Snapshot of degenerative aging of porcine intervertebral disc: a model to unravel the molecular mechanisms

조홍식,김송자,박상혁,Sangmin Lee,Miji Kang,Karen A. Hasty 생화학분자생물학회 2011 Experimental and molecular medicine Vol.43 No.6

Larger animal models, such as porcine, have been validated as appropriate models of the human disc with respect to biomechanics and biochemistry. They are advantageous for research as the models are relatively straightforward to prepare and easily obtainable for research to perform surgical techniques. The intention of this study was to quantitatively analyze gene expression for collagen and proteoglycan components of the extracellular matrix and for collagenase (MMP-1)in porcine discs of varying ages (Newborn; 2-3weeks,Mature; 6-9 month, Older; 2-3 years). In this study, we observed that the cell number and GAG (glycosaminoglycan)formation dramatically decreased with aging. Also, gene expression in the annulus fibrosus (AF) and nucleus pulposus (NP) cells changed with aging. The level of MMP-1 mRNA increased with age and both type I, II collagens decreased with age. The level of aggrecan mRNA was highest in the mature group and decreased significantly with aging. In the mature group, MMP-1expression was minimal compared to the newborn group. In AF cells, type II collagen was expressed at a high level in the mature group with a higher level of aggrecan,when aged NP showed a decrease in type II collagen. The model of IVD degeneration in the porcine disc shows many changes in gene expression with age that have been previously documented for human and may serve as a model for studying changes in IVD metabolism with age. We concluded that the porcine model is excellent to test hypotheses related to disc degeneration while permitting time-course study in biologically active systems.

Immobilization of Fibrinogen Antibody on Self-Assembled Gold Monolayers for Immunosensor Applications

조홍식,Justin Zook,Todd Banner,박상혁,민병현,Karen A. Hasty,Eugene Pinkhassik,Erno Lindner 한국조직공학과 재생의학회 2014 조직공학과 재생의학 Vol.11 No.1

Self-assembled gold monolayers offer several advantages for the realization of novel modified electrodes for biosensor applications. This is due to their ability to decrease non-specific adsorption and provide for covalent attachment of biomolecules. Surfaces for these applications require the precise control of ligand density, the ability to immobilize ligands, and in situ-modulation of ligand activity. In this study, we focused our studies on the immobilization of antibody on a gold monolayer surface. We self-assembled thioctic acid onto the gold surface as an anchor point for the immobilization of anti-fibrinogen onto the surface. The modifications to the gold surface were characterized by ELISA, ellipsometry, and AFM.

Cytoprotective role of vitamin E in porcine adipose-tissue-derived mesenchymal stem cells against hydrogen-peroxide-induced oxidative stress

Bhatti, Fazal Ur Rehman,Kim, Song Ja,Yi, Ae-Kyung,Hasty, Karen A.,Cho, Hongsik Springer-Verlag 2018 Cell and tissue research Vol.374 No.1

Synergistic Effect of Combined Growth Factors in Porcine Intervertebral Disc Degeneration

Cho, Hongsik,Lee, Sangmin,Park, Sang-Hyug,Huang, Jinsong,Hasty, Karen A.,Kim, Song-Ja Informa Healthcare 2013 Connective tissue research Vol.54 No.3

<P>Although intervertebral disc (IVD) degeneration is one of most common causes of morbidity, its etiology remains unclear. In healthy discs, the rates of synthesis and breakdown of the extracelluar matrix (ECM) are in equilibrium because of intricate regulation by growth factors and catabolic cytokines. Important among these physiologic growth factors are transforming growth factor-β (TGF-β1) and bone morphogenetic protein-2 (BMP-2). Disc degeneration is thought to be associated with a loss of this homeostasis between proteoglycan (PG) synthesis and cytokine-induced degradation leading to up-regulation of matrix metalloproteinases (MMP) families and down-regulation of extracelluar matrix production. Several strategies using biological agents have been attempted to manage IVD degeneration, improving the function and anabolic capabilities of IVD cells and inhibiting matrix degradation. The purpose of this study is to compare the effects of the anabolic cytokines BMP-2 and TGF-β1 with those of the catabolic cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) on porcine annulus fibrosus (AF). The results of this study show that the application of pro-inflammatory cytokines like tumor necrosis factor-α and interleukin-1β to normal annulus fibrosus cells leads to a significant increase in tissue levels of the degradative protease MMP-1. Treatment with a combination of minimum doses of both BMP-2 and TGF-β1 caused a greater decrease in MMP-1 and increase in aggrecan than either cytokine alone, suggesting a synergistic effect of the combined cytokines.</P>

Noninvasive visualization of early osteoarthritic cartilage using targeted nanosomes in a destabilization of the medial meniscus mouse model

Cho, Hongsik,Kim, Byoung Ju,Park, Sang-Hyug,Hasty, Karen A,Min, Byoung-Hyun Dove Medical Press 2018 INTERNATIONAL JOURNAL OF NANOMEDICINE Vol.13 No.-

<P><B>Background</B></P><P>Early stage osteoarthritis (OA) is clinically asymptomatic due to the avascular and the aneural nature of the cartilage tissue. Nevertheless, early detection of cartilage tissue is critical in order to impede the progression of OA. Hence, in order to develop effective preventive therapy for OA, diagnosis in the early stages is necessary.</P><P><B>Methods</B></P><P>To achieve this goal, we have developed targeted, fluorescent nanosomes conjugated with monoclonal anti-type II collagen antibodies (MabCII) for diagnosis of early OA. The MabCII-coated nanosomes (targeted-nanosomes) bind to the damaged cartilage explants in vitro and in vivo in an OA mouse model that mimics early stage OA. The OA mouse model was induced by destabilization of the medial meniscus (DMM) in 9–10 weeks old C57Bl/6 mice.</P><P><B>Results</B></P><P>The targeted-nanosomes enhanced the binding specificity to the cartilage tissue according to the severity of damage.</P><P><B>Conclusion</B></P><P>We show that MabCII-nanosomes can precisely detect early stage OA in the DMM mouse model. Thus, MabCII-nanosomes have the potential to be used as a non-invasive method for diagnosing the early osteoarthritic lesions.</P>

Snapshot of degenerative aging of porcine intervertebral disc: a model to unravel the molecular mechanisms

Cho, Hong-Sik,Park, Sang-Hyug,Lee, Sang-Min,Kang, Mi-Ji,Hasty, Karen A.,Kim, Song-Ja Korean Society for Biochemistry and Molecular Bion 2011 Experimental and molecular medicine Vol.43 No.6

Larger animal models, such as porcine, have been validated as appropriate models of the human disc with respect to biomechanics and biochemistry. They are advantageous for research as the models are relatively straightforward to prepare and easily obtainable for research to perform surgical techniques. The intention of this study was to quantitatively analyze gene expression for collagen and proteoglycan components of the extracellular matrix and for collagenase (MMP-1) in porcine discs of varying ages (Newborn; 2-3weeks, Mature; 6-9 month, Older; 2-3 years). In this study, we observed that the cell number and GAG (glycosaminoglycan) formation dramatically decreased with aging. Also, gene expression in the annulus fibrosus (AF) and nucleus pulposus (NP) cells changed with aging. The level of MMP-1 mRNA increased with age and both type I, II collagens decreased with age. The level of aggrecan mRNA was highest in the mature group and decreased significantly with aging. In the mature group, MMP-1 expression was minimal compared to the newborn group. In AF cells, type II collagen was expressed at a high level in the mature group with a higher level of aggrecan, when aged NP showed a decrease in type II collagen. The model of IVD degeneration in the porcine disc shows many changes in gene expression with age that have been previously documented for human and may serve as a model for studying changes in IVD metabolism with age. We concluded that the porcine model is excellent to test hypotheses related to disc degeneration while permitting time-course study in biologically active systems.

Scaffold-Free, Engineered Porcine Cartilage Construct for Cartilage Defect Repair—In Vitro and In Vivo Study

Park, Kwideok,Huang, Jinsong,Azar, Frederick,Jin, Ri L.,Min, Byoung-Hyun,Han, Dong K.,Hasty, Karen Blackwell Publishing Inc 2006 Artificial Organs Vol.30 No.8

<P>Abstract: </P><P>This study introduces an implantable scaffold-free (SF) cartilage tissue construct that is composed of chondrocytes and their self-produced extracellular matrix (ECM). Chondrocytes were isolated from the articular cartilages from knees of domestic pigs (2-week old) and monolayer-cultured for 3–4 days in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal calf serum and 50 µg/mL of ascorbic acid. Briefly treated with 0.25% trypsin–ethylenediaminetetraacetic acid (EDTA), an intact chondrocytes/ECM membrane, as a cell sheet was released from the plate bottom and subsequently centrifuged into a pellet-type construct. Each was grown in vitro for up to 5 weeks and subjected to various assays at different time points (1, 7, 14, 21, and 35 days). For in vivo implantation, full-thickness defects (<I>n</I> = 4) were manually created on the femoro-patellar groove of the left porcine knee and 1-week-cultured SF construct was implanted as an allograft for a month. One defect (♯1) was an empty control and the remaining three received different recipes; construct only (♯2) or 0.25% trypsin/EDTA-treated first and then construct and collagen gel (♯3) or construct and collagen gel (♯4). While the total cell numbers significantly increased by 2 weeks and then remained stable, cell viability stayed in the mid-70% range through the entire culture period. Biochemical assay found continuous glycosaminoglycan (GAG) accumulation. Histology exhibited that cell distribution was even in the construct and GAG intensity became stronger and uniform with time. Real-time reverse transcription polymerase chain reaction (RT-PCR) results showed that phenotypic stability peaked at 2 weeks, which was arable to that of freshly isolated chondrocytes. Upon analysis of the retrieved implants, some promising results were witnessed in the defects (♯3) retaining not only their intact mass but also chondrocytic morphology with lacuna formation. </P>

Variation of sexual dimorphism and asymmetry in disease expression of inflammatory arthritis among laboratory mouse models with different genomic backgrounds

Wei Dong,Cheng Tian,Z. Galvin Li,David Brand,Yanhong Cao,Xiaoyun Liu,Jiamin Ma,Andy Chai,LindaK.myers,Jian Yan,Karen Hasty,John Stuart,Yan Jiao,Weikuan Gu,Xiaojun Cai 한국실험동물학회 2023 Laboratory Animal Research Vol.39 No.4

Sex difference has shown in the arthritis diseases in human population and animal models. We investigate how the sex and symmetry vary among mouse models with different genomic backgrounds. Disease data of sex and limbs accumulated in the past more than two decades from four unique populations of murine arthritis models were analyzed. They are (1) interleukin-1 receptor antagonist (IL-1ra) deficient mice under Balb/c background (Balb/c KO); (2) Mice with collagen II induced arthritis under DBA/1 background; (3) Mice with collagen II induced arthritis under C57BL/6 (B6) background and (4) A F2 generation population created by Balb/c KO X DBA/1 KO. Our data shows that there is a great variation in sexual dimorphism for arthritis incidence and severity of arthritis in mice harboring specific genetic modifications. For a F2 population, the incidence of arthritis was 57.1% in female mice and 75.6% in male mice. There was a difference in severity related to sex in two populations: B6.DR1/ B6.DR4 (P < 0.001) and F2 (P = 0.023) There was no difference Balb/c parental strain or in collagen-induced arthritis (CIA) in DBA/1 mice. Among these populations, the right hindlimbs are significantly higher than the scores for the left hindlimbs in males (P < 0.05). However, when examining disease expression using the collagen induced arthritis model with DBA/1 mice, sex-dimorphism did not reach statistical significance, while left hindlimbs showed a tendency toward greater disease expression over the right. Sexual dimorphism in disease expression in mouse models is strain and genomic background dependent. It sets an alarm that potential variation in sexual dimorphism among different racial and ethnic groups in human populations may exist. It is important to not only include both sexes and but also pay attention to possible variations caused by disease expression and response to treatment in all the studies of arthritis in animal models and human populations.