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Matrix replenish is the initial requirement of the osteoarthritic cartilage
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Abstracts / Osteoarthritis and Cartilage 25 (2017) S76eS444
242 CARTILAGE REPAIR BY VARIOUS CONCENTRATIONS OF PLACENTADERIVED MESENCHYMAL STEM CELLS AND HYALURONIC ACID HYDROGELS IN A RABBIT MODEL J. Rhim y, C.-W. Ha y, Y.-B. Park z, J.-A. Kim y, W.-J. Han y, S. Choi y, K. Lee y, H. Park y, H.-J. Park y. y Samsung Med. Ctr., Stem Cell & Regenerative Med. Res. Ctr., Sungkyunkwan Univ. Sch. of Med., Seoul, Republic of Korea; z Chung-Ang Univ. Hosp., Chung-Ang Univ. Coll. of Med., Seoul, Republic of Korea Purpose: The placenta-derived mesenchymal stem cells have great interest as a promising cell source for regenerative medicine due to non-invasive collection, readily availability, high expansion capacity, and low immunogenicity. We investigated the feasibility of transplanting placenta-derived MSCs and a hyaluronic acid (HA) hydrogel composite to repair full-thickness articular cartilage defects and determine whether the transplantation of various concentrations of placenta-derived MSCs with 4% hyaluronic acid (HA) hydrogel promoted articular cartilage repair in a rabbit model. Methods: Critical-sized osteochondral defects (5 mm wide 5 mm deep) were created in the center of the trochlear groove of femur. The Four experimental groups were as follows: group A, defect only (left knee)/ 4% HA hydrogel without placenta-MSCs (right knee), n ¼ 6; group B, 0.01 x 107 cells/ml with 4% HA hydrogel (left knee)/ 0.1 x 107 cells/ml with 4% HA hydrogel (right knee), n ¼ 8; group C, 0.1 x 107 cells/ ml with 4% HA hydrogel (left knee)/ 0.5 x 107 cells/ml with 4% HA hydrogel (right knee), n ¼ 8; group D, 0.5 x 107 cells/ml with 4% HA hydrogel (left knee)/ 1 x 107 cells/ml with 4% HA hydrogel (right knee) were transplantation in a rabbit model. At 8 weeks after transplantation, the degree of cartilage repair was evaluated grossly and histologically using the International Cartilage Repair Society macroscopic score, the modified O’Driscoll score for histologic grading. Results: Transplanting placenta-derived MSCs and a HA hydrogel composite resulted in overall superior cartilage repair tissue with better quality than no treatment and hyaluronic acid (HA) only treatment group. Interestingly, The 0.5 x 106 cells/ml group showed the highest cartilage repair at 8 weeks post transplantation. Conclusions: The results of this study suggest that transplantation of the composite of placenta-derived MSCs and HA is promoting for cartilage repair. In addition, this study shows that optimal MSC concentration needs to be determined for better cartilage repair. 243 MATRIX REPLENISH IS THE INITIAL REQUIREMENT OF THE OSTEOARTHRITIC CARTILAGE M. Zhang y, X. Wan y, L. Lu y, H. Yang y, J. Zhang y, H. Zhang y, X. Liu y, X. Huang z, G. Xiao x, M. Wang k. y Coll. of Somatology, Fourth MIlitary Med. Univ., Xi'an, China; z Dept. Biology, Fourth MIlitary Med. Univ., Xi'an, China; x Dept. of Biology and Shenzhen Key Lab. of Cell Microenvironment, South Univ. of Sci. and Technology of China, Shenzhen, China; k Coll. of Somatology, Fourth MIlitary Med. Univ., Xian, China Purpose: The initial requirement of the osteoarthritic (OA) cartilage is vitally important. Understanding of it will bring the insight of disease process and thus will be helpful for an effective therapy of OA. Temporomandibular joint (TMJ) is one of the sites that are often insulted by OA due to the aberrant biomechanical stress. Our previous reports indicated that an experimental dental stimulation termed unilateral anterior crossbite (UAC) elicited OA lesions in rodent TMJs, including not only the loss of subchondral bone and enhancement of the osteochondral interference calcification, but also apoptosis and necrosis of the chondrocytes and losing of the cartilage matrix. Up-regulated phagocytosis of the chondrocytes has been identified in OA cartilage. Local injected green fluorescent protein labeled exogenous bone marrow stromal cells (GFP-BMSCs), which have a high capability of replication, were attracted to the insulted sites of UAC induced TMJ OA cartilage via SDF-1/CXCR4 and RANTES/CCR1 signalings. The GFPBMSCs were then implanted in the degraded cartilage and differentiated to chondrocytes due to which the matrix was supplemented
and the degraded cartilage was restored. However, what is the primary process of the lesion and what is initially required for restoration of the OA cartilage is still obscure. The present aim was to explore whether the matrix replenish or the cellular replica is initially required by OA cartilage. Methods: UAC was applied to C57BL/6J female mice (17-19 g), 6 weeks of age. GFP-BMSCs were weekly injected into TMJs, stating from 3 weeks of UAC stimulation, for 4, 8 and 12 weeks. Another GFP-BMSCs injection UAC group was stop to be injected at 8 weeks and were continued to be raised for another 4 weeks. Alterations in TMJ cartilage were determined at the molecular, cellular, and tissue levels through histochemistrical, immunohistochemistrical or double fluorescent staining, transmission electron microscope (TEM) observation, western blot or realtime PCR detection. UAC effects on TMJ chondrocyte anoikis, scavenging activity and proliferation were assessed by measuring the expression levels of DAP3, an anoikis marker, CD163, a scavenger receptor family member, and ki67, a proliferation indicator, respectively. Difference of the cellular responses from superficial and deep zone cartilage to the shearing stress was also detected by isolating chondrocytes from superficial and deep zone of pig TMJ cartilage and exposing them to the fluid flow shear stress (FFSS) flow for 1 h at intensities of 4, 12 or 24 dyn/cm2. Results: Matrix losing and thickness reduction appeared significant in deep zones of the TMJ cartilage obtained from UAC mice (Figure 1a-f). For superficial zone, however, fibrillation was obvious at late stage (Fig.1g, h). DAP3 expression was upregulated which indicated a cellular death of anoikiss. Meanwhile, there was an increasing of CD163 expression in chondrocytes implying an enhanced scavenging activity. However, cell proliferation was not observable. The expression level of ki67 was not changed, though cellular aggregates were formed from early stage. Weekly injection of GFP-BMSCs largely restored the degradation (Figure 1i-k). DAP3 expression level was reduced and CD163 protein was expressed by the implanted GFP-BMSCs. However, there were no signs of increasing of cell proliferation. The ki67 expression level was unchanged throughout the observing period. Impressively, stop supply of the GFP-BMSCs ruined the restoring effects, leading to cartilage decay relapse (Figure 1l). In vitro data indicated that FFSS upregulated fibrous matrix, type I collagen, but down-regulated cartilage matrix, aggrecan, in cells isolated from both superficial and deep zones. Conclusions: Deep zone decay of the TMJ cartilage was induced by UAC but in the superficial zone fibrillation was the predominance. Scavenging the degraded cartilage is one of the principle attempts of chondrocytes in progressive TMJ OA cartilage though usually unsuccessful. Continuing supply of the exogenous GFP-BMSCs satisfied the requirements of matrix replenish via scavenging and matrix producing rather than cell replica. Considerring that BMSCs could find the way to behavior as the target tissue required, we come to the conclusion that matrix replenish is the initial requirement of the biomechanically induced TMJ osteoarthritis.
Abstracts / Osteoarthritis and Cartilage 25 (2017) S76eS444
242 CARTILAGE REPAIR BY VARIOUS CONCENTRATIONS OF PLACENTADERIVED MESENCHYMAL STEM CELLS AND HYALURONIC ACID HYDROGELS IN A RABBIT MODEL J. Rhim y, C.-W. Ha y, Y.-B. Park z, J.-A. Kim y, W.-J. Han y, S. Choi y, K. Lee y, H. Park y, H.-J. Park y. y Samsung Med. Ctr., Stem Cell & Regenerative Med. Res. Ctr., Sungkyunkwan Univ. Sch. of Med., Seoul, Republic of Korea; z Chung-Ang Univ. Hosp., Chung-Ang Univ. Coll. of Med., Seoul, Republic of Korea Purpose: The placenta-derived mesenchymal stem cells have great interest as a promising cell source for regenerative medicine due to non-invasive collection, readily availability, high expansion capacity, and low immunogenicity. We investigated the feasibility of transplanting placenta-derived MSCs and a hyaluronic acid (HA) hydrogel composite to repair full-thickness articular cartilage defects and determine whether the transplantation of various concentrations of placenta-derived MSCs with 4% hyaluronic acid (HA) hydrogel promoted articular cartilage repair in a rabbit model. Methods: Critical-sized osteochondral defects (5 mm wide 5 mm deep) were created in the center of the trochlear groove of femur. The Four experimental groups were as follows: group A, defect only (left knee)/ 4% HA hydrogel without placenta-MSCs (right knee), n ¼ 6; group B, 0.01 x 107 cells/ml with 4% HA hydrogel (left knee)/ 0.1 x 107 cells/ml with 4% HA hydrogel (right knee), n ¼ 8; group C, 0.1 x 107 cells/ ml with 4% HA hydrogel (left knee)/ 0.5 x 107 cells/ml with 4% HA hydrogel (right knee), n ¼ 8; group D, 0.5 x 107 cells/ml with 4% HA hydrogel (left knee)/ 1 x 107 cells/ml with 4% HA hydrogel (right knee) were transplantation in a rabbit model. At 8 weeks after transplantation, the degree of cartilage repair was evaluated grossly and histologically using the International Cartilage Repair Society macroscopic score, the modified O’Driscoll score for histologic grading. Results: Transplanting placenta-derived MSCs and a HA hydrogel composite resulted in overall superior cartilage repair tissue with better quality than no treatment and hyaluronic acid (HA) only treatment group. Interestingly, The 0.5 x 106 cells/ml group showed the highest cartilage repair at 8 weeks post transplantation. Conclusions: The results of this study suggest that transplantation of the composite of placenta-derived MSCs and HA is promoting for cartilage repair. In addition, this study shows that optimal MSC concentration needs to be determined for better cartilage repair. 243 MATRIX REPLENISH IS THE INITIAL REQUIREMENT OF THE OSTEOARTHRITIC CARTILAGE M. Zhang y, X. Wan y, L. Lu y, H. Yang y, J. Zhang y, H. Zhang y, X. Liu y, X. Huang z, G. Xiao x, M. Wang k. y Coll. of Somatology, Fourth MIlitary Med. Univ., Xi'an, China; z Dept. Biology, Fourth MIlitary Med. Univ., Xi'an, China; x Dept. of Biology and Shenzhen Key Lab. of Cell Microenvironment, South Univ. of Sci. and Technology of China, Shenzhen, China; k Coll. of Somatology, Fourth MIlitary Med. Univ., Xian, China Purpose: The initial requirement of the osteoarthritic (OA) cartilage is vitally important. Understanding of it will bring the insight of disease process and thus will be helpful for an effective therapy of OA. Temporomandibular joint (TMJ) is one of the sites that are often insulted by OA due to the aberrant biomechanical stress. Our previous reports indicated that an experimental dental stimulation termed unilateral anterior crossbite (UAC) elicited OA lesions in rodent TMJs, including not only the loss of subchondral bone and enhancement of the osteochondral interference calcification, but also apoptosis and necrosis of the chondrocytes and losing of the cartilage matrix. Up-regulated phagocytosis of the chondrocytes has been identified in OA cartilage. Local injected green fluorescent protein labeled exogenous bone marrow stromal cells (GFP-BMSCs), which have a high capability of replication, were attracted to the insulted sites of UAC induced TMJ OA cartilage via SDF-1/CXCR4 and RANTES/CCR1 signalings. The GFPBMSCs were then implanted in the degraded cartilage and differentiated to chondrocytes due to which the matrix was supplemented
and the degraded cartilage was restored. However, what is the primary process of the lesion and what is initially required for restoration of the OA cartilage is still obscure. The present aim was to explore whether the matrix replenish or the cellular replica is initially required by OA cartilage. Methods: UAC was applied to C57BL/6J female mice (17-19 g), 6 weeks of age. GFP-BMSCs were weekly injected into TMJs, stating from 3 weeks of UAC stimulation, for 4, 8 and 12 weeks. Another GFP-BMSCs injection UAC group was stop to be injected at 8 weeks and were continued to be raised for another 4 weeks. Alterations in TMJ cartilage were determined at the molecular, cellular, and tissue levels through histochemistrical, immunohistochemistrical or double fluorescent staining, transmission electron microscope (TEM) observation, western blot or realtime PCR detection. UAC effects on TMJ chondrocyte anoikis, scavenging activity and proliferation were assessed by measuring the expression levels of DAP3, an anoikis marker, CD163, a scavenger receptor family member, and ki67, a proliferation indicator, respectively. Difference of the cellular responses from superficial and deep zone cartilage to the shearing stress was also detected by isolating chondrocytes from superficial and deep zone of pig TMJ cartilage and exposing them to the fluid flow shear stress (FFSS) flow for 1 h at intensities of 4, 12 or 24 dyn/cm2. Results: Matrix losing and thickness reduction appeared significant in deep zones of the TMJ cartilage obtained from UAC mice (Figure 1a-f). For superficial zone, however, fibrillation was obvious at late stage (Fig.1g, h). DAP3 expression was upregulated which indicated a cellular death of anoikiss. Meanwhile, there was an increasing of CD163 expression in chondrocytes implying an enhanced scavenging activity. However, cell proliferation was not observable. The expression level of ki67 was not changed, though cellular aggregates were formed from early stage. Weekly injection of GFP-BMSCs largely restored the degradation (Figure 1i-k). DAP3 expression level was reduced and CD163 protein was expressed by the implanted GFP-BMSCs. However, there were no signs of increasing of cell proliferation. The ki67 expression level was unchanged throughout the observing period. Impressively, stop supply of the GFP-BMSCs ruined the restoring effects, leading to cartilage decay relapse (Figure 1l). In vitro data indicated that FFSS upregulated fibrous matrix, type I collagen, but down-regulated cartilage matrix, aggrecan, in cells isolated from both superficial and deep zones. Conclusions: Deep zone decay of the TMJ cartilage was induced by UAC but in the superficial zone fibrillation was the predominance. Scavenging the degraded cartilage is one of the principle attempts of chondrocytes in progressive TMJ OA cartilage though usually unsuccessful. Continuing supply of the exogenous GFP-BMSCs satisfied the requirements of matrix replenish via scavenging and matrix producing rather than cell replica. Considerring that BMSCs could find the way to behavior as the target tissue required, we come to the conclusion that matrix replenish is the initial requirement of the biomechanically induced TMJ osteoarthritis.