Genetically engineered animal models for Marfan syndrome: challenges associated with the generation of pig models for diseases caused by haploinsufficiency.

Recent developments in reproductive biology have enabled the generation of genetically engineered pigs as models for inherited human diseases. Although a variety of such models for monogenic diseases are currently available, reproduction of human diseases caused by haploinsufficiency remains a major challenge. The present study compares the phenotypes of mouse and pig models of Marfan syndrome (MFS), with a special focus on the expressivity and penetrance of associated symptoms. Furthermore, investigation of the gene regulation mechanisms associated with haploinsufficiency will be of immense utility in developing faithful MFS pig models.

Stride-Phase Kinematic Parameters That Predict Peak Elbow Varus Torque.

BACKGROUND: During baseball pitching, a high amount of elbow varus torque in the arm cocking-to-acceleration phase is thought to be a biomechanical risk factor for medial elbow pain and injury. The biomechanics of the stride phase may provide preparation for the arm cocking-to-acceleration phase that follows it. PURPOSE: To determine the kinematic parameters that predict peak elbow varus torque during the stride phase of pitching. STUDY DESIGN: Descriptive laboratory study. METHODS: Participants were 107 high school baseball pitchers (age range, 15-18 years) without shoulder or elbow problems. Whole-body kinematics and kinetics during fastball pitching were analyzed using 3-dimensional measurements from 36 retroreflective markers. A total of 26 kinematic parameters of the upper and lower limbs during the stride phase leading up to the stride foot contact were extracted for multiple regression analysis to assess their combined effect on the magnitude of peak elbow varus torque. RESULTS: Increased wrist extension, elbow pronation, knee flexion on the leading leg, knee extension on the trailing leg at stride foot contact, and upward displacement of the body's center of mass in the stride phase were significantly correlated with decreased peak elbow varus torque (all P < .05). Moreover, 38% of the variance in peak elbow varus torque was explained by a combination of these 5 significant kinematic variables (P < .001). CONCLUSION: We found that 5 kinematic parameters during the stride phase and the combination of these parameters were associated with peak elbow varus torque. The stride phase provides biomechanical preparation for pitching and plays a key role in peak elbow varus torque in subsequent pitching phases. CLINICAL RELEVANCE: The present data can be used to screen pitching mechanics with motion capture assessment to reduce peak elbow varus torque. Decreased peak elbow varus torque is expected to reduce the risk of elbow medial pain and injury.

Influence of advanced glycation end products on rotator cuff.

BACKGROUND: Most rotator cuff tears are the result of age-related degenerative changes, but the mechanisms underlying these changes have not been reported. Recently, advanced glycation end products (AGEs) have been regarded as an important factor in senescence. Therefore, we hypothesized that AGEs would have detrimental effects on rotator cuff-derived cells. In this study, we investigated the influence of AGEs on rotator cuff-derived cells in vitro and ex vivo. METHODS: Rotator cuff-derived cells were obtained from human supraspinatus tendons. The cells were cultured in the following media: (1) regular medium with 500 µg/mL AGEs (High-AGEs), (2) regular medium with 100 µg/mL AGEs (Low-AGEs), and (3) regular medium alone (Control). Cell viability, secretion of vascular endothelial growth factor, and the expressions of hypoxia-inducible factor-1α, reactive oxygen species, and apoptosis were assessed after cultivation. An ex vivo tissue culture with AGEs was also performed to measure the tensile strength. RESULTS: Cell viability in the High-AGEs group was significantly suppressed relative to that in the Controls. The amount of vascular endothelial growth factor secretion was significantly greater in the High- and Low-AGEs groups than in the Controls. Immunofluorescence stain demonstrated enhancement of hypoxia-inducible factor-1α and reactive oxygen species expressions and cell apoptosis in the High- and Low-AGEs groups relative to that in the Controls. In ex vivo mechanical testing, tensile strength was significantly higher in the Control group than in the AGEs groups. DISCUSSION: These results indicated that AGEs caused age-related degenerative rotator cuff changes. The reduction of AGEs might prevent rotator cuff senescence-related degeneration.

Estimation of Shoulder Behavior From the Viewpoint of Minimized Shoulder Joint Load Among Adolescent Baseball Pitchers.

BACKGROUND: During pitching, an overloaded joint reaction force exerted on the shoulder and excessive shoulder horizontal abduction at ball release are considered risk factors causing anterior shoulder pain for young baseball pitchers. Hypothesis/Purpose: The first aim was to examine the relationship between shoulder horizontal abduction position and force on the shoulder at ball release. The second was to identify the relative rotational position of the shoulder and the range of shoulder motion at ball release that minimize force on the shoulder. It was hypothesized that the amount of force on the shoulder would be exacerbated by excessive shoulder horizontal abduction. STUDY DESIGN: Descriptive laboratory study. METHODS: Participants were 183 adolescent baseball pitchers (mean ± SD age, 15.5 ± 1.2 years) without shoulder/elbow problems. Each pitcher threw 5 fastballs to a catcher behind a home plate. The kinematics and kinetics of the throwing shoulder during fastball pitching were calculated with 3-dimensional measurements from 36 reflective markers. In data analysis, the correlations were calculated between the relative rotational positions of the shoulder (abduction, horizontal adduction-abduction) and the forces on the shoulder (anterior-posterior, proximal, and superior-inferior) at ball release. Subsequently, the specific rotational position and range of motion of the shoulder at ball release that minimized forces on the shoulder were determined. RESULTS: Statistically significant correlations were identified between the magnitude of superior-inferior force on the shoulder and shoulder abduction position ( R2 = 0.44, P < .001) as well as between the magnitude of anterior-posterior force on the shoulder and shoulder horizontal adduction-abduction position ( R2 = 0.72, P < .001). Minimal anterior-posterior and superior-inferior forces were obtained with a combination of 80.6° of shoulder abduction and 10.7° of shoulder horizontal adduction. Any deviation >5° from this position significantly increased the anterior-posterior and superior-inferior forces on the shoulder. CONCLUSION: Increasing shoulder horizontal abduction position significantly increased the magnitude of anterior force on the shoulder at ball release. The combination of 80.6° of shoulder abduction and 10.7° of horizontal shoulder adduction minimized the shear forces on the shoulder at this point. CLINICAL RELEVANCE: The present data can be useful for screening pitching technique to prevent shoulder pain and injury with motion capture assessment.

Rotator cuff tear healing process with graft augmentation of fascia lata in a rabbit model.

BACKGROUND: Fascia lata augmentation of massive rotator cuff tears has shown good clinical results. However, its biological effect during the early healing process is not clearly understood. The purpose of the study was to evaluate the biological efficacy of fascia lata augmentation during the early healing process of rotator cuff tears using a rabbit rotator cuff defect model. METHODS: The infraspinatus tendon was resected from the greater tuberosity of a rabbit to create a rotator cuff tear. The tendon edge was directly sutured to the humeral head. The rotator cuff repaired site was augmented with a fascia lata autograft (augmentation group, group A). The rotator cuff defect in the contralateral shoulder was repaired without augmentation (reattachment group, group R). A group with intact rotator cuff was set as the control group. Histological examinations and mechanical analysis were conducted 4 and 8 weeks postoperatively. RESULTS: In the HE staining, the tendon maturing score of group A was higher than that of group R at 4 weeks postoperatively. In the safranin O staining, proteoglycan staining at the repaired enthesis in group A at 4 weeks postoperatively was stronger than that in group R. Picrosirius red staining showed that type III and type I collagen in group A was more strongly expressed than that in group R at 4 weeks postoperatively. The ultimate failure load of the infraspinatus tendon-humeral head complex in group A was statistically higher than that in group R at 4 weeks postoperatively. The ultimate failure load of group A was similar to that of the control group. CONCLUSION: The biological and mechanical contribution of fascia lata augmentation for massive rotator cuff tears was analyzed in this study. Type III collagen was reported to be expressed during the tendon healing process. Although the biological action similar to natural ligament healing occurred around the fascia lata grafts, type III collagen was gradually replaced by type I collagen as the tissue matured. Our results suggest that fascia lata augmentation could stimulate biological healing and provide initial fixation strength of the repaired rotator cuff.

Characteristics and Clinical Outcomes in Overhead Sports Athletes after Rotator Cuff Repair.

Rotator cuff tears in young overhead sports athletes are rare. The pathomechanism causing rotator cuff tears in young overhead athletes is different from that in aged patients. The purpose of this study was to investigate rotator cuff tear characteristics in young overhead sports athletes to reveal the pathomechanism causing these injuries. This study included 25 overhead sports athletes less than 30 years old with atraumatic rotator cuff tears necessitating repair. Rotator cuff tear characteristics were evaluated intraoperatively, including rotator cuff tear shape and injured rotator cuff tendon. Clinical outcome measures were assessed before surgery and at the final follow-up. In this study, 22 patients reported minimal to no shoulder pain and returned to sports without significant complaints at last follow-up. The isolated infraspinatus tendon was most often injured; the incidence rate of the tear at this site was 32% (8 cases). In the deceleration phase of overhead motion, the eccentric contraction force of the ISP (infraspinatus) tendon peaks and the increased load leads to injury at the ISP tendon. The pathomechanism of rotator cuff injuries in young overhead athletes might be not only internal or subacromial impingement, but also these mechanisms.

Effect of platelet-rich plasma on degeneration change of rotator cuff muscles: In vitro and in vivo evaluations.

Atrophy with fatty degeneration is often seen in rotator cuff muscles with torn tendons. PRP has been reported to enhance tissue repair processes after tendon ruptures. However, the effect of PRP on atrophy and fatty degeneration of the muscle is not yet known. The aim of this study is to examine the effect of PRP on degeneration change of rotator cuff muscles in vitro and in vivo. A murine myogenic cell line and a rat rotator cuff tear model were used in this study and PRP was administrated into subacromial space which is widely used in clinical practice. In in vitro study, administration of PRP to C2C12 cells stimulated cell proliferation while inhibited both myogenic and adipogenic differentiation. In in vivo study, administration of PRP suppressed Oil Red-O positive lipid droplet formation. The expression of adipogenic genes was also decreased by PRP administration. In conclusion, PRP promoted proliferation of myoblast cells, while inhibiting adipogenic differentiation of myoblast cells and suppressing fatty degeneration change in rat torn rotator cuff muscles. Further investigations are needed to determine the clinical applicability of the PRP. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:1806-1815, 2017.

Rotator cuff repair using cell sheets derived from human rotator cuff in a rat model.

To achieve biological regeneration of tendon-bone junctions, cell sheets of human rotator-cuff derived cells were used in a rat rotator cuff injury model. Human rotator-cuff derived cells were isolated, and cell sheets were made using temperature-responsive culture plates. Infraspinatus tendons in immunodeficient rats were resected bilaterally at the enthesis. In right shoulders, infraspinatus tendons were repaired by the transosseous method and covered with the cell sheet (sheet group), whereas the left infraspinatus tendons were repaired in the same way without the cell sheet (control group). Histological examinations (safranin-O and fast green staining, isolectin B4, type II collagen, and human-specific CD31) and mRNA expression (vascular endothelial growth factor; VEGF, type II collagen; Col2, and tenomodulin; TeM) were analyzed 4 weeks after surgery. Biomechanical tests were performed at 8 weeks. In the sheet group, proteoglycan at the enthesis with more type II collagen and isolectin B4 positive cells were seen compared with in the control group. Human specific CD31-positive cells were detected only in the sheet group. VEGF and Col2 gene expressions were higher and TeM gene expression was lower in the sheet group than in the control group. In mechanical testing, the sheet group showed a significantly higher ultimate failure load than the control group at 8 weeks. Our results indicated that the rotator-cuff derived cell sheet could promote cartilage regeneration and angiogenesis at the enthesis, with superior mechanical strength compared with the control. Treatment for rotator cuff injury using cell sheets could be a promising strategy for enthesis of tendon tissue engineering. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:289-296, 2017.

Can Platelet-Rich Plasma Protect Rat Achilles Tendons From the Deleterious Effects of Triamcinolone Acetonide?

BACKGROUND: Triamcinolone acetonide (TA) injections are widely used for tendinitis but have deleterious effects, including tendon degeneration or tendon rupture. PURPOSE: To investigate whether adding platelet-rich plasma (PRP), a blood fraction that participates in tissue repair processes, to TA can prevent its deleterious effects. STUDY DESIGN: Controlled laboratory study. METHODS: Rat Achilles tendons were injected with TA, TA + PRP, PRP alone, or saline (control). Biomechanical testing and histological analyses were performed on Achilles tendons 1 week after injections. RESULTS: The maximum failure loads in the control, TA, TA + PRP, and PRP groups were 31.7 ± 2.3, 19.0 ± 3.6, 31.0 ± 7.1, and 30.2 ± 6.8 N, respectively. The tendon stiffness in the control, TA, TA + PRP, and PRP groups was 12.1 ± 1.8, 7.5 ± 1.8, 11.0 ± 2.8, and 11.3 ± 2.5 N/mm, respectively. The maximum failure load and stiffness were significantly lower in the TA group compared with the other 3 groups. There was no significant difference between the TA + PRP and control groups. Cell invasions, vacuolation, collagen attenuation, and increased type III collagen expression were histologically observed in the TA group; however, these changes were prevented by the simultaneous administration of PRP. CONCLUSION: Administering PRP may prevent deleterious effects caused by TA; therefore, PRP may be used as a protective agent in clinical situations. CLINICAL RELEVANCE: PRP can be useful as a protective agent for sports injury patients receiving local corticosteroid injections.

Rotator Interval Lesion and Damaged Subscapularis Tendon Repair in a High School Baseball Player.

In 2013, a 16-year-old baseball pitcher visited Nobuhara Hospital complaining of shoulder pain and limited range of motion in his throwing shoulder. High signal intensity in the rotator interval (RI) area (ball sign), injured subscapularis tendon, and damage to both the superior and middle glenohumeral ligaments were identified using magnetic resonance imaging (MRI). Repair of the RI lesion and partially damaged subscapularis tendon was performed in this pitcher. During surgery, an opened RI and dropping of the subscapularis tendon were observed. The RI was closed in a 90° externally rotated and abducted position. To reconfirm the exact repaired state of the patient, arthroscopic examination was performed from behind. However, suture points were not visible in the >30° externally rotated position, which indicates that the RI could not be correctly repaired with the arthroscopic procedure. One year after surgery, the patient obtained full function of the shoulder and returned to play at a national convention. Surgical repair of the RI lesion should be performed in exactly the correct position of the upper extremity.

Application of Pain Quantitative Analysis Device for Assessment of Postoperative Pain after Arthroscopic Rotator Cuff Repair.

PURPOSE: The PainVision™ system was recently developed for quantitative pain assessment. Here, we used this system to evaluate the effect of plexus brachialis block on postoperative pain after arthroscopic rotator cuff repair. METHODS: Fifty-five patients who underwent arthroscopic rotator cuff repair were included in this study. First 26 cases received no plexus brachialis block (control group), and the next 29 cases received the plexus brachialis block before surgery (block group). Patients completed the visual analog scale at 4, 8, 16, and 24 hours after surgery, and the intensity of postoperative pain was assessed with PainVision™ at 16 hours. The postoperative use of non-steroidal anti-inflammatory agents was also recorded. RESULTS: The pain intensity at 16 hours after surgery assessed by PainVision™ was significantly lower in the block group than in the control group (block, 252.0 ± 47.8, control, 489.0 ± 89.1, P < 0.05). However, there were no differences in the VAS values at 16 hours between the 2 groups (block, 4.3 ± 0.6, control, 5.7 ± 0.4, P = N.S.). The pain intensity and VAS at 16 hours after surgery were highly correlated (r = 0.59, P = 0.006 in the block group and r = 0.62, P = 0.003 in the control group). The effect size of the assessment by PainVision™ was bigger than that of VAS (r=0.31 in VAS and 0.51 in Pain vision). CONCLUSION: The PainVision™ system could be useful to evaluate postoperative pain because it enables the quantification and comparison of pain intensity independent of individual pain thresholds.

Concurrent rotator cuff tear and axillary nerve palsy associated with anterior dislocation of the shoulder and large glenoid rim fracture: a "terrible tetrad".

We present a case of concurrent rotator cuff tear and axillary nerve palsy resulting from anterior dislocation of the shoulder and a large glenoid rim fracture-a "terrible tetrad." A 61-year-old woman fell on her right shoulder. Radiographs showed anterior dislocation of the shoulder with a glenoid rim fracture, and an MRI two months after injury revealed a rotator cuff tear. Upon referral to our hospital, physical and electrophysiological examinations revealed axillary nerve palsy. The axillary nerve palsy was incomplete and recovering, and displacement of the glenoid rim fracture was minimal and already united; therefore, we surgically repaired only the rotator cuff tear three months after injury. The patient recovered satisfactorily following the operation. In patients whose axillary nerve palsy is recovering, surgeons should consider operating on rotator cuff tears in an attempt to prevent rotator cuff degeneration.

Temporary inductions of matrix metalloprotease-3 (MMP-3) expression and cell apoptosis are associated with tendon degeneration or rupture after corticosteroid injection.

Corticosteroid injections are widely used to treat enthesopathy and tendinitis, but are also associated with possible side effects, such as tendon degeneration or rupture. However, the mechanism of tendon degeneration or rupture after corticosteroid injection remains controversial. The purpose of this study was to reveal the mechanism of tendon degeneration or rupture after injection of triamcinolone acetonide (TA) or prednisolone (PSL). Forty-two rats were divided into 3 groups: A normal saline injection group (control group), a TA injection group, and a PSL injection group; the normal saline or corticosteroid was injected around the Achilles tendon. One or 3 weeks after injection, the tendons were subjected to biomechanical testing and histological analysis. At 1 week, the biomechanical strength was significantly lower in the corticosteroid groups. Histological analysis, at 1-week post-injection, showed collagen attenuation, increased expression of MMP-3 and apoptotic cells in the corticosteroid groups. The histological changes and biomechanical weaknesses of the tendon were not seen at 3 weeks. These alterations appeared to be involved in tendon degeneration or rupture after corticosteroid injection.

Platelet-rich plasma protects rotator cuff-derived cells from the deleterious effects of triamcinolone acetonide.

Triamcinolone acetonide (TA) injections are widely used to treat enthesopathy, but they may induce adverse effects such as tendon impairment and rupture. Platelet-rich plasma (PRP) is a blood fraction containing high platelet concentrations and various growth factors that play a role in tissue repair processes. The purpose of this study is to investigate whether TA has deleterious effects on human rotator cuff-derived cells, and if PRP can protect these cells from the effects of TA. Human rotator cuff-derived cells were cultured with and without TA and PRP, and the culture without any additive served as the control. Cell morphology was assessed at days 7 and 21. Cell viability was evaluated at days 1, 7, 14, and 21 by a water-soluble tetrazolium salt assay. Induction of apoptosis was measured by immunofluorescence staining and flow cytometry at day 7. Induction of cleaved caspase-3 was measured by immunofluorescence staining at day 7. The cells cultured with TA had a flattened and polygonal shape at day 7. The cells cultured with both TA and PRP were similar in appearance to control cells. Exposure to TA also significantly decreased cell viability, but cell viability did not decrease when PRP was added along with TA. The number of apoptotic cells increased with TA exposure, while addition of PRP prevented cell apoptosis. In conclusion, the deleterious effect of TA was prevented by PRP, which can be used as a protective agent for patients receiving local TA injections.

Long-term outcome of trabeculotomy for the treatment of developmental glaucoma.

OBJECTIVE: To elucidate long-term outcome of trabeculotomy in primary and secondary developmental glaucoma. METHODS: One hundred forty-nine eyes of 89 patients with developmental glaucoma who underwent trabeculotomy were retrospectively studied. Intraocular pressure (IOP), success probabilities, visual acuities, and visual field were determined during follow-up and at the final visit. RESULTS: The mean +/- SD IOP of 112 eyes with primary developmental glaucoma at the final visit with an mean +/- SD follow-up period of 9.5 +/- 7.1 years was 15.6 +/- 5.0 mm Hg. The average IOP for 37 eyes with secondary developmental glaucoma was 16.7 +/- 4.2 mm Hg. One hundred eyes (89.3%) with primary developmental glaucoma were defined as achieving success at the final visit. Complete and qualified successes were achieved in 71 eyes (63.4%) and 29 eyes (25.9%), respectively. Visual acuities were 20/40 or better in 78 (59.5%) of 131 eyes examined and were poorer than 20/200 in 32 eyes (24.4%). The causes of poor visual acuities were mainly progression of glaucoma, including delay of detection of onset or surgery and amblyopia. Eyes with glaucoma that existed before 2 months of age or eyes that needed several trabeculotomies were considered to have poor visual acuity. Visual fields were classified as normal or almost normal in 21 (44.7%) of 47 eyes. CONCLUSIONS: Trabeculotomy for developmental glaucoma is effective over a long time. There is a fairly good prognosis for visual function of eyes with developmental glaucoma with early detection of the onset, proper treatment, and proper management after trabeculotomy.