脂肪間葉幹細胞(adipose-derived mesenchymal stem cells, ASCs)為多潛能型幹細胞，具有分化成軟骨細胞之潛力且容易自脂肪組織分離，現已廣泛應用於組織工程。然而，脂肪間葉幹細胞的軟骨分化效率與骨髓間葉幹細胞相比卻明顯不足。我們假設若能將刺激ASCs軟骨化的生長因子濃度增加，並培養於低氧氣濃度以及動態培養環境下，將有利於ASCs分化為軟骨細胞(chondrocytes)，並且在體外培養時形成工程軟骨(engineered cartilage)。 因此，本研究之初期目的便是利用重組桿狀病毒(Bac-CT3W)表現轉化生長因子(transforming growth factor-β3, TGF-β3)來改質ASCs，並培養於兩相旋轉軸生物反應器(rotating shaft bioreactor, RSB)中，期望結合生物、環境與物理刺激能夠促進ASCs分化成軟骨細胞，並形成工程軟骨。我們證實經由Bac-CT3W轉導過的ASCs有劑量依賴性且能夠表現高濃度的TGF-β3生長因子。基因改質後的ASCs接種於聚乳酸-甘醇酸(Poly(Lactide-co-Glycolide), PLGA)細胞支架並在RSB反應器中培養兩週後，細胞形態類似軟骨細胞，並可以在細胞/支架結構物的表面及周圍產生大量的軟骨特異性細胞外間質，如葡萄糖胺聚醣(glycosaminoglycans, GAGs)與第二型膠原蛋白(collagen type II, COL II)。當培養週數超過兩週，結構物的細胞外基質會減少，可能是因為細胞外基質的分子分解所導致。總結來說，結合重組桿狀病毒基因改質的ASCs與RSB的動態培養能夠提高ASCs的軟骨分化效果並形成類軟骨組織，但培養時間卻是形成優化的工程軟骨之重要關鍵因素。
由於ASCs於體外分化緩慢，因此我們假設若能對ASCs長期提供生長因子刺激以利於體外培養形成軟骨組織，將有利於培養出更接近天然軟骨組織之工程軟骨並加速其體外培養的時間和加強工程軟骨的體外培養效果。因此，我們利用能夠長效表現TGF-β3和第六骨形態發生蛋白質(bone morphological protein-6, BMP-6)的混成桿狀病毒來改質ASCs，希望利用具有抑制軟骨細胞肥大化(hypertrophy)能力的TGF-β3和能增強ASCs細胞表面之轉化生長因子受體-I (TGF-β receptor I)表現的BMP-6之協同作用，增強刺激ASCs往軟骨化路徑分化的效果。我們將基因改質後的ASCs植入已由明膠(gelatin)、透明質酸(hyaluronic acid, HA)和硫酸軟骨素(chondroitin sulfate, CS)共聚合接枝在PLGA支架上而成的PLGA-GCH細胞支架中，並培養於接近天然軟骨環境之低氧氣濃度條件。實驗結果證實，經由混成長效型重組桿狀病毒表現系統改質之ASCs在3D立體培養環境下能維持混成生長因子群之高濃度，刺激ASCs往軟骨細胞路徑分化並表現軟骨標誌基因(SOX 9、Aggrecan和COL2A1)，抑制纖維軟骨標誌基因(COL1A2)與細胞肥大化標誌基因(COL10A1)之表現，且工程軟骨產生大量的軟骨特異性細胞外間質如GAGs與COL II。體外培養兩週之工程軟骨在植入紐西蘭白兔的膝關節負重缺陷處後，以磁振造影(magnetic resonance imaging, MRI)能清楚的觀察到以混成長效型表現系統培養之工程軟骨能在術後12週即達到良好的軟骨組織修復效果。本研究是首次結合新穎的長效型表現生長因子之重組桿狀病毒改質的ASCs與PLGA-GCH細胞支架，並於體外環境下培養出工程軟骨，應用於兔膝關節負重區之軟骨組織修復，本實驗結果證實混成長效型表現系統於軟骨組織工程領域具有高度的研究潛力且具有未來臨床應用價值。

Adipose-derived mesenchymal stem cells (ASCs) are easy to isolate from adipose tissues in abundance and have been widely employed in tissue engineering. However, ASCs appear to be inferior to bone marrow mesenchymal stem cells in terms of chondrogenic potential. We hypothesize that proper supply of transforming growth factor, low oxygen concentration and a dynamic environment could facilitate ASCs to differentiate into chondrocytes and to promote the engineered cartilage formation. Therefore, in this study we first developed a baculoviral vector that expressed TGF-β3 (Bac-CT3W) for the genetic modification of ASCs, which were then cultured in the two-phase rotating shaft bioreactor (RSB), hoping that the biological, environmental and physical stimuli altogether can facilitate the ASCs differentiation into chondrocytes and growth into engineered cartilages in vitro. We demonstrated that ASCs transduced with Bac-CT3W expressed high levels of TGF-β3 in a dose-dependent manner. After seeding the cells into 3D, porous scaffolds and culture in the RSB for 2 weeks, the cell/scaffold constructs grew into cartilage-like tissues with hyaline appearance and the cells resembled the authentic chondrocytes. The constructs were abundant in cartilage-specific extracellular matrix (ECM) including collagen II and glycosaminoglycan as demonstrated by histological staining and biochemical assays. Intriguingly, when the construct culture time exceeded 2 weeks, the abundance of ECM decreased, probably due to the degradation of the ECM molecules. These data demonstrated that combining the TGF-β3-expressing baculovirus and RSB for the genetic modification and dynamic culture of ASC enhances the chondrogenic differentiation of ASCs and cartilage formation, but the culture time appears to be critical to the cartilage quality.
The chondrogenic differentiation of ASCs is time-consuming and requires sustained growth factor expression. We further developed hybrid baculoviruses that can persistently express TGF-β3 and bone morphological protein-6 (BMP-6) to genetically modify ASCs, hoping that TGF-β3 can suppress the hypertrophy of chondrocytes while BMP-6 can further enhance the chondrogenesis of ASCs. Furthermore, we developed new scaffolds that comprised PLGA, gelatin, chondroitin sulfate and hyaluronic acid (PLGA-GCH), hoping that these components futher synergistically promote ASCs chondrogenesis. The genetically engineered ASCs were seeded to the porous PLGA-GCH scaffolds and cultivated under low oxygen concentration condition that mimic the native cartilage environment. We demonstrated that transduction of ASCs with the hybrid baculovirus enabled sustained expression of TGF-β3 and BMP-6 at high levels in the 3D environment, stimulated the chondrogenesis of ASCs and formation of cartilage-like tissues. Transplantation of the engineered constructs into the osteochondral defects at the weight-bearing area of medial femoral condyle of rabbit knees accelerated and ameliorated the cartilage repair in 12 weeks, as evidenced by MRI. This study, for the first time, combines PLGA-GCH scaffolds and engineered ASCs persistently expressing grwoth factors for the formation of engineered cartilages and in vivo repair of osteochondral defects.

Abe T, Hemmi H, Miyamoto H, Moriishi K, Tamura S, Takaku H, Akira S, Matsuura Y. 2005. Involvement of the Toll-like receptor 9 signaling pathway in the induction of innate immunity by baculovirus. J Virol 79(5):2847-58.
Abramson SB. 2008. Nitric oxide in inflammation and pain associated with osteoarthritis. Arthritis Res Ther 10 Suppl 2:S2.
Afizah H, Yang Z, Hui JH, Ouyang HW, Lee EH. 2007. A comparison between the chondrogenic potential of human bone marrow stem cells (BMSCs) and adipose-derived stem cells (ADSCs) taken from the same donors. Tissue Eng 13(4):659-66.
Alford JW, Cole BJ. 2005. Cartilage restoration, part 1: basic science, historical perspective, patient evaluation, and treatment options. Am J Sports Med 33(2):295-306.
Arikawa T, Matsukawa A, Watanabe K, Sakata K-m, Seki M, Nagayama M, Takeshita K, Ito K, Niki T, Oomizu S. 2009. Galectin-9 accelerates transforming growth factor β3-induced differentiation of human mesenchymal stem cells to chondrocytes. Bone 44(5):849-857.
Barry F, Boynton RE, Liu B, Murphy JM. 2001. Chondrogenic differentiation of mesenchymal stem cells from bone marrow: differentiation-dependent gene expression of matrix components. Exp Cell Res 268(2):189-200.
Baum C. 2007. What are the consequences of the fourth case? Mol. Ther. 15:1401-1402.
Bilello JP, Cable EE, Myers RL, Isom HC. 2003. Role of paracellular junction complexes in baculovirus-mediated gene transfer to nondividing rat hepatocytes. Gene Ther 10(9):733-49.
Bouffi C, Thomas O, Bony C, Giteau A, Venier-Julienne M-C, Jorgensen C, Montero-Menei C, Noël D. 2010. The role of pharmacologically active microcarriers releasing TGF-β3 in cartilage formation in vivo by mesenchymal stem cells. Biomaterials 31(25):6485-6493.
Broll S, Oumard A, Hahn K, Schambach A, Bode J. 2010. Minicircle performance depending on S/MAR-nuclear matrix interactions. J Mol Biol 395(5):950-65.
Burkhardt H, Schwingel M, Menninger H, Macartney HW, Tschesche H. 1986. Oxygen radicals as effectors of cartilage destruction. Direct degradative effect on matrix components and indirect action via activation of latent collagenase from polymorphonuclear leukocytes. Arthritis Rheum 29(3):379-87.
Candrian C, Miot S, Wolf F, Bonacina E, Dickinson S, Wirz D, Jakob M, Valderrabano V, Barbero A, Martin I. 2010. Are ankle chondrocytes from damaged fragments a suitable cell source for cartilage repair? Osteoarthritis and Cartilage 18(8):1067-1076.
Cao M, Westerhausen-Larson A, Niyibizi C, Kavalkovich K, Georgescu HI, Rizzo CF, Hebda PA, Stefanovic-Racic M, Evans CH. 1997. Nitric oxide inhibits the synthesis of type-II collagen without altering Col2A1 mRNA abundance: prolyl hydroxylase as a possible target. Biochem J 324 ( Pt 1):305-10.
Chang J-C, Hsu S-h, Chen DC. 2009. The promotion of chondrogenesis in adipose-derived adult stem cells by an RGD-chimeric protein in 3D alginate culture. Biomaterials 30(31):6265-6275.
Chang JK, Ho ML, Lin SY. 1996. Effects of compressive loading on articular cartilage repair of knee joint in rats. Kaohsiung J Med Sci 12(8):453-60.
Chen H-C, Chang Y-H, Chuang C-K, Lin C-Y, Sung L-Y, Wang Y-H, Hu Y-C. 2009a. The repair of osteochondral defects using baculovirus-mediated gene transfer with de-differentiated chondrocytes in bioreactor culture. Biomaterials 30:674-681.
Chen H-C, Lee H-P, Ho Y-C, Sung M-L, Hu Y-C. 2006a. Combination of baculovirus-mediated gene transfer and rotating-shaft bioreactor for cartilage tissue engineering. Biomaterials 27(16):3154-3162.
Chen H-C, Sung L-Y, Lo W-H, Chuang C-K, Wang Y-H, Lin J-L, Hu Y-C. 2008a. Combination of baculovirus-mediated BMP-2 expression and rotating-shaft bioreactor culture synergistically enhances cartilage formation. Gene Ther. 15:309-317.
Chen HC, Chang YH, Chuang CK, Lin CY, Sung LY, Wang YH, Hu YC. 2009b. The repair of osteochondral defects using baculovirus-mediated gene transfer with de-differentiated chondrocytes in bioreactor culture. Biomaterials 30(4):674-81.
Chen S-J, Lin C-C, Tuan W-C, Tseng C-S, Huang R-N. 2009. Effect of recombinant galectin-1 on the growth of immortal rat chondrocyte on chitosan-coated PLGA scaffold. Journal of Biomedical Materials Research Part A:n/a-n/a.
Choi S-J, Na K, Kim S, Woo DG, Sun BK, Chung H-M, Park K-H. 2007. Combination of ascorbate and growth factor (TGF β-3) in thermo-reversible hydrogel constructs embedded with rabbit chondrocytes for neocartilage formation. Journal of Biomedical Materials Research Part A 83A(4):897-905.
Clancy RM, Amin AR, Abramson SB. 1998. The role of nitric oxide in inflammation and immunity. Arthritis Rheum 41(7):1141-51.
Cucchiarini M, Madry H. 2005. Gene therapy for cartilage defects. J. Gene Med. 7(12):1495-1509.
Cui L, Wu Y, Cen L, Zhou H, Yin S, Liu G, Liu W, Cao Y. 2009. Repair of articular cartilage defect in non-weight bearing areas using adipose derived stem cells loaded polyglycolic acid mesh. Biomaterials 30(14):2683-2693.
Danisovic L, Varga I, Polak S. 2012. Growth factors and chondrogenic differentiation of mesenchymal stem cells. Tissue Cell 44(2):69-73.
Diekman BO, Estes BT, Guilak F. 2010. The effects of BMP6 overexpression on adipose stem cell chondrogenesis: Interactions with dexamethasone and exogenous growth factors. J Biomed Mater Res A 93(3):994-1003.
Donsante A, Miller DG, Li Y, Vogler C, Brunt EM, Russell DW, Sands MS. 2007. AAV vector integration sites in mouse hepatocellular carcinoma. Science 317(5837):477-477.
du Souich P, Garcia AG, Verges J, Montell E. 2009. Immunomodulatory and anti-inflammatory effects of chondroitin sulphate. J Cell Mol Med 13(8A):1451-63.
Dwarakanath RS, Clark CL, McElroy AK, Spector DH. 2001. The use of recombinant baculoviruses for sustained expression of human cytomegalovirus immediate early proteins in fibroblasts. Virology 284(2):297-307.
Elvenes J, Knutsen G, Johansen O, Moe BT, Martinez I. 2009. Development of a new method to harvest chondroprogenitor cells from underneath cartilage defects in the knees. Journal of Orthopaedic Science 14(4):410-417.
Enobakhare BO, Bader DL, Lee DA. 1996. Quantification of sulfated glycosaminoglycans in chondrocyte/alginate cultures, by use of 1,9-dimethylmethylene blue. Anal Biochem 243(1):189-91.
Erickson GR, Gimble JM, Franklin DM, Rice HE, Awad H, Guilak F. 2002. Chondrogenic potential of adipose tissue-derived stromal cells in vitro and in vivo. Biochem Biophys Res Commun 290(2):763-9.
Estes BT, Wu AW, Guilak F. 2006a. Potent induction of chondrocytic differentiation of human adipose-derived adult stem cells by bone morphogenetic protein 6. Arthritis Rheum 54(4):1222-32.
Estes BT, Wu AW, Storms RW, Guilak F. 2006b. Extended passaging, but not aldehyde dehydrogenase activity, increases the chondrogenic potential of human adipose-derived adult stem cells. Journal of Cellular Physiology 209(3):987-995.
Fan H, Tao H, Wu Y, Hu Y, Yan Y, Luo Z. 2010. TGF-β3 immobilized PLGA-gelatin/chondroitin sulfate/hyaluronic acid hybrid scaffold for cartilage regeneration. Journal of Biomedical Materials Research Part A 95A(4):982-992.
Feng G, Wan Y, Balian G, Laurencin CT, Li X. 2008. Adenovirus-mediated expression of growth and differentiation factor-5 promotes chondrogenesis of adipose stem cells. Growth Factors 26(3):132-42.
Furukawa T, Eyre DR, Koide S, Glimcher MJ. 1980. Biochemical studies on repair cartilage resurfacing experimental defects in the rabbit knee. J Bone Joint Surg Am 62(1):79-89.
Gelse K, Schneider H. 2006. Ex vivo gene therapy approaches to cartilage repair. Adv. Drug Deliv. Rev. 58(2):259-284.
Girotto D, Urbani S, Brun P, Renier D, Barbucci R, Abatangelo G. 2003. Tissue-specific gene expression in chondrocytes grown on three-dimensional hyaluronic acid scaffolds. Biomaterials 24(19):3265-75.
Gomes RR, Jr., Farach Carson MC, Carson DD. 2003. Perlecan-stimulated nodules undergo chondrogenic maturation in response to rhBMP-2 treatment in vitro. Connect Tissue Res 44 Suppl 1:196-201.
Han Y, Wei Y, Wang S, Song Y. 2010. Cartilage regeneration using adipose-derived stem cells and the controlled-released hybrid microspheres. Joint Bone Spine 77(1):27-31.
Helmers S, Sharkey PF, McGuigan FX. 1999. Efficacy of irrigation for removal of particulate debris after cemented total knee arthroplasty. J Arthroplasty 14(5):549-52.
Hennig T, Lorenz H, Thiel A, Goetzke K, Dickhut A, Geiger F, Richter W. 2007. Reduced chondrogenic potential of adipose tissue derived stromal cells correlates with an altered TGFbeta receptor and BMP profile and is overcome by BMP-6. J Cell Physiol 211(3):682-91.
Ho Y-C, Chen H-C, Wang K-C, Hu Y-C. 2004. Highly efficient baculovirus-mediated gene transfer into rat chondrocytes. Biotechnol. Bioeng. 88(5):643-51.
Ho Y-C, Chung Y-C, Hwang S-M, Wang K-C, Hu Y-C. 2005. Transgene expression and differentiation of baculovirus-transduced human mesenchymal stem cells. J. Gene. Med. 7:860-868.
Ho Y-C, Lee H-P, Hwang S-M, Lo W-H, Chen H-C, Chung C-K, Hu Y-C. 2006. Baculovirus transduction of human mesenchymal stem cell-derived progenitor cells: variation of transgene expression with cellular differentiation states. Gene Ther. 13:1471-1479.
Hofmann C, Sandig V, Jennings G, Rudolph M, Schlag P, Strauss M. 1995. Efficient gene-transfer into human hepatocytes by baculovirus vectors. Proc. Natl. Acad. Sci. U.S.A. 92(22):10099-10103.
Hu Y-C. 2006. Baculovirus vectors for gene therapy. Adv. Virus. Res. 68:287-320.
Hu Y-C. 2008. Baculoviral vectors for gene delivery: A review. Curr. Gene Ther. 8:54-65.
Huang AH, Stein A, Tuan RS, Mauck RL. 2009. Transient exposure to transforming growth factor beta 3 improves the mechanical properties of mesenchymal stem cell-laden cartilage constructs in a density-dependent manner. Tissue Eng Part A 15(11):3461-72.
Huang JI, Kazmi N, Durbhakula MM, Hering TM, Yoo JU, Johnstone B. 2005. Chondrogenic potential of progenitor cells derived from human bone marrow and adipose tissue: a patient-matched comparison. J Orthop Res 23(6):1383-9.
Im GI, Kim HJ, Lee JH. 2011. Chondrogenesis of adipose stem cells in a porous PLGA scaffold impregnated with plasmid DNA containing SOX trio (SOX-5,-6 and -9) genes. Biomaterials 32(19):4385-92.
Jin X, Sun Y, Zhang K, Wang J, Shi T, Ju X, Lou S. 2007a. Ectopic neocartilage formation from predifferentiated human adipose derived stem cells induced by adenoviral-mediated transfer of hTGF beta2. Biomaterials 28(19):2994-3003.
Jin XB, Sun YS, Zhang K, Wang J, Shi TP, Ju XD, Lou SQ. 2008. Tissue engineered cartilage from hTGF beta2 transduced human adipose derived stem cells seeded in PLGA/alginate compound in vitro and in vivo. J Biomed Mater Res A 86(4):1077-87.
Kemmis CM, Vahdati A, Weiss HE, Wagner DR. 2010. Bone morphogenetic protein 6 drives both osteogenesis and chondrogenesis in murine adipose-derived mesenchymal cells depending on culture conditions. Biochemical and Biophysical Research Communications 401(1):20-25.
Kinner B, Capito RM, Spector M. 2005. Regeneration of articular cartilage. Adv Biochem Eng Biotechnol 94:91-123.
Ko Y-G, Kawazoe N, Tateishi T, Chen G. 2010. Preparation of chitosan scaffolds with a hierarchical porous structure. Journal of Biomedical Materials Research Part B: Applied Biomaterials 9999B:NA-NA.
Kolostova K, Taltynov O, Pinterova D, Boubelik M, Raska O, Hozak P, Jirkovska M, Bobek V. 2011. Wound healing gene therapy: cartilage regeneration induced by vascular endothelial growth factor plasmid. Am J Otolaryngol.
Komsa-Penkova R, Spirova R, Bechev B. 1996. Modification of Lowry's method for collagen concentration measurement. J Biochem Biophys Methods 32(1):33-43.
Kost TA, Condreay JP. 2002. Recombinant baculoviruses as mammalian cell gene-delivery vectors. Trends Biotechnol 20(4):173-80.
Kuroda R, Usas A, Kubo S, Corsi K, Peng H, Rose T, Cummins J, Fu FH, Huard J. 2006. Cartilage repair using bone morphogenetic protein 4 and muscle-derived stem cells. Arthritis & Rheumatism 54(2):433-442.
Lee DK, Choi KB, Oh IS, Song SU, Hwang S, Lim CL, Hyun JP, Lee HY, Chi GF, Yi Y and others. 2005. Continuous transforming growth factor beta1 secretion by cell-mediated gene therapy maintains chondrocyte redifferentiation. Tissue Eng 11(1-2):310-8.
Lee JB, Kim SE, Heo DN, Kwon IK, Choi B-J. 2010. In vitro characterization of nanofibrous PLGA/gelatin/hydroxyapatite composite for bone tissue engineering. Macromolecular Research 18(12):1195-1202.
Lee KBL, Hui JHP, Song IC, Ardany L, Lee EH. 2007. Injectable Mesenchymal Stem Cell Therapy for Large Cartilage Defects-A Porcine Model. Stem Cells 25(11):2964-2971.
Leisy DJ, Lewis TD, Leong JA, Rohrmann GF. 2003. Transduction of cultured fish cells with recombinant baculoviruses. J Gen Virol 84(Pt 5):1173-8.
Leng P, Ding CR, Zhang HN, Wang YZ. 2012. Reconstruct large osteochondral defects of the knee with hIGF-1 gene enhanced Mosaicplasty. Knee 19(6):804-11.
Lewis PB, McCarty LP, 3rd, Kang RW, Cole BJ. 2006. Basic science and treatment options for articular cartilage injuries. J Orthop Sports Phys Ther 36(10):717-27.
Li X-k, Cai S-x, Liu B, Xu Z-l, Dai X-z, Ma K-w, Li S-q, Yang L, Sung KLP, Fu X-b. 2007. Characteristics of PLGA–gelatin complex as potential artificial nerve scaffold. Colloids and Surfaces B: Biointerfaces 57(2):198-203.
Liao J, Guo X, Grande-Allen KJ, Kasper FK, Mikos AG. 2010. Bioactive polymer/extracellular matrix scaffolds fabricated with a flow perfusion bioreactor for cartilage tissue engineering. Biomaterials 31(34):8911-20.
Lim SM, Oh SH, Lee HH, Yuk SH, Im GI, Lee JH. 2010. Dual growth factor-releasing nanoparticle/hydrogel system for cartilage tissue engineering. Journal of Materials Science: Materials in Medicine 21(9):2593-2600.
Lin CY, Lin KJ, Kao CY, Chen MC, Lo WH, Yen TC, Chang YH, Hu YC. 2011. The role of adipose-derived stem cells engineered with the persistently expressing hybrid baculovirus in the healing of massive bone defects. Biomaterials 32(27):6505-14.
Liu L, DiGirolamo CM, Navarro PA, Blasco MA, Keefe DL. 2004. Telomerase deficiency impairs differentiation of mesenchymal stem cells. Exp Cell Res 294(1):1-8.
Lu CH, Lin KJ, Chiu HY, Chen CY, Yen TC, Hwang SM, Chang YH, Hu YC. 2012. Improved Chondrogenesis and Engineered Cartilage Formation from TGF-beta3-Expressing Adipose-Derived Stem Cells Cultured in the Rotating-Shaft Bioreactor. Tissue Eng Part A.
Lu H, Ko Y-G, Kawazoe N, Chen G. 2010. Cartilage tissue engineering using funnel-like collagen sponges prepared with embossing ice particulate templates. Biomaterials 31(22):5825-5835.
Ma L, Tamarina N, Wang Y, Kuznetsov A, Patel N, Kending C, Hering BJ, Philipson LH. 2000. Baculovirus-mediated gene transfer into pancreatic islet cells. Diabetes 49(12):1986-91.
Mahmoudifar N, Doran PM. 2010. Chondrogenic differentiation of human adipose-derived stem cells in polyglycolic acid mesh scaffolds under dynamic culture conditions. Biomaterials 31(14):3858-67.
Marolt D, Augst A, Freed LE, Vepari C, Fajardo R, Patel N, Gray M, Farley M, Kaplan D, Vunjak-Novakovic G. 2006. Bone and cartilage tissue constructs grown using human bone marrow stromal cells, silk scaffolds and rotating bioreactors. Biomaterials 27(36):6138-6149.
Mazzetti I, Grigolo B, Pulsatelli L, Dolzani P, Silvestri T, Roseti L, Meliconi R, Facchini A. 2001. Differential roles of nitric oxide and oxygen radicals in chondrocytes affected by osteoarthritis and rheumatoid arthritis. Clin Sci (Lond) 101(6):593-9.
Mehlhorn AT, Niemeyer P, Kaschte K, Muller L, Finkenzeller G, Hartl D, Sudkamp NP, Schmal H. 2007. Differential effects of BMP-2 and TGF-beta1 on chondrogenic differentiation of adipose derived stem cells. Cell Prolif 40(6):809-23.
Merceron C, Vinatier C, Portron S, Masson M, Amiaud J, Guigand L, Cherel Y, Weiss P, Guicheux J. 2010. Differential effects of hypoxia on osteochondrogenic potential of human adipose-derived stem cells. Am J Physiol Cell Physiol 298(2):C355-64.
Meyer EG, Buckley CT, Thorpe SD, Kelly DJ. 2010. Low oxygen tension is a more potent promoter of chondrogenic differentiation than dynamic compression. J Biomech 43(13):2516-23.
Miljkovic N, Cooper G, Marra K. 2008. Chondrogenesis, bone morphogenetic protein-4 and mesenchymal stem cells. Osteoarthritis and Cartilage 16(10):1121-1130.
Minehara H, Urabe K, Naruse K, Mehlhorn AT, Uchida K, Südkamp NP, Itoman M. 2010. A new technique for seeding chondrocytes onto solvent-preserved human meniscus using the chemokinetic effect of recombinant human bone morphogenetic protein-2. Cell and Tissue Banking.
Murrell GA, Doland MM, Jang D, Szabo C, Warren RF, Hannafin JA. 1996. Nitric oxide: an important articular free radical. J Bone Joint Surg Am 78(2):265-74.
Nakagawa S, Arai Y, Mazda O, Kishida T, Takahashi KA, Sakao K, Saito M, Honjo K, Imanishi J, Kubo T. 2010. N-acetylcysteine prevents nitric oxide-induced chondrocyte apoptosis and cartilage degeneration in an experimental model of osteoarthritis. J Orthop Res 28(2):156-63.
Newman AP. 1998. Articular cartilage repair. Am J Sports Med 26(2):309-24.
Obert L, Lepage D, Gindraux F, Garbuio P. 2009. Bone morphogenetic proteins in soft-tissue reconstruction. Injury 40 Suppl 3:S17-20.
Otsuki S, Hanson SR, Miyaki S, Grogan SP, Kinoshita M, Asahara H, Wong CH, Lotz MK. 2010. Extracellular sulfatases support cartilage homeostasis by regulating BMP and FGF signaling pathways. Proc Natl Acad Sci U S A 107(22):10202-7.
Pagnotto MR, Wang Z, Karpie JC, Ferretti M, Xiao X, Chu CR. 2007. Adeno-associated viral gene transfer of transforming growth factor-β1 to human mesenchymal stem cells improves cartilage repair. Gene Therapy 14(10):804-813.
Palmieri B, Lodi D, Capone S. 2010. Osteoarthritis and degenerative joint disease: local treatment options update. Acta Biomed 81(2):94-100.
Park J, Gelse K, Frank S, von der Mark K, Aigner T, Schneider H. 2006. Transgene-activated mesenchymal cells for articular cartilage repair: a comparison of primary bone marrow-, perichondrium/periosteum- and fat-derived cells. J Gene Med 8(1):112-25.
Pearle AD, Warren RF, Rodeo SA. 2005. Basic science of articular cartilage and osteoarthritis. Clin Sports Med 24(1):1-12.
Peng H, Zhou JL, Liu SQ, Hu QJ, Ming JH, Qiu B. 2010. Hyaluronic acid inhibits nitric oxide-induced apoptosis and dedifferentiation of articular chondrocytes in vitro. Inflamm Res 59(7):519-30.
Perron JK, Naguib HE, Daka J, Chawla A, Wilkins R. 2009. A study on the effect of degradation media on the physical and mechanical properties of porous PLGA 85/15 scaffolds. J Biomed Mater Res B Appl Biomater 91(2):876-86.
Petrenko YA, Ivanov RV, Petrenko AY, Lozinsky VI. 2011. Coupling of gelatin to inner surfaces of pore walls in spongy alginate-based scaffolds facilitates the adhesion, growth and differentiation of human bone marrow mesenchymal stromal cells. J Mater Sci Mater Med 22(6):1529-40.
Pilgaard L, Lund P, Duroux M, Lockstone H, Taylor J, Emmersen J, Fink T, Ragoussis J, Zachar V. 2009. Transcriptional signature of human adipose tissue-derived stem cells (hASCs) preconditioned for chondrogenesis in hypoxic conditions. Experimental Cell Research 315(11):1937-1952.
Ponnazhagan S, Mukherjee P, Wang XS, Qing K, Kube DM, Mah C, Kurpad C, Yoder MC, Srour EF, Srivastava A. 1997. Adeno-associated virus type 2-mediated transduction in primary human bone marrow-derived CD34+ hematopoietic progenitor cells: donor variation and correlation of transgene expression with cellular differentiation. J Virol 71(11):8262-7.
Puetzer JL, Petitte JN, Loboa EG. 2010. Comparative review of growth factors for induction of three-dimensional in vitro chondrogenesis in human mesenchymal stem cells isolated from bone marrow and adipose tissue. Tissue Eng Part B Rev 16(4):435-44.
Saini S, Wick TM. 2003. Concentric cylinder bioreactor for production of tissue engineered cartilage: effect of seeding density and hydrodynamic loading on construct development. Biotechnol Prog 19(2):510-21.
Saldanha KJ, Doan RP, Ainslie KM, Desai TA, Majumdar S. 2011. Micrometer-sized iron oxide particle labeling of mesenchymal stem cells for magnetic resonance imaging-based monitoring of cartilage tissue engineering. Magn Reson Imaging 29(1):40-9.
Santoro R, Olivares AL, Brans G, Wirz D, Longinotti C, Lacroix D, Martin I, Wendt D. 2010. Bioreactor based engineering of large-scale human cartilage grafts for joint resurfacing. Biomaterials 31(34):8946-52.
Sarkis C, Serguera C, Petres S, Buchet D, Ridet JL, Edelman L, Mallet J. 2000. Efficient transduction of neural cells in vitro and in vivo by a baculovirus-derived vector. Proc Natl Acad Sci U S A 97(26):14638-43.
Schulz RM, Bader A. 2007. Cartilage tissue engineering and bioreactor systems for the cultivation and stimulation of chondrocytes. Eur Biophys J 36(4-5):539-68.
Senta H, Park H, Bergeron E, Drevelle O, Fong D, Leblanc E, Cabana F, Roux S, Grenier G, Faucheux N. 2009. Cell responses to bone morphogenetic proteins and peptides derived from them: Biomedical applications and limitations. Cytokine & Growth Factor Reviews 20(3):213-222.
Shen B, Wei A, Tao H, Diwan AD, Ma DD. 2009. BMP-2 enhances TGF-beta3-mediated chondrogenic differentiation of human bone marrow multipotent mesenchymal stromal cells in alginate bead culture. Tissue Eng Part A 15(6):1311-20.
Smith GE, Summers MD, Fraser MJ. 1992. Production of human beta interferon in insect cells infected with a baculovirus expression vector. 1983. Biotechnology 24:434-43.
Sommar P, Pettersson S, Ness C, Johnson H, Kratz G, Junker JPE. 2010. Engineering three-dimensional cartilage- and bone-like tissues using human dermal fibroblasts and macroporous gelatine microcarriers. Journal of Plastic, Reconstructive & Aesthetic Surgery 63(6):1036-1046.
Steadman JR, Rodkey WG, Rodrigo JJ. 2001. Microfracture: surgical technique and rehabilitation to treat chondral defects. Clin Orthop Relat Res(391 Suppl):S362-9.
Strem BM, Hicok KC, Zhu M, Wulur I, Alfonso Z, Schreiber RE, Fraser JK, Hedrick MH. 2005. Multipotential differentiation of adipose tissue-derived stem cells. Keio J Med 54(3):132-41.
Sung L-Y, Lo W-H, Chiu H-Y, Chen H-C, Chuang C-K, Lee H-P, Hu Y-C. 2007. Modulation of chondrocyte phenotype via baculovirus-mediated growth factor expression. Biomaterials 28:3437-3447.
Taipaleenmaki H, Suomi S, Hentunen T, Laitalaleinonen T, Saamanen A. 2008. Impact of stromal cell composition on BMP-induced chondrogenic differentiation of mouse bone marrow derived mesenchymal cells. Experimental Cell Research 314(13):2400-2410.
Tan H, Huang D, Lao L, Gao C. 2009. RGD modified PLGA/gelatin microspheres as microcarriers for chondrocyte delivery. Journal of Biomedical Materials Research Part B: Applied Biomaterials 91B(1):228-238.
Tanaka Y, Yamaoka H, Nishizawa S, Nagata S, Ogasawara T, Asawa Y, Fujihara Y, Takato T, Hoshi K. 2010. The optimization of porous polymeric scaffolds for chondrocyte/atelocollagen based tissue-engineered cartilage. Biomaterials 31(16):4506-16.
Tay LX, Ahmad RE, Dashtdar H, Tay KW, Masjuddin T, Ab-Rahim S, Chong PP, Selvaratnam L, Kamarul T. 2012. Treatment outcomes of alginate-embedded allogenic mesenchymal stem cells versus autologous chondrocytes for the repair of focal articular cartilage defects in a rabbit model. Am J Sports Med 40(1):83-90.
Tetlow LC, Adlam DJ, Woolley DE. 2001. Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage: associations with degenerative changes. Arthritis Rheum 44(3):585-94.
Thorpe SD, Buckley CT, Vinardell T, O'Brien FJ, Campbell VA, Kelly DJ. 2010. The response of bone marrow-derived mesenchymal stem cells to dynamic compression following TGF-beta3 induced chondrogenic differentiation. Ann Biomed Eng 38(9):2896-909.
Tigli RS, Gumusderelioglu M. 2009. Chondrogenesis on BMP-6 loaded chitosan scaffolds in stationary and dynamic cultures. Biotechnol Bioeng 104(3):601-10.
Toh WS, Guo XM, Choo AB, Lu K, Lee EH, Cao T. 2009. Differentiation and enrichment of expandable chondrogenic cells from human embryonic stem cells in vitro. J Cell Mol Med 13(9B):3570-90.
Toh WS, Lee EH, Guo X-M, Chan JKY, Yeow CH, Choo AB, Cao T. 2010. Cartilage repair using hyaluronan hydrogel-encapsulated human embryonic stem cell-derived chondrogenic cells. Biomaterials 31(27):6968-6980.
Uitto J, Hoffmann HP, Prockop DJ. 1977. Purification and partial characterization of the type II procollagen synthesized by embryonic cartilage cells. Arch Biochem Biophys 179(2):654-62.
Vidal MA, Robinson SO, Lopez MJ, Paulsen DB, Borkhsenious O, Johnson JR, Moore RM, Gimble JM. 2008. Comparison of chondrogenic potential in equine mesenchymal stromal cells derived from adipose tissue and bone marrow. Vet Surg 37(8):713-24.
Vukicevic S, Grgurevic L. 2009. BMP-6 and mesenchymal stem cell differentiation. Cytokine & Growth Factor Reviews 20(5-6):441-448.
Wang QG, Magnay JL, Nguyen B, Thomas CR, Zhang Z, El Haj AJ, Kuiper NJ. 2009. Gene expression profiles of dynamically compressed single chondrocytes and chondrons. Biochem Biophys Res Commun 379(3):738-42.
Wei Y, Hu Y, Hao W, Han Y, Meng G, Zhang D, Wu Z, Wang H. 2008. A novel injectable scaffold for cartilage tissue engineering using adipose-derived adult stem cells. Journal of Orthopaedic Research 26(1):27-33.
Wei Y, Hu Y, Lv R, Li D. 2006. Regulation of adipose-derived adult stem cells differentiating into chondrocytes with the use of rhBMP-2. Cytotherapy 8(6):570-579.
Weinstein RS. 2012. Glucocorticoid-induced osteoporosis and osteonecrosis. Endocrinol Metab Clin North Am 41(3):595-611.
Wickham MQ, Erickson GR, Gimble JM, Vail TP, Guilak F. 2003. Multipotent stromal cells derived from the infrapatellar fat pad of the knee. Clin Orthop Relat Res(412):196-212.
Williams CG, Kim TK, Taboas A, Malik A, Manson P, Elisseeff J. 2003. In vitro chondrogenesis of bone marrow-derived mesenchymal stem cells in a photopolymerizing hydrogel. Tissue Eng 9(4):679-88.
Williams GM, Chan EF, Temple-Wong MM, Bae WC, Masuda K, Bugbee WD, Sah RL. 2010. Shape, loading, and motion in the bioengineering design, fabrication, and testing of personalized synovial joints. J Biomech 43(1):156-65.
Williamson AK, Chen AC, Sah RL. 2001. Compressive properties and function-composition relationships of developing bovine articular cartilage. J Orthop Res 19(6):1113-21.
Wu S-C, Chang J-K, Wang C-K, Wang G-J, Ho M-L. 2010. Enhancement of chondrogenesis of human adipose derived stem cells in a hyaluronan-enriched microenvironment. Biomaterials 31(4):631-640.
Xue D, Zheng Q, Zong C, Li Q, Li H, Qian S, Zhang B, Yu L, Pan Z. 2010. Osteochondral repair using porous poly(lactide-co-glycolide)/nano-hydroxyapatite hybrid scaffolds with undifferentiated mesenchymal stem cells in a rat model. Journal of Biomedical Materials Research Part A 94A(1):259-270.
Yanez R, Lamana ML, Garcia-Castro J, Colmenero I, Ramirez M, Bueren JA. 2006. Adipose tissue-derived mesenchymal stem cells have in vivo immunosuppressive properties applicable for the control of the graft-versus-host disease. Stem Cells 24(11):2582-91.
Yang B, Yin Z, Cao J, Shi Z, Zhang Z, Song H, Liu F, Caterson B. 2010. In vitrocartilage tissue engineering using cancellous bone matrix gelatin as a biodegradable scaffold. Biomedical Materials 5(4):045003.
Yao Y, Zhang F, Zhou R, Su K, Fan J, Wang D-A. 2010. Effects of combinational adenoviral vector-mediated TGFβ3 transgene and shRNA silencing type I collagen on articular chondrogenesis of synovium-derived mesenchymal stem cells. Biotechnology and Bioengineering 106(5):818-828.
Ye C, Hu P, Ma MX, Xiang Y, Liu RG, Shang XW. 2009. PHB/PHBHHx scaffolds and human adipose-derived stem cells for cartilage tissue engineering. Biomaterials 30(26):4401-6.
Zhu Y, Liu T, Song K, Fan X, Ma X, Cui Z. 2008. Adipose-derived stem cell: a better stem cell than BMSC. Cell Biochem Funct 26(6):664-75.
Zuk PA, Zhu M, Mizuno H, Huang J, Futrell JW, Katz AJ, Benhaim P, Lorenz HP, Hedrick MH. 2001. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng 7(2):211-28.