van Eijk Floor, Saris Daniel B F, Fedorovich Natalja E, Kruyt Moyo C, Willems W Jaap, Verbout Abraham J, Martens Anton C, Dhert Wouter J A, Creemers Laura
Department of Orthopaedics, University Medical Center Utrecht, Utrecht, The Netherlands.
Tissue Eng Part A. 2009 Oct;15(10):3109-17. doi: 10.1089/ten.TEA.2008.0541.
Ligament tissue engineering based on cell-seeded biomechanically functional constructs is a commonly studied strategy toward native anterior cruciate ligament replacement. Little is known about the survival and differentiation of the seeded cells after the transplantation. We applied retroviral genetic marking to trace implanted cells and studied their differentiation by species-specific immunolabeling of the extracellular matrix produced. Goat bone marrow stromal cells were transduced with a MoMuLV-based vector encoding the DeltaLNGFR gene. Transduced cells were seeded onto poly(lactic-co-glycolic acid) (PLGA) fibers and implanted subcutaneously into nude mice and left for various periods up to 6 weeks. Immunohistochemistry for LNGFR expression showed survival of the seeded cells after transplantation for up to 6 weeks. Immunohistochemistry for collagen type I and III showed the production of fibrous tissue inside the scaffolds. Moreover, using a goat-specific anti-collagen type III, donor-derived matrix could be demonstrated. We conclude that bone marrow stromal cells survived in vivo and at least partially differentiated after implantation.
基于接种细胞的生物力学功能构建体的韧带组织工程是一种常用于天然前交叉韧带置换的研究策略。关于移植后接种细胞的存活和分化情况知之甚少。我们应用逆转录病毒基因标记来追踪植入细胞,并通过对所产生的细胞外基质进行物种特异性免疫标记来研究它们的分化。用编码DeltaLNGFR基因的基于MoMuLV的载体转导山羊骨髓基质细胞。将转导的细胞接种到聚乳酸-乙醇酸共聚物(PLGA)纤维上,皮下植入裸鼠体内,并放置不同时间段直至6周。对LNGFR表达进行免疫组织化学检测显示,移植后接种细胞存活长达6周。对I型和III型胶原进行免疫组织化学检测显示,支架内有纤维组织产生。此外,使用山羊特异性抗III型胶原,可以证明有供体来源的基质。我们得出结论,骨髓基质细胞在体内存活,植入后至少部分发生了分化。