Preconditioning of Rabbit Mesenchymal Stem Cells in Polyglycolic Acid (PGA) Scaffold using Low-Intensity Ultrasound Improved Regeneration of Cartilage in Rabbit Articular Cartilage Defect Model

( Ji Hao Cui ),( So Ra Park ),( Byung Hyune Choi ),( Byoung Hyun Min ) 한국조직공학·재생의학회 2010 조직공학과 재생의학 Vol.7 No.1

This study investigated the effect of low intensity ultrasound (LIUS) stimulation of mesenchymal stem cells (MSCs) in vitro on the repair of cartilage defect after implantation of the construct in vivo. Rabbit MSCs were cultured in the polyglycolic acid (PGA) scaffold and preconditioned with (MSCs/US+) or without (MSCs/US-) LIUS stimulation during the chondrogenic differentiation for 1 week in vitro. The LIUS stimulation was carried out at the intensity of 200 mW/Cm2 every day for 20 min over a week. The constructs were implanted into the cartilage defects created in the rabbit femoral trochlea. The defect only was used as a negative control and rabbit chondrocytes seeded in PGA was used as a positive control, respectively. The repair of cartilage defect was examined at 2, 4, 8 and 12 weeks after implantation, respectively. The gross observation showed that the articular cartilage defects were filled with the repaired tissue in all groups. Histological and immunohistochemical analyses revealed, however, more intensive and widespread expression of proteoglycans and type II collagen in the MSCs/US+ group than in the MSCs/US- group. Fibrous tissues were observed mainly in the defect only group. The chondrocytes groups showed efficient repair of the defect by hyaline cartilage. In conclusion, this study suggested that LIUS preconditioning of MSCs in vitro could be an effective method to promote chondrogenesis of MSCs and repair of cartilage defect in vivo.

Effects of Low-Intensity Ultrasound on Chondrogenic Differentiation of Mesenchymal Stem Cells Embedded in Polyglycolic Acid: An <i>in Vivo</i> Study

Cui, Ji Hao,Park, Kwideok,Park, So Ra,Min, Byoung-Hyun Mary Ann Liebert 2006 Tissue engineering Vol.12 No.1

<P>In this study we investigated the effects of LIUS on chondrogenic differentiation of bone marrow-derived mesenchymal stem cells (BM-MSC). Our hypothesis is that LIUS may be a noninvasively effective stimulant to a biological system in vivo by turning on differentiation of MSCs and promotion of chondrogenesis. MSCs were isolated from the bone marrow of New Zealand white rabbits and cultured in monolayer for 2 weeks. They were then harvested and seeded into polyglycolic acid (PGA) non-woven mesh at a number of 5 x 10(6) cells. Cultured with a chondrogenic-defined media for 1 week, the PGA/MSCs constructs (n = 4) were implanted subcutaneously in the back of nude mice (n = 9, each group). The ultrasound (US) group received US stimulation at a frequency of 0.8 MHz and intensity of 200 mW/cm(2) for 10 min every day up to 4 weeks, while the control group had no US stimulation. Analyses of histological, immunohistochemical, biochemical, and mechanical characteristics were made at 1, 2, and 4 weeks post-stimulation, respectively. Total DNA contents showed no significant difference between the two groups. Total collagen and glycosaminoglycan (GAG) increased more significantly in the US-stimulated group than in the control. Histology of Safranin O/Fast green confirmed more intense and spreading extracellular matrix (ECM) at 2 and 4 weeks in the US-stimulated specimens. Mechanical tests exhibited that compressive strengths were also significantly higher in the US-stimulated cells at later times. This study strongly suggests that it may be possible for ultrasound to have some stimulatory effects in vivo on the chondrogenesis of MSCs.</P>

The Effects of Monolayer Subculture on the Phenotype and Growth of Degenerated Human Intervertebral Disc Cells

( Ji Hao Cui ),( Ho Geun Chang ),( Yong Chan Kim ) 한국조직공학·재생의학회 2012 조직공학과 재생의학 Vol.9 No.2

This study investigated the changes in the growth and phenotype of human degenerated intervertebral disc cells depending on the frequency of subculture in vitro monolayer culture system. The annulus fibrosus (AF) and nucleus pulposus (NP) cells were cultured in a monolayer manner. At each subculture , we performed an analysis of the morphological changes, the adhesion rate, the proliferation rate and the viability. The expressions of type I, II collagen and proteoglycan were analyzed at the mRNA gene level. All three times of subculture were performed. Both the AF and NP cells gradually showed a fibroblast-like spindle shape from their irregular, polygonal shape while undergoing the third time of subculture. The cell proliferation was the highest at the second subculture time. The viability was markedly lower before the first subculture, but there was no significant difference of viability between the three times of subculture. On RT-PCR, the type II collagen expression was gradually decreased and almost unexpressed at the third subculture, but the type I collagen expression was gradually increased in the NP cells. In the AF cells, the type I collagen expression did not show a significant difference depending on the frequency of subculture. Type II collagen was not expressed from the second time of subculture. The expression of proteoglycan was gradually decreased in both the AF and NP cells and it was almost unexpressed at the third time of subculture. In conclusions, after three times of subculture, the disc cells had completely changed their original growth and phenotypic characteristics.

The Combined Effects of Transforming Growth Factor-β and basic Fibroblast Growth Factor on the Human Degenerated Nucleus Pulposus Cells in Monolayer Culture

Erxing He,Ji Hao Cui,Chuang Li,Cheng Tang,김인성,김영우,장호근,차재룡,병오종,김용찬 한국조직공학과 재생의학회 2013 조직공학과 재생의학 Vol.10 No.3

This study investigated the changes in the growth and phenotype of human degenerated intervertebral disc cells depending on the frequency of subculture and the growth factors effects in vitro monolayer culture system. The twice passaged nucleus pulposus (NP) cells were replated six well plates at a density 1×104 cells/cm2. The replated NP cells were divided into four subgroups: no treatment (control group), the treatment of 10 ng/mL transforming growth factor (TGF)-β1 (TGF group), treatment of 5 ng/mL basic fibroblast growth factor (bFGF, FGF group), and treatment of TGF-β plus bFGF (TGF/FGF group). Analyses of proliferation, morphology, histological and reverse transcriptase - polymerase chain reaction (RT-PCR) were made at different time point. The NP cells gradually showed a fibroblast-like spindle shape from their irregular, polygonal shape while undergoing the third time of subculture. The cell proliferation was the highest at the second subculture time. On RT-PCR, the type II collagen expression was gradually decreased and almost unexpressed at the third subculture, but the type I collagen expression was gradually increased in the NP cells. The expression of proteoglycan was gradually decreased in NP cells and it was almost unexpressed at the third time of subculture. FGF can stimulate NP cell proliferation and TGF-β showed increase collagen type II and aggrecan synthesis, and the combination of TGF and FGF appeared both effects of proliferation and extracellular matrix (ECM) synthesis on the NP metabolism. To obtain a great number of cells using a small frequency of subculture and to increase the ECM synthesis, new culture methods based on growth factors or mechanical stimulation must be considered.

요추부 퇴행성질환 환자들의 시상면 정렬에 대한 골반의 역할

김용찬,유기한,장철영,Ji Hao Cui,Sirichai Wilartratsami,김태환,오재근,박문수,김석우 대한척추외과학회 2016 대한척추외과학회지 Vol.23 No.2

연구 계획: 종설. 목적: 골반과 요추부 퇴행성질환 환자들의 상호 관계에 대한 최신 지견을 제시하고, 요추부 퇴행성질환 환자들의 시상면 정렬에 대한 골반의 역할을 강조하고자 한다. 선행 문헌의 요약: 골반과 요추부 퇴행성질환 환자들의 시상면 정렬간의 관계에 대해서는 논쟁의 여지가 있지만, 많은 저자들은 골반이 요추부 퇴행성 질환 환자들의 시상면 정렬에 대해서 의미 있는 영향을 미친다고 보고해 왔다. 대상 및 방법: 저자들은 골반과 요추부 퇴행성질환 환자들에 대한 참고 문헌들을 검색하였고, 지난 30년간의 문헌에 대해 꾸준히 추적 관찰하였다. 결과: 골반과 요추간의 관계는 전 인체 시상면 정렬에 영향을 미친다. 또한 골반과 요추부 퇴행성질환 환자들의 시상면 정렬간에는 매우 밀접한 상호관계가 있다. 따라서 골반과 요추는 요추골반복합체(Lumbopelvic complex)라는 하나의 개념으로 간주되어야 한다. 결론: 우리는 요천추관절(Lumbosacral joint)이 아닌 요추골반관절(Lumbopelvic joint)의 개념을, 그리고 요추전만(Lumbar lordosis)이 아닌 요추골반전만(Lumbopelvic lordosis)의 개념을 가질 필요가 있다. 또한 전 인체 시상면 정렬과 동적 균형을 평가하기 위하여 요추골반복합체를 고려해야만 한다. Study Design: Literature review. Objectives: To present updated information on the relationship of the pelvis and lumbar degenerative disease (LDD) patients and to emphasize the importance of the pelvis in sagittal alignment of LDD patients. Summary of Literature Review: Although the relationship of the pelvis and sagittal alignment of LDD patients is controversial, many authors have reported a significant impact of the pelvis on LDD sagittal alignment. Materials and Methods: The authors identified references through a literature search on the pelvis and LDD and continuous monitoring of the literature during the past 30 years. Results: The pelvis and lumbar levels were related to whole-body sagittal alignment. The pelvis is also closely related to sagittal alignment of LDD patients. Therefore, the entire area should be regarded as a lumbopelvic complex. Conclusions: We need to consider the concept of a lumbopelvic joint and lumbopelvic lordosis, not a lumbosacral joint and lumbar lordosis. We must also evaluate the lumbopelvic complex to assess whole-body sagittal alignment and dynamic balance.

Potential of Fortified Fibrin/Hyaluronic Acid Composite Gel as a Cell Delivery Vehicle for Chondrocytes

Park, Sang-Hyug,Cui, Ji Hao,Park, So Ra,Min, Byoung-Hyun Blackwell Publishing Inc 2009 Artificial Organs Vol.33 No.6

<P>Abstract</P><P>Numerous treatment methods have been applied for use in cartilage repair, including abrasion, drilling, and microfracture. Although chondrocyte transplantation is the preferred treatment, it has some shortcomings, such as difficulty of application (large and posterior condylar regions), poor chondrocyte distribution, and potential cell leakage from the defect region. The cell delivery system of the tissue engineering technique can be used to overcome these shortcomings. We chose fibrin/hyaluronan (HA) composite gel as an effective cell delivery system to resolve these issues. Both components are derived from natural extracellular matrix. In the first trial, fortified fibrin/HA composite gels with rabbit chondrocytes were tested by implantation in nude mice. At 4 weeks, glycosaminoglycan contents in the fibrin/HA composite (0.186 ± 0.006 mg/mg) were significantly higher than those in the presence of fibrin alone (0.153 ± 0.017 mg/mg). As a next step, we applied the fibrin/HA composite gel to animal cartilage defects using full thickness cartilage defect rabbit models. The fibrin/HA composite gel with rabbit chondrocytes (allogenic) was implanted into the experimental group, and the control group was implanted with the fibrin/HA composite gel alone. Implanted chondrocytes with the fibrin/HA composite showed improved cartilage formation. In conclusion, the key to successful regeneration of cartilage is to provide the repair site with a sufficient supply of chondrogenic cells with a suitable delivery vehicle to ensure maximal differentiation and deposition of the proper extracellular matrix. This study suggests the feasibility of tissue-engineered cartilage formation using fibrin/HA composite gel.</P>

Ethanol Extract of <i>Lepidium apetalum</i> Seed Elicits Contractile Response and Attenuates Atrial Natriuretic Peptide Secretion in Beating Rabbit Atria

Kim, Seung Ju,Kim, Hye Yoom,Lee, Yun Jung,Cui, Hao Zhen,Jang, Ji Yeon,Kang, Dae Gill,Lee, Ho Sub Hindawi Publishing Corporation 2013 Evidence-based Complementary and Alternative Medic Vol.2013 No.-

<P>The seeds of <I>Lepidium apetalum</I> Willdenow (called “Tinglizi” in China and “Jungryukza” in Korea) have been used to discharge phlegm and improve dropsy in Oriental medicine. The present study investigated the effects of ethanol extract of the seeds of <I>Lepidium apetalum</I> (ELA) on atrial dynamics and atrial natriuretic peptide (ANP) secretion in beating rabbit atria. ELA increased atrial stroke volume, pulse pressure, and cAMP efflux, concomitantly attenuating ANP secretion in a dose-dependent manner. ELA-induced increases in atrial stroke volume, pulse pressure, and cAMP levels and decrease in ANP secretion were not inhibited by pretreatment with staurosporine, a nonspecific protein kinase inhibitor, or diltiazem and verapamil, the L-type Ca<SUP>2+</SUP> channel blockers, respectively. Helveticoside, a well-known digitalis-like cardiac glycosidic constituent of ELA, also increased atrial dynamics, including stroke volume and pulse pressure, without changing cAMP efflux and ANP secretion, and the effects of helveticoside were not inhibited by pretreatment with staurosporine, diltiazem, and verapamil. These results suggest that the ELA-induced positive inotropic activity in beating rabbit atria might, at least partly, be due to the digitalis-like activity of helveticoside rather than an increase in cAMP efflux.</P>