scholarly journals Comparative Transcriptomics Identifies Novel Genes and Pathways Involved in Post-Traumatic Osteoarthritis Development and Progression

2018 ◽  
Author(s):  
Aimy Sebastian ◽  
Jiun C. Chang ◽  
Melanie E. Mendez ◽  
Deepa K. Murugesh ◽  
Sarah Hatsell ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Mechanisms ◽  
Cruciate Ligament ◽  
Cartilage Damage ◽  
Acl Injury ◽  
The Other ◽  
Knee Joints ◽  
Mouse Strains ◽  
Post Injury ◽  
Post Traumatic

Injuries to the anterior cruciate ligament (ACL) often result in post-traumatic osteoarthritis (PTOA). To better understand the molecular mechanisms behind PTOA development following ACL injury, we profiled ACL injury-induced gene expression changes in knee joints of three mouse strains with varying susceptibility to OA: STR/ort (highly susceptible), C57BL/6 (moderately susceptible) and super-healer MRL/MpJ (not susceptible). Right knee joints of the mice were injured using a non-invasive tibial compression injury model that closely mimics ACL rupture in humans and global gene expression was quantified before and at 1-day, 1-week, and 2-weeks post-injury using RNA-seq. Following injury, STR/ort displayed severe cartilage degeneration while MRL/MpJ had little cartilage damage. Gene expression analysis suggested that prolonged inflammation and elevated catabolic activity in STR/ort injured joints, compared to the other two strains may be responsible for the severe PTOA phenotype observed in this strain. MRL/MpJ had the lowest expression values for several inflammatory cytokines and catabolic enzymes activated in response to ACL injury. Furthermore, we identified several genes highly expressed in MRL/MpJ compared to the other two strains including B4galnt2 and Tpsab1 which may contribute to enhanced healing in the MRL/MpJ. Overall, this study has increased our knowledge of early molecular changes associated with PTOA development.

scholarly journals Comparative Transcriptomics Identifies Novel Genes and Pathways Involved in Post-Traumatic Osteoarthritis Development and Progression

2018 ◽  
Vol 19 (9) ◽  
pp. 2657 ◽  
Author(s):  
Aimy Sebastian ◽  
Jiun Chang ◽  
Melanie Mendez ◽  
Deepa Murugesh ◽  
Sarah Hatsell ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Mechanisms ◽  
Cruciate Ligament ◽  
Cartilage Damage ◽  
Acl Injury ◽  
The Other ◽  
Knee Joints ◽  
Mouse Strains ◽  
Post Injury ◽  
Post Traumatic

Anterior cruciate ligament (ACL) injuries often result in post-traumatic osteoarthritis (PTOA). To better understand the molecular mechanisms behind PTOA development following ACL injury, we profiled ACL injury-induced transcriptional changes in knee joints of three mouse strains with varying susceptibility to OA: STR/ort (highly susceptible), C57BL/6J (moderately susceptible) and super-healer MRL/MpJ (not susceptible). Right knee joints of the mice were injured using a non-invasive tibial compression injury model and global gene expression was quantified before and at 1-day, 1-week, and 2-weeks post-injury using RNA-seq. Following injury, injured and uninjured joints of STR/ort and injured C57BL/6J joints displayed significant cartilage degeneration while MRL/MpJ had little cartilage damage. Gene expression analysis suggested that prolonged inflammation and elevated catabolic activity in STR/ort injured joints, compared to the other two strains may be responsible for the severe PTOA phenotype observed in this strain. MRL/MpJ had the lowest expression values for several inflammatory cytokines and catabolic enzymes activated in response to ACL injury. Furthermore, we identified several genes highly expressed in MRL/MpJ compared to the other two strains including B4galnt2 and Tpsab1 which may contribute to enhanced healing in the MRL/MpJ. Overall, this study has increased our knowledge of early molecular changes associated with PTOA development.

scholarly journals Effect of knee joint loading on chondrocyte mechano-vulnerability and severity of post-traumatic osteoarthritis induced by ACL-injury in mice

2021 ◽  
Author(s):  
Alexander Kotelsky ◽  
Anissa Elahi ◽  
Nejat Can ◽  
Ashley Proctor ◽  
Sandeep Mannava ◽  
...  
Keyword(s):  
Articular Chondrocytes ◽  
Cruciate Ligament ◽  
Weight Bearing ◽  
Acl Injury ◽  
Joint Loading ◽  
Post Injury ◽  
Chondrocyte Death ◽  
Hind Limbs ◽  
Post Traumatic

Objective: The objective of this study is to understand the role of altered in vivo mechanical environments in knee joints post anterior cruciate ligament (ACL)-injury in chondrocyte vulnerability against mechanical stimuli and in the progression of post-traumatic osteoarthritis (PT-OA). Methods: Differential in vivo mechanical environments were induced by unilateral ACL-injury (uni-ACL-I) and bilateral ACL-injury (bi-ACL-I) in 8-week-old female C57BL/6 mice. The gait parameters, the mechano-vulnerability of in situ chondrocytes, Youngs moduli of cartilage extracellular matrix (ECM), and the histological assessment of OA severity (OARSI score) were compared between control and experimental groups at 0~8-weeks post-ACL-injury. Results: We found that bi-ACL-I mice experience higher joint-loading on their both injured limbs, but uni-ACL-I mice balance their joint-loading between injured and uninjured hind limbs resulting in a reduced joint-loading during gait. We also found that at 4- and 8-week post-injury the higher weight-bearing hind limbs (i.e., bi-ACL-I) had the increased area of chondrocyte death induced by impact loading and higher OARSI score than the lower weight-bearing limbs (uni-ACL-I). Additionally, we found that at 8-weeks post-injury the ECM became stiffer in bi-ACL-I joints and softer in uni-ACL-I joints. Conclusions: Our results show that ACL-injured limbs with lower in vivo joint-loading develops PT-OA significantly slower than injured limbs with higher joint-loading during gait. Our data also indicate that articular chondrocytes in severe PT-OA are more fragile from mechanical impacts than chondrocytes in healthy or mild PT-OA. Thus, preserving physiologic joint-loads on injured joints will reduce chondrocyte death post-injury and may delay PT-OA progression.

R432 – Gene Expression of the Remodeling Rat Vocal Fold Wound

2008 ◽  
Vol 139 (2_suppl) ◽  
pp. P189-P189
Author(s):  
Tsunehisa Ohno ◽  
Lesley C. French ◽  
Bernard Rousseau
Keyword(s):  
Gene Expression ◽  
Extracellular Matrix ◽  
Messenger Rna ◽  
Vocal Fold ◽  
Molecular Mechanisms ◽  
Type I ◽  
Post Injury ◽  
Time Points ◽  
Procollagen Type ◽  
Procollagen Type I

Problem The authors investigated the expression of key extracellular matrix genes after vocal fold wounding in a rat model to better understand the reparative mechanisms of tissue repair during the remodeling phase of vocal fold injury. Methods Bilateral vocal fold wounds were created in 30 rats. Injured vocal fold specimens were harvested 1, 3, 7, 14, 28, and 56 days after wounding. 5 unwounded rats were used to establish baseline for polymerase chain reaction (PCR). The authors used real-time PCR to quantify messenger RNA expression of procollagen type I, III, interleukin-1 beta (IL-1 beta), decorin, and hyaluronan synthase (HAS) −1, −2, and −3. Analysis of variance was used to detect main effects for gene expression. Post-hoc tests were used to make comparisons between time points. Results Procollagen type I expression was decreased from baseline on post-injury day 1, 28, and 56. Procollagen type III was decreased on post-injury day 1 and 56, and increased from baseline on post-injury day 14. IL-1 beta expression was increased from baseline on post-injury day 1, 3, and 7. Decorin expression was decreased from baseline on post-injury day 1, 3, 7, and 56. HAS-1 expression was decreased from baseline at all post-injury time points. HAS-2 expression was increased from baseline on post-injury day 3, and decreased from baseline on post-injury day 14, 28, and 56. HAS-3 expression was decreased from baseline on post-injury day 1, 28, and 56. Conclusion Findings provide temporal changes in the expression of key extracellular matrix genes during a remodeling phase of vocal fold injury in a rat wound model. Significance Vocal fold wound models provide a means for investigating tissue reparative processes and molecular mechanisms controlling synthesis and degradation of the vocal fold extracellular matrix. Support Vanderbilt University Medical Center.

scholarly journals One-leg rise performance and associated knee kinematics in ACL-deficient and ACL-reconstructed persons 23 years post-injury

2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Andrew Strong ◽  
Eva Tengman ◽  
Divya Srinivasan ◽  
Charlotte K. Häger
Keyword(s):  
Cruciate Ligament ◽  
Scientific Evidence ◽  
Knee Kinematics ◽  
Acl Injury ◽  
Cross Sectional Study ◽  
Knee Function ◽  
Post Injury ◽  
Discriminative Ability ◽  
Cross Sectional ◽  
Knee Abduction

Abstract Background Research indicates reduced knee function and stability decades after anterior cruciate ligament (ACL) injury. Assessment requires reliable functional tests that discriminate such outcomes from asymptomatic knees, while providing suitable loading for different populations. The One-leg rise (OLR) test is common in clinics and research but lacks scientific evidence for its implementation. Our cross-sectional study compared performance including knee kinematics of the OLR between ACL-injured persons in the very long term to controls and between legs within these groups, and assessed the within-session reliability of the kinematics. Methods Seventy ACL-injured individuals (mean age 46.9 ± 5.4 years) treated with either reconstructive surgery and physiotherapy (ACLR; n = 33) or physiotherapy alone (ACLPT; n = 37), on average 23 years post-injury, and 33 age- and sex-matched controls (CTRL) attempted the OLR. Participants completed as many repetitions as possible to a maximum of 50 while recorded by motion capture. We compared between all groups and between legs within groups for total repetitions and decomposed the OLR into movement phases to compare phase completion times, maximum and range of knee abduction and adduction angles, and mediolateral knee control in up to 10 repetitions per participant. Results ACLPT performed significantly fewer OLR repetitions with their injured leg compared to the CTRL non-dominant leg (medians 15 and 32, respectively) and showed significantly greater knee abduction than ACLR and CTRL (average 2.56°-3.69° depending on phase and leg). Distribution of repetitions differed between groups, revealing 59% of ACLPT unable to complete more than 20 repetitions on their injured leg compared to 33% ACLR and 36% CTRL for their injured and non-dominant leg, respectively. Within-session reliability of all kinematic variables for all groups and legs was high (ICC 3,10 0.97–1.00, 95% CI 0.95–1.00, SEM 0.93–1.95°). Conclusions Negative outcomes of OLR performance, particularly among ACLPT, confirm the need to address aberrant knee function and stability even decades post-ACL injury. Knee kinematics derived from the OLR were reliable for asymptomatic and ACL-injured knees. Development of the OLR protocol and analysis methods may improve its discriminative ability in identifying reduced knee function and stability among a range of clinical populations.

scholarly journals Quantitative assessment of neural elements in a rat model using nerve growth factor after remnant-preserving anterior cruciate ligament reconstruction:a haematoxylin and eosin staining and immunofluorescence study

2020 ◽  
Author(s):  
Sung Hyun Lee ◽  
Hyung Gyu Cho ◽  
Jin Soo Song ◽  
Keun Churl Chun ◽  
Churl Hong Chun
Keyword(s):  
Anterior Cruciate Ligament ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Achilles Tendon ◽  
Acl Reconstruction ◽  
Cruciate Ligament ◽  
The Other ◽  
Knee Joints ◽  
Neural Cells ◽  
Anterior Cruciate

Abstract Background: Immunofluorescence analyses of anterior cruciate ligament (ACL) allografts following remnant-preserving ACL reconstruction using Achilles tendon allografts have provided evidence for the presence of neural elements. In this study, we aimed to examine the expression of neural elements and quantify the presence of neural cells in ACL remnants and Achilles allografts using nerve growth factor (NGF) therapy after remnant-preserving ACL reconstruction.Methods: Experiments were conducted on 5 pairs of rats (approximately 8 weeks old and weighing 320 g at the time of surgery). Longitudinally split Achilles tendons from the paired rats were freshly frozen and later defrosted with warm saline and allografted onto the right ACL of the other rat that was partially detached at the femoral attachment site. A sham operation was conducted on the left knee to be used as a control. NGF was injected into both knee joints every week for 6 weeks after surgery. The presence of neural cells in the ACL of the sham-operated knee, allografted Achilles tendon, and ACL remnant was examined 6 weeks post-surgery using H and E and immunofluorescent staining.Results: H and E staining did not reveal neural cells in any of the three groups. However, immunofluorescence analysis showed the presence of nestin-positive neural elements in the normal ACL tissues as well as ACL remnants. Additionally, neural elements were examined in 7 of the 8 (87.5%) allograft tissues. Quantitative analysis showed no difference in the number or area of nuclei among the three groups. However, the number and area of neural cells in the Achilles allografts were significantly lower than those in the other two groups (p=0.000 and p=0.001, respectively).Conclusion: Our observations indicate that ACL remnants promote the new ingrowth and persistence of neural cells. We suggest that the ingrowth of neural elements can support the persistence and new ingrowth of mechanoreceptors, thereby enhancing the functional stability of knee joints. Moreover, the expression of neural cells in the Achilles allografts was lower than that in normal ACL tissues and ACL remnants in the quantitative evaluation, thereby confirming the essential role of ACL remnants in knee joint functionalization.

scholarly journals The Incidence of Post-Traumatic Osteoarthritis on Clinical Radiographs at 10 Years after Anterior Cruciate Ligament Reconstruction: Data from the MOON Nested Cohort (168)

2021 ◽  
Vol 9 (10_suppl5) ◽  
pp. 2325967121S0029
Author(s):  
Joshua Everhart ◽  
Morgan Jones ◽  
Sercan Yalcin ◽  
Emily Reinke ◽  
Laura Huston ◽  
...  
Keyword(s):  
Ligament Reconstruction ◽  
Cruciate Ligament ◽  
Acl Injury ◽  
Average Difference ◽  
Cruciate Ligament Reconstruction ◽  
Osteophyte Formation ◽  
Post Traumatic ◽  
Anterior Cruciate ◽  
Intact Knee

Objectives: 1) To prospectively determine the incidence of post-traumatic osteoarthritis (PTOA) at 10 years after anterior cruciate ligament reconstruction (ACLR) in young athletic patients on clinical radiographs: and 2) to determine the average difference in clinical radiographic osteoarthritis changes (joint space narrowing [JSN] and osteophyte formation) between the ACLR and contralateral ACL-intact knees. Methods: The first 146 patients in an ongoing prospective nested cohort study within the Multicenter Orthopaedic Outcomes Network (MOON) cohort returned onsite for minimum 10-year follow-up. Inclusion criteria were that patients had a sports-related ACL injury, no prior history of knee surgery, no contralateral ACL injury, and were less than 33 years of age at the time of their ACLR. Bilateral knee standing metatarsophalangeal (MTP) view radiographs were obtained and graded by International Knee Documentation Committee (IKDC), Osteoarthritis Research Society International (OARSI), and modified Kellgren-Lawrence (KL) criteria by two blinded reviewers. Inter-rater reliability was determined for all clinical radiographic OA grading criteria. The incidence and severity of ipsilateral and contralateral knee osteoarthritis were determined among patients without a contralateral ACL injury before 10-year follow-up (n=133). Results: Inter-rater reliability was substantial for IKDC (Gwet’s AC1 = 0.71), moderate for KL (0.48) and almost perfect for OARSI (0.84) grading systems. The 10-year incidence of PTOA on clinical radiographs in the ACLR knee was 43% as defined by osteophytes and 27% as defined by JSN (Table 1). In the contralateral ACL-intact knee, the incidence of osteophyte-defined OA was 10% and JSN-defined OA was 5%. The maximum side-to side difference in medial or lateral compartment OARSI osteophyte grade was 0 in 65% of patients, 1 in 20%, and 2+ in 15% (Figure 1) (Table 2). The maximum difference in OARSI JSN grade was 0 in 77% of patients, 1 in 19%, and 2+ in 4% (Figure 2) (Table 2). Conclusions: In young active patients, the 10-year incidence on clinical radiographs of osteophyte-defined PTOA after ACLR is 43% and JSN-defined PTOA is 27%. The average difference in degree of osteophyte formation (≤1 grade in 85%) and JSN (≤1 grade in 96%) between the ACLR knee and contralateral ACL-intact knee is small.

scholarly journals Global Gene Expression Analysis Identifies Age-Related Differences in Knee Joint Transcriptome during the Development of Post-Traumatic Osteoarthritis in Mice

2020 ◽  
Vol 21 (1) ◽  
pp. 364 ◽  
Author(s):  
Aimy Sebastian ◽  
Deepa K. Murugesh ◽  
Melanie E. Mendez ◽  
Nicholas R. Hum ◽  
Naiomy D. Rios-Arce ◽  
...  
Keyword(s):  
Knee Joint ◽  
Age Groups ◽  
Knee Joints ◽  
Compression Injury ◽  
Post Injury ◽  
Transcriptional Responses ◽  
Molecular Changes ◽  
Age Related ◽  
Post Traumatic ◽  
Old Mice

Aging and injury are two major risk factors for osteoarthritis (OA). Yet, very little is known about how aging and injury interact and contribute to OA pathogenesis. In the present study, we examined age- and injury-related molecular changes in mouse knee joints that could contribute to OA. Using RNA-seq, first we profiled the knee joint transcriptome of 10-week-old, 62-week-old, and 95-week-old mice and found that the expression of several inflammatory-response related genes increased as a result of aging, whereas the expression of several genes involved in cartilage metabolism decreased with age. To determine how aging impacts post-traumatic arthritis (PTOA) development, the right knee joints of 10-week-old and 62-week-old mice were injured using a non-invasive tibial compression injury model and injury-induced structural and molecular changes were assessed. At six-week post-injury, 62-week-old mice displayed significantly more cartilage degeneration and osteophyte formation compared with young mice. Although both age groups elicited similar transcriptional responses to injury, 62-week-old mice had higher activation of inflammatory cytokines than 10-week-old mice, whereas cartilage/bone metabolism genes had higher expression in 10-week-old mice, suggesting that the differential expression of these genes might contribute to the differences in PTOA severity observed between these age groups.

Identification of New Megakaryocytic Subpopulations in Mouse Bone Marrow.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 2265-2265
Author(s):  
Shinji Sogo ◽  
Kuniko Matsumura-Takeda ◽  
Yoshimasa Isakari ◽  
Yasuo Harada ◽  
Kinue Nishioka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Gene Expression Profiling ◽  
Expression Profiling ◽  
Molecular Mechanisms ◽  
Present Finding ◽  
The Other ◽  
Mouse Bone Marrow ◽  
Rt Pcr ◽  
Kit Gene

Abstract Platelets (PLT) are produced from megakaryocytes (Mks) via proplatelet formation (PPF). However, the molecular mechanisms from Mks to PPF are not clearly elucidated, because the maturational steps of the Mks in bone marrow (BM) are not analyzed in detail. Until now, mouse Mks have been only isolated as acetylcholinesterase (AchE) positive cells and they are understood as well maturated population. In this study, we found the presence of different megakaryocytic subpopulations in BM by flowcytometry. To isolate the Mks, first we depleted lineage marker (CD4, CD8a, CD11b, B220, CD71, CD90, TER119, Gr-1, F4/80, 7/4) positive cells from BM cells of BALB/c mice. The analysis of the expression-pattern of CD41, CD45 and CD61 in the lineage negative (Lin−) cells showed the presence of two types of megakaryocytic subpopulations. By sorting, they were identified as Lin−CD41+/45+/61+ cells (AchE negative) and Lin−CD41++/45+/61++ cells (partially AchE positive), respectively. To assess the maturational stages of the subpopulations, each population was cultured with 10ng/mL of TPO followed by counting of PPF and PLT production. Both PPF and PLT production were observed in Lin−CD41+/45+/61+ cells later than those in Lin−CD41++/45+/61++ cells. On the other hand, CFU-Mk was scarcely detected in each subpopulation. The results indicate that both populations are the committed megakaryocytes and Lin−CD41+/45+/61+ cells are more immature population than Lin−CD41++/45+/61++ cells. Then to characterize these subpopulations in detail, gene expression profiling was performed against four-megakaryocytic lineage-populations, Lin−CD41−Thy1lowc-kit+ cells as stem/progenitor, Lin−CD41+/45+/61+ cells, Lin−CD41++/45+/61++ cells and PLT using GeneChipU74 or RT-PCR. These analyses revealed that many PLT-specific genes including gpIb/IX, P-selectin, thrombin-R and ADP-R were already expressed on Lin−CD41+/45+/61+ cells but less than Lin−CD41++/45+/61++ cells. Especially, beta-1 tubulin that is necessary for PPF was only expressed on Lin−CD41++/45+/61++ cells. On the contrary, the expression of c-kit gene was gradually decreasing from stem/progenitor fraction to PLT. In conclusion, we succeeded in the isolation of new subpopulations distinguishable between immature Mks and more matured Mks beginning to prepare PLT. The present finding can contribute to elucidate the molecular mechanisms during terminal maturation.

scholarly journals Quantitative assessment of neural elements in rat model using nerve growth factor after remnant-preserving anterior cruciate ligament reconstruction: a hematoxylin and eosin stain and immunohistochemical study

2020 ◽  
Author(s):  
Sung Hyun Lee ◽  
Hyung Gyu Cho ◽  
Jin Soo Song ◽  
Keun Churl Chun ◽  
Churl Hong Chun
Keyword(s):  
Anterior Cruciate Ligament ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Achilles Tendon ◽  
Cruciate Ligament ◽  
The Other ◽  
Knee Joints ◽  
Neural Cells ◽  
Nerve Growth ◽  
Anterior Cruciate

Abstract Background: Immunohistochemical analyses of anterior cruciate ligament (ACL) allografts following remnant-preserving ACL reconstructions using Achilles tendon allografts have provided evidence for the presence of neural elements. In this study, we aimed to examine the expression of neural elements and quantify the presence of neural cells in ACL remnants and Achilles allografts using nerve growth factor (NGF) therapy after remnant-preserving ACL reconstruction. Methods: Experiments were conducted on 5 pairs of rats (approximately 8 weeks old and weighting 320 g at the time of surgery). Longitudinally-split Achilles tendons from the paired rats were freshly frozen and later defrosted with warm saline and allografted onto the right ACL of the other, which was partially detached at the femoral attachment site. A sham operation was conducted on the left knee to be used as Control. NGF was injected in both the knee joints 1 week after surgery. The presence of neural cells in the ACL of the sham-operated knee, allografted Achilles tendon, and ACL remnant was examined 6 weeks post surgery using H and E and immunohistochemical staining. Results: H and E staining did not reveal neural cells in any of the three groups. However, immunohistochemical analysis showed the presence of nestin-positive neural elements in normal ACL as well as ACL remnants. Additionally, neural elements were examined in 7 of the 8 (87.5%) allograft tissues. Quantitative analysis showed no difference in the number and area of nuclei among the three groups. However, the number and area of neural cells in Achilles allograft were significantly lower than in the other two groups (p=0.000 and p=0.001, respectively). Conclusion: Our observations indicate that ACL remnants promote new ingrowth and persistence of neural cells. We suggest that the ingrowth of neural elements could support the persistence and new ingrowth of mechanoreceptors, thereby enhancing the functional stability of knee joints. Moreover, the expression of neural cells in Achilles allograft was lower than that of normal ACL or ACL remnants in the quantitative evaluation, thereby confirming the essential role of ACL remnants in knee joint functionalization. Key terms: anterior cruciate ligament, remnant preservation, immunohistochemistry, nerve growth factor

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