Differential MMP 1 and MMP 13 expression in proliferation and ligamentization phases of graft remodeling in anterior cruciate ligament reconstruction.

Purpose: Graft remodeling in anterior cruciate ligament reconstruction (ACLR) demonstrates three distinct phases: necrosis, proliferation and ligamentization. Biological enhancement involves modulating these processes, but the cellular activities related to extracellular matrix remodeling have not been investigated. We hypothesized that changes in matrix metalloproteinases (MMPs) 1 and 13 expression are involved in the transition of proliferation phase to ligamentization phase of graft remodeling.Materials and methods: Thirty-three rats underwent ACLR. Tendon grafts were harvested at week 1 (necrosis), 2 (proliferation), or 12 (ligamentization) post-operation for histological examination (n = 3), or for isolation of graft-derived cells (n = 8) for flow cytometry, proliferation assay, cell invasion assay, measurement of gene expression related to matrix remodeling (Col1A1, Col3A1, MMP1, tissue inhibitor of marix metalloproteinase 1 (TIMP1), and MMP13) and total MMP activities.Results: Increased cellularity in tendon graft was contributed by active cell proliferation and migration at week 2 post-operation, while decreased cellularity were paralleled by increased apoptosis at week 12. All genes measured (Col1A1, Col3A1, MMP1, TIMP1, and MMP13) increased significantly in week 2 cells compared to week 1 cells. MMP1 expression subsided at week 12, while MMP13 expression kept increasing till 12 weeks post-operation. Total MMP activities was 3-fold higher in cultured graft-derived cells from week 2 as compared to cells from week 12. Two distinct processes of graft remodeling were identified, characterized by increased MMP1 expression with cell proliferation and increased MMP13 expression with cell apoptosis.Conclusions: Unfavorable matrix remodeling during the proliferation phase is found with increased MMP1, while remodeling leading to ligamentization is associated with increased MMP13 expression.

A systematic review of inflammatory cells and markers in human tendinopathy.

BACKGROUND: This article systematically reviews the current evidence regarding inflammation in Tendinopathy with the aim to increase understanding of a potential common pathophysiology. METHODS: Following the PRISMA statements, the terms: (tendinopathy OR (tendons AND rupture)) AND (inflammation OR (inflammation AND cells) OR immune system OR inflammation mediators OR bacteria) were used. One thousand four hundred thirty-one articles were identified which was screened down to 53. RESULTS: 39/53 studies mentioned inflammatory cells but had contradicting conclusions. Macrophages were the most common cell type and inflammatory markers were detectable in all the articles which measure them. CONCLUSIONS: The included studies show different conclusions, but this heterogeneity is not unexpected since the clinical criteria of 'tendinopathy' encompass a huge clinical spectrum. Different 'tendinopathy' conditions may have different pathophysiology, and even the same clinical condition may be at different disease stages during sampling, which can alter the histological and biochemical picture. Control specimen sampling was suboptimal since the healthy areas of the pathological-tendon may actually be sub-clinically diseased, as could the contralateral tendon in the same subject. Detection of inflammatory cells is most sensitive using immunohistochemistry targeting the cluster of differentiation markers, especially when compared to the conventional haematoxylin and eosin staining methods. The identified inflammatory cell types favour a chronic inflammatory process; which suggests a persistent stimulus. This means NSAID and glucocorticoids may be useful since they suppress inflammation, but it is noted that they may hinder tendon healing and cause long term problems. This systematic review demonstrates a diversity of data and conclusions in regard to inflammation as part of the pathogenesis of Tendinopathy, ranging from ongoing or chronic inflammation to non-inflammatory degeneration and chronic infection. Whilst various inflammatory markers are present in two thirds of the reviewed articles, the heterogenicity of data and lack of comparable studies means we cannot conclude a common pathophysiology from this systematic review.

Long noncoding RNA H19 accelerates tenogenic differentiation and promotes tendon healing through targeting miR-29b-3p and activating TGF-ß1 signaling.

Tendon injures are common orthopedic conditions, but tendon development and the pathogenesis of tendon injures, such as tendinopathy, remain largely unknown and have limited the development of clinical therapy. Studies on tenogenic differentiation at the molecular level may help in developing novel therapeutic strategies. As novel regulators, long noncoding RNAs (lncRNAs) have been found to have widespread biological functions, and emerging evidence demonstrates that lncRNAs may play important regulatory roles in cell differentiation and tissue regeneration. In this study, we found that lncRNA H19 stimulated tenogenesis of human tendon-derived stem cells. Stable overexpression of H19 significantly accelerated TGF-ß1-induced tenogenic differentiation in vitro and accelerated tendon healing in a mouse tendon defect model. H19 directly targeted miR-29b-3p, which is considered to be a negative regulator of tenogenesis. Furthermore, miR-29b-3p directly suppressed the expression of TGF-ß1 and type I collagen, thereby forming a novel regulatory feedback loop between H19 and TGF-ß1 to mediate tenogenic differentiation. Our study demonstrated that H19 promotes tenogenic differentiation both in vitro and in vivo by targeting miR-29b-3p and activating TGF-ß1 signaling. Regulation of the TGF-ß1/H19/miR-29b-3p regulatory loop may be a new strategy for treating tendon injury.-Lu, Y.-F., Liu, Y., Fu, W.-M., Xu, J., Wang, B., Sun, Y.-X., Wu, T.-Y., Xu, L.-L, Chan, K.-M., Zhang, J.-F., Li, G. Long noncoding RNA H19 accelerates tenogenic differentiation and promotes tendon healing through targeting miR-29b-3p and activating TGF-ß1 signaling.

Tenogenic differentiation of mesenchymal stem cells and noncoding RNA: From bench to bedside.

Tendon is a critical unit of musculoskeletal system that connects muscle to bone to control bone movement. More population participate in physical activities, tendon injuries, such as acute tendon rupture and tendinopathy due to overuse, are common causing unbearable pain and disability. However, the process of tendon development and the pathogenesis of tendinopathy are not well defined, limiting the development of clinical therapy for tendon injuries. Studying the tendon differentiation control pathways may help to develop novel therapeutic strategies. This review summarized the novel molecular and cellular events in tendon development and highlighted the clinical application potential of non-coding RNAs and tendon-derived stem cells in gene and cell therapy for tendon injuries, which may bring insights into research and new therapy for tendon disorders.

Tendon-derived stem cells undergo spontaneous tenogenic differentiation.

Tendon-derived stem cell (TDSC) is a subpopulation of residing stem cells within the intact tendon tissues, with the capacities of self-renewal, clonogenicity, and three-lineage differentiation. Compared with bone marrow derived mesenchymal stem cells (BMSCs), TDSCs are superior for tendon injuries repair as they remain some tendon tissue-specific differentiation properties. In the present study, TDSC was found to undergo spontaneous tenogenic differentiation in which the expression of tenogenic markers were increased while the expression of stemness markers decreased with time in TDSCs culture (without tenogenic induction medium). The further collagen synthesis ability was correspondingly increased during this process. After a longer period of culture, the monolayer of TDSCs formed a "3D" layers with rich extracellular matrices of typical tendon tissues. In addition, the key tenogenic transcription factors, such as Scx, Mkx, Egr1 and Eya1 were all up-regulated in this process. Finally, we compared the spontaneous tenogenic differentiation with TGF-ß1-induced tenogenic differentiation of TDSCs, and the results showed that the spontaneous tenogenic differentiation of TDSCs was general character of TDSCs, similar to but weaker than the effect of TDSCs under tenogenic induction. Taken together, the present study identified that TDSCs had the potential of spontaneous tenogenic differentiation, which may be a better cell source for the treatment of tendon injury.

The reliability and validity of a video-based method for assessing hamstring strength in football players.

BACKGROUND/OBJECTIVE: Evaluating hamstring strength by isokinetic dynamometry is limited by various practical issues such as time and cost. A video-based Nordic hamstring exercise is introduced as an alternative option. The aims of this study are to evaluate 1.) the between-session reliability and 2.) concurrent validity of the testing method compared to a standardized isokinetic dynamometry. METHODS: Thirty male elite footballers were recruited for the study. From the Nordic hamstring exercise, the video-analysis-determined Nordic break-point angles where the participant could no longer withstand the force of the fall (eccentric mode) and the number of seconds that the player could hold at 30° forward flexion angle (isometric mode) were measured. Intra-class correlation coefficients for between-session reliability, Pearson r correlations between the current method and isokinetic dynamometry were calculated. RESULTS: The reliability of the eccentric mode was moderate (ICC (2,1) = 0.82) while that of isometric mode was poor (ICC (2,1) = 0.57). The Nordic break-point angle of the eccentric mode significantly correlated with the concentric and eccentric hamstring peak torque (r = 0.48 and 0.58, p < 0.001), while the isometric was not (r = 0.02 - 0.07, p > 0.05). CONCLUSION: The eccentric mode of the video-based hamstring strength test was a moderately reliable and valid method to measure the eccentric hamstring strength in elite football players.

Motion Task Selection for Kinematic Evaluation After Anterior Cruciate Ligament Reconstruction: A Systematic Review.

PURPOSE: To examine the different motion tasks and the protocols used to objectively quantify dynamic stability in terms of knee kinematics at different stages of anterior cruciate ligament reconstruction (ACLR) recovery. METHODS: A systematic search was done using OVID in Embase, Cochrane Central Register of Controlled Trials, Medline, PsychINFO, and AMED. A combination of the following keywords and their variations were used: anterior cruciate ligament, motion tasks (e.g., jump, hop, gait), and stability. The inclusion criteria were as follows: (1) ACLR subjects were recruited, (2) at least 1 motion task was performed and kinematics data were recorded, and (3) uninjured subjects or the contralateral uninjured limbs were included as a control group. Exclusion criteria were as follows: (1) non-English language publications, (2) retrospective studies and review articles, (3) animal studies, and (4) cadaveric studies. RESULTS: The search returned 2,195 studies, and 56 were included in this review according to the criteria. A total of 1,086 ACLR subjects were included. Pivoting, landing, walking, running, stair negotiation, and squats were assessed using optoelectronic motion capture, electrogoniometry, or video-radiography. CONCLUSIONS: The appropriate selection of motion tasks is an integral factor in dynamic stability testing as it evokes different kinematic outcomes in relation to the different stages of ACLR recovery. Stair negotiation and landing tasks are best performed during the early stages of recovery, and landing and pivoting are recommended 6 months after ACLR surgery. LEVEL OF EVIDENCE: Level II, systematic review of Level I and II studies.

Immortalized human fetal bone marrow-derived mesenchymal stromal cell expressing suicide gene for anti-tumor therapy in vitro and in vivo.

BACKGROUND AIMS: Cancer is one of the greatest health challenges facing the world today with >10 million new cases of cancer every year. The self-renewal, tumor-homing ability and low immunogenicity of mesenchymal stromal cells (MSCs) make them potential delivery candidates for suicide genes for anti-tumor therapy. However, unstable supply and short life span of adult MSCs in vitro have limited this therapeutic potential. In this study, we aimed to evaluate if immortalization of human fetal bone marrow-derived mesenchymal stromal cells by simian virus 40 (SV40-hfBMSCs) could be a stable source of MSCs for clinical application of suicide gene therapy. METHODS AND RESULTS: Transduction of SV40 and herpes simplex virus thymidine kinase-IRES-green fluorescent protein (TK-GFP) did not cause significant change in the stem cell properties of hfBMSCs. The anti-tumor effect of SV40-TK-hfBMSCs in the presence of the prodrug ganciclovir was demonstrated in vitro and in nude mice bearing human prostate cancer cells, DU145 and PC3, which had been transduced with luciferase and GFP for imaging evaluation by an in vivo live imaging system (IVIS 200 imaging system; Caliper Life Sciences). Repeated injection of low doses (1 × 10(6) cells/kg) of SV40-TK-hfBMSCs was as effective as previously reported and did not cause observable harmful side effects in multiple organs. Mixed lymphocyte reaction showed that SV40-TK-hfBMSCs did not induce significant proliferation of lymphocytes isolated from healthy adults. CONCLUSIONS: Taken together, immortalized hfBMSCs represent a reliable and safe source of MSCs for further clinical translational study.

Effect of graft tensioning on mechanical restoration in a rat model of anterior cruciate ligament reconstruction using free tendon graft.

PURPOSE: Initial graft tensioning is important in anterior cruciate ligament reconstruction (ACLR), but its effect on graft healing is still not clear. Since all previous animal studies on graft tensioning used bone-patellar tendon-bone, this study aimed to investigate the effect of initial graft tensioning on ACLR using tendon graft. METHODS: Fifty-five Sprague-Dawley rats underwent ACLR using flexor digitorum longus tendon graft. A constant force of 2 or 4 N was applied during graft fixation. At 0, 2, and 6 weeks, knee samples were harvested (n = 6) for static knee laxity test and graft pull-out test. Histological examination was performed at 2 and 6 weeks post-injury (n = 4). RESULTS: At time zero, knee laxity was restored by ACLR with 2 or 4 N tensioning as compared to ACL-deficient group (p < 0.001), and the 4 N group exhibited a better restoration as compared to 2 N group (p = 0.031). At week 2 post-operation, the 4 N group still exhibited a better restoration in knee laxity (p = 0.001) and knee stiffness (p = 0.002) than the 2 N group; the graft pull-out force (p = 0.032) and stiffness (p = 0.010) were also higher. At week 6 post-operation, there was no significant difference between the 2 and 4 N group in knee laxity and graft pull-out strength. Histological examination showed that the beneficial effect of higher initial graft tension may be contributed by maintenance of graft integrity at mid-substance and reduction in adverse peri-graft bone changes in the femoral tunnel region. CONCLUSIONS: A higher initial graft tension favours the restoration of knee laxity and promotes graft healing in ACLR using free tendon graft in the rat model.

Biomechanical techniques to evaluate tibial rotation. A systematic review.

PURPOSE: This article systematically reviewed the biomechanical techniques to quantify tibial rotation, for an overview of how to choose a suitable technique for specific clinical application. METHODS: A systematic search was conducted and finally 110 articles were included in this study. The articles were categorized by the conditions of how the knee was examined: external load application, physical examination and dynamic task. RESULTS: The results showed that two-thirds of the included studies measured tibial rotation under external load application, of which over 80% of the experiments employed a cadaveric model. The common techniques used included direct displacement measurement, motion sensor, optical tracking system and universal force moment sensor. Intra-operative navigation system was used to document tibial rotation when the knee was examined by clinical tests. For dynamic assessment of knee rotational stability, motion analysis with skin reflective markers was frequently used although this technique is less accurate due to the skin movement when compared with radiographic measurement. CONCLUSION: This study reports various biomechanical measurement techniques to quantify tibial rotation in the literatures. To choose a suitable measurement technique for a specific clinical application, it is suggested to quantify the effectiveness of a new designed surgical technique by using a cadaveric model before applying to living human subjects for intra-operative evaluation or long-time functional stability assessment. Attention should also be paid on the study's purpose, whether to employ a cadaveric model and the way of stress applied to the knee. LEVEL OF EVIDENCE: IV.

Expression of chondro-osteogenic BMPs in clinical samples of patellar tendinopathy.

PURPOSE: The pathogenesis of patellar tendinopathy remains unclear. Expression of BMP-2/-4/-7 was reported in an ossified failed tendon healing animal model of patellar tendinopathy. This study aimed to investigate the expression of these chondro-osteogenic BMPs in clinical samples of patellar tendinopathy. METHODS: Patellar tendon samples were collected from 16 consecutive patients with patellar tendinopathy and 16 consecutive controls undergoing anterior cruciate ligament reconstruction with bone-patellar tendon-bone autograft in the authors' hospital after getting their consent. The expression of BMP-2/-4/-7 was examined in all samples using immunohistochemistry. Ossification observed in two tendinopathy samples was characterized by histology, alizarin red S staining, alcian blue staining, TRAP staining and immunohistochemical staining of Sox9, osteopontin (OPN) and osteocalcin (OCN). RESULTS: Regions of hypo- and hyper-cellularity and vascularity, with loss of crimp structure of collagen matrix, were observed in patellar tendinopathy samples. Round cells and in some cases, cells with typical chondrocyte phenotype were observed. For the ossified tendinopathy samples with positive alizarin red S staining, OPN-positive and Sox9-positive chondrocyte-like cells in alcian blue-stained extracellular matrix, OCN-positive osteoblast-like cells and TRAP-positive multi-nucleated cells were observed around the ossified deposits. No expression of BMP-2/-4/-7 was observed in healthy patellar tendons. However, the expression of BMP-2/-4/-7 was observed in all patellar tendinopathy samples with or without ossification. CONCLUSIONS: Clinical samples of patellar tendinopathy showed ectopic expression of BMP-2/-4/-7. This was not evident in control samples from healthy patellar tendons. LEVEL OF EVIDENCE: Prognostic studies, Level III.

Higher BMP receptor expression and BMP-2-induced osteogenic differentiation in tendon-derived stem cells compared with bone-marrow-derived mesenchymal stem cells.

PURPOSE: Surgical reattachment of tendon to bone often fails due to regeneration failure of the specialised tendon-bone junction (TBJ). The use of mesenchymal stem cells for TBJ regeneration has been reported with promising results. Tendon-derived stem cells (TDSCs) with high proliferative and multi-lineage differentiation potential have been isolated. As stem cells residing in tendons, TDSCs can be considered a new cell source for TBJ repair. Bone morphogenic protein 2 (BMP-2) is a potent osteogenic factor with roles in normal bone healing and pathological ectopic bone formation in soft tissues. The use of BMP-2 to promote TBJ repair has been well reported. This study aimed to compare TDSCs to the gold standard bone-marrow-derived mesenchymal stem cells (BMSCs) with respect to osteogenic response to BMP-2 in vitro. METHOD: The clonogenicity and multi-differentiation potential of TDSCs and BMSCs were identified by colony-forming-unit assay, osteogenic, adipogenic and chondrogenic differentiation assays. Their osteogenic response to BMP-2 in vitro was examined by alkaline phosphatase (ALP) cytochemical staining, ALP activity assay and Alizarin red S staining of calcium nodule formation. Messenger RNA (mRNA) and BMP receptor (types IA, IB and II) protein expression were examined by quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. RESULTS: Our results showed that both TDSCs and BMSCs exhibited stem cell properties, including clonogenicity and multi-differentiation potential. TDSCs expressed higher mRNA and protein levels of BMP receptors IA, IB and II. They also exhibited higher osteogenic differentiation with and without BMP-2 stimulation compared with BMSCs. CONCLUSIONS: TDSCs with/without BMP-2 might be an attractive source for TBJ repair compared with BMSCs.

Upper limb muscle fatigue during prolonged Boccia games with underarm throwing technique.

This study investigated the acute fatigue pattern in neuromuscular activity after a simulated Boccia game and the effect of fatigue pattern on sport performance. Nine elite Boccia athletes were tested before, during, and after a simulated game. Maximum ball speed was captured with video, and the target hitting rate and rating of perceived exertion (RPE) score were collected and analyzed. Electromyography signals from the upper trapezius, anterior deltoid, posterior deltoid, biceps brachii, triceps brachii, and wrist extensor muscles were collected by surface electrode and were evaluated with mean power frequency (MPF). Only the upper trapezius muscle showed fatigue as demonstrated by a reduction of MPF of 8% (p = 0.027) when comparing the first and last throws in a simulated game. Subjective RPE score increased during the game (118%, p = 0.004), and sports performance in terms of maximum ball speed (-12%, p = 0.004) and target hitting rate (-25%, p = 0.004) also deteriorated. In conclusion, fatigue on the upper trapezius muscle was demonstrated in elite Boccia athletes following a prolonged Boccia game and may have affected Boccia performance. Preventative measures against upper trapezius muscle fatigue and endurance training for synergists of the upper trapezius muscle may be considered in future studies.

Comparison of 2 surgical techniques for reconstructing posterolateral corner of the knee: a cadaveric study evaluated by navigation system.

PURPOSE: This study aimed to evaluate the immediate effect on knee kinematics by 2 different techniques of posterolateral corner (PLC) reconstruction. METHODS: Five intact formalin-preserved cadaveric knees were used in this study. A navigation system was used to measure knee kinematics (posterior translation, varus angulation, and external rotation) after application of a constant force and torque to the tibia. Four different conditions of the knee were evaluated during the biomechanical test: intact knee and PLC-sectioned knee and PLC-reconstructed knee by the double-femoral tunnel technique and single-femoral tunnel technique. RESULTS: Sectioning of the PLC structures resulted in significant increases in external rotation at 30° of flexion from 11.2° (SD, 2.6) to 24.6° (SD, 6.2), posterior translation at 30° of flexion from 3.4 mm (SD, 1.5) to 7.4 mm (SD, 3.8), and varus angulation at 0° of flexion from 2.3° (SD, 2.1) to 7.9° (SD, 5.1). Both reconstruction techniques significantly restored the varus stability. The external rotation and posterior translation at 30° of flexion after reconstruction with the double-femoral tunnel technique were 10.2° (SD, 1.3) and 3.4° (SD, 2.7), respectively, which were significantly better than those of the single-femoral tunnel technique. CONCLUSIONS: Both techniques of reconstruction showed improved stability compared with PLC-sectioned knees. The double-femoral tunnel technique in PLC reconstruction showed better rotational stability and resistance to posterior translation than the single-femoral tunnel technique without compromising varus stability. CLINICAL RELEVANCE: PLC reconstruction by a double-femoral tunnel technique achieves better rotational control and resistance to posterior translation.

A rare mode of extensor mechanism failure in total knee arthroplasty.

We reported a rare mode of extensor mechanism failure in total knee arthroplasty. The patellar tendon was elongated and thin instead of disruption at the bone-tendon junction. We also described the surgical technique for reconstruction of patellar tendon. Patellar tendon was shortened by a precalculated amount. It was then augmented by autologous semitendinosus tendon graft and protected by tension band wire. Active full knee extension could be achieved at postoperative 10 months after the removal of wire loop.

rhBMP-2 not alendronate combined with HA-TCP biomaterial and distraction osteogenesis enhance bone formation.

INTRODUCTION: The long treatment duration of distraction osteogenesis (DO) usually causes some complications such as re-fracture, non-union. We have previously demonstrated that the combined use of biomaterial with distraction osteogenesis technique can enhance bone formation and consolidation. This study further tested whether the use of biological agents such as rhBMP-2 or alendronate together with biomaterials in DO will further promote bone formation. METHODS: A 1.0-cm tibial shaft was removed in the left tibia of 30 rabbits. The 1.0-cm defect gap was reduced to 0.5 cm and the remaining 0.5-cm defect gap was filled with 0.5-cm restorable hydroxyapatite/tri-calcium phosphates (HA/TCP) cylindrical block. The animals were divided into three groups with the following added on the HA/TCP block: Group A 50 µl of saline, Group B 75 µg rhBMP-2 in 50 µl of saline, Group C 250 µg alendronate in 50 µl saline. The tibia was then fixed with unilateral lengthener and lengthening started 7 days after at a rate of 1.0 mm/day for 5 days. All animals were terminated at day 37 following surgery. The excised bone specimens were subject to micro-CT, mechanical testing and histological examinations. RESULTS: Bone mineral density and content were significantly higher in Groups A and B compared to Group C and the mechanical properties of the regenerates in Group B were highest. Micro-CT and histological examinations also confirmed that the regenerates in Group B had the most advanced bone formation, consolidation and remodeling comparing to other groups. CONCLUSION: The combined use of rhBMP-2 with HA-TCP biomaterial in DO has significantly enhanced bone formation and consolidation than using the HA-TCP biomaterials alone, whereas the use of alendronate has inhibitory effects on bone formation.

Sustained expression of proteoglycans and collagen type III/type I ratio in a calcified tendinopathy model.

OBJECTIVES: Alteration in the composition of extracellular matrix has been suggested as the major factor for the development of tendinopathy and calcified tendinopathy, which has poorer clinical manifestation. This study investigated the changes of major proteoglycans and collagens in a calcified tendinopathy model and correlated the expression with the acquisition of chondrocyte phenotype, ectopic ossification and loss of matrix organization in the same model. METHODS: Thirty-six rats were used. Collagenase or saline was injected into the patellar tendons of each rat. At Weeks 2, 4 and 12, samples were used for immunohistochemistry of major proteoglycans and collagens and mRNA quantification. RESULTS: An increase in collagen type III and I expression was observed after injury at Week 2. Although their levels diminished with time, the ratio of collagen type III to collagen type I remained higher than that in healthy tendon at Week 12. The expression of biglycan, fibromodulin and aggrecan increased with time, whereas the expression of decorin was sustained from Week 2 to Week 12. The expression of major proteoglycans and collagens was observed in the tendon cells and matrix at Week 2 and became localized at the chondrocyte-like cells around the calcific deposits at Week 12. CONCLUSION: Sustained expression of proteoglycans and a high collagen type III/collagen type I ratio might account for poor matrix organization in calcified tendinopathy. The localization of major proteoglycans around chondro-osseous region might indicate interference of collagen assembly, which favours ectopic chondrogenesis, ossification and predisposition to tendon rupture.

Grafted tendon healing in tibial tunnel is inferior to healing in femoral tunnel after anterior cruciate ligament reconstruction: a histomorphometric study in rabbits.

PURPOSE: This study aimed to test whether graft healing in the tibial tunnel was inferior to that in the femoral tunnel after anterior cruciate ligament (ACL) reconstruction in rabbits. METHODS: Surgical reconstruction by use of the digital extensor tendon in the bone tunnel was performed in 18 rabbits. The rabbits were killed at weeks 2, 6, and 12 postoperatively, with 6 at each time point, for histologic examination. RESULTS: The transiently formed cartilaginous interface was gradually mineralized during re-establishment of direct tendon-to-bone integration, which was observed significantly less in the tibial tunnel than in the femoral tunnel (P < .05). The cell density of the graft was significantly lower in the tibial tunnel than that in the femoral tunnel at weeks 2 and 6 postoperatively (P < .05 for both). An increase in the immature type III collagen content was accompanied by a decrease in graft collagen fiber organization, with healing over time in both the femoral and tibial tunnels. The collagen fiber organization of the graft was significantly poorer in the tibial tunnel than that in the femoral tunnel at week 12 after surgery (P < .05). CONCLUSIONS: Grafted tendon healing in the tibial tunnel was inferior to that in the femoral tunnel at the tendon-to-bone interface and with regard to the grafted tendon within the bone tunnel after ACL reconstruction in rabbits. CLINICAL RELEVANCE: Future biopsy study is desirable to test whether this observation was valid clinically, which might provide a scientific basis for therapeutic targets to improve the outcome of ACL surgery.

In vivo low-intensity pulsed ultrasound (LIPUS) following tendon injury promotes repair during granulation but suppresses decorin and biglycan expression during remodeling.

STUDY DESIGN: Bench research, cross-sectional. OBJECTIVE: To determine if the effects of low-intensity pulsed ultrasound (LIPUS) on matrix synthesis change at different stages of tendon healing. BACKGROUND: LIPUS is effective in promoting tendon healing by stimulation of matrix synthesis. The timing of initiation and duration of LIPUS treatment have been shown to affect its effectiveness to promote tendon healing, suggesting a change of tissue responses to LIPUS stimulation. Understanding how the cellular responses to LIPUS stimulation change during tendon healing is thus important. METHODS: In a rat model of patellar tendon donor site injury, a single sonication of LIPUS or mock sonication was delivered to the injured knee of the rats on the fourth, 14th or 28th day postinjury. Tendon samples were harvested at 4 hours and 24 hours after sonication and the mRNA expression of COL1A1, COL3A1, decorin, biglycan, and TGF-beta 1 was analyzed. RESULTS: The results showed that a single sonication of LIPUS increased COL1A1 and COL3A1 mRNA in healing patellar tendons when administered on the fourth or 14th day postinjury, but not when administered on the 28th day postinjury. Both decorin and biglycan mRNA were decreased by treatment with LIPUS on the 28th day postinjury. Our results showed that LIPUS enhanced collagen synthesis in vivo only during the granulation phase. Matrix remodeling may be affected by LIPUS with the suppressed expression of decorin and biglycan. CONCLUSION: Our findings suggest that LIPUS should be applied during the granulation phase but not during the remodeling phase, to promote tendon healing.

Intra-Articular Delivery of Quercetin Using Thermosensitive Hydrogel Attenuate Cartilage Degradation in an Osteoarthritis Rat Model.

OBJECTIVE: Quercetin (Que), a bioflavonoid, is both anti-inflammatory and antioxidative. Que has been used as an oral supplement for osteoarthritis (OA) with inconsistent findings because of its low bioavailability. We encapsulated Que in a mPEG-polypeptide thermogel to prolong its bioactivity. The efficacy of this formulation was evaluated in a posttraumatic OA rat model. DESIGN: Methoxy-poly(ethylene glycol)-l-poly(alanine) (mPEG-PA) polymer was synthesized and characterized in terms of cytotoxicity and release kinetics in vitro. At 12 weeks old, Sprague-Dawley rats underwent anterior cruciate ligament transection (ACLT). At 24 weeks post-operation, rats received either an intra-articular (IA) injection of saline, hydrogel, or hydrogel with Que (50 or 500 µg). Gait analysis was performed at pre-ACLT, pre-treatment, and at 4, 8, and 12 weeks post-treatment. At 12 weeks post-treatment, knee joints were collected for histopathological evaluation. RESULTS: In vitro studies showed that chondrocytes were viable after 72 hours of incubation with mPEG-PA, and the release of Que could be sustained for >28 days. Among all OA rats, the limb idleness index (LII) were significantly increased at 24 weeks post-ACLT. Rats that received hydrogel with Que (50 µg) showed the most reduction in LII at both 4 and 8 weeks post-treatment. The Osteoarthritis Research Society International score of rats received hydrogel with Que (50 µg) was significantly lower than the control group. All rats suffered from low-grade synovitis (Krenn score: 2-4). CONCLUSION: This study suggests that a sustained delivery of Que (50 µg) could provide symptom relief and also delay the progression of OA in the knee.