A Novel Tendon Injury Model, Induced by Collagenase Administration Combined with a Thermo-Responsive Hydrogel in Rats, Reproduces the Pathogenesis of Human Degenerative Tendinopathy.

Patellar tendinopathy is a common clinical problem, but its underlying pathophysiology remains poorly understood, primarily due to the absence of a representative experimental model. The most widely used method to generate such a model is collagenase injection, although this method possesses limitations. We developed an optimized rat model of patellar tendinopathy via the ultrasound-guided injection of collagenase mixed with a thermo-responsive Pluronic hydrogel into the patellar tendon of sixty male Wistar rats. All analyses were carried out at 3, 7, 14, 30, and 60 days post-injury. We confirmed that our rat model reproduced the pathophysiology observed in human patients through analyses of ultrasonography, histology, immunofluorescence, and biomechanical parameters. Tendons that were injured by the injection of the collagenase-Pluronic mixture exhibited a significant increase in the cross-sectional area (p < 0.01), a high degree of tissue disorganization and hypercellularity, significantly strong neovascularization (p < 0.01), important changes in the levels of types I and III collagen expression, and the organization and presence of intra-tendinous calcifications. Decreases in the maximum rupture force and stiffness were also observed. These results demonstrate that our model replicates the key features observed in human patellar tendinopathy. Collagenase is evenly distributed, as the Pluronic hydrogel prevents its leakage and thus, damage to surrounding tissues. Therefore, this model is valuable for testing new treatments for patellar tendinopathy.

The rat Achilles and patellar tendons have similar increases in mechanical properties but become transcriptionally divergent during postnatal development.

The molecular events that drive post-natal tendon development are poorly characterized. In this study, we profiled morphological, mechanical, and transcriptional changes in the rat Achilles and patellar tendon before walking (P7), shortly after onset of walking (P14), and at motor maturity (P28). The Achilles and patellar tendons increased collagen content and mechanical strength similarly throughout post-natal development. However, at P28 the patellar tendon tended to display a higher maximal tensile load (MTL) (P = 0.0524) than the Achilles tendon, but a similar ultimate tensile strength (UTS; load relative to cross-sectional area) probably due to its increased cross-sectional area during development. The tendons started transcriptionally similar, with overlapping PCA clusters at P7 and P14, before becoming transcriptionally distinct at P28. In both tendons, there was an increase in extracellular matrix (ECM) gene expression and a concomitant decrease in cell cycle and mitochondrial gene expression. The transcriptional divergence at P28 suggested that STAT signalling was lower in the patellar tendon where MTL increased the most. Treating engineered human ligaments with the STAT inhibitor itacitinib increased collagen content and MTL. Our results suggest that during post-natal development, cellular resources are initially allocated towards cell proliferation before shifting towards extracellular matrix development following the onset of mechanical load and provide potential targets for improving tendon function. KEY POINTS: Little is known about mechanisms of post-natal tendon growth. We characterized morphological, mechanical, and transcriptional changes that occur before (P7), and early (P14) and late after (P28) rats begin to walk. From P7 to P28, the Achilles tendon increased in length, whereas the patellar tendon increased in cross-sectional area. Mechanical and material properties of the Achilles and patellar tendon increased from P7 to P28. From P7 to P28, the Achilles and patellar tendons increased expression of ECM genes and decreased mitochondrial and cell cycle gene expression. Ribosomal gene expression also significantly decreased in the Achilles and tended to decrease in the patellar tendon. At P28, STAT1 signalling tended to be lower in the patellar tendon which had grown by increasing cross-sectional area and inhibiting STAT activation in vitro improved mechanical properties in engineered human ligaments.

Near infrared spectroscopic evaluation of biochemical and crimp properties of knee joint ligaments and patellar tendon.

Knee ligaments and tendons play an important role in stabilizing and controlling the motions of the knee. Injuries to the ligaments can lead to abnormal mechanical loading of the other supporting tissues (e.g., cartilage and meniscus) and even osteoarthritis. While the condition of knee ligaments can be examined during arthroscopic repair procedures, the arthroscopic evaluation suffers from subjectivity and poor repeatability. Near infrared spectroscopy (NIRS) is capable of non-destructively quantifying the composition and structure of collagen-rich connective tissues, such as articular cartilage and meniscus. Despite the similarities, NIRS-based evaluation of ligament composition has not been previously attempted. In this study, ligaments and patellar tendon of ten bovine stifle joints were measured with NIRS, followed by chemical and histological reference analysis. The relationship between the reference properties of the tissue and NIR spectra was investigated using partial least squares regression. NIRS was found to be sensitive towards the water (R2CV = .65) and collagen (R2CV = .57) contents, while elastin, proteoglycans, and the internal crimp structure remained undetectable. As collagen largely determines the mechanical response of ligaments, we conclude that NIRS demonstrates potential for quantitative evaluation of knee ligaments.

Tendon and motor phenotypes in the Crtap-/- mouse model of recessive osteogenesis imperfecta.

Osteogenesis imperfecta (OI) is characterized by short stature, skeletal deformities, low bone mass, and motor deficits. A subset of OI patients also present with joint hypermobility; however, the role of tendon dysfunction in OI pathogenesis is largely unknown. Using the Crtap-/- mouse model of severe, recessive OI, we found that mutant Achilles and patellar tendons were thinner and weaker with increased collagen cross-links and reduced collagen fibril size at 1- and 4-months compared to wildtype. Patellar tendons from Crtap-/- mice also had altered numbers of CD146+CD200+ and CD146-CD200+ progenitor-like cells at skeletal maturity. RNA-seq analysis of Achilles and patellar tendons from 1-month Crtap-/- mice revealed dysregulation in matrix and tendon marker gene expression concomitant with predicted alterations in TGF-ß, inflammatory, and metabolic signaling. At 4-months, Crtap-/- mice showed increased αSMA, MMP2, and phospho-NFκB staining in the patellar tendon consistent with excess matrix remodeling and tissue inflammation. Finally, a series of behavioral tests showed severe motor impairments and reduced grip strength in 4-month Crtap-/- mice - a phenotype that correlates with the tendon pathology.

Tenascin-C regulates migration of SOX10 tendon stem cells via integrin-α9 for promoting patellar tendon remodeling.

Insufficient attention has been focused on the directional migration of SOX10+ tendon stem cells (STSCs) during tendon remodeling. Here, we investigate whether tenascin-C (TNC) promotes STSC motility and migration. Based on the hypothesis that TNCs induce STSC migration, RNA-sequencing (RNA-seq) was conducted, identifying 2107 differentially expressed genes (DEGs), of which 1272 were up-regulated and 835 down-regulated following treatment with TNC versus the control. The DEGs were principally involved in cell adhesion and cell membrane signal transduction. Highly enriched-related signaling included the PI3K-Akt, focal adhesion, and ECM-receptor interaction pathways. Protein interaction analysis established that TNC was positively correlated with ITGA9 (integrin-α9). Furthermore, TNC activated the phosphorylation levels of FAK and Akt, and knockdown of ITGA9 with siRNA revealed that TNC contributes to STSC migration via the targeting of ITGA9. In addition, in vivo administration of TNC promoted tissue regeneration of injured tendons. In conclusion, TNC regulated the migration of STSCs via ITGA9, thereby promoting the regeneration of tendon injuries.

Impact of plantaris ligamentous tendon.

There are countless morphological variations among the muscles, tendons, ligaments, arteries, veins and nerves of the human body, many of which remain undescribed. Anatomical structures are also subject to evolution, many disappearing and others continually emerging. The main goal of this pilot study was to describe a previously undetected anatomical structure, the plantaris ligamentous tendon, and to determine its frequency and histology. Twenty-two lower limbs from 11 adult cadavers (11 left, and 11 right) fixed in 10% formalin were examined. The mean age of the cadavers at death was 60.1 years (range 38-85). The group comprised six women and five men from a Central European population. All anatomical dissections of the leg and foot area accorded with the pre-established protocol. Among the 22 lower limbs, the PLT was present in 16 (72.7%) and absent in six (27.3%). It originated as a strong fan-shaped ligamentous tendon from the superior part of the plantaris muscle, the posterior surface of the femur and the lateral aspect of the knee joint capsule. It inserted to the ilio-tibial band. Histologically, a tendon and ligament were observed extending parallel to each other. A new anatomical structure has been found, for which the name plantaris ligamentous tendon is proposed. It occurs around the popliteal region between the plantaris muscle, the posterior surface of the femur, and the ilio-tibial band.

Cruciate ligament, patellar tendon, and patella formation involves differential cellular sources and dynamics as joint cavitation proceeds.

BACKGROUND: Cruciate ligament (CL) and patellar tendon (PT) are important elements of the knee joint, uniting femur, patella, and tibia into a single functional unit. So far, knowledge on the developmental mechanism of CL, PT, and patella falls far behind other skeletal tissues. RESULTS: Here, employing various lineage tracing strategies we investigate the cellular sources and dynamics that drive CL, PT, and patella formation during mouse embryonic development. We show that Gdf5 and Gli1 are generally expressed in the same cell population that only contributes to CL, but not PT or patella development. In addition, Col2 is expressed in two independent cell populations before and after joint cavitation, where the former contributes to the CL and the dorsal part of the PT and the latter contributes to the patella. Moreover, Prrx1 is always expressed in CL and PT progenitors, but not patella progenitors where it is switched off after joint cavitation. Finally, we reveal that patella development employs different cellular dynamics before and after joint cavitation. CONCLUSIONS: Our findings delineate the expression changes of several skeletogenesis-related genes before and after joint cavitation, and provide an indication on the cellular dynamics underlying ligament, tendon, and sesamoid bone formation during embryogenesis.

Electrospun PCL/Gel-aligned scaffolds enhance the biomechanical strength in tendon repair.

Tendons can transmit mechanical force from muscles to bones for movement. However, the mechanical strength of tendons is compromised after surgery, thus causing a high rate of tendon retear. Hence, the design and preparation of biodegradable materials with excellent mechanical properties have become an urgent demand for sports medicine. In this study, biomimetic polycaprolactone (PCL)/gelatin (Gel)-aligned scaffolds were fabricated for the mechanical restoration of the injured tendon in a rabbit model. The diameter of nanofibers was about 427.82 ± 56.99 nm, which was approximate to that of the native collagen fibrils; the directional consistency of the nanofibers in PCL/Gel-aligned scaffolds reached 77.33 ± 3.22%, which were ultrastructurally biomimetic. Compared to the observations for the control group, the in vitro mechanical results showed that the PCL/Gel-aligned scaffolds (P/G-A) were anisotropic in terms of failure load, tensile strength, and Young's modulus. After verifying their good cytocompatibility, the scaffolds were implanted into the rabbit patellar tendon in situ. The biomechanical properties of the repaired tendon in P/G-A reached 343.97 ± 65.30 N in failure load, 85.99 ± 16.33 MPa in tensile strength, 590.84 ± 201.87 MPa in Young's modulus, and 171.29 ± 61.50 N mm-1 in stiffness in vivo at 8 weeks post operation. In a word, our results demonstrated that P/G-A could support the regenerated tissue of injured patellar tendons to restore the biomechanical strength in a rabbit model. This suggested that the PCL/Gel-aligned scaffolds can pave a promising way to improve the healing of injured tendons in the clinic in the future.

Autologous Freeze-Dried, Platelet-Rich Plasma Carrying Icariin Enhances Bone-Tendon Healing in a Rabbit Model.

BACKGROUND: Tendon-bone interface (TBI) injuries are common in sports activities. Owing to the limited regenerative ability of the TBI, its functional healing remains a difficulty in clinical practice. Icariin (ICA) provides strong stimulation for osteogenesis. Platelet-rich plasma (PRP) can be used as a carrier for bioactive molecules, although its ability to provide sustained release for such molecules needs improvement. HYPOTHESIS: Freeze-dried PRP (FD-PRP) as a carrier for ICA can provide sustained release of ICA into the tendon-bone (T-B) healing site, thus accelerating T-B healing. STUDY DESIGN: Controlled laboratory study. METHODS: A total of 84 New Zealand rabbits with partial patellectomy in the hindlimb were randomly allocated into 3 different treatments: ICA incorporated with FD-PRP (ICA/FD-PRP), FD-PRP alone (FD-PRP), or saline control (CTL). The rabbit patella-patellar tendon (PP) interfaces were postoperatively harvested at postoperative week 8 or 16 for gross, radiological, histological, and mechanical evaluations. RESULTS: Our results showed that FD-PRP can act as a carrier for sustained release of ICA into the T-B healing site. Macroscopically, no signs of infection or osteoarthritis were shown in the regenerated PP interfaces, and the area of cartilaginous metaplasia in the FD-PRP and ICA/FD-PRP groups at postoperative week 16 was significantly larger than that of the CTL group (P < .05 for all). Radiologically, micro-computed tomography showed that new bone which formed at the healing site in the ICA/FD-PRP group was significantly increased, remodeled, and mineralized in comparison with the CTL group (P < .05 for all). Histologically, the ICA/FD-PRP group exhibited a significant native PP interface, as shown by the enlargement and remodeling of new bone, well-organized collagen fibers, and robust production of proteoglycans in the regenerated fibrocartilage. The mechanical strength of the regenerated PP interface was significantly improved in the ICA/FD-PRP group. Significantly higher failure load and stiffness were shown in the ICA/FD-PRP group compared with the CTL and FD-PRP groups, respectively (P < .05 for all). CONCLUSION: FD-PRP is a suitable sustained-release carrier for ICA, and ICA/FD-PRP can provide sustained release of ICA into the T-B healing site, thus effectively accelerating T-B healing. CLINICAL RELEVANCE: Findings of this study demonstrate the feasibility of using FD-PRP as a carrier for ICA to improve T-B healing and provide a foundation for future clinical application.

Investigating circadian clock gene expression in human tendon biopsies from acute exercise and immobilization studies.

PURPOSE: The discovery of musculoskeletal tissues, including muscle, tendons, and cartilage, as peripheral circadian clocks strongly implicates their role in tissue-specific homeostasis. Age-related dampening and misalignment of the tendon circadian rhythm and its outputs may be responsible for the decline in tendon homeostasis. It is unknown which entrainment signals are responsible for the synchronization of the tendon clock to the light-dark cycle. METHODS: We sought to examine any changes in the expression levels of core clock genes (BMAL1, CLOCK, PER2, CRY1, and NR1D1) in healthy human patellar tendon biopsies obtained from three different intervention studies: increased physical activity (leg kicks for 1 h) in young, reduced activity (2 weeks immobilization of one leg) in young, and in old tendons. RESULTS: The expression level of clock genes in human tendon in vivo was very low and a high variation between individuals was found. We were thus unable to detect any differences in core clock gene expression neither after acute exercise nor immobilization. CONCLUSIONS: We are unable to find evidence for an effect of exercise or immobilization on circadian clock gene expression in human tendon samples.

A potential new role for myofibroblasts in remodeling of sub-rupture fatigue tendon injuries by exercise.

Tendons are ineffective at repairing sub-rupture fatigue injuries. Accordingly, we evaluated whether an exercise protocol that we have previously found to decrease structural damage kinks in fatigue damaged tendons, leads to improvement in mechanical properties. We hypothesized that exercise that promotes repair of fatigue damage will decrease apoptosis and increase the population of myofibroblasts. Rat patellar tendons underwent in vivo fatigue loading for 500 or 7200 cycles. Animals resumed cage activity for 2-weeks, then either remained cage active or began treadmill running until sacrifice at 4- or 10-weeks post-fatigue loading. Exercise following fatigue damage increased the stiffness back towards naïve levels, decreased apoptosis and increased the population of myofibroblasts. Next, proteins associated with inhibition of apoptosis (Collagen VI) or activation of myofibroblast (pSmad 2/3, fibrillin, integrin subunits αV and α5) were evaluated. Data suggests that collagen VI may not be integral to inhibition of apoptosis in this context. Exercise increased pSmad 2/3 and fibrillin in the insertion region for the 7200-cycles group. In addition, exercise decreased integrin αV and increased integrin α5 in fatigue damaged tendons. Data suggests that a decrease in apoptosis and an increase in population of myofibroblasts may be integral to remodeling of fatigue damaged tendons.

Streptozotocin-induced diabetes alters transcription of multiple genes necessary for extracellular matrix remodeling in rat patellar tendon.

OVERVIEW: Tendon collagen fibril degradation is commonly seen in tendons of diabetics, but the mechanisms responsible for these changes remain to be elucidated. We have demonstrated that streptozotocin (STZ)-induced diabetes increases tendon cell proliferation and collagen content. In the present study, we evaluated that impact of STZ-induced diabetes on mRNA transcripts involved with collagen fibril organization, extracellular matrix (ECM) remodeling, apoptosis, and proliferation. MATERIALS AND METHODS: Rats were divided into four groups: nondiabetic (control, n = 9), 1 week (acute, n = 8) or 10 weeks of diabetes (chronic, n = 7), and 10 weeks of diabetes with insulin (insulin, n = 8). RNA was isolated from the patellar tendon for determination of mRNA transcripts using droplet digital PCR (ddPCR). RESULTS: Transcripts for Col1a1, Col3a1, Mmp2, Timp1, Scx, Tnmd, Casp3, Casp8, and Ager were lower in acute relative to control and insulin rats (p ≤ 0.05). With the exception of Scx, transcripts for Col1a1, Col3a1, Mmp2, Timp1, Tnmd, Casp3, Casp8, and Ager were also lower in chronic when compared to control (p < 0.05). Transcripts for Col1a1, Col3a1, Mmp2, Timp1, Tnmd, Casp3, Casp8, and Ager were not different between control and insulin (p > 0.05). Transcripts for Dcn, Mmp1a, Mmp9, Pcna, Tgfbr3, Ptgs2, Ptger2, Ptges, and iNos were not altered by diabetes or insulin (p > 0.05). CONCLUSION: Our findings indicated that STZ-induced diabetes results in rapid and large changes in the expression of several genes that are key to ECM remodeling, maintenance, and maturation.

Enhanced biological fixation of ligaments to bone tissues utilizing chitin fabrics.

In ligament reconstruction involving anterior cruciate ligament surgery, biological fixation between the transferred ligament and bone tissue is critical for achieving successful outcomes. Here, we administered chitin fabrics into the bone tunnels and evaluated their efficacy in promoting biological fixation. An animal model on the rat's patellar ligament was employed. First, bone tunnels were created in the lateral condyle of the femur. The ligament was then separated from the tibial tuberosity, and half was inserted into the tunnel and fixed with the use of end button. Animals in the experimental group were treated with microfiber nonwoven chitin fabric, whereas control animals received no treatment. Specimens were collected at 2, 4, and 6 weeks after surgery, and the fixation strength was measured by mechanical tests. Histological sections were prepared from samples prepared 4 weeks after surgery, and the diameter of bone tunnel and the width ratio of collagenous tissue in the bone tunnel were measured. Administration of chitin significantly increased the mean fixation strength at 4 and 6 weeks after surgery. Furthermore, chitin also promoted bone formation in the bone tunnel and increased the density of collagen fibers. Thus, microfiber nonwoven chitin fabric enhanced the biological fixation of the ligament to the bone tissue. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2355-2360, 2018.

Influence of the Infrapatellar Fat Pad Resection during Total Knee Arthroplasty: A Systematic Review and Meta-Analysis.

BACKGROUND: To enhance surgical exposure, resection of the infrapatellar fat pad (IPFP) is usually a routine procedure in total knee arthroplasty (TKA). However, there is conflicting evidence regarding whether IPFP resection during TKA impairs clinical outcome. We performed a systematic review and meta-analysis to clarify the influence of IPFP resection on primary TKA. METHODS: Embase, PubMed, and the Cochrane Library were systematically searched up to August 2016 to identify relevant studies. All clinical studies comparing IPFP resection (IPFP-R) and IPFP preservation (IPFP-P) in patients undergoing primary TKA were obtained. The meta-analysis was performed with Revman 5.3 and STATA 12.0 software. The weighted mean was estimated by using random effects (RE) models with 95% CIs, heterogeneity was assessed using the H statistic and the inconsistency index (I2). RESULTS: Seven studies involving 2,734 patients (3,258 knees) were included. IPFP resection trended to increase the incidence of postoperative anterior knee pain within 2 months postoperatively, compared with patients in whom the IPFP was preserved (odds ratio [OR]s 2.12[0.95, 4.73], p = 0.07). An increased incidence of anterior knee pain was observed in the IPFP resection group > 12 months postoperatively, but the difference was not significant (OR, 3.69 [0.81, 16.82], p = 0.07). In addition, a trend towards more shortening of the patellar tendon was also observed in the IPFP-R group. No significant results were found regarding postoperative knee function. CONCLUSION: These results suggest that preserving the IPFP may be superior to IPFP resection in patients undergoing primary TKA, due to the relatively lower rate of anterior knee pain after short-term follow-up.

Changes in Knee Laxity and Relaxin Receptor Isoforms Expression (RXFP1/RXFP2) in the Knee throughout Estrous Cycle Phases in Rodents.

The changes in knee laxity and relaxin receptor expression at different phases of rodent estrous cycle are not known. Here, changes in the parameter were investigated in rats at different phases of the estrous cycle. Estrous cycle phases of intact female rats were determined by cytological examination of the vaginal smear. Following phase identification, blood was collected for serum hormone analyses. Knee passive range of motion (ROM) was determined by using a digital miniature goniometer. The animals were then sacrificed and patellar tendon, collateral ligaments and hamstring muscles were harvested for relaxin/insulin-like family peptide receptor 1 and 2 (RXFP1/RXFP2) analyses. Knee passive ROM was the highest at proestrus followed by diestrus and the lowest at estrus. Estrogen level was the highest at proestrus while progesterone and relaxin levels were the highest at diestrus. A strong correlation was observed between relaxin and progesterone levels. At proestrus, expression of RXFP1 and RXFP2 proteins and mRNAs were the highest at proestrus followed by diestrus and estrus. The finding shows that higher level of progesterone and relaxin in diestrus might be responsible for higher laxity of knee joint in rats.

Changes in water content in response to an acute bout of eccentric loading in a patellar tendon with a history of tendinopathy: A case report.

OBJECTIVE: This case-based report assessed resting water content and exercise-driven water exchange within a tendon with a history of tendinopathy and compared the response to that of a healthy uninvolved tendon. DESIGN: Case Report. SETTING: University imaging center. PARTICIPANT: The participant was a 27-year-old female basketball player 39 months following knee trauma. Patellar tendinopathy developed 12 months after the injury episode and was treated with eccentric exercises. Eighteen months from the beginning of the first eccentric training bout, the participant reported full resolution of symptoms and returned to her pre-injury sport participation without symptoms. INTERVENTION: Eccentric decline squat exercise. MAIN OUTCOME MEASURES: Tendon water content obtained from magnetic resonance imaging (MRI). RESULTS: MRI acquired 39 months post-injury demonstrated increased resting water content of the involved tendon (involved: 91.1% vs. uninvolved: 84.6%). Immediately after the eccentric squat maneuver, water content decreased on both involved and uninvolved tendons (involved: 89.5% vs. uninvolved: 83.3%). CONCLUSIONS: Elevated resting water content of the involved tendon found in this report may be indicative of reduced tendon stiffness. A similar amount of water content reduction was observed on both sides following mechanical loading, suggesting that the involved tendon may respond to the eccentric exercise similarly to the uninvolved tendon. Future investigations are needed to study the relationships among tendon water exchanges, mechanical properties, patient symptoms, and tissue injuries.

Stepwise Differentiation of Mesenchymal Stem Cells Augments Tendon-Like Tissue Formation and Defect Repair In Vivo.

UNLABELLED: : Tendon injuries are common and present a clinical challenge, as they often respond poorly to treatment and result in long-term functional impairment. Inferior tendon healing responses are mainly attributed to insufficient or failed tenogenesis. The main objective of this study was to establish an efficient approach to induce tenogenesis of bone marrow-derived mesenchymal stem cells (BMSCs), which are the most common seed cells in tendon tissue engineering. First, representative reported tenogenic growth factors were used as media supplementation to induce BMSC differentiation, and the expression of teno-lineage transcription factors and matrix proteins was compared. We found that transforming growth factor (TGF)-ß1 significantly induced teno-lineage-specific gene scleraxis expression and collagen production. TGF-ß1 combined with connective tissue growth factor (CTGF) elevated tenomodulin and Egr1 expression at day 7. Hence, a stepwise tenogenic differentiation approach was established by first using TGF-ß1 stimulation, followed by combination with CTGF for another 7 days. Gene expression analysis showed that this stepwise protocol initiated and maintained highly efficient tenogenesis of BMSCs. Finally, regarding in situ rat patellar tendon repair, tendons treated with induced tenogenic BMSCs had better structural and mechanical properties than those of the control group, as evidenced by histological scoring, collagen I and tenomodulin immunohistochemical staining, and tendon mechanical testing. Collectively, these findings demonstrate a reliable and practical strategy of inducing tenogenesis of BMSCs for tendon regeneration and may enhance the effectiveness of cell therapy in treating tendon disorders. SIGNIFICANCE: The present study investigated the efficiency of representative tenogenic factors on mesenchymal stem cells' tenogenic differentiation and established an optimized stepwise tenogenic differentiation approach to commit tendon lineage differentiation for functional tissue regeneration. The reliable tenogenic differentiation approach for stem cells not only serves as a platform for further studies of underlying molecular mechanisms but also can be used to enhance cell therapy outcome in treating tendon disorders and develop novel therapeutics for tendon injury.

Proteomic differences between native and tissue-engineered tendon and ligament.

Tendons and ligaments (T/Ls) play key roles in the musculoskeletal system, but they are susceptible to traumatic or age-related rupture, leading to severe morbidity as well as increased susceptibility to degenerative joint diseases such as osteoarthritis. Tissue engineering represents an attractive therapeutic approach to treating T/L injury but it is hampered by our poor understanding of the defining characteristics of the two tissues. The present study aimed to determine differences in the proteomic profile between native T/Ls and tissue engineered (TE) T/L constructs. The canine long digital extensor tendon and anterior cruciate ligament were analyzed along with 3D TE fibrin-based constructs created from their cells. Native tendon and ligament differed in their content of key structural proteins, with the ligament being more abundant in fibrocartilaginous proteins. 3D T/L TE constructs contained less extracellular matrix (ECM) proteins and had a greater proportion of cellular-associated proteins than native tissue, corresponding to their low collagen and high DNA content. Constructs were able to recapitulate native T/L tissue characteristics particularly with regard to ECM proteins. However, 3D T/L TE constructs had similar ECM and cellular protein compositions indicating that cell source may not be an important factor for T/L tissue engineering.

Moderate treadmill running exercise prior to tendon injury enhances wound healing in aging rats.

The effect of exercise on wound healing in aging tendon was tested using a rat moderate treadmill running (MTR) model. The rats were divided into an MTR group that ran on a treadmill for 4 weeks and a control group that remained in cages. After MTR, a window defect was created in the patellar tendons of all rats and wound healing was analyzed. We found that MTR accelerated wound healing by promoting quicker closure of wounds, improving the organization of collagen fibers, and decreasing senescent cells in the wounded tendons when compared to the cage control. MTR also lowered vascularization, increased the numbers of tendon stem/progenitor cells (TSCs) and TSC proliferation than the control. Besides, MTR significantly increased the expression of stem cell markers, OCT-4 and Nanog, and tenocyte genes, Collagen I, Collagen III and tenomodulin, and down-regulated PPAR-γ, Collagen II and Runx-2 (non-tenocyte genes). These findings indicated that moderate exercise enhances healing of injuries in aging tendons through TSC based mechanisms, through which exercise regulates beneficial effects in tendons. This study reveals that appropriate exercise may be used in clinics to enhance tendon healing in aging patients.

Local trauma in human patellar tendon leads to widespread changes in the tendon gene expression.

Low cellular activity and slow tissue turnover in human tendon may prolong resolution of tendinopathy. This may be stimulated by moderate localized traumas such as needle penetrations, but whether this results in a widespread cellular response in tendons is unknown. In an initial hypothesis-generating study, a trauma-induced tendon cell activity (increased total RNA and collagen I mRNA) was observed after repeated patellar tendon biopsies in young men. In a subsequent controlled study, 25 young men were treated with two 0.8-mm-diameter needle penetrations [n = 13, needle-group (NG)] or one 2.1-mm-diameter needle biopsy [n = 12, biopsy-group (BG)] in one patellar tendon. Four weeks later biopsies were taken from treated (5 mm lateral from trauma site) and contralateral tendons for analyses of RNA content (ribogreen assay), DNA content (PCR based), and gene expression for relevant target genes (Real-time RT-PCR) (NG, n = 11 and BG, n = 8). Intervention increased RNA content, and mRNA expression of collagen I and III and TGF-ß1 (P < 0.05), with biopsy treatment having greatest effect (tendency for RNA and collagen I). Results for DNA content were inconclusive, and no changes were detected in expression of insulin-like growth factor-I, connective tissue growth factor, scleraxis, decorin, fibromodulin, tenascin-C, tenomodulin, VEGFa, CD68, IL-6, MMP12, and MMP13. In conclusion, a moderate trauma to a healthy human tendon (e.g., biopsy sampling) results in a widespread upregulation of tendon cell activity and their matrix protein expression. The findings have implications for design of studies on human tendon and may provide perspectives in future treatment strategies in tendinopathy.