during the treatment (p < 0.02). Processing reduced the DNA content of fresh fascia approximately 10-fold (p < 0.001). The mechanical, chemical and ultrastructural similarities between fascia lata and tendon may make fresh or processed fascia an attractive ECM scaffold for soft tissue, particularly tendon, repair.

Assessment of the canine model of rotator cuff injury and repair.

Animal shoulder models are used to systematically investigate the factors influencing rotator cuff injury and repair. Each model has advantages and disadvantages that must be considered in the context of the specific research questions being asked. This study evaluated the utility of the canine model for studies of acute, full-thickness rotator cuff tendon injury and repair. We found that time-zero failure load is dependent on the suture type and configuration used for repair. Acute, full-width tendon repairs fail anatomically within the first days after surgery in the canine model, regardless of suture type, suture configuration, or postoperative protocol. Robust scar tissue forms in the gap between the failed tendon end and the humerus, which can be visually, mechanically, and histologically misconstrued as tendon if an objective test of repair connectivity is not performed. We conclude that a full-width injury and repair model in the canine will provide a rigorous test of whether a new repair strategy or postoperative protocol, such as casting or temporary muscle paralysis, can maintain repair integrity in a high-load environment. Alternatively, a partial-width tendon injury model allows loads to be shared between the tendon repair and the remaining intact portion of the infraspinatus tendon and prohibits complete tendon retraction. Thus a partial-width injury in the canine may model the mechanical environment of many single tendon tears in the human injury condition and warrants further investigation.

Exploratory study on the effect of osteoactivin on muscle regeneration in a rat volumetric muscle loss model.

Wounds causing extensive injury loss of muscle, also known as volumetric muscle loss (VML), are frequently associated with high-energy civilian trauma and combat-related extremity injuries. Currently, no effective clinical therapy is available for promoting de novo muscle tissue regeneration to restore muscle function following VML. Recent studies have shown evidence that osteoactivin (OA), a transmembrane glycoprotein, has the ability to prevent skeletal muscle atrophy in response to denervation. Therefore the objective of this study is to investigate the potential regenerative effect of OA embedded and delivered via a cross-linked gelatin hydrogel within a volumetric tibialis anterior muscle defect in a rat model. After 4 weeks, however, no evidence for muscle formation was found in defects treated with either low (5 µg/ml) or high (50 µg/ml) OA. It is possible that a different delivery scaffold, delivery kinetics, or OA concentration may have yielded an alternate outcome, or it is also possible that the spaciostructural environment of VML, or the local (versus systemic) delivery of OA, simply does not support any potential regenerative activity of OA in VML. Together with prior work, this study demonstrates that an efficacious and scalable therapy for regenerating muscle volume and function in VML remains a veritable clinical challenge worthy of continued future research efforts.

Assessment of Human Acellular Dermis Graft in Porcine Models for Ventral Hernia Repair.

OBJECTIVE: Preclinical evaluation of hernia meshes is commonly performed in porcine models. We recently developed two surgically induced porcine hernia models-an incisional and an excisional model-that formed persistent hernias in the absence of graft repair. Herein, we investigate if these models will continue to form a hernia after graft repair. MATERIALS AND METHODS: Ten pigs were used to create two hernia models-a 10-cm partial-thickness incisional defect (n = 5) and an 8 × 8-cm full-thickness excisional defect (n = 5). The defects were repaired using a 12 × 12-cm human acellular dermis graft placed in a preperitoneal/retrorectus sublay position and fixed using transfascial sutures. Postoperative management included the use of suction drainage for 1 week and an abdominal binder for 4 weeks in the more severe excisional model. Hernia development was assessed clinically, and hernia defect size and volume were measured using postoperative computed tomography (CT) imaging over 12 weeks. Radiographic inflation testing (2 L inflation), biaxial mechanical testing, and histological evaluation were also performed at 12 weeks. RESULTS: All pigs with the excisional model, but none with the incisional model, developed a clinically relevant hernia. At the end of 12 weeks, the excisional model had a significantly greater hernia defect size (259 ± 51 cm2 vs. 47 ± 16 cm2) and repair volume (865 ± 414 cm3 vs. 85 ± 52 cm3) compared with the incisional model. The excisional model also showed an order of magnitude greater increase in repair volume (280 cm3 vs. 47 cm3) compared with the incisional model upon 2 L inflation. Furthermore, the excisional model showed a trend of having higher dilatational strain at average biaxial load of 250 N and lower stiffness compared with the incisional model. The excisional model had a thin, hypercellular hernia sac spanning the defect, whereas the incisional model had a thick densely fibrotic scar bridging the defect. CONCLUSION: The 8 × 8-cm excisional defect model, together with appropriate postoperative wound management, in the pig model is recommended for preclinical investigation of different grafts for hernia repair. Novel CT imaging and biomechanical testing methods are recommended to measure functional outcomes of hernia repair in preclinical models.

Commercial extracellular matrix scaffolds for rotator cuff tendon repair. Biomechanical, biochemical, and cellular properties.

BACKGROUND: We are not aware of any in vitro study comparing the biomechanical, biochemical, and cellular properties of commercial extracellular matrix materials marketed for rotator cuff tendon repair. In this study, the properties of GraftJacket, TissueMend, Restore, and CuffPatch were quantified and compared with each other. The elastic moduli were also compared with that of normal canine infraspinatus tendon. METHODS: Samples were tested from different manufacturing lots of four materials: GraftJacket (ten lots), TissueMend (six), Restore (ten), and CuffPatch (six). The Kruskal-Wallis test was used to compare thickness, stiffness, and modulus as well as hydroxyproline, chondroitin/dermatan sulfate glycosaminoglycan, hyaluronan, and DNA contents among these matrices. The moduli of the extracellular matrices were also compared with those of normal canine infraspinatus tendon. RESULTS: All four extracellular matrices required 10% to 30% stretch before they began to carry substantial load. Their maximum moduli were realized in their linear region at 30% to 80% strain. The elastic moduli of all four commercial matrices were an order of magnitude lower than that of canine infraspinatus tendon. TissueMend had significantly higher DNA content than the other three matrices (p<0.0001), although both Restore and GraftJacket also had measurable amounts of DNA. CONCLUSIONS: Our data demonstrate chemical and mechanical differences among the four commercial extracellular matrices that we evaluated. Probably, the source (dermis or small intestine submucosa), species (human, porcine, or bovine), age of the donor (fetal or adult), and processing of these matrices all contribute to the unique biophysical properties of the delivered product. The biochemical composition of commercial extracellular matrices is similar to that of tendon. However, the elastic moduli of these materials are an order of magnitude lower than that of tendon, suggesting a limited mechanical role in augmentation of tendon repair.

Development of an Arthroscopic Joint Capsule Injury Model in the Canine Shoulder.

BACKGROUND: The natural history of rotator cuff tears can be unfavorable as patients develop fatty infiltration and muscle atrophy that is often associated with a loss of muscle strength and shoulder function. To facilitate study of possible biologic mechanisms involved in early degenerative changes to rotator cuff muscle and tendon tissues, the objective of this study was to develop a joint capsule injury model in the canine shoulder using arthroscopy. METHODS: Arthroscopic surgical methods for performing a posterior joint capsulectomy in the canine shoulder were first defined in cadavers. Subsequently, one canine subject underwent bilateral shoulder joint capsulectomy using arthroscopy, arthroscopic surveillance at 2, 4 and 8 weeks, and gross and histologic examination of the joint at 10 weeks. RESULTS: The canine subject was weight-bearing within eight hours after index and follow-up surgeries and had no significant soft tissue swelling of the shoulder girdle or gross lameness. Chronic synovitis and macroscopic and microscopic evidence of pathologic changes to the rotator cuff bony insertions, tendons, myotendinous junctions and muscles were observed. CONCLUSIONS: This study demonstrates feasibility and proof-of-concept for a joint capsule injury model in the canine shoulder. Future work is needed to define the observed pathologic changes and their role in the progression of rotator cuff disease. Ultimately, better understanding of the biologic mechanisms of early progression of rotator cuff disease may lead to clinical interventions to halt or slow this process and avoid the more advanced and often irreversible conditions of large tendon tears with muscle fatty atrophy.

Investigating muscle regeneration with a dermis/small intestinal submucosa scaffold in a rat full-thickness abdominal wall defect model.

Repair of large complex ventral wall hernias is challenging and outcomes are often poor due to hernia recurrence and compromised abdominal wall function. Currently, biological grafts are used to assist in repairing these complex hernias. Dermis grafts are often chosen because of their mechanical characteristics; however, dermis does not have the ability to promote the muscle regeneration needed to regain abdominal wall function. In contrast, small intestinal submucosa (SIS) grafts have been shown to promote muscle generation in volumetric muscle loss (VML) models. Hence, the objective of this study is to investigate the extent to which SIS grafts can be used together with dermis grafts to repair and promote muscle regeneration in a full-thickness abdominal wall defect in a rat model. The dermis layer is intended to mechanically bridge the defect and support constructive tissue remodeling while the SIS is intended to degrade and promote neo-muscle formation. After 16 weeks of implantation, we found only a small amount of vascularized muscle (<10% of the defect area) in the repaired defects. No significant difference in defect muscle area was found between the groups receiving the dermis + SIS scaffolds and the control (dermis alone) group. Our findings indicate that the SIS constructions investigated could not promote appreciable muscle regeneration in this rigorous animal model of VML and incomplete abdominal closure. Future investigation into combination scaffold, cell and molecular therapies would be warranted to address the need for functional muscle regeneration in challenging clinical conditions such as complex abdominal wall repair.

Human fascia lata ECM scaffold augmented with immobilized hyaluronan: inflammatory response and remodeling in the canine body wall and shoulder implantation sites.

We postulate that immobilization of tyramine-substituted hyaluronan (THA) into an extracellular matrix (ECM) scaffold may be a strategy to promote an anti-inflammatory response to the ECM. Further, we posit that the implantation site could influence the inflammatory response and remodeling of an ECM scaffold. Eight beagles underwent implantation of fascia ECM grafts, treated with either immobilized low molecular weight (57 kDa) THA or water only, in both the shoulder injury and body wall sites. Dogs were euthanized at 12 weeks and fascia grafts harvested en bloc for histology. Grafts implanted at the body wall had significantly higher inflammatory cell infiltrate and vascularity, and significantly lower retardance (collagen density), than grafts at the shoulder, suggestive of a more intense, persistent, and perhaps degradative inflammatory and remodeling response at the body wall than shoulder injury site in the canine model. However, the presence of immobilized low MW THA had no effect on the inflammation response or remodeling of fascia ECM compared to water-treated controls. Importantly, these results suggest that the inflammatory response and remodeling of biomaterial implants depends on the location of implantation and therefore our animal models need to be carefully chosen. Further, the potential anti-inflammatory advantages of hyaluronan (HA) in wound healing do not appear to be realized when presenting it to the host as non-degradable hydrogel even if its capacity for binding HA binding protein is maintained. Further study treating ECM with uncross-linked (free) HA or immobilized low MW THA as a means to deliver free HA or other biomolecules to a surgical repair site is warranted.

Host response to xenograft ECM implantation is not different between the shoulder and body wall sites in the rat model.

In the context of tendon and ligament repair, mechanical loading and the presence of joint synovial fluid are known to profoundly influence the form and function of the repair tissue and potentially the host response to biomaterials. Previously, we demonstrated that a xenograft extra cellular matrix (ECM) scaffold implanted in the rat shoulder elicited a unique host response from that seen in the body wall. However, the host response to xenografts implanted in shoulders with a tendon/capsule injury was not different from xenografts implanted in shoulders with no injury. In the current study, we hypothesized that varying clinically relevant surgical and environmental factors would introduce significant differences in host response to xenograft implantation at the shoulder. Contrary to our hypothesis, we found no significant differences in host response between any shoulder implantation conditions or between shoulder and body wall implantation in the rat model. These findings suggest that there is no advantage to using an orthotopic shoulder model to investigate the host response to rotator cuff scaffold materials in the rat model, and due to the insensitivity of its host response to various clinically relevant surgical conditions, may suggest that the rat does not provide a surrogate for directly translating the host response to biomaterials to the human application.